Bio Lab Class 12
Bio Lab Class 12
Bio Lab Class 12
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Biology
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Class XII
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FOREWORD
The National Council of Educational Research and Training (NCERT) is the
apex body concerning all aspects of refinement of School Education. It has
recently developed textual material in Biology for Higher Secondary stage
which is based on the National Curriculum Framework (NCF)–2005. The NCF
recommends that children’s experience in school education must be linked
to the life outside school so that learning experience is joyful and fills the gap
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between the experience at home and in community. It recommends to diffuse
the sharp boundaries between different subjects and discourages rote learning.
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The recent development of syllabi and textual material is an attempt to
implement this basic idea. The present Laboratory Manual will be
complementary to the textbook of Biology for Class XII. It is in continuation
to the NCERT’s efforts to improve upon comprehension of concepts and
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practical skills among students. The purpose of this manual is not only to
convey the approach and philosophy of the practical course to students and
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teachers but to provide them appropriate guidance for carrying out
experiments in the laboratory. The manual is supposed to encourage children
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to reflect on their own learning and to pursue further activities and questions.
Of course the success of this effort also depends on the initiatives to be taken
by the principals and teachers to encourage children to carry out experiments
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evaluation will determine how effective this practical book will prove
to make the children’s life at school a happy experience, rather than
a source of stress and boredom. The practical book attempts to provide
space to opportunities for contemplation and wondering, discussion
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YASH PAL
Professor and Chairperson
National Steering Committee
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instructions. Today the stress is laid on the enquiry approach and
discussion method instead of lecture method of teaching. Biology is now
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something more than observation of living organisms. Study of Biology
includes microscopic observations to reveal minute internal details of the
organism, biochemical testing to understand complex reactions taking
place inside the organisms, experiments with live organism to understand
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various physiological processes and even much more. In other words
experiments in Biology truly represents an interdisciplinary approach of
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learning.
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The new syllabus of Biology has been designed to cater to the needs of
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pupil who are desirous of pursuing science further. The fundamental
objective of this course is to develop scientific attitude and desired
laboratory skills required for pursuing Biology as a discipline at this level.
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A similar approach has been taken while formulating the practical syllabus
of Biology for higher secondary stage. The practical syllabus includes
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and the questions are given in the book. The methodology of preparation
of any reagent, if required, has been given alongwith the requirements,
for the convenience of students and teachers. The questions are aimed to
develop learner’s understanding of the related problems. However, teacher
may provide help in case the problem is found to be beyond the capability
of the learner. Precautions must be well understood by the learners before
proceeding with the experiments and projects. In addition to the core
experiments enlisted in the syllabus for Class XII emphasis has also been
given for pursuing Investigation Project Work. It is expected that these
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to the participating teachers and subject experts who participated in the
review workshop and provided their comments and suggestions which helped
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in the refinement of this manual. We warmly welcome comments and
suggestions from our readers for further improvement of this manual.
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H UKUM SINGH
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Professor and Head
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Department of Education in
Science and Mathematics
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LABORATORY MANUAL DEVELOPMENT TEAM
MEMBERS
Animesh K. Mohapatra, Associate Professor, Regional Institute of Education,
NCERT, Ajmer
B.K. Tripathi, Professor, DESM, NCERT, New Delhi
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C.V. Shimray, Assistant Professor, DESM, NCERT, New Delhi
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N.V.S.R.K. Prasad, Associate Professor in Botany, Sri Venkateshwara College,
New Delhi
P.K. Durani, Professor (Retired), DESM, NCERT, New Delhi
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Sunita L. Varte, Assistant Professor, DESM, NCERT, New Delhi
S.P. Sinha, Professor of Zoology (Retired), TM Bhagalpur University, Bhagalpur
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V.V. Anand, Associate Professor, Regional Institute of Education, Mysore
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MEMBER-COORDINATOR
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Professor, Department of Zoology, University of Delhi, Delhi; K.K. Sharma, Professor Department
of Zoology, M.D.S. University, Ajmer; M.M. Chaturvedi, Professor Department of Zoology,
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University of Delhi, Delhi; Nazir Ahmad Kakpori, Department of Education, Govt of Jammu &
Kashmir, Srinagar; Reena Mohapatra, St. Stephen’s Senior Secondary School, Ajmer; Savita
Sharma, Mount Carmel School, Dwarka, New Delhi; Savithri Singh, Professor and Principal,
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Acharya Narendra Dev College, New Delhi; Shalu Dhawan, Amity International School, Saket,
New Delhi; Shivani Goswami, Mother’s International School, New Delhi; V.K. Srivastava, Reader
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in Zoology, J.N. College, Pasighat; Vijay Kumar, Delhi State Science Teacher Forum, New Delhi.
We also acknowledge the contributions of Anil Kumar and Binita Kumari, Junior Project
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Fellows, DESM, NCERT, New Delhi.
Special thanks are also due to Hukum Singh, Professor and Head, DESM, NCERT for his
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Rajput and Girish Goyal, DTP Operators, for helping in shaping this laboratory manual. The
contributions of Publication Department of NCERT in printing out this laboratory manual are
also duly acknowledged.
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CONTENTS
FOREWORD iii
PREFACE v
Introduction 1
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Exercise 1 : To study the reproductive parts of commonly 5
available flowers
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Exercise 3 : To study pollen tube growth on stigma 14
Exercise 4 :
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To study the discrete stages of gametogenesis in 16
mammalian testis and ovary
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Exercise 17 : To study the ecological adaptations in plants living 62
in xeric and hydric conditions
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Exercise 18 : To study the adaptations in animals living in xeric 65
and hydric conditions
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Exercise 19 : To determine the pH of different water and soil 68
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samples
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Exercise 20 : To study turbidity of water samples 71
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Introduction
Laboratory is a place where ideas and concepts can be tested through
experiments. Biology, like any other discipline of science, is based on
experimental work and therefore practical forms an integral part of learning.
Biology laboratory provides a unique learning environment where learners
inculcate scientific temper, develop relevant skills and get exposed to realms
of techniques and methodologies of scientific investigations. Laboratory
investigations in Biology increase the reasoning abilities, bring scientific
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attitude in a learner and also help in acquisition of skills of scientific processes.
Also, observation of nature and the living organisms found in it is no less
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important for the understanding of many aspects of the subject especially
the diversity of the living organisms, their systematic study, their relationships
among themselves and with the environment. Knowledge in the field of
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Biology can be acquired or constructed only on the basis of correct
observations and experimentally verifiable processes.
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Biology laboratory thus provides the learners an environment where the
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process of learning is facilitated by hands-on experiments. Biology is a
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unique discipline in the sense that it does not merely deal with the study of
morphology, anatomy, physiology and reproduction of the living organisms,
rather, understanding of the subject requires understanding of a number of
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habit, habitat and external features of the plants or animals, or, it may involve
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certain manipulations like dissection and section cutting of the parts of the
organisms to study the minute details. Very often observation and study of
the magnified image of the minute parts under a microscope provides a
better insight about the features of the organisms. However, microscopic
study involves certain specific skills depending on type of the organisms/
tissues/cells to be studied. It involves specific preparations (peeling, section
L ABORATORY MANUAL: BIOLOGY
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In order to perform experiments successfully, a learner needs to go to
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the Biology laboratory well prepared. This includes the following:
1. Laboratory Record Book: For maintaining all the information including
recording of data and its interpretation.
2.
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Dissection Box: A dissection box is required in the Biology laboratory
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for various purposes like handling and manipulation of living materials,
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performing experiments, preparation of slide, etc. A dissection box
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should contain scissors (two pairs, one small with fine tip and one
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larger), scalpels (one small and one medium sized), forceps (two, one
small with sharp fine tips and the other medium sized with blunt tips),
dissecting needles (two), razor, hand lens, dropper, fine brush, etc.
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3. Laboratory Manual
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in which the aim of the experiment, its principle, list of the materials to be
used, procedure, observation table (if required), inference and discussion
should be given. Necessary precautions to be taken should also be
mentioned appropriately in the procedure or at the end. There are a few
experiments in which field visit is essentially required. For this all the
necessary preparations (materials, equipments, reagents and chemicals)
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IENTRODUCTION
XERCISE 1
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• Appropriately label your drawing. Parts of the drawing should be
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indicated by straight horizontal line or arrow. Two lines or arrow should
never cross each other. As far as possible, labelling should be done on
the right side of the drawing. An appropriate legend or heading of the
drawing should also be given below it.
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About the Manual
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The main objective of the manual is to introduce the students of higher
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secondary stage to the fascinating world of plants, animals and microbes
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and their complex biological phenomena. The manual covers a complete
description of the experiments and exercises. The suggested experiments
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cover almost all the units/topics including those on diversity in living world,
plant, animal and human physiology, genetics, bio-technology and human
welfare and environment. A standard format has been used to describe each
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L ABORATORY MANUAL: BIOLOGY
• Observation and Results: This section deals with the recording of all
observations made during the experiment. Students are advised to
consider the entire data. Data can be represented in the form of tables,
graphs and histograms wherever possible. Use of units in which various
quantities are measured has been indicated in the manual.
• Discussion: Included in this heading is a statement of the conclusions
drawn from the experimental results and compared thesis (wherever
possible) with any comparable data from other sources. The relevance
of the conclusions drawn from the experimental results to the various
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processes under investigation and to the life of plant, animal and microbes
has been prompted out.
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• Precaution: This section contains all the necessary precautions to be
taken during experimentation to obtain results free of errors. However,
attempts have been made to mention required precautions along with
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the procedure also.
A great emphasis has been laid on a student getting valid results and
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interpreting them. It is essential that the teacher should properly explain
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each experiment so that inexperienced students will be able to obtain accurate
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results within a reasonable time. Teachers are also expected to help students
in identifying errors and mistakes committed during experiments and ways
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for correcting them. It is possible that some of the students may undoubtedly
be capable of doing more sophisticated work than that represented in the
manual. But introductory course of this sort has been designed to help all
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Exercise 1
Aim: To study the reproductive parts of commonly available flowers
Principle: The male reproductive parts of a flower are the stamens collectively called
androecium and the female reproductive parts are the carpels/pistils collectively called
gynoecium. The individual units of stamen consist of a filament, which supports the anther
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lobes. Gynoecium consists of stigma, style and ovary. Many variations are found in different
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characteristics of both the stamens and carpels. We shall try to study these variations in the
reproductive parts of flowers in the exercise.
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Requirement: Commonly available flowers, needles, forceps, razor/scalpel blade, brush, slides,
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cover slip, watch glass, magnifying lens, dissecting microscope, compound microscope, etc.
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Procedure
(i) Familiarise with the terms to describe the reproductive parts of flowers
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(ii) Observe the flower with the naked eye, hand lens or under a dissecting
microscope. Study their reproductive parts and count the number of
stamens and record their cohesive and adhesive features.
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(iii) Cut L.S. of the flower and place it on a slide to observe the following
characters:
(a) Placement of anthers
(b) Position of the ovary: epigynous/perigynous/hypogynous.
(iv) Mount one stamen on a slide and study the following characters:
(a) Attachment of filament to anther
(b) Dehiscence pattern of the anther lobes for discharge of pollen.
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(v) Cut T.S. of anther lobe to observe the number of pollen sacs.
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(vi) Mount the pistil on a slide and study style, stigma and ovary. Record
the number of stigma and nature of pistil.
(vii) Cut T.S. of ovary, mount it on a slide and observe
(a) Number of locules in the ovary
L ABORATORY MANUAL: BIOLOGY
Questions
1. Name the most common type of placentation observed.
2. What is the most common type of dehisence pattern in anthers?
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3. Name a few unisexual flower-bearing plants studied by you.
4. “Flower is a modified shoot.” Justify the statement based on your observation.
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EXERCISE 1
Annexure 1
Androecium
Number of stamens The number of stamens may vary from a few to many in dif-
ferent flowers
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(Fig. 1.1 a–e) following type:
(i) Syngenesious : Filaments free and anthers united,
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e.g., Sunflower.
(ii) Synandrous : Stamens fused all through their length,
e.g., Cucurbita
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Adelphous condition can be
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(a) Monoadelphous—United to form 1 bundle, e.g.,
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China rose
(b) Diadelphous—United to form 2 bundles, e.g., Pea
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(c) Polyadelphous—United into more than two
bundles, e.g., Lemon
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family
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Dehiscence pattern (i) Porous : Pollens released through pores, e.g., Brinjal,
(Fig. 1.4 a,b) Potato
(ii) Longitudinal: Pollens released through the longitudinal
slit of another lobes, e.g., China rose, Cotton
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L ABORATORY MANUAL: BIOLOGY
Gynoecium
Number of stamens The number of stamens may vary from a few to many in
different flowers
Position of ovary (i) Epigynous: Position of ovary inferior to other floral parts,
(Fig. 1.5 a–d) e.g., Mustard, China rose
(ii) Perigynous : Other floral parts are attached around the
ovary, e.g., Apple, Guava
(iii) Hypogynous: Position of ovary superior to other floral
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parts, e.g., Sunflower
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Cohesion If number of carpels is more than one, they may be
(Fig. 1.6 a–c) (i) Apocarpous : Carpels are free. Each carpel has its own
style and stigma, e.g., Rose
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(ii)
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Number of locules in ovary Vary from one to many
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(i) Unilocular : One locule, e.g., Rose, Pea
(ii) Bilocular: Two locules, e.g., Datura
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(iii) Multilocular : Many locules, e.g., Lady’s finger, China rose
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Placentation (i) Marginal : The placenta forms a ridge along the ventral
(Fig. 1.7 a–e) suture of the ovary and the ovules are borne on this ridge,
e.g., Pea
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EXERCISE 1
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(d)
(e)
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Fig.1.1 Cohesion of stamens: (a) Syngenesious (b) Synandrous
(c) Monoadelphous (d) diadelphous (e) Polyadelphous
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L ABORATORY MANUAL: BIOLOGY
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(a) (b)
Fig.1.4 Dehiscence pattern of anther: (a) Porous (b) Longitudinal
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EXERCISE 1
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(a) (b) (c)
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(d) (e)
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Fig.1.7 Placentation: (a) Marginal (b) Axile (c) Parietal (d) Free central (e) Basal
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Exercise 2
Aim: To calculate percentage of pollen germination
Principle: In nature, pollen grains germinate on the compatible stigmas of the carpel. Pollen
grains can also be induced to germinate in a synthetic medium. During germination, intine
(inner wall) of pollen grain emerges out as pollen tube through one of the germ pores in exine
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(outer wall).
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Requirement: Calcium nitrate, boric acid, sucrose, distilled water, petridish, slides, coverslips,
brush, needle, microscope, and mature pollen grains of Tradescantia/balsam/Jasmine/lily/
pomegranate/grass/Vinca/China rose/Petunia.
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Procedure
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(i) Prepare the pollen germination medium by dissolving 10g sucrose,
30mg calcium nitrate and 10mg boric acid in 100ml of distilled water.
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(v) Count (a) total number of pollen grains seen in the microscope field,
and (b) the number of pollen grains that have germinated.
Observation
Several pollen grains germinate and put forth pollen tubes. Count the total
number of pollen grains and the number of germinated pollen grains in 3-5
different microscope fields. Tabulate your observations and calculate the
percentage of pollen germination.
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Nameoftheplantusedassourceofpollen……………………………
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Number of Total number of pollen (N) Total number of pollen germinated (n) % pollen germination
observation n
× 100
N
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2.
3.
4.
5.
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Average
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Discussion
Although pollen grains of many species germinate in this medium, the
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percentage of germinations and the time taken for germination varies in
different species. Draw a germinating pollen grain and label.
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Questions
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Exercise 3
Aim: To study pollen tube growth on stigma
Principle: Pollen grains germinate and form pollen tubes after they get deposited by the process
of pollination on compatible stigma. Pollen tube, made up of cellulose, is an extension of the
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inner wall of pollen grain (intine). It emerges through one of the germ pore and passes through
tissues of stigma and style to reach the ovule. The growing pollen tube is observed by staining
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with cotton blue.
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(Lady's finger), beaker, water, slides, cover slips, cotton blue stain, microscope, brush, needle.
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Procedure
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Observation
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traversing through the tissues of stigma and Fig.3.1 Growth of pollen tube
style (Fig. 3.1). in the style of a carpel
EXERCISE 3
Discussion
Pollen tubes are seen amidst the stylar tissue. Many pollen tubes may be
seen. Trace the origin of pollen tubes to the pollen grains present around
the surface of the stigma.
Questions
1. Can pollen grains of one plant species germinate on stigma of other species?
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Give reasons.
2. Do all pollen tubes reach the ovules?
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3. Are all the pollen tubes of equal length? If not, why?
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Exercise 4
Aim: To study the discrete stages of gametogenesis in mammalian testis and ovary
Principle: In all male and female organisms gamete formation takes place in their gonads, i.e.,
testis and ovary respectively. The process of gamete formation, called gametogenesis involves
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meiotic cell division. The gametogenic development in testis is called spermatogenesis and in
ovary it is oogenesis. They exhibit marked differences and can be examined in transverse section
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(T.S.) of these organs.
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lens-cleaning paper and cleaning fluid
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Procedure
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(i) Clean the slide and microscope’s eye and objective lenses with the
help of lens cleaning paper using any cleaning fluid.
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(ii) Place the slide on the stage of the microscope and observe first under
lower magnification and then in higher magnification. Observe various
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Observation
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T.S. of testis
(i) You will observe a large number
of seminiferous tubules under
Seminiferous tubule
lower magnification. Observe a
complete tubule in higher
magnification and view various
stages of gamete development Spermatozoa
from periphery towards lumen
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(ii) In T.S. of testis the space between tubules are filled with blood vessels
and a specific cell type called Leydig's cell or Interstitial cells.
T.S. of Ovary
(i) In the section of ovary, there is a mass Graafian Follicle
of tissue lined with germinal
epithelium. Inside that you will Antrum
observe an ovum, which is a cell
surrounded by one to several layers
of follicular cells. As the ovum
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matures, the number of surrounding
follicular cell layer increases (Fig. 4.2). Corpus luteum
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(ii) In the later stage of follicular Corpus albicans
development a cavity called antrum
appears.
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(iii) The cavity gets further enlarged and
the follicle grows bigger. This is the
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stage of Graafian follicle ready to release the ovum (ovulation).
(iv) In the next stage, you may notice a Corpus luteum, and/or Corpus
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albicans, which differ from each other and also from Graafian follicle in
their features.
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Discussion
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observed.
Questions
1. What would happen if meiosis fails to occur in gametocyte?
2. At which stage of follicular development, is ovum released?
3. Spermatogenesis is a continuous process. Justify the statement.
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Exercise 5
Aim: To study and identify various stages of female gametophyte development in the ovary
of a flower
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situated deep inside the ovule. An ovule generally has one female gametophyte. Development
of female gametophyte begins with megaspore mother cell. Most common type of female
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gametophyte is the monosporic, 8-nucleate, 7-celled type.
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stages of female gametophyte development and microscope
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Procedure
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(i) In a V.S. of ovary we generally find several ovules. Carefully observe
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Observation Chalaza
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Egg
Synergids
Central cell
Secondary nucleus
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(a) (b) (c) (d) (e) Antipodals
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(b) 4-nucleate stage (c) 8- nucleate stage (d) 8- nucleate stage
showing 3+2+3 distribution of nuclei (e) mature embryo sac.
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(iii) Note the contents of embryo sac, namely, an egg apparatus
(2 synergids and egg) at micropylar end, secondary nucleus in the
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center and three antipodal cells at the chalazal end (Fig. 5.2).
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Questions
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Exercise 6
Aim: Preparation and study of mitosis in onion root tips
Principle: Somatic growth in plants and animals takes place by the increase in the number of
cells. A cell divides mitotically to form two daughter cells wherein the number of chromosomes
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remains the same (i.e., unchanged) as in the mother cell. In plants, such divisions rapidly take
place in meristematic tissues of root and shoot apices, where the stages of mitosis can be
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easily observed. In animals, mitotically dividing cells can be easily viewed in the bone marrow
tissue of a vertebrate, epithelial cells from gills in fishes and the tail of growing tadpole larvae
of frog.
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Requirement: Onion bulbs, wide mouth glass tubes/jar/bottle, glacial acetic acid, ethanol
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2-4% acetocarmine/acetoorcein stain, N/10 HCl, spirit lamp/hot plate, slide, cover slips,
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blotting paper, molten wax/nail polish and compound microscope
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Procedure
Growing of root tips
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Select a few medium-sized onion bulbs. Carefully remove the dry roots
present. Grow root tips by placing the bulbs on glass tubes (of about 3–4
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cm. diameter) filled with water. Care should be taken so that the stem portion
of the bulb (basal part) just touches the water. A few drops of water may be
added periodically to compensate evaporation losses. New roots may take
3–6 days to grow. Cut 2–3 cm long freshly grown roots and transfer them to
freshly prepared fixative, i.e., aceto-alcohol (1:3:: glacial acetic acid : ethanol).
Keep the root tips in the fixative for 24 hours and then transfer them to 70%
ethanol (for preservation and use in future). Onion root-tip cells have a cell
cycle of approximately 24-hour duration, i.e., they divide once in 24 hours,
and this division usually takes place about two hours after sunrise. Therefore,
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roots grown on water should be cut only at that time to score maximum
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Preparation of slide
Take one or two preserved roots, wash them in water on a clean and grease-
free slide. Place one drop of N/10 HCl on the root tip followed by 2–3 drops
of aceto-carmine or aceto-orcein stain on it. Leave the slide for 5–10 minutes
EXERCISE 6
on a hot plate (or warm it slightly on spirit lamp). Care should be taken that
the stain is not dried up. Carefully blot the excess stain using blotting paper.
Now cut the comparatively more stained (2–3 mm) tip portion of the root
and retain it on the slide and discard the remaining portion. After
(10–20 seconds) put one or two drops of water and blot them carefully using
blotting paper. Again put a drop of water on the root tip and mount a cover
slip on it avoiding air bubbles. Place the slide in between the folds of blotting
paper using the fingers in such a way that the cover slip mounted on the
slide is properly held. Now slowly tap the cover slip using the blunt end of a
pencil so that the meristematic tissue of the root tip below the cover slip is
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properly squashed and spread as a thin layer of cells. Carefully seal the
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margins of the cover slip using molten paraffin wax or nail polish. This
preparation of onion root tips cells is now ready for the study of mitosis.
Study of slide
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Place the slide on the stage of a good quality compound microscope. First
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observe it under the lower magnification (10 X objective) to search for the
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area having a few dividing cells. Examine the dividing cells under higher
magnification of the microscope to observe the detailed features of mitosis.
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Observation
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The stages of mitosis can be broadly categorised into two parts: karyokinesis
(division of nucleus) followed by cytokinesis (division of cytoplasm, and
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ultimately of the cell). Those cells, which are not in the phases of cell division
are considered to be in interphase. You may observe that most of the cells
in a microscope field are in interphase
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Interphase
The cells are mostly rectangular, oval or even circular in shape, with almost
centrally situated densely stained nucleus. The chromatic (coloured) material
of the nucleus is homogeneous and looks granular. The boundary of the
nucleus is distinct. One or few nucleoli (sing: nucleolus) can also be observed
inside the nucleus (Fig. 6.1a).
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Stages of Mitosis
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(a) Prophase
Intact nuclear outline is seen. The chromatin (seen as a homogeneous
material in the nucleus at interphase) appears as a network of fine threads
(chromosomes). Nucleoli may or may not be visible (Fig. 6.1b).
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L ABORATORY MANUAL: BIOLOGY
a. Interphase
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b. Prophase
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c. Metaphase
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d. Anaphase
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e. Telophase
(b) Metaphase
The nuclear membrane disappears. Chromosomes are thick and are seen
arranged at the equatorial plane of the cell (Fig. 6.1c). Each chromosome at
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EXERCISE 6
this stage has two chromatids joined together at the centromere, which can
be seen by changing the resolution of the microscope. Nucleolus is not
observed during metaphase.
(c) Anaphase
This stage shows the separation of the chromatids of each chromosome. The
chromatids separate due to the splitting of the centromere. Each chromatid
now represents a separate chromosome as it has its own centromere. The
chromosomes are found as if they have moved towards the two poles of the
cell. The chromosomes at this stage may look like the shape of alphabets 'V',
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'J' or 'I' depending upon the position of centromere in them. Different anaphase
cells show different stages of movement of chromosomes to opposite poles,
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and they are designated to represent early, mid and late anaphase (Fig. 6.1d).
(d) Telophase
Chromosomes reach the opposite poles, lose their individuality, and look
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is
like a mass of chromatin (Fig. 6.1e). Nuclear membrane appears to form the
nuclei of the two future daughter cells.
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Cytokinesis
bl
E
In plants, a cell plate is formed in the middle after telophase. The plate can
be C
be seen to extend outwards to ultimately reach the margin of the cell and
divide the cell into two. Such cell plates are characteristic of plant cells
(Fig. 6.2). However, in an animal cell, the two sides of the cell show inpushings
to N
or constrictions formed from the peripheral region in the middle of the cell,
which grow inward and meet to divide the cell into two daughter cells.
Draw labelled diagrams of all the phases of mitosis.
©
Discussion
no
Mitotic index (MI) is defined as a ratio of the total number of dividing cells (n)
and the total number of cells (N) in a particular focus chosen randomly under
n
the microscope and is calculated as MI = N × 100 . By randomly selecting
5 to 10 such foci, one can estimate the mitotic index for a given type.
23
L ABORATORY MANUAL: BIOLOGY
d
Prophase Metaphase Anaphase Telophase
he
1. Cell morphology
2. Nuclear morphology
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3. Chromosomes/chromatids
is
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Questions
bl
E
1. Suggest names of a few tissues, which are suitable for the study of mitosis.
be C
24
Exercise 7
Aim: Study of stages of meiosis using permanent slides
Principle: Meiosis is a type of cell division in which the number of chromosomes is halved
(from diploid to haploid) in the daughter cells, i.e., the gametes. The division is completed in
two phases, meiosis I and meiosis II. Meiosis I is a reductional division in which the chromosomes
d
of homologous pairs separate from each other. Meiosis II is equational division resulting in the
he
formation of four daughter cells. Stages of meiosis can be observed in a cytological preparation
of the cells of testis tubules or in the pollen mother cells of the anthers of flower buds.
pu T
Requirement: Permanent slides of meiosis and compound microscope
is
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Procedure
bl
E
Place the slide on the stage of the microscope and search for the dividing
cells using lower magnification. When dividing cells are located observe them
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Observation
Observe various stages of meiosis and identify them on the basis of the specific
features given in the table 7.1. A significant number of cells will be in the
©
identified in it.
no
d
(a) Leptotene
he
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bl (b) Zygotene
E
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to N
©
(c) Pachytene
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no
(d) Diplotene-Diakinesis
26
EXERCISE 7
d
(d) Diplotene (diplos = double)
The homologous chromosomes (each made up of two chromatids)
he
show distinct separation from each other except at few regions where
attachments are seen (Fig. 7.1d). These are chiasmata (sing.
chiasma) representing the site of exchange of the parts between
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two homologous chromosomes (i.e. crossing over).
is
(e) Diakinesis (Dia = opposite; kinesis= separation or movement)
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(i) The homologous pair of chromosomes appear more shortened,
bl
thick and prominent (Fig. 7.1d).
E
(ii) Chiasmata can be still observed.
(iii) All the homologous pairs appear scattered in the cell.
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2. Metaphase I Homologous chromosomes are still in pairs, and are arranged along
the equatorial plane of the cell (Fig. 7.2a). At this stage, the number
of bivalents can be counted. Chiasmata may still be seen in a few
to N
bivalents.
3. Anaphase I The chromosome pairs appear to have moved towards the two
opposite poles of the cell. At the later stage, the anaphase - I may
©
Meiosis II
1. Prophase II (i) Distinct thread- like chromatin fibres or rod- shaped chromosome
are seen.
27
L ABORATORY MANUAL: BIOLOGY
(b) Anaphase I
(a) Metaphase I
d
he
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(c) Telophase I
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bl
Fig. 7.2 Phases of Meosis I (a-c) – actual microscopic view on left side
and its diagrammatic representation on the right hand side.
E
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to N
©
(a) Metaphase II
t
no
(b) Anaphase II
28
EXERCISE 7
d
forming tetrad.
(iii) In the metaphase I of meiosis, a few chiasmata are observed,
he
where as no chiasmata are observed during metaphase II.
3. Anaphase II The two chromatids of each chromosome after separation appear
to lie at the two poles of the cell (Fig. 7.3b).
is
meiotic division on the basis of chromatids: In anaphase I, each
chromosome has two distinct chromatids, but in anaphase II, each
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chromosome is represented by one chromatid only.
4. Telophase II The separated chromosomes appear decondensed and form nuclei
E
(Fig. 7.3c).
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to N
Questions
1. What is the significance of meiosis?
2. What is synapsis and crossing over?
©
29
Exercise 8
Aim: To study the blastula stage of embryonic development in mammals, with the help of
permanent slide, chart, model or photograph
Principle: The zygote undergoes a few cycles of mitotic divisions to form a solid ball of cells called
d
morula. The cells continue to divide and at a later stage a cavity is formed within it. This stage is
blastula. The internal structural details of blastula can be observed in its transverse section.
he
Requirement: Permanent slide, chart/model of T.S. of blastula, compound microscope,
lens cleaning fluid and paper
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is
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Procedure
E
Observe the slide under lower magnification of the microscope. In case of
chart/models/photographs, note the feature of blastula in your practical
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Observation
to N
Questions
1. What are the differences between blastula and morula?
2. What are the main structures you observe in T.S. blastula?
3. Match the stages in column I with features in column II
Column I Column II
(a) Trophoblast (i) Dividing cells of the morula
(b) Morula (ii) Outer layer of blastula
d
(c) Blastocoel (iii) Solid ball of cells
(iv) Cavity
he
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bl
E
be C
t to N
©
no
31
Exercise 9
Aim: To verify Mendel's Law of Segregation
Principle: When two pure lines with contrasting forms of a particular character (phenotypes) are
crossed to produce the next generation (F1 generation), all the members of the progeny are of only
d
one phenotype i.e. of one of the two parents. The phenotype that appears is called dominant, and
the one that does not appear is called recessive. When the F1 plants are selfed, the progeny i.e. the
he
F2 generation is in the ratio of 3 dominant: 1 recessive (¾: ¼ or 75%: 25%). This reappearance of
the recessive phenotype in F2 generation verifies law of segregation.
pu T
Requirement: 64 yellow and 64 green plastic beads, all of exactly same shape and size, (when
is
beads are not available, pea seeds may be coloured using paint, these beads represent the
re R
gametes of a specific trait), plastic beakers/petri dishes and a napkin/hand towel
bl
E
Procedure
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(ii) Take a bead from each container and place them together (it represents
fertilisation) on the napkin spread before you on the table. (One student
to take out beads and to put in the hands of the other student who will
put them on the table).
(iii) Just like the previous step, continue to pick beads and arrange them
in pairs. Thus 64 pairs of beads are obtained representing the 64
heterozygous F1 progeny.
Note that all the F1 individuals are represented by one yellow and one
t
green bead.
no
(vi) To obtain the F2 generation, one student would withdraw one bead
from one beaker labelled male and one from the other beaker labelled
female keeping his/her eyes closed (to ensure randomness), and put
them together in the stretched palm of the partner, who will put them
together on the napkin spread over the table. Continue this process till
all the beads are paired. Thus 64 offsprings of F2 are obtained.
(vii) Note the genotype (YY or Yy or yy) of each pair, and their possible
phenotype.
(viii) Have six repeats of the experiment (steps i to vii) with partners changing
d
their roles. Pool all the data from the six repeats together.
he
(ix) Calculate the genotypic and phenotypic ratios of your pooled data.
Note that larger the sample size, more accurate is the result.
Observation
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Record the result in the following table:
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bl
Generation Repeat No. Total no. of Genotype (s) Phenotype (s)
individuals YY Yy yy
E
F1 1.
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2.
3.
to N
4.
5.
6.
©
Total
F2 1.
2.
3.
4.
5.
6.
Total
t
no
33
L ABORATORY MANUAL: BIOLOGY
Discussion
The results are so because each diploid individual contains two copies of
every gene - one copy on each of the two homologous chromosomes. These
two copies of the gene may be of similar type (YY or yy) or are dissimilar Yy.
The former (YY or yy) are called homozygous for that particular character,
and the Yy are called heterozygous ones. The pure lines in the above cross
are homozygous ones, which contributed only one copy of their gene (as a
result of meiosis) to their F1 progeny to restore its diploid nature with genotype
Yy (heterozygous) where only one form (allele) is expressed (dominant) and
d
the other form (allele) is not expressed (recessive). This is the phenomenon
of Dominance.
he
When the F1 individuals are crossed together to raise the F2 generation,
each F1 individual produces two types of gametes: 50% having dominant
allele, and the remaining 50% having recessive allele. These gametes undergo
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is
random fusion during fertilisation to produce the F2 generation. According
to simple probability of mixing of opposite sex gametes (sperms and ova),
re R
bl
offsprings of three genotypes are likely to appear as follows: [(half of gametes
of Y type + half of remaining gamete y type) X (half gametes of Y type + half
E
of remaining gamete of y type)] = One-fourth of F2 individuals of YY phenotype
+ half of F2 individual Yy type + one-fourth of F2 individul of yy type. Among
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from the influence of the dominant allele (Y). This is called Law of Segregation
of the alleles.
Questions
1. Do you expect the same results in terms of 3:1 ratio in F2 if you had started with
smaller number of beads (say 10 beads)?
t
no
34
Exercise 10
Aim: To verify the Mendel’s Law of Independent Assortment
Principle: In a dihybrid cross, the segregation of one gene pair is independent of the segregation
of the other pair. It means that genes of two different traits assort independently to give a
probability ratio equal to segregration probability ratio of one allele pair X segregation probability
ratio of other allele pair, which comes to, (3:1) X (3:1) = 9:3:3:1
Requirement: Plastic beakers; 64 plastic beads each of yellow, green, red and white to represent,
yellow and green colour of seed coat and red and white flowers respectively and napkin/hand
towel
Procedure
Students are to work in pair.
The following steps are to be followed sequentially:
(i) Place 64 beads of each colour in four separate beakers.
(ii) Put the beakers containing the yellow and red beads on your left side,
and those containing the green and white beads on your right side.
The beakers on your left side represent plants bearing yellow seed and
red flower (dominant character YY, RR). Beakers on the right side
represent plants bearing green seeds and white flowers (recessive
character yy, rr). These are the two parental types having contrasting
forms of two different characters.
(iii) Stir the beads in each beaker with a pencil/pen. Each bead now
represents alleles in the male and female gametes.
(iv) Pick up one yellow, one green, one red and one white bead, and put
them together on the napkin spread on the table.
(v) Continue picking up and putting together of the beads of all colours
as mentioned in the previous step, till all the beads are utilised.
(vi) Note that in all, 64 such 4-bead clusters are obtained representing the
F1 individuals. Ascertain their genotype and phenotype.
(vii) Next step is to cross these F1 individuals to raise the F2 generation. Let
us suppose half of the 4-bead clusters (32 clusters) represent the male
parents and the remaining half (32 clusters) the female parents. Now
put the 32 red and 32 white beads together in one beaker (numbered-
I), and similarly put 32 yellow and 32 green beads together in other
L ABORATORY MANUAL: BIOLOGY
Female F1 Male F1
(viii) Stir the beads in each beaker with a pencil. In order to raise the F2
generation, pick up (with eyes closed) one bead from the beaker-I of
female and one bead from the beaker-III of the male, and put into the
palm of the partner student. Similarly, pick up one bead each from the
beaker-II of female and beaker IV of male to put in the palm of the
partner. This partner would now keep all the four beads together (to
represent the F2 individual). Continue this process till all beads are
utilised. At the end, 64 F2 individuals (each represented by a 4-bead
cluster) are obtained.
(ix) Determine the genotype and phenotype of each of the 64 F2 individuals
and write down the number of individuals of different genotypes and
phenotypes in the tabular form (given below), remembering that Y
(yellow seed colour) is dominant over y (green seed) and R (red flower)
is dominant over r (white flower).
(x) Repeat the whole procedure (steps i to ix) six times, and tabulate your
results.
Observation
Tabulate the results as follow:
Symbol (-) indicates the presence of corresponding dominant or recessive
allele e.g. Y or y and R or r.
Summarise your results (adding together the data of all the six repeats)
F1 Generation
(a) Total number of individuals: _________________________
(b) Phenotype (s) _________________________
(c) Genotype (s) _________________________
36
EXERCISE 10
F1
1.
2.
3.
4.
5.
6.
Total
F2
1.
2.
3.
4.
5.
6.
Total
F2 Generation
(a) Total number of individuals _________________________
(b) Phenotypes _________________________
(c) Number of individuals in each phenotypic class:
Number Phenotype
__________________ _____________________
__________________ _____________________
__________________ _____________________
__________________ _____________________
(d) Phenotypic ratio _____________________
(e) Genotypic ratio _____________________
37
L ABORATORY MANUAL: BIOLOGY
Discussion
The four phenotypic classes in the F2 generation are in ratio of 9:3:3:1 as
expected from the Law of Independent Assortment. The genotypic ratio
would be (1:2:4:2): (2:1):(2:1):1.
Note
1. In case six repeats of the experimental procedure are not feasible due
to time limitations, either the number of repeats be slashed down to
three or the data from single repeat of six different pair of students may
be pooled together to make the final calculations.
2. This Law of Independent Assortment was later found to be true only for
traits present on two different homologous pair of chromosomes, that
is, the two are not linked together. The linked traits do not assort
independently, rather they are inherited together (linked) except when
crossingover separates them.
3. It is quiet likely that you may not find your data exactly in the
expected ratio, instead almost approximate to it. The statistical
significance of this deviation from the exact expected ratio due to
probality can be checked using chi-square (χ 2) test, about which
you will study in higher classes.
Questions
1. Linked traits fail to assort independently. Explain.
2. How is independent assortment of alleles important from the point of view of
variation?
38
Exercise 11
Aim: Preparation and analysis of Pedigree Charts
Principle: The Mendelian concept of dominance and segregation can also be studied in humans
by preparing and then analysing the pedigree charts. The internationally approved symbols for
indicating males and females, marriages, various generations (I, II, III), etc., are given below.
Requirement: Information about characters/traits in a family for more than one generation
Procedure
Select a family in which any one of the monogenic traits such as tongue
rolling, widow's peak, blood groups’, red-green colour blindness, dimple in
L ABORATORY MANUAL: BIOLOGY
the cheek, hypertrichosis of ear, hitch-hiker's thumb, etc., is found. Ask the
person exhibiting the trait to tell in which of his/her parents, grand parents
(both maternal and paternal), their children and grand children the trait in
question is present. Among surviving individuals the trait may also be
examined. The information made available is the basis for the preparation of
pedigree chart using the appropriate symbols. A careful examination of the
pedigree chart would suggest whether the gene for the character is autosome-
linked dominant or recessive, X - chromosome linked dominant or recessive,
Y- chromosome linked or not.
Explanation
1. Autosome Linked Dominant traits: These are the traits whose
encoding gene is present on any one of the autosomes, and the wild-
type allele is recessive to its mutant allele, i.e., the mutant allele is
dominant.
The pedigree-chart can be of the undernoted pattern (Fig. 11.2), where
the female being interviewed is exhibiting the trait, and is indicated by
an arrow-mark in the chart.
40
EXERCISE 11
2. Autosomal Recessive trait: These are the traits whose mutant allele is
recessive to its wild type allele.
The pedigree chart can be more or less of the pattern given below (Fig.
11.3), where the lady (marked by the arrow) is showing the trait. The bar
41
L ABORATORY MANUAL: BIOLOGY
3. X-Linked Dominant traits: These are the traits whose encoding gene
is present on the X- chromosome, and the mutant allele of which is
dominant over its wild-type allele.
Such traits are very rare, and are almost difficult to find in the
population. One example is oral-facial-digital syndrome (Duchene
Muscular Dystrophy), which results in absence of teeth, cleft (bifid) tongue
associated with mental retardation. The pedigree chart may appear as
follows (Fig. 11.4):
42
EXERCISE 11
Here, the dominant mutant allele is denoted by ‘D’, and its recessive wild
type allele is denoted by ‘d’. Remember that human females have two
X-chromosomes (XX), and the males have only one X and one Y chromosome.
Males receive their lone X-chromosome from their mother, and the
Y-chromosomes from their father, whereas females receives one of her
X-chromosome from her mother, and the other X from her father.
The characteristics of such inheritance are:
(a) The trait appears in almost all the generations, and the inheritance is
vertical.
(b) If the female is affected, then about half of her sons are affected.
(c) If the male is affected then all of his daughters would be affected, but
none of his sons are affected.
(d) In short, the pedigree resembles the pattern of inheritance of autosomal
dominants, except that there is no male-to-male transmission.
4. X-linked Recessive traits: These are the traits whose encoding gene is
present on the X-chromosome and its mutant allele is recessive to its
wild-type allele.
Red-green colour blindness and hemophilia, are some of its well known
examples. The characteristic features of such inheritance are:
(a) Females express the trait only when they are homozygous for the
mutant allele, whereas the males do so even when they are hemizygous
for it.
The pedigree chart would appear as the following one (Fig. 11.6):
43
L ABORATORY MANUAL: BIOLOGY
(b) About half of the sons of the carrier (heterozygous for the trait) females
are affected. In case of homozygous females showing the trait, fifty
percent of her daughters and all of her sons are likely to be affected.
Therefore, the males are most affected in the population.
(c) Affected persons are related to one another through the maternal side
of their family.
(d) Any evidence of male-to-male transmission of the trait rules out the
X- linked inheritance.
5. Y-chromosome linked traits: These are the traits whose gene is present
on the Y-chromosome. The females do not have any Y-chromosome,
whereas all the males must have a Y-chromosome to be a male, and this
Y-chromosome they get from their father. Therefore, any trait linked to
the Y- chromosome must be present only in males, and certainly not in
any of the females. This is why these traits are also called male-sex limited
traits. All the sons of the affected male would express the trait whereas
none of his daughters would do so.
The pattern of the pedigree chart would be as follows (Fig 11.7):
Questions
1. How will you differentiate between autosome linked dominant and sex chromosome
linked dominant pedigree chart? Explain.
2. Discuss the differences in the patterns of autosome linked recessive and sex-
chromosome linked pedigree.
44
Exercise 12
Aim: To perform emasculation, bagging and tagging for controlled pollination
Principle: Conventional plant breeding programmes involve bringing under human control
reproductive processes that lead to seed and fruit formation. For this controlled pollination is
d
desirable using male and female parent having desired traits. One of the process that can be easily
brought under human control is emasculation. For this the knowledge of flower structure,
he
mechanism of pollination, fertilisation and physiology of flowering is essential for this. In
emasculation technique the stamens are removed before anthesis to obtain female parent and pollen
from the desired male parent is transferred on to its stigma.
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is
Requirement: Ornamental plants/ wild plants bearing large bisexual flower, magnifying lens,
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tweezers, small sharp scissors, brush, alcohol, rubber bands, paper bags, paper clips and tags
bl
E
Procedure
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(i) Select a flower in bud condition where antheses has not occurred. Open
the bud carefully and remove the stamens (Fig. 12.1). Mark this as
to N
d
he
Fig. 12.2 Bagging of an emasculated flower
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is
re R
(iii) Bring into physical contact anthers of a
Questions
no
46
Exercise 13
Aim: Staining of nucleic acid by acetocarmine
Principle: Acetocarmine combines with nucleic acid present in the nuclei of cells to form a deep
red conjugate.
d
Requirements: Onion bulb, onion root tips, 2 to 4% acetocarmine/acetoorcein stains, slide
he
and coverslips, brush/needle, pair of fine scissors, filter paper and microscope
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is
Procedure
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(i) Peel off epidermis from the fleshy leaf of onion and put it on a slide.
bl
Add a few drops of water over it to avoid desication.
(ii) Cut out a small piece (about 0.5 cm size) of the epidermal peel and
E
discard the remaining portion.
(iii) Wipe out the water with a filter paper.
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(iv) Put 2 drops of acetocarmine on the epidermal peel and heat gently on
a spirit lamp.
to N
(v) Apply a coverslip over the peel avoiding air bubbles and wrinkles of
the material.
©
(vi) Wipe out the excess stain with help of blotting paper.
(vii) Examine the material under low magnification of a microscope.
Observation
Record your observations with regard to shape of cell, the number of nuclei
and their position in the cell. Draw a diagram based on your preparation
and label its parts.
t
Discussion
no
Nuclei in cells are extremely rich in nucleic acid which exist in a conjugated
form with protein to form nucleoproteinous structures, called chromatin
fibres/chromosomes.
L ABORATORY MANUAL: BIOLOGY
Questions
1. What are the building blocks of the nucleic acid?
2. What is DNA and how is it different from RNA?
3. Name different nitrogenous bases present in the nucleic acid.
d
he
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is
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bl
E
be C
t to N
©
no
48
Exercise 14
Aim: To identify common disease-causing organisms and the symptoms of the diseases
Principle: There are quite a large number of organisms that are parasitic/pathogenic to humans.
These organisms substantialy damage the human body and cause diseases, which may even be fatal
d
sometimes. These organisms exhibit characteristic features in their external morphology. Symptoms
of the diseases caused by them are also specific.
he
Requirement: Preserved specimens/permanent slides/photographs of Ascaris, Entamoeba,
Plasmodium, Ring-worm fungus and compound microscope
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is
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Procedure
bl
Observe the preserved specimens/slides/photographs and note down the
E
features in the practical record book. Take care to observe all the minute
details and draw labelled diagrams of the pathogens.
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Observation
to N
A. Entamoeba
©
Systematic position
Phylum – Protozoa
Class – Rhizopoda
Type – Entamoeba histolytica
d
* Distinctive feature of the pathogen
he
B. Plasmodium vivax
(i) It is an intracellular endoparasite seen easily within the RBC of the
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infected person.
(ii) It is unicellular.
re R
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(iii) The most diagnostic stage of the parasite is "signet ring" stage in the
erythrocytes, within which it appears as a rounded
E
body (Fig. 14.2).
(iv) It has a big vacuole inside, and the cytoplasm is accumulated at one
be C
Search the stage in the blood film slide, find the signet-ring stage, and
draw its labeled diagram.
Note: It is a protozoan parasite causing malaria in humans. When an infected
©
Systematic position
t
Phylum – Protozoa
no
Class – Sporozoa
Type – Plasmodium vivax
50
EXERCISE 14
C. Ascaris
The external features of round worm are as follows: Mouth
(i) Body long (20 to 40 cm), cylindrical (5 to 6 mm
diameter) with no segmentation (Fig. 14.3).
(ii) Sexes are separate; the females are longer than
the males.
(iii) Both the ends are pointed; posterior end of male
is ventrally curved.
d
(iv) Mouth is situated at the anterior end, and is Female genital
surrounded by three lips, one present mid-
he
aperture
dorsally and rest two lips are situated
ventrolaterally (for viewing these lips a magnifying
lens is needed).
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Penial spicule
(v) Single longitudinal lines are present on the dorsal,
ventral and on the two lateral sides, all along the
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length of the body. Out of these the lateral lines
are comparatively more distinct than the others
E
lines. (b)
(vi) Excretory pore is present on the ventral surface
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Systematic position
Phylum – Aschelminthes
Class – Nematoda
Type – Ascaris lumbricoides
t
Note: Round worm or Ascaris is one of the common parasite found in the
no
51
L ABORATORY MANUAL: BIOLOGY
Systematic position
Kingdom – Fungi
d
Class – Deuteromycetes
he
Type – Trichophyton rubrum
Symptoms
Ringworm is a contagious fungal infection of the skin. Infected area of skin
pu T
is
is itchy, red, raised, scaly patches (with sharply defined edges). It is more
red on the periphery than in the center creating a ring like appearance.
re R
bl
E
be C
t to N
©
no
52
Exercise 15
Aim: To study the texture of soil samples
Principle: Texture is one of the most important physical properties of soil. The soil texture is based
upon division of the size of soil particles into three size fractions viz., Sand (2–0.05mm average
particle diameter), Silt (0.05–0.002mm) and Clay ( less then 0.002mm). If one of these fractions
d
dominates the properties of a soil, the name of that fraction is included in the name of the texture.
he
A soil which has all of these fractions in nearly equal proportion is called a loam soil.
The four terms—sand, silt, clay and
loam— are combined in various ways to name
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is
12 different textural classes. The 12 textural
classes and the percentages of sand, silt and
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clay fractions that are included in each are
shown in textural triangle (Fig. 15.1).
E
Texture affects several physicochemical
properties of soil like density, capillary and
be C
root penetration.
Procedure
Three methods are suggested here. Any one of these may be followed.
Method I
t
(i) Collect about 300–500g of soil from two different locations. Label them as sample A and B.
no
(ii) Dry the samples in an oven, or stove or in sun to remove the soil moisture (capillary and
bound water).
(iii) Select the 3 sieves of different mesh sizes (2mm, 0.05mm and 0.002mm). Arrange them in
a collecting chamber as shown in Fig. 15.2.
(iv) Place 200g of the soil in the Ist sieve (sieve of 2mm mesh) and close the lid. To sieve the soil,
shake the set manually for 5–10 minutes and collect the three soil fractions.
L ABORATORY MANUAL: BIOLOGY
d
0.05mm mesh
Observations
he
0.002mm mesh
Calculate the percentages of the various soil fractions
and tabulate:
is
fractions.
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Collecting chamber
Use the textural triangle now. Note that the three
the precentage lines of silt run parallel to the clay side of the triangle and, (iii)
perentage lines of sand run parallel to the silt silde of the triangle. In reading
to N
the textural triangle, any two particle fractions will locate the textural class at
the point where these two intersect.
©
B
t
Note for Teachers: The sieve sets contain a number and an abbreviation BSS/ASTM/
no
ISS on each sieve. In the given table (Table No. 15.1) the corresponding aperture size of
the sieves is listed. For example, BSS 30 sieve aperture size will be 500 microns.
54
EXERCISE 15
Appendix 1
1 4 5 480 4.75 mm
2 5 6 340 3.35 mm
3 6 7 280 2.80 mm
d
4 7 8 240 2.36 mm
5 8 10 200 2.00 mm
he
6 10 12 170 1.70 mm
7 12 14 140 1.40 mm
8 14 16 120 1.18 mm
pu T
is
9 16 18 100 1.00 mm
re R
10 18 20 85 850 micron
11
bl
22 25 70 710 micron
E
12 25 30 60 600 micron
13 30 35 50 500 micron
be C
14 36 40 40 425 micron
15 44 45 35 355 micron
16 52 50 30 300 micron
to N
17 60 60 25 250 micron
18 72 70 20 212 micron
©
19 85 80 18 180 micron
20 100 100 15 150 micron
21 120 120 12 125 micron
22 150 140 10 106 micron
23 170 170 9 90 micron
24 200 200 8 75 micron
25 240 230 6 63 micron
26 300 270 5 53 micron
t
55
L ABORATORY MANUAL: BIOLOGY
Method II
Texture by Feel
The texture of the soil sample can also be estimated by feeling it in the dry,
moist and wet states. Sand is coarse and gritty, silt feels smooth like flour
and clay is sticky and plastic. The smallest soil particles that one can see are
coarse silt. Feel the known texture samples first, then feel the unknown ones
and decide their textures.
d
Procedure
he
(i) Feel the dry soil first. Does it crumble easily or is it hard to break?
Hard soil samples contain a moderate amount of clay.
(ii) Take in your palm a lump of soil sample about the size of a one-rupee
pu T
is
coin and wet it to the consistency of modeling clay. Try to press it into
a ribbon between the thumb and forefinger. An alternate test is to form
re R
a wire by rolling the wet soil until it is about 1/8" in diameter.
bl
(iii) If a long wire or ribbon can be formed readily the soil is plastic and
E
probably contains over 40% of clay. Its texture must therefore be clay/
silty clay/or sandy clay. If a ribbon or wire can be formed easily but
be C
also breaks easily, the soil sample is probably a clay loam/silty clay
loam/or sandy clay loam. A heavy loam/silt loam/or sandy loam
to N
sample may form ribbon or a wire if the moisture content is just right
but these will be still weaker than the ribbons and wires formed by the
clay loam samples.
©
(iv) Next determine whether sand or silt is dominant. If there is a gritty feel
without the smooth floury touch of silt, choose a texture-name that
includes the word ‘sandy’. If the smooth floury feel predominates and
there is not much gritty feel, choose one of the ‘silty’ texture names.
Use the name without a prefix if neither smoothness nor grittiness
predominates (simply clay or sand or silt). Often this can best be
determined by adding more water until the soil is in a wet state.
If the soil is very sandy, you must choose between sandy loam, loamy
t
sand and sand. In the moist state, sandy loam samples will have some
no
tendency to stick together but loam sand and sand samples will not do so.
Use the wet state to determine whether a sample is sand or loamy sand.
After handling wet sand, your hands will be moist but clean loamy sand will
make the hands slightly soiled.
56
EXERCISE 15
Method III
Requirements: Soil samples, balance, weights, glass rod, standard sieves of
2mm and 0.5mm mesh size, blotting sheets/old newspapers, evaporating
dish and water
Procedure
(i) Collect 200-300g of soil samples from different sites, and dry them as
suggested previously to remove the moisture.
(ii) Sieve the sample through a 2mm sieve to remove stones, pebbles,
d
roots etc.
(iii) Take 100-150 g of the sieved soil sample and further sieve it through a
he
0.05 mm sieve to separate the sand fraction (collected in the sieve) from
silt and clay (collected on a blotting sheet). Weigh the amount of sand
fraction and silt + clay fraction.
pu T
is
(iv) Take a large evaporating dish (a shallow clay plate, glass trough or a
shallow iron plate) and record its weight.
re R
(v) Add the clay and silt fraction to the dish and note the weight.
bl
(vi) Add water to the dish leaving half an inch space empty at the top and
E
stir the liquid thoroughly with a glass rod taking care that the contents
do not spill out. Allow it to stand for several hours. Decant off the cloudy
be C
supernatant liquid (clay fraction). Repeat the process three to four times
until the decanted liquid is quite clear.
(vii) Dry the silt left in the evaporating dish to dryness. Cool the dish and
to N
weigh it.
Observation
©
A B
Subtract the weight of silt fraction from the weight of silt + clay fraction. The
difference will be the weight of clay decanted.
Calculate the % of sand, silt and clay fraction of the soil and express the texture.
57
L ABORATORY MANUAL: BIOLOGY
Discussion
Correlate the texture with the plants growing in the area from which the soil
sample has been collected. Discuss how the texture of soil can affect the root
penetration, tillage, soil aeration, moisture content, water holding capacity
and other aspects related to plant growth. In sandy soil the non-capillary
pore spaces will be more and the capillary pore spaces will be less. The
condition will be reverse in case of clay soil. The pore space in turn determines
water holding capacity, percolation rate, aeration, root penetration and soil
flora and fauna. Clay particles are anionic colloides and adsorb mineral
d
nutrients and minimise their leaching.
he
Questions
pu T
1. Which type of soil is better for root-penetration and better aeration?
is
2. Among sandy and clay soil which one has higher water holding capacity? Explain.
re R
3. If the clay content is high, will it affect soil fertility? Explain.
bl
4. Which type soil has poor nutrient status and high leaching?
E
5. What kind of plants grow in smooth texture soil? Name two plants that grow in
heavy-textured soil.
be C
t to N
©
no
58
Exercise 16
Aim: To determine the water-holding capacity of soils
Principle: Water holding capacity of the soil is the amount of water retained in the capillary
spaces of the soil after the percolation of gravitational water into the deeper layers. Water holding
d
capacity depends upon the capillary pore spaces in the soil. Sandy soil has very low water holding
capacity, whereas clayey soils have very high water holding capacity.
he
Requirement: Soil samples from different sites (garden, road side, bank of river, paddy field
etc.), Gooch crucible (china clay crucible with perforated bottom), filter-paper, pestle and
pu T
mortar, petridish, beaker, glass rod, balance and blotting paper
is
re R
Procedure
bl
E
(i) Dig a small pit about 10cm x 10cm x 10cm, Scoop 100–300 g of soil
from the pit and collect it in a small polythene bag.
be C
(ii) Remove the pebbles and large lumps from the soil sample.
to N
(iii) Pass the soil through a coarse sieve to remove small lumps and dead
decaying leaves and twigs.
(iv) Spread the soil into a thin layer on a sheet of blotting paper or old
newspaper and sun dry it for 2–3 hours or dry it in a pan kept on
©
stove. Alternatively dry the soil sample in oven at 1080C for 1 hour.
(v) With the help of pestle and mortar grind the sample into fine powder.
(vi) Put a small disc of blotting paper at the base of the Gooch crucible.
Weigh the crucible along with the blotting paper and note its weight.
(vii) Transfer the soil sample into the crucible. Tap the rim of the crucible
gently several times with the help of glass rod so that soil is compactly
filled and forms a uniform layer at the top. Add more soil if necessary.
t
(viii) Weigh the crucible along with soil sample and note its weight.
no
(ix) Fill the petridish with water and place two small glass rods in it parallel
to and at a small distance from each other.
(x) Place the crucible on the two glass rods in such a manner that its
bottom is in contact with water.
(xi) Leave the set up undisturbed till water appears at the upper surface of
the soil. Wait till entire soil surface is wet.
L ABORATORY MANUAL: BIOLOGY
(xii) Remove the crucible and allow all the gravitational water to flow out
from the bottom. When no more water percolates, wipe the bottom dry
with the blotting paper.
(xiii) Weigh the crucible and note its weight.
Observation
Record your observation in the following table.
Calculate the % water holding capacity of the soil as follows.
d
Weight of crucible + blotting paper: Ag
Weight of crucible + blotting paper
he
+ soil sample before experiment: Bg
Weight of dry soil: B - A= Cg
is
+ wet soil sample after experiment: Dg
re R
bl
Weight of wet soil after the experiment: D - A= Eg
Mass of water absorbed by soil: E - C= Ng
E
% Water holding capacity:
be C
Sample No. Wt. of Crucible Wt. of Crucible Wt. of soil Wt. of crucible Wt. of wet Amount of % water
+ blotting + blotting paper sample + blotting paper soil (D-A) water holding
paper (A) + soil sample (B) (B-A) = (C) + wet soil (D) = E absorbed capacity
(E-C) = N
©
A Garden
soil
B Road
side soil
C……...
t
no
D……...
60
EXERCISE 16
Discussion
Compare % water holding capacity of soil collected from different habitat
conditions. The variation in water holding capacity is due to varying
proportion of sand, silt and clay in the soil of different habitats. Soil with
very high proportion of sand have very low water holding capacity due to
large pore spaces between the particles which enables the water to percolate
freely into deeper layers leaving upper layers practically dry. In clay soil,
due to very small size of the pore spaces (fine capillaries) the water is retained
in the capillary spaces as capillary water. In these soil the water does not
d
percolate freely. Soil with more or less equal proportion of sand, silt and clay
(loam soil) combines the properties of sand and clay and therefore has
he
optimum water holding capacity and optimum soil-air for root growth.
Questions
pu T
is
re R
1. What are heavy soil and light soil?
bl
2. Give examples of a plant seen in heavy soil and light soil.
E
3. How does pore space determine the % water holding capacity of soil?
4. Why is clay soil often referred to as physiologically dry soil?
be C
7. Dead decomposed organic matter is usually added in the fields before the
cultivation of crops. Apart from providing the mineral nutrients, what additional
role does organic matter play in the cultivation of crop plants?
t ©
no
61
Exercise 17
Aim: To study the ecological adaptations in plants living in xeric and hydric conditions
Principle: Successful adjustment of plants and animals under prevailing environmental conditions
is known as adaptation. For terrestrial plants, the habitats vary from extremely dry conditions as in
d
deserts to extremely wet conditions as in marsh lands. For aquatic plants the habitats may vary from
deep water bodies like oceans and lakes to shallow ponds and pools. The plants are adapted to
he
diurnal, seasonal or annual fluctuations of the habitat conditions. For land plants the main limiting
factor is the availability of soil water whereas, for aquatic plants the main limiting factors are the
fluctuations in water level, availability of gases like CO2 and O2 and the light intensity. Adaptation
pu T
is
of land plants are primarily for conservation of available soil water, avoidance of bright sunlight
and intense heat and for aquatic plants, adaptation are for conservation of gases and efficient
re R
bl
utilization of available sunlight.
On the basis of availability of water, plants are classified as:
E
(a) Xerophytes: These are plants growing in extreme dry conditions throughout the year. For
be C
(b) Mesophytes: These are plants growing in soils with optimum soil water conditions prevailing
for major part of the year.
(c) Hydrophytes: These are aquatic plants growing in fresh to marine water.
©
The morphological, anatomical and physiological attributes of terrestrial plants are different
from the aquatic plants.
Requirement: Plant specimens from xeric and hydric habitat conditions. The specimens from
xeric condition may include a few cacti, succulents (Euphorbia, Bryophyllum, Kelancho) cycas
leaves, pine needles, twigs of Acacia, Nerium, Parkinsonia, Casuarina etc. The aquatic plants:
Salvinia, Eichornia, Pistia, Hydrilla, Vallisnaria, Utricularia, Lymnophila; some reeds like Typha,
Phragmites, amphibious plants like Marsilea and halophyte like Rhizophora. Beakers, glassjars,
t
no
Procedure
Prepare temporary stained transverse sections of leaf, stem and root of the
specimens. Study the morphological and anatomical features of the plants
EXERCISE 17
collected and look for the following adaptations. Write the name of the plant
in which a particular adaptation is observed.
Observations
Record your observation in the given tables:
d
Xerophytes
he
(Morphological/Anatomical) specimen collected)
pu T
is
b. Petiole modified into leaf like
structure
re R
c. Stem reduced, branching sparse
63
L ABORATORY MANUAL: BIOLOGY
Hydrophytes
d
d. Leaf blade pale green in colour,
finely dissected
he
e. Leaf blade waxy with thin cuticle
pu T
is
b. Stomata present on upper surface
of leaves
re R
bl
3. Absorption of water
a. Poorly developed roots
b. Root hairs absent
E
c. Roots with air pocket to help in
buoyancy
be C
b. Stem
c. Petiole
d. Leaf
5. Mechanical tissues
a. Poorly developed xylem
b. Poorly developed sclerenchyma
c. Sclerides present
Questions
t
no
64
Exercise 18
Aim: To study the adaptations in animals living in xeric and hydric conditions
Principle: The aquatic ecosystem exhibits a different pattern of abiotic factors as compared to those
in terrestrial ecosystem. The temperature of the water, penetration of sun light, the physicochemical
characteristics of water body affect the growth and survival of the biotic community. In order to over-
d
come the cumulatory effects of these factors, certain morphological and anatomical features, as well
he
as physiological processes develop in the organisms. These modifications in animals are called adaptive
features. We will study adaptations in selected animals living in aquatic and xeric condition.
pu T
Requirement: Animal specimen/models of xeric (rat, camel, squirrel) and hydric (fish, frog,
is
prawn, etc.) conditions
re R
bl
E
Procedure
Observe the animals provided and note down their adaptive features in the
be C
Observations
Hydric adaptations
©
(a) Position
Mouth (b) Presence of teeth
a. Upper jaw
b. Lower jaw
d
(a) Gills/lungs
Respiratory organs
(b) Cutaneous
he
Xeric adaptations
pu T
is
Features Adaptations
re R
Moisture getting (a) Preference for juices as food
66
EXERCISE 18
Discussion
You may have noticed many features in the body of aquatic animals
which support their life. As the different aquatic bodies vary to a
great extent, there are many other adaptive features you may notice.
For example the aquatic organism in ponds, lakes, river and sea.
Questions
d
1. Name the features that helps a frog for aquatic life.
2. What are the adaptations present in xeric animals for conservation of water?
he
pu T
is
re R
bl
E
be C
t to N
©
no
67
Exercise 19
Aim: To determine the pH of different water and soil samples
Principle: The pH value of a water/soil sample can be determined by (i) indicator dye method,
(ii) electrometric method using a pH meter and (iii) colorimetric method. For routine purposes
d
the indicator dye method using universal pH indicator solution (containing a wide range of pH
indicator dyes) or paper strips containing the pH indicators are preferred though it is not as
he
accurate as the electrometric method.
Requirement: Soil or water samples A, B and C collected from different sites (for example
pu T
soils from road side, garden, humus rich sites; water samples from borewell, handpump, pond,
is
sewage), balance, weights, filter paper, distilled water, measuring cylinder (50 mL), droppers,
re R
cavity tile, funnel, beakers (100 mL), funnel stand, universal pH indicator solution and
bl
pH indicator paper (narrow range and broad range)
E
be C
Procedure
(i) Weigh 10 g of the soil sample A. Add 50 mL of distilled water to soil
to N
(iii) Take a clean dry porcelain cavity tile. Place 5 drops of soil solution A
in three cavities of the tile as shown in Fig. 19.1.
Soil - A Soil - B Soil - C
Universal pH indicator
solution
t
pH indicator paper
no
(Broad range)
pH indicator paper
(Narrow range)
(iv) To the 5 drops of soil solution present in one cavity add 5 drops of
universal pH indicator solution. Note the colour developed and compare
it with the colour chart given on the universal pH indicator
solution bottle.
(v) To the soil solution present in the second cavity, dip a small strip of broad
range pH indicator paper (pH 2-11). Note the colour and compare with
the colour chart given on the broad range indicator paper and get a rough
estimate of pH of the sample solution.
(vi) Choose a suitable narrow range pH indicator paper (for e.g. If the pH of
d
soil is determined by you as 8.0, choose a narrow range 7.0 to 9.0) and
he
dip a small strip of it in the soil solution present in the third cavity. Note
the colour developed and determine the pH to the nearest possible value
with the help of the colour chart.
pu T
is
Repeat the same steps for determining the pH of sample B and C. Follow the
same procedure for water samples collected from different sites.
re R
Observation
bl
E
Record your observations in the given table.
be C
A B C
©
Discussion
Based on the pH values obtained, categorise the samples into acidic, basic,
t
neutral type.
no
Record the plant species present in the site from which the samples are collected.
Note for teachers: The colour developed should be noted against direct sun
light. Also, sometimes the soil solution colour may interfere with the readings.
Thus one has to be careful while making the observations.
69
L ABORATORY MANUAL: BIOLOGY
Questions
1. What will be the pH of chalk (calcareous) soil?
2. pH measurement with indicator paper is not very accurate. Comment.
3. Water logged soils are acidic. Comment.
4. Why are soil around mineral mining areas acidic?
d
he
pu T
is
re R
bl
E
be C
t to N
©
no
70
Exercise 20
Aim: To study turbidity of water samples
Principle: Various characters that control the quality of water are taste, smell, colour, amount of
dissolved nutrients, dissolved O2 and CO2, pH and different types of plants and animals and their
d
density. Turbidity of the water body determines the depth upto which light can penetrate and thus
affects the distribution and photosynthesis of phytoplanktons and macrophytes. More turbid the
he
water body less is the thickness of its photic zone.
In polluted water bodies turbidity is due to:
1. Effluents: A water body which receives domestic sewage, run off from adjacent agricultural
pu T
is
fields and liquid wastes from nearby small and large industries remains turbid.
re R
2. Planktons: A water body may be turbid due to very high density of phytoplanktons and
bl
zooplanktons, especially when the water body is rich in nutrients.
E
I. Secchi’s Disc method
be C
Requirement: Secchi’s Disc, rope of moderate thickness, meter rod, black and white
paints; paint br ush. Prepare a Secchi's disc by taking an iron disc of about 6 inches diameter, to
to N
which a weight is attached in the centre on one side and an iron hook on the other side. Tie a plastic
rope of sufficient length to the hook. Divide the upper surface of the disc into 4 equal segments and
paint two of these white and the other two segments black in such a way that black and white
©
Procedure
(i) Visit a nearby pond.
(ii) Reach to the center of the pond in a small boat.
t
(iii) Slowly immerse the Secchi disc into water vertically holding
no
the rope tightly in the hand till the black and white segments
of the disc just begin to disappear. On reaching to a
particular depth, the disc becomes completely invisible.
Mark the length of the rope when the disc just disappears
(say A cm).
Fig . 20.1 A S ecchi’s Disc
L ABORATORY MANUAL: BIOLOGY
(iv) Slowly pull up the disc and find out the length of the rope where the
black and white segments of the disc just reappear (say B cm).
(v) Find out the mean length (X) of the rope by the following method.
d
Water body Depth at which Disc Depth at which Depth of Photic
he
disappears (A cm) disc reappears (B cm) zone
Pond Site 1
pu T
is
Site 2
Site 3
re R
-
bl
E
be C
-
to N
Observations
The value X represents the depth of the photic zone upto which sunlight
penetrates in the water body and photosynthesis takes place.
©
Discussion
Greater the value of 'X' less turbid is the water. In crystal clear deep lakes, the
value of 'X' will be very high indicating, thereby, that the water body does not
have large quantities of flocculating silt or organic matter residues. This may
be due to no discharge of effluents or domestic sewage into the water body.
The high clarity of water is also an indication of very less density of phyto and
zooplanktons. These water bodies are called as non-productive or oligotrophic,
t
Precautions
Students are advised to perform this experiment under the strict supervision
of teacher to prevent incidents due to drowning.
72
EXERCISE 20
Procedure
(i) Collect about 2 liters each water samples from different sources.
(ii) Transfer 500ml of water sample in the measuring cylinders of same
volume and height.
d
(iii) Mark the three cylinders A, B and C and leave them undisturbed
overnight.
he
Observations
Observe the amount of sediment settled at the bottom of each cylinder and
pu T
is
also note whether the water above the sediment is still turbid.
re R
Record your observations in the following table:
bl
E
Water sample Thickness of sediment Clarity of water—turbid/
semiturbid/clear
be C
'A'
to N
'B'
'C'
©
Discussion
z Do all the samples show same amount of sediments?
z Which sample shows maximum sedimentation and correlate it with the
source of the sample?
z Find out whether in all the cylinders, water above the sediment is clear or
t
73
L ABORATORY MANUAL: BIOLOGY
Questions
1. Is turbid water fit for drinking? Explain.
2. Why is the penetration of sunlight in any water body important?
3. Green plants are seen only in photic zone. Comment.
4. It is a common practice to use alum for clearing turbid waters. Explain.
5. Turbidity of water body varies with season. Comment.
d
he
pu T
is
re R
bl
E
be C
t to N
©
no
74
Exercise 21
Aim: To analyse living organisms in water samples
Principle: The productivity and the trophic status of a water body is determined by assessing the
number and type of organisms (micro as well as macro) present in the water body. Water body with
d
very high density of phytoplankton per unit area is a productive water body. Such water bodies are
usually turbid and have high amounts of nutrients and dissolved oxygen. These water bodies support
he
fairly large number of organisms of different trophic levels. This is in contrast to non-productive
water bodies, which have very low density of organisms per unit area, fairly transparent waters with
low mineral concentration and dissolved oxygen and also fewer trophic levels. The status of health
pu T
is
of a water body can be determined by analysing water samples for the number and type of organisms
present in it at a given time. Such assays also help us to find out whether a water body is polluted as
re R
bl
some of the organisms are strong indicators of water pollution.
E
Requirement: Water samples from different water bodies (lake, pond, river etc.), beakers,
a few vials or small test tubes, slides and cover slips, watch glasses, dropper, compound
be C
microscope and 5% FAA (Formalin Aceto Alcohol 5:5:90:Formalin: Acetic acid: Ethanol) as
preservative
to N
Procedure
©
(i) Collect about a liter of water sample from nearby water body (pond lake,
reservoir, river etc).
(ii) Add about 5 ml of FAA to fix and preserve the living organisms present in
each sample at the place of collection.
(iii) In the laboratory, transfer the water sample into a measuring cylinder of
one litre capacity. Label each water sample to indicate the site from which
the water sample has been collected.
t
(v) Decant off the clear water, leaving concentrated sediment at the bottom.
(vi) Transfer the sediment into a vial or a small test tube. Cork and label each
vial for future use.
(vii) With the help of a dropper, transfer a few drops of sediment liquid from a
vial into a watch glass. Dilute the sediment with water if the sediment is
highly concentrated.
L ABORATORY MANUAL: BIOLOGY
(viii) With the help of a dropper transfer a drop of water from the watch glass
on the center of a slide and mount it. Blot the excess water using blotting
paper.
(ix) Prepare a few more slides of each water sample in the same way.
(x) Observe each slide, first under lower magnification and then under higher
magnification.
Observations
d
1. Record the different types of organisms present.
2. Count the number of organisms under each field of microscope.
he
3. Some of the commonly found organisms of water bodies are given in
Annexure 2.
pu T
Discussion
is
re R
Prepare a list of organisms observed in each water sample and make an
bl
assessment of type and density of different organisms in each water sample.
Polluted waters may contain very few types of organisms but in very high
E
density. The non-polluted waters will have large variety of organisms in
low density.
be C
Questions
to N
76
EXERCISE 21
Annexure 2
Cosmarium
d
he
Desmickum
pu T Stigeoelomium
is
re R
bl
E
be C
Spirogyra
Draparnaldiopsis
to N
©
Nitella
t
no
77
L ABORATORY MANUAL: BIOLOGY
d
Ceratium
he
Gymnodinum Peridinium
pu T
is
Calothrix
re R
bl
E
Rivularia
be C
Cyanomonas Cylindespermum
to N
©
Anabaena Gleotrichia
t
no
Scytonema Fischerella
78
EXERCISE 21
d
Chlamydomonas Volvox
he
pu T
is
re R
bl
E
be C
Coelastrum
to N
Closterium
Golenkinia
©
Pedrastrum
Hydrodictyon Chlorella
t
no
Scenedesmus Staurastrum
Ankistrodesmus
79
L ABORATORY MANUAL: BIOLOGY
Synechococcus
Gloeocapsa
Microcystis
d
he
pu T
is
re R
bl
Phormidium Oscillatoria
Synechocystis
E
be C
to N
©
Spirulina
Lyngbya Nostoc
t
no
Melosira
Chaetoceros
80
EXERCISE 21
Rhizoselina
d
Amphora
he
pu T
is
re R
bl
E
be C
to N
©
Biddulphia
Bacillaria
t
Synedra
no
Centronella
Nitzschia
81
L ABORATORY MANUAL: BIOLOGY
Navicula
d
he
pu T Somphonema
is
Fragilaria
Pleurosigma
re R
bl
E
be C
to N
©
Coscinodiseus
Skeletonema
Lauderia
t
no
Cocconeis
Asterionella
82
Exercise 22
Aim: To determine the amount of Suspended Particulate Matter (SPM) in air at different sites
in a city
d
largely as a by-product of man's action through direct or indirect effects of changes in energy
patterns, radiation levels, chemical and physical constitutions of environment and abundance of
he
organisms. Substances that cause pollution to the environment are called pollutants. They are the
residues of things that man makes, uses and throws away. These residues pollute soil, water and air.
The atmosphere in highly populated area is very rich in dust, smoke and SPM all due to vehicular
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exhausts and industrial emission.
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Requirement: A few freshly cut broad leaves, Vaseline, laboratory balance, weights, brush,
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paper clips and twine thread
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Procedure
This experiment is an outdoor activity and may be
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traffic. At spot 'X' expose each leaf of bundle 'A' by stretching the
attached thread and tie the two ends to two poles or branches of trees
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preferably at 10 feet height above ground. Keep leaves exposed for
about two hours.
(viii) After exposure at spot 'X', collect the leaves and carefully re-bundle
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exposed leaves and place them along with the string in the polythene
cover 'A'.
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bl Record your findings in the following table:
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Site Leaf bundle Weight of leaves (g) Weight of Total leaf area
sample suspended (cm2) of five
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Before After
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X 'A'
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Y 'B'
Z 'C'
(ix) Repeat the same process at spot 'Y' and 'Z' exposing leaves of 'B' and
'C' bundles respectively.
(x) At the end of the experiment, return back to the laboratory. Reweigh
each bundle of exposed leaves along with their respective polythene
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cover.
- Calculate the amount of suspended particles deposited in mg cm2 of
leaf at each spot.
- Compare the results of three different spots and interpret.
Since the weight of suspended particles will be in milligrams or even less
it is advised to use a very sensitive laboratory balance.
84
Exercise 23
Aim: To study plant population density by quadrat method
Principle: Density represents the numerical strength of a certain plant species in the community
per unit area. The number of individuals of the species in any unit area is its density. The unit area
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may be as small as 5 square cm to as large as 10 square metre depending on the size and nature of
the plant community under study. For herbaceous vegetation a metre square quadrat is normally
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used. Density which gives an idea of degree of competition is calculated as follows.
Total number of individual(s) of the species in all the sampling unit (S)
Density=
Total number of sampling units studied (Q)
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The value thus obtained is then expressed as number of individuals per unit area. When the
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measured unit area is divided by the number of individuals the average area occupied by each
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individual is obtained.
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Requirement: Cotton/nylon thread (five meters), 4 nails and a hammer
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Procedure
(i) In the selected site of study, make a 1 m X 1 m quadrat with the help of
nails and thread. Hammer the nails firmly and make sure that the
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Observations
Record the total number of species seen in the ten quadrats. This will give
an idea about the composition of the vegetation.
There will be difference in the species composition in the quadrats made in
shady areas, exposed areas with bright sunlight, dry or wet areas etc.
L ABORATORY MANUAL: BIOLOGY
Plant Quadrats employed in study & no. of Total No. of Total no. Density (D)
Species individuals in each quadrat individuals (S) of Quadrats
studied (Q)
I II III IV V VI VII VIII IX X
A 2 5 7 10 3 27 10 27/10 = 2.7
Z 1 2 4 8 3 2 20 10 20/10 = 2.0
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Discussion
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Plants growing together exhibit mutual relationships among themselves and
also with the environment. Such a group of plants in an area represent a
community. The number of individuals of a species varies from place to
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place, making it necessary to take many random sample areas for reliable
results. Density values are significant because they show relative importance
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of each species. With increasing density the competition stress increases
and the same is reflected in poor growth and lower reproductive capacity of
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the species. Data on population density are often very essential in measuring
the effects of reseeding, burning, spraying and successional changes.
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Questions
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86
Exercise 24
Aim: To study plant population frequency by quadrat method
Principle: Frequency is concerned with the degree of uniformity of the occurrence of individuals
of a species within a plant community. It is measured by noting the presence of a species in random
d
sample areas (quadrats) which are distributed as widely as possible throughout the area of study.
Frequency is the number of sampling units (as %) in which a particular species (A) occurs. The
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frequency of each species (sps. A or sps. B or sps. X etc) is expressed in percentage and is calculated
as follows.
% Frequency or Number of sampling units (quadrats) in which the species occurs
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=
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Frequency Index Total number of sampling units (quadrats) employed for the study
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Requirements: Cotton/nylon thread of 5 metres, 4 nails and a hammer
E
Procedure
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(i) In the selected site of study, make a 1 m X 1 m quadrat with the help of
nails and thread. Hammer the nails firmly and make sure that the
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known mark these as species A or B etc. and if the same species is seen in
other quadrats assign the same alphabet)
(iii) Similarly lay nine more quadrats randomly in the site of study and record
the names of individuals of each species.
(iv) Calculate the percentage frequency of occurrence using the formula given.
Observations
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Record the total number of species seen in the ten quadrats. This will give an
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C √ √ √ √ 4 4/10 100 = 40%
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Discussion
Variation in distribution of a species is caused by factors like soil conditions,
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quantity and dispersal of gemmules, vegetative propagation, grazing, predation,
diseases and other biotic activities. Also frequency values differ in different
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communities. They are influenced by micro-habitat conditions, topography,
soil and many other environmental characteristics. Thus unless frequency is
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not correlated with other characters such as density, frequency alone does not
give correct idea of the distribution of a species.
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species having low cover or population density also rate low in frequency, but
some may have high frequency because of their uniform distribution. Usually
if the cover and population density are high, the frequency will be high. The
plants with high frequency are wide in distribution.
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Questions
1. If frequency of a plant is high, what will be your interpretation?
2. Can many micro-habitat in an area affect frequency of a species? Comment.
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88
Exercise 25
Aim: Study of homologous and analogous organs in plants and animals
Principle: In plants and animals there are several organs or parts thereof, apparently alike in
their function and appearance, but markedly different from each other in their origin and
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anatomical structure. These organs are called analogous organs, and the seeming similarity
among them is the result of convergence, that is, adaptation to similar habitat and identical
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ecological niche.
On the other hand, there are organs or parts thereof, which apparently are quite dissimilar
to each other in appearance and perform different functions, but have the same origin and
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anatomy. The differences in their function and also in their appearances are the result of
divergence, due to adaptive radiation to different habit, habitat and ecological niche. These
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organs are called homologous organs.
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Requirement: Plant specimens showing tendrils, thorns, etc., as given in the text or any other
locally available plants, a plant with normal stem, potato and onion bulb, prickly pear, specimens
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of phylloclade, cladode, wings of bird, cockroach and bat, and cervical, thoracic and lumbar
vertebrae of a mammal/lizard
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Observations
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functionally different
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(a) (b)
Fig. 25.2 (a) Tendrils of Vitis (b) Thorns of Carissa
(iii) Tendrils of baloon vine
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Tendril (Cardiospermum) and bulbils of
Agave.
Both are modifications of floral bud,
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but they perform different functions.
Tendrils help in climbing but bulbils
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are meant for reproduction
(Fig. 25.3 a & b).
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(a) (b)
Fig. 25.3 (a) Tendrils of baloon vine (b) Bulbils of Agave
(iv) Scale leaves of onion and spines
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Tendril
(i) Stem tendrils and leaf tendrils
All tendrils are analogous with one
another, being structurally and
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their origin.
Example: Tendrils of pea and
tendrils of Vitis. Tendrils of pea are
Tendril modification of leaf and in Vitis it is
(a) (b) the modification of terminal bud
Fig. 25.5 (a) Tendrils of pea (b) Tendrils of Vitis (Fig. 25.5 a & b).
90
EXERCISE 25
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analogous to modified roots (carrot, radish) as they
perform similar function of storage of food but their
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origin is different. Rhizome of ginger, potato tuber,
(a) (b)
Colocasia are stems and beetroot, radish etc. are
Fig. 25.7 (a) Phylloclade (b) Cladode of
roots. (Fig. 25.6 a & b)
ruscus
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(iv) Phylloclade, cladode and leaves
They perform the same function i.e. they
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photosynthesise but phylloclade and cladode are
modifications of stem. Phylloclade of Opuntia,
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Parkinsonia, Asparagus and leaves of any local
plant like mango are analogous organs.
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3. Homologous Organs in Animals Fig. 25.8 Fore limb of (a) human (b) bat
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more examples.
(b) rabbit
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Questions
1. Suggest examples of homologous and analogous organs other than what are
given in the manual.
2. Why are stem and leaf tendrils considered as analogous organs?
91
Investigatory Project Work
Investigations are more open-ended than practical exercises involving a search to understand
the unknown and begins with a question or a hypothesis. You are not instructed exactly what
to do, but are given only general guidance. These give you more opportunity to plan your
work. For example, you might investigate what traits you and your classmates inherit from
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your parents and forefathers (both maternal and paternal).
he
Projects are even more open-ended than investigations. These are practical investigations
carried out by an individual or a group of students. Projects are largely your own initiative. It
also requires evaluation of your findings, redefining ideas and designing further investigations.
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This may lead to evidence that enables answering the question posed at the outset. Some of
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these projects would take about few hours to complete. Other may take few weeks. Some are
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laboratory based, others involve fieldwork. Many could be carried out at home.
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Investigatory projects are part of obligatory assignment involving purely experimental
procedures so that you report on, duplicate, or adapt something that someone else has already
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discovered. It may involve some other form of investigation also. For example, you may undertake
to investigate the richness and patterns of biodiversity (flora and fauna) in your school campus
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and prepare a mural of it or to investigate the effects of physical fitness on your pulse rate.
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electronics, etc.
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short for follow-up and any fascinating secondary aspects that you may come
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across.
Designing Projects
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Having established the objectives of your chosen project, you must have an
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experimental design. This will allow you to collect the data you need in a
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scientific way to test the hypothesis. For example, if your project involves
bl
investigating the hypothesis that stale milk contains more bacteria than fresh
milk, devise the procedure you would adopt to carry out your investigation.
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Planning Investigations
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Having decided your topic for scientific investigation, you should give careful
thought to the plan of your investigation in some detail. These may include
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93
L ABORATORY MANUAL: BIOLOGY
¨ How do you plan to analyse your results? Would you employ statistical or
other methods? Are scale range, interval, number of values chosen are
adequate and reasonable ?
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type of readings or measurements you are going to make, how often and how
many. Note the source of error, if any, that you come across.
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¨ Handle instruments and equipments appropriately to give accuracy.
¨ Repeat measurement.
¨ Keep proper controls and the variables constant.
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Reporting/Writing of Project
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A format, such as given below, can be followed.
(i) Title of the investigatory project: Write the title of the project, for
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example, ‘Inheritance pattern of eye colour’.
(ii) Objectives: Express as clearly as possible the effect of one variable that
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(v) Result: A suitable chart or table for recording and organising your
readings or measurements should be made out before you start the
experiment.
(vi) Analysis and interpretation: Observation data are factual, and may
not be as expected by you.
(vii) Discussion: Discuss briefly the implication of your results and suggest
extensions of any kind that can be undertaken.
(viii) Conclusion: In view of the results obtained and related work done on
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(ix) References: Any work related to the project which you have come across
through books/articles or any other source should be written as reference,
for example: Michael Michalco (2001), Cracking Creativity, Berkeley, Ten
Speed Press.
This write up is meant to train the students in scientific methods. In other
words, it accentuates the spirit of enquiry and investigation in young minds.
94
INVESTIGATORY PROJECT WORK
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certainly give a sample of the thrill of discovery.
Following are pages on procedural guideline about a few suggestive
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investigatory project work.
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Background information
Monitoring the physico-chemical properties of water is of vital importance.
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Normal maximum permissible limit of pH for our life and health is 6.5–8.5.
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Abnormal levels of pH and their consequences are given below:
pH 3 to 5 is too acidic for most organisms to survive, when the pH of water falls
below 4.5 most of the fishes die, leaving only a small number of acid-tolerant
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insects such as water boatman and whirligig. These insects (beetles) can survive
and multiply even at pH 3.5. Similarly, pH>8.5 is too basic for most organisms
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to survive.
Materials needed
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95
L ABORATORY MANUAL: BIOLOGY
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tendency to disappear Larvae of midge (Chironomus) R A
as degree of pollution
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Blood Worm X X
increases
Leech (Hirudinea) X X
Water louse, water skaters X X
(Asellus)
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Fresh water shrimp (Gammarus) X X
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Water boatman X X
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Diving beetle (Dytiscus) X X
Caddisfly larva (Ochrotricha)
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X X
Damselfly larva (Lestes) X X
Stonyfly nymph (Isoperla)
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X X
Mayfly nymph (Stenonema) X X
Snail (Lymnaea) none X
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Utricularia X X
Chara X X
Water fern (Salvinia) X X
Water velvet (Azolla) X X
Water meal (Wolffia) X X
Lesser duckweed (Lemna minor) X X
Greater duckweed (Spirodela) X X
Diatom X X
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INVESTIGATORY PROJECT WORK
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(i) Avenue trees
(ii) Wind Breakers
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(iii) Road dividers
(iv) Sound barriers
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(v) Medicinal and other uses
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5. Agrochemicals and their effects
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The project may be carried out in a survey mode with a questionnaire prepared
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with the help of the teacher to cover the following aspects.
(i) List of pesticides used, amount used/hectare or acre, periodicity of spray,
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name of the crop plant grown, recommended dose and the dose employed,
known effects on pest, whether the chemical pesticide is biodegradable
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Note the various types of insects, birds, reptiles, amphibians, mammals, etc.,
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L ABORATORY MANUAL: BIOLOGY
(v) Predator
(vi) Vectors for transmission of diseases
(vii) Any other
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product.
(iii) Possible types of pollutants released by the industry (air/water/
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soil).
(iv) Measures undertaken by the management to comply with the standard
set by Central Pollution Control Board (CPCB), PCBs, etc.
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(v) Awareness about ISO 2000.
(vi) Impact assessment carried out or not.
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08. Study of the effect of chemicals and pollutants on the Mitotic
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Index of the mitotically dividing onion root tip cells
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INVESTIGATORY PROJECT WORK
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need and problems.
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Investigatory Project 1
Aim: To study the effect of pH on seed germination
Principle: pH is one of the most important factors that controls the composition of flora and
fauna in different terrestrial and aquatic ecosystems. pH of soils is essentially controlled by the
amount and type of various minerals and also by the quality and quantity of humus (dead,
d
decaying organic matter) present in it.
he
Seed germination is controlled by pH of the germinating medium. Seeds of different species
prefer a specific range of pH for maximum germination. pH not only controls the germination
of seeds but also growth and development, reproduction and various other metabolic activities,
of the plant.
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Objectives: After completing the project, the students will be able to
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1. Plan out an experiment and understand the use of appropriate chemicals, apparatus and
E
equipment, and learn the preparation of solutions.
2. Understand research methodology.
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Materials required:
1. 125 seeds each of sunflower, mustard, green moong, alfa alfa, fenugreek and barley (selection
of seeds of different species may be made as per their availability)
©
2. Phosphate buffers
3. Distilled water
4. Petridishes of 7.5 cm diameter (15 pairs)
5. Blotting papers cut into circular discs to the size of petridishes
Procedure
t
(ii) Wash the seeds with water and blot them dry.
(iii) Select an appropriate place in the laboratory where there is sufficient
light. Arrange petridishes in three horizontal lines, with 5 dishes in each
line. Arrange petridishes horizontally in three rows A, B and C with seven
dishes in each row.
INVESTIGATORY PROJECT 1
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(ix) Cover the petridishes and record your observation after every 24 hours
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for 7 days. Tabulate your results as given.
Observation
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Observe the emergence of radicle as an indicator of germination and record in
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the table. Calculate the percentage of germination every day.
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Table: Percentage of ................ seed germination
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No and % of seed germination Total % of seed
germination
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pH 1 2 3 4 5 6 7 A B C
4
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5
6
7
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• Find out at what pH range seeds of different species had maximum % of germination.
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Investigatory Project 2
Aim: Quantitative analysis of phytoplankton in a water body
Principle: The species composition and the density of the phytoplanktons determine the
productivity status of a water body. Phytoplanktons are the principal producers in a water
d
body. Based on the density of phytoplanktons, the water bodies are classified into non-
productive or oligotrophic and highly productive or eutrophic. Oligotrophic water bodies
he
support only a few species, whereas eutrophic water bodies support large number of species.
Further, the species composition of the phytoplanktons indicates the status of health of the
water body. Through phytoplankton assays, limnologists make an estimate of degree of pollution
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in the water body. High density of cyanophycean algae, diatoms, volvocales, etc., are the
indicators of high degree of pollution. It should be noted that density and the species composition
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of phytoplanktons exhibit diurnal, seasonal and annual fluctuations. It therefore becomes
important to monitor water bodies at regular intervals for drawing specific conclusions related
E
to their ecology.
It is in this context, the procedure for phytoplankton analysis on qualitative (species
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composition) and quantitative estimation (density/unit area) is suggested for students who
want to enter into fascinating realms of aquatic ecology.
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Procedure
(i) Plankton net resembles the butterfly net in several aspects. Plankton
net, however, is prepared from bolting silk cloth which is readily available
at shops dealing in scientific equipments and chemicals. Procure about
one metre of bolting silk cloth of 40 mesh size and stich out of it a 40 cm
INVESTIGATORY PROJECT 2
d
firmly a few centimetres above the water surface.
(iv) Immerse and fill the plastic bucket with water completely upto 15–litre –
he
graduated mark and filter the water through the plankton net. Repeat
the process several times (say10 times).
(v) At the end calculate the quantity of water in litres (X) filtered by
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multiplying the amount of water in one bucket and number of buckets
of water filtered.
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(vii) During this process of filtering, the planktons are collected in plankton
bucket. Only the water free of planktons escapes through the mesh of
E
the net.
Splash a few buckets of water against the net from outside taking care that
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no water enters into the cone from the mouth. This will wash all the planktons
sticking against the inside wall of the net into the plankton bucket.
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Detach the plankton bucket from the net and add a few drops (1–2 ml) of
5% F.A.A. to the plankton concentrate. Transfer the concentrate collected into
a suitable specimen tube and cork it. Note the volume of the concentrate (Y).
©
In the laboratory
1. With the help of 1 ml pipette, draw 1 ml of concentrate and transfer it
dropwise into the watch glass. Count the total number of drops that make
1 ml of concentrate (A).
2. Transfer one drop of plankton concentrate from the watch glass on a clean
slide. Cover it with square shaped cover slip. (For convenience divide the
area of the cover slip into parts with the help of lines drawn by Indian ink).
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Observation
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Observe the slide under microscope and count the number of total organisms
(B) by moving the slide from one corner of the cover slip to another horizontally
as well as vertically till the entire sample under the cover slip is completed.
With the help of following calculations find out the total number of different
organisms per litre of water.
103
L ABORATORY MANUAL: BIOLOGY
1000×(A BY)
Unit cells/L = Unit/L =
x
Where A = number of drops in 1ml concentrate
B = number of organisms counted in 1 drop of concentrate
X = total amount of water filtered
Y = total volume of concentrate after filtration
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density of organisms under the guidance of teacher.
he
Find out the density and composition of organisms in different water samples
(polluted/non-polluted).
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Note the common organisms in both the water samples and those specific
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to each sample.
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104
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Notes
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Notes
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