Genetics

GENETICS
Fundamentals and
Applications
Dr. H. C. Srivastava
Dr. Debmalya Barh
International Book Distributing Co.

GENETICS
Fundamentals and
Applications
Dr. H C Srivastava
M.Sc.(Agri.),Ph.D.(Genetics),
German Dip., Extension Cert.
Former Principal Scientist & Head (Medicinal & Aromatic Crops),
Indian Institute of Horticultural Research,
Bangalore-560089, India
Dr. Debmalya Barh

M.5c. (Agri-Horticulture), M.Se (Applied Genetics),
M.Tech and M.Phill in Biotechnology,
Ph.D. (Genomics), PGD in Molecular Biology (uSe, Bangalore),
PGD in Bioinformatics, PGDM (ABS), PGDIPRL, DST
Principal Scientist
Centre for Genomics and Applied Gene Technology
P.O. Nonakuri, Distt. Purva Medinipur-721172, West Bengal, India
International Book Distributing Co.

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Preface
Genetics is an integrated domain of any advanced field of biology in
2pt century. However, successful development of future applications
will depend on the basic domain knowledge. Genetics: Fundamentals
and Applications covers maximum aspects of genetics and it is aimed
at readers from multidisciplinary fields of biology.
It is an effort to use simple language and examples throughout the
book for better understanding of principle of genetics and its
application in modern era. Few specific chapters have been
incorporated to emphasize the application aspects of genetics in
disease pathology and therapeutic areas.
Basic principles of genetics (first 19 chapters) have been described in
a cognized way with simplest example sets for beginners at under
graduate and post graduate level. In contrary, Application part (last
5 chapters) especially chapters covering cancer genetics, apoptosis
and stem cell are written elaborately for post graduate students and
researchers in biomedical sciences.
Topics and Features
• Consists total 25 chapters covering maximum aspects in genetics.
• Simple understanding of genetic principles in plants and animals.
• Simple example for complex genetic problems.
• Includes a chapter for different terminologies used in genetics.
• Includes a chapter from agricultural point of view.
• Includes a chapter for cancer biology, genetics, diagnosis and
treatments.
• Explains different process and gene families in cancer.
• Highlights the challenges in conventional cancer chemotherapy.
• Includes a chapter for various aspects of apoptotic including
pathways and gene families in model organisms.
• Highlights cancer chemoprevention through apoptosis by dietary
phytochemicals.
• Includes techniques in detection of apoptosis.
• Describes in death biology, markers, plasticity, trans-
differentiation and therapeutic applications of different stem
cells.
• 64 illustrations for easily understanding of the subject matter.
Dr. H C Srivastava
Dr. Debmalya Barh
"This page is Intentionally Left Blank"
ACKNOWLEDGEMENTS AND DEDICATIONS
The authors express thanks to all the scientists whose work has been
referred in this book. Dr. Srivastava is highly grateful to his wife
Mrs. Ratan Rani for her support & encouragement. Thanks are also
to Mr. Neeraj, Mrs. Monika, Mr. Pankaj, Dr. Priti, Mr. Paresh,
Mrs. Meghna, Mrs. Sarla & Mrs. Sarojni for their assistance.
Dr. Srivastava dedicates this book to his Late Grand Father & Grand
Mother Mr. & Mrs. Matabadal Lal Srivastava; Late Father & Mother
Mr. Daya Shankar Lal Srivastava, Mrs. Indrakali Devi Srivastava; Late
Uncle Mr. Badri Prasad Srivastava and Late Younger Brother Dr.
Sushil Kumar Srivastava.
Dr. Debmalya dedicates the work to his loving parents Mr. Purnendu
and Mrs. Mamata.
/'
iii
PART 1- Fundamentals of Genetics
Contents
Chapter Topic Page No.
No.
1 Introduction 3
2 Brief History 7
3 Important Definitions of Genetics 13
4 Chromosome and Its Structure 23
5 Mitosis and Meiosis 28
6 Laws of Inheritance 37
7 Heterosis or Hybrid Vigor 67
8 Mutation 71
9 Polyploidy 79
10 Linkage 87
11 Crossing Over 94
12 Chromosome Map 100
13 Chromosomal Aberrations 107
14 Cytoplasmic Inheritance 112
15 Sex Determination 115
16 Sex Linked Inheritance 125
17 The Gene 131
18 The Gene Complex 141
19 Gene Action 144
List of Figures
Fig. No. Descri~tion Page No.
1 Structure of a typical chromosome. 23
2 Shape of chromosome depending on centromeric 25
position.
3 Salivary gland chromosomes of fruit fly. 26
4 Various stage of mitosis. 29
5 Various stages of meiosis. 34 '"
6 Mortohybrid cross between pea flowers. 40

7 Monohybrid cross between white and red cows. 40
8 Incomplete dominance in 4 O'clock plant. 41 ~
~
9 Hybridization between pink and white flower pea. 42
10 Process of assortment of chromatids during 44
gamete formation.
11
12
13
14
Dihybrid cross.
Dihybrid cross.
Testcross.
Backcross.
44
46
52
53
1
,
iv
1
Fig. No. Descri~tion Page No.
15 Hybrid vigour in maize. 67
16 CIB method of detecting lethal mutation. 77
17 Origin of auto and allopolyploids. 80
18 Raphno brassica. 80
19 Recombination of genes. 89
20 Mechanism of crossing over. 95
21 Cytological proof of crossing over. 96
22 Contact theory of Crossing over. 98
23 Breakage theory of Crossing over. 98
24 Crossing over between A and C or double cross 104
over.
25 Chromosome map of Drosophila melanogaster. 105
26 Different types of deletions. 108
27 Different types of translocations. 109
28 Types of inversion and inversion loop. 110
29 Sex determination systems. 118
30 Genic balance theory. 120
31 Effect on sex of the balance between X 122
chromosomes and autosomes.
32 Intersex in Drosophila melanogaster. 122
33 Gynandromorph of Drosophila melanogaster. 123
34 Structure of human sex chromosomes. 126
35 Inheritance of color blindness. 129
36 Inheritance of sex linked character through Y 130
chromosome.
37 Double helical structure of DNA (gene). 133
38 Chemical structure of ribose and deoxyribose. 134
39 Structure of purine and pyrimidine base. 135
40 DNA double hehix. 136
41 Action of ultraviolet rays on gene in producing 138
mutation.
42 Action of HNO 2 on gene in producing mutation. 139
43 Action of 5 Br-uracil on gene in producing 139
mutation.
44 Formation of pigment from Tryptophan. 148
45 Action of genes KK and kk in producing disease 151
phenylketonuria.
v
PART 11- Applied Genetics
Contents
Chapter Topic Page No.
No.
1 Cancer Genetics 155
2 Apoptosis 178
3 Stem Cell 187
4 Eugenics 215
5 Plant Breeding 222
List of Figures
Figure Description Page No.
No.
1 Sequence of Adenocarcinoma 155
2 Different stages of cancer 156
3 Pathway events in cancer 166
4 Differentiation Gene hypothesis 167
5 Process of metastasis 168
6 Differences between apoptosis and necrosis 179
7 General A poptotic pathways 181
8 Apoptosis in C. elegans 182
9 Apoptosis in Drosophila 182
10 Types of stem cell division 189
11 Potentiality of ESC 190
12 In vivo differentiation of ESC 196
13 In vitro differentiation of ESC 197
14 Trans-differentiation of adult stem cell 199
15 Cell based therapeutics 200
16 Strategy for stem cell therapy 203
17 In vitro differentiation of BMSC 205
18 Trans-differentiation of MSC 208
19 Approaches to Stem Cell Therapy for Cerebral 214
Infarction
Index 235
vi
(Part I
Fundamental of
Genetics
Chapter 1
Introduction
Genetics is an important branch of biology. It means study of heredity
and variation. Heredity relates to that science by which living beings
resembles to their relatives and ancestors. Variation means the science
by which living being differs from their relatives and ancestors. Thus
genetics is the ~cience to study how various characters are transferred
from parents and develop into offsprings. Now-a-days the sphere of
genetics is extremely wide. It studies how genes are developed, what
is their chemical structure and how these effect the development and
behaviour of living beings. Genetics has been divided into six
branches: 1. Cytogenetics, 2. Physiological genetics, 3 Biochemical
genetics, 4. Population genetics, 5. Eugenics and 6. Applied genetics
including cancer genetics, apoptosis & stem cell.
Importance of genetics:
Genetics is comparatively a recent science. Its development has
happened at fast speed. Genetics has been widely used to develop
improved types of cereals, vegetables, fruits, ornamental plants,
medicinal & aromatic plants, spices etc. and various animals. In
addition to this, genetics is closely associated with other sciences such
as morphology, taxonomy, embryology, cytology, biochemistry,
ecology, evolution, bacteriology, statistics etc., as briefed below:-
1. Genetics and Morphology: Morphologies studies various
morphological features of living beings. Growth and
development of tissues are effected by mutations - a branch of
genetics
2. Genetics and Taxonomy: Classification of various living beings,
their species, genus, family and order depends on their genetics.
Development of a living being is governed by its genetical
constitution.
3. Genetics and Embryology: Theoretical and practical embryology
is governed by its genetics. Genetical constitution of a living
being conditions its development.
3
Genetics: Fundamentals and Applications
4. Genetics and Cytology: Cytology of a living being is concerned

with its genetics. Studies on Cytology and genetics are collectively
grouped as cytogenetics. Dunn (1960) has rightly mentioned
that cytology is the pillar of genetics.
5. Genetics and Biochemistry: To understand the nature and
function of genetic material a geneticist has to take help of
biochemistry. Nowadays genetics helps in biochemical research.
For example due to reduction of an amino acid tyrosine by an
enzyme, homo genes tic acid is formed in human body which
causes a disease Alkatopnuria in which urin on keeping in air
turns black. This disease is controlled by a simple genetic factor.
Another example of controlling enzyme by genetic factor has
been found in the fungus Neurospora in which formation of
spores gets stopped.
6. Genetics and Ecology: Ecology means study of living being in
natural environment. Now a day's Significant progress has been
made in ecology by the help of genetics. Acclamatization of
various polyploids in different environment reveals close
relationship between ecology and genetics. This fact reveals that
ecology is dependent on heredity of the organism.
7. Genetics and Psychology: In modem days genetics is used to
solve psychological problems. Studies of inheritance and DNA
help to solve several psychological problems.
8. Genetics and Statistics: Biometry helps significantly in genetical
research. Dunn (1960) has rightly mentioned that statistics has
greatly helped in development of genetics.
9. Genetics and Bacteriology: Several scientists have done
remarkable progress on physiology, pathology, biochemistry etc.,
of bacteria. But complete information on their life cycle could
not be obtained. It was afterwards completed by geneticist, the
invisible cytological facts were proved by practical experiments.
For example sex linked inheritance is expressed in presence of
sex chromosomes, though cytologically it mayor may not be
visible. Similarly, genetics has been helpful in studies of virus.
10. Genetics and Botany & Zoology: Botany and Zoology are
different sciences for studies of plants and animals. However,
genetics correlates both these sciences. Both express similarly in
inheritance.
4
Introduction
Application of Genetics:
Genetics has been used for varietal improvement of various plants &
breed amelioration of various animals, for treatment of diseases, and
for betterment of human race. Brief descriptions of various
applications are mentioned below:
1. Improvement of agricultural plants:
a. Improvement in disease resistance: One of the main agricultural
problems to the whole world is occurrence of diseases which
cause economic loss to crops. The best solution to this problem
is development of disease resistant varieties using the principles
of genetics.
b. Amelioration of insect resistance: Crops also suffers serious
economic loss due to various types of insects such as aphids,
sucking insects, mites, nematodes etc. This problem can also be
solved by breeding for resistance following the princip1es of
genetics.
c. Synthesizing of plants with multiple qualities: With the help of
genetics nowadays plant breeder are synthesizing plants with
multiple qualities such as high yield, superior quality, resistance
to diseases & insects and more adoptability to the existing agro-
climate. Breeders do it by multiple crossing.
Details are discussed in the chapter on plant breeding. Plant breeding
is a subject which is related to genetics, cytology and cytogenetics
and has its own special features.
2. Use of genetics for improvement in animals:
a. Improvement for disease resistance: Due to occurrence of some
diseases in animals the farmer suffers economic loss. Several
medicines are invented to control these diseases. A better
method of control is to develop disease resistance breeds. For
example, typhoid resistant breeds of poultry.
b. Improvement in utility of domesticated animals: Such
improvement in animals can be possible by research on
quantitative inheritance & by help of proper hybridization and
selection.
3. Genetics for treatment of diseases:
Treatment of cancer is done by gene therapy, surgery, radiation
therapy, chemotherapy, hormone therapy, immunotherapy /
5
biological therapy, adjuvant therapy - combining two or more

treatments, prophylactic or preventive treatment. Diseases and
maladies treated using adult stem cells are brain cancer, ovarian cancer,
skin cancer, testicular cancer, tumors abdominal organs, chronic
myelogenous leukemia, acute lymphoblastic leukemia, breast cancer,
cancer of the lymph nodes, soft tissue sarcoma, various solid tumors
etc.
4. Genetics for betterment of human race:
This subject was named as Eugenics by Galton in 1904. In this science
geneticist investigate those factors which effect hereditary characters
in human and which can make such characters more useful to mankind.
Genetics can help to predict the future life of an infant. To improve
human being two methods may be used: first method is euthenics. It
means by improving the environment and surroundings. The second
method is eugenics. Under it improvement in hereditary constitution
are studied. This method is of two types- positive eugenics and
negative eugenics ie. to get rid of undesirable germplasms Details
are given in chapter on eugenics.
Hence it is apparent that progress of genetics has been associated
with progress of other sciences. Beyond academic importance,
genetics has tremendous applications for human welfare.
6
Chapter 2
Brief History
Centuries before Christian era, inEgypt and Mesopotamia few people
had some idea of genetics because tliey attempted cross breeding in
animals and plants. Example of artificial pollination in date palm was
available in history of that time. But more development in this science
started from beginning of twentieth century.
In eighteenth and nineteenth century geneticist were trying to
understand that how characters are transferred from one generation
to another. But actual knowledge of heredity did not come till details
of reproductive process were not clear. In 1760 Kolreuter a botanist
in Germany, on hybridization of one type of tobacco with other type
of tobacco (by transferring of pollen from one type to pistil of other
type) has observed that in the hybrid the characters came from both
the parents. From this observation it was clear that characters are
transferred from gametes of the parents. Therefore, gametes had
been said as a unit of heredity.
Based on these facts the Austrian botanist John Gregor Mendel has
experimented with sweet pea in his garden and found unprecedented
success. Earlier workers in this experiment failed. The reason for his
success was that at one time he selected one contrasting character.
He counted the offsprings of various types and kept the account
properly. First he experimented with plants having green & yellow
seeds and wrinkled & smooth seed coat. From these experiments he
concluded that inheritance is based on certain principles. He has
expressed that if pedigree of two parents is correctly known then it
will be possible to forecast the type and number of progenies. In the
end he concluded that inheritance is controlled by units or factors
present in the cell. He has also expressed that in somatic cells of
plants and animals there are two units or factors present but in their
sex cells i.e., pollen and ovule or sperm and egg cells only one unit or
factor is present. Mendel was unable to find out that how two units
or factors in somatic cells become only one unit in their sex cells. This
mystery was solved by the experiments of Fleming (1882). He has
told that it happens by the cell division mitosis and meiosis. Oscar
7
Genetics: FUlIdamelltals and Applications
Hertwig (1885) has told that in combining vf sex cells it is not that
two cells unite but the nucleus of male sex cell unites with the nucleus
of female sex cell. Therefore, he concluded that the uhit of inheritance
is nucleus. Immediately after knowledge of meiosis and fertilization
Devries from Holland, Correns from Germany and Von Tschermark
from Austria experimented separately on the laws of inheritance
proposed by John Gregor Mendel. These scientists have designated
the laws as Men del' s laws of inheritance.
Despite these information it could not be explained that how so many
characters get transferred from one generation to another. To solve
this mystery efforts were made by the famous scientist Charls Darwin
in 1895. Later on he proposed the theory of Pangenesis. It suggested
that every cell of the body produces its agents known as Gemmules.
These gemmules come in the blood stream. Then they reach in the
sex cells (ovaries and sperms). These sex cells unite and ultimately
develop a living being. Each gemmule helps in expression of different
character. But this theory was proved to be wrong by Galton (1822-
1911) and other scientists.
At almost same time the famous scientist Lamark (1844-1929) did
research on same problem of heredity. He experienced that a living
being or any of its organ gets changed due to use or no use or
environmental effect. For example if any organ is used constantly it
becomes strong, but if not used it becomes weak. Lamark thus
proposed a theory of acquired characters". He told that acquired
It
characters of parents reach to their offsprings. This theory became

famous as 'Lamarkism".
Weismann (1835-1914) could not accept Lamarkism. After
experimentation he concluded that reproductive cells are different
from other cells. So reproductive cells can be called as 'germ plasm'.
He told that germ plasm can give rise to somatoplasm. No changes in
the somatoplasm can go in germplasm. Therefore, acquired characters
cannot be inherited. Weismann has also proved it wrong by simple
experiments. He has cut tail of rats for several generations. But still
the new generation have normal tail. In muslims skin over the tip of
penis is removed from every child but still the newly born muslim
children have skin over tip of their penis. Weismann (1835-1914) has
proposed Germplasm Theory. It has significantly helped in
development of genetics.
Significant progress of genetics has started in 1875 when Strasburger
has invented that at cell division nucleus is changed to several
8
Brief History
longitudinal structures to which he has given the name chromosome.

The chromosomes are understood as the base of heredity. In 1902
Sutton has discovered that each chromosome is made up of globular
structures which are known as genes. Genes are arranged
longitudinally and control all the characters or traits. Afterwards,
especially after 1920 genetics progressed at tremendous speed.
Cancer is multistage carcinogenesis processes during which normal
dividing cells accumulate multiple somatic mutations i.e. point
mutations and mutations due to chromosomal damage, and then
become cancer cells by several pathway events. Most human tumors
are spontaneous but some are age and sex related, and a few are
inherited or familial. There are many etiologic factors such as genetic
history, diet, lifestyle, and environment may contribute to the
development of these tumors. In human latency period ranges from a
few years to 30 years or more.
Apoptosis is the term coined by Andrew Wyllie in 1972 to describe
morphological description of a dying cell that contrast with necrosis.
Programmed cell death (PCD) is a term used to descIibe cells that die
at predictable time and place during development. Programmed cell
death is a gene directed cellular suicide mechanism that eliminates
unwanted, superfluous cells during development as well as tissue
homeostasis, control proliferation and differentiation and defends
against viral infection. All Programmed cell deaths are apoptotic and
so these terms are used interchangeably. Programmed cell death is
associated with ALPS- atoimmunity, myoicordial infraction, stroke,
diabetes, neurodegenerative diseases, alzheimer's disease, infertility,
sepsis, viral infection etc.
Characters of apoptosis are cytoplasm shrinks with out membrane
rupture, blabbing of plasma membrane and nuclear membrane
chromatin condenses and migrates to nuclear membrane, inter-
chromosomal cleavage in nucleosomal size by Mg+2 dependent
endonuclease leads to laddering of DNA at nucleosomal repeat length
of 200bp, fragmentation of cell with retention of cell membrane,
flipping up phosphatidal serine from outer plasma membrane, cell
contents are packed in membrane bounded bodies (apoptotic bodies),
internal organneles still remain functional, mitocondria swells and
engulfed by neighbours, epitopes appear on plasma membrane,
making cell as phagocytotic target and no spillage and inflammation.
9
There are 3 different mechanisms of apoptosis: I-death signals

generated within the cell. 2-activation of death activators by extrinsic
death signals. 3-by reactive oxygen species and DNA damage.
Cancer is a multistage carcinogenic process where there is a net
accumulation of atypical cells arising from excess proliferation, an
insufficient apoptosis or a combination of the two. There are also
various side effects of chemotherapy. Thus from therapeutic point of
view, the best strategy is induced apoptosis in the neoplastic cell
11
line without affecting the normal cells of the body. Several diatary
phytochemicals are nowadays used for chemoprevention.
Stem cells differ from other kinds of cells in the body. All stem cells-
regardless of their source - have three general properties: I-they are
capable of dividing and renewing themselves for long periods; 2-
they are unspecialized; 3- regulated by intrinsic signals and the external
microenvironment and can give rise to specialized cell types.
Ability of stem cells is self-regeneration, to divide and produce more
stem cells. During early development, the cell division is symmetrical
i.e. each cell divides to gives rise to daughter cells each with the same
potential. Later in development, the cell divides asymmetrically with
one of the daughter cells produced also a stem cell and the other a
more differentiated cell.
Stem cells are involved in three processes - development, repair of
adult tissue and cancer. An adult stem cell is an undifferentiated cell
found among differentiated cells in a tissue or organ, can renew itself,
and can differentiate to yield the major specialized cell types of the
tissue or organ. The primary roles of adult stem cells in a living
organism are to maintain and repair the tissue in which they are found.
Some scientists now use the term somatic stem cell instead of adult
stem cell. Unlike embryonic stem cells cells, which are defined by
their origin, the origin of adult stem 'cells in mature tissues is unknown.
Bone marrow (BM) contains hematopoietic stem cells, which
differentiate into every type of mature blood cell; endothelial cell
progenitors; and marrow stromal cells, also called mesenchymal stem
cells, which can differentiate into mature cells of multiple mesenchymal
tissues including fat, bone, and cartilage. Recent findings indicate that
adult BM also contains cells that can differentiate into additional
mature, nonhematopoietic cells of multiple tissues including epithelial
cells of the liver, kidney, lung, skin, gastrointestinal (GI) tract, and
myocytes of heart and skeletal muscle. Experimental results obtained
10
Brief History
in vitro and in vivo are the subject of this review. The emphasis is on
how these experiments were performed and under what conditions
differentiation from bone marrow to epithelial and neural cells occurs.
Questions arise regarding whether tissue injury is necessary for this
differentiation and the mechanisms by which it occurs. We also
consider which bone marr-ow subpopulations are capable of this
differentiation. Only after we have a better understanding of the
mechanisms involved and of the cells required for this differentiation
will we be able to fully harness adult stem cell plasticity for clinical
purposes
Eugenics word was evolved by Sir Francis Galton in the year 1883.
He defined it as - eugenics is the study of those factors by which
development of mental and physical characters takes place in human
beings. In other words we can say that the branch of genetics in which
the laws of inheritance are used to improve human beings is called
eugenics. Eugenics is a Greek word meaning well born. Aims and
ideals of eugenics are improvementfor: 1- mental and physical healths,
2-intelligence, 3-moral character and 4- to attain specialization in
knowledge is very useful to the person and society. Generally the
capacity for specialization is hereditary.
Plant breeding is based on principles of genetics and studies in details
the technologies of crop improvement. A good plant breeder has good
understanding of genetics and cytogenetics. In addition to these he/
she should know agronomy, horticulture, plant pathology, plant
physiology, statistics, agricultural chemistry and entomology. Plant
breeding has been defined by G.M. Poehlman and D.N. Borthakur
(1959) as a science to improve heredity of plants. When by
hybridization between close relatives offspring are produced, it is
known as inbreeding. But if the parents are least related or distantly
related then the breeding is known as out breeding.
Under plant breeding stigma of other plant is artificially pollinated
and offspring are produced for various types of selections. Therefore
plant breeder has to know merits and demerits of the plant and
requirement of the country.
Objective of plant breeding is to combine various useful characters
into one variety of plant. The useful characters which a plant breeder
must understand are:high yield, early maturity, high quality. It should
have sufficient nutrient, attractive colour, should be tasty and easily
digestible, non shedding of grain, resistance to drought and frost,
disease and insect resistance, more medicinal property. In case of
11
Genetics: Fundamentals and ApplicatiollS
breeding of medicinal & aromatic crops the objective is to increase

yield & medicinal properties, more yield of superior quality essential
oil and resistance to biotic & abiotic stress. In addition to above
mentioned objectives a plant breeder makes efforts to develop
varieties which by application of more fertilizers and manure can
produce more yield, the fruits should have uniformly good size and
colour. The crop should have resistance to lodging and flood.
Main methods of plant breeding are plant introduction, selection,
hybridization, mutation and polyploidy. Hybridization in self
pollinated crops are pedigree selection method, bulk method, back
cross method, and multiple cross method. Hybridization procedures
in cross pollinated crops are I-single cross method 2- double cross
method. 3-triple cross method. Depending on parental relationships
hybridization can be of 3 types: intervarietal cross, interspecefic cross
and intergeneric cross
12
Chapter 3
Important Definitions
of Genetics
In foregoing pages of this book several technical words have been
used. It is very important todescribe those technical words Otherwise
it will not be possible to understand the subject. In addition to it
these definitions will help the students to answer several questions
asked in their practical exams.
In every cell chromosomes are present in pairs. Therefore two genes
of each character are present on these chromosomes. These genes are
expressed as capita1letter (dominant gene) and small letter (recessive
gene). For instance tallness is expressed as IT and dwarfness as t t.
At the time of reproductions pollen or semen and eggcell or ovum
are formed. In these only one gene is found which is written by only
one letter. For example tallness is expressed as T and dwarfness is
expressed as t only, in such reproductive cells.
Important technical words of genetics are explained & defined below.
Acquired Characters: Modification of character due to environment
or artificial means in life of an organism is called aquired character.
Allele or allelomorph: Each gene responsible for contrasting character
in organism is called allel or allelomorph. For example in heterozygous
plants for height, responsible gene is L and for dwarfness the
responsible gene is 1. Both these genes are called alled or allelomorph.
Ameioses: It is a special type of meiosis in which number of
chromosomes is not reduced to half as happens in normal meiosis. In
ameiosis the chromosome number remains 2 n.
Amitosis: It is special type of mitosis in which without differentiation
of chromosomes the nucleus is divided.
Autosome: Except sex chromosomes other chromosomes are called
autosomes.
Allosome: Sex chromosomes can be also called as allosome.
13
Acclimatization: Ability of organism to adjusts with changed

environment is called acclimatization.
Autopolyploid: It is that polyploid in which all chromosomes are
similar in sets.
Allopolyploid: It is that polyploid in which chromosomes are different
in sets.
Amphidiploid: It is a polyploid produced from that diploid which
has been development by cross of two dissimilar diploids.
Amphimixis: It is the mixing of male and female gametes during
fertilization.
Apomixis: Development of organism from female gamete without
fertilization with male gamete is called apomi~.
Aneuploid: Improper multiplication of chromosomes number is called
aneuploid. It is always minus or plus of a few chromosomes.
Asexual Reproduction: It is a process of reproduction not requiring
any gamete.
Asynopsis: During meiosis absence of pairing between homologues
chromosome is called asynopsis.
Autogamy: Self fertilization is also known as autogamy.
Back cross: Crossing of hybrid with any parent is called back cross.
Backcross Breeding: It is a procedure of breeding. The hybrid is
continuously (recurrent) crossed with a parent followed by selection
of the desirable character.
Balance: It is a situation when genetic factors exist in such a proportion
so that development of the organism is normal.
Basic number: It is the chromosome number of a very old member of
an organism. Basic number is expressed as x.
Biometry: Use of statistical procedures in biology is called biometry.
Biotype: Organisms of uniform genetical constitutions are called
biotype. It may by homozygous or heterozygous.
Bivalent: In first meiotic cell division, pairing of chromosomes is called
bivalent.
Breeder's seed: Seeds produced by variety developing organisation
or individual is called breeders seed. From this seed foundation seed
is produced.
14
Important Definitions of Genetics
Breeding: It is the science to change the genetic structure of plant or

animal.
Bulk Breeding: It is a procedure of breeding. Self pollinated plants
of different genotypes are grown in a bulk plot and then single plants
selection is done.
Centromere: It is a point on chromosomes on which spindle fiber is
attached. During cell division movement of chromosomes is governed
by centromere.
Character: Trait developed by gene is called character.
Chiasma: While pairing of chromosome attachment with in two
chormatid is called chiasma.
Chromatid: In beginning of meiosis each chromosomes is divided
into two threads like structures. These threads are called chromatid.
Chromomere: Chromomere are points arranged linearly in
chromosomes. Inside chromomeres gene is located.
Chromosome: Thread like structures present in nucleus are called
chromosomes. At various stages of meiosis shape of chromosomes
changes. For each kind of organism number chromosomes is same.
Clone: Organism developed by mitotic cell division or asexual
technique is called clone.
Combining ability: It is of two types-I.general combining ability. 2.
Specific combining ability.
General combining ability: In cross fertilization capacity of a parent
to combine is called general combining ability.
Specific,combining ability: Fertilization capacity of a parent is called
specific combining ability.
Covariance: Covariance is statistical unit to measure variability in a
population.
Crossing over: It is the exchange of segment between chromatids of
a chromosome. It happens because of break and join in chromatids
during meiosis.
Cytoplasm inheritance: Inheritance of characters from one generation
to other through cytoplasm is called cytoplasm inheritance. Another
name is molecular system of heredity.
Chimera: Plant part or plant made of two or more genetically different
tissues is known as chimera. Reasons may be mutation, irregular
mitosis or plastid segregation.
15
Deficiency: Removal or absence of segments or gene from

chromosome is called deficiency.
Detassel: Removal of male part from maize crop is called detassel.
Diallel cross: Crossing of several genotypes in all combination is called
diallel cross.
Dihybrid : Hybridization between male and female having two pair
of contrasting characters is called dihybrid.
Diocious: If male and female are present on different plants then it is
diocious.
Diploid: It is that organism which has two sets of chromosomes.
Diplotene: In meiosis after pachytene the stage of diplotene comes.
In diplotene each bivalent form 4 chromatids. These four chromatids
appear in pairs, but are attached at chiasma.
Disjunction: Separation of chromosomes at anaphase.
Dominance: Out of two contrasting characters, the character expressed
in Fl generation is known as dominant the phenomenon is knoWn as
dominance.
Donor Parent: In backcross breeding, the parent which gives gene to
the recurrent parent is called donor parent.
Double cross: It is the cross between two Fl hybrids.
DNA: Its full form is deoxy ribo nucleic acid. It is found mainly in
nucleus . It has adenine, guanine, cytosine and thymine.
Duplication: Double occurrence of a segment in a chromosome, is
called duplication.
Duplex: See the multiplex
Emasculation: Removal of another before dehiscence or opening of
flowers is called emasculation.
Epistasis: Dominance of gene over expression of non allelic gene is
called epsitasis. That gene whose effect is masked is called hypostatic.
Error Variance: Error caused by unknown or uncontrolled gene is
called error variance.
Expressivity: Capacity to express genetic character is called
expressivity.
Eugenics: Study of genetic for human improvement is known as
eugenics .
16
F1: Its full name is first filial generation. It is the first generation
obtains by hybridization.
F2: It is the second generation obtained on self fertilization of Fl.
F3: It is the third generation obtained on selffing of F2 hybrids.
Family: It is the group of organisms produced by one parent.
Fertilization: Union of nucleus of male and female gametes is called
fertilization.
Foundation seed: Seeds produced from breeder seed is foundation
seed. After growing it the certified seed is obtained.
Gamete: Cells formed after meiosis are called gametes.
Gene: Genes are located in chromosomes. It is made of D.N.A. and
expresses & inherits characters.
Gene Frequency: It is the proportion of expression of alternative
allele of an organism.
Gene Interaction: Gene interaction is the change in expression of a
gene by non-allelic gene.
Genetics: Study of inheritance of organism is called genetics.
Genome: Haploid set of chromosome is called genome.
Haploid: Gametic chromosome number or n chromosome number is
haplOid.
Heritability: The quantum of variability which is due to heredity is
called heritability.
Heterosis: If character of Fl is superior to parents then it is known as
heterosis.
Heterozygous: Presence of· unlike allels on corresponding loci of
homologous chromosome is known as heterozygous.
Homozygous: Presence of like allels on corresponding loci of
homologous chromosomes is called homozygous.
Hybrid: Hybrid is the organism borned by crossing parents of
different genotype:
Inbreeding: Fertilization between members of close relatives.
Inbred line: It is the near homozygous line obtained by repeated
inbreeding and selection.
Inbred-variety cross: It is the cross between inbred line and a variety.
17
Intersex: Intersex is the organism showing features of male and female.

Inversion: Rearrangement in segments of chromosome resulting
alteration in sequence of genes.
Irradiation: Exposing radiations like x-rays, gamma rays etc. to part
of organism of full organism are called irradiation. Irradiation may
cause mutation.
Isogenic lines: Isogenic lines are those lines which are uniform on all
loci.
Isoallele: Allels in which difference is not clear is known as isoallel.
Kinetochore: It is that place on chromosome where spindle fibers are
attached.
Linkage: Certain genes located very close on chromosome are passed
on together in next generation. This phenomenon is called linkage.
Such genes are known as liked genes.
Linkage value: It is the recombination fraction which indicates the
ratio of occurrence of parental type and cross over type in the progeny.
Locus: Locus is the position of genes on chromosome.
Ml, M2, M3: It indicate the number of generation after treatment
with mutagen.
Male sterility: Absence or inactivation of anther (male organ) in
organism.
Mass pedigree method: It is method of breeding where the population
collectively raised for some generations. Afterwards pedigree
selection is done.
Mass selection: It is method of breeding where desirable plants are
selected; seeds from these are collected and grown for selection.
Mating system: It is the system for sexual reproduction. It of several
types such as random i.e. any parent do mating with the other, genetic
assortative mating i.e. mating between parents of similar genetic
makeup, genetic disassortative mating i.e. mating between parents
of dissimilar genetic makeup, phenotypic assortative mating i.e.
mating between phenotypically similar parents, phenotypic
disassortative mating i.e. mating between phenotypically dissimilar
parents.
Mean: Average of several observations is known as mean.
Meiosis: It is that type of cell division where number of chromosomes
18
is reduced to half .By meiosis cell division gametes are formed.

Metaphase: It is that stage of mitosis or meiosis where chromosomes
arrive in centre of spindle.
Metaxenia: Effect of pollen on maternal tissue is called metaxenia.
Median: It is that point in observation on its both sides lower or
higher figures of observations exist.
Mitosis: It is that type of cell division which do not change number
or characters of chromosomes.
Mode: It is the value of observation in frequency distribution.
Modifying genes: These are those genes which changes the expression
of non allelic genes.
Monohybrid: It is the hybrid produced by hybridization between
parents having one contrasting character.
Monoecious: Occurrence of male and female flowers separately on
one plant.
Monoploid: It is that organism which has the basic chromosome
number.
Monosome: Deficiency of one chromosomes in diploid chromosome
compliment such as 2n-l.
Multiple Allele: More than two alternative forms of a gene is called
multiple allele.
Multivalent: Holding of more than two homologous chromosomes
by chiasmata is called multivalent.
Mutation: Sudden and heritable change in structure of gene or
chromosome is mutation.
Nulliplex: Nulliplex means that a gene is recessive on all
chromosomes. If except one or other chromosomes there is recessive
gene then it is called simplex, if recessives are on two chromosomes
then it is duplex, if recessives are on three chromosomes then it is
triple x and if recessives are on four chromosomes then it is
quadriplex.
Nullisomy: Absence of a pair of chromosome in a cell is called
nullisomy.
Pachytene~ It is the double thread stage of chromosome in meiosis.
parthenogenesis: Developm~nt of organism without fertilization of
19
sex cells is parthenogenesis.

Pedigree: Record of past generations of an organism is called pedigree.
Polysomic: If in a diploid organism there is more than two
homologous chromosomes then it is called polysomic.
Polyembryony: Occurrence of more than one embryo in seed is called
polyembryony.
Position Effect: Neighbouring gene effects the expression of the gene.
Therefore if position of a gene is changed there is a change in the
expression of the gene. It is called position effect.
Pedigree Breeding: It is a method of breeding when from a cross
between suitable parents, in the produced segregants selection is
made based on individual behaviour and pedigree record.
Phenotype: Visible form of an organism is called phenotype.
Physiological race: Physiological race is such pathogen of a species
which are similar in appearance but are different in pathogenecity.
Polycross: Independent mating in group of selected parent's
generation is polycross.
Ploygenes: These are such genes that have least effect independently
but when come together then causes variation.
Polyploidy: An organism is a polyploid which has more than two
basic set of chromosomes.
Progeny test: Study of progeny to test the genotype of the organism
is called progeny test.
Protandry: Maturity of anthers before stigma is called protandry.
Protogyny: Maturity of stigma before anther is protogyny.
Pureline: Homozygosity at all loci in chromosomes is pureline.
Qualitative Character: Qualitative character is that character which
has discontinuous variation.
Reciprocal cross: Reciprocal hybridization between male and female
parents.
Recombination: New combination of genes due to hybridization of
parents caused by crossing over and segregation is known as
recombination.
Recurrent parent: In backcross breeding the parent which is used for
repeated crossing is called recurrent parent.
20
Recurrent selection: It is a method of plant breeding. Selection is

done from segregants of desirable parents. Selected lines are further
inter crossed. Again selection of desirable ones is made. This cycle is
repeated.
Registered seed: From foundation seed the registered seed is
developed. Registered seed is further used to develop certified seed.
Recessive: From contrasting characters, the character which is unable
to express in offspring is called recessive character.
Segregation: In meiosis maternal or paternal separate out resulting
in separation of genes. It is called segregation
Selfing: Selfing is the fertilization of male and female gametes of
same plant.
Self fertility: Capacity to produce seeds on self fertilization:
Self incompatibility: On self fertilization inability to produce seed
due to some genetical reason is called self incompatibility.
Sib: Offspring of single parent is called sib. If one parent only is
common then the offsprings are known as half sibs.
Single cross: It is a cross between two genotypes.
Sport: Occurrence of mutation in vegetative bud is known as sport.
Somatoplastic sterility: Death of zygote due to disturbance in
relationship between embryo and endosperm is called somatoplastic
sterility.
Species: It is unit of taxonomy. Species is a division of genus.
Synapsis: Synapsis is conjugation of homologous chromosomes at
pachytene and zygotene stages of meiosis.
Telophase: Telophase is the last stage of mitosis and meiosis.
Afterwards resting stage comes.
Test Cross: It is the hybridization of Fl with recessive parent.
Tetraploid: A cell or organism with four sets of basic chromosomes
is called tetraploid.
Top Cross: Top cross is crossing of a selection or clone with inbred
line/variety/single cross. It is also known as inbred variety cross.
Transgressive segregation: Segregation of such offsprings which are
better or inferior than their perents is transgressive segregation.
Translocation: Transfer of segment of a chromosome on the same
21
chromosome or other chromosome is known as translocation. If it

happens mutually on two chromosomes then it is known as reciprocal
translocation.
Triploid: A cell or organism with 3 sets of basic chromosomes.
Trisomic: A cell or organism with triplication of one chromosome is
known as trisomic.
Univalent: During meiosis non pairing chromosomes are called
univalent.
Variety: It is sub division of species. A variety is different
morphologicaly or physiologicaly from other varieties of a species.
X: It means the basic chromosome number.
Xenia: It is the effect of pollen on embryo or endosperm.
Zygote: It is a cell formed by fertilization of maleand female gametes.
Zygotene: Zygotene is the stage of meiosis where thread like
chromosomes do pairing.
22
Chapter 4
Chromosome and its Structure
Body of living organism is made of cells. Nucleus is the most important
part of cell. During cell division, nucleus under goes division. At that
time the covering nuclear reticulam breaks off and in the nuclear sap
thread like structures appears. Hofmeisfer (1848) Working on
Tradescantia for the first time observed this structure in cells. Later
on Waldeyer (1888) named these as chromosome. Investigations on
chromosomes got intensified with the progress ir science of genetics.
Number of chromosome is fixed for every type of organism. During
cell division in the nuclear sap several thread like structures can be
seen. These are called chromonemata (Fig-I) (Chromonema-singular).
Every chromonemata is double.
Fig.1: Structure of a typical chromosome.
23
Every chromosome has a kinetochore or centromere which is slightly

constricted. Both sides of kinetochore are called arms. At one end of
chromosome there is another constriction- known as secondary
constriction. Upper portion of nuc1eolar organizer is called satellite
or nucleolar organizer. Some cytologists have expressed that:
chromonemata is enclosed in achromatic matrix and covered by a
membrane -pellicle. Chromosomes have a large number of
chromomeres which have genes. Now we will discuss various
components of chromosomes.
1. Chromonemata: Number of chromonemata in a chromosome may
be two or more. In some cases the number of chromonemata
goes up to 32.. Reason for increased number is the division of
chromonemata. One important thing is that whatever is the
number of chromonemata but they behave as two chronemata
only. During cell division each chromosome produces only two
chromatids.
2. Matrix: Chromonemata remain in matrix which is covered by a
covering pellicle. Both of these materials serve as protection to
chromonemata.
3. Centromere: It is one very important part of chromosome which
leads to primary constriction.
In absence of centromere, during cell division the chromosome is
unable to reach at metaphase. During anaphase normal chromosomes
move towards poles. But centromere less chromosomes lag behind.
Ultimately such chromosomes get lost resulting reduction in number
of chromosome.
Depending on the position of centromere shape of chromosome is
determined (Fig-2). If the centromere is located in centre then shape
of the chromosome is V at anaphase. If the centromere is located at
the end then shape of chromosome is I during anaphase. If the location
is sub median i.e. between end and middle then the chromosome
take J shape at anaphase. If the centromere is located at the end then
shape of the chromosome is I during anaphase. Detail study of
centromere under electron microscope was done by Depharia in 1956.
4. Secondary constriction: It is located in one or both arms of
chromosomes. Secondary constriction and satelite are related to
nucleus. Heitz (1931) reported that nucleolus is related to
secondary constriction. In polygonatum change in shape of
chromosome and little breakage resulted due to secondary
constriction.
24
(8) Telocentric (b) Acrocentric (c) Metacentric (d) Sub metacentric
Fig.2: Shape of chromosome depending on centromeric position.

5. Telomere or satellite: It is the small portion above secondary
constriction. It is also called nucleolar organizer. Some cytologists
have expressed that in formation of nucleoli at telophase stage
some material is used from chromosomes. Somehow this work
is done by nucleolar organizer. During metaphase nucleolus
disappears. Perhaps its material gets deposited on chromosome
or gets transferred to cytoplasm. Nucleolus contains dioxide
ribonucleic acid (DNA) and ribonuclic acid (RNA).
6. Chromomere: Chromomere appears like beads in chromonema.
Number, shape and location of these beads are constant in
various organisms. Balbiani (1876) and Pfitzner (1881) named
these beads as chromomere. These chromomeres have genes
which are essential unit of heredity and variation.
Special types of chromosomes:
In nature following special types of chromosomes are found.
1. Compound chromosomes:
Compound chromosomes are found in Ascaris megalocephalia . In this
organism during meiosis only one pair of chromosome is visible. But
during mitiosis the two chromosomes gets divided into numerous
very small segments. As written earlier only one pair of chromosome
is visible in meiosis. Therefore it is called compound or aggregate
chromosome.
2. Salivery gland chromosome:
Balbiani (1881) reported giant chromosomes in salivary gland. cells of
Midga larva. Afterwards Kostoff (1930), Heitz & Bauer (1933), Painter
(1933), Mechelke (1953) and Breaur & Pavan (1958) have observed
similar giant chromosomes in salivary gland cells of Drosophila (Fig 3
25
Genetics : Fundamentals and Applications
Fig. 3 a. Fruit fly Slivery gland and salivary gland chromosome under
normal mitotic
m
chromosomes at
Fig. 3 b: Salivary gland chromosomes of fruit fly (Drosophila

melanogaster)
26
a & b) and other insects. These were 100 to 200 times larger than
ordinary chromosomes. In volume these are 1000 to 2000 times more
than ordinary chromosomes. Therefore the name giant chromosome
has been given
One more speciality of this chromosome is that there are numerous
dark staining and non staining bonds & cross bonds whose sequence
is predetermined. Darkstaining bands have DNA containing
nucleoprotein but light staining on non staining bond have
nucleoprotein free from DNA.
3. Lampbrush chromosomes:
Lampbrush chromosomes are elastic in nature because by micro needle
it can be expanded several times without any breakage. On leaving,
it assumes original length. Lampbrush chromosomes were studied
by Duryee (1941 & 1950). Call (1952, 1954 & 1956) and Alfert (1954)
studied such chromosome in some fishes, birds and amphibian. Ris
(1954) has observed this chromosome in spermatocyte of some
invertebrates.
4. Accessory chromosome:
In certain organis~besides fixed number of chromosomes there are
are one or more accessory or supernumerary chromosome. These
chromosomes segregate at random. Sometimes these express non
disjunction. Occasionally accessory chromosome gets broken into
fragments. In 1905 Wilson reported accessory chromosome in cells of
Metapodius. Darlington (1935), Ostergren (1947) and Melander (1950)
have observed accessory chromosomes in several plants.
27
Chapter 5
Mitosis and Meiosis
Important Phenomenon of
Genetics
Mitosis
Growth is any living organism is because of increase in number of
cells without change in its characters. Such growth starts from zygote.
Nucleus and cytoplasm of the zygote divide and forms two cells. In
the same way each cell again divides-so the two cells forms four cells.
This process goes on till the growth of the organism is not complete.
This type of cell division is called mitosis. Due to mitosis in all the
cells number and characters of chromosomes are identical to the
mother cell. In mitosis cell division nucleus undergoes several changes.
Scientists have divided the changes in to four stages such as prophase,
metaphase, anaphase and telophase. Between mitotic cell division
there is resting stage or also known as interphase (Fig-4). All the
stages are described below:
1. Interphase or Resting stage: Chromosomes appear as interwoven
thin threads. Nucleolus and chromosomes are almost dark stained
2. Prophase: At prophase nucleus enlarges, chromosomes appear
clearer. Chromosome becomes shorter and thicker. From
beginning of prophase all chromosomes split into two. Each part
is called chromatid. The chromatids are interwoven in full length.
This interwovenship is known as relational coiling. Chromosomes
are seen scattered in the nucleus. Nuclear membrane gradually
starts disappearance.
3. Metaphase: Complete disappearance of nuclear membrane is the
beginning of metaphase. Formation of spindle takes place.
Cytologists differ on origin and number of spindle. The spindles
arrange all chromosomes in equator.
Chromatids of each chromosome are together and through the centro-
mere is attached to the spindle. The Centromere remains undivided.
28
Mitosis and Meiosis: Important Phenomenon of Genetics
If somehow it gets split even then this acts as unit only. In mitosis cell
division nucleus undergoes several changes. Scientists have divided
the ,change into four stages such as prophase, metaphase, anaphase
and telophase. Between mitotic cell division there is resting stage or
also known as interphase (Fig.4). All the stages are described below:
Chromatids of each chromosome are together and the centromeres
are attached to the spindle. The centromere remains undivided, if
somehow it gets split, even then this acts as one unit only.
4. Anaphase: At this stage centromere divide into two centromeres,
these have repulsive action. Due tothis repulsion both the chromatids
move towards poles. In absence of centromere the chromatid is unable
to move and remain at the equator.
5. Telophase: At this stage all the chromatids reach to respective
poles. New nucleus are formed at both the poles, in nucleus (plural
nuclei) nucleolus is developed. Spindle fibres disappear.
Chromosomes loose the capacity to get stained.
(a) Interphase
•
~ (1) Mitotic telophase (b) Early mlotIc
o prophesQ I\1
(d) MitotIC metaphaSe
Fig.4: Diagramatic representation of various stage of mitosis.

29
6. Cytokinesis (Division of cytoplasm): Cytokinesis starts fonn

cell plate fonnation. In the end of telophase in the middle of
cytoplasm spindle fibers make fragmoplasts. Several fragmoplasts
join and fonn a plate. Length of this plate increases and joins
with cell wall. The plate is called cell plate. Over this cell plate
pectin deposition starts. Afterwards this cell plate is known as
middle lamella. From both side of middle lamella secretion of
some compounds (ploysacharide) takes place. Afterwards this
wall is called primary wall. Again deposition of cellulose,
hemicellulose and polysacharide take place. Afterwards the wall
is called secondary wall. This is the end of cylokinesis and the
cell gets divided into two cells.
In animals cytoikenesis take place. in different way. It takes place by
furrowing of the cytoplasm.
Significance of mitosis:
An important significance of mitosis is that the chromosomes of all
the produced cell are identical to the mother cell. Chromosome
number and their characters are just the same of the mother ce,ll. Due
to increase in mitotic cell division the organism achieves growth.
Mitosis is very important phenomina for vegetative propagation.
Meiosis:
Meiosis cell division is also known as reduction division and
heterotypic cell division. Gamete formations are due to meiosis only.
Meiotic cell division results in reduction of chromosome number to
half.
Meiosis occurs in two phases ie. two times prophase and two times
metaphase, two times anaphase and two times telophase. Therefore
stages of first meiosis are called prophase-1 metaphase-1, anaphase-1
and telophase-1.With telophase-1 first meiotic cell division completes.
The second meiotic cell division starts. The stages are prophase-II,
metaphase-II, anaphase-II and telophase -11. With telophase-II the
nucleus divided two times and the meiosis (Fig-S) finishes. Meiosis is
discussed in detail below.
First meiosis:
1. Prophase-1: During prophase-1 various changes in chromosomes
can be described in five stages as given below.
30
a. Leptotene or Leptonema: Cell size increases, chromosomes

are thin and long. Presence of chromomere is clear.
b. Zygotene or Zygonema: At this stage pairing or synopsis in
chromosomes takes place. Every living organism has one
haploid chromosome set from father (sperm or pollen)
and one haploid chromosome set from mother (egg).
Chromosomes obtained from father matches from
chromosomes obtained from mother except to sex
chromosome. Therefore in every diploid cell there are pairs
of chromosomes (resembling to each other) obtained from
father and mother, are called homologous chromosomes.
At zygotene stage of meiosis these homologous
chromosomes come together and form pairs. This process
is called synopsis or pairing. In this process centromeres of
homologous chromosomes come opposite to each other.
Pairs of homologous chromosomes are called bivalents.
Therefore if number of chromosomes is 2 at leptotene then
at zygotene it appears as one bivalent.
c. Pachytene or Pachynema: At pachytene every chromosome
of bivalent, splits longitudinally into two chromatids. But
the chromatids are united at centromere. Chromatids do
not split. Therefore each bivalent appears to have four
strands at pachytene. Nucleolus is visible at'this stage.
d. Diplotene or Diplonema: At diplotene stage homologous
chromosome start movement from each other. But the
homologous chromosomes are not completely separated i.e.
those are connected at one or more places. If connected at
one place then the bivalent appears to be a cross. If connected
at two places then bivalent has one loop. If connected at
more places then the bivalent has more loops. Each place of
connection in a bivalent is called chiasma (plural chiasmata).
Number of chiasma and its location depends on length of
chromosome and the type of species. Even the smallest
, bivalent has one chiasma. Depending on location chiasm is
of two types.
(i) Terminal chiasma: It is located at the end of chromosome.
(ii) Interstitial chiasma: Instead at end, the chiasma is located
any where else on the chromosome. With the separation of
homologous chromosomes interstitial chiasmata can get
converted into terminal chiasma. Theprocess is Simple. Due
31
to movement in centromeres the homologous chromosomes

moves apart. So interstitial chiasma moves towards end of
the chromosome. Thus ultimately it becomes terminal
chiasma. This movement of chiasma is known as
terminalization. At diplotene stage length of chromosomes
gets reduced but their thickness increases.
e. Diakinesis: At diakinesis chromosome appears short and
thick, Nucleolus starts disappearance and completely
disappears by the and of diakinesis. Most of bivalents
specialy smaller ones because of terminalization assumes
nail or similar shapes,
2. Metaphase:
Like metaphase stage of mitosis, in meiosis also at metaphase -1 the
nuclear membrane disappears.Spindle fibres appears. Some cytologists
give the name prometaphase to the phases between disappearance of
nuclear membrane and formation of spindle. All bivalents come on
the equator.
There is a difference between metaphase of mitosis and metaphase-1
of meiosis During metaphase of mitosis every chromosome has a
divided centromere and all the chromosomes lie on equator. Where
as in metaphase-1 of meiosis each bivalent has two undivided
centromers and bivalents do not just lie on equator but their
centromeres appears to be diverted towards poles.
3. Anaphase-l:
At ananphase-1 of meiosis bivalents do not have divided centromere,
due to which homologous chromosomes along with two chromatids
moves towards their poles in contrast to mitotic anaphase where
centromeres are divided and chromatids goes towards poles The
result is that at end of anaphase-1 on the poles there is only haploid
number of chromosome, In other words only half the number of
chromosomes at the poles.
4. Telophase-l:
Telophase -1 is the last stage of first phase of meiosis. It is for a short
time only. Nuclear membrane appears. Spindle fibers disappear. These
nuclei are called dyad. Dyad contains only half the number of
chromosome in the organism. In certain organism such as Trillium
the chromosomes after reaching the poles withoutcentering telophase-
1 attains second phase of meiosis.
32
Second Meiosis:
All stage of second meiosis is similar to mitosis except the differences
mentioned below.
a. In comparisom to mitosis, second meiosis has only half the
number of chromosomes.
b. Chromatids remain at some distance from each other. Hence
there is no relational coiling.
c. In mitosis there is no change in the genetic sequence, where as
during first meiosis due to chiasma (plural chiasmata) there is a
change in genetic sequence at second meiosis.The degree of
changes depend on number of chiasma formed.
Stages of second meiosis are mentioned below:
a. Prophase-II: It is a small stage. Nuclear membrane appears and
chromosomes appear as a net work.
b. Metaphase-II: At this stage nuclear membrane disappears.
Spindle fibres appear. Chromosomes come on the equator and
their centromers get connected with the spindle fibres.
c. Anaphase-II: Centromers get divided. Both the chromatids of
chromosomes start movement towards respective poles.
d. Telophase-II: At this stage chromatids reach to the respective
poles and form net like structure there. Nuclear membrane
gradually appears. Cell plate forms and due to this four cells are
formed. Each cell has a nucleus. Number of chromosomes is only
half (n) in all the four nuclei.
Cytokinesis in meiotic cells:
Cytokinesis (division of cytoplasm) mechanism varies in different
organism. In some organism during first meiotic division a wall is
formed in middle of cell division, dividing the cell into two cells. In
second meiotic division another wall get made at right angle to the
first wall. In this way the meiotic cell gives rise to four cells. In some
other organism during first meiotic division cytokineSis do not take
place. In the end of second meiotic division two walls get made in
the cell dividing it into four cells. Each cell have only half the number
of chromosomes.
33
Significance of meiosis:
1. Meiosis helps in continuation of chromosome number in
organismS: Due to meiosis the chromosome number gets reduced
to half in gametes. when male and female gametes (with half
number of chromosomes) unite and fertilize then the zygote gets
the complete number of chromosomes. In this way the
chromosome number remain constant
2. Helps to know breeding behaviour of organism. Investigations
of meiosis helps to understand breeding behaviour of the
organism.
Fig. 5: Diagrammatic representation of various stages of meiosis.
34
Differences in mitosis and meiosis are given below:

Diffrences in mitosis and meiosis
Mitosis Meiosis
1.Nucleus divides once 1. Nucleus divides twice.
2. Prophase is short duration. 2. Prophase has long duration there are
There is no substage substages such as leptotene, zygotene,
pachytene, diplotene and diakinesis,
3. There is no pairing in 3. Homologous chromosomes pair and
homologous chromosomes form bivalents
4. Chiasma formation is absent 2. Chiasma formation occurs therefore
chromatids exchange some parts.
5. At anaphase the 5. At anaphase -1 centromere do not divide.
chromosomes along with In each bivalent the two chromosomes
centromere divide &: move move towards the opposite poles along
towards opposite poles and with their chromatids at the poles.
maintain 2 number of Chromosome number is reduced to
chromosomes. half.
6. At telophase both the 6. At telophase-1 both the daughter cells
daughter cells have 2 n have only n chromosome number.
chromosome number
7. There is no second division 7. There is second division of nucleus. In
of nucleus. the second division centromere also
gets divided. Thus second division is
matching to mitosis. But instead of 2n
there is only n number of chromosomes.
8. In end of mitosis neither the 8. In end of meiosis number and character
number of chromosomes of chromosomes are changed.
nor the characters of
chromosomes change.
9. Two cells are formed. 9. Four cells are formed .
10. Mitosis occurs in only 10. Meiosis occurs in only reproductive
somatic cell. cells.
Spermatogenesis and Oogenesis
Spermatogenesis result in sperm formation and oogenesis result in
ova formation in animals. Therefere this knowledge is also essential
to students of genetics. There is similarity in these two processes.
But in some matter they differ.
These processes occur in animals when they achieve sexual maturity.
In certain mammals such as human being once this process starts it
continue till he or she becomes sexually inert. In other animals these
35
processes occurs only in their breeding season.

Testes of animals have numerous tubules which are united with
germinal epithelium. Every cell of this epithelium acts as
spermatogonium. Similarly every cell in epithelium of ovary acts as
Oogonium. Spermatogonium and oogonium grow by mitosis, each
have somatic chromosome number. On reaching sexual maturity all
spermagoniums produce primary spermatocytes. But in ovary at one
time only one oogonium results primary oocyte which is much bigger
as compared to primary spermatocyte. First meiosis occurs in primary
spermatocyte and primary oocyte. Due to this action primary
spermatocyte gives rise to two secondary spermatocyte.
Incase of primary oocyte there is uneven division resulting one big
secondary oocyte and one small polocyte or polar body.
Second meiosis occurs in secondary sepermatocyte and secondary
oocyte .. Secondary spermatocytes result four spermatids. Secondary
oocyte gives rise to ootid and polocyte.Each spermatid undergoes
changes then it becomes sperm.
(a) Nucleus combines with some cytoplasm and forms head.
(b) Golgi bodies accumulate in anterior tip and form acrosome which
help in penetration of ovum.
(c) Mitochondria accumulate in middle piece which gives strength
to sperm
(d) Remaining cytoplasm takes the shape of flagellum or tail which
helps in movement of sperm.
In contrast to spermatids ootid straight way develops in ovum. Almost
full cytoplasm of primary oocyte comes in ovum. Polycyte disappear.
Number of chromosomes is only half (n) in sperm and ·ovum.
Differences in spermatogenesis and oogenesis are mentioned below:
Differences in spermatogenesis and oogenesis:
Spermatogenesis Oogenesis
1. Several spermatogonia divide and 1. At one time only one oogonium
forms sperm divides to from ovum (egg.)
2. One spermatogo-nium produces 2. One oogonium produces only one
four sperms. ovum (egg).
3. Sperm is minute. 3. Ovum (egg) is quite bigger than
sperm.
36
Chapter 6
Laws of Inheritance
To understand genetics properly, we have to understand the laws of
inheritance.These laws are based on breeding experiment on pea done
by the famous scientist John Gregor Mendel (native of Austria) who
lived from 1822 to 1884. By this time it was clearly understood that
transfer of characters from one generation to other generation can
occur only through reproductive cells (gametes). One more thing was
clearly understood that characters achieved by environment or
artificial means can not be transferred from one generation to other.
Due to these facts John Gregor Mendel and other scientists have
expressed that phenotypic appearance of any organism depends on
its genetic constitution (genotype). One thing more, change in
phenotype does not have any effect on its genetic structure
Based on above mentioned facts John Gregor Mendel and other
scientists did several experiments on plants and animals. Before John
Gregor Mendel several scientists have done experiments but were
unsuccessful because of adopting faulty methodology. Therefore
seeing all this John Gregor Mendel very carefully adopted the right
method and the material. Those precautions are detailed below.
1. The organism selected for the experiment should be homozygous.
If it is not homozygous then after several generations of selfing
it should be made homozygous.
2. For crossing only those parents should be selected who possess
contrasting characters.
3. At one time only one character should be investigated.
4. For the experiment the organism should be available in plenty
and it should have short life-cycle.
5. Record of observations should be clear and complete.
John Gregor Mendel discovered the principles of genetics in 1867 but
it came to light after a long period. Later on De Veries in Holland,
Correns in Germany and Tschermark in Austria have worked on this
aspect, then in 1900 they have brought out the discovery of John
Gregor mendel .
37
Genetics: Fundamentals and Applicatiofls
Material and method of John Gregor Mendel:

Keeping in mind all the above mentioned precautions John Gergor
Mendel preferred pea (Pisum sativum ) for his research work. Pea
has all the desired attributes because:
1. Varieties of peas have contrasting characters. Following 7
characters were studied by John Gregor Mendel.
Character Details of character
1. Seed shape.......................... Smooth & Wrinkle
2. Cotyledon colour .................... .. Yellow & Green
3. Seed coat colour .................... .. Coloured & White
4. Pod type ............................ .. Hard & Soft
5. Pod colour ............................ . Yellow & Green
6. Flower position ...................... .. Axillary & Terminal
7. Stature ........... : ..................... .. Tall & dwarf
On intensive studies of these characters Gregor Mendel has observed
that characters written first in column 3 are dominant over characters
written later in same column. Various experiments of John Gregor
Mendel have revealed some important phenomenon which became
well known as principles of heredity. These principles are followings.
1. Law of dominance.
2. Law of segregation.
3. Law of independent assortment. These laws are discussed below:
Law of dominance:
On performance of breeding experiments on peas and other organisms
John Gregor Mendel has observed that in F1 offspring characters of
only one parent appear. On Studies of contrasting characters these
observations were confirmed. Therefore the law of dominance was
put forth. When homozygous chromosomes having contrasting
characters are crossed then in the F1 hybrid the appearing character
is called dominant and the character not appearing in F1 hybrid is
known as recessive. Based on results of breeding experiments on
seven contrasting characters of pea Gregor Mendel has listed dominant
and recessive characters as given below.
38
Laws of Inheritance
Dominant Recessive
1. Smoothness of seed Wrinkleness of seed
2. Coloured seed coat White seed coat
3. Yellow cotyledon Green cotyledon
4. Hard pods Softpods
5. Green pods Yellow pods
6. Axillary position of flowers Terminal position of flowers
.
7. Tall plants Short plants.
After proving the law of dominance in monohybrid crosses Gregor

Mendel did several dihybrid crosses and again found the truthness
of law of dominance. This can be explained by following example.
Examples to explain Law of Dominance:
1. Monohybrid crosses (Cross between one pair of contrasting
character): Homozygous purple flowered pea plants were crossed
with homozygous white flowered pea plants (pollen from one
plants was transferred on stigma of other plant). The seeds
obtained were sown & cultivated. These plants yielded pink
flowers. So the pink character was expressed as dominant and
white character was expressed as recessive (Fig-6).
2. Another example to prove law of dominance is a monohybrid
cross between white cow and red cow. The F1 generation has
only roan colour showing dominant red over white colour rat.
(Fig- 7)
Cytological explanation of the law of dominance:
It has been clear that in every cell (Somatic or Gonad) there are
homologous chromosomes in the nucleus, every chromosome has
numerous genes. At reproductive phase one of the chromosomes from
pairs gets transfered to gametes. The process is explained below.
In black rat the genes responsible for black colour in the homologous
chromosomes can be designated as BB In Brown rat in the homologues
chromosome the genes responsible for brown colour can be
designated as bb At reproductive phase from BB by meiosis one of
the B comes to sperm. In the same way from bb by meiosis one of the
b comes to ovum (egg) of the rat, at the time of crossing the sperm
with B unite & fertilize with ovum having b and forms the zygote Bb.
39
Fig.6: Monohybrid cross between Fig. 7: Monohybrid cross between

pea flowers showing dominance white cow and red cow showing
of character in Ft. Law of dominance in Ft.
Because black is dominant therefore B express the black colour though

b coexist.
Incomplete dominance:
For testing the law of dominance several scientists have done many
experiments on several plants and animals. They observed that law
of dominance is not always true. In some case dominance is
incomplete. The result is that characters of F1 generation do not match
with any parent (Fig-8). Examples of such phenomenon are found in
4 o'clock (Mirabilis jaZapa), snap dragon and australian fawl.
In case of 4 O'clock plant (MirabiZis jaZapa) when red flower type was
crossed with white flower type then in the F1 hybrids neither red
nor white flowers were present. Instead all the plants expressed light
red flowers only. When light red flowered plants were selfed and F2
generation was raised then the F2 generation have revealed three
colours - red, light red and white in the ratio bf 1: 2: 1. Where as, as
per the law of dominance only red and white flowers should have
40
Laws of Inheritance
Fig.8: Incomplete dominance in 4 0'dock plant <Mirabilis jalapa)

appeared in ratio of 3:1. Therefore the above mentioned phenomenon
was called incomplete dominance.
Law of Segregation:
John Gregor Mendel has selfed F1 generation and rose F2 generation.
He observed that most of the F2 generation expressed the character
of one dominant parent or the character expressed by F1 generation
and the character of one recessive parent. In the F2 generation these
characters always expressed in ratio of 3:1 i.e. 3 dominant and 1
recessive. Based on these observations Gregor Mendel has put forth
the law of segregation. It is explained below:
When two alleles of contrasting characters (one is dominant and the
other recessive) are brought together by hybridization in F1
generation then those contrasting alleles do not blend, interact or
affect each other. When F1 forms gametes then these alleles
(characters) separate freely and go freely in gametes. Afterwards
41
they reach freely in next generation. This law has been expressed as
law of segregation. In one gamete allele for either pink colour or for
white colour can go. Both these characters can not go in same gamete.
Explanation for law of segregation:
1. Law of segregation can be understood well by hybridization in
pea (Pisum sativum ). Homologous pink flowered variety was
crossed with homologous white flowered variety. In the F1 all
the plants expressed pink colour, showing it is a dominant
character. When the F1 was selfed and raised to F2 generation
then F2 generation flowered in the ratio of 3 purple : 1 white
according to law of segregation (Fig-9)
,~
J._=- ~-=-
DO
,...
N Dd It!
,...,... o-t
DD
,... T"
OIl
,...
N It!
DIwt
Fig.9: Hybridization between pink and white flower pea showing,the

law of segregation in Ft and F2 generation.
42
Laws of Inheritance
Law of independent assortment:

On achieving success in experiments on monohybrid crosses John
Gregor Mendel experimented on dihybrid crosses i.e. on two
contrasting characters. In case of pea (Pisum sativum) out of seven
contrasting pairs he chosen two contrasting homozygous characters,
for example round & yellow and wrinkled & yellow seeds. On
hybridization of these two contrasting characters as expected Gregor
Mendel in F1 found all hybrid plants bearing round and yellow seeds
only because these two are dominant characters. When this Fl
generation was self crossed and F2 was produced then the phenotypic
ratio was not 3:1 but it was 9:3:3:1 - seeds were of four types. Gregor
Mendel observed that out of four types two types were totally new
types. Based on these facts he has formulated a new law - the law of
independent assortment.
Mendel explained that various characters are totally free from each
other. The character form of seed is totally free from colour of seed.
Any form can be with any colour. Any colour can be with any form.
Segregation at F2 therefore is fully free.
The law of independent assortment can be explained cytologicaly.
Two or more than two contrasting contractors are totally free from
each other because in every cell there are separate pairs of
chromosomes. In every chromosome there are several genes (each
gene is responsible for one character) each pair of chromosome is
totally free. Therefore at the time of gamete formation during meiosis
every pair of chromosome is totally free and during metaphase-1 get
attached to spindle as per their wish. If suppose there are two pairs
of heterozygous chromosomes then at meiosis those can get arranged
in four types on the spindle. If there are three pairs of heterozygous
chromosomes then at meiosis those can get arranged in eight types
on the spindle (Fig-lO)
OtAef example with cytological explanation of law of independent
assortment
Law of independent assortment can be explained by experiment on
pea having two contrasting characters. Suppose the characters are
.smoothness & yellow seeds and wrinklness & green seeds.
Smoothness & yellow is dominant over wrinklness & green. For
smoothness the responsible genes are SS. For yellow seeds the
responsible genes are YY. Genes responsible for wrink1eless are ss
and genes responsible for green seeds are yy (Fig-ll).
43
Fig.IO: Process of assortment of chromatids during gamete formation.
•
-,-
.ssyl' •
--
_w
P,;
- I
-
G>
I
•
®
Ft; -,...,.
$,,">'
. .-
,,.® ,14@ 1/4€) lf4®
ssrre
Ill.::: .ssl'".
-.
F"
1f4@
SoW • SoY".
- 1f4@
SfY1. .ssw.
1116= 1/16=
Sol'" •
1/16=
1/16= Ill'=-
4t .,.,.-
1/1'=
Sowe SI,.,..1/" :::'"
=..
.
.YIf
tf4@)
1116= Ill'=- ..
" :",
SIw.
1/..
So,.,. • ,.,. -
tl4®
.".= w;:'" 1/"=:'
Fig. 11: Dihybrid cross (seed shape and colour) of pea seeds showing
1/" :::""
impendent assortment.
44
Laws of Inheritance
If pea plant with SSYY genome is crossed with ssyy genome, then in
F1 generation all the plants will have SsYy genome Therefore their
seeds will be round and yellow. The reason was that by meiosis in
the plant with SSYY genome SY reached in one gamete and SY reached
to other gamete. Similarly in the plant having ssyy genotype by meiosis
sy went to one gamete and sy went to other gamete. "For fertilization
any SY gamete can unite with any sy gamete. Therefore SsYy genomed
heterozygous F1 is produced whose seeds are round and yellow. In
the F1 during reproduction stage in male and female gametes four
type of genetic combination may take place. First possibility is that 5
combines with Y and form SY. Second possibility is that S combines
with y and form Sy . Third possibility is s combines with Y and form
sY. Fourth possibility is that s combines with y and form the gamete
with sy genetic combination. Similar genetic combination will happen
in female gametes.
Therefore if the hybrid plant with SsYy genome is self fertilized then
pollen of four kinds are produced -Sy,sy,sY and sy. Similarly ovules
of four types are produced i.e.-SY Sy, sY and sy. These pollen and
ovules can combine in sixteen ways. These can be mentioned below:
1. SSYY (smooth & yellow)
2. SsYy, SsYY,SsYy,SSYy,RrYy,SsYY, SsYy,SsYy (all smooth & yellow)
9
3. SSyy (smooth & green)
4. Ssyy, Ssyy (all smooth & green) 3
5. ssyy (wrinkled & green) 1
6. ssYY (wrinkled & yellow)
7. ssYy, ssYy (wrinkled & yellow) 3
In this way 9 smooth & yellow seeded, 3 smooth & green seeded, 3
wrinkled & yellow seeded and 1 wrinkled & green seeded pea
plant were obtained. It is the law of independent assortment.
Study of dihybrid cross by checkerboard
Above explained dihybrid cross can be also understood very well
by checker board (Fig-12.) On the horizontal side various types of
male gametes are written. On vertical side various types of female
gamete are written. Afterwards every male gamete is fertilized with
every female gamete. The results of fertilization are written in the
corresponding squares. All the squares are filled up, and then the
phenotypes are separated based on following principles:
45
Genetics : FuntlRmentals and Applications
1. Dominant gene is written in capitalletter. Wherever itis presented

(double or single) it expresses phenotypically.
2. Recessive gene is expressed in small letter. This gene expresses
phenotypically when present in double. For example in the
genome Rryy letter y mean green seed, But the seed is green
only if there are two y genes. In other words in case of Yy the
letter y is unable to expresses itself because of the presence of
only one y gene.
RY Ry Ry Ry
RY RRYV RRYy RrYV RryY
Ry RrYy Rryy RrYy Rryy
rY RrYV RrYy RrYV RrYy
ry RrYy Rryy rrYy Irryy
RRYY (round & yellow)

RRYy, RrYy, RrYy, RRyY, RryY, RryY. RrYy, RryY, (round & yellow)9
Rryy, Rryy, Rryy (round & green) 3
RrYy, rryY, rryY (wrinkled & Yellow) 3
rrYY ( wrinkled & green) 1
Phenotype ratio 9:3:3:1
,......
........
• •
~=,'::t=-::"M "
v..................... '
"...... \
RNy
~RNy
114,1fY.
~~
,...,. ......
t,~ ~/4,!!. __~~_'!-'_'i
y ................._ . . . . . . -
!14/fY, .. I • ! .1 -. !
/ : ilffr : ~'i W'IY t M)' :
--tr. '~1~1¥~
(
i• i
~
114,y, • !• t •
,
:
fWY ! Rr'tY---;--
,~~,~ '-~t- ~----
' IJ'VY 1 IfI)o
------r- "'. :
,..."~ '~;
1I4ty: • i• :• 1 • !
• ,Rr'tY i Rtyy i ,""1 j "»' :
...... ~ tlfI8"'Y- : tnGR-w : tnGtrY- : 1118"".
~_~ W1eo : tn8. : SNS. : 111"
Fig. 12: Dihybrid cross (seed shape and colour) of pea seeds showing
impendent assortment.
46
Laws of Inheritance
Exceptions to the Law of independent assortment:

After John Gregor Mendel several scientists have observed that in
some organisms two or more genes pairs do not show the law of
independent assortment. Among such scientist some important ones
are Bateson, Punnet and Shull. These and others have recorded
following observations.
Exception 1 (Duplicate factor):
In shephered purse plants when triangular seeded and round seeded
varieties were crossed in F1 generation all plants revealed triangular
seeds. This F1 was self-crossed to produce F2 generation. In F2
generation triangular and round seeded plants were recorded in the
ratio of 15:1. Scientists have explained that this ratio was because of
duplicate factor ( gene) . Duplicate factor can be defined as these are
such factors (genes) which express parallel affect. It can be understood
by following cross.
Triangular seed x Round seed
TT RR ttrr
Tt Rr-----F1
Triangular seeds.
Selfed
9TR 3Tr 3tR 1tr -F2
Triangular Triangular Triangular Round
Seed Seed Seed Seed
It can be also written as:
Triangular seed=9+3+3=15
Round seed= 1
Hence phenotypic ratio =15:1
Genotypic ratio =9:3:3:1
Exception 2 (Inhibitory factor):
In poultry when coloured feathers were crossed with white feathered
poultry, in the F1 generation all poultry revealed white feathers. After
selfing the F1 generation the F2 generation was produced. F2 revealed
13 white feathered poultry and 3 coloured feather poultry. So the
ratio was 13: 3 . This ratio was the result of inhibitory factor (gene).
47
Inhibitory factor ( gene) is that gene which does not express its effects
but inhibits the affect of other gene. This phenomenon has been
expressed by following experiment
White feathered fowl x coloured feather fowl.
WWcc wwCC
WwCc
White feathered fowls
Selfed
9WC 3Wc 3wC lwc
white feather white feathers coloured feathers white fethers
It can be also written as:
white feathers: 9+3+1=13
coloured feathers: 3
Hence, phenotypic ratio=13:3
Genotypic ratio=9:3:3:1
In white feathered fowls there is W gene but this gene independently
does not produce any white colour in feathers. Moreover this W gene
(factor) inhibits the expression of coloured feathers by the gene C.
Exception 3 (complimentary factor)
When two white flowered varieties of pea were crossed then in the
Fl generation all the plants had coloured flowers. On selfing the F1
generation F2 was produced. In the F2 9 coloured flower plants and
7 white flower plants were obtained. This change was the result of a
new phenomenon - complimentary factor (gene). It is that gene which
singly express one character but when exits together then a new
character is developed. It will be easily understood by following cross.
White flowered pea x White flowered pea .
AAbb aaBB
~~-----------------n
Coloured flowered pea
Selfing
9AB 3Ab 3aB lab
Red flowered White flowered white flowered white Flowered
48
Laws of Inheritance
Red flowered pea =9

White flowered pea 3+3+1 =7
Therefore phenotipic ratio = 9:7
Genotipic ratio = 9:3:3:1
In this cross when A and B are present separately then on
hybridization only white flowered pea is produced. But when A and
B come together then it produces red flowered pea.
Exception 4 (supplementary factor) :
In Sorghum vulgare there are two main types e.g.(1) blackish- purple
glumes and (2) brown glumes. When blackish - purple glumes type is
hybridized with brown glume type then in F1 generation plants the
colour of glumes was quite different - it was reddish purple. On Self-
fertilization of F1 plants F2 was produced. In F2 plant 9 had reddish
purple glumes, 3 had blackish - purple glumes , and 4 had brown
purple glumes. Hence the phenotypic ratio was 9:3:4 This ratio
happened because of supplementry factor (gene).!t is those genes
(factors) which separately express different characters but when they
are together then they express new character . This can also be
understood by following cross.
Blakish purple gulme x Brown glume
Sorglum vulgare Sorghum vulgare
AAbb aaBB
AaBb
Reddish brown
Self fertilization
9AB 3Ab 3aB lab Reddish

purple Blakish purple Brown Brown
It can be mentioned like following:
Reddish purple = 9
Blakish purple = 3
Brown = 4
Hence Phenotypic ratio = 9:3:4
Genotypic ratio = 9:3:3:1
49
As mentioned in the cross in case of brown glumes there is a gene B

which does not has any expression but has some connection with
brown colour. But when this B unite with other gene A (which express
blackish purple color) then a new color reddish purple is produced.
Hence B is supplement to A. Therefore on uniting B with A only
reddish purple color develop.
Exception 5 (Polymorphism factor or additive gene action):
In some varieties of wheat (Triticum aesativum) colour of seed is deep
red and in some varieties it is whitish. When deep red wheat is
hybridized with whitish wheat then the Fl generation expressed deep
red coloured wheat. Fl was self fertilized to produce F2 generation.
F2 revealed 9 deep red, 6 light red and 1 white seeded plants. So the
phenotypic ratio was 9:6:1. This change happened due to
polymorphism factor ( gene), inotherwords it is additive gene action.
It can be understood by following cross:
Deep red seeded wheat x Whitish seeded wheat
AABB aaBB
AaBb
Deep red seeded wheat
Self fertilization
9AB 3Ab 3aB lab

Deep red Light red Light red Whitish
It can also be written as:
Deep red = 9
Light red = 6
Whitish = 1
Hence, Phenotypic ratio = 9:6:1

Genotypic ratio - 9:3:3:1
In this example genes A and B separately produce light colored seed.
But when come together they intensify the wheat color of deep red,
such genes are called polymorphism gene (factor) or additive gene
(factor)
50
Laws of Inheritance
Exception 6 (Epistasis)
In Sorghum vulgare (Jowar) when red nonpearly grained variety was
hybridized with white pearly variety then in the Fl generation all the
plants expressed red non pearly grain. Fl were self fertilized to
produce F2. It was found that twelve plants expressed red non pearly
grains, three plants expressed white pearly seeds and one plant
expressed white chalky (without pearly character). This behaviour
happened because of epistasis gene (factor). Masking effect is called
epistasis. So the gene (factor) which masks the effect of other gene is
called epistasic gene (factor), and that gene whose expression is
masked is called hypostatic factor (gene). This phenomenon can be
understood by the following cross.
Red nonpearly seeded jowar x White perly seeded jowar
AAbb aaBB
AaBb
Red nonpearly seeded jowar
Self fertilised
9AB 3Ab 3aB lab

Red nonpearly Red nonpearly White pearly White chalky
It can also be written as:
Red non pearly = 12
White pearly = 3
White chalky = 1
Hence, Phenotypic ratio = 12:3:1
Genotypic ratio = 9:3:3:1
In this cross in the red grain variety there is a gene for pearliness but
it has been masked by A gene (epistatic gene or factor).
Causes of these exceptions.
Experiments of later on have suggested that the causes of these
exception are mutation, crossing over and gene interactions. These
phenomenons are explained lateron.
Method to know genotype of an organism
Whether a character is homozygous or heterozygous, to know it John
51
Gregor Mendel has explained a method - known as test cross. Its

details are given below.
TestCross:
The organism to be examined is crossed with homozygous recessive
parent. By study of F1 the genotype of the organism is found out as
mentioned below:
1. If in F1 all the organisms are showing recessive character then
the organism is homozygous recessive.
2. If in F1 all the organisms are showing dominant character then
the organism is homozygous dominant.
3. If in F1 some organisms exhibit dominant character and some
organisms exhibit recessive character then the organism has
heterozygous genotype.
Explanation for test cross:
The above mentioned facts are based on a truth that heterozygous
monohybrids produce two types of gametes. Whereas homozygous
monohybrid produced only one kind of gamete. Therefore on crossing
W l~
SS • •
• • , ........"
.........
I
I
•
I
SI
tMOth
...... I i .......I
..... a
I
.......
I
•(9
!~ Ss •
e] !,
1*......
•
••
1 ...... :
1 ........
Fig. 13: Test cross of a sample of smooth & wrinkled pea seeds
52
Laws of Inheritance
of heterozygous recessive with homozygous recessive parent (test

cross) two types of offsprings are produced. Whereas homozygous
dominant when crossed with homozygous recessive only one type of
offspring is produced.
To explain above the example of summer squash plant be taken. In
two varieties white & disc shaped fruits and yellow & spherical fruits
are produced. We have to find out the genotype of 3 samples of white
& disc fruits. Plants produced from these seeds were crossed with
yellow & spherical fruited plants (because it is homozygous recessive).
(A) If one sample is crossed with yellow & spherical and the Fl
generation reveals white & disc fruits, then the sample is homozygous
dominant ( Fig-13).
Assume smooth seed are controlled by the gene (factor) 5 and wrinkled
shape is controlled by the gene s.
Backcross
Back cross is that breeding experiment where Fl generation is crossed
with homozygous parent. If the Fl is crossed with homozygous
recessive then it is called test cross and also back cross. But if the Fl
is crossed with only dominant homozygous parent then it is called
only back cross.
On crossing Fl with homozygous dominant parent all the resultant
plants resemble to dominant parent but their genotypic ratio is 3
heterozygous dominant and I homozygous dominant
Bull of either breed

Backcross progeny
e.g. Breed A (100%)
(75%A - 25%8)
fir}
Bull of Br89d A (1CIO%) Market all animals.
F1 AB female progeny (50 .... 50%)
Market all Ft AB male progeny. Ft AB fiIImaIN

cen be brad on !he ~ or btqhl In
turn prqMlrlias uaiIg "'0 CIOII bl1led&
Fig.14: Back cross.

53
If Fl is back crossed to homozygous recessive parent then half of the

resultant population has dominant character and other half population
has recessive characters (Fig-l4).
Trihybrid Cross:
If a parent having three dominant homozygous parents is hybridized
with the parent having three recessive homozygous then such a
hybridisation (cross) is called trihybrid cross and such a hybrid is
called trihybrid. This difficult cross can be well understood by Gregor
Mendels law of independent assortment.
Yellow, round seeded & red flowered plants of pea was crossed with
green, wrinkled seeded & red flowered pea. Genotypes of these
parents are RRYVCC and rryycc . From these parents the gametes
will be RYC and ryc. On hybridization the Fl hybrid will have the
genotype RrYyCc and the phenotype round, yellow seeds with red
flowers. This Fl will form eight types of male and female gametes
such as RYC, RYc, RyC, Ryc, rYC, rYc, rye & ryc . Such male and
female gametes will fertilize in 64 combinations. Phenotypic ratio of
F2 will be 27 round yellow red ; 9 round yellow white; 9 round
green red ; 9 wrinkled yellow red ; 3 wrinkled green red; 3 wrinkled
yellow white i 3 round green white & 1 wrinkled, green white. So
there are 8 types of phenotypes. Genotypes are of 27 types.
Polyhybrid cross:
A cross between parents having more than three contrasting characters
is known as polyhybrid cross. In polyhybrid cross the number of
genotype and phenotype become quite high. Following table gives
number of contrasting characters (gene~), number of genotypes
formed in Fl, number of genotypes formed in Fl, number of
phenotypes in F2 and possible number of combinations in F2 are
given.
Table: Relation between number of contrasting characters, types of
gametes formed for Ft, number of phenotypes in Ft and
number of possible combination in F2
No.of contrasting Type of gametes Type of phenotypes Possible combination
characters forFl inFl inF2
1 2 2 4
2 4 4 16
3 8 8 64
4 16 16 256
·5 32 32 1024
n 2n 2n 4n
54
Laws of Inheritance
Inheritable characters in human beings:

In human beings following characters are inherited:
1. Albinism: Whitening ( fully or partial) on skin, hairs or eyes is a
recessive character.
2. Blue colour and brown colour of eye balls: Blue colour of eye
ball is recessive character. brown colour is dominant character.
3. Inefficiency to taste thiocarbamide: Thiocarbamide taste bitter
but because of recessiveness human being is unable to taste it.
4. Baldness: Too much automatic removal of hair from head is a
dominant character.
5. Skin spotting: Premature whitening of hairs on front side of
head is a dominant character.
Numerica1s in genetics:
Exercise-1
Question 1. In sweetpea the purple flower colour results as an
interaction of two genes C and P, either of which alone produces
only white flower. What will be the flower colour in hybrids obtained
by following cross:
A. Ccpp x ccPp
B. Ccpp x CcPP
C. ccpp x CCpp
D. Ccpp x ccpp
E. CCPP x CcPp
F. CcPp x CcPp
Answer:
Summary
According to the question:
C= Independently produced white flowers.
P= Independently produced white flowers.
CP=On interaction produces purple flowers.
C=Produces white flowers.
I P=Produces white flowers.
(a) CcPp x ccPp
purple white
55
GtmI!tics : Fundamentals and Applications
In the first parent C and P on interaction produces purple flowers. In

the second parent P is single so it produces white flowers.The first
parent (CcPp) produces 4 types of gametes i.e. CP, Cp, cP, cp. The
second parent (ccPp) produces only two types of gametes i.e. cP and
cp. To form Fl generation these gametes will combine as mentioned
in following checkerboard:
CP Cp cP cp
cP CcPP CcPp ccPp ccPp

+ + o o
cp CcPp Ccpp ccPp ccpp

+ 0 =
+ 3 CP = Purple flowers : 3 purple flowers

1 Cp White flowers
0 3 cP = White flowers
= 1 cp White flowers
= : 5 white flowers
So, the phenotypic ratio is 3:5 because of following reasons:
+ there are two dominant genes.
there is one dominant gene.
= both genes are recessive.
(b) CcPp x CcPP
purple purple
First parent (CcPp) is purple because of interaction of two dominant
genes. Same thing is true with the other parent. First parent will
produce four types of gametes viz. CP,Cp,cP,cp. The other parent
(CcPP) produce two types of gametes viz. CP and cP. To make Fl
these gametes will combine as mentioned in following checkerboard:
56
Laws of Inheritance
CP Cp cP cp
CP CCPP CCPp CcPP CcPp

o o o o
cP CcPP CcPp ccPP ccPp

o o x X
•
o 6 CP = purple flowers
x 2 cP = white flowers
So phenotypic ratio is 6:2 or 3:1 because of following reasons:

o = two dominant genes.
x = it has one dominant gene.
(c) ccPp x CCpp
white white
Both the parents have white flowers because of presence of only one
dominant gene. First parent (ccPp) produce only two types of gametes
viz. cP and cp.Second parent ( CCpp) forms only one type of gamete
viz' Cp. To form F1 these gametes will combine as mentioned below:
cP cp
Cp CcPp Ccpp
o +
o 1 cP = purple flowers
+ 1 Cp white flowers
Hence, the phenotypic ratio is 1:1 because of following reasons:
o = 2 dominant genes.
+ = 1 dominant gene.
57
(d) Ccpp x ccPp

white white
Both parents have white flowers because of presence of only one
dominant gene. The first parent (Ccpp) will produce two types of
gametes viz. Cp and cp. The second parent (ccPP) will also form two
types of gametes cP and cp. To form F1 these gametes will combine
as demonstrated in following checkerboard:
Cp cp
cP CcPp ccPp
o +
cp Ccpp ccpp
o CP = purple flowered =1
+ cP = white flowered
= cp = white flowered
cp = white flowered =3
Hence, phenotypic ratio = 1:3 beca.use of following reasons :

o =2 dominant genes
+= 1 dominant gene
== 1 dominant gene
- - No gene is dominant
(e) CCPP x CcPp
purple purple
Both the parents have C and P two dominant genes therefore both
have purple flowers. First parent (CCPP) forms only one type of
gamete CP. Second parent forms four types of gametes i.e. CP, Cp,
cP, cp. To form F1 these gametes will combine as demonstrated below:
58
Laws of Inheritance
CP Cp cP cp

o o o o
o = 4 CP = coloured flowers because in every case two dominant

genes are present.
(f) CcPp x CcPp

purple purple
Both the parents have purple flowers because of presence of two
dominant genes C and P. The first parent (CcPp) will form four types
of gametes viz. CP, Cp, cP, cp. The other parent (CcPp) will also form
four types of gametes viz. CP, Cp, cP, cp. To form Fl these gametes
will combine as demonstrated in following checkerboard:
CP Cp cP cp

0 0 o o
Cp CCPp CCpp CcPp Ccpp

0 x o x
cP CcPP CcPp ccPP ccPp

0 0
cp CcPp Ccpp ccPp ccpp

0 x =
In other words,
o = 9CP = coloured flowers = 9
x = 3Cp = white flowers
59
3 cP = white flowers
= = 1 cp = white flowers = 7
Hence the phenotypic ratio = 9:7 because of following reasons :
o = two dominant genes
x = one dominant gene
- = one dominant gene
= = all recessive genes
Exercise 2
In summer squash plant gene ( factor) W expresses white colour of
fruits. It is also epistatic over Y factor which is responsible for yellow
colour of fruits. When only recessive genes (factors) are present (wy)
the fruit colour is green. What will be the genotype and phenotype of
the ofsprings ( progeny) of following crosses:
(i) wwYY x Wwyy
(ii) WwYy x WwYy
(iii) WwYy x wwyy
(iv) Wwyy x wwYy
(v) WWyy x wwYY
Answer:
Summary: As mentioned in the exercise -
W = Independently produces white. It is also epistatic over Y.
Y = In absence of W it produces yellow fruits.
WY = Produce white fruits.
wy = Produce green fruits.
(i) wwYY x Wwyy
yellow white
First parent ( wwYY) will form one type of gamete i.e. wY. The second
parent ( Wwyy) will form two types of gametes i.e. Wy and wy. To
form Fl these gametes will combine as demonstrated below:
60
Laws of Inheritance
Wy wy
wY WwYy wwYy
o +
Inother words,
o = 1 WY = white fruited
+ = 1 wwYy = yellow fruited
So, the phenotypic ratio = 1:1 because of following reasons:
0= W works as epistatic gene.
+ = Y is free to express yellow colour.

(ii) WwYy x WwYy
white fruits white fruits
First parent will form 4 types of gametes viz.WY,Wy, wY,wy.
Similarly the second parent will also 4 types of gametes such as WY,
Wy, wY,wy. To form F1 these gametes will fertilize as mentioned
below:
WY Wy wY wy
WY WWYY WWYy WwYY WwYy

0 0 0 0
Wy WWYy WWyy WwYy Wwyy

0 x 0 x
wY WwYY WwYy wwYY wwYy

.
0 0
wy WwYy Wwyy wwYy wwyy

0 x
61
In other words, 0 = 9 WY = white fruited plants

x = 3 Wy = white fruited plants = 12
- = 3 wY = yellow fruited plants = 3
= = 1 wy = green fruited plants = 1
Hence, phenotypic ratio is 12:3:1 because of following reasons:
o = W is epistatic to Y
x = W is free to act
- = Y is free to act
= = wy both are recessive genes
(iii) WwYy x wwyy
white green
First parent (WwYy) forms four types of gametes viz. WY, Wy,
wY,wy.Second parent (wwyy) will form only one type of gamete
i.e.wy. To form Fl these gametes will fertilize as indicated below:
WY Wy wY wy
wy WwYy Wwyy wwYy wwyy

o x +
In other words, 0 = 1 WY = white fruits

x = 1 Wy = white fruits = 2
- = 1 wY = yellow fruits = 1
+ =1 wy = green fruits = 1
Hence, phenotypic ratio = 2:1:1 because of following:
o = W is working as epistatic gene
x = W is free in action
Y is free in action
- - wy both are recessive genes
(iv) Wwyy x wwYy
white yellow
The first parent will form 2 types of gametes i.e. Wy and wy . The
other parent will form two types of gametes wY and wy. TO form Fl
these gametes will fertilize as mentioned below:
62
Laws of Inheritance
Wy wy
wy WwYy wwYy
o
wy Wwyy wwyy
+ x
In other words, 0 = 1 WY = white fruit

+ = 1 Wy = white fruit = 2
- = 1 wY = yellow fruit = 1
x = 1 wy = green fruit = 1
So the phenotypic ratio is 2:1:1 because of following reasons:
o = W is acting as epistatic gene.
+ = W is free to act.
- = Y is free to act.
x = wy both genes are recessive.
(v) WWyy x wwYY
white yellow
First parent ( WWyy ) will form one type of gamete i.e. Wy. Other
parent will form one type of gamete i.e. wY. To form F1 the gametes
will fertilize as given below:
Wy
wY WwYy
Because in every case W is acting as epistatic over Y. Therefore all the

F1 generation will express white fruits.,
Exercise 3
A purple flowered variety in summersquash plant gives rise to 3/8
purple and 5/8 white in Fl when crossed with another white flower
variety. Find out genotype of both the parents.
63
Answer:
Suppose A and B genes (factors) independently produce white
fruits but when these genes come together then produce purple fruits
due to gene interaction. In that case genotype of both the parents
will be:
Purple parent = AaBb
White parent = Aabb or aa Bb
It can be proved by following checkerboard. Purple parent will form
4 gametes viz. AB, Ab, aB ,ab. White parent will form Ab, ab or aB,
ab. Various gametes will combine in following ways to form Fl.
(a) AB Ab aB ab
Ab AABb AAbb AaBb Aabb

o o
ab AaBb Aabb aaBb aabb

o + x
In other words, o = 3 AB = purple because both are dominant

genes.
- = 3 Ab = white because A is free in action.
+ = 1 aB = white because B is free in action.
x = 1 ab = white becallse both genes are recessive.
So the phenotypic ratio is 5 white : 3 purple
(b) AB Ab aB ab
________________________________________ ab
AaBb Aabb aaBb aabb
o + x
aB AaBB AaBb aaBB aaBb

o o + +
64
Laws of Inheritance
In other words, 0 = 3 AB = purple because both are dominant

genes.
- = 1 Ab = white because A is alone.
+ = 3 aB = white because B is alone.
x = 1 ab = white botlt are recessive genes.
Therefore phenotypic ratio is 3 purple: 5 white.
Hence, genotype of purple = AaBb
Genotype of white = Aabb or aaBb
Questions for practice
(1) In following crosses, in which genotype of parents are given,
what are the gametes produced by each parent and what will
be the flower colour of the offsprings from each cross : Rr x RR,
rr x Rr, RR x rr, Rr x Rr ?
(2) What will be the appearance of the offsprings of following crosses
in which genotypes of parents are given:
(a) TTGgRr x ttGgrr
(b) TtGGRr x TtGgRr
(c) ttggRr x TtGgrr
(d) Ttggrr x TtGgRr
(3) If the homozygous dwarf, green and wrinkled pea plant is
crossed with a homozygous tall, yellow and round one, what
will be the appearance of the F1 ? What gametes does the F1
form ? What is the appearance of the offspring of a cross of the
F1 with its dwarf green & wrinkled parent and with its tall ,
yellow and round parent?
(4) In snapdragons tall (T) is dominant to dwarf (t) and red flower
(R) are incompletely dominant to white (r), the hybrid being
pink. A pure tall white is crossed to a pure dwarf red. The F1 is
self fertilized. Give the F2 phenotypic ratio.
(5) In 4 oclock plant ( Mirabilis jaZapa ) a plant which is heterozygous
for red flower (R) and white (r) is pink. What will be the flower
colour of the F1 produced and in the F2?
(6) Purple flowered x white flowered gives 50 per cent purple
and 50 per cent white. Give the genotypes of the parents.
(7) A green maize plant when selfed produces about 15/16 green
and 1/16 white (lethal) seedlings. Find out the genotype of the
plant and the mode of inheritance of the pigment.
65
(8) Wheat plant has 21 pairs of chromosomes. State the number of

chromosomes present in'the (a) stem tip (b) nucleus (c) pollen
type nucleus (d) embryosac (e) seed embryo (f) endosperm.
(9) In fowl the dominant gene R gives rosecomb and the dominant
gene P gives peacomb.When P and R are present together the
comb form is walnut. The homozygous recessive of P and R
produce single comb. Determine the comb form of the offspring
of following crosses.
a. RrPp x RrPp
b. rrPP x RrPp
c. rrPp x RRPp
d. Rrpp x RrPp
e. Rrpp x Rrpp
f. rrPp x Rrpp
(10) Both parents of a blue eyed man are brown eyed. He marries a
brown eyed women who had a brown eyed mother, a blue eyed
father and a blue eyed brother. The man and women in question
have a brown eyed child. Give the genotypes of (i) parents of
man and women (ii) man and women (iii) their child.
'66
Chapter 7
Heterosis or Hybrid Vigor
It is a general observation that when dissimilar varieties of plants
and animals are crossed then the F1 hybrid is more robust as compared
to the parents (Fig-15). This quality of hybrid is called hybrid vigor
or heterosis. It can express in several ways e.g. increased size,
resistance to diseases or pests, easy propagation by vegetative means,
more yield etc.
First report of hybrid vigor was available from Kolereuter in 1763
while experimenting on tobacco. Afterwards several scientists
reported this phenomina in several plants and animals. A good example
in animal is mule which is a hybrid between crossing of ass with
mare.
A special feature of hybrid vigor is that it get reduced in F2 and
further reduced in future .generations.
Hybrid
Fig. 15: Hybrid vigour in maize. Outer plants are parents of the middle
plants.
67
Occurrence of hybrid vigour ( heterosis) in plants and animals

Hybrid vigour ( heterosis ) has been reported in several plants and
animals. Such plants are maize, jowar, barly, pearl millet, cucumber,
onion, brinjal, tomato, forage crops, sunflower, sugarcane, coconut,
tobacco, penicillium fungus etc. Examples in animals are mule, rats,
cattles, silkworms etc.
Causes of heterosis (hybrid vigour)
Several scientists from time to time have proposed several causes.
But none has been found to be fully satisfactory. Various causes
have been explained as below:
(1) Heterozygosity induces physiological stimulus:
This idea was proposed by East and Hays in 1912. They expressed
that hybrid vigour is due to heterozygosity in hybrids. Union of
dissimilar genes induces physiological stimulations which lead to
hybrid vigour.
(2) Greater initial capital hypothesis:
This hypothesis was proposed by Ash in 1930. According to him in
such hybrids there is increase in embryo size and capacity which leads
to hybrid vigour.
(3) Dominant gene (factor) hypothesis:
The scientist Bruce proposed this hypothesis.. According to him number
of dominant genes is more in hybrids compared to their parents.
This fact results hybrid vigour.
(4) Linked favorable dominant gene hypothesis:
This hypothesis was proposed by Jones in 1917. According to him
from the parents growth favorable linked dominant genes come in
the hybrid which causes hybrid vigor.
(5) Cytoplasmic nuclear hypothesis:
This hypothesis was proposed by the scientists Michaelis, Shull and
Lewis. They expressed that hybrid vigor is an induction due to
interaction between nucleus and cytoplasm.
68
Heterosis or Hybrid Vigor
Effects of heterosis
A. Morphological effects~
(1) Root: Increase in diameter, length and number of branches.
(2) Stem: Increase in thickness, wrinkleless, length and number
of branches.
(3) Leaves: Increase in number, size and intensity of green col or .
(4) Flowers: Increase in number, size and beauty.
(5) Fruit: Increase in size and quality.
B. Physiological effects:
1) Increase in germination.
2) Increase in cell diT/ision and cell size.
3) Early flowering.
4) Resistance to drought, diseases and pests.
5) Increased life.
6) In animals increase in tolerance and working capacity.
Role of heterosis in increasing production:
Productivity is the power of producing or multiplying which is the
result of several biological and environmental factors accumulated as
effective stable system. In agricultural context productivity is yield.
Yield is a complex parameter reflecting the influence of a number of
systems on the final product. The influences can be identified as seed,
soil, water, climate, management etc. These can further be subdivided
into several components. For example the seed factor can be a good
high yielding genotype or a local nondescriptive genotype. Among
high yielding genotype it can be an improved variety or synthetic or
composit or a F1 hybrid seed which is expected to be heterozygous
for most of loci. Higher production than parents is known as heterosis
Commercial F1 hybrids available in India are limited to maize ,
sorghum, bajara, cotton etc in which substantial output through
heterosis is being utilised for increasing food and fibre production.
In next order comes the crops like castor, arhar, melons, mango etc.
where heterosis was identified and exploited on limited scale.
In crops like rice, wheat & pulses substantial success was achieved
through conventional breeding, polyploidy, mutation breeding etc.
in developing superior genotypes. Extensive use of dwarfing genes
(discovered by Vogal in USA & Borlaug in Mexico) particularly in
69
wheat has brought out redistribution of dry matter for higher harvest
index and resistance to lodging under high fertility conditions.Indian
wheat varieties have harvest index of upto 49 per cent. In case of rice
the dwarfing gene discovered by Taiwan scientists lead to
restructuring of rice types like IR 8 to IR 26 Apart from these Indian
scientists have developed several varieties for high productivity that
are suitable to different regions. Sorghum, maize, bajara etc. occupy
a large part of cultivated area. Those are main food crops of dry land
farmers. The basic scientific approach for improvement of these crops
has been heterosis breeding. India's high yielding hybrids of sorghum
and pearlmillet are considered one of the best in world.
New arhar varieties of less than 5 months duration can be grown in
rotation with wheat crop. The short duration varieties of moong
maturing in 60 -70 days can be planted during spring and summer
months when normaly the land is kept fallow.
Achievements by heterosis in various crops:
(a) Sorghum hybrid varieties: CSH-l to CSH-I0
(b) Pearlmillet hybrid varieties: HB-l to HB-6
BJ-104 and BK-559 are resistant to downy mildeu
(c) Maize hybrid varieties: DHM-101 & 103 and Histarch.
(d) Cotton hybrid varieties: H-4, Varalaxmi, DCH-32, JKH-l
etc.
(e) Castor hybrid varieties: CCH-l, CCH-3 etc.
(f) Sunflower hybrid varieties: BSH-l,
(g) Groundnut hybrid varieties: J-24, Kadiri-3
(h) Mango hybrjd varieties: Amrapali and
(i) Cattle hybrid varieties:
Sahiwal x Holstein - Friesian
Ongole x Jersey
Haryana Zebu x Brown Swiss
Effect of breeding on increase of milk yield has been estimated to be
38.7% apart from the effect of feeding and management (Ram and
Chowdhary,1984)
70
Chapter 8
Mutation
Usually genes get distributed without any change from one cell to
another by mitosis or from one generation to other generation by
gemetes thorough meiosis. But sometimes due to some special
circumstances genes get chemical change due to which new varieties
develop. Such changes may also happen in structure of chromosomes
and sequence of genes. Such changes in gene or chromosome are called
mutation. Changes in gene are called gene mutation. Changes in
chromosome may be called chromosome mutation. Genetic or
chromosomal changes in somatic cell are called somatic mutation. The
mutations in germinal cells (gametes) are important in evolution.
These mutations develop new varieties of organism.
Definitions of mutation:
Sudden changes in hereditary material are mutation (Poelman). It
may happen is following ways:
(i) change in constitution of gene.
(ii) Change in sequence of genes of chromosome.
(iii) Duplication or loss of segment of chromosome.
Types of mutation:
Mutation may be of two types viz.
a) Spontaneous mutation: It is developed by nature
b) Induced mutation: These are produced by physical or chemical
mutagens.
According to nature of tissues mutation may be two types.
a) Somatic mutation: It occurs in somatic cells so it may not be
heritable.
b) Germinal mutation: This mutation happens in germ cells
(gonads, anthers and ovary or ovum). It is of following types:
(i) Biochemical mutation: These mutation cause chemical or
physiological effect.
71
(ii) Spurious mutation: Due to this mutation recessive character

gets expression.
(iii) Point or gene mutation: It is caused by change is structure
of gene.
(iv) Chromosomal or anamozygous mutation: This mutation
causes change in chromosome number or change in
arrangement in segment or gene. This mutation can be of
following types.
Mutation leading to change in number of chromosome:
It is two types:
(1) Genomatic mutation: Reduction in number of chromosome pairs
may be haploidy. Increase in numbers of chromosome pair may
be polyploidy
Heteroploidy i.e. increase or decrease in number of chromosome in
chromosome pair.
Mutation leading to change in arrangement of segment or gene. It is
of following types.
(i) Intrachromosomal segmental rearrangement: It is also called
chromosomal inversion.
(ii) Inter chromosomal segmental rearrangement: It is also called
translocation.
(iii) Delection or duplication : It is either decrease in chromosome
segment or gene (deletion) or increase in chromosome segment
or gene (duplication).
Nowadays the world mutation is used only for gene or point
mutation only. Mutation may be dominant or recessive. Dominant
mutation is expressed in Fl generation. However recessive mutation
may not be expressed for several generations unless it attains
homozygosity .
Stages where mutations may occur
Through mutation may happen at any stage or life. But its occurrence
is more likely in following conditions:
(i) Mutation may happen in mature gamete. So where this gamete
goes only that offspring gets this mutation.
(ii) Mutation may occur before meiosis in primary gametocyte.
Therefore half of the gametes which are produced from this
gametocyte develop mutation.
72
Mutation
-(iii) After fertilization, at anaphase of mitosis daughter chromatids

form, Mutation may occur in one of the daughter chromatid. So
it may effect half of the body.
(iv) It item 3 both side chromatid get mutation then the whole body
may get affected.
(v) Mutation may occur in sex chromosomes X or Y.
(vi) So either male or female generation gets affected .
. (vii) During growth of the body mutation may occur only in one cell.
So from that mutated cell the developed organ may get affected.
Size of mutation:
Size of mutation is studied from difference in parent and mutated
gene. If the difference is clear then such mutation is called prominent
or conspicuous mutation. If the difference is vary small then such
mutation may be called as inconspicuous mutation
Frequency of Mutation:·
Possibility of frequency of mutation may be less in a gene. But plant
or organism are made of million of cells. Therefore the chance increases
for more frequency of mutation. Some scientists have found out
frequency of mutation in some organisms as mentioned below.
1. In Drosophila melanogaster fly mutation frequency in x sex
chromosomes is 0.15 percent per generation. In the second
chromosome it is 0.5 percent and in the third chromosome the
mutation frequency is 1.2 percent
2. In maize (Zea mays) in R gene the frequency of mutation is 492
per million. In C gene the frequency is 105 per million. In the
gene Pr the mutation frequency is 11 per million
3. In human beings out of 24 thousand gametes only in one gamete
mutation frequently has been found out.
Effect of mutation:
Mutation maybe harmful or usefull or lethal in effect. Most of the
mutations have harmfulleffect. Some times mutation have useful effect
for example seedless grape, naval oranges and disease resistant
varieties of maize and wheat.
Due to harmful effect of lethal mutation developing embyo are killed.
Lethal mutation may appear in autosomes and sex chromosome. In
maize because of lethal mutation development of chlorophyll stops
due to which the seedling gets killed.
73
Genetics: Fundamentals and Applicatiolts
Properties of mutation:
Mutation has following special properties:
(1) Not gradually but suddenly mutations appear.
(2) Mutation is heritable.
(3) Mutation appear in one or very few organism.
(4) Effect of mutation may be morphological, physiological or
biochemical. In Drosophila melanogaster due to mutations antennae
changes into legs, balancers converts in second pair of wings
and sucking mouth parts are changed to mouth parts of inferior
quality.In Oenothera plant mutation has developed several
varieties which have different types of sizes, leaves and flowers.
(5) All mutants either developed from prominent or inconspiquous
mutations match more or less with ancestors.
Mutagen:
Mutagen means the device to induce mutation. Mutagens are of two
types i.e. physical mutagen and chemical mutagen. Muller was first
to find out physical mutagen. In 1927 Muller irradiated Drosophila
melanogaster with X ray which produced mutation. Since then scientists
are using high energy penetrating ionizing radiations for example x
rays, gama rays, alfa rays, beta rays, neutron, cosmic rays, radioactive
isotopes, ultraviolet rays etc. as physical mutagen to produce mutation
in various plants and anim~ls.
In addition to physical mutagens several chemical mutagen are also
being used such as mustard gas, ethyl methane sulphonate, ethyl
urethane, magnous salt, high temperature, formaldehyde, eosin,
enthrosine etc.
Method to induce mutation:
For induction of mutation plants, seeds, male & female gametes or
whole body are trE!ated with suitable doses of physical or chemical
mutagen. After treatment with mutagen M1,M2,M3,M4, etc.
generations are grown. Selection for desirable character is made in
each generation. After a variety is developed it is subjected to field
testing in replicated trials. Afterwards the improved variety is finaly
released.
74
Mutation
Problems of induction of mutation in fruit trees and ways to tackle

a) Problem of diplontic or intra-somatic selection in irradiated
tissue which necessitates an effective screening technique
for recovering wholly mutated tissue from an initially
induced chimeric situation.
b) Long juvenile phase of fruit trees rendering the evaluation
of radiation effects, a delayed and long range programme.
c) The question of land space & technical know how for proper
evaluation of large number of population.
d) Primary and secondary ploidy nature of fruit trees limits
the chance of detectable mutation, though total mutational
events might have been more.
e) Threshold value of mutagenic and lethal dose is
comparatively narrow with acute radiation.
Nakajima (1965) from Japan reported a gradual cutting back technique
towards the lower buds to recover a large number of somatic mutants
in chronically irradiated roses. Das (1969) by nipping off the
undesirable buds and forcing up buds only from charged axil could
recover totally mutated twigs in irradiated Hibiscus.
Biological effect of irradiation on mutation:
Radiations are of two types -long waves (wave length 2000 to 2950
o A) and short waves (wave length 0.06 to 1000 0 A). Long wave rays
(e.g. ultra violet radiation) do not express ionization and have less
capacity for penetration. But short wave rays such as x rays and gamma
rays (wave length 9.01 to 1.4 0 A) express ionization and have more
capacity for penetration. Both of these radiations can damage cells.
These radiations are used for disinfection or for induction of mutation.
In case of short wave radiations very less dose is used for mutation
induction in plants and animals which are used for human welfare.
To explain ionizing radiation geneticist have given following theories:
(1) Target theory: According to this theory the ionizing radiat
energy hits certain targets such as enzyme molecule, gene
molecule, nucleotide or muton. Its use in increased dose causes
more effects to the celL
(2) Free radical theory: This theory explainS that with the effect of
radiation nonspecific material or water gets broken down due
to which many free radicals such as H2 02 are formed. After
some reactions it forms organic peroxidase. These cause several
75
oxidation - reduction reactions resulting changes in

chromosomes.
Ionizing radiations are more sensitive to nucleic acid (DNA and RNA)
as compared to cytoplasmic structure (example enzyme) . Ionizing
radiation reduces cell division. In addition to it chromosomes also
gets affected. Chromosomes get breakage or there happen change in
genes. In this way all this results genetic mutations. Its genetic effect
is irreversible and permanent.
Certain organisms are sensitive to high temperature treatment for
.development of mutations. In contrast to it, certain organism responds
to low temperature treatment for production of mutants. We can take
the example of human eye disease Aniridia. In eyes, for development
of iris only one gene is responsible. Due to certain reasons this gene
gets changed (mutated) and become unable to form iris. If this gene
present in gonad, gets mutated then it gets transferred to zygote.
Through mitotic cell division this mutated gene reaches to every cell
of the body. But it expresses its effect at proper time only. Due to this
mutated gene, development of iris do not occur which causes the
disease Aniridea. It is due to the dominant mutation. The parent has
this mutated gene in his/ her gonads. At sexual maturity half of the
gametes get this mutated gene. Due to which half of the offsprings
inherit this eye disease Aniridea
Methods to find out mutation:
(A) Mutation in sex chromosome: To find out mutation muller has
developed a good method knwn as CIB Method. This method suits
to those organisms in whose x chromosomes mutations are produced
artificially. In this method such female Drosophila melanogaster in whose
x chromosomes there is a cross over suppressor gene C, a recessive
lethal gene I and a dominant gene B responsible for Bar eye, that
female Drosophila fly is irradiated with x ray or other suitable
mutagen and then hybridized with male Drosophila melanogaster. In
Fl Bar eyed daughters CIB chmmatid comes from mother and x
chromatid comes from father. The Bar eyed daughter fly was crossed
with normal male. In F2 all such sons died which received CIB
chromatids from mother. In other sons recieving x chromatid, if those
x chromatid get lethal mutation then those sons also die. But in F2 all
the daughters were alive because those received normal x chromatid
from father ( Fig-16)
Hence the above experiment reveal that in Fl the CIB female produce
76
Mutation
, X~rays
c B
__,--' cl X-9 ..
F,
c s+ B .+... 1
+. . .+r
C
........
~.,
8 ... + +
... ... I + J
\,
I ,.
..'
C e i I
Ft • ... ... 1...
... ... . di8I
Fig. 16: Diagram showing the CIB method of detecting lethal mutation.
only daughters and no son at all. This means that lethal gene from
father came to sons and so all sons died. Now to know that whether
that lethal gene was developed by itself or by treatment we have to
perform two experiment together.
In one experiment mutagen treated male was hybridized with CIB
female of Fl. In other experiment normal male was hybridized with
CIB female of Fl. If in the experiment with mutagen treated male
most of the offsprings died because of lethal gene then one conclusion
could be drawn that the lethal gene was developed by mutation
treatment.
Second conclusion is that if in the mutagen treated x chromosome
there is any prominent (visible) mutation then that appear in the sons
of F2 generation.
(B) Mutation in autosomes:
CyLpm method of finding out mutation is popular for autosomes. In
Drosophila melanogaster fly second chromosome's one sister chromatid
had gene Cy for curled wings and gene L for lobed eye. The second
chromatid had gene pm for brown colour. The three genes (Cy, L,
pm) are lethal if present in homozygous condition.
Drosophila melanogaster fly with Cy L pm genome is crossed with x ray
irradiated Drosophila fly. The result is that in Fl generation the gene
77
Cy, L or pm comes from one parent and x ray treated chromatid

comes from other parent.
Now from F1 any type of fly which had either (a) Cy, L and x ray
treated chromatid, (b) had the gene pm and x ray treated chromatid.
Such fly was crossed with similar type of Drosophila fly. In the F2
three types of offsprings appear in 1:2:1 ratio given below:
1 = Cy L/ Cy L or pm / pm double chromatid
2 = Double treated chromatid
3 = One chromatid with Cy L or pm genes and the other x ray treated
chromatid.
In this experiment the first type of Drosophila dies. Third type of
Drosophila survive. Second type of Drosophila either survive, but dies
only if lethal mutation appeared in the treated x chromosome.
Importance of mutation in evolution:
Evolution of any organism is possible only by inherent variations
developed by mutation. Mutation just appears not for any purpose.
If it can be useful then that can be helpful in evolution. Generally
inconspicuous mutations are more useful for evolution. Importance
of mutation is given below:
(a) Importance of gene mutation in evolution:
For evolution gene mutations are comparatively more useful. But
Deveries was of the opinion that only conspicuous mutations are useful
in evolution. However on further researches this opinion of Deveries
was found baseless. Inconspicuous mutations accumulate to develop
sufficient evolutionary changes. After Watson and Crick explained
the structure of gene since then scientists have the opinion that
mutation happen because of changes in genetic code.
(b) Importance of chromosomal mutation in evolution:
Under this duplication, inversion and translocation have much
importance. Deficiency is not important because it is lethal for
organisms. Duplication results increase in number of genes resulting
new characters, so it is of importance in evolution. Translocation and
inversion generally bring about varietal difference and species
differentiation. For example, two varieties of Drosophila melanogaster
differ because of inverted segment. In Datura stramonium varieties
have developed spontaneously due to occurrence of reciprocal
translocation.
78
Chapter 9
Polyploidy
If more than two homologous chromosomes sets are found in cell or
organism then that is called polyploid. Polyploidy is more found in
plants as compared to animals. The reason for less polyploidy in
arrlmals is their sex balance. In plants sex balance is weak. In animals
if number of chromosome becomes more than 2n they become intersex
and loose the capacity for reproduction. So it is very rare in animals
such as in Artemia salina.
Origin of polyploidy:
Polyploids are produced in several ways. Some time meiosis happens
in anamolous way due to which number of chromosome is not halved
but the number remain unchanged and it forms gametes. Two such
gametes fertilize and form tetraploid.
During mitotic cell division at anaphase chromatids may not move to
the poles. Another situation may happen where cell wall does not
form. Results of both these situations are somatic doubling i.e.
duplication of the somatic chromosome number. In other words
formation of a tetraploid cell takes place.
On fertilization of a tetraploid plant with a diploid plant birth of a
triploid plant takes place. Somatic doubling of a triploid will lead to
development of hexaploid plant. On fertilization of a hexaploid plant
with a tetraploid plant, a pentaploid develops.
Types of polyploidy:
Differences in chromosome number results two types of polyploidy
i.e. (a) Euploidy and (b) Aneuploidy.
(a) Euploid: Euploid is the exact multiplication of chromosome set.
Euploid is also known as true polyploidy. Euploid is of following
two types:
(i) Autopolyploid or Autoploid
(ii) Allopolyploid or Alloploid
79
1. Autopolyploid or Autoploid:
If chromosome number of diploid due to any circumstances becomes
double then polyploidy formed in this way is called autopolyploid or
autoploid (Fig- 17). For example AA genome if becomes AAAA then
it is autotetraploid. For development of new varieties or species
autopolyploidy has great role.
AAAA
T
Fig. 17: Origin of auto and allopolyploids.
2. Allopolyploid
On hybridization of different diploid species, hybrid get developed.
If somehow chromosome number of hybrid gets doubled. Then the
polyploidy developed in this way is known as allopolyploid or
alloploid. This is important for development of varieties and species.
For example, a diploid species with AA genome gets hybridized with
a different BB diploid. Then the hybrid will have AB genome. This
Raph.nus x Brassk:8 :0 Rapbanoblalalc8
Raclllh Cabbage Rabblge
(2n "18) (2n "18) (2n -18)
RRRRRRRRR CCCCCCCCC RRRRRRRRR
RRRRRRRRR CCCCCCCCC CCCCCCCCC
Fertile F8I1I1e SterIle
(synap~ falluret
(2n) Rabbage ColChicine... Tetraploid (411) Rtbbage

RRRRRRRRR RRRRRRRRR CCCCCCCCC
CCCCCCCCC RRRRRRRRR CCCCCCCCC
The FonnatIon Of A Fertile Tetraploid Rabbage
R .. radish chronIosome C - cabbage cIIromotome
Fig.1S: Cross of radish (Raphanus) with cabbage (Brassiea) leading to

formation of allopolyploid Raphno brassiea .
80
Polyploidy
genome is sterile to some extent. On duplication of its chromosome

number the tetraploid formed is known as allotetraploid. The sterility
disappears by the allotetraploidy. Such allopolyploid are known as
amphidiploid. It is fertile like diploid. Some examples of amphidiploid
are Primula kewensis, Crepis rosea, Iris versicolar, Serpentina townsendii
and Raphno brassica etc.( Fig-18).
(b) Aneuploidy:
Sometimes in nucleus there is no exact multiplication of chromosome
set such as 2x + n or 2x- n. Such a situation of polyploidy is known as
aneuploidy. Due to aneuploidy the genic balance gets disturbed
leading to sterility.
Aneuploidy is of several kinds for example monosomic ( 2x-l) example
tobacco and Drosophila melanogaster ; trisomic ( 2x+ I) example datura,
maize & tomato; tetrasomic ( 2x+2) example wheat and double
trisomic (2x+I+I) example maize.
Aneuploid if increases the number of chromosome for example 2x+1
then this situation is known as hyperploid. If aneuploid bring down
the number of chromosome then that phenomenon is called hypoploid.
Aneuploidy has great role in evolution.
Polyploid series:
While study of various genus of plants it has been observed that the
basic chromosome number get multiplied in form of series. This type
of series is called polyploidy series. Some examples of polyploidy
series are given in the following table:
Plant! Polyploid Types of polyploidy basic
Genus series
number
1 Triticum wheat 14,28,42 Diploid, Tetraploid, Hexaploid 7
2 Gryza Rice 24,28 Diploid, Tetraploid 12
3 Rosa Rose 14, 21, 28, 35, Diploid, Triploid, Tetraploid, 7
42,56 Pentaploid, Hezaploid,
Octaploid
4 Solanum Potato 12, 24, 36, 48, Diploid, Tetraploid, Hexaploid, 6
60, 72, 108, Octaploid, Decaploid, 12 ploid,
144 18 ploid, 24 ploid
5 Psidium Guava 22,33,44,55 Diploid, Triploid, Tetraploid, 11
Pentaploid
81
Genetics: Fundamentals and Applicatiolls
Effect of polyploidy
Polyploid organisms (mostly plants) as compared to normal organism
are different. This difference is morphological or physiological or
genetical as described below.
(A) Morphological effect of polyploidy:
1. Due to polyploidy stem become thicker and stouter.
2. Polyloid leaves are larger, thicker and deeper green.
3. Polyploid leaf hairs are thicker and longer.
4. Polyploid plants are of larger size.
5. Polyploid flowers are larger. Its sepals, petals, stamens and
ovary become large and attractive.
6. Pollen of polyploid plants are larger.
7. Generally due to polyploidy fruit size and quality increases. So
their economic value increases.
8. Sometimes due to polyploidy fruits become irregular so their
economic value gets reduced. Example triploid guava.
9. Cotyledon size increases in polyploidy.
10. Polyploidy reduces reproductive power. The reason is that due
to multivalent formation at anaphase distribution of
chromosomes is not proper. This leads to genetic unbalance
resulting into sterility.
11. In polyploids number of flower gets reduced.
12. Due to polyploidy cell size of stomata and xylem increases.
13. Polyploidy increases size of cell and nucleus.
14. Polyploidy increases resistance to diseases.
(B) Physiological effects of polyploidy:
1. Generally polyploidy reduces growth rate.
2. In certain cases polyploidy changes annual habit to perennial
habit, example autotetraploid maize.
3. Polyploidy delays flowering.
4. In certain plants e.g. tobacco polyploidy increases the content
of alkaloid nicotin.
5. Polyploidy increases ascorbic acid content in certain plants such
as tomato, cabage, guava etc.
6. Due to polyploidy in tobacco ( Nicotiana tobacum) there is less
82
Polyploidy
carbohydrate and sulphure but there is increase in content of

nitrogen, calcium, potassium and magnesium.
7. Polyploidy enhances vitamin content.
8. Polyploidy increases osmatic concentration.
Polyploidy increases water content in plants.
(C) Genetical effect of polyploidy:
1. Polyploidy increases the content of allelomorphic genes.
2. Compared to diploid the autopolyploids are more stable
genetically.
3. Polyploidy increases the rate of mutation.
Artificial production of polyploidy:
Because of economic importance of polyploids and looking its breeding
possibilities following techniques may be tried to produce chromosome
doubling at somatic cell division of zygote. To achieve it ionizing
radiation, non ionizing radiation, temperature etc. can be tried.
Doubling of chromosome number in somatic cell:
It may be by following treatments -
(a) Irradiation with ionizing radiation: Irradiation with x ray at
seedling stage mostly result indoubling of chromosome number.
Other radiations such as gamma rays also can be tried.
(b) Heat treatment: Treatment of roots with high or low temperature
are used for chromosome doubling.
(c) Treatment by callus formation: On decapitation of stem or
branches, afterwards callus formation takes place. One of this
callus cells may happen to be polyploidy. It may grow a polyploid
branch. By this technique Winkler produced tetraploid in Solanum
nigrum.
(d) By chemical treatment: Several chemicals may be tried to induce
polyploidy. Some examples are colchicine, chloroform, alchohol,
lactic acid, chloral hydrate, nichotine sulphate and
phytoharmones e.g. alfa NAA, beta NAA, gibberelic acid, beta
IAA and IBA etc.
(e) Hybridization: New polyploids are formed by hybridization
between various ploidy levels.
83
Use o-f abnormal meiosis for production of polyploidy:

For production of polyploidy to induce abnormal meiosis ionizing
radiations, chemicals and hybridization techniques are used. Above
mentioned methods may be used for induction of polyploidy.
However colchicine is most commonly used.
Colchicine treatment for polyploidy induction:
Nowadays colchicine treatment is most popular technique to induce
polyploidy. Colchicine is an alkaloid. Colchicine is extracted from
root and corm of two plants of Liliaceae family- Colchicum autumnale
and Colchicum luteum . Indian scientist Parthasarthy reported
colchicines from the plant gloriosa superba of the family Liliaceae. Some
scientists from USSR haye reported that a chemical Granoson is also
effective like colchicine for polyploidy induction.
For use of colchicines there are several techniques used in plants. It
should be used only when the mitotic cell division is maximum in the
plant. The techniques are given below:
(a) Young and developing twigs are immersed in weak solution of
colchicine for suitable time, at time of maximum mitotic cell
division.
(b) Weak solution of colchicine may be dropped slowly for sometime
at maximum mitosis.
(c) Spray the weak solution of colchicines at timely intervals on
growing twigs at times of maximum mitosis.
(d) Injection of weak solution of colchicine in growing twigs at times
of maximum mitosis.
(e) Seed treatment with colchicines for suitable timings, washed and
then sown.
(f) In agar or lanoline paste colchicine in suitable concentration is
mixed and applied on growing tissue of plant to induce
polyploidy.
The techniques of colchicines application for induction of polyploidy
was invented by Blackslee, Avery and Nebel in 1937. Afterwards
several scientists such as Derman ( 1940, 1953),Luong (1951), Pope
and Love (1952), Brewbacker (1952), Hutton ( 1953 ), Hunter (1954)
and Evans (1955) have induced polyploidy by colchicine. Several
Indian scientists such as A.K. Sharma, R.P. Roy, A. Abraham, M.S.
Swaminathan and many others have developed several useful
polyploids in several crops such as cotton, sugarcane, gram, tobacco.
rye, rice, barley, oat, wheat, oil seeds etc.
84
Polyploidy
Importance of polyploidy in evolution:

Maximum amount of polyploidy have been reported in angiosperm
and perennial herbs by Stebbins in 1938. Medium amount of polyploids
are found in animals. Least amount of polyploids are found in woody
plants. Muntzing (1936) suggested that polyploidy may change annual
plant into a perennial plant.
From point of view of evolution polyploidy is important. In evolution
of polyploidy a time comes when it behaye like diploid. Polyploids
adjust to changed circumstances by modification in physiological,
morphological and genetical characters. This fact is important in
evolution.
By synthesis of allopolyploids new genera and species may get
Jormed. It may play great role in evolution. Karpechenko (1928)
hybridized radish (Raphanus sativus) with cabbage (Brassica oleracea).
These plants belong to different genus but have same number of
chromosomes. The hybrid had 18 chromosomes but these
chromosomes from father and mother were non homologous. So
during they failed to pair. Therefore they produced sterile gametes.
Due to some reasons meiotic cell division failed so the number of
chromosomes could not be reduced - it remained 18 only. In due
course of time such male and female having 18 chromosomes could
fertilize and produce a hybrid with 18 chromosomes. On doubling
this 18 chromosome number a allotetraploid with 36 chromosomes
was formed . It was fully fertile because of full pairing of
chromosomes. This was named as Raphano brassica by Karpechenko.
Compared to parents this hybrid was larger. Its pods were larger
and had more seeds.
Role of polyploidy in plant breeding:
Objective of plant breeding is to develop superior varieties with more
yield, disease resistance, better quality etc. Generally polyploids have
most of these features.
But in certain cases polyploidy has negative effect such as less yield,
poor quality, small size, slow growth and lower fertility. This fact
means that there is a saturation point beyond which increase in ploidy
brings about negative effect. Similar ideas were presented by Wettstein
(1927), Levan (1942), Stebbins (1952) etc. Therefore there is an
optimum level of polyploidy for every type of plant. It is called
optimum level of polyploidy.
85
For improvement of plant, hybridization is a good technique. But in

certain cases FI hybrids are sterile. To make these fertile polyploidy
breeding is a boon.
In plant breeding as compared to autopolyploidy the allopolyploidy
is more helpful because in autopolyploids due to more multivalent
formation the genic balance is disturbed. So they are sterile.
86
Chapter 10
Linkage
In the chapter on Laws of Inheritance it has been explained that there
are some exceptions to Gregor Mendel's law of independent
assortment. In F2 segregation ratio of characters is not always 9:3:3:1.
The differences are more in case of trihybrid and polyhybrid crosses.
These types of exceptions were first reported by Batson in England
and Morgan in USA.
Law of segregation and law of independent assortment are infact
based on locations of various pairs of segregating genes on non
homologous chromosomes. Hence larger the size of chromosomes
number of genes are there. Therefore at times of independent
assortment more combinations will be expected. Keeping these facts
in mind if we examine the research work of John Gregor Mendel, we
find that success of this scientist was because of the material pea (Pisum
sativum). Because the seven characters (genes) are located separately
on separate chromosomes.
On chromosomes there is large number of genes. For example in
Drosophila melanogaster on 8th chromosome there are more than
100 genes, in human being on 46th chromosome according to Stern (
1949) there are about one lakh genes. This fact reveals that in every
pair of homologous chromatid there are many genes. It also happens
that two or more genes responSible for a character are located on
chromatid of pair. Such genes are linked in such a way that all such
genes together reach to gamete through meiosis. Such relationship
between genes is known as linkage.In homologous chromatid
presence of total number of linked genes is called linkage group.
Discovery of linkage was done in 1906 by Bateson and Punnet, but
they could not explain it. In 1910 Morgan did similar experiment in
Drosophila melanogaster. Results of Morgan were similar to results
of Bateson and Punnet. Morgan explained the reason as linkage. He
explained that alleles (genes) from homozygous parent reach together
in same gamete, but if same alleles (genes) come from heterozygous
parent then they reach separately in different gametes. The
87
phenomenon of existing of genes together in a gamete has been named

as coupling by Morgan. The phenomenon of dispersing genes
independently in different gametes has been named as repulsion.
Morgan expressed that coupling and repulsion are two phases of
linkage process. Explaining the reason of coupling Morgan told that
the reason for linked genes to be together is they are annexed on one
chromatid only. Explaining repulsion Morgan told that because the
alleles (genes) exist on separate chromosomes therefore they go to
different gametes. Morgan also told that quantity of linkage is
inversely proportionate to distance of linked gene. After the discovery
of Morgan scientists could feel that genes are arranged in line on
chromosomes. Based on this theory of Morgan it could be possible to
prepare linkage map (genetic map), by its help location of genes on
chromosomes can be understood.
Explanation of linkage through experiment:
Phenomenon of linkage can be well understood by breeding
experiment on maize (Zea mays) and Drosophila flies (Drosophila
melanogaster) because in their offsprings characters can be understood
well. Moreover they produce offsprings in large numbers. Therefore
it is easy to make ratio. Details of experiments are given below:
Experiment with maize:
Two contrasting characters of maize are coloured & full seeds and
colorless & shrunken seeds. These characters can be expressed like
this:
Colored = gene C Full seed = gene S Colorless = c
Shrunken seed = s
Therefore genotype of coloured & full seed is CCSS and genotype of
colourless & shrunken seed is ccss. These two when produce genotype
produce the gametes CS and cs. On crossing these the hybrids
obtained has the genotype Ccl Ss (linked genes are written above
and below the line). The F1 will produce coloured and full seeds. As
per the Gregor Mendels law of independent assortment the F1 will
produce 4 types of gametes viz. CS, Cs, cS, cs in equal numbers. But
when this hybrid was test crossed (crossed with recessive parent)
then in F2 the ratio of 1:1:1:1 could not be obtained, which should
have happened. The obtained ratio was different i.e. 29 coloured &
full: 1 coloured & shrunken: 1 colourless & full : 29 colourless &
shrunken ( 29: 1: 1: 29)
88
Linkage
In another experiment when coloured & shrunken seeded (Cc ss)

maize plant was crossed with colourless and full seeded plant. Then
in F1 the offsprings were of four types in the ratio 33:1 :1:33 ( 33
coloured & shrunken = Cc ss : 1 coloured & full seed = CcSs : 1
colourless & shrunken = ccss : 33 colourless & full seed = ccSs ).
Both above mentioned experiments reveal that colored & full and
colourless & shrunken in the experiment A and coloured & shrunken
and colourless & full in experiment B are more popular. This happens
because of following causes.
(1) Two pairs pf genes i.e. Cc and Ss could not assort independenly.
(2) During meiosis these pairs of genes are arranged in parental
combination and recombination.
(3) In homozygous (CCSS) as well as in heterozygous (CS /cs)
.,1 ...
.' • ::D
.. '
~
I Form
l. .JUndl0n
=,.,=
,. :.=:,=x.;.~ 11 =
Branch
1! Migrate
=':::x : ::D
'0. SI
e
~~;:::~~i~
.1 ":1*
Resolve ~
,.• <:
•
<
•
=-
,
"al-
+
, "aI..:-
::D
,.,. .t::
Llgate
~
• ; .::D
<
•• ,,_"aI, 'ta •
.:5
S.
"al.
Patch
1\. CD.
Sp1i(;e
, 11
Fig. 19: Recombination of genes.
89
parental combinations are much more i.e. 90 to 95 per cent. The

reason may be that parent genes are closely linked.
(4) In F2 the new characters such as coloured & shrunken (Cs I cs)
and colourless & full ( cSI cs) are visible due to recombination
in experiment A. Similarly coloured & full (cSI cs) and colourle-ss
& shrunken ( csl cs) are visible due to recombination in
experiment B.
Mechanism of recombination is shown in ( Fig-19).
Complete linkage:
If two or more characters are inherited together and continue to be
inherited together in next 2 or more generations then those characters
express complete linkage.
Complete linkage is very rarely found. Example is male Drosophila
melanogaster (female do not show complete linkage). Another example
is female silk worm. An other example of complete linkage is Sorghum
vulgare. The phenomenon of complete linkage is being explained by
following examples.
Example to explain complete linkage:
Complete linkage can be well understood by a breeding experiment
in Drosophila melanogaster. If normal grey bodied & long winged female
(genotype GGLL) is hybridized with black bodied & vstigeal winged
male (genotype ggll). In Fl all the offsprings will be grey bodied and
long winged (genotype GgLl). If this Fl (GgLl) is crossed with black
bodied & vestigial wing homozygous recessive (genotype ggll). Then
in F2 generation grey bodied & long long winged and black bodied
& vestigial wing will be produced in equal numbers (1:1 ratio).
Explanation of the experimental results:

Results of the above experiment can be explained with cytological
background. It is clear that male of Fl produces only two types of
sperms - GL and gl in equal numbers. But the homozygous double
recessive female produce only one type of gamete - gI.
Therefore production of two types of gametes (GL and gl) by
heterozygous male of Fl, are infact contrasting characters and are
closely linked with each other. The reason is that G and L are situated
on same chromatid. Similarly g and I are situated on other same
chromatid.
90
Linkage
Hence two types of sperms .. GL and gl and one type of egg - gl

when fertilize produces only two types of offsprings which resemble
only to their parents. Because these characters have complete linkage,
therefore new recombinations were not be produced.
Instead of homozygous dominant parent ( GL/GL) when
homozygous recessive parent black bodied & long winged (gl/ gl)
was chosen for crossing with grey bodied & vestigial winged ( Glf
gl) flies, then in the F1 these two types of flies are produced in equal
numbers ( 1:1).
Incomplete linkage:
If two or more characters come together in offspring but due to
recombination produce new character in such case those characters
are said to express incomplete dominance. As compared to complete
linkage the incomplete linkage is more common in plants. From the
following example incomplete linkage can be understood well.
Examples to explain incomplete linkage:
In the above mentioned experiment on Drosophila melanogaster in
F1 if grey bodied long winged heterozygous female fly ( genotype
GL/ gl) is crossed with double recessive ( glf gl) black bodied wing
male then the F2 will produce four types of offsprings . Out of this,
two parent type combinations will be more in number. Other two
types of combinations will be of new types. The four types of
offsprings will be 41 % grey bodied & long winged, 9% grey bodied
& vestigeal winged, 41 % black bodied & vestigeal winged and 9%
black bodied & long winged.
Instead of homozygous parent if mutant parents - black bodied &
long winged (genotype gL/ gL) are crossed with grey bodied &
vestigial wing ( genotype Glf GI ), then the F1 generation will be
grey bodied & long winged with a genotype Glf gL.
If F1 female with genotype Glf gl crossed with male of the genotype
gl/ gl then in F2 three types of offsprings are found i.e. 41 % grey
bodied & vestigial winged 9 % black bodied & vestigial winged 41
% black bodied & long winged.
Causes of complete and incomplete linkage:
(1) One main reason for complete linkage is that in F1 offsprings of
normal heterozygous or mutant heterozygous parents, the
alignment of gene without any change in original condition
91
reaches to gametes. The result is that chromatid of homozygous

parents having the genes GI & gL and the chromatid of mutant
parent having the genes GI and gL without any change goes to
their gametes during Fl. Therefore heterozygous Fl male
produces two types of gametes. Due to which in F2 two kinds of
offsprings are produced in equal number.
(2) Second reason according to Morgan is the exchange of one
segment of a chromatid with other segment of chromatid. Such
exchange takes place in first prophase of meiosis. Generally it
happens in non homologous chromatids. Because in this activity
there is exchange of genes from one chromatid to another,
therefore this activity is called crossing over. Therefore one of
the main reason for incomplete linkage therefore is crossing over.
Strength of linkage:
Strength of gene linkage depends on distance between genes on
chromatid. In other words what is the frequency of crossing over
there. For example if two genes Aa and Bb are a distance of 10 units
and other gene pair Cc and Dd are at a distance of 4 units. In that
case the linkage in Cc and Dd will be stronger than linkage in Aa and
Bb. In other words it can be said that the stronger linkage leads to
lesser crossing over. Similarly weaker linkage leads to possibility of
more crossing over. Therefore it can be said that strength of linkage
depends on possibilities of crossing over.
Factors affecting strength of linkage:
As described above linkage strength depends upon distance between
genes located on chromosomes. Beside it there are following
physiological and environmental factors affecting strength of linkage.
(1) Age: Strength of linkage increases with increase of age.
(2) Temperature: Increase in temperature results reduction in
strength of linkage.
(3) Ionizing radiation: Increase in dosage of x rays, gamma rays,
etc. reduces of strength of linkage.
(4) Chemicals in food: Presence of chemicals in food for very long
time may weaken the strength of linkage.
Measurement of linkage strength :
To measure the strength of linkage, the organisms having two or
more linked genes are hybridized. In the Fl heterozygotes are
92
Linkage
obtained. This heterozygote is crossed with homozygous double

recessive parent. F2 has offsprings with parental combinations. These
are counted separately and the percentage is calculated. Percentage
of new recombination is also found out- this suggests the strength of
linked genes. More the percentage of new recombination lesser the
strength of linkage.
Importance of linkage:
(1) The discovery of linkage by T.H. Morgan has important place in
genetics.
(2) Based on linkage some new genetical concepts were generated
and developed.
(3) For improvement of crops, vegetable, fruits etc. to plan out the
programme, understanding of linkage is very helpful.
93
Chapter 11
Crossing Over
In earlier chapter it has been explained that during first prophase of
meiosis two chromatids of homologous chromosomes pair with each
other at several places. It results in exchange of segments. This type
of pairing of chromatids is called crossing over. During pairing
chromatids gets broken where linkage is weak. Broken parts of a
chromatid join with broken part of other chromatid. Therefore
crossing over can be defined as: at pachytene stage of meiosis, the
inner chromatids of homologous chromosomes pair with each other
and get broken at pairing place. Recombination of broken places is
called crossing over.
Somatic & germinal crossing over:
Depending on type of cell crossing over is of two types. If crossing
over occurs in somatic body cells then it is known as somatic crossing
over. If crossing over occurs during gamete formation, thenit is called
germinal crossing over. It is very important in heredity. Therefore it
is discussed in this chapter. Details of somatic crossing over are
described below.
Somatic crossing over:
In development of embryo during mitosis pairing between chromatid
leading to crossing over is termed somatic crossing over. It occurs
very rarely. In such tissues there is a mosaic pattern of cross over and
non crossover. Somatic crossover can not be inherited so it will not
be discussed here.
Germinal crossing over:
At the time of gametogenesis during meiosis germinal crossing over
takes place. It happen at pachytene stage. Germinal crossing over is
common in organisms. It occurs more in heterozygous chromatids as
compared to homozygous chromatids.
94
Crossing Over
Mechanism of crossing over:

You have already studied that germinal crossing over happens in
early prophase of meiosis. At this time two homologous chromosomes
come near each other and do pairing (or synapsis). Afterwards both
the paired chromosomes replicate to form two chromatids separately.
Hence the chromosome pair now has four chromatids. At this stage
the inner two chromatids twist on each other. The place where the
two chromatids lie on each other is called chiasmata. Mostly on one
place the pairing chromatics breaks. Afterwards one broken part joins
with other broken part. This results that new segments appear in the
chromatids. Afterwards these chromatids move away from each other.
In last all the four chromatids move to four gametes. Out of 4 gametes,
in two gametes there are normal (non cross over) chromosomes and
in rest two gametes there are recombined (crossover) chromosomes.
Therefore these gametes are also known as non crossover gametes
and cross over gametes ( Fig-20).
Prophaael
ofmefosJa
......... 1
Fig. 20: Mechanism of crossing over.
95
Cytological explanation of crossing over:

Cytological explanation first of all was given by C. Stem on Drosophila
melanogaster and B. M. Clintoch on Zea mays (maize)
C. Stem observed red and round eye in ordinary Drosophila
melanogaster. However in case of mutant eye of Drosophila there was
different colour and shape. Deep red eye colour is controlled by
recessive mutant and bar eyed is controlled by dominant mutant gene.
Both these mutant genes are present on x chromosome. This X
chromosome in Drosophila melanogaster is rod shaped. In female fly
there is a pair of X chromosome. In his experiment Stem observed
.;..;.~
--
¥frogmon.. -
x
... +
v_tl + f
-
\"1_.
-/
.. x
1..-
1
~-
;&
..-
~ -
\
ciF
• ;; i
L~ ""£1-
€9 (,;,.,:.
-=£1....
...
-
x_
L~~ _NI ~-=-

<1£
(r i
€~~ -=, ., -will.".
F,_
Fig. 21: Cytological proof of crossing over.
96
Crossing Over
such a fly in which one x chromosome was divided in two parts.

Broken parts had mutated genes Carnation and Bar. These were
located on broken points (Fig. 29). On the end of another X
chromosome there is attached a fragment of Y chromosome. These
abnormalities happened because of sudden chromosome breakage. Y
fragment attached X chromosome has normal allele of Carnation and
Bar genes. Crossing over happened in both this x chromosomes during
meiosis. With the result Y fragment of X chromosome gets transferred
to broken x chromosome. Hence crossover chromosome and non
crossover chromosomes differ in their morphology which can be
observed under microscope. An example is described in Fig-21.
Theories of crossing over:
Though crossing over is a precise process but it is not clear that what
the reasons for crossing over are. To explain this phenomenon several
theories were put forth, but following theories are most popular:
1) Contact first theory:
It was proposed by Serebrovsky According to this theory the inner
chromatids touch each other at one or many places and do the process
of crossover. At places of touch the chromatids have breakages.
Broken segments of the chromatids join with each other and form
new combinations (Fig- 22)
2) Breakage first theory:
This theory of crossingover was given by Muller. According to this
theory chromatids break without doing crossing over. Afterwards
broken segments of the chromatids join with each other to yield new
combinations. Nowadays this theory is accepted by many scientists
(Fig-23).
3) Strain theory:
Strain theory of crossing over was given by Darlington. This theory
explains that the reason for breakage is the tension due to pairing.
Both the chromatids of homologous pair have spiral coiling on each
other. It results tension. Due to which at point of contact chromatid
breaks. Afterwards these broken ends recombine leading to
recombination of genes.
97
Fig. 22: Contact Fig. 23: Breakage

theory of theory of Crossing
Crossing over. over.
Factors affecting crossover:
Following factors affect crossing over:
1) Internal factors:
(A) Sexual difference: Amount of crossing over differs in male and
female sexes. For example males of Drosophila melanogaster do not have
crossing over because it has complete linkage. In females of silkworm
moth (Bombyx mon) there is no crossing over. It has been observed
that if in organisms both the sexes exhibit crossing over then it happens
in equal numbers e.g. peas (Pisium sativum). In some females crossing
over happens more than males example mouse. Sometimes reverse is
true i.e. crossing over happens morein males as compaired to females
example pigeon. Haldaneons (1921) reported that wherever crossing
over is different in different sexes then heterozygous sex express less
crossing over.
98
Crossing Over
(B) Modification in chromosomes: Change in structure of

chromosome results in reduction of crossing over. Change in
chromosome happen due to inversion, translocation, duplication,
deletion etc.
(C) Mutation: Occurrence of mutation results in reduction of crossing
over.
(D) Age: With increase in age there is reduction in crossing over.
(2) External factors:
(A) Temperature: Reduction in temperature leads to increase in
crossing over. Medium temperature reduces this increase in
crossing over. At high temperature crossing over does not occur
or is negligible.
(B) Ionizing radiation and chemicals: Ionizing radiation such as
treatment with X ray, gamma rays etc. or treatment with
chemicals increases frequency of crossing over.
Importance of crossing over:
Crossing over happens in bacteria, yeast, fungi, higher plants and
animals. Crossing over has following importance:
(i) Because of exchange of segments crossing over results
recombination of characters.
(ii) Studies of crossing over reveals that genes are arranged in length
of chromosomes.
(iii) Chromosome maps are made based on frequency of crossing
over.
(iv) Crossing over helps to understand nature and mechanism of
genes ..
(v) Crossing over has lot of importance in plant breeding.
Importance of crossing over in breeding:
(i) Crossing over is very important in breeding of plants and animals.
Breeder gets recombination because of crossing over in
hybridization.
(ii) It is necessary to remove undesirable or uneconomical characters
which may occur in .hybridization, but due to occurrence of
linkage the removal of undesirable characters become not
possible because those are linked with desirable characters.
Therefore because of this close linkage recombination can not
occur. Therefore to over come this problem treatment with
ionizing radiation (X- ray, gamma ray etc.), heat, chemicals etc.
are tried.
99
Chapter 12
Chromosome Map
Before the knowledge of linkage and crossing over it was only known
that chromosome contains genes. But nothing could be known about
location of genes. Morgan, Bridge and Sturtevant based on their
experiments have given following conclusions:
(1) In chromosomes genes are located in linear fashion.
(2) Linked genes do not show recombination in normal
circumstances.
(3) Frequency of recombination and strength of linkage depends on
distance between genes.
(4) Every gene is located on locus (plural loci) situated on
chromosomes.
(5) In linked gene recombination occurs to the extent of 1 % to 50 %.
Based on the above mentioned facts geneticists have tried to draw
chromosome map. In this map based on calculations of crossing over
percentage, distance between genes are located on a straight line on
a paper. Therefore chromosome map can be defined as chromosome
map is a line on which location of gene is decided based on crossover
percentage.
Example to draw chromosome map:
In a trihybrid F1 the heterozygous AaBbCc was back crossed to the
homozygote aabbcc parent. Resulting progenies were the followings:
Genotype Number
AaBbCc 79
AabbCc 18
AaBbcc 82
Aabbcc 21
aaBbCc 19
aaBbcc 23
aabbCc 80
aabbcc 78
400
100
Chromosome Map
Prepare a chromosome map showing the order of genes and their

distance.
Method to prepare chromosome map:
Parental types are selected from all the progenies mentioned in the
question. For selection the easy way is the progenies havit:tg maximum
number are the parental types. Therefore in this example the parentals
are mentioned below:
Genotype Number
ABc 82
abC 80
Afterwards recombination percentage obtained from two genes is
found out. To find it the test cross used is known as two point test.
Calculation of recombination percentage from A and B genes
Because both genes come from same chromosome, therefore they
express coupling phase. Given below the side two combinations are
parental types and the central two combinations are recombination
types:
AB Ab aB ab
79 21 19 80
82 18 23 78
161 39 42 158
Two
recombination
types
Recombination = 39+42 = 81
Total population = 400
Recombination percentage = 100x81
_ _ _ =20.25%
400
Calculation of recombination percentage from Band C genes:
Because Band C genes comes from different genes, therefore they
express repulsion phase. Therefore in the combinations given below
101
the side two are recombinations and the central two are parental
type combination.
BC Bc bC be
79 78
19 21
98 99
Parental recombination
Recombination = 98+99 = 197
Recombination percentage = 197 x 100
= 49.25 %
400
Calculation of recombination percentage from A and C genes
The genes A and C are from different chromosomes, therefore they
express repulsion phase. In the following combination the two types
of corners are parental types and the central two types are
recombination types.
AC Ac aC ac
79 78
18 23
97 101
Parental types
Recombinations = 97 + 101 = 198
Recombination percentage = 198 x 100
=49.5 %
400
AB = 20.25 % recombination
BC = 49.25 % recombination
AC = 49.5 % recombination
102
Chromosome Map
49.5 %
A 20.25 % B 49.25 % C
49.5 %
A 20.25 % b 49.25 % c
From the above mentioned two point tests, sequence of genes on
chromosomes is known, but distance is not known between genes.
The reason is that between A and B & B and C there exist double
crossing over value. On subtracting double crossover value from the
two [email protected] the distance between A and B & B and C can be known.
To find out double crossover value we have to follow three point
test described below:
Non crossover ABc = 82
Or parental type abc = 80
162 divided by 4 = 40.5 %

Cross over between A and B
aBc = 23
AbC= 18
41 divided by 4 = 10.25 %
Cross over between B and C
ABC = 79
Abc = 78
157 divided by 4 = 39.25 %

Crossing over between A and C or double cross over value ( Fig-24)
aBc = 19
Abc = 21
40 x 10/400 = 10 %
103
~c _ _ _ _ iO"-'-C
A
•I C A
• C A, fI ,
C
:s I I 4IIIC
•I
I
}=-
- -
b c (-
-: I I
A,
•
, C
,
-Wo)
)<
"
I
- •
4IIIC
-= •, c,
}a€:l
~ _ _ _ _ ."-A-C
x::
fI A
I i " A C
-= /J, A
!
C
I
}::=-.
)<
1
, - --=-=
-=
b,
/J
I
•I
•I
A,
1
,
C
•, h?,:,
•
~
5C
1 ~
,
_ _ ..... _IoA-C-'
-=: 1t i :t }~_
,- - -=-=-= f
1 1
f
)< , !
i t r:=~
.. " • c
Crossing over After crossing over

Fig. 24: Crossing over between A and C or double cross over.
On subtracting double cross over percentage between A and C Le,

10% from the cross over percentage calculated by two point test
distance between A & B and B & C can be known,
Note: Percentage value has been calculated from total population (i.e,
400),
According to three point test:
AB = 10,25
BC = 39,25
AC = 39.25 + 10.25 = 49.5%
Now after deciding scale, chromosome map can be made as decribed
below:
Scale: 10% = 1 centimeter
104
Chromosome Map
Fig. 25: Chromosome map of four chromosomes of Drosophila

melanogaster.
105
10.25% 39.25% 49.5%

A B C
10.25% 39.25% 49.5%
a b c
Therefore it is the desired chromosome map which shows sequence
of genes and distance between the genes.
Geneticists have made chromosome map of several organisms. For
example chromosome map of Drosophila melanogaster in Fig-25.
Importance of chromosome map:
Chromosome map is important because of following reasons:
(1) Chromosome map gives details about exact location of genes,
arrangement of genes and combination of genes.
(2) By study of chromosome map it is possible to guess results of
hybridization.
(3) Chromosome map can be helpful as probability table.
(4) Chromosome map helps in breeding work because it gives details
of genetic combination.
106
Chapter 13
Chromosomal Aberrations
Structural changes in chromosomes (Chromosomal aberrations):

Any deviation in normal chromosomes architecture or ploidy level is
known as chromosomal aberration. Chromosomal aberrations are of
two types - (1) structural aberration, (2) numerical aberration. Both
aberrations can occur during any stage of cell division due to certain
circumstances.
According to Morgan's theory of chromosome, in every organism
there are fixed number and structure of chromosomes. Genes are
arranged there in linear order. These chromosomes are inherited to
their offspring in definite number and structure. But because of certain
internal and external factors chromosomes gets some changes. It
results in changes of characters. These changes are important in
development of new types. It may give evolutionary benefit
occasionally of new genotypes.
How chromosomal aberrations or structural changes are produced:
A small broken part of chromosome is called fragment. A fragment
without centromere is unable to move to poles. Such fragment without
centromere is called acentric fragment. It is well known that
centromere is an important part of chromosome because it helps in
movement of chromosomes to poles during cell division. If due to
certain circumstances chromosomes gets broken and are joined again.
It may result following modifications in chromosomes:
(1) Any broken segment can join with a centric segment.
(2) Any acentric segment can join with centric segment.
(3) Two acentric segments can join with each other.
Out of 3 situations described above only the second one survive.
Rest both situations can join with each other and perish.
Types of chromosome aberrations:
Chromosome aberrations are of following types:
107
(1). Deletion or deficiency

(2) Duplication.
(3) Simple translocation.
(4) Reciprocal translocation.
(5) Inversion.
These are described below.
(1) Deletion or deficiency: Either of two ends of chromosome due
to certain cause separate out and forms two parts. One part is
monocentric i.e. have one centromere and other part without any
centromere or in other words acentric. This situation is called delition.
It is a simple chromosomal aberration. In this only one chromosome
is affected. Such change is less hai-mful for the organism. Characters
of offspring are less affected by deletion. Deletion plays a great role
in evolution. It is of following two types.
(a) Terminal deletion: If any terminal segment gets broken and
destroyed then it is called terminal deletion. If a chromosome
consists of abcdefgh and the ab gets separated then only cdefgh
remain left over. In the ab portion there is no centromere. So it
can not move to either pole. Hence it gets ultimately lost. (Fig-
26)
(b) Intercallary deletion:
From in between chromosome, segment may break off. Afterwards
the broken chromosome again joins. This situation is known as
intercalary deletion. As for example from abcdefgh chromosome cd
segment breaks away. So the ends b and e join together. The segment
cd does not have centromere, so it can not move to any pole and
therefore get destroyed. (Fig-26)
Normal chromoS()Dle
AB CDEFGH
/\ 0--
_A_BIIIi~_C_D_E~Fd "'A B\C ~\E F ~

J E~
----0--
C D F G H
Terminal deletion
----c:.-
A B F G H
Interstitial deletion
Fig. 26: Different types of deletions.
108
Chromosomal Aberrations
(2) Duplication: In chromosome if a segment gets replicated then

this phenomenon is called duplication. Number of genes increases
(gene amplification) because of duplication. This situation sometimes
results lethal effect on the offsprings. Duplication may be of two types
i.e. terminal or intercalary.
(3) • Simple translocation: In simple translocation atleast two
chromosomes are affectea. From one chromosome a segment breaks
off and joins with other chromosome. This translocation affects the
inheritable characters. In evolution it has great importance. Simple
traslocation can be understood well with the help of Fig-27. From the
wxyz chromosome yz segment breaks off and joins with the abcd
chromosome giving it a new genotype abcdyz.
(4) Reciprocal translocation: It differs from simple translocation. In
case of reciprocal translocation some portion (segment) of both the
chromosomes are broken off and then joined to each other
chromosome. These change the structure of both the chromosomes
and results in change of characters. It is important in evolution. It can
be well understood by Fig-27. From abcdef chromosome ab segment
is broken. Similarly from vwxyz chromosome vw segment gets
broken. Afterwards these broken segments get joined with each other
chromosome making the new genotypes as abxyz example
Philadelphia chromoso~e.
- . _I::. +
Fig. 27: Different types of translocations.

Differences between reciprocal translocation and crossing over
There are following three differences:
(1) Main difference between reciprocal traslocation and crossing
over is that in crossing over transfer of chromosome segments
occurs only in between inner chromatids of a chromosome where
as reciprocal traslocation occur in whole chromosome. It is not
essential that the chromosomes are homologous.
109
(2) Another difference is that in crossing over formation of chiasma

occurs where as in reciprocal translocation chiasma do not occur.
(3) Crossing over is an essential process but reciprocal translocation
is an accidental and unplanned process.
(5) Inversion: In chromosomal inversion only one chromosome is
affected. In the chromosome, sequence of genes is changed. Its
affect is not lethal. Hence inversion has great scope in evolution.
There are following two differences in inversion and translocation:
(i) In case of translocation structure of chromosomes gets changed,
but in case of inversion structure does not change at all. Only
there is a change in sequence of genes.
(ii) In translocation at least two chromosomes are involved but in
case of inversion only one chromosome is involved.
Types of inversions:
Inversion is of following two types:
(i) Terminal inversion: Sequence of genes is changed at end of
chromosome. The sequence of genes get changed to abedc from
abcde.
f\ .6 C _n E
• "I Pa1'i1oontrlc
A BeD E
c." r~," -"....:~;:: :,:=::.~::::_,~-.:_:~ ~" Palacenlric - ~.~[)""'jIII01IlIiI!1!III!I".
Ii, ll'V~"""1 MttIro4'~
a: A 0 C B i invefllion heterozygote
"Po",'1I
J Crt.I88CM'.1f
c
-."_,BAD.
Ittktq)
Palnng
c ~"h~~
D Two cl'03S0vers In loop
~ Stlgl'f)gatfOl'l
•
I A B G " f
l Segregalion
./10'
e, 0
t~ ~\IIN)
B "
At~rlr:
fragment
A ,
Normal product
A BeD E
Double crossover product c.,_,,:-:::::::-=_-_-::::.:'
OOUb!ecrossoverprodl,lCl
A 0
_~:::::-
CBE
__
ADC B E
Fig. 28: Types of inversion and inversion loop.
110
Chro11loso11lal Aberrations
(ii) Intercallary inversion: In this case instead of at end, the sequence

of genes changes in between the chromosome. The sequence of
genes instead of abcde it changes to acbde. If inversion occurs
in one chromosome out of two homologous chromosomes then
at time of pairing it forms inverted chromosome loop as shown
in Fig-28.
Evolutionary significance of chromosomal aberration
(1) Due to chromosomal aberration there is a change in genotype.
Therefore phenotype gets changed. .
(2) If any organism has homologous deletion, then it has lethal or
fatal effect. But due to duplication this harmful effect is
neutralised.
(3) Usually duplication results increase in chromosome number
which has evolutionary significance.
(4) Inversion and translocation play a great role in evolution because
it results semi - male sterility or it changes the character.
111
Chapter 14
Cytoplasmic Inheritance
Nowadays this type of inheritance, governed by non nuclear factors
are refered as extrachromosomal inheritance, somatal inheritance,
extranuclear inheritance, maternal inheritance, uniparental mode of
transmission or transovarian transmission.
By definition, it is the inheritance controlled by the extra chromosomal
i.e. cytoplasmic factors that is transmitted to the succeeding generation
through the egg of female organism.
It does not follow Mendelian inheritance but have following
characters.
(a) It does not follow Mendelian segregation in crosses and reciprocal
crosses.
(b) A particular set of characters are controlled by a set of cytoplasmic
factors (genes) producing dissimilar hybrid.
(c) Follows the maternal1ine i.e. uniparellel mode of transmission
These cytoplasmic factors are contained by:
(i) mitochondria (mt DNA)
(ii) chloroplast ( cp DNA)
(iii) symbionts ( bacteria and virus)
It has been explained earlier that inheritance happens due to genes.
Genes are located on chromosome located in nucleus. Now a question
may be raised that is there any method of inheritance that does not
involve genes? The answer is yes. In modem times it has been proved
that certain gene like particles present in cytoplasm are responsible
for inheritance of some characters. These particles are called plasma
gene. Therefore the mechanism where plasma genes effect inheritance
is called cytoplasmic inheritance. Plasma genes and nuclear genes
express relationship.
Maternal effect:
It is well known fact that the amount of cytoplasm in female gamete
is much more than male gamete. Therefore by cytoplasm the female
112
Cytoplasmic Inheritance
gamete inherit more characters. The female gamete is responsible for

transfer of cytoplasm to zygote. Therefore it can be said that female
gamete express maternal effect for example ephertia moth.
Extranuclear inheritance:
There is enough experimental evidence to show that for inheritance
not only nuclear genes but extranuclear genes are also responsible.
This phenomenon is known as extranuclear inheritance or cytoplasmiC
inheritance. The examples are mitochondria, chloroplast inheritance.
These items exist outside nucleus and are inherited independently
from generation to generation and also called organelle inheritance.
Examples of cytoplasmic inheritance:
(1) Cytoplasmic inheritance in plants:
(a) Plastid inheritance: Good example of this phenomenon is
variegated Mirabilis jalapa. In this plant some branches have
variegated leaves. The variegation of leaves is controlled by some
plasma genes present in cytoplasm. It is not concerned with
chromosome. Similar inheritance of plastid is reported in beans,
maize etc.
(b) Pollen sterility in maize: Working on maize Rhoades in 1933
reported a mutant race. This mutant race produced sterile pollen
grains. When this mutant race was hybridized with normal fertile
race then it could be known that the genes responsible for pollen
sterility were inherited by cytoplasm present in egg cell.
(c) Inheritance in rusts: Rust is caused by parasitic fungus which
causes disease in wheat and other crops. Inheritance of the fungus
takes place by chromosome. Sometimes its inheritance is
controlled by plasma genes.
(d) Inheritance in Epilobium: On hybridization of Epilobium luteum
as female with Epilobium hirsutum as male. The F1 offsprings are
fertile. But when the position of female and male was reversed
(reciprocal cross), then the F1 produced sterile pollen. The
experiment reveals that the pollen sterility gets inherited by
cytoplasm of Epilobium hirsutum.
(e) Cytoplasm and hybrid vigour: Dhavan, Bhatt and Jagi (1965)
have reported that in maize the cytoplasm controls hybrid vigour.
113
(2) Cytoplasmic inheritance in animals:

(a) Cytoplasmic inheritance (transmission) in Drosophila: Heritier
and Teissier (1951) have reported that ordinary Drosophila
(Drosophila melanogaster ) is resistant to carbon dioxide, but one
particular race is very sensitive to carbon dioxide. The reason is
presence of plasma genes. Carbon dioxide sensitive Drosophila was
crossed with cabon dioxide resistant Dros'ophila. The female
offsprings were hybridized with ordinary Drosophila. On repetition
of this hyb~idization the offsprings were sensitive to carbon dioxide.
This character is inherited by the cytoplasm of carbon dioxide
sensitive fly.
(b) Cytoplasmic inheritance in Paramecium: Sonneborn (1949)
reported a killer race of Paramecium aurelia.1t produces toxic substances
so kills other race of Paramecium aurelia . This killing character of
Paramecium aurelia is controlled by Kappa Particles gene like material
present in cytoplasm. Ordinary race of Paramecium aurelia do not have
Kappa particles.
Genetic examinations have revealed that kappa particles are related
to gene present in chromosome. In absence of this gene Kappa particles
do not grow.
Role of cytoplasmic inheritance in evolution:
Darlington and Write have expressed that cytoplasmic inheritance
has secondary role in evolution. However, Lamprecht (1944) has
reported lot of importance of cytoplasmic inheritance because it may
develop lethality and sterility. Moreover plasma genes, like nuclear
genes, express mutation. Therefore plasmagenes are as important as
nuclear genes in evolution.
114
Chapter 15
Sex Determination
Male and female two sexes are most popular. These can be identified
by morphological, anatomical and physiological characters. These
characters can be primary and secondary as described below.
(1) Primary sex structures: Primary sex structures are directly related
to gonads, ovaries and testes. In higher female animals it is in
form of vagina, uterus and oviduct. In males it is in form of
penis, vas deferens etc.
(2) Secondary sex structure: Under secondary sex structures we
consider accessory sex organs for example human females have
more growth of breast. In male breast growth is less. In addition
to it males have beards and mustache where as female females
do not have these.
In humans and animals male and female sexes are present separately.
The same phenomenon is also observed in some plants. But many
plants are hermaphrodite.
Sex determination:
Sex determination differs from sex differentiation. According to
Darlington and Mathur (1949) sex determination is a process by which
male and female gametes give rise to male and female organisms. Sex
differentiation is the anatomical, morphological and physiological
changes during development of organism.
Before the knowledge of inheritance several biologists have attempted
to know sex determination. Before 1900 in this connection some
hypothesis were also put forth. But attention was more to external
environment than inheritance by reproductive cells. Some scientists
thought that sex of a child depended on relative vigour of their father
and mother. If father is more vigorous then male child and if mother
is more vigorous then female child is borned. Some scientists
expressed that sex of a child was dependent on relative age of father
and mother. Some scientists said that sex of a child depends on
nutrition given to the mother. Some scientists opined that time of
115
copulation of parent had effect on sex determination of child. Some

scientists have expressed opinion that sperm and egg producing organs
(testes and ovary) have effect of sex determination of child. On
discussion none of the ideas expressed above proved successful.
Time of sex determination:
As mentioned below sex determination can be possible in three times:
(1) Progamic: Before fertilization of egg cell.
(2) Syngamic: At fertilization.
(3) Epigamic: After zygote formation.
Theories of sex determination:
For sex determination two types of theories have been proposed:
(1) Genetic theories: Bateson, Emersion, Correns and other scientists
have accepted the genetic theory.
(2) Physiological theories: Riddle and Goldschmat are the main
scientist who accepted the physiological theory.
The above mentioned theories are enumerated below:
(1) Genetic theories:
(a) Alternate dominance theory.
(b) Heterogamosis theory or chromosome theory.
(c) Genic balance theory.
(2) Physiological theories:
(a) Metabolic differentiation theory.
(b) Quantitative theory.
These theories are discussed below in some detail:
(1) Genetic theories:
(1 a) Alternate dominance theory:
Alternate dominance theory was suggested by Castle. According to
it in fertilization there is union of male and female gametes. From
one type of gamete fertilization will not be possible. Hence for sex
every gamete acts heterozygous. Castle also stated that dominance is
also expressed in sex characters and for sex determination there are
some determiners During fertilization if male gamete determiner is
dominant then offsprings are male. In case if the female gamete
116
Sex Determinatio1l
determiner is dominant then the offsprings will be of female sex. In

modem time this idea is not accepted.
(1 b) Heterogamosis or chromosome theory:
This theory was presented by Correns. He pointed out that in
fertilization out of the gametes uniting is heterozygous for one sex
and homozygous for the other sex. Correns has proved this theory
by cytological and genetical experiments.
Sex determination by chromosomal mechanism:
Sex determination by chromosomal mechanism was proposed first of
all by Mc Clung (1902). Working on cytology of grass hopper Mc
Clung has seen that in body cells of female there are all pairs of
chromosomes but in male there is only one chromosome alone i.e.
without any pair. In other words number of chromosomes is even in
female but uneven in male. On this observation Mc Clung suggested
that because in males number of chromosomes is odd, therefore during
meiosis two types of gametes should be formed. In half number of
gametes chromosome number should be odd and in other the
chromosome number should be even. Mc Clung stated that in case of
female grasshopper all the gametes should have even number of
chromosomes. Moreover after fertilization with one type of male
gametes the offsprings should have one type of sex. Similarly after
fertilization with other type of male gamete the offspring should have
other sex. The ratio between sexes should be 1:1. Therefore Mc Clung
told that odd chromosome number is the sex determiner.
In 1905 E. V. Wilson and N.M. Stevens proposed some proofs to
confirm chromosome theory of sex determination. He designated
heterochromosome as X chromosome. He expressed that in male only
one X chromosome is present but in female two X chromosome are
present. Naming the absent X chromosome as 0 Wilson expressed
the XO system. (Fig-29)
After few years Wilson again did experiments on sex determination
on other insects. He told that though male and female have same
number of chromosomes. However in female there are two x
chromosomes but in male there is only one X chromosome. The other
pair is very small and is called Y chromosome. Therefore on this basis
the female has xx chromosome and the male has XY chromosome.
So from both angles the male has heterogametic or heterozygous
gametes. Sex determination of any organism is done at fertilization.
117
If X chromosome of male fertilize the egg cell then it produces female

sex. But if Y chromosome of male fertilize the egg cell then male sex
is produced (Fig-29)
ne xx-xv and zw:zz. type ~ oh... ;., Haploid lIIIIle, diplool female oM xx-xo type-
""'" anddliel.en. delenniaaIioa mccbaIIiom of illSed..
Fie:. 29: Sex determination systems

Though in all organisms sex determination depends on one principal,
but construction of chromosome is different in different animals. In
human, animals and Drosophila and other insects female have XX
sex chromosomes and male have XY sex hromosomes. In some animals
Y chromosome is absent. On the hand in birds and moths female is
heteroxygous and male is homozygous. In other words female has
XY sex chromosome and male has XX sex chromosome.
In some insects, female has only one X chromosome and Y is absent.
Therefore the female has XO chromosomes and male has XX
chromosomes as shown in the following table
118
Sex Detenninatioll
Types of sex chromosomes in various organisms

Organism Heteroxygous Sex Gametes Sperm Zygote Egg
Human, animals Male XY XY
& drosophila etc
Grasshopper & Male XO xx
Parotiner bug
Birds & moths Female xx XY
Fumeamoth Female XX XO
Sexual functions of X and Y chromosomes: X and Y chromosomes

though are called sexual chromosomes but they not only do the work
of sex determination because there are also genes which are
unconcerned with sex. In XY type of sex determination it is understood
that on Y chromosome there is no active gene for sex. Morgan on the
basis of researches on Drosophila melanogaster concluded that X
chromosome is free from gene. Bridges (1916) has seen a male in
Drosophila melanogaster where Y chromosome was absent. Despite its
size, shape and behaviour such fly was perfectly normal. But these
were sterile. Not only this, a female has been seen in Drosophila
where in addition to xx chromosomes there was one Y chromosome.
But such a fly was perfectly normal in size, shape, behaviour and
fertility. Therefore from :these facts the conclusion can be drawn that
in Drosophila melanogaster with presence of one X chromosome males
develop and with presence of xx chromosomes female flies develop.
(1 c) Genic balance theory:

Genic balance theory of sex determination was presented by Bridges
in 1921. He explained that in any sex determination sex chromosome
and body (somatic) chromosomes i.e. autosomes both works. X
chromosome is tempted to produce female. Whereas autosomes is
tempted to produce male. But all female determiners are not present
on X chromosomes. Similarly all determiners for male are not present
on Y chromosome. Both the type of determiners are infact present on
X as well as autosome chromosomes. However female determiners
are in dominant phase on X chromosome and male determiners are
in dominant phase onautosomes. Therefore if compared to autosome
the quantum of X chromosomes increases then sex determination goes
to femaleness. Similarly if quantum of autosome is more than X
chromosome then in that case sex determination is towards maleness.
Therefore it is the balance between X and autosome which determine
sex. In other words sex determination is dependent on ratio of XI A.
It has been explained in (Fig-30).
119
Diagrammatic representation of Genic balance theory in

Drosophila melanogast.
Fig. 30: Genic balance theory.

(2) Physiological theories:
(2 a) Metabolic differentiation theory:
Metabolic differentiation theory of sex differentiation was proposed
by Riddle. He expressed that male or female sex is dependent on
metabolic condition. According to him sex determination is not
dependent on chromosome differentiation. To prove it he gave
examples of sex reversal from male or female or vice versa.
(2 b) Quantitative theory of sex determination:
This theory was proposed by Goldsmith. Based on experiments he
stated that quantity of certain harmone determine sex of organism.
Quantity of harmone control metabolic condition. Sex determination
depends on. metabolic condition. Hence determination of sex depends
on quantity of harmone.
Sex reversion:
In human beings, animals, birds etc. sex reversion have been reported.
Explaining it Sharp (1934) mentioned that sex determination is not
dependent only on sex chromosome. According to him sex
chromosome is only a part of the system of sex determination. like
other inheritable characters sex development is also controlled by
interaction of gene and environment.
The geneticist Emersion stated that in dioecious plant and some animals
120
Sex DeteJ?tlination
genic balance is very delicate. Due to change in environment that

genic balance get disturbed resulting in sex reversal. Shaffner working
on Crotolaria juncea reported that growth of the plant in shallow soil
and deficient light result in sex reversion. He also found out that by
treatment of Crotolaria juncea plant with smoke of tobacco, sex
reversion takes place. In tomato (Lycopersicum esculentum) deficiency
in carbohydrate results no development of male sex and deficiency
in nitrogen result nO development of female sex. In case of castor
(Ricinus comunis) too much pruning results more development of
female sex organ. Edmonds ( 1930) reported thatin cucumber exposer
of the plants for sharp light for long duration increases growth of
male sex. On the other hand in reverse condition female sex grows
more.
Intersex and supersex in Drosophila:
Bridges (1922) working on Drosophila melanogaster reported such
females in which x chromosome and somatic chromosomes occurred
in triplicate. Such triploid female fly resembled to diploid female in
all respects including fertility.When this female was hybridized with
diplOid male, then 8 type of offsprings were produced.
(1) Triploid female: It has 3X chromosomes and 3 sets of autosomes.
(2) Normal diploid female: It has 2 X chromosomes and 2 sets of
autosomes.
a) Xxy female.
b) Intersex: It has 2 X chromosomes and 3 sets of autosomes
(Fig-31, 32).
c) Intersex having 2 Xchromosomes, 1 Y chromosome and 3
sets of autosomes.
d) Normal male: It has 1 X chromosome, 1 Y chromosome and
2 sets of autosomes.
e) Super male: It has IX chromosome, 1 Y chromosome and 3
sets of autosomes.
f) Super female: It has 3 X chromosomes and 2 sets of
autosomes.
Gynandromorph or Gynander:
Sometimes in animals some parts of body is male and other part is
female. Such animals are called Gynandromorph or Gynander.
Gynandromorph has been observed in Drosophila, silk worm and
121
Tnplold Diploid
3A+XXX ¥ 2A+XY 0
A+X I~
Ga~t:+y \
Tnplold female Tnplold Index

2A+XX 3A+XXX 3A+XXX
Diploid female Diploid male

A+X 2A+XX 2A+XY
2A+X Tnplold Intersex Super male

3A+XX 3A+XY
A+XX Super female Diploid female

2A+XXX 2A+XXY
Results obtamed from 2 Brtdge's classical cross of a triplOId (3A+XXX)

female fly and a diploid (2A+XY) male fly (Drosophila)
Fig. 31: Effect on sex of the balance between X chromosomes and

autosomes in Drosophila melanogaster as observed by Bridges (1922).
Number of Number of Sexmdex=

Phenotypes chromosomes autosomes No X's
(A sets) No. A sets
Super female 3 2 15
} letraploid 4 4 10
Normal triplOid 3 3 1.0
female diplOid 2 2 1.0
haplOid I I 10
thts X chromosome
WIll be lost Intersex 2 3 067
Nonnal male I 2 050
Super male I 3 033
The loss of an X chromosome dunng mitosIs
10 a 2A+XXcell and formatton of two types Different doses of X-chromosomes and autosome sets and thetr effect on sex determmatlon In
of cells -XX and XO DrosophIla
Fig.32: Intersex in Drosophila melanogaster.

honey bees. Morgan and Bridges (1919) on the basis of chromosomal
theory described gynandromorphy and told that these are produced
by zygote having two x chromosomes in Drosophila. Individual with
2X chromosomes are female, therefore gynandromorphy develop first
as female. But sometimes due to irregular cell division X chromosome
gets destroyed. Therefore all the resultant cells have only one sex
chromosome. Organism with one x chromosome are male. Organism
with two x chromosomes are female. During occurrence of misdivision
122
Sex Determination
of zygote one X chromosome disappear. Therefore half part of

gynandromorph is male and other half part is female (Fig-33). But it
is not necessary that division of zygote is misdivision. It is also
possible that after the individual is half grown if at that time
misdivision of cell occurs then 1/4 part of the body become male and 3/4
of the body becomes female. It is also possible that when growth of
the organism is almost complete then misdivision of cell may happen
deleting one X chromosome. In that case only a small part of the
body shows male like symptom.
A biIIIetaI i)IlIIItdt;OInorp/1
D~14.
Fig.33: A gynandromorph of Drosophila melanogaster - the half side of

the fly is female and other side is male.
Goldsmidt and Katsuki working on sex determination of silkworm
found out another mechanism of development of gynandromorph.
They could observe such egg cell which had two nucleus. If one nucleus
is fertilized with a sperm having X chromosome and the other nucleus
is fertilized with a sperm having y chromosome. In that case the
zygote with xx chromosomes are female and the zygote with XY
chromosomes are male.
Kinds of gynandromorphy:
According to presence of sex tissue type gynandromorphys are of
following types.
(1) Bilateral gynandromorph: Half of body is male and other half is
female.
(2) Anterio-posterio gynandromorph: Front portion of the body is
one sex and back portion of the body is another sex.
(3) Sex piebalds: In the body of female there are patches of male.
123
Importance of sex determination

Horticulture, crop culture and poultry have lot of importance of sex
determination. Several fruits and crops are monosexual and also
bisexual. It is important to identify the female and male sexes.
Similarly in poultry it is also essential to identify male and female at
the time of birth.
124
Chapter 16
Sex Linked Inheritance
It has been explained in the chapter on sex determination that sex
chromosome play a great role in heredity. Experiment by Morgan
and Bridges have proved that sex chromosomes have special genes
which are not found in autosomes (somatic chromosomes). By
observations of Wilson it has been clear that both the heterogamic
sex chromosomes x and y differs in structure and types of genes.
Various genes which are present in sex chromosomes X and Y are
known as sex linked genes. The characters governed by the sex linked
genes are called sex linked characters. Trasmission of such sex linked
characters from one generation to another is known as sex linked
inheritance.
Structure of sex chromosomes:
There is enough difference between structure of X and Y chromosomes.
Y chromosome is smaller in size. X chromosome is rod like and
straight. Y chromosome is slightly bent on one end. It has been also
observed that x and Y chromosomes have unequal arms. The small
arm of X chromosome matches with one arm of Y chromosome. Other
two arms are dissimilar (Fig-34) and nonhomologous. It has been
seen that in non homologous parts of X and Y chromosomes there are
incomplimentary genes. On the other hand on homologous parts
there are complimentary genes. During meiosis non homologous parts
never do pairing and never have crossing over. In this way the genes
on non homologous parts are completely linked. On the other hand
homologous parts do have pairing at synopsis stage of meiosis and
also form chiasma and crossing over. Therefore genes of homologous
parts are incompletely linked.
Kinds of sex linked inheritance:
Inheritance of sex linked genes is different from inheritance of ordinary
genes. Non homologous parts behave in different ways. Sex linked
inheritance are of following three types.
125
(1) Digenic sex linked inheritance.

(2) Holandric sex linked inheritance.
(3) Hologenic sex linked inheritance.
(4) Diandric sex linked inheritance.
These 3 sex liked inheritance are described below:
(1) Digenic sex linked inheritance:
Digenic sex linked inheritance is those genes which are present in
nonhomologous parts of completely linked genes and behave like
ordinary genes. These genes are passed on in offsprings through the
female. There never go directly from father to offsprings. Such
inheritance is called digenic inheritance and such genes are known as
digenic genes.
.._--
__--
.....
SMCY _ __
ElF1AY r_ _ _
IIIII 11 tllIl~lIllH [ 1 1 T
._--
x
-- --. -
-,
'--- Comparing the X and Y Diseases
_._x_
TheX __
---- -- ""----
I ........-
x
-----.
:-
- ___
-:.'i55"'"
,~\I!IO~,Q!lO
_ _ Il0010'/
_ _ whoM_
d _ Defeetlve_on
:i---
.............
IheX~_Io_1n _ _'t
------------.---- -----
-..... " .... ,.
y .
10---.
---
r- .......•
Fig. 34: Structure of human sex chromosomes.

126
(2) Holandric sex linked inheritance:

These are those genes which are present in non homologous parts of
Y chromosome. These genes are completely linked to Y chromosome.
Those genes are directly transmitted from father to offsprings. This
type of inheritance is called holandric inheritance and such genes are
known as holandric genes.
(3) Hologenic sex linked:
The two X chromosomes of female behave in such a close way that
these chromosomes are completely homologous. One X chromosome
of female passes on directly to daughters. Such inheritance is called
hologenic sex linked inheritance and such genes are called hologenic
genes.
(4) Diandric sex linked inheritance:
If the X chromosome from mother first passes on to sons and then it
get transferred to daughters, then such a inheritance is called diandric
sex linked inheritance and such genes are called diandric genes.
Sex linked inheritance in Drosophila:
Based on structure of sex chromosomes Drosophila melanogaster and
human being both have XX in female and XY in male. In this fly Morgan
for the first time studied sex linked inheritance. When Morgan was
rearing cultures of red eyed Drosophila at that time he saw white eyed
fly. First he thought it to be a mutant. But when Morgan hybridized
this white eyed male Drosophila with red eyed female Drosophila he
observed that all the offsprings (males as well as females) has red
eye (designated by the letter W). Morgan concluded that W for red
eye was dominant over w which was responsible for white eye. When
male and female of Fl red eyed Drosophila were hybridized then in
the F2 he found 3 red eyed and 1 white eyed Drosophila. But
noteworthy matter was that all the white eyed Drosophila were male
only. Red eyed Drosophila were also present in equal number.
The most astonishing matter was that all the females had red eyed
only. From this observation Morgan concluded the w gene responsible
for white eye was located in X chromosome of male Drosophila and
w responsible for white eye expressed only in absence of W gene.
In the above mentioned example the gene w is first transmitted into
daughters. Afterwards it get transferred in sons. To verify these
results Morgan hybridized Fl heterozygous red eyed females with
127
hemizygous or homozygous white eyed male parent (test cross). He

observed that in female Drosophila half number was with white eye
and in male Drosophila half number was with red eye.
Sex linked inheritance in human
One good example of sex linked inheritance in human being is color
blindness. Color blindness is of several types and in different grades.
The most common of color blindness is the inability to differentiate
between red and green. It is known as 'red-green' color blindness. If
a person is unable to identify red color then technically it is called
'protonopia' If a person is unable to identify green color then it is
technically called as 'deuteranopia'. Protonopia and deuteranopia both
are hereditary. The responsible genes are recessive and sex linked.
Following facts have been observed in this regard:
(1) Women having normal eye whose father were a color blind, when
married with a normal eyed male produces children of who half
have normal eyes and other half have color blind eyes.
(2) Color blind women produce color blind sons. If husband of such
a women is color blind then in the offsprings only the girls will
be color blind.
(3) Generally father of a color blind women is also a color blind.
(4) Color blindness disease is more common in gents as compaired
to ladies.
In USA color blindness is found in 8% gents and 0.5% ladies. Why
this disease is more common in gents? The answer of this question
can be found if we imagine that the gene responsible for color
blindness is present on X chromosome and the heterogametic sex XY
is male.
If a normal eyed man marries a color blind women, in that case XX
chromosome of that man will be transferred to his daughters only, it
does not reach to sons at all. From that woman out of two X
chromosomes one XX goes to daughter and the other goes to son.
Because the gene for color blindness is recessive to the gene
responsible for normal eye. In man (male) there is only one XX
chromosome which comes from color blind mother. Therefore all
their sons will be color blind. In daughters there are two XX
chromosomes, out of which one come from father and the other come
from mother. Because one XX chromosome from father there is gene
for normal eye which is dominant therefore all daughters of F1 have
normal vision. In these daughters the gene for color blindness is in
128
.recessive form. Therefore if these girls are married with man having
normal vision, then all the female offsprings will have normal eye
but half of the male offsprings will have normal eyes and other half
male offsprings will have color blind eyes. When color blind man
marries..- women who have homozygous or heterozygous color
blindness-in that case they will produce color blind female offsprings.
But such marriages generally do not take place. This inheritance is
fully dependent on X chromosome. It can be also understood by Fig-
35.
Fig.35: Inheritance of color blindness.

Inheritance of sex linked characters through Y chromosome:
We have understood before that the sex linked genes present on X
chromosome, do not have any allele on Y chromosome. This is the
reason that when the recessive gene is alone in male then only male
express itself. Compared to X chromosome Y chromosome is rather
inert. In other words the genes of X chromosome do not have allele
on Y chromosome. But in Drosophila melanogaster there is bobbed gene.
This gene has allele on X as well as on Y chromosome. Due to bobbed
gene the Drosophila produces normal bristle. The mutant gene which
is recessive produces slender bristle. When in male X chromosome
has mutant of bobbed gene and Y chromosome has bobbed gene then
the offspring produces normal bristles. If a male having such normal
bristle, is crossed with a female having bobbed mutant genes on both
129
Genetics : Fundamentals and Applica tions
X chromosomes then all the male offspring will have norma l bristles
.
But all the females will have slende r bristle. In other words norma
l
bristle is transm itted from father to male offsprings. Therefore we
can say that inheri tance of bobbe d charac ter is depen dent on Y
chrom osome . Anoth er examp le of audito ry condit ion can be well
unders tood by Fig-36.
Father
Wrth
Audmry Hearioa
Condition Mother
Xy Xy
Oa\llhtcr
Wuh
?J
H_int
S-
Da..qht«
Wath
~
H-int
Son
Audtory Auditor,
Condition Coadlion
Fig.36: Inheritance of sex linked character through Y chromosome.

Sex influe nced characters:
There are some genes whose two alleles express domin ant effect in
male and female sexes. In other words the allele which is domin ant
in male, the same allele expresses recessive in female. Similarly the
allele which is domin ant in female the same allele expresses recessive
in male. Such genes are called sex influenced characters or genes.
Sex limite d character:

There are certain characters which are expressed in one sex only. The
governing genes are presen t on autosomes. Such characters are called
secondary sexual characters or sex limited characters. Its inheritance
is called sex limited inheritance. Appearance or no appearance of such
characters depen ds on sex hormo ne which is produc ed in ovary or
testis.
130
Chapter 17
The Gene
From Mendel time importance of gene increased tremendously. In
chapters so for, on every step we were concerned with gene. In
genetics every action is dependent on gene. Physical nature of gene
and its chemical structure were not understood well till recently. In
modern times genes have same place in science which was had earlier
by hormones and vitamins. Now we very well know where the genes
are located, how they express themselves and what is their physical
and chemical nature. Geneticists, cytologists and biochemists all over
the world are busy in analysis and trying for artificial production of
genes. After artificial synthesis of genes many difficulties of human
beings will disappear. Then man can manipulate so many important
affairs such as mental capabilities, sex power, excellent health etc. He
will be able to produce more desirable varieties ofplants and animals.
Detail information about genes are given below:
Definition of gene:
Based on studies of Watson and Crick (1958) and Wilkins (1962) genes
can be defined as mentioned below:
Gene is a molecule or a chemical radicle made of carbon, oxygen,
hydrogen, phosphorus, nitrogen and deoxyribonucleic acid which are
attached on a string chromonema made of protein. Genes are
transmitted from one cell to another, from one generation to another
without any change in their shape and structure.
Physical nature of gene:
Complete information on physical nature of gene is still not available.
Whatever knowledge available on that basis it will be explained that
where genes are located and what is their shape and size:
(a) Position of genes:
We all know that genes are located on chromosomes. But in
chromosome where are the genes located? Answer to this question
was first presented by Demerec (1949). He explained that most
131
important part of chromosome is chromonema, which is in form of

thin string in full length of the chromosome. It is homogenous in the
whole length and uniform in chemical and physical nature. Genes are
located on this chromonema. Genes differ from each other chemically.
(b) Shape of gene:
Gene could not be seen so for. So regarding its shape nothing can be
mentioned. However by electron microphotographic technique it could
be now possible to through light on its shape. In this respect work
done by Slizynski (1952) explained that on salivary gland chromosome
and work done by Stanley (1952) is on tobacco mosaic virus. Slizynski
(1952) explained that on salivary gland chromosome of Drosophila
melanogaster there are numerous dark coloured bands which are
probably genes. Stanley (1952) expressed that virus looks like a rod
and behave similar to gene in all fundamentals. Based on conclusions
drawn by these scientists genes are very minute and rod likes
structures. This idea is supported by x ray induced mutation's target
theory. In addition to it, this also get support from a observation that
if gene has dot like structure then there will be much less surface for
hitting with radiation. But it is a fact that with hit by a small dose of
radiation there is a change in gene. Therefore it is clear that shape of
gene is not like a dot but it is small rod like.
For determining the shape of gene the most important work was the
work of Watson and Crick (1958) and Wilkins (1062). They have
presented the virtual shape of gene. Ac!=ording to these scientists
gene's shape is spiral where two rod like structures overlaps on each
other spirally in a helix shape.
Size of gene:
To find out size of gene is a very difficult work. Electron microscope
has helped to'some extent in this task. Butto see by electron microscope
the genetic material has to be treated with several chemicals. Due to
these treatments the gene undergoes changes. The result is that it
becomes extremely difficult or almost impossible to find out exact
size of gene. Therefore beside electron microscope ionizing radiation
are used for this purpose. After hitting chromosome by ionizing
radiation (particles) mutations are measured and size of gene is
computed. But this technique is not fool proof. Therefore scientists
have put forth another technique. In this technique first number of
genes is found in a chromosome. Afterwards by calculation size of
.gene is found out. Though this technique also has shortcOmings, even
132
The Gene
then possible size of gene can be presented which can be compared

by virus. According to geneticists gene is a macromolecule which
measures between 15 mu x 100 mu and 20 m u x 150 mu .
Chemistry of gene structure:
Gene is made of 5 elements viz. carbon, hydrogen, oxygen,
phosphorous and nitrogen. These elements unite and form nucleic
acid. Nucleic acids are of two types viz. 1- DNA (Deoxy-ribonucleic
acid) which is present in nucleus. 2- RNA (Ribo nucleic acid) which is
mostly present in cytoplasm. DNA and RNA both are made of
numerous nucleotides. These nucleotides are arranged well on each
strand of helix. Each nucleotide is made of a phosphate group and a
nucleoside. It is a complex molecule.
Phosphate group is a derivative of phosphoric acid (H3 P04). A
nucleotide is of primary phosphate type. If two or more nucleotide
unites then it become secondary phosphate type. The difference
between these two types of phosphate group is in number of hydrogen
atoms
Nucleoside is a chemical unit like nucleotide which is made of sugar
and a nitrogenous organic base. Sugar by which nucleoside are made
of have pentose nature like glucose. In that there are 5 atoms of carbon
DNA
Fig. 37: Double helical structure of DNA (gene)
133
in a chain. These sugars differ from DNA and RNA nucleoside. In

RNA it is ribose and in DNA it is deoxyribose. Because the nucleoside
obtained from nucleus is deoxyribose therefore this nucleic acid is
called deoxyribose nucleic acid or DNA. DNA is considered to be a
gene (Fig-37).
Sugar may be ribose or deoxyribose made of 5 carbon atoms. On first
and fourth carbons there are a independent and a solitary oxygen
atoms. In the structure there are hydrogen atoms and hydroxyl groups
(OH). In ribose sugar there are 6 hydrogen atoms. In deoxyribose
sugar there are 7 hydrogen atoms. Another difference is that in ribose
sugar there are 4 hydroxyl group, but in deoxyribose sugar there are
3 hydroxyl (OH) group (Fig-38 ).
Fig. 38: Chemical structure of ribose and deoxyribose.

Nitrogenous base are ring like organic compound. These are of two
kinds ego 1) Single ringed pyrimidines and 2) double ringed purines.
Pyrimidine are of three types ego Cytosine, Thymine and Uracil.
Purines are of 2 types e.g. Adenine and Guanine.
In DNA there are adenine, guanine, cytosine and thiamine. But in
RNA there are adenine, guanine, cytosine and uracil. In every
nucleotide sugar is joined at one side with phosphate and at other
side with purine or pyrimidine base ( Fig-39).
Following are the main features for chemical nature of nucleic acid
or genes (Fig-40):
(1) Nucleosides of both the strands of helix are joined together with
2 or 3 unstable hydrogen bonds.
134
The Gene
Thymine
(In DNA only) Cytosine
H
UracH (In ANA only)
Fig. 39: Structure of purine and pyrimidine base.
(2) Pyrimidine nucleotides of one strand of helix are joined with

purine nucleotides of other strand of helix.
(3) Adenine base (purine) of a nucleotide are joined with thymine
base (pyrimidine) of opposite nucleotide in DNA, but in RNA
it is joined with uracil base.
(4) In DNA and RNA guanine base (purine) of one strand are joined
with cytosine base of other strand .
(5) Nucleotides are arranged one behind the other in straight line.
Thus it confirms linear arrangement of genes.
(6) Nucleotides of strands of helix are joined to each other by sugars.
(7) On each strand of helix nucleotide are arranged in sets of 3 viz.
adenine- cytosine - guanine. The set of 3 nucleotide is known as
triplet or trinucleotide
In DNA molecule the exact number of nucleotides and actual sequence
of triplets in gene are yet not clearly known. In addition to it exact
length of helix is yet to be known because in the process of extraction
helix gets broken. However based on available records number of
nucleotide pairs is around 300000 and molecular weight of nucleotide
is about 300. It has been calculated that difference between nucleotides
is 3 to 4 Angstron units. Length of DNA molecule is about 60000000
or 6 x 106 Angstron units. Sequence of nucleotide in DNA is called
genetic code.
135
Fig. 40: DNA double hehix.

Sub units of gene:
Gene is made of following sub units:
1. Cistron:
Benzer (1962) named the largest segment of gene as Cistron. This
name is based on that segment of DNA which has polypeptide chain.
In Cistron there is a long chain of nucleotides. It is capable of mutation.
Mutation and recombination takes place in Cistron.
2. Muton:
According to Benzer (1962) mutton is that smallest part or Cistron
which is capable of mutation. Experiments on bacteria and several
proteins have indicated that muton can be as small as a nucleotide.
3. Recon:
The third subdivision of gene is recon ( Benzer, 1962) Recon is the
136
The Gene
smallest unit of DNA which is capable of crossing over and

recombination. A recon may be as small as a nucleotide of DNA or
RNA.
Properties of gene:
Following properties of genes have been found out by a lot of research
work.
(1) Physical nature of gene is similar to virus.
(2) Except mutated genes all other genes are useful to the organisms.
Some mutated genes are also useful.
(3) Gene has the capacity to replicate to its own type.
(4) Any gene can mutate.
(5) Every gene is made of molecules of sugar, phosphate, purine
bases (adenine and guanine) and pyrimidine bases ( cytosine
and thiamine).
(6) Gene is made of numerous nucleotides.
(7) Every gene has the capacity of crossing over and recombination.
(8) In chromosomes genes are arranged in linear fashion.
(9) In one cell division genes replicate only once. Every gene controls
a type of enzyme production. The enzyme controls some
physiological and metabolic functions.
(10) Every gene produces m RNA and sends it to cytoplasm. The m
messenger control synthesis of protein.
(11) Process of gene replication occurs only in living cells.
(12) Every gene is made of 3 sub units viz. cistron, muton and recon.
(13) Very closely located genes exhibit linkage.
Chemistry of gene mutation:
Till chemical structure of gene was not known, the process of gene
mutation was only an imagination. But now it is clear that due to
mutation there is some change in the sequence of nucleotide- it is
infact gene mutation. For example if in a gene the sequence of
nucleotides is ATCTGA / TAGACT. If sequence of a nucleotide gets
changed due to mutagen then the character and effect of the gene
also get changed. Therefore on this basis it can be said that a change
in sequence of nucleotide is mutation.
137
Genetics: FUlldamentals and Applications
Action of physical mutagen on gene:

Among all physical mutagen the heat (through sun light) is an
important mutant because due to increase.in temperature in nucleotide
pairs, the hydrogen bond of purine and pyrimidine bases become
week. Because of weekness of hydrogen bQnds in nucleotides iL place
of adenine the guanine comes and in place of cytosine the thymine
comes. This is also a type of mutation. Ultraviolet (UV) rays may also
cause mutation.
In case of mutation induced by radiant energy, in cells, free water
molecules are produced which when collides with nucleotides,
hydrolyses the bond between bases. In addition to it sequence of
nucleotides get changed. In other words mutation occur (Fig-41).
Fig.41: Action of ultraviolet rays on gene in producing mutation.

Action of chemical mutagen on gene:
For induction of mutation several chemicals have been discovered.
Mustard gas is quite effective. In addition to it Methyl Ethane
Sulionate, Ethyl Methane Sulionate, 5 Br-Uracil and HNO 2 are
quite important.
Action of HNO 2 on gene:
Action of HNO 2 on gene has been explained by Fig-42. With effect
of HNO 2 adenine gets changed to Guanine and Cytosine gets
changed to Uracil or Thiamine. Due to these changes the sequence of
nucleotides also get changed. So this becomes mutation.
Action of 5 Br-Uracil:
The process of gene mutation production by 5 Br-uracil can be well
understood by (Fig-43). If a gene has nucleotide sequence of ABCT /
TCGA. When 5 Br-uracil acts on this gene then this chemical mutagen
come in contact with nucleotides ACGT. This DNA (gene) during
replication breaks off into two strands AGCT and TCGA. Because 5
138
The Gene
-"-
('(--7'-
".My"
o
......... - ...
-..
Fig. 42: Action of UNO 2 on gene in producing mutation.
It) PfIt... .,,~lICII ..

11
II" ......'
/
'I\ O""---H-H
1/ \ C-C /1'101
H::::,...
c-c I
IfS • )\N - t l - - - Hq'
tIC, I
S,\
4C- \
'N
\ I 2/ \ 2 1/
H--C fIC=N
/ '\
~ ;Won""
1_...... e'" tUle'
lbl paitloI&otS_""l/wilh..-....:
ftP\ImioII
"iX:sita&l
_plait)
Fig. 43: Action of 5 Br-uracil on gene in producing mutation.
139
Br-uracil ( 5 BU) is just like thymine. Therefore 5 Br-uracil get replaced

in place of T ( Thymine). Therefore after replication along with TCGA
strand there is a modified strain AGC 5 Br-uracil. Similarly TCGA
becomes 5 Br-uracil CGA. Therefore from old DNA the two new
DNA will be AGCT / AGe 5 Br-uracil and TCGA / 5 Br-uracil CGA.
This change in nucleotide is the mutation. During second replication
5 Br-uracil (5 BU) replace Guanine (G) instead of Thiamine (T).
140
Chapter 18
The Gene Complex
It is clear that every character of organism is affected by gene. Till

Mendel's time it was believed that every character was controlled
separately by individual gene. Afterwards the researches of several
geneticists (e.g. Bateson, Punnet, Shull, Nelson, Ehle etc.) have proved
that every trait (character) is controlled by combined effect of more
than one gene. Scientists have also observed that different genes
control characters differently. Sometimes one gene controls more than
one character and sometimes 2 or 3 genes control only one character.
These facts reveal that genes behaviour is quite complicated. Therefore
studies of gene behavior is known as gene complex.
According to behavior, genes can be classified as mentioned below:
. (1) Basic genes
(2) Lethal genes
(3) Multiple genes
(4) Cumulative genes
(5) Pleiotropic gene
(6) Modifying gene
These are described below:
(1) Basic genes:
Basic genes are most common. These are of following types:
(A) Normal gene: Single pair of gene which express one
character is called normal gene. For example in rat gene CC
for skin colour.
(B) Complimentary gene:Genes which independently have
certain effect, but together the genes express different effect.
Such genes are called complimentary genes.
(2) Lethal gene:
The gene which has effect of killing the organism is known as lethal
gene. Lethal gene generally expresses their effect at embryonic stage.
141
Some times lethal gene expresses their effect later on.

Lethal genes are found in almost all types of plants and animals.
However they express only in homozygous phase. When they are in
heterozygous phase then development of individual is normal and
presence of lethal genes is not noticed. But the organism having lethal
gene in heterozygous condition when produce offsprings then those
offsprings die which get lethal gene in homozygous condition.
Example of lethal gene:
Lethal gene can be easily understood by examples of rats. When
hybridization is done between such rats which express yellow color
in heterozygous condition. Then the offsprings do not show the ratio
of 3:1, but the offsprings show a ratio of 2:1. The reason for this
abnormality is presence of lethal gene.
Suppose yellow heterozygous rats have the genotype Yy, then at
gamete formation Y and y genes will go to different gametes. Now if
Y and y sperms of male rat join with Y and y egg cells of female rat
then the offsprings will be of 4 types. But the offspring having YY in
homozygous condition will die. This type of lethal effect has been
observed in several organisms.
(3) Multiple genes:
Some times two or more pairs of genes interact and produce a
phenotype. Such genes are called multiple gene. These are of following
two types:
(A) Duplicate genes: If two pairs of genes control a phenotype
then these are called duplicate genes.
(B) Triplicate genes: Triplicate genes also have effect like
duplicate genes. The difference is that instead of two pairs
there are three pairs of genes.
(4) Cumulative genes:
Due to cumulative action of dominant genes expression of characters
is intensified. Such genes are called cumulative genes.
(5) Pleiotropic gene:
Gene which expresses more than one character at a time is called
pleiotropic gene.
142
The Gene Complex
Example of pleiotropic gene:

In house rat the gene dw not only causes dwarfness but also causes
other characters such as sterility, improper development of thyrOid
gland and absence of secondary sexual characters, The reason is that
dw gene control harmone development by pituitary gland. Because
pituitary gland controls almost every part of body. Therefore if there
is some disturbance in pituitary gland that causes change in tissues,
organs and systems. Infact dw gene stops the growth of eosinophilous
cells which forms harmone in the gland. Disturbance in pituitary gland
stops or effects the secretion by gonadotrotin, thyropin and stops
the development of growth harmone. Stopage of these harmones
results sterile condition of gonads, defective growth or dwarfness.
In rats having dw gene which inhibits function of pituitary gland, if
by surgery a functional pituitary gland is implanted then the growth,
fertility etc become normal.
(6) Modifying gene:
Those genes which do not produce an effect themselves but has
capacity to modify the,. character are called modifying genes. These
genes are of two following types:
(A) Supplimentary genes: These genes do not produce any effect
but by interaction with other genes intensify or dilute the
expression of character.
(B) Inhibiting genes: Inhibiting genes do not produce any effect
itself but interact with other genes to prevent or reduce the
action of other genes. Inhibitary gene is also called epistatic
gene. It has been presented in detail in the chapter laws of
inheritance.
143
Chapter 19
Gene Action
Characters of organism express because of cumulative effect of some
chemical action. It is now clear that genes responsible for one character
do not act at one time. However they come in action one after the
other. Each gene produces enzyme. Enzymes are produced by
autocatalysis activity with the chemical substance present in cell. One
enzyme performs only one chemical action which is in fact required
for activation with second enzyme. On changes brought by second
enzyme, the third enzyme exerts its effect. That is how this activity
continues on. In this chain of reactions if there happens any mistake
by any enzyme then the whole chain gets disturbed. For example if
enzyme a, b, c act on substance A and changes it in to the substance
D. If the enzyme b is not produced due to any reason then as explained
below only the substance B will be formed.
Gene a gene b gene c
Enzyme a' enzyme b' enzyme c'
I A._______B C.________D
11 A _a'___c' B
(no more changes afterwards because b' enzyme is absent)
The above mentioned diagram clearly express that any enzyme acts
only when there is proper background. For example the enzyme c'
will act only when the substance C is available. One more fact is clear
from this diagram that indirect effect of gene also produce character.
Some important actions of genes are mentioned below:
(A) Genes and pigment: Before understanding of gene action it is
important to know about pigment. In different organisms different
pigments are produced. Knowledge has been available about
chemistry of pigment production.
We will discuss production of pigment and color production in some
plants and animals as mentioned below:
144
Gene Action
Genes and flower color:

In flowers pigments are produced generally by anthocyanins &
anthoxanthins and their chemical derivatives. Both of these pigments
are dissolved incell sap. Anthocyanin produces various colours such
as red and blue and also pink, purple, magenta & lavender.
Anthoxanthins produces yellow and ivory colours in flowers.
Anthocyanin and anthoxanthin are made of one type of ring structures
which are derivative of sugar.
Anthocyanin pigment's one or two molecules are of sugar those are
joined with phenyl ring - peragonidin. In phenyl ring of peragonidin
on fourth carbon there is one OH group. It produces scarlet colour.
By the increase of OH (hydroxyl group) in phenyl ring the colour
changes from red to blue. In Delphindin there are 3 OH groups.
Therefore it produces blue black or violet color. Researches have
revealed that number of OH group is controlled by special genes.
In this connection one thing we should know that anthocyanin in
acidic medium is red colour and in alkaline medium it become blue
color. Therefore by growth in acidic and alkaline medium redness
and blueness increases respectively.
Several geneticists such as Lawrence, Seottt Moucrieff, Sturgess and
Wheldale on basis of several experiments have proved that in flowers
several colours such as red, blue, yellow, ivory etc. are produced by
action of special genes. These actions can be of several types for
example number of hydroxyl group, oxidation & reduction action on
third, fourth & fifth carbon atoms on phenyl ring and acidity &
alkalinity of cell sap or in other words change in pH. Actions of genes
can be well understood by following examples:
Example-l
In case of flowers of Streptocarpus due to actions of several genes
seven different colours are produced. Streptocarpus rexii flowers are
of blue colour. Streptocarpus dunnii flowers are of red colour. In these
flowers anthocyanin pigment is produced by gene A. Flowers having
aa gene do not produce anthocyanin so the flowers attain ivory
(white) colour. In Pelargonidin the pigment anthocyanin is produced
in absence of R and 0 genes. In Cyanidin anthocyanin is produced in
presence of gene R. In Delphinidin it is produced in presence of gene
O. In presence of gene R there is increase of one OH or CH 2 group in
phenyl ring. It makes the colour as rose or magneta. Due to presence
of gene 0 there is increase of CH 2 and OH groups. With the result
145
the colour changes to blue or mauve. In addition to it gene action

takes place through substitution of molecules also. For example on
third or fifth position of carbon in anthocyanin molecule, a molecule
of hexose get substituted. In a molecule of gene dd hexose and pentose
are located on third carbon atom and at fifth carbon atom there is no
substitution. This type of changes produces blue and salmon color.
Example-2
In Dahalia variabilis plant gene B produces anthocyanin and gene I
produces opigen (a type of anthxanthin). It may happen that due to
polyploidy number of genes B and I increases. Accordingly intensity
of colour gets affected as mentioned below.
bbbbiiii = No pigment (white)
Bbbbiiii = Cyanin (chocklate)
bbbbIIii = Epigen (ivory)
BbbbIIii = Cyanin & Epigen (purple)
BbbbIIIi = Cyanin, Epigen (carmine) & Pelargonidin (magneta)
BBbbIIii = Cyanin (carmine) & Pelargonidin
Genes and pigments in animals:
Like in plants color (pigment) production in animals take place due
to gene action. In animals black color occurs due to melanin pigment.
Melanin pigment is produced by enzyme action on colorless
chromogen tyrosine (amino acid).
In Negro and black animals amont of melanin is more. Therefore
their colour is black. On the other hand in white animals and fair
colour persons the amount of melanin is less. In the above mentioned
figure oxidation of tyrosine to form dihydroxyphenylanine happens
because of dominant gene C. The same gene C produces the enzyme
tyrosinase. Whenever the gene c is in homozygous condition (cc) then
production of tyrosinase stops. Therefore melanin production also
stops. The result is albinism or white colour.
Wright on basis of experiment has sta"ted that in animals colour
development takes place due to action of two enzymes. In white as
well as black both animals chromogen is found in equal amount. Both
enzymes have capacity to oxidize the chomogen. But the process of
oxidation takes place in two different types.Wright has also stated
that if the enzyme Lis present then chromogen is oxidized into yellow
146
Gene Action
pigment but in addition to I if there is double dose i.e. 11 then 11

enteract with I and produce brown or black pigment. But if 11 are
alone then no pigment is formed. Appearance of different colors
depends on quantity of both enzymes as mentioned below:
(1) Chromogen (equal number of Enzyme 11 and Enzyme I) >Black
(2) Chromogen (unequal amount of Enzyme I & 11) >Black
(3) Chromogen (enzyme I half quantity) >Red
(4) Chromogen (enzyme 11) >No colour (white)
Gene and eye color:
For appearance of different colours in eyes of insects gene action was
explained by Kuhn, Beadle and Ephrussi based on their experiments.
In Drosophila melanogaster and caterpillar of Ephastia, eye colour and
body colour depends on the presence of Ommochrome pigment. This
pigmentis produced from amino acid tryptophan by action of enzymes
produced by some genes. In these organisms there are two genes V
and en which produce two types of enzymes V' or Tryptophane pyrholase
and enzyme Cn'. By the action of enzyme V' on tryptophan one amino
acid Kynurenine is produced. On this the enzyme Cn acts and changes
it to 3-hydroxyl amine. This gets changed into amino acid
ommochrome which in presence of protien carrier provide brown
color to eyes and body (Fig-44).
In the above mentioned Fig-44, if gene V by spontaneous mutation is
changed to v then the insect looses its capacity to produce Kynurenine.
In such a situation if extract from V gene insect is injected in the body
of v gene insect, then again that insect start production of normal
pigment. Instead of injection the geneticist compare transplanted testes
of W or Vv insect in that mutated insect. The result was that the
mutated insect again started production of normal pigment.
Afterwords Kynurenine injection was found to be successful.
Similarly it has been observed that if Cn gene changes to cn by
mutation then the insect becomes unable to produce 3-hydroxyl
Kynurine from KynureRine. Therefore the insect is unable to produce
that wild type of pigment. But if testes of Cn Cn insect is transplanted
in the cn cn (mutated) insect. Then this mutated insect starts
production of wild eye color.
147
~H
? -C-c:ooH
~ I .. ) H .I.t,
-
•
~
I
I
I
I
•
. : :.
~ j
H H
((C:~-<:-COOH
H
H ......
N-<:-tI
"g
............ _ .-
....
,•
I
I
I
I
CQ-<:-<:-cooH
poI
::".
o "H,
I H I
IItt,
t_
H H
Fig.44: Formation of pigment from Tryptophan.

These observations suggest that in insects development of eye colour
depends on production of special enzymes which are effected by
special genes. But due to mutation these genes are unable to produce
the enzyme. The result is that the insect becomes unable to produce
the normal pigment. But if that desired enzyme is provided to the
mutated insect by injection or transplantation of testes then the
disturbed chemical reaction again starts and the insect start production
of normal pigment (color). In this connection the researches of Efrussi
and Beadle are worth mentioning. These geneticists have transplanted
eye disk in developing embryo. Their observations are listed below:
(1) When eye disk of wild type fly was transplanted in Vermilion
type or Cinnabar type of flies then these flies developed wild
type eye colour. Body enzyme of Vermilion or Cinnabar fly had
no effect.
(2) When Vermilion fly eye disk was transplanted in wild type fly
then the fly developed wild type eye only.
(3) When Vermilion fly eye disk was transplanted in Cinnabar fly
then the fly developed cinnabar eye only.
(4) When Cinnabar fly eye disk was transplanted in Vermilion fly
then the fly developed Cinnabar eye.
From above mentioned observations Ephrussi and Beadle concluded
that there is some deficiency in eyes of Vermilion fly and Cinnabar
148
GfmeAction
fly which can be corrected by the cells of wild type fly. Beside these
one more suggestion they made that because by the effect of Cinnabar
eye disk the eye disk of Vermilion fly changes to Cinnabar eye disk
but vermilion eye disk do not change the Cinnabar eye to vermilion
eye. Therefore it suggests that perhaps vermilion eye has some type
of deficiency.
The observations of Ephrussi and Beadle can be also understood by
biochemical facts. In case of Vermilion eyes enzyme V and Cn both
are absent and the amino acid tryptophan is present. In Cinnabar
eyes the enzyme V is present but the enzyme Cn is absent. In Cinnabar
eyes Kynurenine is present. Any of the both eye disk when
transplanted in wild fly then because of presence of V and Cn enzymes
in that fly wild eyes are produced. When Cinnabar eye disc is
transplanted in Vermilion disc eyed fly then in that fly only cinnabar
eye is produced because in vermilion disc eye Cn enzyme is absent.
When vermilion eye disc is transplanted in cinnabar disc eyed fly,
then in that fly only cinnabar disc eye develops because in vermilion
the enzyme V is present.
Above discussions make it clear that vermilion colour is due to
trytophan, cinnabar colour is due to Kynurenine, and wild type colour
is due to sommochrome. These facts indicate that genes play a great
role in production of different colours.
Gene and protein:
Proteins are made of amino acids, complex and chained molecul~.
The amino acids are joined together by peptide linkages. In various
varieties of a species proteins are of different types. Nature and
behavior of some special proteins such as enzymes and haemoglobin
are controlled by special genes. It can be understood by following
examples:
Example:
Sickle cell disease is caused by abnormal haemoglobin. Abnormal
haemoglobin is produced by difference in electrical charge in normal
haemoglobin. An intermediate stage has also been found when normal
and abnormal both type of haemoglobin coexist. These two types of
haemoglobin are produced because of difference in genes. Normally
si / si genes, abnormally Si / si genes and at intermediate stage Si / si
genes are found. During electrophoresis all the three types of
haemoglobins have movement at different speed because they have
149
different electrical charge. Normal haemoglobin has negative charge

(-), abnormal haemoglobin has positive charge (+) and intermediate
haemoglobin has negative and positive charge.
The geneticist Ingran has discovered that in normal and abnormal
haemoglobin one peptide linkage is different. From these facts it is
clear that when there is difference in gene action (gene si mutates to
Si) then one peptide linkage gets changed. This results such a defect
in haemoglobin that the person succumbs to death.
Genes and antigens:
Presence of different kinds of antigen in blood is because of genes. It
may be well understood that red blood cells (RBC) have antigen. By
this antigen during blood transfusion RBC reacts with antibodies of
donor blood serum. Every antibody and antigen has their own
chemical configuration. Land Stiener (1900) demonstrated that if blood
of incompatible blood group is transfused then RBC of the blood
clumps the RBC of the person. The result is that blood vessels and
capillaries of the person get blocked and therefore the person dies.
In human beings 4 types of antigen are found ego A,B,AB, and 0
(none). Persons of antigen A have such antibodies which if the blood
is in person of B antigen will agglutinate its red blood cells. Similarly
persons of B antigen have such antibodies which if the blood is
transfused in person of B antigen will agglutinate its red blood cells.
Persons with AB antigen do not have antibody. Persons with 0 antigen
have both the antibodies due to which A and B antigen red blood
cells get agglutinated. Experiments have revealed that these antigens
are produced by three genes viz. lA, IB and I as detailed below:
AB antigen - - > lA IB genes
B antigen - - >IB IB genes or IB I genes
A antigen - - > lA lA genes ,or lA I genes
o antigen - - > ii genes
Therefore,
Gene lA produces antigen A
Gene IB produces antigen B
Gene i produces antigen 0
Genes and human disease
150
Gene Action
Due to sufficient progress in biochemical genetics now it is clearly

known that several human diseases viz. albinism, alcaptonuria,
tyrocinosis, colour blindness, phenylketonuria etc. are produced by
action of genes. We will discuss some diseases below
1. Genes and albinism:
In human beings colour development is because of phenylalanines
melanine which is produced by amino acid - in subcutaneous cells.
Change of amino acid into melanin is controlled by genes K and C. If
due to mutation these genes are changed to k and c or due to any
reason the action of these genes are changed then formation of enzyme
is stopped. Therefore the amino acid - phenylalanine can not change
to melanin.
2. Genes, Tyrosinosis and Alcaptonuria:
In urine of some individuals p-hydroxy phenyl pyruvic acid is present.
Due to this the disease tyrosinosis develops. In urin of some persons
instead ofp- hydroxyl phenylpyruvic acid there ishomogenistic acid.
Due to this a disease alcaptonuria develops. In reality both C?f these
diseases are because of incomplete oxidation of phenyl pyruvic
acid. The reason for incomplete oxidation are mutated gene tt and aa.
3. Genes and Phenylketonuria:
Presence of phenylalanine causes a serious disease phenylketonuria.
The reason is that due to gene KK phenylalanine after oxidation is
unable to produce tyrosine. Actually due to mutation the gene KK is
converted into kk so that it is unable to produce the enzyme to convert
phenylalanine into tyrosine as shown in Fig-45.
Normal gene KK
Phenylalanine - - - - - - - - - - - - - - - -~ Tyrosine
Mutated gene kk
Phenylketonuria
Fig.45: Action of genes KK and kk in producing disease
phenylketonuria.
Gene and metabolism in neurospora:
Neurospora or red fungus is an important material for study of gene
action and biochemical studies. Beadle and Tayum and other scientists
based on their experiments have reported that for one biochemical
151
reaction only one gene is responsible. They have proved that enzymes
are produced by this fungus which controls several reaction activities.
These enzymes are controlled by special genes.
For example a normal neurospora has the gene V+ which produces
the enzyme trytophan synthetase which synthesize tryptophan . If
the gene V+ is mutated then it looses its capacity to producetrytophan
synthetase. So synthesis of tryptophan is not possible. But it has been
observed that if from normal neurospora fungus the obtained enzyme
trytophan synthetase extract is ·used for treatment of mutated fungus,
then the fungus will produce tryptophan. Therefore this experiment
shows the relationship between metabolism producing enzyme and
gene.
Gene and body size and body form:
Ductless glands in animals produces several types of harmones which
control characters such as body size and body form and many other
characters. Development of these ductless glands & other glands and
harmones are controlled by multiple gene complexes.
Gene and growth of plants:
In plants auxins or growth promoting hormones are found. Production
of growth promoting harmone is controlled by genes. For example in
maize the two varieties e.g. dwarf and normal are controlled by Na
and na. The scientist Vanoverbeck has found out that the gene na
produces an enzyme which disturbs the auxins, therefore growth of
the maize plant stops and the plant becomes dwarf.
152
(Part 11
Applied Genetics
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Chapter 1
Cancer Genetics
Cancer is a multistage carcinogenesis process during which normal
dividing cells accumulate multiple somatic mutations (point mu tations
and mutations due to chromosomal damage), and then become cancer
cells by several pathway events. Most human tumors are spontaneous
but some are age and sex related, and a few are inherited or familial.
There are many etiologicfactors such as genetic history, diet, lifestyle,
and environment may contribute to the development of these tumors.
In human latency period ranges from a few years to 30 years or more.
Cancer results from uncontrolled proliferation of monoclonalline with
aneuploidy. Almost all differentiated cells can become neoplastic by
the process called transformation. Malignant tumor (cancer) is a
multistage carcinogenic process (progression) requires more than
single genetic alteration (pathway events) to produce metastasis. Fig-
1 shows the sequence of Adenocarcinoma.
. .. ...........
HNPCC AoB........IJJ,l_
due $a tlft'P'l.one ftM'.Mion..
,"iUlA~,
Fig. 1: Sequence of Adenocarcinoma.
Immortalization refers to the process in which cells lose the capacity

to differentiate and then die, and thus continue to divide.
Immortalized cells, however, do not produce tumors and are non-
tumorigenic. Transformation refers to the process in which
immortalized cells acquire the capacity to produce tumors. Thus, the
transition from a normal cell to a cancer cell involves at least two
distinct steps: immortalization and transformation.
Accordingly, the process of immortalization in normal cells can be
called a "tumor initiation phase," whereas the process of
transformation of immortalized cells can be referred to as a "tumor
155
Gel.etics : Fundamentals and Applications
promotion phr'~'!." Thus, genes that cause immortalization can be

considered "tumor initiating genes," and genes that cause
immortalized cells to become cancer cells can be considered 11 tumor
promoting genes."
Neoplastic cells differ from normal cell in following respects:
They are immortal, loose contact inhibition, recognization and
communication, decreased adhesiveness, shows invasiveness and low
serum requirement, disorganized cytoskeletal structure, reduced
number of gap junctions, increased nuclear cytoplasmic ratio etc.
Uncontrolled growth of cells produce a cell mass called neoplasm or
tumor
The process of tumorgenesis:
Normal cell - -~ neoplasm (tumor) - - ~ Malignant tumor
(cancer)
Immortalization Transformation
Invasiveness
Metastasis
Following Fig- 2 describes the different stages of cancer
.~-~ IitIIl
--
-
~DNA
I
i
Fig. 2: Different stages of cancer.
156
Cancer Genetics
Cancer and cell cycle:

The uncontrolled proliferation of cancer is due to the failure of cell
cycle check point function at G/5 or G2/M or M phase. The check
points are the transitional periods where a decision is made to proceed
or halt progression through cell cycle. The decision is controlled by
physical interaction between CDKS & cyclins. Gl/5 check is controlled
by CDKS/ cyclins D & G2/M by CDKI/ cyclin B. Any mutation in genes
coding CDKS & cyclin may cause uncontrolled cell division leading
to malignancy.
Genetics of cancer:
Control of cell division is regulated in following ways.
(i) By genes: Genes suppressing cell division. Example tumor
suppressor genes.
(ii) Genes that normally function to promote cell division is proto-
oncogenes. Proto-oncogenes by gain of function mutation
becomes oncogene. There should be a balanced product of tumor
suppressor gene and proto-oncogene to maintain the normal cell
division.
The uncontrolled growth can be caused by following reasons :
(a) Loss of functional mutation of tumor suppressor gene.
(b) Permanent switch on of proto-oncogenes.
(c) Gain of functional mutation of proto-oncogene to oncogene.
These above three process lead to transformation, immortality. The
clonal expression of a malignant cancer may be due to following
reasons.
(a) Loss of function of differentiation genes.
(b) Epigenetic gene expression.
(c) Loss of heterozygosity of tumor suppressor genes.
(d) Non functional DNA repair mechanism.
Several hypotheses are there to explain the biology of carcinogenesis
and cancer those includes - two-stage model, chromosomal
aberrations, somatic mutation, activation of cellular oncogenes due
to point mutations and chromosomal re-arrangements, inactivation
of tumor suppressor genes due to point mutations or deletion of
chromosome segments and downregulation of" differentiation genes".
157
Two-Stage Model Hypothesis:

It says that at least two distinct stages of carcinogenesis, initiation
and promotion. Initiating events occurred at the nuclear DNA level,
whereas promoting events occurred at the epigenetic levels
(membranes, organelles, and post-translation protein modification).
This model is most valuable cancer model for public health to identify
many tumor- and antitumor-initiating agents, and treatment trials.
Chromosomal Damage Hypothesis:
This hypothesis is proposed by Boveri in 1914. According to this
hypothesis, chromosomal anomalies and damage by carcinogens
(carcinogens (ionizing radiation and chemicals) are the key initial
events in carcinogenesis. But chromosomal damage by itself is not
always sufficient to induce cancer in normal cells. However, this type
of damage can make the genome more sensitive to mutagens, and
thus can decrease the latency period during which downregulation
of a differentiation gene can occur as a random E7vent.
Somatic Mutation Hypothesis:
This hypothesis states that normal dividing cells become cancer cells
and exhibit immortalization or transformation after accumulation of
multiple, selective and specific somatic mutations due to genome
instability. Mutations that downregulates differentiation genes cause
immortalization of normal cells.
Viral Infection Hypothesis:
In rodents, some retroviruses (RNA viruses) like Rous sarcoma virus
(RSV), avian lymphomatosis viruses, murine leukemic viruses, feline
leukemic viruses and murine mammarytumor viruses causes cancer.
RNA viruses such as human-T-Ieukemia virus and hepatitis C virus
increases risk of T-cellleukemia and hepatic carcinoma respectively
in human. Genes from certain DNA viruses like large T-antigen (Lta)
gene from the SV40, E6, and E7 from HPV and E1A from the
adenovirus are responsible for immortalization of human cell.
Expression of the Lta protein of SV40 virus is for maintaining the cell
proliferation of immortalized rat fetal dopamine neurons.
Bacterial and Parasitic Infection Hypothesis:
Helicobacter pylori increase the risk of gastric cancer, Opistorchis viverrini
and Schistosoma haematobium are considered risk factors for
cholangiocarcinoma and urinary bladder cancer. A latency period of
158
Cancer Genetics
at least 14 years of chronic infection is needed for the development of

these tumors. Generation of free radicals and release of cytokines
during chronic infection may be one of the common intermediary
risk factors that increases the risk of cancer in infected organs.
Telomerase Hypothesis:
Telomerase enzyme has potential role in proliferation, aging, and
carcinogenesis. An increased activity of telomerase occurs subsequent
to immortalization and/ or transformation, and that it pays no direct
role in immortalization. Shortening of telomere length causes aging
and cell death.
Proto-Oncogene Hypothesis:
This hypothesis says that, proto-oncogenes are normal cellular genes
those when get mutated become oncogenous and cause cancer.
Classifications of Proto-Oncogenes:
According to the function of the particular proto-oncogene product
and function, Proto-Oncogenes can be classified in several groups.
Those are
1. Growth Factors:
The c-Sis gene (the v-sis gene is the oncogene in simian sarcoma virus)
encodes the PDGF B chain. The v-sis gene was the first oncogene to
be identified as having homology to a known cellular gene. The int-
2 gene encodes an FGF-related growth factor. The KGF (also called
Hst) gene also encodes an FGF-related growth factor and was
identified in gastric carcinoma and Kaposi's sarcoma cells.
2. Receptor Tyrosine Kinases:
The c-Fms ("fims") gene encodes the colony stimulating factor-l (CSF-
1) receptor and was first identified as a retroviraloncogene. The FIg
(" flag") gene (named because it has homology to the Fms gene, hence
fros-like gene) encodes a form of the FGF receptor. The Neu ("new")
gene was identified as an EGF receptor-related gene in an
ethylnitrosourea-induced neuroblastoma. The conversion of proto-
oncogenic to oncogenic Neu requires only a single amino acid change
in the transmembrane domain. The Trk ("track") genes encode the
NGF receptor-like proteins. The first Trk gene was found in a
pancreatic cancer. Subsequently, two additional Trk-related genes
were identified. These three are now identified as TrkA, TrkB and
159
TrkC. The Met gene encodes the hepatocyte growth factor(HGF}/

scatter factor (SF) receptor. The c-Kit gene encodes the mast cell
growth factor receptor.
3. Membrane Associated Non-Receptor Tyrosine Kinases:
The v-src gene was the first identified oncogene. The c-Src gene is
the archetypal protein tyrosine kinase. The Lck gene was isolated
from a T cell tumor line (LYSTRA cell kinase) and has been shown to
be associated with the CD4 and CD8 antigens of T cells.
4. G-Protein Coupled Receptors:
The Mas gene was identified in a mammary carcinoma and has been
shown to be the angiotensin receptor.
5. Membrane Associated G-Proteins:
There are three different homologs of the c-Ras gene, each of which
was identified in a different type of tumor cell. The Ras gene is one
of the most frequently disrupted genes in colorectal carcinomas.
6. Serine/fhreonine Kinases:
The Raf gene is involved in the signaling pathway of most RTKs. It is
likely responsible for threonine phosphorylation of MAP kinase
following receptor activation.
7. Nuclear DNA-Bindin&'I'ranscription Factors:
The Myc gene was originally identified in the avian myelocytomatosis
virus. A disrupted human c-Myc gene has been found to be involved
in numerous hematopoietic neoplasias. Disruption of c-Myc has been
shown to be the result of retroviral integration and transduction as
well as chromosomal rearrangements. The Fos gene was identified
in the feline osteosarcoma virus. The protein interacts with a second
proto-oncogenic protein, Jun to form a transcriptional regulatory
complex. The p53 gene was originally identified as a major nuclear
antigen in transformed cells. The p53 gene is the single most identified
mutant protein in human turnors. Mutant forms of the p53 protein
interfere with cell growth suppressor effects of wild-type p53
indicating that the p53 gene product is actually a tumor suppressor.
Oncogene Hypothesis:
Gain of function mutation or overexpression of cellular proto-
oncogenes is the primary events that initiate neoplastic changes. Viral
160
Cancer Genetics
oncogenes such as v-src, v-ras, and v-myc were isolated from

retroviruses and were found to be tumorigenic and are counterparts
(cellular proto-oncogenes)from the mammalian genome. But mutation
in cellular proto-oncogenes cannot be considered an initial event in
human carcinogenesis but are the secondary events in human
carcinogenesis. Over expression of multiple oncogenes such as ras,
myc, c-erbB2, and Bcl2 occur in human lung cancer and 30% of
adenocarcinomas show mutation in K-ras, and overexpression of c-
erbB2 is found in 25 % of non-small-celllung cancer (NSCLC).
List of oncogene and their related cancers
Oncogene Function Cancer
Abl Tyrosine kinase activity Chronic myelogenous leukemia
Chronic myelogenous and acute
abl/bcr New protein created by fusion
!ym~hoticleukemia
Encodes a protein-
akt-2 Ovarian cancer
serine/ threonine kinase
Encodes a receptor tyrosine
Alk Lymphomas
kinase
Amll/mtg8 New protein created by fusion Acute leukemias
Encodes a receptor tyrosine
Axl Hematopoietic cancers
kinase
Block apoptosis (programmed
bcl-2, 3, 6 B-celllymphomas and leukemias
cell death)
c-myc Cell proliferation and DNA Leukemia; breast, stomach, lung,
synthesis cervical, and colon carcinomas;
neuroblastomas and
Zlioblastomas
Egfr Tyrosine kinase Squamous cell carciI:toma
erg/c16 New protein created by fusion Myeloid leukemia
Glioblastomas, and squamous cell
erbB Tyrosine kinase
carcinomas
erbB-2
Breast, salivary gland and
(originally Tyrosine kinase
ovarian carcinomas
neu)
Fos Transcription factor for API Osteosarcoma
Fps Tyrosine kinase Sarcoma
Gli Transcription factor Glioblastomas
Membrane associated G
Gsp Thyroid carcinoma
Iprotein
New protein created by gene
HER2/neu Breast and cervical carcinomas
fusion
Over-expression of DNA
Hoxll Acute T-cellleukemia
binding protein
161
Hst Encodes fibrobIast growth factor Breast and squamous cell

carcinomas
IL-3 Over expression of protein Acute pre B-cell leukemia
int-2 Encodes a fibroblast growth factor Breast and squamous cell
carcinomas
Jun Transcription factor for API Sarcoma
K-sam Encodes growth factor receptors Stomach carcinomas
Cell proliferation and DNA
L-myc Lung carcinomas
synthesis
Mdm-2 Encodes a pS3 inhibitor Sarcomas
Encodes p16 a negative growth
MLM Melanomas
regulator that arrests the cell cycle
Hereditary nonpolyposis
MSH2 Mismatch repair in DNA
colorectal cancer
Mtg8/aml
New protein created by fusion Acute leukemias
1
Encodes a transcription factor with Colon carcinoma and
Myb
DNA binding domain leukemias
Neu Tyrosine kinase Glioblastomas and squamous
(nowerb- cell carcinomas
2)
N-myc Cell proliferation and DNA Neuroblastomas,
synthesis retinoblastomas and lung
carcinomas
NPM/ALK New protein created by fusion Large cell lymphomas
nrg/rel New protein created by fusion B-celllymphoma
Guanine nucleotide exchange
Ost Osteosarcomas
factor
Relocation of transcription factor Lympho-plasmacytoid B-cell
pax-S
to the IgH gene lymphoma
Encodes cyclin Dl that is Breast and squamous cell
PRAD-l
imp_ortant in Gl of the cell cycle carcinomas
Raf Serine/ threonine kinase Many cancer types
Involved in signal transduction of Lung, ovarian, and bladder
rasK
the cell carcinoma
Involved in signal transduction of
rasN Breast carcinoma
the cell
Ros Tyrosine kinase Sarcoma
Src Tyrosine kinase Sarcomas
Over-expression of transcription
tal-l,2 Acute T-cell leukemia
factor
tan-l Over-expression of2rotein Acute T-cell leukemia
Guanine nucleotide exchange
Tiam-l T-Iymphoma
factor
TSC2 GTPase activator Renal and brain tumors
Trk Recombinant fusion I'rotein Colon and thyroid carcinomas
162
Cancer Genetics
Tumor Suppressor Gene (Anti-Oncogene) Hypothesis:

This concept was derived from the cytological study of retinal cells
and retinoblastoma cells in which one of the alleles of chromosome
13 was missing from the retinal cells and both were missing from the
retinoblastoma cells. This suggested that chromosome 13 must carry
a tumor suppressor gene, the loss of which was essential for tumor
formation. Wild-type Rb gene also inhibited the growth of normal
cells.
Another well-known tumor suppressor gene, p53 and mutations in
p53 are present in up to 50% of NSCLC and in 80% of SCLC.
List of tumor suppressor genes and their related cancers
Syndrome! Cloned Gene Function Chromosomal TumorTypes
Cancer Location
Li-Fraumeni P53 tumor cell cycle l7pl3 brain tumors,
Syndrome suppressor regulation, sarcomas,
apoptosis leukemia, breast
cancer
Familial RBl cell cycle l3ql4 retinoblastoma,
Retinoblasto tumor suppressor regulation osteogenic
ma sarcoma
Wilms WTl transcrip- llpl3 pediatric kidney

Tumor tumor suppressor tional cancer
regulation
Neurofibro NFl catalysis of l7qll.2 neurofibromas,
matosis protein=neurofib RAS sarcomas,
Typel rominl inactivation gliomas
tumorsuppressor
Neurofibro NF2 linkage of cell 22ql2.2 Schwann cell
matosis protein = merlin membrane to tumors,
Type 2 or cytoskeleton astrocytomas,
neurofibromin 2 meningiomas,
tumorsuppressor ependynomas
Familial APC signaling 5q2l colon cancer
Adenomato tumor suppressor through
us Polyposis adhesion
molecules to
nucleus
Tuberous TSCl 9q34 facial
sclerosis 1 protein = hamarti angiofibromas
n
tumorsuppressor
163
Tuberous TSC2 GIPase 16 benign growths

sclerosis 2 protein = tuberin activation (hamartomas) in
turnor suppressor many tissues,
astrocytomas,
rhabdomyosarco
mas
Deleted in DPC4 regulation of 18q21.1 pancreatic
Pancreatic also known as TGF-BMP carcinoma, colon
Carcinoma 4 Smad4 signal cancer
tumorsuppressor transduction
Deleted in DCC transmembra 18q21.3 colorectalcancer
Colorectal turnor suppressor ne receptor
Carcinoma involved in
axonal
guidance via
netrins
Familial BRCA1 repair of 17q21 breast and
Breast turnor suppressor double ovarian cancer
Cancer strand breaks
by
association
with Rad51
iprotein
Familial BRCA2 Similar to 13q12.3 breast and
Breast turnor suppressor BRCAl? ovarian cancer
Cancer
Peutz- STK11 potential 19p13.3 hyperpigmentati

Jeghers tumor suppressor regulation of on, multiple
Syndrome protein = serine- vascular hamartomatous
threonine kinase endothelial polyps,
11 growth colorectal, breast
factor and ovarian
(VEGF) cancers
Ipathway
Hereditary MSH2 DNA 2p22-p21 colorectal cancer
Nonpolyposi turnor suppressor mismatch
s Colorectal repair
Cancer type
1
HNPCCl
164
Cancer Genetics
Hereditary MUll DNA 3p2l.3 colorectalcancer

Nonpolyposi tumor suppressor mismatch
s Colorectal repair
Callcer type
2
HNPCC2
vonHippel- VHL regulation of 3p26-p25 renal cancers,

Lindau tumor suppressor transcription hemangioblasto
Syndrome elongation mas,
pheochromocyto
ma
Familial CDKN2A inhibits cell- 9p2l melanoma,
Melanoma protein =cyclin- cycle kinases pancreatic
dependent kinase CDK4and cancer, others
inhibitor 2A CDK6
tumor suppressor
Gorlin PfCH transmembra 9q22.3 basal cell skin
Syndrome: protein = patched nereceptor cancer
Nevoid tumor suppressor for hedgehog
basal cell signaling
carcinoma protein
syndrome
(NBCCS)
Multiple MENl Unknown llq13 parathyroid and

Endocrine tumor suppressor pituitary
Neoplasia adenomas, islet
Type 1 cell tumors,
carcinoid
Multiple RET,MEN2 transmembra lOqll.2 medullary
Endocrine nereceptor thyroid cancer,
Neoplasia tyrosine type2A
Type 2 kinase for pheochromocyto
glial-derived ma,mucosal
neurotrophic hartoma
factor
i(GDNF)
Beckwith- p57,I<IP2 cell cycle llp15.5 Wilms tumor,
Wiedmann regulator adrenocortical
Syndrome
. cancer,
hepatoblastoma
165
Hereditary MET transmembra 7q31 renal papillary

papillary ne receptor cancer
renal cancer for
(HPRq hepatocyte
growth
factor (HGF)
Hereditary HPCl and PRCAl RNaseL lq24-q25 prostate .cancer
prostate are same involved in
cancer designation mRNA
numerous ribonuclease L degradation
loci: (RNaseL) maps to
HPCl(pRCA this locus
l),HPCX,
MXIl,KAIl,
PCAP
Ataxia ATM gene product llq22.3 lymphoma,

telangiectasi 4 likely halts cerebellar ataxia,
a (AT) complementation cell cycle immunodeficien
groups: ATA, after DNA cy
ATC, ATD, ATE, damage
are associated
with mutations in
theATMgene
Bloom BLM DNA 15q26.1 solid tumors,
syndrome helicase immunodeficien
RecQ cy
protein-like-3
Xeroderma XPA-XPG DNA repair XPA=9q22.3 skin cancer
pigmentosu helicases, XPC = 3p25
m(XP) nucleotide XPD=19q13.2
7 excision -q13.3
complentatio repair XPE=llp12-
ngroups pll
XPF=16p13.3-
XPAXPCXP p13.13
D
Function of p53:
TP53 level in Gl of a healthy cell is very low. But damage in DNA
rapidly increases the level. Rising of TP53 activates p21 gene product
and cell cycle is arrested at GljS point providing the cell DNA repair
time before initiation of DNA replication in 5 phase.
When both the copies of p53 are mutated, it no longer can produce
p21 inhibitor to prevent the entry to 5 phase. It results mutated DNA
166
Cancer Genetics
replication that leads production of transformed cells. Following Fig-

3 describes different pathway events involved in carcinogenesis
Fig. 3: Pathway events in cancer showing involvement of different types

of oncogenes and P53.
Differentiation Gene Hypothesis:
A gene, which induces terminal differentiation in normal cells referred
to as a differentiation gene and its mutation may be an initiating event
that causes cell to become immortalized and that mutations (point
mutation or overexpression) in cellular oncogenes, growth regulatory
genes, and tumor suppressor genes may represent secondary events
that convert immortalized cells into transformed cells. Mutation in
differentiation gene prevents the cell from undergoing terminal
differentiation and subsequent cell death and consequently, the cells
continue to proliferate (hyperplasia), leading to the formation of pre-
neoplastic lesions such as adenomas. Differentiation gene is regulated
by at least two sets of genes, "differentiation suppressor genes" and
tumor suppressor genes, in an opposite manner. For example,
inactivation of a "differentiation suppressor gene" or activation of a
tumor suppressor gene downregulates the expression of a
differentiation gene. Mutation in a differentiation gene is sufficient
to downregulate this gene. The evidence for the existence of a
differentiationsuppressor gene is indirect. A differentiation gene may
be downregulated by one of the following mechanisms: mutation
within the differentiation gene; overexpression of "differentiation
suppressor genes;" and inactivation of tumor suppressor genes (Fig-
4). Some candidate differentiation genes (c-fos, c-fes, and RAG-l gene
activator) that are upregulated during cAMP-induced terminal
differentiation of NB cells.
167
Cardnoaens
(physical. chemical and viral)
Mutations in Dividill8 Cella
I~
int mullllions andIor chn>mosnmal <lama
No C.IIDi~sion
Conscqucnccon
or Differentiation
!-.#,
-€>
Genes Not Involved

in Division
or Dift'enmlialion
I
No Consequence on
.Cell Division
or Diffcrenliation
Fig. 4: Differentiation Gene hypothesis.

Cancer: Types, Diagnosis and treatment:
Cancer is an abnormal, continuous and uncontrolled multiplication
of cell that may grow into adjacent tissue or spread to distant parts
of the body (metastasis). The mass of cancer cells eventually become
large enough to produce lumps, masses or tumors that can be
detected, which can be benign or malignant:
Benign tumors: These are not cancerous and can usually be removed
and do not spread to other parts of the body, and the cells do not
invade other tissues
Malignant or metastatic tumors (cancer): These are cancerous and
can invade and damage nearby tissues and organs and metastasize.
This type of cancer can spread from its original location to other parts
of the body by direct extension or invasion into adjacent tissues or
, by Systemic spread throughout the body by blood and lymphatic
system or by cerebrospinal fluid (Fig-5)
168
---------- ---- ---
Cancer Genetics
Fig. 5: Process of metastasis.

General categories of cancers:
There are four general categories of cancer and adenocarcinomas are
the most common type.
I. Carcinomas are cancers that occur in epithelial surfaces - the cells
that form the outer surface of the body to line or cover the body's
cavities, tubes and passageways.
11. Adenocarcinomas are cancers that form on a glandular surface,
such as the lung, breast, prostate, ovary, or kidney.
Ill. Sarcomas are cancers that occur in supporting structures, such
as bone, muscle, cartilage, fat or fibrous tissue.
IV. Leukemias and lymphomas are cancers that occur in blood cell
elements.
Carcinoma and, sarcoma fall under solid tissue tumor. Lymphoma
and leukemia are soft tissue cancer.
Tissue and organ specific cancers such as liver cancers are those that
originated from a liver cell are similarly Brain cancers, nerve cancers,
melanomas, and certain testicular and ovarian cancers etc.
169
Diagnostic procedures for cancer:

When the symptoms of cancer appears, the diagnosis can be done by
several methods. First a detailed medical history (family and personal)
is to be considered to know the hereditary relation, then physical
examination can be done. Other diagnostic procedures that may be
requested include
Imaging tests:
1) X-ray, computed tomography (CT or CAT scan) for brain or
other internal organs, lymph nodes cancers.
2) Radionuclide scan - to detect any abnormal areas or tumors.
3) Ultrasound - for abdominal organ tumors, such as the uterus,
liver, and kidneys.
4) Magnetic resonance imaging (MRI) for internal organ or
structure, especially the brain and spinal cord, nodules in bones
or lymph nodes.
5) Endoscopy - allow to see inside the hollow organs, such as the
bladder or uterus. Biopsy samples are taken through this.
Laboratory tests:
To examine blood, urine, other fluids or tumor tissue by microscopy
and by immunohistochemistry.
Biopsy:
It is an invasive method to remove a sample of the suspicious tissue
for examination in a laboratory.
After diagnosis stage of progression can be evaluated to determine
the extent (spread) of the cancer which can be done by staging. Staging
is important for treatment, prognosis and treatment results. Stage
refers to the extent, or the size, of the cancer. Each cancer, by organ,
has its own staging system.
Stages of cancer:
There are five stages in cancer and are non reversible
1. Stage 0 or carcinoma in situ: It is very early cancer. The abnormal
cells are found only in the first layer of cells of the primary site
and do not invade the deeper tissues.
2. Stage I: Cancer involves the primary site, but has not spread to
nearby tissues.
170
Cancer Genetics
a. Stage lA: a very small amount of cancer - visible under a

microscope - is found deeper in the tissues.
b. Stage IB: a larger amount of cancer is found in the tissues.
3. Stage 11: Cancer has spread to nearby areas but is still inside the
primary site.
a. Stage lIA: cancer has spread beyond the primary site.
b. Stage lIB: cancer has spread to other tissue around the
primary site.
4. Stage Ill: Cancer has spread throughout the nearby area.
S. Stage IV: Cancer has spread to other parts of the body.
a. stage IVA: cancer has spread to organs close to the pelvic
area.
b. stage !VB: cancer has spread to distant organs, such as the
lungs.
Recurrent cancers again come back (recurred) after it has been treated
but not cured properly.
Treatment of cancer:
The goal of cancer treatment is successful treatment, prevention,
prolonged life and palliation (pain relief).
Specific treatment for a cancer is determined by several factors like-
overall health and medical history, extent of the disease - type, grade,
stage and location, tolerance for specific medications, procedures, or
therapies, expectations for the course of the disease etc.
The conventional and primary methods of cancer treatment are:
1. Surgery
2. Radiation therapy
3. Chemotherapy
4. Hormone therapy
5. Immunotherapy / biological therapy
6. Adjuvant therapy - combining two or more treatments
7. Prophylactic or preventive treatment.
S. Gene Therapy
171
Genetics : Futldamentals and Applications
Surgery:
It is a surgical procedures used to diagnose or destroy cancerous
tissue include by several surgical techniques like biopsy, endoscopy,
laparoscopy, laparotomy, laser surgery, cryosurgery, electrosurgery,
excisional etc.
Radiation Therapy:
It uses special kinds of energy waves or particles to fight cancer
(destroy cancer cells or prevent cells from growing or reproducing).
Depending on the type and location of the cancer, the therapy varies.
Radiation Therapy provides a cure for cancer, control the disease, or
help relieve its symptoms. Radiation treatments are painless and
usually last a few minutes.
Procedures in Radiation Therapy:
Simulation process: It is "mapping" out the position for treatment
and the exact location of body (referred to as treatment field or port)
wh~re the radiation will be given (the simulation process). Imaging
studies may are performed duringthe simulation process to plan how
to direct the radiation during the treatments.
Treatment plan: After simulation process treatment plan is
determined, including the type of machine to use, the amount of
radiation that is needed and the number of treatments that will be
given.
Types of radiation therapies:
Depending on the type of cancer, method of Radiation therapy varies.
The main considerations are location of the cancer and the patient's
health. Radiation therapy can be used in combination with other
treatments. The most common types of radiation therapy are -
External radiation (external beam therapy): Here radiation is
administered by a large machine that points the energy waves directly
at the tumor to kill cancer cells. Special shields are made to protect
the tissue surrounding the treatment area.
Internal radiation (brachytherapy, implant radiation): Here a high
dose of radiation is given inside the body as close to the cancer as
possible by administering a higher dose of radiation in a shorter time
span when compared with external radiation. The radiation treatment
may be swallowed, injected or implanted directly into the tumor.
Some of the radioactive implants are called "seeds" or "capsules".
172
Cancer Genetics
Chemotherapy:
Chemotherapy is the use of anticancer drugs to treat cancerous cells.
It is one of the most common treatments for cancer. Different groups
of drugs work in different ways to fight cancer cells by interfering
with the cancer cell's ability to grow or reproduce. But there are
many side effects of chemotherapy.
Chemotherapy can be given as a pill, as an intramuscular injection,
intravenously (directly to the bloodstream; also called IV), topically
(applied to the skin) or directly into a body cavity
Some of the chemotherapy drugs and their side effects
Chemotherapy Drug Possible Side Effects (Not all side effects
are listed. Some of those listed may be
short-term side effects; others are long-
term side effects.)
Carboplatin (paraplatin): Usually Decrease in blood cell counts, hair loss
given intravenously (IV) and used (reversible)
for cancers of the ovary, head and Confusion, nausea, vomiting and/or
neck, and lung. diarrhea (usually a short-term side effect
occurring the first 24 to 72 hours following
treatment)
Cisplatin (platinol, Platinol-AQ) Decrease in blood cell counts, allergic
Usually given intravenously (IV) reaction, including a rash and/ or labored
and used for cancers of the breathing, nausea and vomiting that
bladder, ovary and testicles. usually occurs for 24 hours or longer
ringing in ears and hearing loss,
fluctuations in blood electrolytes, kidney
dama~e.
Cydophosphamide (Cytoxan, Decrease in blood cell counts, nausea,
Neosar): vomiting, abdominal pain, decreased
Can be given intravenously (IV) or appetite, hair loss (reversible), bladder
orally and used for lymphoma, damage, fertility impairment, lung or heart
breast cancer, and ovarian damage (with high doses), secondary
carcinoma mali~cies (rare).
Docetaxel (faxotere): Usually Decrease in blood cell counts, nausea,
given intravenously (IV) and used vomiting, abdominal pain, diarrhea,
for breast cancer, lung and decreased appetite, hair thinning, rash,
prostate numbness and tin~lin~ in hands and feet.
Doxorubicin (Adriamycin): Decrease in blood cell counts, mouth ulcers,
Usually given intravenously (IV) hair loss (reversible), nausea and vomiting,
and used for breast cancer, heart damage
lymphoma, and multiple myeloma
Erlotinib (farceva) Usually given Ras~ and other skin changes and diarrhea
orally and
used for non small celllun~ cancer
173
Etoposide (VePesid): Can be Decrease in blood cell counts, hair loss

given intravenously (IV) or orally (reversible)
and used for cancers of the lung, nausea and vomiting, allergic reaction
testicles, leukemia and lymphoma mouth ulcers
low blood pressure (during
administration), decreased appetite,
diarrhea and abdominal pain,
bronchospasm and flu-like symptoms
Fluorouracil (5-FU): Usually given Decrease in blood cell counts, diarrhea,
intravenously (IV) and used for mouth ulcers
cancers of the colon, breast, Photosensitivity, dry skin
stomach, and head and neck
Gemcitabine (Gemzar): Usually Decrease in blood cell counts, nausea and
given intravenously (IV) and used vomiting, fever and flu-like symptoms and
for cancers of the pancreas, breast, rash
ovary and lung
Imatinib mesylate (Gleevec): Nausea and vomiting, fluid retention
Usually given orally and used for (swelling around ankles, eyes), muscle
chronic myelogenous leukemia cramps, diarrhea , gastrointestinal bleeding
(CML) and gastrointestinal stromal and rash
tumor (GIST)
lrinotecan (Camptosar): Usually Decrease in blood cell counts, diarrhea and
given intravenously (IV) and used hair loss (reverSible)
for cancers of the colon and rectum
Methotrexate (Folex, Mexate, Decrease in blood cell counts, nausea and
Amethopterin): May be given vomiting
intravenously (IV), intrathecally mouth ulcers, skin rashes and
(into the spinal column), or orally photosensitivity
and used for cancers of the breast, dizziness, headache or drowsiness, kidney
lung, blood, bone, and lymph damage (with a high-dose therapy), liver
system damage, hair loss (reversible) and seizures
Paclitaxel (Taxol, Abraxane): Decrease in blood cell counts, allergic
Usually given intravenously (IV) reaction, nausea and vomiting, loss of
and used with cancers of the appetite, change in taste, thin or brittle hair
breast, ovary, and lung joint pain (short term), numbness or
tingling in the fingers or toes
Sorafinib (Nexavar): Usually high blood pressure (during first few weeks
given orally of treatment), rash, other skin changes,
for advanced kidney cancer diarrhea, fatigue, hair loss, nausea and
vomiting
Sunitinib (Sutent): usually given Diarrhea, nausea and vomiting, mouth
orally ulcers, upset stomach skin changes,
for gastrointestinal stromal tumor including skin discoloration and rash,
(GIST) and kidney cancer fatigue, high blood pressure, bleeding,
swelling and taste disturbance
174
Cancer Genetics
Topotecan (Hycamtin): Usually Decrease in blood cell counts, diarrhea,hair

given intravenously (IV) for loss (reversible), nausea and vomiting
cancers of the ovary and lung
Vincristine (Oncovin, Vincasar Numbness or tingling in the fingers or toes,
PFS): Usually given intravenously weakness, loss of reflexes, jaw pain, hair
(IV) for leukemia and lymphoma loss (reversible), constipation or abdominal
cramping
Vinblastine (Velban): Usually Decrease in blood cell counts, hair loss
given intravenously (IV) for (reversible), constipation or abdominal
lymphoma and cancers of the testis cramping, jaw pain, numbness or tingling
and head and neck in the fin~ers or toes
Hormone Therapy:
Hormones help some types of cancer cells to grow, such as breast
cancer and prostate cancer and in other cases; hormones can kill cancer
cells, slower cancer cells growth or stop their growth. Hormone
therapy for cancer treatment involves surgically removal of a hormone
producing gland that helps growth of cancer cells. Hormone therapy
depends upon many factors such as the type and size of the tumor,
the age of the person, the presence of hormone receptors on the tumor
etc.
H the hormone receptor test indicates that the hormones are affecting
cancer, the cancer may be treated in one of following ways:
1. treating cancer cells to keep them from receiving the hormones
they need to grow
2. treating the glands that produce hormones to keep them from
making hormones
3. surgery to remove glands that produce the hormones, such as
the ovaries that produce estrogen, or the testicles that produce
testosterone
If hormone therapy is given before the primary treatment, it is called
neoadjuvant treatment. Neoadjuvant treatments help to kill cancer
cells and contribute to the effectiveness of the primary therapy. If
hormone therapy is given after the primary cancer treatment, it is
called adjuvant treatment. Adjuvant therapy is given to improve the
chance of a cure.
Biological Therapy:
Also called immunotherapy, biological response modifier therapy,
or biotherapy. It uses the body's immune system to fight cancer.
Biological therapies are designed to boost the immune system, either
175
directly or indirectly, by assisting in the following:

1. Making cancer cells more recognizable by the immune system,
and therefore more susceptible to destruction by the immune
system.
2. Boosting the killing power of immune system cells.
3. Changing the way cancer cells grow, so that they act more like
healthy cells.
4. Stopping the process that changes a normal cell into a cancerous
cell.
5. Enhandng the body's ability to repair or replace normal cells
damaged or destroyed by other forms of cancer treatment, such
as chemotherapy or radiation.
6. Preventing cancer cells from spreading to other parts of the body
There are many different types of biological therapies used in cancer
treatment.
Biological response modifiers (BRMs) include nonspecific
immunomodulating agents, interferons, interleukins, colony-
stimulating factors, monoclonal antibodies, cytokine therapy, and
vaccines those can boost the body's ability to fight the disease, direct
the immune system's disease fighting powers to disease cells and
strengthen a weakened immune system.
Nonspecific immunomodulating agents:
Stimulate the immune system to produce more cytokines and
antibodies to fight cancer.
Interferons (IFN): Naturally occurs in the· body and also can be
produced in the laboratory for use in cancer therapy. IFN improves
immune system and acts against cancer cells by slow down growth
or by changing cancer cell to normal behaviour. Some interferons
may also stimulate natural killer cells (NK) cells, T cells, and
macrophages that help to fight cancer cells.
Interleu,ldns (IL): Interleukins are proteins (cytokines) and can
stimulate the growth and activity of immune cells, such as lymphocytes,
which work to destroy cancer cells.
Colony-stimulating factors (CSFs): Can encourage stem cells within
the bone marrow to produce more blood cells. CSFs are given, along
with chemotherapy, to boost the immune system.
Monoclonal antibodies: Are produced in the laboratory. They can
176
Cancer G~tics
bind to cancer cells and when cancer-destroying agents are introduced

into the body, they seek out the antibodies and kill the cancer cells.
For example Trastuzumab (Herceptin®) is used for breast cancer and
Rituximab (Rituxan®) for lymphoma.
Cytokine therapy: Cytokine helps immune system to recognize and
destroy cancerous cells. Cytokine therapy kills cancer cells and
prevents tumors from growing.
There are few side effects of biological therapy, which often mimic
flu-like symptoms and varies according to the type of therapy. These
are fever, chills, nausea, vomiting, loss of appetite, fatigue etc.
Specifically, cytokine therapy often causes fever, chills, aches and
fatigue. Other side effects include a rash or swelling at the injection
site. Therapy can cause fatigue and bone pain and may affect blood
press~e and the heart.
177
Chapter 2
Apoptosis
Apoptosis is the term coined by Andrew Wyllie in 1972 to describe
morphological description of a dying cell that contrast with necrosis.
Programmed cell death (PCD) is a term used to describe cells that die
at predictable time and place during development. Programmed cell
death is a gene directed cellular suicide mechanism that eliminates
unwanted, superfluous cells during development as well as tissue
homeostasis, control proliferation and differentiation and defends
against viral infection. All Programmed cell deaths are apoptotic and
so these terms are used interchangeably.
Programmed cell death is associated with ALPS- Autoimmunity,
Myoicordial infraction, Stroke, Diabetes, Neurodegenerative diseases,
Alzheimer's disease, Infertility, Sepsis, Viral infection etc.
When Apoptosis does occur:
In embryonic • Tissue developmental programs, which control
and fetal sculpturing of embryonic form.
development • Limb and organ differentiation and
development.
• Developmental organization of nervous system.
• Elimination of self-reactive components of
immune system.
In adult • On stimulation by T-Lymphosites.

• In response to DNA damage, viral infeCtion or
transformation, radiation etc.
• In certain organ or tissue on withdrawal of
supporting hormone.
Characteristics of Apoptosis:
1. Cytoplasm shrinks with out membrane rupture.
2. Blabbing of plasma membrane and nuclear membrane.
3. Chromatin condenses and migrates to nuclear membrane.
178
Apoptosis
4. Inter-chromosomal cleavage in nucleosomal size by Mg+2

dependent endonuclease leads to laddering of DNA at
nucleosomal repeat length of 200bp.
5. Fragmentation of cell with retention of cell membrane.
6. Flipping up phosphatidal serine from outer plasma membrane.
7. Cell contents are packed in membrane bounded bodies (apoptotic
bodies).
8. Internal organneles still remain functional, mitocondria swells
and engulfed by neighbors.
9. Epitopes appear on plasma membrane, making cell as
phagocytotic target.
10. No spillage and inflammation.
Differences between apoptosis and necrosis
Apoptosis Necrosis
1. A normal physiological response 1. Pathological response to
to specific suicide signals or lack cellular injury.
of survival signals.
2. Chromatin condenses. 2. Nuclear swelling and
Cell shrinks. chromatin clumps.
3. Preservation of cell organelle. 3. Cell swells
4. Blabbing of plasma membrane 4. Mitochondria swell and
and nuclear membrane. rupture.
5. No lysis takes place. 5. Plasma membrane lyses
6. Rapid engulfment of dying cells 6. Slow phagocytosis
by neighboring cells.
7. Prevent inflammation and pus 7. Triggers inflammation and
formation. pus formation
8. DNA breaks early in 8. Cell ruptures and out of cell
internucleosomal pattern. content.
9. DNA breakage by gene 9. DNA breaks later and
activated, endonucleases randomly, DNA breakage by
random injury and ATP
depleted
10. Moderate Calcium influx. 10. Massive Calcium influx.
179
SmallbIebs
form.
The bIebs fU80

and become
targer.no
organ......
fOcaltd In the
bIeba.
.....
The eel bnNIII:s
apoptotic
bOcI8t;lIWl
OfgInIIIIIare
. . ftmcIionaI.
Fig. 6: Differences between apoptosis and necrosis.
Genes involve in apoptosis:

Gene Family Mammals C. elegans Drosophila
Caspase (1-14) Casp-l to Casp-14 Ced-3 (casp-9) Ocp-l, drICE

Bcl-2 (19)
Survival factors Bcl-2, Bel-Xl Ced-9
Death promoter Bax, Bad, Bak, Bid, Drob-l, buffy

Bim, Bik
Death domain proteins Fas, CD-95, APO- Ced-3, Csp-l, dredd (Casp-8),
( these are apoptosis 1, Csp-2 drone, dream.
inducer and eontain long TNFR?
lI>rodomain)
lAP N-AIP, ClAP-I, Ces,Mes dlAP-I, dIAP-2,
(Inhibitor Apoptotie X-lAP, CIAP-2, Deterin.
Proteins) DIAP-I,
SURVIVIN
180
Apoptosis
AIF
( "nuclear Apoptotic AIF ? dAIF
Inducing Factors,
released from
Mitochondria}
Ced-4 APAF-I Ced-4 Ark (apaf-I
(Apoptosis activator) (apoptotic related killer)
protease activating
factor-I)
Acinus Acinus ? Acinus-d
Mechanism of apoptosis:
There are 3 different mechanisms of apoptosis (Fig-7)
A. Death signals generated within the cell (Intrinsic/ mitochondrial
pathway).
B. Activation of death activators by extrinsic death signals (Extrinsic
or death receptor signaling pathway).
C. By reactive oxygen species and DNA damage.
Intrinsic or mitochondrial pathway:
Mitochondrial outer membrane displays Bcl-2 on their surface in
healthy cells due to internal damage to the cell (e.g., from reactive
oxygen species, DNA damage, UV radiation) causes BcI-2 to activate
related protein, Bax, that causes leackage in the outer mitochondrial
membrane to release cytochrome-c from mitochondria. Upon released
cytochrome-c binds to Apaf-l with the help of ATP and form aggregate
called apoptosomes. Apoptosome then binds to and activates caspase-
9. Caspase-9 cleaves and do activates other down stream caspases or
executioner caspases (caspase-3 and -7) leading to digestion of
structural proteins in the cytoplasm, chromosomal DNA degradation
• and phagocytosis of the cells those are the phenomena of apoptosis.
Extrinsic or death receptor signaling pathway:
Fas and the TNF receptor are integral membrane proteins. Their
receptor domains remain exposed at the cell surface. When
complementary death activator (FasL and TNF respectively) binds
to these receptors, it transmits a signal to the cytoplasm leading to
caspase 8 activation. Activated Caspase 8 initiates a cascade of Caspase
activation (caspase-3, 6, and 7) that causes apoptosis.
181
Apoptosis-Inducing Factor (AIF):

Neuronsdoes not use Caspases for self-destructuction. It uses
.
Apoptosis-inducing factor (AIF) which is normally located in the
mitochondrial intermembrane space. Upon receiving death stimulus,
AIF is released out from the mitochondria and migrates into the
nucleus, where it binds to DNA and cleaves it causing cell death.
Free Radical signaling pathway:
Oxidative stress can trigger apoptosis. Activated neutrophils produce
reactive oxygen species like superoxide free radicals those can induce
endothelial cell apoptosis. Endothelial cells also produce reactive
oxygen species inside the cell during reperfusion injury following
ischemia. Superoxide dismutase (SOD) converts highly reactive and
damaging superoxide free radicals to peroxides. Peroxides and
hydroxyl radicals activate NF-kB and activate expression of
inflammatory genes including adhesion molecules, TNF and IL-S.
---.-
---..,-,-
---f _ __
---1-"'--
-----.-
---i-_-
- . -..
~T __
~T __
.........
~=
~- .. - ~
Fig. 7: General Apoptotic pathways.
182
Apoptosis
Other pathways are:

• Apoptosis Pathway mediated by Granzyrne A.
• HIV Induced T Cell Apoptosis.
• Induction of apoptosis through DR3 and DR4/5 Death Receptors.
• Multiple antiapoptotic pathways from IGF-IR signaling lead to
BAD phosphorylation.
• TSP-l Induced Apoptosis in Microvascular Endothelial Cell.
Fig-8: Apoptosis in C. elegans.
Fig.9: Apoptosis in Drosophila: Intrinsic and extrinsic pathways.

Cancer and apoptosis
Cancer is a multistage carcinogenic process where there is a net
accumulation of atypical cells arising from excess proliferation, an
insufficient apoptosis or a combination of the two. There are also
183
various side effects of chemotherapy. Thus from therapeutic point of

view, the best strategy is 11 induced apoptosis in the neoplastic cell
line without affecting the normal cells of the body. Several diatary
phytochemicals are nowadays are used for chemoprevention those
do not cause side effects. The list provides few of those
phytochemicals used for cancer prevention.
A list of antioxidant, anticancer and dietary phyto-chemicals those
can induce apoptosis to prevent various types of cancers:
Phytochemical Source Action
Geraniol Bergamot, carrot, coriander, Inhibits Colon and Pancreatic
lavender, lemon, lime, cancer Growth
nutmeg, orange, blueberry
and blackberry
Cyanidin present in skin of most red Exhibits an inhibitory effect on
coloured berries such as the migration and invasion of
bilberry, blackberry, cherry, human lung cancer celIlines.
cranberry, elderberry, and
also in apples, pears, peaches
and plums
Allicin Predominant thiosulfinate in Lowers the risk of stomach
garlic (Allium sativum) and cancer.
responsible for the typical
and offensive odor of garlic.
lndole-3- Found in cruciferous Reduces the risk of breast,
Carbinol vegetables such as cabbage, prostate and cervical cancer
cauliflower, broccoli, kale and
brussels sprouts.
Isothiocyanates Found in cruciferous Helps to prevent lung cancer
vegetables such as broccoli, and esophageal cancer. Lowers
cauliflower, kale, turnips, the risk of gastrointestinal
Brussels sprouts, cabbage, cancer.
radish and turnip.
Sulforaphane Found in cruciferous Reduces the risk of breast cancer
vegetables such as broccoli, and prostate cancer.
cauliflower, cabbage and
kale. The richest source of
suHoraphane is broccoli
sprouts.
Proanthocyani skin and seeds of grapes Acts against highly metastatic
dins breast carcinoma.
Quercetin Found in apple, tea, onion, Reduces the cancer risk of
nuts, berries, cauliflower and prostate, ovary, breast, gastric
cabbage. and colon cells.
184
Apoptosis
A flavonol of red grapes, Reduces the cancer risk of

Resveratrol blueberries, mulberries and prostate, ovary and breast
peanuts
Lycopene Pigment of ripe tomatoes. Shows an inverse relation ship
between blood lycopene levels
(or tomato consumption) and
cancer risk of prostate, lung and
stomach.
Saponins Best known sources are peas Helps to prevent colon cancer
and soybeans.
Daidzein Soy beans, kudzu, red clover Reduces incidences of prostate
cancers and prevenls mammary
tumours.
Detection of apoptosis:
There are numerous techn.iques available for the detection of
apoptosis. Morphological changes that occur during the process of
apoptosis can be detected by light and electron microscopy.Cell
shrinkage and increase ingranularity are detected by using density-
gradient centrifugation or flow cytometry. Apoptosis can be measured
by utilizing factors such as changes in calcium ion flux, annexin V
binding and transglutaminase (tTG) activity. Some of the common
techniques used to detect apoptosis include PCR analysis, comet assays,
agarose gel analysis, in vitro and in silq. DNA end-Iabeling and ELISA
systems. There is no gold standard for the specific detection of
apoptosis is available.
, TUNEL:
Apoptosis Detections in TdT-mediated dUTP nick end labeling
(TUNEL): TUNEL technique is highly influenced by the extent of
proteolytic digestion. It is used to identify the nuclei in the areas of
necrosis. This method is also used in biparametric analysis and
retrospective studies of cell death. Apoptotic cells of origin can be
identified by biparametric stain. This method labels the free 3'-ends
of DNA by terminal transferase (TNT), and the label is then visualized
by immunohistochemical techniques. TUNEL reaction seems to be
prone to false positive or negative findings and several improvements
of the methods have been proposed. The staining is very dependent
on 1) fixation time of the tissue samples, 2) proteolytic pretreatment
of the section and 3) the concentration of the nucleotides and terminal
transferase used for labeling
185
Comet Assay:
Comet assay is a technique used for measuring DNA strand breaks
in each cell. This technique was first developed by Swedish researchers
Ostlingand Johansson. It is also used to detect DNA damage, in
nutritional research, and genetic toxicology testing.
DNA laddering:
DNA is cleaved at sites located between nucleosomal units, thereby
generating. DNA mono- and oligonucleosomal fragments (180bp
multimers), which may be visualized on agarose gels, but is difficult
to quantify.
ELISA:
Can detects and quantifies cytosolic mononuc1eosomes and
oligonucleosomes Poly ADP-Ribose Polymerase
Annexin:
Annexin V is an anticoagulant protein that preferentially binds
negatively charged phospholipids. An early step in the apoptotic
process is disruption of membrane phospholipid asymmetry,
exposuring phosphatidylserine (PS) on the outer leaflet of the
cytoplasmic membrane. Fluorescently conjugated Annexin V can be
used to detect this externalization of phosphatidylserine on intact
living cells. Propidium iodide is often combined as a second
flurochrome to detect necrotic cells.
Caspase3:
Induction of apoptosis leads to procaspase-3 proteolytic cleavage to
generate an active 18 kDa caspase-3 fragment which then targets key
modulators of the apoptotic pathway including poly-ADP-ribose
polymerase and other caspases, for cleavage.
186
Chapter 3
Stem Cell
Stem cell unique properties:

Stem cells differ from other kinds of cells in the body. All stem cells-
regardless of their source - have three general properties:
1. They are capable of dividing and renewing themselves for long
periods.
2. They are unspecialized.
3. Regulate~ inbinsic signals and the external microenvironmertt
and
4. Can give rise to specialized cell types.
5. Are very few in population.
Self-regeneration is the ability of stem cells to divide and produce
more stem cells. During early development, the cell division is
symmetrical i.e. each cell divides to gives rise to daughter cells each
with the same potential. Later in development, the cell divides
asymmetrically with one of the ~aughter cells produced also a stem
cell and the other a more differentiated cell.
Depending on differentiation potentiality,stem cells are of three types
1. Pluripotent- (can give rise to all the cells of three germ layers)
i.e. Embryonic Stem Cell (ESC)
2. Multipotent- (can transdifferentiate into few types of specialized
cells) i.e. Adult Stem Cells (HPSC, BMSC etc.)
3. Unipotent -(can only differentiate into one types of specialized
cell) i.e Cardiac Muscle Stem Cell.
187
Different developmental potentialities of cells:

Differentiation Number of Example of Cell types resulting from
Potential cell types stem cell differentiation
Zygote
(fertilized
Totipotential All All cell types
egg),
blastomere
All except
cells of the Cultured Cells from all three germ
Pleuripotential
embryonic human ES cells layers
membranes
skeletal muscle,cardiac
Hematopoietic
Multipotential Many muscle, liver cells, all
cells
blood cells
5 types of blood cells
Myeloid (Monocytes,
Oligopotential Few
precursor macrophages, eosinophils,
neutrophils, erythrocytes)
Cartilage cells, fat cells,
Mesenchymal
Quadripotential 4 stromal cells, bone-
progenitor cell
forming cells
Glial-restricted 2 types of astrocytes,
Tripotential 3
precursor oligodendrocytes
Bipotential
precursor from
Bipotential 2 B cells, macrophages
murine fetal
liver
Mast cell
Unipotential 1 Mast cells
precursor
Terminally
differentiated
Nullipotential None No cell division
cell e.g. Red
blood cell
Stem cells are involved in three processes - development, repair of

adult tissue and cancer.
Totipotent zygote is the ultimate if stem cell which has ability to
produce trophoblast and the embryonic membranes. During
development the 32- to 64-cell stage of zygote is called a blastomere
which sticks together to form a tight ball of cells called a morula.
188
Stem Cell
Each of these cells is totipotential. The next stage is blastocyst which

consists of a hollow ball of cells called as trophoblast cells. In addition
to cell division the development is also characterized by cell
migration. The gastrula is composed of three germ layers i.e. the
ectoderm, mesoderm and endoderm. The outer layer or ectoderm,
middle layer is mesoderm and the inner layer is called endoderm.
The lineage differentiation of these three germ layers
Embryonic Differentiated Tissue
Germ Layer
Endoderm Thymus, Thyroid, parathyroid glands, Larynx, trachea,
lung, Urinary bladder, vagina, urethra, Gastrointestinal
(GI) organs (liver, pancreas), Lining of the GI tract
Lining of the respiratory tract
Mesoderm Bone marrow (blood), Adrenal cortex, Lymphatic tissue
Skeletal, smooth, and cardiac muscle, Connective
tissues (including bone, cartilage), Urogenital system
Heart and blood vessels (vascular system)
Ectoderm Skin, Neural tissue (neuroectoderm), Adrenal medulla,
Pituitary gland, Connective tissue of the head and face,
Eyes, ears
As the developmental precedes, stem cells loose there differentiation
potentiality and gain specific fate by determination process. Cells of
the germ layers are more specialized than the fertilized egg or the
blastomere. Germ layer stem cells give rise to progenitor cells or
precursor cells Zygotic Stem Cell by symmetrical cell division prod uces
two stem cells. But by asymmetrical cell division produces one stem
cell and one progenitor cell. The progenitor cell produces two
differentiated cell by an asymmetrical cell division (Fig-IQ)
Z..'iooii':lrt,iiiii':.oa
..... ...n ...
•. Q.~
iiii............... sc.m..::.ll+ 1Item.,.1I
2· cell stage
,
.zr&01e I~
EmblyOllllstem cell (pleul'lpOlenl)
•.,..r
Germ I~ atem cell (multlpolent)
Lineage OWn •
•
(oIIgopo\lonl)
•
~Ined.""'cd" {lri-at~
T.~ cell (nulllpolenl)
Fig.tO: Types of stem cell division.
189
Embryonic Stem Cell (ESC)

Three types of mammalian pluripotent stem cell lines (Fig-H)
A) Embryonal carcinoma (EC) cells which is derived from testicular
tumours (teratocarcinomas)
B) Embryonic stem (ES) cells which is derived from blastocysts (pre-
implantation embryos)
C) Embryonic germ (EG) cells which is derived from primordial
germ cells of the post-implantation embryo
',---f---TrophabIast
-----t..---- BIa&IocysI caviIy

--+",--- Lumen 01 \lIeNs
Fig.ll: Potentiality of ESC. ECS

Pluripotency can be tested in three independent assays:
1.. Ability to differentiate in vitro in a Petri dish
2. Differentiate into teratomas or teratocarcinomas wheIl. placed in
adult histocompatible or inununosuppressed mice
3. In vivo differentiation when introduced into the blastocoel cavity
of a pre-implantation embryo
• ES and EG cells contribute to every cell type, including the
germline
• EC cells colonise most embryonic lineages, but generally
do not colonise the germline
Factors required for maintaining pluripotency:
A very few of the factors regulating self-renewal of stem cell are
known. Feeder layers in vitro provide some critical factors for
maintenance of undifferentiated state, pluripotency and ESCs
expansion. A fe~der layer provide the factor(s) that suppress
190
Stem Cell
differentiation or promotes self-renewal. Leukaemia inhibitory factor

(LIF) is one of the several factors that has differentiation-inhibiting
activity.
LIF : It is a Neuropoeitic and haematopoietic cytokine that activates a
heterodimeric receptors. IL6 family of cytokine receptors activate the
STAT family of transcription factors to inhibit cell differentiation.
Stem Cell Markers:
Markers are unique in their expression pattern and timing provides a
useful tool for scientists to initially identify as well as isolate stem
cells. Various combinations of markers are required for identifying
and isolating cells for stem cell research.
Embryonic Stem Cell Markers:
Od-4: Oct-4 (also termed Oct-3 or Oct3/4). First and most recognized
marker used for the identification of totipotent ES cells. It isa
transcription factor, that activates gene transcription via a cis-element
containing octamer motif.lt is expressed in totipotent embryonic stem
and germ cells. Expression is required for stem cell self-renewal and
pluripotency. Down- regulation of Oct-4 causes differentiation of
embryonic stem (ES) cells. Oct-4 is one of the master regulator for
pluripotency and lineage commitment.
SSEAs (Stage Specific Embryonic Antigens): There are three SSEAs
(SSEA-l, -3 and - 4. SSEA-l) expressed on the surface of
preimplantation eight cell stage murine embryos and also found on
teratocarcinoma stem cell surface. SSEA-l is also expressed in oviduct
epithelium, endometrium and epididymis, some areas of the brain
and kidney tubules in adult mice. SSEA-3 and -4 are synthesized during
oogenesis and are present in the membranes of oocytes, zygotes and
early cleavage-stage embryos. These SSEAs controlls cell surface
interactions during development. Undifferentiated primate ES cells,
human EC and ES cells express SSEA-3 and SSEA-4, but not SSEA-l.
Undifferentiated mouse ES cells express SSEA-l, but not SSEA-3 or
SSEA-4.
Hematopoietic Stem Cell Markers:
CD34: The cell surface sialomucin CD34. it is expressed on human
bone marrow cells and the most critical marker for hematopoietic
stem cells (HSCs). It plays a significant role in early hematopoiesis.
But resent research showed that HSCs may be CD34+ or CD34-
191
depending on the primitiveness of the stem cells. But still CD34+ cells
are used as HSCs stem cell source for different therapeutic aspects.
ABCG2: ABCG2 is a member of the family of ABC transporters and
expressed mainly on CD34- cells and is down-regulated in committed
hematopoieticprogenitors. It is a determinant of the Hoechst-negative
phenotype of side population (SP) and expressed exclusively in
monkey bone marrow, mouse skeletal muscle, ES cells and on SP
cells. Used for positive selection of pluripotent stem cells from various
adult sources.
Sca-1: Sca-1 (stem cell antigen 1, Ly-6A/E), an 18 kDa
phosphatidylinositol-anchored protein, and the most recognized HSC
marker in mice. Sca-1 can also been used to isolate non-hematopoietic
stem cell population of adult bone marrow, fetal liver and spleen.
CD133: it is a 120 kDa, glycosylated protein and may provide an
alternative to CD34 for HSC selection and ex vivo expansion. CD133
expression is not limited to primitive blood cells, but defines unique
cell populations in non-hematopoietic tissues as well. Human neural
stem cells can be directly isolated by using an anti-CD133 Ab.
Mesenchymal/Stromal Stem Cell Markers:
STRO-1: It is expressed by stromal elements in human bone marrow.
STRO-1 + marrow cells is capable of differentiating into multiple
mesenchymal lineages those include hematopoiesis-supportive stromal
cells with a vascular smooth muscle-like phenotype, adipocytes,
osteoblasts and chondrocytes. STRO-1 Ab is in common practice for
the identification, isolation and functional characterization of human
bone marrow stromal cell precursors.
Neural Stem Cell Markers:
Nestin: It is a class VI intermediate filament protein, that is
predominantly expressed in stem cells of the central nervous system
(CNS). Nestin is used xtensively to identify CNS stem cells form
various nervous system and from in vitro culture. Pancreatic islet
progenitors and hematopoietic progenitors also express Nestin in
certain extent.
p75 Neurotrophin R (NTR): also known as low affinity nerve growth
factor (NGF) receptor. p75NTR+ cells are able to differentiate into
neurons, smooth muscle and Schwann cells in culture. It is used to
identify hepatic stellate cells and mesenchymal precursors.
192
Stem Cell
PSA-NCAM (Polysialic acid-neural cell adhesion molecule): It is a

critical factor for many neural developmental processes. PSA-NCAM,
is the embryonic form of NCAM, and is mainly expressed in the
developing nervous system that retain plasticity
Differences between mouse and human ES cells:
NOTES: Human ES cells form relatively flat, compact colonies. They
can easily be dissociated into single cells. But mouse ES cells form
tighter, more spherical colonies that are more refractory to standard
dissociation methods. Human ES cells grow more slowly and
population doubling time -36 hours as compared to less than 22 hours
in case of mouse.
Properties of hESC and mESC cells:
Human and nonhuman primate ES cells share a similar morphology
that is distinct from human EG cells and mouse ES cells. Human ES
cells form relatively flat, compact colonies that easily dissociate into
single cells in trypsin or in Ca+ 2- and Mg+2-free medium, whereas
human EG cells form tight, more spherical colonies that are refractory
to standard dissociation methods, but which more closely resemble
the morphology of mouse ES cell colonies. Moreover, human ES cells
grow more slowly than mouse ES cells; the population-doubling time
of mouse ES cells is -12 hours, whereas the population-doubling time
of human ES cells is about 36 hours.
Paralleling these differences in cellular morphology, human ES cells
differ from their murine counterparts with regard to cell-surface
antigen phenotype. Like undifferentiated primate ES cells and human
EC cells, human ES cells express stage-specific embryonic antigens 3
and 4 (SSEA-3 and SSEA-4), high molecular weight glycoproteins TRA-
1-60 and TRA-1-81, and alkaline phosphatase. Undifferentiated mouse
ES cells do not express SSEA-3 or SSEA-4, but do express the
lactoseries glycolipid SSEA-l, which is not expressed in human ES
cells, rhesus ES cells, or human EC cells. The functional significance
of these antigens is unknown.
Human ES cells also differ from mouse ES cells in their in vitro culture
requirements for undifferentiated growth. Mouse ES cells require
leukemia inhibitory factor (LIF) for undifferentiated proliferation. In
contrast, LIF alone is not sufficient to prevent differentiation of human
ES cells in vitro. Instead, continued undifferentiated propagation of
human ES cells currently require feeder layers and either the presence
of serum or, if cultured in serum-free medium, bFGF. Under
193
conditions of low cell density, human ES cell lines are more difficult
to propagate in serum, with a cloning efficiency of approximately
0.25%. In contrast, culture in both serum replacement medium and
supplemental bFGF significantly increases the cloning efficiency over
culture in serum alone. Fibroblast feeder layers are currently required
to prevent differentiation of human ES cells. How undifferentiated
proliferation can be sustained in the absence of feeder cells is an area
of active investigation. The critical factors produced by fibroblast
feeder layers, which prevent differentiation of human ES cells, are
entirely unknown. Further work is clearly needed to clarify the
mechanisms involved in sustaining human ES cell proliferation,
including specific receptor-ligand interactions, downstream signaling
events, and cellular target molecules. Ultimately, it would be essential
to establish feeder-independent culture conditions, which permit
large-scale propagation of human ES cells in culture.
Human ES cells have demonstrated remarkably stable karyotypes.
Human ES cell lines demonstrate normal XX and XY karyotypes, similar
to ES cell lines from other species, but distinct from human EC lines
derived from teratocarcinomas. This characteristic makes human ES
cells more relevant as a model for the study of developmental biology
mechanisms and for derivation of differentiated cells for
transplantation therapy.
Human ES cells express high levels of telomerase. The expression of
telomerase, a ribonucleoprotein that adds telomere repeats to
chromosome ends, thereby maintaining their length, is highly
correlated with immortality in human cell lines. Most diploid somatic
cells do not express high levels of telomerase and enter replicative
senescence after a finite proliferative life span in tissue culture, usually
after 50-80 populationdoublings. Unique among normal somatic cells,
some populations of adult stem cells (i.e., hematopoietic stem cells) in
vivo also constituitively express telomerase; however, telomerase
activity is not sustained when cells are placed in culture. In contrast,
cells of the early embryo have high telomerase activity levels.
Likewise, human ES cell lines exhibit high telomerase activity levels
even after more than 300 population doublings and passage for more
than 1 year in culture. In summary, properties of cells of the early
embryo, such as normal karyotype and high telomerase activity, are
sustained for an extended period of time by human ES cell lines in
culture. This unique property among human cell lines has important
implications as a tool to study cellular senescence and mechanisms of
stem cell renewal.
194
Stem Cell
Multi lineage differentiation of ESC VI vivo

Although PS cells can differentiate to multiple embryonic and adult
cell types in vitro, pattern formation or organogenesis does not occur
to a significant degree. Differentiation in the context of an in vivo
environment, such as following injection into a host blastocyst or
implantation into mice, unveils the full developmental potential of
undifferentiated PS cell lines. In this context, many of the normal
features of tissue architecture are reproduced. For example, epithelia
exhibit polarity, are enveloped by a basement membrane, and are
surrounded by mesenchyme; complex tissue structures such as hair
follicles, teeth and gut are also formed. Human PS cells injected into
severe combined immunodeficient mice form benign teratomas, with
advanced differentiated tissue types representing all three EG layers.
Easily identifiable differentiated cells in human PS cell teratomas
include smooth muscle, striated muscle, bone, cartilage, fetal
glomeruli, gut, respiratory epithelium, keratinizing squamous
epithelium, hair, neural epithelium, and ganglia. Compared with
human EC cell lines, human ES cell lines exhibit both more advanced
and more consistent developmental potential. For example, the human
EC cell line NTERA2 cl.Dl injected into immunocompromised mice
forms teratocarcinomas containing simple tubular structures
resembling primitive gut, neural rosettes, and tissue resembling
neuropile
Embryonic inductive events and complex epithelial-mesenchymal
interactions control the formation of organized tissue structures
during normal embryogeneSis. These events and interactions begin
to occur in teratomas but are less pronounced during in vitro
differentiation. Unfortunately, the precise inductive events regulating
embryonic pattern formation are still being elucidated and cannot
yet be reliably reproduced in vitro.
Because in vivo differentiation of human ES cells is more complete
than in vitro differentiation, it would be useful to explore means to
extract the cells or tissue of interest from the heterogeneous mix of
tissues comprising teratomas or to direct differentiation in vivo to a
particular lineage.
Multi lineage differentiation of ESC in vivo:
When removed from feeder layers and transferred to suspension
culture, ES cells begin to differentiate into multicellular aggregates of
differentiated and undifferentiated cells, termed embryoid bodies
195
(EBs), which resemble early post-implantation embryos. Human EBs

frequently progress through a series of stages beginning as simple,
morula-like EBs eventually forming cavitated and cystic EBs between
7 and 14 days of post-differentiation development As for mouse and
nonhuman primate ES cells, differentiation in vitro is consistently
disorganized and frequently variable from one EB to another within
the same culture. A more comprehensive understanding of the
morphology of human EBs and the relationships among different cell
types comprising these complex embryo-like structures may yield
important new information on early inductive events in human
development (Fig-12)
EcIDderm
Skin cells
ut
epidllrmls
Neuron
of brain
"'Piplllt
... cell
Red
blood
cells
Smooilt
rooscle
[111 gut)
Fig.12: In vivo differentiation of ESC.

Human ES cells, like nonhuman primate ES cells, are able to
differentiate into trophoblast in culture. Nonhuman primate ES cell
lines spontaneously differentiate in vitro into extraembryonic
endoderm lineages, including yolk sac, and into trophoblast, as
evidenced by -fetoprotein and chorionic gonadotropin (CG) mRNA
synthesis, and bioactive CG production. Similarly, human EBs
synthesize -fetoprotein transcripts and secrete -fetoprotein and hCG
into the culture medium. Human ES cells, therefore, represent a useful
model in which to study human placental development and function
196
Stem Cell
In vitro differentiation:
Mouse ES cell lines are able to differentiate in vitro into a variety of
embryonic and adult cell types fromall three EGlayers. These include
cardiomyocytes, hematopoietic progenitors, yolk sac, skeletal
myocytes, smooth muscle cells, adipocytes, chondrocytes, endothelial
cells, melanocytes, neurons, glia, pancreatic islet cells, and primitive
endoderm. From these experiments it is clear that ES cells induced to
differentiate in culture follow many of the critical developmental
stages found in the normal embryo and are ultimately able to generate
post-mitotic terminally differentiated cell types depending on the
particular growth factor conditions.
As a result of their ability to differentiate into many different cell
types, ES cells have been recognized as a valuable model system for
studying the mechanisms underlying lineage specification during the
early stages of mammalian development. For example, by comparing
downstream gene expression profiles between null mutant and wild-
type ES cells, one can dissect the complex network of transcription
factor genes regulating tissue-specific gene expression. Also, in vitro
culture provides a unique setting enabling control of the extrinsic
l
Human . • ISOlated ,met cell !i16$!1
emtlr)OOC
Slem cells" l FeedEw layer
I
Embryoid bodies
Derivation of a human embryonic stem cell line. and differentiation strategies
Fig.13: In vitro differentiation of ESC.
197
cytokine or growth factor environment to study how these factors

influence cellular differentiation. Furthermore, in vitro differentiation
of ES cells transduced with gene trap vectors can be used to discover
novel developmentally regulated genes that are important in tissue-
specific differentiation programs. thus, developmental pathways of
cell lineages, which can be derived from ES cells, can be studied using
this in vitro model system (Fig-13)
Recent studies demonstrate that human ES cells differentiating in
culture are able to activate the expression of genes restricted to each
of the three EG layers. Human EBs derived from the human ES cell
line, H9, transcribed genes for -fetoprotein, neurofilament 68kDa
subunit, -globin, and -cardiac actin marking primitive endoderm,
neuroectoderm, and mesoderm derivatives' Differentiating cells
acquired morphologies characteristic of neurons and cardiomyocytes.
Possible methods of achieving in vitro differentiation of human ES
cells
• adding specific combinations of chemical morphogens or growth
factors;
• co-culturing or co-transplanting ES cells with inducer tissues or
cells;
• implanting ES cells into specific organs or regions of animals;
• overexpressing tissue-specific homeobox transcription factor
genes;
• selecting cells that activate a particular lineage-specific gene
expression program
• Isolating cells of interest based on fluorescence-activated cell
sorting
Adult stem cells:
An adult stem cell is an undifferentiated cell found among
differentiated cells in a tissue or organ, can renew itself, and can
differentiate to yield the major specialized cell types of the tissue or
organ. The primary roles of adult stem cells in a living organism are
to maintain and repair the tissue in which they are found. Some
scientists now use the term somatic stem cell instead of adult stem
cell. Unlike embryonic stem cells cells, which are defined by their
origin (the inner cell mass of the blastocyst blastocyst), the origin of
adult stem cells in mature tissues is unknown.
198
Stem Cell
Trans differentiation: it is the capacity of se to be differentiated or

to take in different fate as described in the following figure-14
Adult Stein Cell Uses
...
Done Marrow
(-.l'Orna.....
MUlTOW
Bone
H.~nc:hyl1lOl")
...
Peripheral Olood
Bone Marrow
aloud cells
...
O .... ln
Brain
Nerves
CartUaae
Tendon
Muscle
Pm
Llv_
BrainlNerve
...
Skeletal MU"~'e
Skeletal muscle
Smooth muscle
Blood cell.
Skin (epiderm")
Corned
H • • rt
Blood_lis Bono IJiver
Cartl1aae Lun.
Pat
H_rt
Fig.l4: Trans-differentiation of adult stem cell.

Differentiation pathways of adult stem cells:
Hematopoietic stem cells give rise to all the types of blood cells: red
blood cells, B lymphocytes, T lymphocytes, natural killer cells,
neutrophils, basophils, eosinophils, monocytes, macrophages, and
platelets. Bone marrow stromal cells (mesenchymal stem cells cells)
give rise to a variety of cell types:bone cells (osteocytes), cartilage
cells (chondrocytes), fat cells (adipocytes), and other kinds of
connective tissue cells such as those in tendons. neural stem cells in
the brain give rise to its three major cell types: nerve cells (neurons)
and two categories of non non-neuronal cells - astrocytes and
oligodendrocytes. Epithelial stem cells in the lining of the digestive
tract occur in deep crypts and give rise to several cell types: absorptive
cells, goblet cells, Paneth aneth cells, and enteroendocrine cells. Skin
stem cells occur in the basal layer of the epidermis and at the base of
hair follicles. The epidermal stem cells give rise to keratinocytes, which
migrate to the surface of the skin and form a protective layer. The
follicular stem cells can give rise to both the hair follicle and to the
epidermis.
Developing Transplantation Therapeutics:
Diseases that result from the destruction and/ or dysfunction of a
limited number of cell types, such as diabetes mellitus, in which
pancreatic islet cells have been selectively destroyed, or Parkinson's
199
disease, which results from the destruction of dopaminergic neurons

within a particular region of the brain, could be treated by the
transplantation of differentiated derivatives of ES cells (Fig-1S). Studies
in animal models show that transplantation of either pluripotent stem
cell derivatives, or fetal cells, can successfully treat a variety of chronic
diseases, such as, diabetes, Parkinson's disease, traumatic spinal cord
injury, Purkinje cell degeneration, liver failure, heart failure,
Duchenne's muscular dystrophy, and osteogenesis imperfecta
Fig.IS: Cell based therapeutics.

Although considerable progress in human transplantation medicine,
the chief obstacles in widespread application of cellular transplantation
that face this field are -
• The need for massive doses of immunosuppressive drugs to
prevent rejection of the transplanted tissue and the scarcity of
organs from human cadaver donors. In light of these obstacles,
a human ES cell-based strategy could permit the generation of
an unlimited supply of cells or tissue from an abundant,
renewable, and readily accessible source. Moreover, by virtue
200
Stem Cell
of their permissiveness for stable genetic modification, ES cells

could be engineered to escape or inhibit host immune responses.
The first step toward successful development of a stem cell-based
therapy for human diseases is to establish that human ES cells are
capable of differentiating to a particular cell type of interest and to
purify this lineage from the mixed population. Unfortunately, the
heterogeneous nature of development in culture has hampered the
use of ES cell derivatives in transplantation studies.
To overcome this
• Use of a tissue-specific promoter to drive a selectable marker
such as an antibiotic resistance gene.
• Transduction of a gene construct containing a tissue-specific
promoter / enhancer controlling expression of a green
fluorescence protein gene.
In this way, cells activating a lineage differentiation program of
interest could be selected by fluorescence-activated cell sorting in
much the same way that CD34+ hematopoietic stem cells are selected
and sorted for stem cell transplantation.
• A third major milestone on the road to clinical trials will be to
demonstrate efficacy in rodent and large animal models of
disease.
Rhesus ES cells and the rhesus monkey provide an excellent preclinical
model for developing ES cell-based transplantation therapies and for
testing strategies to prevent immune rejectionlndeed, for Parkinson's
disease and diabetes mellitus, good models are already available in
the rhesus monkey. Replacment of infarcted heart muscle or scar tissue
with ES cell-derived cardiomyocytes will require that new muscle
cells integrate with the existing muscle, contract in a coord,inated and
mechanically useful manner, and develop a new blood supply.
Although complex structural integration would be essential for some
cell transplants (e.g., neurons and cardiomyocytes), normal functioning
of other ES cell-derived transplants will be more independent of such
complex tissue interactions (e.g., islet cells and hematopoietic cells).
• Fourth, the possibility arises that transplantation of differentiated
human ES cell derivatives into human recipients may result in
the formation of ES cell-derived tumors.
Preventing Immunologic Rejection of Transplanted Cells
A fifth consideration is the prevention of immune-mediated rejection
of the human ES cell-derived cellular graft. Currently used
201
immunosuppressive drugs are associated with numerous complications

including wound healing, opportunistic infections, drug-related
toxicities, skin malignancies, and low-grade lymphomas called post-
transplant lymphoproliferative disorders. Instead, human ES cells
could be genetically manipulated to reduce or eliminate immune-
mediated rejection so that lifelong pharmacologic immunosuppression
would not be required.
One potential method for limiting the immune response is to decrease
the immunogenicity of transplanted cells. HomologouS"ecombination
has been used to "knock-out" major histocompatibility complex (MHq
class I and class 11 molecules in mouse ES cells. But, MHC class 1- and
class II-deficient skin grafts are still rejected, possibly on the basis of
indirect allo-recognition-mediatedrejection and/ or natural killer cell-
mediated destruction. Thus, in addition to deleting foreign MHC
genes, it might be necessary to "knock-in" the desired MHC genes,
so that ES cell-derivative transplants are seen as "self" by the
prospective recipient.
Alternatively, genes for immunosuppressive molecules such as Fas-
ligand could be inserted into ES cells, or important immune-
stimulating proteins, such as B7 antigens or CD40-ligand, could be
deleted from ES cells. Irrespective of the method used, the ability to
sta~ly integrate genetic modifications into ES cells provides an
advantage over using adult somatic cells, which are less reliably
genetically altered.The MHC expression profile of human ES cell
derivatives will depend on the degree of differentiation and/ or the
specific cell type derived. For example, adult somatic cells normally
only express MHC class I antigens, whereas B cells, macrophages,
and dendritic cells typically express both class I and class 11 antigens.
Furthermore, whereas most adult organs and tissues harbor
immunostimulatory dendritic cells and vascular endothelial cells, these
normal tissue components would be expected to be absent from ES
cell-derived cellular or tissue transplants. Following figure -16 shows
the strategy for stem cell therapy.
The specific removal of antigen-presenting cells from solid organ
transplants generally increases graft survival. Consequently, some
ES cell-derived tissues may be rather inert immunologically, while
others, like hematopoietic stem cells, may be as immunogenic as normal
adult tissues. Therefore, human ES cell-derived transplants may in
some cases provide an inherentimmunologic advantage compared to
human cadaver tissue transplants.
202
SfemCell
Human ES cells
• Unaage I18Iection by cell survival

Establish pure cultures or oeIllOrting (e.g., IneuIln pnlmoter
of specific cell type drivtng antibiotic resi8W1Ce gene
orGFP)
• Induce with supplemental growth

IaeIor(s) or inducer cella
(e.g .• retinolc acid for neural cells)
Test physiologic
function
----------=1":"''"'---------
,-_.D.ee.mffi.~_s:_te_ _.. 1I De=~ate I Test methods
to prevent rejection
• In rodent models • In non-human pnmate model with • Multi-drug tmrnunoaupp.-ion

mesus ES cell-denved tiSSUes
• In non-human pnmate model • Create dlfferenbated cells
with rheaua ES celklerilled cells • Show absanc:a of tumor formation Ieogenic: ID prospect,ve recipient
(e.g.. dlabell88 and Par1cInson'a USing nlldear repnlgrammltlg
d,_ models in primatae) • Show absenc:a of transmission
of infectious agents • Trensduce ES cells ID express
• Evaluate Integration ",ID heel reelplent MHC genes
tl8sue (e.g•• c:ard.omyocytae for
I /
t_men! of heart failure) • Establish hematopoietlc chimera
and ImmunotoglC fIlleranee
• ? recurrent aulOimrnuntly (e.g., dlabetea)
Human trials I
Fig. 16: Strategy for stem cell therapy.
Nuclear transfer technology may provide a more precise means to

prevent rejection of transplanted cells. Since all nuclear genes here,
including major and minor histocompatibility loci, would be seen as
11 self." .Establishing hematopoietic chimerism is another potential
means of preventing rejection of transplanted cells. This relatively

mild treatment can permit long-term engraftment and could
potentially allow successful solid organ transplantation in humans
without prolonged immunosuppressive therapy. By using the same
ES cell lines to derive both hematopoietic stem cells and other lineages,
it may be possible to initially achieve hematopoietic chimerism followed
by engraftment of a second cell type.
203
Bone marrow-derived stem cells:

Bone marrow (BM) contains hematopoietic stem cells (HSCs), which
differentiate into every type of mature bloqd cell; endothelial cell
progenitors; and marrow stromal cells, also called mesenchymal stem
cells (MSCs), which can differentiate into mature cells of multiple
mesenchymal tissues including fat, bone, and cartilage. Recent findings
indicate that adult BM also contains cells that can differentiate into
additional mature, nonhematopoietic cells of multiple tissues including
epithelial cells of the liver, kidney, lung, skin, gastrointestinal (GI)
tract, and myocytes of heart and skeletal muscle (Fig-l7).
Types of marrow-derived stem cells:
A) Hematopoietic stem cells:
The only true assay for the presence of hematopoietic stem cells (HSCs)
is their ability to reconstitute the hematopoietic system of a
myeloablated host as the hematopoietic reconstitution requires
extensive self-renewal of the transplanted HSCs and their
differentiation into every mature blood cell type.
In humans, the CD34+CD38- cell population is enriched for HSCs in
humans. Side population (SP) cells are also enriched for HSCs. They
are called SP cells because they have a unique ability to extrude Hoechst
dye and because, when examined by fluorescence-activated cell sorter
(FACS) analysis, they fall within a separate population to the side of
the rest of the cells on a dot plot of emission data in the blue rather
than the red spectrum. SP cells express the ABCG2 transporter, a
transmembrane protein, which allows them to actively exclude
Hoechst dye and fluoresce in this specific manner. SP cells are also
present in other tissues, including skeletal muscle. Data are conflicting;
some suggest that SP cells can be tissue-specific stem cells within these
organs and others suggest that they are actually bone marrow (BM)-
derived SP cells lodged within these tissues. A single cell provides
long-term hematopoietic reconstitution20% of the time when injected
into lethally irradiated recipients.
B) Marrow stromal cells:
Marrow stromal cells (MSCs) derived in different laboratories using
different techniques share 2 features: growth in culture as adherent
cells with a finite life span and ability to differentiate into osteoblasts,
chondroblasts, and adipocytes in response to appropriate stimuli. One
of the main hindrances to our understanding of the full potential of
204
Stem Cell
MSCs has been confusion in the literature regarding what specifically

defines an MSC and how it should be isolated and grown in vitro. A
wide array of cytokines (eg, fibroblast gtowth factor 2 [FGF2], FGF4,
platelet-derived growth factor-BB [PDGF-BB], leukemia inhibitory
factor) and isolation techniques (eg, immunomagnetic and physical)
have been used to identify and expand MSCs. Furthermore, no specific
constellation of surface markers has been agreed on for these cells.
Some of the surface antigens reported to be on these cells are Stro
1,CD13, -integrins (CD49a and CD49b), l-integrins (CD29), CD44
(hyaluronate), CD71 (transferrin), CD90 (thy-I), CDI06 (vascular cell .
adhesion molecule-l [VCAM-l]), and CD124 (interleukin-4 [IL-4]
receptor). MSCs uniformly lack antigens such as CD45 that typically
identify hematopoietic cells.
Depending various isolation techniques and culture media used to,
MSCs can differentiate in vitro into neuronal-type cells,
Fig.17: in vitro differentiation of BMSC.

205
C) Multipotent adult progenitor cells:

A population of highly plastic, adult-derived BM cells, referred to as
multipotent adult progenitor cells (MAPCs), can be grown in vitro
from postnatal marrow (and other organs) of mice, rats, and
humans.They copurifyinitially with MSCsand grow as adherent cells
in vitro. However, unlike MSCs, MAPCs can be cultured indefinitely
in a relatively nutrient-poor medium. Specific changes in growth
factors induce differentiation of MAPCs into cells bearing endodermal,
mesodermal or ectodermal markers. MAPCs also display their broad
differentiation potential in vivo. For these assays, MAPCs were
derived from ROSA 26 mice, which express -galactosidase under a
ubiquitous promoter so that cells from these animals can be tracked.
ROSA-26-derived MAPCs injected into murine blastocysts resulted
in chimeric mice with ROSA-26 cells contributing to nearly all somatic
tissues, including brain, lung, myocardium, liver, intestine, and kidney.
After intravenous administration into a sublethally irradiated
immunodeficient mouse, MAPCs differentiate to varying degrees into
hematopoieticcells in the marrow, blood and spleen and into epithelial
cells in liver, lung, and intestine. Consistent with the ability to grow
indefinitely in vitro and to self-renew in vivo, MAPCs express
telomerase; telomere length is maintained after many cell doublings.
It is not yet clear whether MAPCs are a distinct, rare subpopulation
of MSCs normally present in humans or whether their in vivo potential
is a phenomenon developed under unique in vitro cell culture
conditions. Effortsto isolate naive, uncultured MAPCs are under way.
In either case, the discovery of these highly plastic cells opens many
promising new avenues of research.
Bone marrow plasticity:
BM can be transdifferenciated into various cell and type of tissues
Source tissue MuItilineage differentiation potential
Bone marrow (BM) Astrocyte, Neuron, Cardiomyocyte, Chondrocyte,
Hepatocyte, Mesangial cell, Muscle, Neuron,
Osteoblast, Stromal cell, Brain (neurons and
macroglia), Liver, Cardiac myocytes, Kidney,
Igastrointestinal tract, Skin, lung, pancreas
BM (MAPC) lung, Blood, Skin, gastrointestinal tract, Liver
BM (KTLS) Purkinje neuron, Liver
MSC lung, Neuronal cells
Circulating cells Cardiac myocytes
206
Stem Cell
Muscle Adipocyte, myotubes, osteocyte, Endothelial cell,

neuron, Chondrocyte, Osteocyte
Trabecular bone Adipocyte, chondrocyte, osteoblast
Dermis Adipocyte, chondrocyte, muscle, osteoblast
Adipose tissue Chondrocyte, muscle, osteoblast, Stromal cell
Periosteum Chondrocyte, osteoblast
Pericyte Chondrocyte, Osteoblast
Blood Adipocyte, fibroblast, osteoblast, osteoclast
Synovial Adipocyte, chondrocyte, muscle, osteoblast
membrane
Arthritis Res Ther. 2003; 5(1): 32-45.
Adult mesenchymal stem cells and cell-based tissue engineering

Sources of mesenchymal stem cells:
Examples of these tissues include adipose tissue, periosteum, synovial
membrane, muscle, dermis, pericytes, blood, bone marrow and most
recently trabecular bone. Currently, bone marrow aspirate is
considered to be the most accessible and enriched source of MSCs,
although trabecular bone may also be considered an alternative source,
Trans-differentiation of MSC:
Adult mesenchymal stem cell (MSC) can differentiate into
chondrocytes, osteoblasts, adipocytes, fibroblasts, marrow stroma,
and other tissues of mesenchymal origin. Interestingly, these MSCs
reside in a diverse host of tissues throughout the adult organism and
possess the ability to 'regenerate' cell types specific for these tissues
(Fig-1S)
Bone marrow contains three main cell types
1. Endothelial cells,
2. Hematopoietic stem cells, and
3. Stromal cells.
Characteristics of mesenchymal stem cells
MSCs are multipotent for their ability, to trans differenciate into a
variety of different cells/tissue lineages
207
o.....tt •• , ...
I~~I
,
I~I
Fig.18: Trans-differentiation of MSC.

Advantages of MSC
1. long-term culture expansion,
2. MSCs display finite life spans, uncharacteristic of immortalized
'stem' cells.
3. full multilineage differentiation potential
Characterization of human MSCs
MSCs isolated directly from bone marrow are positive for CD34.
Isolation and enrichment of the MSC population has been greatly
facilitated by the Stro-1 monoclonal antibody. Marrow-derived MSCs
express SH2, SH3, and SH4 antigens those are absent from osteocytes
and osteoblasts MSCs from human bone marrow constitutively
expresses cytokines, such as granulocyte-colony stimulating factor,
stem cell factor, leukemia inhibitory factor, macrophage-colony
stimulating factor, and IL-6 and IL-11 is consistent with the ability of
208
Stem Cell
MSCs to support hematopoiesis and provide factors that regulate the

marrow milieu itself.
Applications of mesenchymal stem cells in tissue engineering and
regenerative medicine
Bone: Osteogenic differentiation is used for bone regeneration.
Regulation of osteogenic differentiation: Treatment with the
synthetic glucocorticoid dexamethasone stimulates MSC proliferation
and supports osteogenic lineage differentiation. Organic phosphates,
such as a.-glycerophosphate, also support osteogenesis by playing a
role in the mineralization and modulation of osteoblast activities.
ascorbic acid phosphate and 1,25-dihydroxyvitaminD3, are commonly
used for osteogenic induction, with the latter involved in increasing
alkaline phosphatase activity in osteogenic cultures and promoting
the production of osteocalcin. Application of BMP-2 and basic
fibroblast growth factor increases MSC osteogenesis both in vivo and
in vitro
Bone tissue engineering: The use of natural and synthetic biomaterials
as carriers- for MSC delivery has shown increasing promise for
orthopaedic therapeutic applications, especially bone formation.
Porous ceramics of hydroxyapatite and a-tricalcium phosphate loaded
with autologous MSCs for this purpose. These constructs are capable
of healing critical-s~ed segmental bone defects not capable of being
healed by resident cells or by the addition of the osteoconductive
device alone.
Cartilage: Joint pain is a major cause of disability, results from damage
to the articular cartilage by trauma or degenerative joint diseases
such as primary osteoarthritis. Regeneration of cartilage tissue using
autologous MSCs, thereby obviating any donor-site morbidity as is
seen with current repair methods.
Regulation of chondrogenic differentiation: High-density pelleting
and growth in serum-free medium containing specific growth factors
and supplements. The TGF-a superfamily of proteins and their
members, such as the bone morphogenetic proteins (BMPs), are well-
established regulatory factors in chondrogenesis. Another TGF-a
family member, BMP-6, appears to increase the size and weight of
pellet cultures and to increase the amount of matrix proteoglycan
produced. BMP-2 and BMP-9 have also been used in three-dimensional
MSC culture systems
209
Cartilage tissue engineering: MSC-based repair of full-thickness

articular cartilage defects has been attempted in animal models, using
various carrier matrices. Natural polymers such as collagen have
shown promise in early applications. Autologous MSCs dispersed in
a collagen-type-I gel, can repair full-thickness defects on the weight-
bearing surface of medial femoral condyles.
Soft tissues:
Tendon: MSC can differentiated into other connective tissues, such
as muscle, tendons, and ligaments is also being investigated. For
tenogenesis, key factors include culture conditions, growth factors,
and physical stimulation, such as mechanical loading.
Adipose tissue: In vitro adipogenic induction requires specific medium
supplementations, including dexamethasone and 3-isobutyl-l-
methylxanthine. Indomethacin, a nonsteroidal anti-inflammatory
drug, binds to and activates the transcription factor peroxisome
proliferator-activated receptor gamma (PPAR-a), which is crucial for
adipogenesis. Stromal elements of the marrow, perhaps containing
MSCs, can differentiate into either the osteogenic or the adipogenic
lineage, depending upon micro environmental cues.
MSCs may also be considered for gene therapy applications for the
delivery of genes or gene products. Another intriguing prospect for
the future is the use of MSCs to create' off-the-shelf tissue banks.
Cord blood stem cells:
Umbilical cord is a non conventional and reach source of MSC and
Blood stem cell.
Advantages and Disadvantages of Cord Blood:
Advantages , Disadvantages
• Limitless supply • Unable to obtain additional donor" cells
11
• Available on short notice for for

transplant • leukocyte infusion or second transplant
• No donor attritution compared • Fewer total HPCs due to small volumes
with bone marrow registry • Slower engraftment (return of circulating
• Ethnic diversity easier to • neutrophil and platelet numbers)
achieve
• Painless collection of stem cells
• Large inventory product (high up-front
costs;
• Higher proliferative capacity • units may become outdated" due to
11
• Lower rate of acute graft-vs- changes

host disease • in banking standards
210
Stem Cell
Cord blood over Bone marrow:

Advantages
Umbilical cord blood Bone marrow
0 Speed of availability o Faster engraftment
0 Greater likelihood of suitable match o More experience
0 Decreased transmission of viral diseases
0 Reduced graft versus host disease
0 Ease of scheduling transplant
Disadvantages:
Umbilical cord blood Bone marrow
• Less experience • Lack of donors
• Limited cell dose • Longer search times
• Slower engraftment • More graft versus host disease
Cord blood can be transplanted for the therapeutic purpose & their
Current Applications
Acute Leukemia's Stem Cell Disorders
Acute Lymphoblast Leukemia (ALL) Aplastic Anemia (Severe)
Acute Myelogenous Leukemia (AML) Fanconi Anemia
Acute Biphenotypic Leukemia Paroxysmal Nocturnal
Acute Undiffo?rentiated Leukemia Hemoglobinuria (PNH)
Pure Red Cell Aplasia
Chronic Leukemia's Other Inherited Disorders
Chronic Myelogenous Leukemia (CML) Lesch-Nyhan Syndrome
Chronic Lymphocytic Leukemia (CLL) Cartilage-Hair Hypoplasia
Juvenile Chronic Myelogenous Leukemia Glanzmann Thrombasthenia
OCML) Osteopetrosis
Juvenile Myelomonocytic Leukemia Adrenoleukodystrophy
OMML)
Myelodysplastic Syndromes Plasma Cell Disorders
Refractory Anemia (RA) Multiple Myeloma
Refractory Anemia with Ringed Plasma Cell Leukemia
Sideroblasts (RARS) Waldenstrom's Macroglobulinemia
Refractory Anemia with Excess Blasts Amyloidosis
(RAEB)
Refractory Anemia with Excess Blasts in
Transformation (RAEB-T)
Chronic Myelomonocytic Leukemia
I(CMML)
211
Inherited Immune System Disorders Inherited Metabolic Disorders

Ataxia-Telangiectasia Mucopolysaccharidoses (MPS)
Kosbnann Syndrome Hurler's Syndrome (MPS-lH)
Leukocyte Adhesion Deficiency Scheie Syndrome (MPS-IS)
DiGeorge Syndrome Hunter's Syndrome (MPS-Il)
Bare Lymphocyte Syndrome Sanfilippo Syndrome (MPS-UI)
Omenn's Syndrome Morquio Syndrome (MPS-IV)
Severe Combined Immunodeficiency Maroteaux-Lamy Syndrome (MPS-
(SCID) VI)
SCID with Adenosine Deaminase Sly Syndrome, Beta-Glucuronidase
Deficiency Deficiency (MPS-VII)
Absence of T & B Cells SCID Adrenoleukodystrophy
Absence of T Cells, Normal B Cell SCID Mucolipidosis n (I-cell Disease)
Common Variable Immunodeficiency Krabbe Disease
Wiskott-Aldrich Syndrome Gaucher's Disease
X-Linked Lymphoproliferative Disorder Niemann-Pick Disease
Wolman Disease
Metachromatic Leukodystrophy
Myelodysplastic Syndromes Histiocytic Disorders
Refractory Anemia (RA) Familial Erythrophagocytic
Refractory Anemia with Ringed Lymphohistiocytosis
Sideroblasts (RARS) Histiocytosis-X
Refractory Anemia with Excess Blasts Hemophagocytosis
(RAEB)
Refractory Anemia with Excess Blasts in
Transformation (RAEB-T)
Chronic Myelomonocytic Leukemia
I(CMML)
Myeloproliferative Disorders Inherited Erythrocyte
Acute Myelofibrosis Abnormalities
Agnogenic Myeloid Metaplasia Beta Thalassemia Major
(myelofibrosis) Sickle Cell Disease
Polycythemia Vera
Essential Thrombocythemia
Lymphoproliferative Disorders Inherited Platelet Abnormalities
Non-Hodgkin's Lymphoma Amegakaryocytosis / Congenital
Hodgkin's disease Thrombocytopenia
Phagocyte Disorders Other Malignancies
Chediak-Higashi Syndrome Ewiflg Sarcoma
Chronic Granulomatous Disease Neuroblastoma
Neutrophil Actin Deficiency Renal Cell Carcinoma
ReticularDysgenesis Retinoblastoma
212
Stem Cell
Diseases and Maladies Treated Using Adult Stem Cells

Cancers: Auto-Immune Disease: Spinal Cord Injury
Brain Cancer Multiple Sclerosis Stroke Damage
Retinoblastoma Crohn's Disease Anemias/Blood Conditions
Ovarian Cancer Scleromyxedema Sickle Cell Anemia
Skin Cancer: Merkel Cell Rheumatoid Arthritis Sideroblastic Anemia
Carcinoma
Testicular Cancer Juvenile Arthritis Aplastic Anemia
Tumors Abdominal Systemic Lupus Amegakaryocytic
Organs Lymphoma Thrombocytopenia
Non-Hodgkin's Polychondritis Chronic Epstein-Barr Infection
Lymphoma
Hodgkin's Lymphoma Sjogren's Syndrome Fanconi's Anemia
Acute Lymphoblastic Behcet's Disease Diamond Blackfan Anemia
Leukemia
Acute Myelogenous Myasthenia Thalassemia Major
Leukemia
Chronic Myelogenous Autoimmune Cytopenia Red Cell Aplasia
Leukemia
Juvenile Mye1omonocytic Systemic Vasculitis Primary Amyloidosis
Leukemia
Cancer of the Lymph Alopecia Universalis Wounds/Injuries:
Nodes:
Angioimmunoblastic
Lymphadenopathy
Multiple Myeloma Cardiovascular Limb Gangrene
Myelodysplasia Heart Damage Surface Wound Healing
Breast Cancer Ocular: Jawbone Replacement
Neuroblastoma Corneal Regeneration Skull Bone Repair
Renal Cell Carcinoma Immunodeficiencies: Other Metabolic Disorders:
Various Solid Tumors X-Linked Hyper Osteogenesis Imperfecta
Immunoglobuline-M
Syndrome
Soft Tissue Sarcoma Severe Combined Sandhoff Disease
Immunodeficiency
Syndrome
Waldenstrom's X-linked Hurler's Syndrome
Macroglobulinemia Lymphoproliferative
Syndrome
Hemophagocytic Neural Degenerative Krabbe LeukodystrophY
Lymphohistiocytosis diseases/ injuries
POEMS Syndrome Parkinson's Disease Osteopetrosis
Cerebral X-Linked
Adrenoleukodystrophy
213
Potential Future Stem Cell Applications:

Alzheimer's Disease, Cardiac Disease, Diabetes, Lupus, Multiple
Sclerosis, Muscular Dystrophy, Parkinson's Disease, Rheumatoid
Arthritis, Spinal Cord Injury and Stroke.
Possible Approaches to Stem Cell Therapy for Cerebral Infarction:
Organ-specific stem cells are harvested from the brain, expanded in
vitro, and reimplanted into the patient. New neurons may be derived
from the neural stem cells to replace those lost during infarction,
enabling the patient to regain lost neurologic function. B, Human
embryonic stem cells from allogeneic donors are reprogrammed in
vitro into neural precursor cells and then reimplanted into the patient.
C, Somatic cells (eg, skin cells) are obtained from the patient, and
somatic nuclei are harvested and transferred to .enucleated human
oocytes. A blastocyst is formed from the resulting cell. Cells from the
inner mass of the blastocyst are cultured and reprogrammed in vitro
to create neural precursor cells, which are then used to repopulate
the damaged tissue without risk of immunologic rejection. D, Bone
marrow stem cells are harvested from the patient, reprogrammed in
vitro to become neural precursor cells, and reimplanted into the patient
to repopulate the damaged area. E, Combination therapy in which
bone marrow stem cells are harvested, genetically altered through
gene transduction, reprogrammed in vitro to become neural precursor
cells, and reimplanted into the patient (Fig-19).
Fig.19: Approaches to Stem Cell Therapy for Cerebral Infarction.

214
Chapter 4
Eugenics
Every mental and physical characters of human being are controlled
by genes. Some characters are useful to human and some characters
are harmful to society. One question may arise. Whether some change
is occurring in genetic composition? If it is, in which direction? What
should be done for this problem? Sir Francis Galton was the first
scientist who paid attention to it. Galton agreed with Darwin's theory
of evolution. He stated that like in other animals, human beings also
have process of evolution. Sir Francis Galton in the year 1883 evolved
the word eugenics. He defined it as eugenics is the study of those
factors by which development of mental and physical characters takes
place in human beings. In other words we can say that the branch of
genetics in which the laws of inheritance are used to improve human
beings is called eugenics. Eugenics is a Greek word meaning well
born.
Aims and ideals of eugenics
It is a matter of common agreement that human being is not complete.
In other words there is some mental or physical problems which needs
improvement. Following characters of human being may need
improvement:
1. Health: Mental and physical healths of human being are very
important. From genetics point of view health means that man
should be free from hereditory defects, disease and weakness
which may reduce the work efficiency, happiness or life. Beauty
is a part of good health because good health and beauty have
positive correlation.
2. Intelligence: Meaning of intelligence is the capacity to think and
understand. High level of intelligence is useful to human being
and society.
3. Moral character: It means that human beings should have cordial
and good relationship with the neighbourers and society. He/
she should have high character and excellent behaviour.
215
Genetics: Fundamentals and AppliClltions
4. Variability: To attain specialization in knowledge is very useful

to the person and society. Generally the capacity for specialization
is hereditary: .,
Methods of improving mankind
To improve human being two methods may be used:
First method is euthenics. It means by improving the environment
and surroundings, improvement is brought in human beings. By the
word environment we mean education, nutrition, medical facilities,
dress, house etc.
The second method is eugenics. Under it improvement in hereditary
constitution are studied. This method is of following two types.
1. Positive eugenics: Under positive eugenics studies are made
for production of superior hereditary characters.
2. Negative eugenics: Under this studies are made to get rid of
undesirable germplasms
Positive eugenics:
Positive eugenics is used to develop person with superior hereditary
characters. In addition to it efforts are made to prevent good
germplasm from deterioration. Therefore it is important that persons
of exel1ent heredity and inheritance should produce more children.
Hence it is important that these persons should be provided sound
financial conditions by the government.
Generally it is seen that persons of excellent hereditary characters
which have specialized knowledge and excellent human beings
produce less children. One of the reason is late marriage. This reason
is being discussed below.
Late marriage:
There are several reasons for late marriage. Details and their possible
reasons are mentioned below.
1. Higher education: Higher education is one of the main reason
for late marriage. Boys and girls attaining higher education usually
marry at the age of 30 - 35 years. Reproductive age in man is upto
age of 45 years and in women is upto age of 40 years normaly.
Therefore highly educated men and women get reproductive stage
only for 10 - 15 years .
.On the other hand lower grade persons marry at the age of 17 - 22
216
Eugenics
years. Therefore they get quite a long reproductive phase.

Solution to the problem:
(a) To stop late marriage in higher educated men and women. One
step is to encourage such persons while studying. They should
be provided all financial help for reasonable good living. Beside
this following restrictions & imposition may help further:
(b) There should be restriction for late marriage in highly educated
persons by imposing tax.
(c) Boys and girls attaining higher education and willing for early
marriage must be given scholarship, freeship, free books,
stationary etc. from the government.
2. Reduction in sexual instinct: Persons who pass a long period
of their life for higher education sometimes suffer from temporary
suppression of other psychological feelings. This temporary
suppression some times develop physiological disturbances due to
which their gonads do not function properly. That is the reason that
why high intellectuals sometimes becomes sexually incapable.
Therefore in them the feeling develop for not to marry.
For solution, entertainment must be made a part of higher education
and presence of those high intellectuals must be made compulsory.
By doing so their sexual instinct will be alive and they will marry and
produce children.
3. Poor economic position in early years of employment: Poor
economic situation is also a cause for late marriage. Nowadays there
is competition everywhere including employment. It has been seen
that employment is not given as per the qualification. Therefore a
person who has passed a long part of his life to get higher education
does not get employment for long time, or such person gets
underemployment which does not suit to their qualification. In that
case the person gets disappointed and think that it would be better
to marry only after getting proper job so that it would be possible to
maintain the family properly.
Solution to the problem-Solution can be done by follOwing methods:
a. Persons with high education must get such a suitable & dignified
employment which has high salary as well as respect.
b. Such persons after marriage must be given enougH marriage
allowance.
217
c. After birth of every child the person must get sufficient increase
in pay.
4. High cost of living: All of us know that now a day's price of
every item is increasing. After marriage when the lady is in carrying
stage and maternity stage, that time she requires several items. Due
to high prices it becomes a problem how to procure them. Due to this
reason they tries to postpone the conception till they have sound
financial condition.
In addition to it housing is another problem Reasonable good house
is the requirement of newly married couple. But the price of land and
price of construction is so much high that it becomes extremely difficult
or sometimes impossible.
The problems mentioned above have effect on number of children.
They prefer to have less children so that their expenses can be born.
In addition to the above mentioned problems education of the child
is another problem for high intellectuals. Nowadays standard of
education in government school is poor. Therefore they try to admit
their children in convent school. But admission fee and monthly fee
are exorbitant in convent school. Due to these reasons high intellectual
persons produce fewer children so that they can get them educated
properly.
(a) For delivery government must provide financial help to such
persons.
(b) High intellectuals must get special grant to build a house. They
must get land at reasonably cheap price.
(c) High intellectuals must get maternity allowance.
(d) Such persons must be given additional money for education of
children, otherwise low cost convent school should be provided.
5. Lack of freedom of marriage with desired mate:In Indian society
sometimes does not allow marriage between girls and boys who love
each other. During long period of education sometime boys and girls
of other cast starts love affair. Mostly parents does not allow such
marriages. Due to which those boys and girls of high caliber decide
not to marry.
Solution of the problem:
Government should make a rule that boys and girls of high caliber
218
Eugenics
,
even may be of different cast can marry against their parent's wishes.
Thus it is clear that if above mentioned points are executed properly
then there will be definitely improvement in human race.
Negative eugenics:
For welfare of human beings negative eugenics can be of great help.
Following methods may be used:
1. Sterilization: Undesired persons not fit for the nation should
be sterilized so that they may not produce children.
2. Stopage of child birth in parents of defective inheritance and
avoid marriage between blood relation.
3. Avoid marriage between defective persons.
4. Check on immigration.
These are detailed below:
1. Sterilization:
Sterilization is a good technique for welfare of the country. This should
be applied on undesirable persons to prevent them producing children.
Sterilization is dependent on surgery. Under this in men vasectomy
and in women salpingectomy operation is done. These surgical .
operations are explained below:
Vesectomy:
It is a minor surgery. Doctor open the scrotum and cut the spermduct
upto some length and rest is tied properly. Afterwards scrotum is
stitched. By this surgery the man looses the power of reproduction.
But production of male harmone in testes continues. Though seminal
vesicles and prostrate glands ejaculatory fluid is produced as before
but sperm production stops and thus the man can not produce child.
Sal pingectomy:
Salpingectomy is a complicated surgery. It is done in ladies. In this
stomach is cut. Then the oviduct is cut. The cut ends are tied .
Afterwards the stomach is stiched. After this surgery production of
endocrine harmone continues. So the lady carries on the married life .
as before. However because ovum is not produced so she is unable
to produce child.
In the above mentioned operations male and female have permanent
sterility. However if in males spermduct (vasadiference) and in female
219
oviduct (fallopian tube) are tied with surgical thread which can be
joined when desired, then in both cases sterility will be temporary.
Nowadays sterilization techniques are being widely practiced. United
States of America has also legalised this surgery. Sterilization surgery
is specially recommended in case of undesirable persons such as
idiotic, insane, tuberculosis, cancer, leprocy,etc. If these surgeries are
done in right sense then these hereditary diseases can be eliminated
from the world one day.
2. Stopage of child birth in parents of defective inheritance and
avoid marriage between blood relation:
Different type of sex linked characters for example night blindness
etc. are definitely inheritable which are controlled by dominant genes,
heterozygous recessive condition, sex linked recessive gene and
autosomal recessive gene.
If disease caused by recessive genes occurs in a family, then it may
pass from generation to generation without expression, because
generally husband or wife are normal, but if by chance marriage takes
place between first cousin in that case the children gets this gene in
homozygous recessive stage. So they develop the disease of colour
blindness.
Looking the above mentioned facts it is very important for welfare
of the nation we should avoid marriage between close relatives. Some
advanced countries such as United States of America has abandoned
marriage between close relatives by rule.
3. Avoid marriage between defective persons:
To avoid proliferation of bad germplasm another method is to avoid
marriage between defective & undesirable persons. By doing so the
society will be benefited in two ways viz.(i) persons who are habitual
criminals, beggars, drinkers, mads will not be able to produce children
because their children may become like the parents. (ii) such bad
persons will not be spoiling good persons of the society.
4. Check on immigration of foreigners:
Under negative eugenics the last important suggestion is check on
immigration of foreigners. The reason is that if foreigners settle
permanently then there are chances of transfer of undesirable genes
in Indians. This fact can be understood by an example. Suppose a
foreigner having epilepsy or/and tuberculosis in recessive form if
220
Eugenics
settle in India, he may marry with a women who has recessive genes
for these diseases. In that case in their children both recessive genes
will appear in homozygous stage. The result will be that their children
will express those bad diseases.
Therefore before allowing any foreigner to settle in India it is very
important to find out properly that whether that person has any
disease in heterozygous or homozygous phase.
The above mentioned methods of positive eugenics, negative eugenics
and euthenics if used in good combinations then there will be good
success in improvement of human race.
221
Chapter 5
Plant Breeding
India is an agricultural country. Seventy percent of its population are
farmers. Despite this, food problem is an important problem of our
country. Beside India other countries of the world are facing the same
problem. How to solve this problem? It appears that answer to this
question is quite difficult because population.of the world is increasing
at the rate of over 2.2 crores every year. To solve this problem plant
breeding may help. Therefore it is essential that plant breeders should
develop such improvement in crops so that farmers may reap more
and more yield of better quality.
Plant breeding is a subject which is related to genetics, cytology and
cytogenetics and has its own special features. Plant breeding is based
on principles of genetics and studies in details the technologies of
crop improvement. A good plant breeder has good understanding of
genetics and cytogenetics. In addition to these he/ she should know
agronomy, horticulture, plant pathology, plant physiology, statistics,
agricultural chemistry and entomology.
Plant breeding has been defined by G.M. Poehlman and D.N.
Borthakur (1959) as a science to improve heredity of plants. When by
hybridization between close relatives offspring are produced, it is
known as inbreeding. But if the parents are least related or distantly
related then the breeding is known as out breeding.
Under plant breeding stigma of other plant is artificially pollinated
and offspring are produced for various types of selections. Therefore
plant breeder has to know merits and demerits of the plant and
requirement of the country.
Researc;h stations of plant breeding in India:
In India plant breeding researches are done by state government,
central government and non government agencies. Indian Council of
Agricultural Research (ICAR), Council of Scientific and Industrial
Research (CSIR) and the important central agencies, state agricultural
universities in various states of India are the important state agencies.
222
Plant Breeding
Commercial private seed breeding stations are non gover:nment

agencies. ~reeding research centers in important crops are mentioned
below:
1. Wheat: Main breeding centre on wheat is at Indian Agricultural
Research Institute, New Delhi. Breeding work on this crop is
also being done in various state agricultural universities. Name
of some famous wheat breeders are B.P.Pal, Ram Dhan, B.S.
Kadam, Pathak, Bhatia etc.
2. Rice: There are several breeding centers for rice in various states.
The Central Rice Research Institute is located in Cuttack of Orissa.
In Uttar Pradesh rice breeding centers are in Nagina, Faizabad,
Gorakhpur, Panchperva, Tisuhi etc. Several rice breeding stations
are located in various state agricultural universities of north and
south India. Name of some famous rice breeders are Ramiah,
Kadam, Nagavo, Parthasarthy, etc.
3. Pulse: Breeding researches on pulses are being done in various
states of India. Central Pulse Research Station is located in
Kanpur, Uttar Pradesh. Central Research Station for Soyabean is
located in Madhya Pradesh.
4. Potato: Breeding work on potato is being done at Potato Breeding
Station, Shimla, Himachal Pradesh. Some breeding work is also
being done at Ootachmund (Tamil Nadu), Poona and Shilong.
Name of some famous potato breeders are B.P.Pal, Pushkar Nath,
M.S. SWaminathan, Mundkar etc.
S. Cotton: For cotton breeding work in the country certain
committee have been formed ego Indian Central Cotton
Committee (ICCC). This committee manage cotton breeding
researches done by various states. Central Cotton Research
Institute is located in Nagpur. Name of famous cotton breeders
are Ansari, Avtar Singh, Dastur, Ghose, Ayangar, Jagannath Rao,
Joshi, Kalyan, Raman, Majumdar, Pandey,Lather, etc.
6. Sugarcane: Central Sugarcane Research Institute, Coimbatore is
doing excellent breeding work on this crop. It has a branch at
Lucknow, Uttar Pradesh.
7. Horticultural crops: Indian Institute of Horticultural Research
(IIHR), Bangalore is doing excellent breeding work on fruit crops
such as mango, banana, papaya, guava, citrus, grape, pineapple,
floricultural crops ego rose, marigold, chrysanthemum,
bougainvillea, vegetables ego tomato, brinjal, okra, chilies,
pumpkin, beans, spinach, watermelon,muskmelon,etc.Vegetable
223
breeding is also being done by the Central Vegetable Research

Institute, Varanasi. IIHR, Bangalore is also doing breeding
research on medicinal & aromatic crops such as vinca, khasikateri,
aloe, kevanch, memory plant, jasmines, tuberose, scented
geranium, patchouli, etc. Breeding researches on these crops is
being done by Central Institute of Medicinal and Aromatic Plants,
Lucknow & its regional research stations, National Research
Centre on Medicinal & Aromatic Plants, Anand and various state
agricultural universities.
In addition to above mentioned research centers India has also other
research centers on various other crops such as Indian Coffee Board,
Indian Tea Association, Jute Research Institute, Barrakpur & Neelgang,
West Bengal, Indian Coconut Committee, Kerala, Central Arecanut
Committee, Kerala, Central Tobacco committee, Chennai, and other
research institutes.
Objectives ()f plant breeding:
Objective of plant breeding is to combine various useful characters
into one variety of plant. The useful characters which a plant breeder
must understand are mentioned below:
1. High yield: It is the most important objective of plant breeding.
2. Early maturity: In addition to high yield, early maturity of the
crop is also an objective.
3. High quality:In addition to above mentioned qualities the crop
should be of high quality. It should have sufficient nutrient,
attractive colour, should be tasty and easily digestible. As per
the crop requirement the produce should be of excellent quality.
4. Non shedding of grain: Due to shedding of grain there is
reduction in yield. This problem is more in case of paddy grown
in Kashmir. Therefore plant breeding has the objective to prevent
the shedding. Non shedding varieties of paddy developed are
522 and mtu 7.
5. Resistance to drought and frost: If these objective are met then
the crop can be grown at a place where it was earlier not possible
to grow. For example ground nut (Arachis hypogea) was not grown
in Punjab. However because of development of new varieties
F15 and 5amrala now ground nut can be grown in Punjab.
6. Disease and insect resistance: Plants of vegetables, cereals,
pulses, oil seeds, fruits, wooden plants, medicinal and aromatic
224
Plant Breeding
crops are attacked by several insects and diseases caused by

fungus, bacteria and virus. Everyyear this leads to loss of several
crores rupees in the country. Therefore one objective of plant
breeding is to develop varieties which are resistant to diseases
and insects. For example in case of wheat varieties resistant to
rust are NP 809 and NP 770.
7. More medicinal property: In case of breeding of medicinal crops
the objective is to increase medicinal properties, yield and
resistance to biotic & abiotic stress.
S. More yield of superior quality essential oil: For breeding of
aromatic crops the objective is to increase yield, quality and
resistance to diseases & pests.
In addition to above mentioned objectives a plant breeder makes
efforts to develop varieties which by application of more fertilizers
and manure can produce more yield, the fruits should have uniformly
good size and colour. The crop should have resistance to lodging
and flood.
Methods of plant breeding
Main methods of plant breeding are the followings:
1. Plant introduction
2. Selection
3. Hybridisation
4. Mutation
5. Polyploidy
Before adopting the method of plant breeding the breeder should
know how the plant breeds and propagate. According to method of
reproduction plant can be of two types.
(a) Asexual group: In some crops and fruits the propagation is by
vegetative means such as cutting, layering, budding, grafting,
inarching, tissue culture etc. for example potato, sweet potato,
sugarcane, mango, rose, jasmine, scented geranium, patchouli
etc.
(b) Sexual group: Most of the cereals, pulses, vegetables, some fruits
have sexual reproduction. Under sexual group some crops have
self pollination viz. barley, oat, wheat, paddy, gram, pea etc.
where as some crops have cross pollination system such as maize,
rye, cucurbits, jute etc.
225
Now brief description is being presented for methods of plant

breeding below:
1. Plant introduction:
For improvement of plants, amongst all the methods plant introduction
is the cheapest and also most simple method. Plant introduction means
to procure new varieties and to grow and evaluate. New varieties
are brought from different places in the country or introduced from
different country either by post or by personal visit or by plant
collecting expedition or with the help of Food and Agriculture
Organization. After collection the plants are grown for some
generations at different locations and afterwards tested for general
suitability. By introduction superior variety is produced. In addition
to it this can be included in hybridization programme. Following crop
varieties are introduced in India:
(a) Ridley variety of wheat: It was introduced from Australia. It is
very suitable for hilly tracts of India. It has good yield and
resistant to rust disease.
(b) Tie Shin Tun and FA-17 varieties of potato: These varieties
were obtained from China. FA-17 variety gives high yield and is
quite tasty. Tie Shin Tun variety is quite large in size, attractive
and has high yield.
(c) Sioux variety of tomato: It was obtained from U.S.A. This can
be grown in summer as well as winter. Fruits are of large size
and gives high yield.
(d) Philippine early vari~ty of cowpea: It was introduced from
Philippine. It matures early: The pods are straight, green and
thick. Seeds are red & white and kidney shaped.
(e) Early Badger variety of pea:It was brought from U.S.A. It grows
fast and matures early. Its grains are sweet.
(f) Haryana mulahathi No-1 : It is the Russian collection number
EC 111236 tested and released by CCS Haryana Agricultural
University, Hissar after All India Coordinated Trial.
2. Selection:
Selection means to choose out the most useful variant from a mixed
populaton. One important thing to be taken care is that proper
genotype only to be selected. Selection procedure is of following kinds:
226
Plant Breeding
(a) Mass selection:

From farmer's field (mixed population) good looking useful variants
are selected. Seeds of these selected plants are sown next season or
next year. Again from that population good plants are selected. This
process is repeated for some generations, till the desir~d improvement
is achieved. Mass selection is based on phenotype. Therefore some
times good looking plants are not of good genotype.
(b) Individual plant selection or pure line selection:
Under this method from the mixed population the best plant is
selected and its seeds are obtained. The seeds are sown in next season
or next year. Again from those plants the best plant is selected. This
process is repeated for some generations till the desired improvement
is obtained. By this method qualitative improvement in characters
such as amount of protein, amount of oil, amount of sugar, length of
filament etc. can be possible. This method is based on pureline theory
of Johansan.
3. Hybridization:
By introduction and selection, beyond certain limit, improvement can
not be made. Therefore plant breeder has to adopt suitable breeding
method. In hybridization method good qualities of parents are
combined into offsprings. Hybridization can be defined as cross
breeding between related organisms differing in one or several
characters. The resultant offsprings are called as hybrids.
Hybridization techniques:
Before taking up hybridization the plant breeder has to know the
breeding behaviour of the plant ie. The plant is monoecious or
dioecious, unisexual or bisexual, self pollinated or cross pollinated.
In addition to these the plant breeder must know its floral biology
and following informations.
(1) Flowering period
(2) If the plant belong to gramineae family then what time panicles
emerge.
(3) The sequence of inflorescence.
(4) When the buds bloom.
(5) How many buds bloom each day.
(6) Anthesis time.
227
(7) Dehiscence time.

(8) Receptivity time of stigma.
(9) For how long pollen remains viable.
Now the methods of hybridization are described below.
1. Choice of parent:
In choice of parents care must be taken for objective of breeding.
Then the breeder should decide which parent is to be used as female
and male.
2. Culture of parent plant: The parents should be grown in
experimental garden with optimum growth condition.
3. Protection of pollen: In order to save contamination of pollen
from other plants, anthers should be collected before dehiscence
in clean glass vial. In that vial the anther will dehisce and will be
free from contamination.
4. Emasculation: Removal of anthers before dehiscence is known
as emasculation. The procedure differ from crop to crop. A
needle and forecep are used in this process. The needle and
forecep before use are sterilized by dipping in methylated or
rectified sprit. From hermaphrodite flowers stamens before
dehiscence are taken out by help of needle and forecep. But in
case of monoecious the female flowers have to be covered
carefully In inoculation the petals and sepals should not be
removed unless it is unavoidable.
There is one more technique for emasculation in sorghum crop. The
whole inflorescence is immersed in hot water at 50 centigrade. Due
to this all its pollen grains are killed. Thus imasculation is done.
5. Bagging: After emasculation the inflorescence is covered by
muslin or special paper bag so that outside pollen should not
fall on the stigma. Afterwards a label is tied. On the label is
written the name of female and male parents and date of
pollination This bag should remain there till seed formation is
not complete.
6. Pollination: Pollination should be done at a time when stigma
is receptive. For this purpose the bag should be some what opened
and the collected pollen is applied on the stigma by a brush.
After this process the bag is again dosed properly.
228
Plant Breeding
Difficulties in plant breeding:

During hybridization a plant breeder may have to face following
difficulties:
(1) Maturity period of the parents may be different.
(2) Failure in fertilization may be due to sterility or non crossability.
Favorable conditions for hybridization:
Following circumstances are favorable for hybridization:
(1) Viable pollen
(2) Receptive stigma
(3) Morphological or physiological compatibility between pollen and
stigma.
(4) Resistance of flowers to manipulation.
(5) Chromosome similarity between male and female parents.
(6) Favorable environment for flowering and fruiting.
Hybridization in self pollinated crops:
Self pollinated crops are many viz. oat, wheat, barly, paddy, cotton,
tobacco, peas, beans, etc. In such crops improvement by hybridization
has been classified as mentioned below by Hays, Immer and Smith:
(a) Pedigree selection method.
(b) Bulk method.
(c) Back cross method.
(d) Multiple cross method.
These methods are described below:
(a) Pedigree selection method:
Under this method selection is done in every generation after F2. In
addition to it, breeding behaviour of the selected plant is studied
and proper record is maintained. With this method improvement
takes less time. However this need more labour, care and funds.
Pedigree method is also known as following because of reasons
mentioned against each:
1. Pure line selection: because pureline or inbred line is used.
2. Individual plant selection: because individual plants are the basis
of pedigree selection.
3. Head to row selection: because in thickly planted and profusely
229
tillering crops as wheat and barly where individual plant can

not be selected, head is selected and its seeds are sown separately
in rows.
4. Projeny selection: because on basis of projeny of individual plants
selection for desirable plant is done.
5. Single plant selection: because single plants are selected and their
progenies. are maintained separately.
6. Inbred selection: because inbred or pure line is used.
Differences between clone and pure line are mentioned in following
table:
Table: Differences between clone and pure line.
Points Clone Pureline
Parents Progenies of a simple Progenies of Single self
vegetatively propagated fertilized homozygous
heterozygous plant plants
Production By vegetative means Produced by selfing
Genetical Constitution All members of a clone are All members of pureline
identical having same are identical &
genotype but heterozygous homozygous
Occurrence Occurs in vegetatively Occurs in self pollinated
propagated crops crops
Importance Utilized directly as improved Same
variety as well as parent for
hybridization
(b) Bulk method:
This method matches with mass selection method. Natural
hybridization is allowed in F2 generation to get F3 generation.
Selection is done in F3. Seeds are removed from the selected plants.
These seeds are sown and plants raised. Crossing is again allowed in
these plants. This process is repeated for some generations till the
desired is obtained.
(c) Back cross method:
Fl offsprings when crossed with one of the parent is called backcross.
Backcross breeding method is used to transfer certain character such
as disease resistance, drought resistance etc. in the desired parent.
(d) Multiple cross method:
Cross between more than two parents is called multiple crossing. Its
objective is to transfer desirable characters from more than two
230
Plant Breeding
parents into one variety. Under this method first of all selected
pureline parents are made. Afterwards crossing is done between pairs.
The Fl are crossed. As shown in the diagram below the crosses are
made so as to combine all the desirable characters in the new variety:
AXB CXD EXF GXH
AB X CD EF X GH
ABCD X EFGH
ABCDEFGH
One of the advantages of this method is that desirable characters of
several parents can be transferred into one variety. New combinations
are also developed.
In this method there is a defect also. Because several parents are
involved in this method, therefore chances are to develop undesirable
combination of characters.
Hybridization in cross pollinated crops:
In cross pollinated crops viz. maize, rye, cucurbits, bajra, jute, clover,
fodder grasses, some fruit plants, etc. as breeding procedure
inbreeding ,is done to produce homozygous lines. The method is
described below: The desirable plants are selected. For inbreeding
the plant are self fertilize<! for about 9 - 10 generations. Then they
become homozygous (Table below) In inbreeding process some
abnormalities may occur ego Sterility, loss in size, chlorophyll
deficiency etc. After some generations the plant become homozygous
and then breed true. From the true breeding plants the desired ones
are selected. Then they are crossed. The offsprings develop heterosis.
Heterotic plants express the characters in vigorous forms.
Table: Increase in homozygosity in cross pollinated plant by selfing
for ten generation.
Generations selfed Homozygosity Heterozygosity
1 0 100
2 50 50
3 75 25
4 87.5 12.5
5 93.75 6.25
6 96.875 3.125
7 98.437 1.563
8 99.219 0.781
9 99.610 0.390
10 99.795 0.195
231
Following hybridization procedures are followed in cross pollinated

crops:
1. Single cross method: two good inbred lines are crossed. The
hybrid is used for cultivation.
2. Double cross method: In this method two single crosses are
made. For example A x Band C x D. Afterwards Fl of these
crosses are hybridized. Selection is made and the new variety is
produced. The variety NP 825 of wheat and Ganga 101 of maize
were developed by double cross method.
3 Triple cross method: In this method the Fl of two inbred lines
(Fl ofAx B) is crossed with a good variety. This way several
good genes are combined in one variety. Ganga 2 a good variety
of maize was developed by triple cross method.
Kinds of hybridization:
Depending on parental relationships hybridization can be of following
types:
1. Intervarietal cross
2. Interspecefic cross
3. Intergeneric cross
1. Intervarietal cross:
Desirable characters can be combined without difficulty. In wheat,
tobacco, cotion, tomato etc. by intervarietal crossings high yielding,
disease resistant, drought resistant, nutritive and high quality variety
can be developed. The variety NP 165 of wheat was developed by
crossing Australia Federation with NP 79.
2. Interspecific cross:
Hybridization between different species of a genus is called
interspecific cross. In several crops such as wheat, mustard, cotion,
luffa, tobacco etc. improvement have been done by this method.
Interspecific cross can be done in plants having one type of
chromosomes or different kinds of chromosomes. Some examples are
mentioned below:
Interspecific cross between Primula flOribunda (2n=18) and Primula
verticillata (2n=18) was made. The Fl chromosome number was
doubled. The tetraploid developed was named as Tetraploid Primula
kewensis (2n=36). Both the species have same number of chromosomes.
232
Plant Breeding
Example of interspecific crosses with different chromosome number

is Pennisetum typhoidium (2n=14) and Pennisetum purpureum (2n=28).
Pennisetum typhoidium suffers from rust disease whereas Pennisetum
purpureum is resistant to rust disease and has good quality of forage.
Its F1 hybrid chromosome number is doubled. Thus a hexaploid
drought resistant and high yielding new variety is produced.
Interspecific cross is usually recommended for vegetatively
propagated crops.
3. Intergeneric cross:
Crosses between different genus is called intergeneric cross. For crop
improvement inter generic crosses have been attempted in several
crops. For example sorghum and sugarcane; sugarcane and bamboo;
maize and teosenti; wheat and rye; mustard and radish etc. Success
of intergeneric cross is quite difficult. But if success comes results are
wonderful. In modern time intergeneric crosses are important. Some
intergeneric crosses are described below:
(1) Intergeneric cross between wheat(Triticum) and rye(Secale): The
hybrid obtained between Triticum and Secale is known as
Triticale or Secale triticum. It has a lot of economic importance.
(2) Intergeneric cross between a type of wheat(Triticum) and
Aegilops: Percibal (1935) reported a cross between Triticum
turgidum (2n=14) and Aegilops ovata(2n=14).He has produced a
useful hybrid of economic importance.
(3) Intergeneric cross between sugarcane (Saccharum)and
jowar(Sorghum). Venkatraman and Janki have crossed Saccharum
officinarum and Sorghum vulgare in Coimbatore, Tamil Nadu. The
obtained hybrid was early maturing and of economic importance.
Mutation breeding:
In modern times for improvement in crops, fruits and flowers
mutation breeding is widely attempted. For example in Coimbatore
(Tamil Nadu) on a variety GEB 24 of paddy by irradiation with X ray
a useful mutant variety 5782 was produced. This mutant variety is
dwarf, heavy tillering type and has other economic characters.
Similarly in Indian Agricultural Research Institute Jagdishan, Bhatia
and Swaminathan (1961) have treated the varieties NP 799 and NP
809 with X rays, phosphorus 32, & sulphur 35 and produced a mutant
variety NP 836. In the same way a good mutant variety of groundnut
was produced. In case of gram (Cicer arientinum) a mutant variety T
233
87 was evolved. Other chemicals ego ethyl methane sulphonate and

methyl ethane sulphonate have been used with success in several crops.
For further information on mutation please ,consult the chapter on
mutation.
Polyploidy breeding
Several crop varieties have been developed by induction of
polyploidy. To understand the role of polyploidy in plant breeding
please consult the chapter on polyploidy.
234
Index
Cartilage tissue engineering 210
A Caspase3 186
Accessory chromosome Zl cell cycle 157
Acquired Characters 13 Centromere 24
Adenocarcinomas 169 Chemotherapy 173
Adult stem cells 198 Chromomere 25
albinism 151 Chromonemata 24
Allele 13 Chromosomal Aberrations 107
allelomorph 13 Chromosomal Damage
Allopolyploid 80 Hypothesis 158
Alternate dominance theory 116 chromosomal mechanism 117
anamozygous mutation 72 chromosomal mutation 78
Anaphase 29 Chromosome 23
Aneuploidy 81 Chromosome Map 100
Annexin 186 chromosome theory 117
antigens 150 Cistron 136
Apoptosis 9,178 Colchicine treatment 84
Apoptosis-Inducing Factor 182 Comet Assay 186
Autopolyploid 80 Complete linkage 90
autosomes 77 Compound chromosomes 25
Contact first theory 97
B Cord blood stem cells 210
Cotton 223
Backcross 53 Crossing Over 94
Bacteriology 4 Cumulative genes 142
Basic genes l41 Cytokine therapy 177
Benign tumors 168 Cytokinesis 30
Biochemical mutation 71 Cytology 4
Biochemistry 4 Cytoplasmic Inheritance 112
Biological Therapy 175
Biopsy 170
Bone marrow 10
D
Bone marrow plasticity 206 death receptor signaling pathway 181
Bone marrow-derived stem Diandric sex linked inheritance 127
cells 204 Differentiation Gene Hypothesis 167
Bone tissue engineering 209 Digenic sex linked inheritance 126
Breakage first theory 97 dihybrid cross 45
DNA laddering 186
C Duplication 109
Cancer Genetics 155

Carcinomas 169
E
Cartilage 209 Ecology 4
235
ELlSA 186 Interferons 176

Embryology 3 Intergeneric cross 233
Embryonic Stem Cell 190 Interleukins 176
Embryonic Stem Cell Markers 191 Interphase 28
Eugenics 6,215 Interspecific cross 232
Euploid 79 Intervarietal cross 232
Extranuclear inheritance 113 Inversion 110
eye color 147
L
·F Lampbrush chromosomes 27
flower color 145 Law of dominance 38
Free Radical signaling pathway 182 Law of independent assortment 43
Free radical theory 75 Law of Segregation 41
Laws of Inheritance 37
Lethal gene 141
G Leukemias 169
Gene 131 Linkage 87
Gene Action 144 linkage strength 92
Gene Complex 141 lymphomas 169
gene mutation 72,78,137
gene therapy 5
Genetics 3
M
Genic balance theory 119 maize 88
Genomatic mutation 72 Marrow stromal cells 204
Germinal crossing over 94 Maternal effect 112
Germinal mutation 71 Matrix 24
Gynander 121 Meiosis 28, 30
Gynandromorph 121 mesenchymal stem cells 207
Metabolic differentiation theory 120
H Metaphase 28
metastatic tumors 168
Hematopoietic Stem Cell Markers191 mitochondrial pathway 181
Hematopoietic stem cells 204 Mitosis 28
Heterosis 67 Modifying gene 143
Holandric sex linked inheritance 127 Monoclonal antibodies 176
Hologenic sex linked 127 Monohybrid crosses 39
Hormone Therapy 175 Morphology 3
Horticultural crops 223 Multiple genes 142
Multipotent 187
Multipotent adult progenitor cells206
Mutagen 74
Immunologic Rejection 201 mutagen 138
Incomplete linkage 91 Mutation 71
Induced mutation 71 Mutation breeding 233
Intercallary deletion 108 Muton 136
Intercallary inversion 111
236
Sex limited character 131
N Sex Linked Inheritance 125
Neural Stem Cell Markers 192 Sex reversion 120
neurospora 151 Simple translocation 109
Nonspecific immunomodulating Somatic crossing over 94
agents 176 Somatic mutation7 1
Somatic Mutation Hypothesis 158
0 spermatogenesis
Spontaneous mutation
36
71
Oncogene Hypothesis 160 Spurious mutation 72
oogenesis 36 Statistics 4
Stem Cell 187
p Stem Cell Markers 191
Steriliza tion 219
Parasitic Infection Hypothesis 158 Strain theory 97
Phenylketonuria 151 Stromal Stem Cell Mi:1rkers 192
pigments in animals 146 Sugarcane 223
Plant Breeding 222 172
Surgery
Pleiotropic gene 142
Pluripotent 187
Polyhybrid cross 54 T
PolyplOidy 79 Target theory 75
Polyploidy breeding 234 Taxonomy 3
Potato 223 Telomerase Hypothesis 159
Prophase 28 Telomere 25
protein 149 Telophase 29
Proto-Oncogene Hypothesis 159 Terminal deletion 108
Psychology 4 Terminal inversion no
Pulse 223 Test Cross 52
Trans differentiation 199
R Trihybrid Cross 54
Tumor Suppressor Gene 163
Radiation Therapy 172
tumorgenesis 156
Receptor Tyrosine Kinases 159
TUNEL 185
Reciprocal translocation 109
Two-Stage Model Hypothesis 158
Recon 136
Resting stage 28
Rice 223 U
Unipotent 187
S
Salivery gland chromosome 25 V
Salpingectomy 219 Vesectomy 219
Sarcomas 169 Viral Infection Hypothesis 158
Secondary constriction 24
sex chromosome 76
Sex Determination 115 W
Sex influenced characters 130 Wheat 223
237

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GENETICS

International Book Distributing Co.

Dr. Debmalya Barh

International Book Distributing Co.

First Edition 2008

No part of this publication may be reproduced, stored in a retrieval

Composed & Designed at:

6 Mortohybrid cross between pea flowers. 40

4. Genetics and Cytology: Cytology of a living being is concerned

biological therapy, adjuvant therapy - combining two or more

characters of parents reach to their offsprings. This theory became

longitudinal structures to which he has given the name chromosome.

There are 3 different mechanisms of apoptosis: I-death signals

breeding of medicinal & aromatic crops the objective is to increase

Acclimatization: Ability of organism to adjusts with changed

Breeding: It is the science to change the genetic structure of plant or

Deficiency: Removal or absence of segments or gene from

Intersex: Intersex is the organism showing features of male and female.

is reduced to half .By meiosis cell division gametes are formed.

parthenogenesis: Developm~nt of organism without fertilization of

sex cells is parthenogenesis.

Recurrent selection: It is a method of plant breeding. Selection is

chromosome or other chromosome is known as translocation. If it

Fig.1: Structure of a typical chromosome.

Every chromosome has a kinetochore or centromere which is slightly

(8) Telocentric (b) Acrocentric (c) Metacentric (d) Sub metacentric

Fig.2: Shape of chromosome depending on centromeric position.

Fig. 3 b: Salivary gland chromosomes of fruit fly (Drosophila

(d) MitotIC metaphaSe

Fig.4: Diagramatic representation of various stage of mitosis.

6. Cytokinesis (Division of cytoplasm): Cytokinesis starts fonn

a. Leptotene or Leptonema: Cell size increases, chromosomes

to movement in centromeres the homologous chromosomes

Fig. 5: Diagrammatic representation of various stages of meiosis.

Differences in mitosis and meiosis are given below:

processes occurs only in their breeding season.

Material and method of John Gregor Mendel:

After proving the law of dominance in monohybrid crosses Gregor

Fig.6: Monohybrid cross between Fig. 7: Monohybrid cross between

Because black is dominant therefore B express the black colour though

Fig.8: Incomplete dominance in 4 0'dock plant <Mirabilis jalapa)

Fig.9: Hybridization between pink and white flower pea showing,the

Law of independent assortment:

Fig.IO: Process of assortment of chromatids during gamete formation.

1. Dominant gene is written in capitalletter. Wherever itis presented

RRYY (round & yellow)

Exceptions to the Law of independent assortment:

Red flowered pea =9

9AB 3Ab 3aB lab Reddish

As mentioned in the cross in case of brown glumes there is a gene B

9AB 3Ab 3aB lab

Hence, Phenotypic ratio = 9:6:1

9AB 3Ab 3aB lab

Gregor Mendel has explained a method - known as test cross. Its

of heterozygous recessive with homozygous recessive parent (test

Bull of either breed