Fungi Bacteria and Virus - H C Dubey
Fungi Bacteria and Virus - H C Dubey
Fungi Bacteria and Virus - H C Dubey
H.C. DUBE
Department oj Biosciences
Sardar Patel University
Vatlabh Vidyanagar
(Gujarat)
v
VANI EDUCATIONAL BOOKS
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1985 Edition
H.C. Dube
CONTENTS
REFERENCES 225—229
INDEX 231—240
1. INTRODUCTION TO THE LIVING WORLD
In his boot “The Nature of Life”, Szent Gyorgi writes: “life as such
does iiat exist; no body has ever seen it; the noun life has no sense,
there being no such thing.” But a definition is necessary containing
everything it has to contain and excluding everything it has to ex¬
clude.
‘‘Life may be considered either as a property or as a manifestation
or as. a state of organisms.” But what are organisms? 'What is the
specific property of living organisms which is not found in nonliving
(inanimate) world?
An organism is a complex, organized, specific system able to repro¬
duce its kind.
Characteristics of Life
1. An organism always derives from a pre-existing organism. (It
never developes de novo). So organisms have a genetic continuity.
2. The organism synthesizes constituent parts from food and ener¬
gy. The organism consumes the food (i.e. metabolizes) for synthesis
and growth.
3. The organism is the independent unit of reproduction.
4. An organism is composed of macromolecules—(proteins, nucleic
acids, carbohydrates) which work in co-operation and follow the rules
of co-ordination strictly. When some molecules get rid of the control,
"“molecular diseases"” appear. Tumors (cancer) are the products of un¬
controlled growth and cell divisions.
5. Living organisms are metastable ‘maxwell demons’ whose stable
state is death.
Plantae
Animalia,
Protista
(unicellular eukaryotes!
-Monera
( = Proka,yota)g
Fig. 1.1. Five kingdoms of the living world (after Whittaker, 1969).
1. Monera—The kingdom of bacteria and blue-green algae. It in-
> *y tides all prokaryotic organisms. t >> i
JH | 2. Protista—includes the unicellular eukaryotes. ( i \>J 5 jJ*
j. Planatae or Plant kingdom—includes all green plants.
C ** 4. The Fungal kingdom.
^ 5* Animalia or the Animal kingdom.
l
Introduction to the Living World 3
Since ancient times man has loved to get the ‘kick’ by a drink,
without bothering that his cup contained the product of fermentation
caused by a fungus. It was prov¬
ed by Louis Pasteur, the great
French biologist, to whom also
goes the credit of giving the
‘germ theory of disease’, that a
chemical—(enzyme) produced by
veast cells converted the sugar
into alcohol. Since then began
the study of chemical activities
of living organisms, which is now
line of the most fascinating
branch of science; called bio-
chemistry.
The penicillin was discovered
in 1928 by Alexander Fleming of
Britain from the fungus Petti-
cillium noiatum, which in 1940’s
emerged as a ‘wonder drug’ for
the treatment of bacterial diseases. It gave another important ‘niche’
to fungi in the realm of biological sciences as producers of antibiotics.
Then, in Japan some intelligent farmers noticed that their rice
plants grew unusually tall but gave no or poor yield. The disease
was named ‘bakane’ or foolish rice disease. The tallness, associated
with paleness, was caused by a new type of growth substance called
the Gibbered ins, produced by the fungus, Gibberella fujikuri
(—Fusarium mpniliforme), which caused the ‘bakane’ disease.
Earlier, Indole Acetic Acid (IAA) was isolated from RJiizopus niger
before it could be detected in other plants.
The ‘biochemical genetics’, which later developed into the charm¬
ing ‘molecular biology’ was founded by studies with Neurospora
crassa, a fungus. It was especially suited for genetical analysis.
Neurospora dethroned Drosophila from the kingdom of genetics but
soon yielded to bacteria, which in turn, yielded to viruses. Neverthe¬
less, in 1952 Aspergillus nidulans emerged again on the scene when
Pontecorvo and Roper of Glasgow discovered a novel type of sexual
reproduction called ‘parasexual phenomenon.’ Fungi, thus contihu^
to be important organisms in genetics.
“Without fungi even death will be incomplete,” said Pasteur. The
dead cellulosic vegetation is decomposed into carbon and minerals
6 A Textbook of Fungi, Bacteria and Viruses
which are returned to the same environment from which they were
taken. Thus, fungi maintain the carbon and mineral cycles in nature.
But we have to keep them at arms length. They are great nuisance;
grow on everything from jam to leather and poison or destroy them.
The ‘hallucinogenic fungi5 have caused greatest damage by making
us perceive the sound and sight which does not exist. LSD (d-
lysergic acid diethylamide) produced from Ergot (Claviceps purpurea)
gives an unreal, extraordinary lightness and hovering sensation. It
has greatly corrupted our frustrated youth. Fungi produce interesting
metabolites which are put to innumberable uses.
Classification
The classification followed here is based on that proposed by
Ainsworth (1966), which has been followed in The Fungi—An
Advanced Treatise,(1973) edited by Ainsworth, Sparrow7 and Sussman.
Fungi; Characteristics and Classification 7
Division Division
Myxomycota Eumycota
(4 classes) (5 sub-divisions)
1. Acrasiomycetes 1. Mastigomycotina
2. Hydromyxomycetes Class—Chytridiomycetes
3. Myxomycetes Class—Hyphochytridiomycetes
A. Plasmodiophoromycetes** Class—Oo mycetes
Class—Plasmodiophoromycetes
2. Zygomycotina
Class—Zygomycetes
Class—T ri chomy cetes
3. Ascomycotina
Class—Hem i a scomypetes
Class—Loculoascomycetes
Class—Plectomycetes
Class—Laboulbeniomycetes
Class—Pyrenomycetes
Class—Discomycetes
4. Basidiomycotina
Class—Teliomycetes
Class—Hymenomycetes
Class—Gasteromycetes
5. Deuteromycotina
Class—Rlastomycetes
Class—Hyphomycetes
Class—Coelomycetes
de Bary (1831—1885)
Heinrich Anton de Bary, a German, was a medical man. In 1855 he took up*
botany and made important contributions to mycology, de Bary’s mycological
interests were more biological and physiological than taxonomic. He discovered
many things. The establishment of Phytophtkora as cause of the late blight of
potato (which ravaged the potato crop in Europe in 1840’s) and the discovery of
heteroecism stand out as the most important, de Bary is called as “The founder of
modern Mycology” and “The father of Plant Pathology”'.
Fungi: Characteristics and Classification 9“
de Bary
^General Characters
Habitat. Fungi are everywhere—in water, air and soil. They
are found in hot deserts and on high mountains; they grow on rocks,
living or dead plant and animal bodies, jams and jellies, leather,
paper and plastics. The wide distribution of fungi is because of
their potential to utilize any kind of substrate under any kind of en¬
vironment. Thus, there are the cellulolytic fungi, cutmolytic fungi
and lignin decomposing fungi, named on the basis of the substrate
decomposed. Coprophilous fungi thrive on dung of herbivores,
while keratiuaceous fungi live on the dead remains of animals.
Thallus. The thallus may be unicellular on filamentous. Fungi are
characterized by their hyphae (sing, hypha)— the long tubular, much
branched structures which collectively are called mycelium (pl=
mycelia). The hyphae may be unseptate or septate (Fig. 3.1). The
*Septa are of two types: primary and adventitous. Primary septa are formed
during mitosis or meiosis to separate the daughter nuclei. Vegetative septa, on
the other hand, are not associated with nuclear divisions but are formed to
delimit sex organs or to cut off older hyphae.
division Eumycota 11
Pseudoparenchyma Prosenchyma
finally into soluble units which are then absorbed by the hyphae.
The fungi live as parasites, saprophytes or symbionts. As parasites,
fungi grow on plants, animals and human beings causing diseases by
their presence. Fungal parasitism varies from facultative parasitism to
obligate parasitism. The facultative parasites can grow saprophy-
tically in the absence of the host while obligate parasites (downy
mildews, powdery mildews and rusts) grow only on their specific
14 A Textbook of Fungi, Bacteria and Viruses
{see Chapter 10), they are the partners with algae. In Mycorrhiza
(meaning ‘fungus roof) the fungal hyphae form a compact pseudo-
parenchymatous mantle around the roots (Fig. 3.7) of some vascular
plants like, Pinus and also extend into the cortex. The mycorrhiza
help in absorption of minerals. The fungus mantle acts as a water
sponge. Some plants won’t grow until their roots have come in touch
with the mycorrhizal fungi.
Growth. Growth involves cell enlargement, accompanied by cell
division and cytoplasm synthesis. The tip of the hyphae (50 to 100 n
in the apical region) is the zone of
elongation and is filled with proto¬
plasm (Fig. 3.8). The rest of the hypha
is vacualated and is incapable of elonga¬
tion. But it helps in growth by syn¬
thesizing the cytoplasm, which is
continuously transported to the tip.
The fungus grows only at the tip. The
growing hypha forms branches in
acropetal order behind the growing
apical region. A soft portion appears
on the wall which bulges out as a bud
and extends to form a lateral hypha.
Secondary and tertiary branches are
formed in the same way.
r* 4-u • i i . Fig. 3.8. Showing growing
Growth is measured by increase in tip of fungus hypha.
the radius of a fungus colony growing
in a Petridish or more accurately by taking the dry weight of the
ffingus grown in a liquid medium after 5, 10 or 15 days.
Reproduction. Fungi multiply by non-sexual (vegetative, and
asexual reproduction) or sexual methods of reproduction. The non-
sexual methods (fragmentation, budding, fission, chlamydospores,
oidia, zoospores, aplanospores, conidia etc.) serve to multiply the
fungus and disseminate it to far off places. The fungus extends its
family. But the function of sexual reproduction is different. It occurs
usually during conditions which are not suitable for the fungus
growth. The fungus winds up by restoring to sexual reproduction',
and undergoes hibernation or resting period.
Oidia (—Arthrospores)
and female. When the gametangia are of different shapes, these are
called heterogametangia—the male is usually smaller and club-
shaped, while the female is bigger and globular. Fusion between
and their contents come to lie in the common cell formed by their
fusion. In the other type (e.g.. Rhizophidiuni)y the contents of the
male gametangium migrate into the female gametangium through a
pore or a fertilization tube. The male gametangium is left empty.
O'O Gametangial contact. The antheridium (and occasionally also
the oogonia) is not differentiated into definite protoplasts or gametes
but are represented only by their nuclei. The male nuclei (never the
cytoplasm), migrate into the oogonium through a pore dissolved at
the point of contact or through a fertilization tube formed. by the
antheridium, example: Pythium,. Phytophthora.
(c) Spermatization. This occurs in Asco-and Basidiomycotina.
Spermatia (sing, spermatium), which are minute, male gametes, are
formed like coftidia on spermhtiophores. The spermatiophores may be
formed exogenously or arranged inside a spermogonium e.g., Puccinia.
The spermatium,when comes in contact with the female gametangium
through wind, insects, water sugary exudates etc.;, releases the male
nucleus into the female gametangium through a pore.
(d) Somatogamy. In some higher Ascomycotina and Basidio¬
mycotina, sex organs are not formed. The somatic cells,, as such, act
as gametangia. Their fusion is governed by rules of homothallism
and heterothallism.Thus somatogamy may occur betweencells of the
same hypha—(in a homothallic genus) and between cells of different
thalli (in a heterothallic fungus).. Anastomosis, which is frequent in
higher fungi, brings the male and female nuclei under one cell. Cells
of the monokaryotic hyphae of Asco-and Basidiomycotina fuse result¬
ing in plasmogamy and establishment of a binucleate cell. From this
cell the dikaryotic mycelium is formed.
PARASEXUAL PHENOMENON
Heterothallism
If the male and female gametes or the gametangia are borne on the
division Eumycota 21
LIFE CYCLE
The fungi are haploid organisms having haploid nuclei in their cells.
The thallus reproduces itself by asexual methods (Fig. 3.13). Even-
class CHYTRIDIOMYCETES
order CHYTRIDIALES
The order Chytridiales has several families. We shall study the genus
Olpidium included in the family Olpidiaceae.
family OLPIDIACEAE
In this family the thallus is endobiotic (i.e. remains inside host cell)
and holocarpic (i.e., the whole thallus is converted into a zoospo¬
rangium or a resting sporangium). Sexual reproduction is by piano-
gametic copulation which results in the formation of a biflagellate
zygote. The zygote enters the host cell and forms a resting sporan¬
gium,which on germination forms zoospores. Olpidium is a typical
example of this family.
genus OLPIDIUM
Root cell
Muitinucleate
thallus
Encystment
<en root hai
Zoosporangium
Exit tube
Isogamy
Meiosis
Zygote Karyoqamy
Resting
sporangium
Entry of protoplast
class PLASMODIOPHOROMYCETES
“The class has only one order, the Plasmodiophorales with a single
26 A Textbook of Fungi, Bacteria and Viruses'
PLASMODIOPHORA BRASSICAE
(Club-root disease of cabbage)
Germination
y/^Ftagella shed
Resting spores
in host-cell h
Infection of root hair
y
,n
root
\^ y Primary plasmodium
\ Multinucleate pr,
▼ / plasmodium
Merosis
Cleavage of
Karyogamy plasmodium
Zoosporangia
Multinucleate
secondary
plasmodium ^
^ Zoosporangia
' with zoospores
Secondary plasmodium
terior into the soil through an exit tube formed on the sporangium on
through pores formed on the root hair. The zoospores fuse (isogamy^
and form a binucleate quadriflagellate zygote. Karyogamy does not
follow plasmogamy.
Secondary phase. The zygote penetrates a root hair and enters the*
cortex, where it forms a multinucleate secondary plasmodium. Kary¬
ogamy takes place and the secondary plasmodium becomes a diploid
plasmodium. Meiosis follows and the resulting haploid nuclei multiply,
by mitotic divisions. By cleavage of the cytoplasm and wall formation*
28 A Textbook of Fungi, Bacteria and Viruses
class OOMYCETES
order SAPROLEGNIALES
family SAPROLEGNIACEAE
genus SAPROLEGNIA
Reproduction
Asexual reproduction. Asexual reproduction is accomplished by
the zoospores (Fig. 4.4). Cylindrical zoosporangia develop at the
tips of hyphae which produce numerous biflagellate zoospores. When
mature, they can be seen jostling one another within the zoospo¬
rangium. The apex of the zoosporangium suddenly breaks and the
zoospores escape. At first the zoospores come out rushing;as if dis¬
charged by pressure; but afterwards the remaining zoospores escape
slowly. A cine film of the emptying sporangium has shown that the
zoospores escape with the blunt end foremost, but as soon as the
spores are outside, their direction of motion reverses, and they swim
with the pointed end forward. The zoospores are propelled by their,
flagella, one pointing forwards and the other trailing behind.
A Textbook of Fungi, Bacteria and Viruses
Encysted
Primary zoospore
f ’W V
Prjmai^y zoospore Germination ^
/ Secondary zoospore
Zoosporangium
Empty sporangial cases
Encysted Secondary
\
zoospore
Developing sex
organs
Oogonium
Egg
Gametangial
contact
X = Pla*mogamy)
genus ACHLYA
Reproduction
Asexual reproduction. Zoospores are formed in cylindrical
zoosporangia borne on undifferentiated zoosporangiophores (Fig. 4.6). j
The primary zoospores, which are all ejected together,and not one by
one as in Saprolegnia> are turned into cysts as soon as they come out.
The cysts are arranged as a hollow ball at the tip of the sporangium.
Each cyst, after a short rest period, gives rise to a laterally-biflagel-
sub-division Mastigomycotina 33
Jate secondary zoospore. So, there is only one swarming spore in the
life cycle of Achlya.
Saprolegnia Achlya
order PERONOSPORALES
family PYTHIACEAE
genus PYTHIUM
Reproduction
Asexual reproduction. Globose or oval sporangia are borne
terminally on the hyphae (Fig. 4.8). Distinct sporangiophores are
not formed. At the time of zoospore formation, a bubble-like
vesicle with a long stalk emerges on the sporangium. The sporangial
protoplast moves into the vesicle. The zoospon s are formed in this
vesicle. This process is completed within 15 to 20 minutes. When
the crowded zoospores start rocking motion, and bounce on the wall,
the delicate vesicle bursts like a soap bubble releasing zoospores.
The zoospores, which are reniform and biflagellate, after swimming
for sometime in the film of soil-water, come to rest; encyst (J.e. with
draw the flagella and form a wall) and germinate by forming a germ
tube.
The zoosporangia in dry weather germinate directly by a germ
tube rather than by forming zoospores and thus behave like conidia.
Sexual reproduction. Sexual reproduction is oogamous and
occurs by gametangial contact. The oogonia and antheridia may be
terminal or intercalary and borne on the same or different branches.
The oogonium is globose and has a single multinucleate oosphere in
sub-division Mastigomycotina
Vesiple Vesicle bursts releasing zoospores
Zoospores
Tube
Zoospore
swarms
Encysted zoospore/jgv 0^
^Sporangiophore Germination
Somatic hypha
.Oogoniu*
Periplasm
Oospore
• Antheri;
Oosphere (egg)-fc%?J^
Haplophase (n)
hase
Fertilization tube/ /■/
!X
Karyogamy
r
fj
Gametangial contact
(Plasmogamy)
Control Measures
Damping off of seedlings and root rots caused by Pythium are more
common in soils of high water and humus content. Thus, water
logging in fields should be avoided. Soil sterilization to kill the
fungus is done through fungicides (perenox, formaldehyde or Borde¬
aux mixture). But the better method of control is to treat the seeds
before sowing with protective fungicides like organomercury com¬
pounds or copper fungicides.
genus PHYTOPHTHORA
Introduction
Phytophthora, which shows much resemblance to Pythium, however,
differs in several ways. The important differences lie in the following
characters:
1. The method of sporangia! germination (no vesicle is formed and
the zoospores are fashioned in the sporangium itself).
2. The production of distinct sporangiophores which are easily dis¬
tinguishable from the somatic hypha.
3. Sporangia are lemon-shaped with a distinct papilla.
4. The resumed growth of the sporangiophore tip pushes the
sub-division Mastigomycotina &
Reproduction
Asexual reproduction. Chlamy do spores—Terminal or intercalary
chlamydospores are abundantly formed on the hyphae.
Sporangia. A low temperature (18—20°C), accompanied by,
high relative humidity (91—100%), are the requisites for sporangial
production. Sporangiophores emerge through the stomata from
the lower surface of infected leaves (Fig. 4.9) and bear terminally,
lemon shaped sporangia with a distinct beak (Fig. 4.10). The renewed
growth of the sporangiophore gives it a sympodial zigzag shape. On
potato tubers, the sporangiophores emerge in large numbers from th-
cut surfaces. The sporangia are deciduous (fall off) and get widely
disseminated through soil water (especially irrigation water) or
through wind. Wind-disseminated sporangia, germinate directly
by germ tube and, thus, functionally become conidia. They are
highly susceptible to desiccation. In presence of water, the sporan-
Fig. 4.9. Sporangiophores and sporangia
of Phytophthora infestans hanging from
the lower surface of blighted potato leaf.
Zoospores
-Papilla
mm Zoospore^®"
Zoosporangia
Encysted
zoospore
Germination
Sporangiophore
Somatic hypha
Zoospores
Oogonium
IP^Germ Haplophase(n)
sporangium /\
Periplasm»-I__^>y
OosphereV^SS!V0o90nium
Germination
19 Oospore
/Q \ '•,^^-Antheridium
y
Pathogen—Phytophthora infestans
The disease is world wide in occurrence and is reported also from
all parts of India. Epiphytotics never occur on the plains, but are
frequent on Hills. The non-availability of high relative humidity
during the crop season (October-December), prevents the disease
occurrence. The fungus, present in potato tubers, fails to survive
the high summer temperature on the plains. The disease recurrence
occurs through ‘seeds’ (tubers) stored in cold-storages. The name
*late blight,’ is given to this disease because, compared to another
disease—‘Early blight’ (caused by Alternaria solani),it occurs late in
the season. The brown spots appear on leaves only at the time of
flowering; grow very fast and soon cover the entire foliage (Fig. 4.11).
Ultimately, the infection reaches the underground portions of the plant
and infects the tubers. Independent infection of tubers also takes
place by zoospores present in the soil.
Control Measures
1. Certified, disease-free tubers should be used for sowing.
2. Fungicides (Bordeaux mixture, Dithane D-14, Dithane Z-78
and Dithane M-22 etc.) should be sprayed regularly, right from the
seedling stage.
3. When the fungicides fail, the extreme step is taken to save the
tubers by spraying sulphuric acid on the foliage, several days before
harvesting. The foliage is killed, and along with them the fungi also
meet their end.
COLOCACIA BLIGHT
Pathogen—P. colocasiae
The disease occurs during rainy season (August-September). Cir¬
cular to irregular brown patches appear on the leaves which show
distinct zonations (Fig. 4.12). Drops of a yellow liquid appear on
the brown areas. The affected tissues dry and fall off, leaving holes
.and cuts on the surface of the leaves. The corm also gets infected
and starts rotting.
The disease perpetuates from season to season through infected
cOrms which are used as seeds.
Control
1. By spray of fungicides (Bordeaux mixture, Blitox-50 and
44 A Textbook of Fungi, Bacteria and Viruses
The family contains only one genus Albugo (=Cystopus), which is*
an obligate parasite. It is characterized by stout, club-shaped, un¬
branched sporangiophores which bear a chain of sporangia at the:
apex. Haustoria are knob-like. The oospore is plerotic i.e. comple¬
tely fills the oogonium.
genus ALBUGO (=CYSTOPUS)
knob-like haustoria.
Reproduction
Asexual reproduction. Sporangia or Conidia. After attaining
a certain stage of maturity, hyphae aggregate at several places under
the epidermis (Fig. 4.15). Club-shaped sporangiophores of deter¬
minate growth, with thick lateral walls and a thin apex, arise to
form a palisade layer. These cut multinucleate sporangia which
remain attached to form a chain at the apex. The oldest sporangia
lie at the top and youngest at the base of the chain. The walls
between the sporangia fuse to form a gelatinous disc-like structure
called disjunctor or separation disc. The disjunctors are dissolved
by water, and the sporangia are set free. The numerous sporangia
that are produced at the apical end of sporangiophores, push against
the epidermis which bulges out and ultimately breaks. When the
epidermis bursts the white mass of sporangia become visible as a
white crust on the leaf surface. These areas with broken epidermis
. and creamy mass of sporangia appear as pustules or blisters on the
leaves.
The young sporangia are globose but due to mutual pressure
become hexagonal. They are disseminated by wind. Germination
occurs when the sporangia reach a suitable host. The mode of
germination depends on the availability of water. If water is avail¬
able zoospores are formed, otherwise the sporangium germinates
directly by forming a germ tube and thus behaves like a conidium.
Mature 4—12 biflagellate and reniform zoospores are formed in the
sporangium which move into a vesicle formed on the sporangium,
before they are released. After a swimming period, the zoospores
come to rest, encyst and germinate by a germ tube. The germ tube
enters the host and establishes an inter-cellular mycelium. The
repeated production of sporangia spreads infection to more and
more plants.
Sexual reproduction. When the crop season approaches its end,
sexual reproduction ensues. The hyphae penetrate deeper into the
tissues and form sex organs. The onset of sexual reproduction is
usually indicated by hypertrophy (Fig. 4.14) of the infected part.
The sexual reproduction is by gametangial contact of dissimilar
sex organs (oogamous). There are cytological differences in the
fertilization of the various species. The following description is for
A. Candida.
The antherida and oogonia, borne terminally on somatic hyphae
46 A Textbook of Fungi, Bacteria and Viruses
Multinucleate
g3metangia
Uninucleate*
ametangia
Gametangial
contact
(Plaemogemy)
family PERONOSPORACEAE
order MUCORALES
(Pin molds or sugar fungi)
family MUCORACEAE
genus MUCOR
Economic Importance
The harmful effects of Mucor in spoilage of food, textile, leather is
well known. Less known are their beneficial activities. They are
important in the bio-degradation of complex organic materials.
Species of Mucor are used in fermentation process in industry for the
conversion of starch into glucose which is then acted upon by yeast.
Yeast does not produce the enzyme amylase and therefore it cannot
degrade starch.
50 A Textbook of FungU Bacteria and Viruses
Reproduction
Asexual reproduction. Sporangia spores are produced within
spherical sporangia borne on long unbranched sporangiophores
(Fig. 5.1.) The sporangiophore grows as an erect hypha; its apical
growth ceases and the tip swells into a globular structure full of
protoplasm. The spherical structure is separated from the hypha
by a septum which later bulges and projects into the former as a
dome-shaped structure called columella. The spherical structure
including the columella is called the sporangium. The protoplasm
cleaves into numerous multinucleate segments; each of which
secretes a wail and becomes a spore called the sporangiospore.
Meanwhile, the sporangial wall on the external surface develops
minute needle-like crystals of calcium oxalate.
When the sporangial wail dissolves on maturity, the spores remain
firmly adhered to the columella in a drop of fluid and are not blown
off even by strong winds {see Fig. 5.1). The spores when dispersed,
usually by insects, germinate immediately by germ tube or remain
dormant.
Chlamy do spores. Chains of round and thick-walled chlamydos-
pores are formed on the hyphae. When the walls are thin these are
called gemmae. When rest of the hyphae die, the chiamydospores
and gemmae survive.
Torula condition. The hyphae of M. racemosus form yeast like
‘sprout’ cells or oidia in sugar solutions. This is called yeast or
Torula condition’.
Sexual reproduction. It occurs through fusion of similar gametan-
gia (gametangial copulation) and subsequent production of a thick-
walled zygospore. Both homothallic and heterothallic species are
known. The phenomenon of ‘Heterothallism’ was first discovered in
Mucor in 1904 by A.E. Blakeslee (Details see in Chapter 9). The op¬
posite mating types are designated as ‘plus’ (+■) ‘minus (—) strains
which are morphologically alike and hence the terms male and female
could not be used.
Differentiation of sex organs starts when hyphae of similar (in
homothallic strains) or of opposite strains (in heterothallic strains)
come in contact. Gametangial branches, called progametangia, arise
from each of these hypae. Each progametangium enlarges at the tip
which is delimited by a cross wall. The terminal multinucleate swol¬
len structure is called the gametangium. The remainder of the pro¬
gametangium is called suspensor. The two gametangia are of equal
size. The wall between them dissolves and the protoplasm of
sub-division Zygomycotina
s' / If Spores
^ Sporangial || cemented
wall dissolves to columella
Progametangium Sporarigiospores
disseminated ^O®
by termites, ©
Columella
Sporangiophore
Germination
Somatic hypha
/ Cf Mx
©gr Sporanglospores
/ ■
—Germ sporangium
Progametangium M ^
Zygdspore germination
lgiurn^ ^
Gametangium ^
jr Karyogamy 8*
prM'eiosis
Zygospore
\// yz Suspensor>
Jj Gametangial
jff copulation
7 (Plasmogamy)
genus RHIZOPUS
arch over and root like rhizoids that enter the substratum. Sporangio¬
phores arise from the junction of stolon and rhizoids. There is no
such distinction of hyphae in the mycelium of Mucor.
There is difference in the
method of dispersal of spores Dry spores’
also. The wall of the sporan¬
gium of Rhizopus cracks into a
number of small fragments; the
columella collapses .and the
sporangiospores are quickly
blown away by wind.
The sexual reproduction,
though similar to Mucor, differs
in cytological details. Karyo-
gamy takes place before the
onset of dormancy of zygospore,
the meiosis is delayed until its
germination. In Mucor, meiosis Fig. 5.2. Rhizopus: hyphae and
precedes the dormancy. The sporangium.
differences between Mucor and
Rhizopus are given below:
Mucor Rhizopus
family P1JLOBOLACEAE
genus PILOBOLUS
("shot gun’ fungus or the ‘hat-thrower.’)
tight Spores
Sporangium
X/.Drop of....
fluid
®J§B>
Subsporangia!
5 Vesicle ®c &
? Exuded droplets,
°-y7Exuded dropletsjj Gametangium Zygospore
/ Sporangiophore B / /
Suspensor
Young ~~
progametangium
- Trophocyst
about 15 days but usually decline after 7 days. Dung samples should
not be placed in air tight containers as in that condition nematodes
and insects break up the dung and produce anaerobic conditions.
Tliallus. The hyphae are coenocytic, and much branched; charac¬
teristic sporangiophores arise from the hyphae.
Reproduction
Asexual reproduction. Sporangiophores may be terminal or
intercalary in position on the mycelium. If it arises terminally, the
sub-division Zygomycotina 55
order ENTOMOPHTHORALES
genus ENTOMOPHTHORA
Reproduction
Asexual reproduction. The asexual reproduction occurs by:
(1) Budding or fission—Fission is characteristic of bacteria and
among fungi it occurs only in fission yeasts,
(2) Fragmentation, occurs frequently in all fungi,
(3) Chlamydospores, and
(4) Conidia.
The Ascomycotina have specialized in conidial production and it is
said that conidia have reached their zenith in this group. A variety
of conidia are produced in hundred different ways. Conidiophores
may lie externally on the hyphae or inside fruiting bodies (Fig. 6.1)
called pycnidium (pl.~ pycnidia) or acervulus (pl. = acervuli).
The conidiophores may be free or aggregated in various ways. When
the conidiophores are fused it is called synnema (pi. —synnemata).
Less compact fusion is called a coremium. Sometimes,conidiophores
arise from a cushion—like stroma—(a mass of hyphal tissue). It is
called a sporodochium. A pycnidium is a globular or flask-shaped
fruiting body having an ostiole at the top while the acervulus is a
disc-like structure usually formed under the epidermis or cuticle of
the host plant. The conidia are of various shapes, sizes and colours.
Sometimes the conidial masses give characteristic colour to the fun¬
gus e.g., green mold—Penicillium or black mold—Aspergillus.
sub-division Ascomycotina 59
Development of Ascus
The male nuclei, on reaching the ascogonium (Fig. 6.2) form
dikaryons (pairs of nuclei) with the female nuclei. They don’t fuse
immediately to form diploid nuclei as occurs in the Mastigo—and
Zygomycotina. Thus, the karygamy is delayed. A dikaryophase
intervenes between plasmogamy and karyogamy. Small protuberances
appear on the ascognium (or the somatic cell acting as the female
cell), which grow and form ascogenous hyphae. The dikaryons
multiply by conjugate (simultaneous) divisions and the daughter
nuclear pairs migrate into the ascogenous hyphae- Later,septa are:
laid down in the ascogenous hyphae. Each cell of the ascogenous
hypha is dikaryotic except the terminal cell which contains a single:
nucleus. The dikaryophase in Ascomycotina is represented by as¬
cogenous hyphae; rest of the hyphae remain monokaryotic and take
part in the formation of a thick protective covering around the asci-
Any dikaryotic cell of the ascogenous hyphae elongates and bends to
form a hook or crozier-like cell, called ‘crook cell.’ Its two nuclei
divide mitotically in a way that their spindles lie vertically and
parallel to each other (see Fig. 6.2). Two daughter nuclei, one from
each spindle, lie at the bend of the hook while one nucleus lies at the
tip and one near the basal septum. Two septa are laid down to form
three cells—uninucleate basal and apical cells and a binueleate
penultimate cell. The penultimate cell acts as the ascus initial cell.
The basal and apical cells later fuse and the nucleus of the basal cell
migrates into the apical cell, through a pore formed at the point of
contact. Thus, the apical cell becomes binueleate and repeats the
above events to form a second ascus initial cell. This occurs repeated¬
ly, and several penultimate cells are formed which give rise to asci.
The penultimate, or ascus initial cells,elongate into a club-shaped
structure. The two nuclei, which remained as a dikaryotic pair from
the time of plasmogamy, fuse to form a diploid nucleus. Thus, the
Trichogynd Ascogenous hyphae
;sub-division Ascomycotina
Formation of Ascocarps
During the development of asci, the surrounding monokaryotic
hyphae organize a thick protective coat around the developing asci.
The wall is formed by prosenchymatous or pseudoparenchymatous
fungal tissues. The protective coat and the asci are jointly called
as the ascocarp. Three types of ascocarps are known viz., cleisto-
thecium, perithecium and apothecium (Fig. 6.4). Cleistothecium,
sub-division Ascomycotina 63
Cleistothecium
Apothecium
Fig. 6.4. Types of ascocarps. Whole fruiting bodies and their vertical
sections.
V B
V operculum (lid) or by
breaking of the apical
wall. In a large num¬
Fig. 6.5. Types of ascus openings : A. no pores
ber of Ascomycotina
B. apical pore, C. operculum, D. slit, E. bitu¬ the ascospores are re¬
nicate ascus, the inner wall extends out of the leased forcibly.
outer rigid wall. The ascospores when
reach a suitable substratum, germinate and give rise to a monokar
yotic mycelium.
Important Exceptions
1. Ascogenous hyphae, asci and ascocarps are not formed in class
Hemiascomycetes.
2. Asci are formed sometimes in cavities or locules formed in a
stromatic mass of hyphae—it occurs in class Loculoascomycetes.
Classification
The sub-division Ascomycotina is divided into six classes on the
basis of the presence or absence of ascocarp and its shape. The
classes are : (/) Hemiascomycetes, (i7) Loculoascomycetes, (Mi) Plec-
tomycetes, (iv) Laboulbeniomycetes, (v) Pyrenomycetes, and (vi) Dis-
comycetes.
The characteristics of the various classes are as follows:
1. Hemiascomycetes—Asci naked; no ascogenous hyphae, no
ascocarp.
sub-division Ascomycotina 65
class HEMIASCOMYCETES
order ENDOMYCETALES
SACCHAROMYCES CEREVISIAE
(Brewer’s and Baker’s Yeast)
Man has known bread and wine since antiquity but not Saccharo-
myces cerevisiae. However, fermentation, the conversion of sugar
into alcohol, attracted the attention of best minds in the latter half of
the 19th century. This naturally gave a place of pride to yeast.
Louis Pasteur the great scientist of France contended that fermen¬
tation was brought about by the life activities of yeast which grew
and increased in dry weight in sugar solutions. “No fermentation
without life,” claimed Pasteur. Leibig, a German chemist, chal¬
lenged Pasteur’s findings and claimed that the ferment was an extra¬
ordinarily labile organic substance formed by interaction of air with
something present in plant juices. After two years of Pasteur’s death,
Buchner in 1897 produced cell-free yeast extract which brought about
the fermentation. The extract, though derived from yeast, was life¬
less; something like cell-free extract of brain capable of thinking!
Later, the name zymase was given to the chemical that caused fer¬
mentation. It was an enzyme, which literally means ‘in yeast.’ This
discovery of enzyme gave birth to the science of Biochemistry. So
Biochemistry owes as much to man’s thirst for alcohol as Chemistry
owes to his greed of making gold (Alchemy).
Yeasts, because of their ability to ferment sugar, are named Saccha-
romyces (sugar fungi). In, a well-aerated sugar solution the yeast
^ grows and undergoes normal aerobic respiration producing carbon
dioxide and water. However, in absence of oxygen, the aerobic
respiration ceases and fermentation takes place resulting in the pro¬
duction of ethyl alcohol and carbon dioxide. In brewing industry,
alcohol is the desired product,whereas in baker es, C02 is utilized to
make the bread porous and light.
Yeasts are the richest source of Vitamin-B Complex and dried yeasts
in form of tablets are used as a nutritional supplement in medical
therapy.
Habitat. Yeasts occur abundantly on substrates rich in sugars
like the surface of fruits and nectar of flowers etc.
Thallus. Yeasts are unicellular organisms of all possible shapes—
circular, oval, elongated, rectangular dumb-bell-shaped and triangu¬
lar. Individually, the cells are colourless but colonies appear white,
.cream-coloured or light brown. The colony characteristics, along
with physiological reactions, are used in the identification of species.
The colonies of yeasts strongly resemble bacterial colonies.
sub-division Ascomycotina 67
The yeast cell (Fig. 6.6) has a cell wall which consists of mannan
and glucan in addition to chitin. Inside, there is a big vacuole which
occupies a large portion of the cell. The nucleus lies on one side of the
vacuole and very close to it. The cytoplasm, in addition to the diffe-
rent usual cell organelles, contains glycogen, oil and refractile volu¬
tin granules (an inorganic metaphosphate polymer) as reserve mate¬
rials. The vacuole was previously considered to be a part of the nu¬
cleus but the electron microphotographs have conclusively demons¬
trated that the nucleus and the vacuole are separate entities, each
surrounded by its own membrane.
Life History
S. eerevisiae shows an alternation of haploid and diploid genera¬
tions (Fig. 6.7) which is a rare thing in fungi (another example is
Allomyces, an aquatic Chytridiomycete). Each generation is perpetu¬
ated by budding and is equally extensive and important.
In budding, a small protuberance appears at one or more places on
the cells, which grow into daughter cells. The nucleus divides by
constriction; the nuclear envelope does not break. Chromosomes are
not seen even in electron microphotographs. The daughter nucleus
enters the bud along with the other organelles. Before the daughter
cell is pinched off by formation of a wall, it produces another cell
68 A Textbook of Fungi, Bacteria and Viruses
The haploid cells of (+) and (-) mating types, grow indepen¬
dently and multiply by budding. After a period of growth two haploid
cells of opposite strains copulate to form a diploid cell. The diploid
yeast cells are bigger and their colonies grow faster. The diploid
sub-division Ascomycotina 69
cells also increase their number by budding until the conditions* for
a change to haplophase arise. The diploid cells act as asci. The
diploid nucleus undergoes meiosis and four haploid nuclei, two of
each strain, are produced. Ascospores are formed incorporating
these nuclei. The ascospores are liberated by breaking of the ascus
wall. Each ascospore resembles an yeast cell though much smaller.
They grow into haploid cells and start budding to establish the haploid
phase, thus completing the life cycle.
class PLECTOMYCETES
♦In laboratory the yeast cells can be induced to form ascospores by growing
them on agar containing 0.5% sodium acetate but no other nutrients. *
70 A Textbook of FungU Bacteria and Viruses
Reproduction
The conidial stage is dominant and the sexual state is either absent
or rarely produced. Species which have only conidial stage are
called Aspergillus while those in which sex organs are formed are
designated as Eurotium.
Asexual reproduction. Conidia are produced in enormous
quantity. The mycelium starts forming conidiophores very early.
The long and erect conidiophores. arise from a particular thick-walled
foot cell. (Fig. 6.10). It forms a terminal swollen vesicle. Bottle
shaped structures called sterigmata or phialides, arise all over the
surface of the vesicle and produce chains of conidia at their tips.
When there are two layers of sterigmata, the lower most is called
primary sterigmata or metulae and the upper, as secondary sterigmata
or phialides. The phialides are bottle-shaped. The neck is called
the spore-producing tube; the conidia are formed inside this tube
(Fig. 6.9). A portion of protoplasm with a nucleus,inside the neck, is
sub-division Ascomycotina 71
Aseogonium
Antheridium
Anthericfruro.
74 A Textbook of Fungi, Bacteria and Viruses
genus PENICILLIUM
Reproduction
Asexual reproduction. The dominance of the conidial state in
the life of Penicillium is greater than even in Aspergillus. The chara¬
cteristic conidial apparatus (Fig. 6.11), which resembles a brush or
broom, is called a Penicillus (Penicillium = small brush). Long,
septate conidiophores
arise from any cell of Conidia
the hypha, and not
from a foot cell as in
Aspergillus. The coni¬
diophores branch
once or twice at two-
third of its total length.
These branches are
called primary sterig-
mata (^rammi) and
secondary sterigmata
{=metulae), which
finally bear the bottle¬
shaped phialides.
Fig. 6.11. Types of conidiophores of Penicillium,
Conidia (phialospores) A. P. thomii, B. P. cyclopium.
are produced in the
neck region of the phialide,as described earlier for Aspergillus. Long
chains of conidia are formed with youngest conidium at the base.
These are globose to ovoid and look like glass beads under the micro¬
scope. As in Aspergillus, the coloured conidia—green, blue, or yellow,
give the characteristic colour to the colony, which aid in identification
of species. In some species,the conidiophores unite to form coremia
(sing, coremium).
Sexual reproduction. In most of the species sexual reproduction
does not occur at all. Conidia are the only method of reproduction.
Even the species in which sexual reproduction is known, it is a rare
event and, therefore, of academic importance only.
The uninucleate mycelium forms a swollen cylindrical ascogonium
(Fig. 6.12) which is uninucleate in the beginning but by repeated
nuclear divisions comes to have 64 nuclei. An antheridial branch,
which originates on a separate hypha, coils around the ascogonium
and cuts a terminal antheridium. The wall at the place of contact
of the two sex organs dissolves but nuclear migration from the
antheridium has never been observed. The male nuclei can be seen
in the antheridium even after the formation of ascogenous hyphae.
A Text took ofFungi, Bacteria and Viruses
class PYRENOMYCETES
order ERYSIPHALES
family ERYSIPHACEAE
Powdery Mildews
The ‘powdery mildews’ are obligate parasites of higher plants mostly
dicotyledons. There are eight genera (Yarwood, 1973). These are :
Sphaerotheca, Podosphaera, Erysiphe, Phyllactinia, Microsphaeray
Uncinula, LeviHula and Acrosporium. The genera are identified on
the basis , of (0 the nutnber of asci (one or many), and (//) the
appendages present on the cleistothecium.
Economic Importance
The powdery mildews, except a few, like the Powdery mildew of
grape, generally do not cause much damage, and are easily controlled.
Some important powdery mildew diseases are the following:
Powdery mildew of grapes— Uncinula necator
Powdery mildew of rose —Sphaerotheca pannosa
Powdery mildew of wheat —Erysiphe graminis
Powdery mildew of ‘Shisham’—Phyllactinia corylea
(Delbergia sissoo)
UNCINULA NECATOR
(Powdery mildew of Grapes)
Reproduction
Asexual reproduction. Large number of short, erect, hyaline
conidiophores arise from the mycelium and cut a row of oval, hyaline
conidia. A conidiophore has a stipe of one or more cells, a generative
cell and one or more maturing conidia. The enormous number of
conidia form a powdery coating on the leaves- hence the name
powdery mildew. The conidia have a high water content and need
no free-water for germination. Maximum germination occurs in
remarkably low humid atmosphere. It is interesting that free water is
inhibitory for germination and sometimes lethal to the conidia.
The conidia, largest in fungi (20-50/m),. are wind-disseminated. On
reaching a suitable host, these germinate and form hyphae which lie
externally on the leaf surface. The hyphae form haustoria which
penetrate only the epidermal cells and derive nutrition without caus¬
ing any visible damage to the host cells.
Sexual reproduction. Coloured cleistothecia appear on the super¬
ficial mycelium at the end of the host season. Sex organs are borne on
adjacent hyphae. A globular, single-celled and uninucleate ascogon¬
ium is formed by swelling of one of the cells. The antheridium, which
is also uninucleate, is stalked and is thinner than the ascogonium.
Plasmogamy occurs when the intervening walls dissolve. Pairing of
nuclei takes place. The ascogonium elongates and becomes 2-3 sep¬
tate. Ascogenous hyphae arise from the central cell and form asci
without formation of croziers. The ascus formation is accompanied
by the development of the peridium from the surrounding somatic
hyphae. The peridium is 2-3 layers thick; the inner most layer consist¬
ing of thinner nurse cells (used up by developing asci). Appendages
with coiled tips, a characteristic feature of Uncinula, arise from the
outer most peridial layer. Appendages with coiled tips and the many
asci are the two diagnostic features of this genus.
Cleistothecia are minute, round and black structures,, which repre¬
sent the resting stage of the* fungus. At the return of suitable
weather, the ascocarp breaks by the pressure generated due to swel¬
ling of asci after absorbing moisture. The asci spread out like a fan,
and finally, when the ascus wall bursts, the ascospores are liberated.
The ascospores germinate only on the host surface to produce the
superficial mycelium.
In India, the cleistothecia of Uncinula have not been reported and
80 A Textbook of Fungi, Bacteria and Viruses
Conidia
Conidium''\1 Appresorium
W'/:f—Conidiunr.
Infection
Generative hypha
cel!
Conidiophore
Stipe Epidermis of
Hypha
Haustorium host leaf
Ascogonium
Antheridtum
Appresorium
Germination on
host epidermis
Gametangia!
contact
(Plasmogamy)
Ascogenous
hyphae
Appendages
Cleistotbecium^Ss^ -21
Fig. 6.13. Life cycle of Uncinula necator causing Powdery mildew of grapes.
sub-division Ascomycotina
81
therefore, the recurrence of the disease is supposed to be through
conidi a only.
order SPHAERIALES
jgenus CHAETOMIUM
ASCOSPOTFS
. Ascus Extruded ascospores
-Curly appendages
Mycelial appendages
Reproduction
Asexual reproduction is rare. Only five out of over 85 species
are reported to produce conidia. The perithecium, a product of sexual
reproduction, is flask-shaped having an elongated beak which has a
distinct ostiole. The perithecium is covered by long hairs. These are
of two types. Those at the neck are curly while those at the base are
straight. The ascospores lie free in the perithecial cavity in a gelati¬
nous mass formed by dissolution of the ascus walls. The ascospores
are always unicellular and dark. They germinate to form the myce¬
lium which later produces the perithecia.
class DISCOMYCETES
order PEZIZALES
genus PEZIZA
Reproduction
Asexual reproduction. It is rare. However, in P. repanda and P.
vesiculosa conidia are reported to be formed. It’s role in the life cycle
is negligible.
Sexual reproduction. Somatogamy between hyphae of opposite
strains (in heterothallic species) or between two cells of the same
hypha (in homothallic species), results in plasmogamy and formation
of a dikaryotic cell. Sexual organs are not formed at all. Ascogenous
hyphae arise from the dikaryotic cell and the fruiting body is organiz¬
ed around the developing asci (Fig. 6.16). Eventually, a cup-shaped
apothecium is formed.
The apothecium consists of three parts (Fig. 6.17): (/) the hymen-
ium, (/'O the hypothecium, and (///) the excipulum. The hymenium,
which consists of a layer of asci and paraphyses, lies at the inner sur¬
face of the cup. The asci are club-shaped or cylindrical having eight,
obliquely-arranged ascospores. The paraphyses, which are found
intermingled with the asci, may be longer, equal or shorter than the
asci. Sometimes, the tips of the paraphyses branch, which unite to
form a layer called epithecium, covering the hymenium. The
hypothecium is a thin layer of interwoven hyphae lying below the
hymenium. The main body of the apothecium is called excipulum.
84 A Textbook of Fungi, Bacteria and Viruses
Hymenium
Subhymenium
Excipulum
Apothecium
Substratum
— C
Pseudoparenchymatous
tissue of ectai excipulum
Prosenchymatous tissue
of medullary excipulum
genus MORCHELLA
(The Morels or Sponge Mushrooms)
The morels are extremely delicious to eat and are popular in India by
the name ‘Gucchi.’ The various true morels are the different species
of Morchella viz., M. conica (conical morel), M. delicosa (delicious
morel), M. hybrida (hybrid morel), M. esculenta (the common morel).
All are edible. Unfortunately, they have not been cultivated com¬
mercially. Their fruiting is uncertain and we don’t know the condi¬
tions that induce fruiting in the vegetative thallus. Even in labo¬
ratory, we grow them only in the mycelial stage; fruiting never occurs
in laboratory pure cultures. This is the reason the morels continue
to be the food of the rich.
Habitat. Like Peziza, species of Morchella grow on manure heaps,
humus-rich soils, old logs and stumps in forests.
The Thallus. The mycelium grows inside the substratum and
derives nutrition from the organic material of the substrate. The
fruiting bodies, the visible edible part, are formed especially after
rains.
The fruiting body. It is a stalked
apothecium, 10-15 cm in height
(Fig. 6.18). The thick stalk stipe is
hollow and bears a sponge-like or ‘honey
comb’ like pitted pileus. Both stalk and
the pileus are hollow. The pileus is
whitish grey to dark brown, depending
on the species and the age. The pileus
has convoluted ridges and furrows
(Fig. 6.19). The furrows are lined by
hymenium consisting of asci and para-
physes. The asci have curved tips and
contain 8 ascospores. The ridges lack the
Fig. 6.18. Morchella
hymenium. The ascospores are violently esculenta, the * fruiting
discharged. body.
Life cycle. The ascospores give rise to haploid monokaryotic
mycelia. Somatogamy between cells of opposite strains results in
the formation of a dikaryotic cell. Ascogenous hyphae originate
from this dikaryotic cell which later form the asci. The fruiting
body is formed by the vegetative hyphae and, ultimately, the stalked
apothecium is formed.
86 A Textbook of Fungi, Bacteria and Viruses
The mushrooms and toad stools with which mycology Started, the
rusts and smuts, which have been devastating crops since recorded
history, the ‘shelP and ‘bracket’ fungi that lie attached to the trees
sapping their vitality, all belong to Basidiomycotina. These are the
highest evolved fungi.
The important characteristics of the sub-division are:
1. The basidium,
2. The extensive dikaryophase,
3. The clamp connections, and
4. The dolipore septum.
The Basidium
Like the ascus of Ascomycotina, the basidium is a diagnostic feature
of Basidiomycotina. A typical basidium is a club-shaped structure
bearing apically four basidiospores on pointed projections called
sterigmata. The basidium may be septate, phragmobasidium or
unseptate, holobasidium. In rusts and smuts, the basidia are entirely
of different nature. Basidiospores are formed after karyogamy
and meiosis of the diploid nucleus in the basidium, and are, therefore,
uninucleate and haploid. These may be of two or more mating
types. On germination, the basidiospores give rise to monokaryotic
primary • mycelium.
The Thallus. Three types of mycelia viz., primary, secondary and
tertiary are found in the Basidiomycotina.
The primary mycelium is monokaryotic (i.e., made of haploid
uninucleate cells) and is formed by germination of the basidiospores.
These are of different mating types. It exists for a very short period
and gives rise to the dikaryotic secondary mycelium.
The secondary mycelium. It consists of dikaryotic (binucleate)
hyphae formed by the following methods:
1. somatogamy between cells of monokaryotic hyphae,
2. fusion of two basidiospores, and
3. spermatization of female sex organs.
The single binucleate cell, formed by fusion of two cells of the
primary mycelia or between two basidiospores, divide repeatedly and
88 ,
A Textbook of Fungi Bacteria and Viruses
Clamp Connection
It is a hook-like structure associated with the conjugate division of
the dikaryons in the secondary mycelium. It is a by-pass for the
nuclei, if they cannot pass through the septal pore, as may happen
due to presence of a dolipore septum. A small outgrowth called
‘clamp-connection’ arises between the two nuclei (a and b) of a
binucleate cell (Fig. 7.1) and forms a curved hook. The two nuclei
divide simultaneously. Their spindles are oriented in such a way
that the daughter nucleus of the upper nucleus lies in the clamp while
that of the lower nucleus comes close to the upper nucleus. The
clamp bends and its tip touches the cell; the intervening walls dis¬
solve and the nucleus in the clamp migrates into the cell to lie close
to the lower nucleus. Two septa are now laid down: one longitudi¬
nal at the place of the origin of the cl&mp connection, so as to cut
its original connection wi(h the cell, and the other transversely at
right angles to the first longitudinal septum. A new daughter cel! is;
sub-division Basidiomycotina 89
Clamp connection
/y\
Daughter
cell
i
i
Fig. 7.1. Cell division with clamp connection formation.
Basidiospore
Basidium
\ Karyogamy
Us t/la go
simultaneously.
.Discharged
[ basidiospore
Basidiospore
bubble Bubble
Collapsed:
sterigma
Classification
The sub-division Basidiomycotina consists of 3 class-es: the Telio-
mycetes, the Hymenomycetes, and the Gasteromycetes.
class TELIOMYCETES
order UREDINALES
1. The order includes the rust fungi which total about 4000 species
distributed among 100 genera. All are obligate parasites of angio-
sperms, gymnosperms and ferns. Some of the rust diseases do great
'damage to our crops e.g., wheat rust, coffee rust etc.
2. Basidiocarp is absent and,therefore,there is no tertiary mycel¬
ium.
3. The. hyphae—usually secondary mycelium, remain in the inter¬
cellular spaces of the host tissue and derive nutrition through haus-
toria.
4. Clamp connections are rare.
5. Teliospores originate from the apical cells of dikaryotic hyphae.
These may be unicellular or multicellular. The structure of teliospores
forms the basis of identification of the rust genera.
6. The rusts have a pleomorphic life cycle, which in its basic form
contains five distinct spore-states,that differ morphologically and cyto-
logically. These are pycnia, aecia, uredinia, telia and basidia.*
If all the states are produced on the same host the fungus is called
•Various synonyms exist for each of these stages: Pycnium (syn. spermo-
£onium, pycnidium), Aecium (syn.' Aecidium, cluster cup), Uredinium (syn.
uredosorus, uredium), Telium (syn. teliosorus, teleutosorus).
94 A Textbook of Fungi, Bacteria and Viruses
autoecious. But in many rusts these spore states are formed on two
hosts, the pycnia and aecia occurring on one host and the uredinia
and telia, on the other. Such rusts are called heteroecious. On
germination the teliospores form basidiospores.
All the spore forms may not be formed in the life cycle of a rust,,
and based on this, three types of life cycles are recognized—the
macrocyclic, demicyclic and microcyclic. In the macrocyclic or
long-cycled life cycle, all the five spores are formed. In demicyclic
rusts uredinia are not produced. Microcyclic life cycles have only telia.
The macrocyclic and demicyclic rusts may be autoecious or heteroe¬
cious,but the microcyclic rusts are, by necessity, always autoecious.
The pycnia represent the gametic stage; the aecia represent the
stage in which dikaryotization occurs; the uredinia represent the
conidial or repeating, asexual stage, while the telia and basidia
(=promycelia) represent the stages in which occur the karyogamy
and meiosis. The term spermogonium is frequently used for the
pycnium. But this is not correct, as spermogonia denotes only ‘male’
sex organs while rust pycnia bear both male and female structures
(the spermatia and the receptive hyphae).
The order Uredinales includes 3 families: Pucciniaceae, Melamp-
soraceae and Coleosporiaceae (Wilson and Henderson, 1966). In
Pucciniaceae the teliospores are stalked while in the other two
families they are sessile. We shall study the genus Puccinia belonging
to the family Pucciniaceae.
genus PUCCINIA
WHEAT RUST
Three types of wheat rusts are known and all of them occur frequent¬
ly in India. These are:
(/) Black or stem rust caused by P. graminispritici
07) Orange or brown rust caused by P. recondita (=syn. P. triti¬
cina).
(iii) Yellow or stripe rust caused by P. striiformis (-syn. P. glu-
maruni).
Fig. 7.5. Uredial and telial stages of Puccinia gar minis tritici.
on wheat leaf.
96 A Textbook of Fungi, Bacteria and Viruses
Cluster cups(Aecia)
*In thistle rust, caused by P. obtegens, the pycnia emit a sweet smell like
that of violets. It is claimed that a scent is associated with Barberis leaves bear¬
ing pycnia of P. graminis,but it is too faint to be detected unless one has a very
sensitive nose.
Promycefium
+_ Basidiospores of
p»ir ✓ different strains
Meiosis m 1
Basidiospores infect
barbery leaf and from
m- pycnia of opposite strains.
Teliospore germination ‘
/I Teliospore
y\ i
Nectar drop
Spermatiumof opposite
strain fuses with
receptive hypha
pycnium
Reinfection of
Uredospore wheat leaves through
x stomata
rr\ /£\_Aeciospores
r*
sub-division Basidiomycotina 99
The uredial and telial stages of the brown and yellow rust of wheat
are depicted in Fig. 7.8. The symptoms and important differences
between the three rusts of wheat are given in Table 7.1.
Brown rust
Fig. 7.8. Uredial and telial stages of the brown and yellow rust of wheat.
Table 7.1. Comparison of the three rusts of Wheat
Black rust (P. graminis) Orange or brown rust P. recondita Yellow rust P. striiformis
(=P. triticina) glumarum)
. Stems are most seriously attacked, followed Attacks leaves almost exclusively, very Leaves are most severely attacked followed
by leaf sheaths, leaves and ‘ears’, in decreas- rarely leaf sheaths or stem. by leaf sheaths, stems and eats.
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►. 240 races of the pathogen are known; 14 re- 100 races are known but only 14 are re- Out of the 14 races, 10 are reported from
ported from India. ported from India. India.
. The annual recurrence of the disease on the Same as of black rust. In addition to uredospores from hillsvthe in¬
plains is through uredospores blown down oculum initiating primary infection is provid-
from hills late in the wheat season. ed by some locally growing “collateral hosts.”
102 A Textbook of Fungi, Bacteria and Viruses
Physiological Races
P. graminis infects several hosts belonging to the family Graminae.
The uredospores of P. graminis, which infect wheat, fail to infect
another host-say oat or barley. Similarly, uredospore produced
by P. graminis on barley, fails to infect any other host. Erikson,
(1891) a Swedish botanist, designated each isolate by a third name
(trinomial nomenclature) and called it sub-species or “formae
specialis.” Thus, the name P. graminis tritici was given to the
isolate that infects wheat; P. graminis hordei for the isolate infecting
barley and P. graminis avenae for the isolate infecting oat. Laterrit
was found by Stakman and Levine of USA (1922) that in P. graminis
tritici, the-isolate that infects one variety of wheat fails to infect ano¬
ther variety. Thus, a further identification is needed to indicate the
variety of the host. These are called physiological races. Over 200
physiological races of P. graminis tritici are known. The physiological
races are designated by numbers e.g., P. graminis tritici 138. The
work of identification of the physiological races is very important.
Before developing a resistant variety a knowledge of the existing
races is essential. It is a difficult job and is done only by specialists.
But we can see how it is done. For delineating the races of Puccinia
graminis tritici, uredospores, obtained from different areas, are ino¬
culated on ‘differential varieties’ (these are 12 in number for wheat
rust fungus). The infections produced, on the basis of number and
size of pustules formed, are graded on a standard chart as follows:
0. Immune—no pustules produced, no sign of infection.
1. Very resistant—only faint chlorotic spots surrounded by necro¬
tic tissues appear.
2. Moderately resistant—minute scattered pustules appear.
3. Moderately susceptible—medium size pustules and chlorotic
spots appear.
4. Very susceptible—medium to big size confluent pustules formed.
X. Heterogeneous—size of pustules vary but mainly mixture of
type 1 and 4.
Small variations in the grades are indicated by one or two plus or
minus signs e.g. 2~, 2""", 2+, 2++ etc. The identification of physiologi¬
cal races is a difficult task; the fluctuating effect of the environmental
factors adds to the complexity. Thus, certain races produce the X-
reaction under some conditions but not under others.
Fungi are a treacherous lot. Their physiologic races can be further
demarcated into “biotypes” by adding another variety to the stan¬
dard differentials. Uppal and Gokhale (1947), Gokhale (1950) and
:sub-division Basidiomycotina 103
order USTILAGINALES
1, The fungi included in this order are called smut fungi and the
diseases they cause are called smut diseases.
2. The teliospores are formed from all the cells of the secondary
dikaryotic mycelium by developing a thick resistant wall.
3. Basidiospores are formed directly on a septate or non-septate
promycelium produced by the teliospores on germination. There are
no sterigmata. The basidiospores are passively discharged.
4. Except one small family—Graphiolaceae, smuts do not form a
basidiocarp.
5. Mycelium is intercellular and forms haustoria which draw7 nutri¬
tion from the hosts cells.
6. Clamp connections and dolipore septa absent (Talbot, 1970).
The Ustilaginales and Uredinales show close resemblance. Both
groups are important plant pathogens growing in the intercellular
spaces and deriving nutrition through haustoria. Both lack dolipore
septa and clamp connections. The differences between them are
given in the table below.
genus USTILAGO
Fig. 7.14- A 'fairy ring' of Agaricus in natural Fig. 7.15. Spore print of
habitat. Agaricus.
Fig. 7.19. Amanita phalloides—\he Fig. 7.21. Pofyporus betulinus.
fruiting body in natural habitat. natural view.
life
sub-division Basidiomycotina 105
Reproduction
Asexual reproduction. Budding of basidiospores is the most
common method of asexual reproduction. Fragmentation, and in
some species,conidia are the other means of asexual reproduction.
Sexual reproduction. Sex organs are absent. But the sexual
reproduction, represented by karyogamy followed by meiosis, does
occur and brings about the genetic recombination. The function of
sex organs is taken up by somatic cells which transform into telio¬
spores. Plasmogamy, karyogamy and meiosis are separated in space
and time, i.e., occur at different places and at different time. Plasmo¬
gamy is brought about by the fusion between two uninucleate cells
of opposite mating types, which may be the basidiospores or cells of
the primary mycelia. Most of the smut fungi are heterothallic. The
karyogamy is delayed until teliospores germinate and form a pro¬
mycelium. The diploid zygote nucleus undergoes meiosis to form
four haploid nuclei, two of each mating type.
The promycelium is divided into four cells by horizontal septa.
A bud arises,from each cell into which one nucleus passes. The bud
develops into a basidiospore and is pinched off. In some species the
promycelium may continue forming the basidiospores. In some
species e.g. U. maydis, the basidiospores themselves by budding form
sprout cells ( = daughter basidiospores or secondary sporidia). In U.
tritici there are no basidiospores; the cells of the promycelium form
infection threads which fuse to establish a dikaryotic hypha that
grows and develops into the secondary mycelium.
symptoms appear only when ‘ears’ come out. Sometimes the infected
plants are stunted in growth. The ears instead of containing grains
contain black mass of spores (Fig. 7.9). The sori In the early stages
are covered by a thin membrane derived from host tissue but later
the membrane ruptures and the spores become a loose mass; hence
the name ‘loose’ smut.
Life History
The disease is internally seed-borne i.ethe fungus lives in dormant
stage in the embryo of the seed (Fig. 7.10). Such seeds look perfectly
Teliospores blown to
wheat flowers,
Germination
/ on wheat flowers
Karyogamy
Promyceluim
"xmlf/fi
Teliospores I
i _ lx „ f » Smutted ear
formed in ^ f
inflorescence]
Oikaryoti^uTton
infected seed
Systemically
Seed germination
infected plant
normal and there is no method to detect the infection unless they are
grown and watched till the^ar’formation. The seedlings, and later
the whole plant is infected by its extensive dikaryotic mycelium,
which grows intercellularly and derives food through haustoria. The
plants are said to be systemically infected. The hyphae collect in
the ovaries and the hyphal cells found off, develop thick walls and
sub-division Basidiomycotina 107
Control
1. Seeds for sowing should be obtained from reliable places where
the disease does not occur.
2. If there is any doubt, the seeds may be treated by ‘hot water
method’. The method was discovered by Jensen in 1889. Seeds are
soaked in shallow warm water contained in pots and then spread in
hot scorching sun during the day. The hyphae die due to the heat of
the sun, much quicker than the embryo. The safety line is very thin
and there is every chance of killing the seeds. But the risk is worth
taking.
3- Use of resistant varieties is the best method of avoiding the
disease.
108 ,
A Textbook of Fungi Bacteria and Viruses r
SMUT OF MAIZE
Pathogen—Ustilago maydis
The pathogen is much different from the other smut fungi. The
important differences are the following:
1. In addition to parasitic life, it grows well saprophytically in
soil.
2. It grows intracellularly and does not form haustoria.
3. Does not grow systemilcally in the host but remains localized
to several independently infected parts.
4. Smut sori are formed on almost all the above-ground parts of
the plant—the cob, the leaf and the stem.
5. The fungus induces tumor formation, and outgrowths, as big as
a child’s head, are frequently formed. These are filled with telio-
spores.
6. The primary mycelium, which in other species is incapable of
growth in the host tissue, grows sufficiently well in the host tissue.
However, the dikaryotic secondary mycelium grows more vigorously
than the primary mycelium.
Symptoms. Sori (galls) containing teliospore are formed on all
above-ground parts. Galls of different sizes appear; the biggest galls
are formed on the cobs.
Life History
The fungus lives saprophytically in soil. When the maize crop is-
available in the field, the basidiospores, blown by wind currents,
reach the . plants and germinate to form a monokaryotic primary
mycelium (Fig. 7.11). The primary mycelium makes some growth in
the host tissue, but the vigorous growth starts only when the dikaryo¬
tic secondary mycelium is established by anastomosis and fusion of
hyphae of opposite mating types. Dikaryotization is necessary for
gall formation. The fungus does not spread in the host tissue but
remains confined to the vicinity of the place of infection and forms a
sorus of teliospores. Big masses of sori appear in the form of galls
or tumors. Each gall is the product of a separate infection. The telio¬
spores are formed by rounding up of the binucleate cells. In the
beginning the telia are covered by the host tissues but later the
covering ruptures and the teliospores become free. The spores are
disseminated by wind. They germinate to form a club-shaped promy¬
celium in which karyogamy and meiosis occur, resulting in the for¬
mation of four haploid nuclei. Segregation of sex occurs during.
sub-division Basidiomycotina 109
J leiosis
and out of the four nuclei formed, two are of one mating
type and two of the other mating type. Four septa are laid down
Promycelium
■f*
•5 (el's
r. ...4
Teliospore
Teliospores
Sporidia
®/
j -J* Secondary
Dikaryotic / Jsh~ sporidia
secondary hypha
Germination
Monokary otic
primary hypha
Dikaryotization
"Which divide the promycelium into four cells, each containing one
haploid nucleus. A bud develops on each cell into which passes the
nucleus. The bud later develops into a basidiospore. The basidio-
spores are passively discharged, and blown by wind to cause more
infections. These form secondary sporidia by budding.
Control Measures
1. Use of resistant varieties is the safest and cheapest method of
control.
2. Soil sanitation. The soil should be sprayed with fungicides to
kill the fungus present in the soil.
class HYMENOMYCETES
order AGARICALES
The generic name Psalliota has been used for this fungus in older
literature. Agaricus campestris is cultivated extensively under the
industrial name A. bisporus. In culture,the physiological and morpho¬
logical characters show marked differences to justify the change of
name.
Habitat. The fungus grows in lawns, fields and forests round the
year. The extensive mycelium remains hidden in the soil; only the
fruiting body is visible.
Thallus. The mycelium, which remains underground and grows
saprophytically, is the dikaryotic,secondary mycelium. It is formed
by somatogamy between monokaryotic primary mycelia of different
mating types formed by germination of the basidiospores. The pri¬
mary mycelium is of short duration. The fruiting body, which forms
the magnificent umbrella aboveground (Fig. 7.12), is made up of
dikaryotic hyphae called tertiary mycelium. The fruiting body is
ephemeral and lives only for a few days. The secondary mycelium,
however, is perennial and continues growing for several years and
form fruiting bodies year after year. The fruiting body, or the
basidiocarp, (Fig. 7.13) has a stalk (stipe) and a circular cap (pileus)
A skirt-like ring of tissue, called annulus, surrounds the stipe a
little below the pileus. The gills (=lamellae) can be distinctly seen
if the stipe is removed and the pileus is inverted. The pileus on the
under surface, bears numerous, vertically-hanging gills, which con¬
verge from periphery towards the stipe. The gills are of different
lengths and bear basidia all over the surface. The basidia produce
basidiospores in astronomical numbers.
112 A Textbook of Fungi, Bacteria and Viruses
sub-division Basidiomycotina -113
Fairy Rings
The fruiting bodies of Agaricus campestris form “fairy rings?’ Th ^
umbrella-shaped basidiocarps, ‘The fungus flowers”, formed in circles
on the ground, provide a beautiful spectacle which could be described
as fairies dancing in a circle (Fig. 7.14).
The fruiting bodies being ephemeral, the pleasant sight is short¬
lived. But the mycelium persists in the soil and continues growing as
a circular colony, in the same way as a circular fungus colony grows
in a Petri dish in the laboratory. As the outer edge of the colony
grows, older parts in the centre die and degenerate. The next time
when the fruiting bodies are formed the ring is bigger in diameter.
The dead remains of the fruiting bodies make the soil more fertile
and rings of greener grasses are formed where once stood the um¬
brellas.
Reproduction
Asexual reproduction. A. campestris produces only chlamydos-
pores; conidia and oidia are not known.
Sexual reproduction. The sexual reproduction is represented by
karyogamy and meiosis, which occur in basidia, borne on the gills.
The nuclei of opposite mating types come together and form dikar-
yons after the fusion of the monokaryotic primary hyphae. The
dikaryons multiply by conjugate divisions in the extensive, secondary
mycelium. Ultimately, club-shaped basidia develop from the terminal
cells of these secondary hyphae. The binucleate cell enlarges and
becomes broader and club-shaped. Karyogamy and meiosis occur
resulting in the production of four haploid nuclei. Segregation of sex
occurs during the meiosis and the nuclei are of two or more mating
types, depending on the type of the heterothallism (bipolar or tetra-
polar). The nuclei later migrate into the sterigmata and from there
into the basidiospores, perched asymmetrically on the tips of the ster¬
igmata. The basidiospores, on germination, form primary mycelia,
which by anastomosis and plasmogamy,establish the secondary myce¬
lium. The basidiospores, which are borne asymmetrically on sterig¬
mata are discharged forcibly by water drop method {see Fig. 7.4).
The basidiospores are produced in enormous numbers. If the earth
is not covered by mushrooms, this is because the chances of failure
of spore germination are also equally great. Spores “rain down” at
a rate of half a million spores per minute during the two or three
days of the existence of the sporophore (fruiting body). If the pileus
114 A Textbook of Fungi, Bacteria and Viruses
Rqa:j-
Basidiospores
Basidiospores.
.»_o
Fig. 7.17. A. campestris, showing, A. path (trajectory) of
discharged basidiospores, B. the discharge of basidio¬
spores one after the other.
vertically. The distance between the gills far exceeds the distance
to which the basidiospores are shot. Even a little tilt of 5° of the
gjtts from the exact vertical position may obstruct the free down¬
ward fall of the spores. But this is not allowed to happen. The
sub-division Basidiomycotina 117
gills are positively geo,tropic and any deviation from the vertical
position is immediately corrected by growth movements at the place
of attachment of the gills with the pileus. The life cycle of the fung-
gus is depicted in Fig. 7.18.
MUSHROOM CULTIVATION
A B
family POLYPORACEAE
genus POLYFORUS
This is the largest genus of Polyporaceae and its several species are
important wood-rotting fungi. P. sulphurens causes wood rot of
oaks and other trees; P. squamosus causes ‘heart-rot’ of a number
of forest trees; P. betulinus causes ‘heart-rot’ of birch trees
(Fig. 7.21). Destruction of the heart wood makes the trees hollow
which eventually die. Some, e.g., Polyporus schweinitzii, attack the
lower part of the trunk and the roots (butt-end rot), making the
trees liable to snap off in gales.
The mycelium ramifies within the substratum. It consists of sep¬
tate dikaryotic hyphae which usually show clamp connections. The
fruiting body (the basidiocarp or sporophore) develops as a lateral¬
ly-stalked, fan-shaped, bracket which may be 20-40 cms in diameter
and 2-3 cms thick. A vertical section of the fruiting body (Fig. 7.22}
shows the following zones: (0 pileus surface, (it) context, (Hi) tube
sub-division Basidiomycotina 121
class GASTEROMYCETE3
order LYCOPERDALES
Includes puffballs and the earth stars. The fruit body is epigeal
(the earlier stages are often hypogeal), globose, which varies in size
from a few mm to over 1 meter in diameter; usually sessile, rarely a
few have a pseudostem. The peridium is divisible into two layers, the
exoperidium and the endoperidium. The fertile part, called gleba,
consists of smooth, light coloured spores and well-developed capiili-
tium. The order is divided into several families. We shall study
typical genera of families Lycoperdaceae and Geastraceae.
ECONOMIC IMPORTANCE
Several plant diseases (especially leaf ^pots, fruit rots and wilts)
and animal as well as human diseases are caused by these fungL
Some important diseases are given below:
sub-division Deuteromycotina 125
Plant Diseases
Wilt of cotton — Fusarium oxysporum f. sp. vasinfectun.
Wilt of brinjal -Verticillium dahliae
■Grey mold of apple and strawberry fruits—Botrytis cinerea.
Foot rot of rice - Fusarium moniliforme
Blast disease of rice-Firicularia oryzae
Brown leaf spot of rice—Helminthosporium oryzae
Early blight of potato —Alternaria solani
Leaf spot (Tikka disease) of groundnut - Cercospora personata
Grey blight of tea leaves—Pestalotiopsis theae, (—Pestalotia theae)
Human Diseases
Head “ring worm”—Microsporum audouini
Foot “ring worm” or 'Atheletes foot’—Trichophyton interdigitale
Animal Diseases
Head ring worm of cats and dogs -Microsporum canis
CLASSIFICATION
Earlier these fungi were grouped by Saccardo (1899) into four orders,
Moniliales, Sphaeropsidales, Melanconiales and Mycelia-Sterilia. In
Moniliales, the conidia. and the conidiophores are borne directly on
hyphae. In Sphaeropsidales, the conidia are formed inside flask¬
shaped pycnidia,and in the , Melanconiales, the conidia are formed
in flat,disc-shaped structures, called acervuli, formed beneath the
epidermis or cuticle. In Mycelia Sterilia, as the name itself suggests,
the fungi are sterile and do not form conidia at all, but do form
sclerotia which serve as propagules. In the classification followed
here (Ainsworth, 1973),the Moniliales and the Mycelia Sterilia are put
together under the class Hyphomycetes, while Sphaeropsidales and
Melanconiales are merged to form the class Coelomycetes. The ‘false
yeasts’, (that do not form ascospores), which were earlier placed
under Moniliales,are now treated as a separate class-Blastomycetes.
class HYPHOMYCETES
The conidia and the conidiophores are borne directly on the hyp-
hae. Some of the genera have mycelium which is sterile i.e., do not
form spores. On the basis of conidial ontogeny i.e., the mode of
conidial development, the genera are placed under eleven sections.
This classification was proposed by Hughes in 1953, which was
strengthened by Tubaki (1958) and Subramanian (1962-1965). In the
older classification of Saccardo, the Hyphomycetes were identified
by the shape? of the mature conidia i.e., conidial morphology. We-
shall not enter into a detailed discussion of the two classifications^
Some important genera of Hyphomycetes are described below.
genus ALTERNARIA
perennate the fungus in absence of the crop. When the potato crop
is sown and the leaves are
formed, the conidia reach the
leaves through wind and germi¬
nate. The germ tubes enter the
leaves through stomata or by
direct penetration of the epider¬
mis, and form inter or intra¬
cellular mycelium^ The myce¬
lium secretes enzymes and toxins
which kill the cells. The fungus
derives nutrition from these dead
cells. When the cells die, the
leaf spot symptoms appear. Cla-
vate conidia having both trans¬
verse and longitudinal septa
(Fig. 8.2) are formed on the F'g- 8.2. Conidia and conidiophore of
hyphae. The conidiophores show Alternaria solanL
‘knee’-like swellings which indicate the position of the detached
conidia. The conidia are wind-disseminated and in this way the
disease spreads to more plants throughout the season. In the absence
of the host plant, the hyphae or conidia remain in the fallen leaf
tissues or in the soil.
Control Measures
1. Field sanitation and rotation of crops can minimize the disease.
2. Regular spray of fungicides like Dithane Z-78, controls the dis¬
ease.
genus PIRICULARIA
Conidia are pyriform (Fig. 8.3) usually bi-septate and have a small
hilum at the base. These are borne apically on conidiophores which
emerge through the stomata. P. oryzae causes the ‘blast disease’ of rice.
BLAST DISEASE OF RICE
Pathogen: Piricularia oryzae
This is the enemy number one of rice crop and occurs in India
frequently.
Symptoms. Spindle-shaped spots (Fig. 8.3), ash-coloured in centre
with brown margins, appear* on the leaves, culms, glumes, and the
stalk (neck) of the panicle. The ears are also infected and bend down¬
ward due to rotting of the stalk.
128 ,
A Textbook of Fungi Bacteria and Viruses
geiius FUSARIUM
Species of Fusarium are important because they cause root rot and
wilt diseases of several plants. In wilt diseases the leaves droop down
as happens in unwatered plants. The water absorbed by the roots is
not transported to the leaves due to the blockage of the xylem vessels
by the hyphae. The leaves,due to shortage of water, droop, dry and
die. All the wilt-causing species of Fusarium are called as F. oxy-
sporum. The hyphae are septate and branched. The conidiophores
are short and consist of a group of spore-producing cells called pbia»
tides (Fig. 8.5). Each of these form curved, sickle-shaped macroco-
midia and globular micro-conidia. The spores are held together in
• Slime
Microcoriidium
Macroconidium
Conidiophore
Hypha
Agar surface
Fig. 8.5. Fusarium sp. The macro-and micro conidia borne on conidiophores
in a slimy mass.
slime and, therefore, these spores are called slime spores. These are
not wind-disseminated. In Alternaria, Piricularia and Helminthospo-
rium, the spores are dry spores and are easily blown by wind.
class—COELOMY CETES
Majority of the genera are parasitic and cause leaf spots. Pestalo-
tiopsis, Colletotrichum and Gloeosporium are the important genera.
Control
1. The blighted leaves should be collected and burnt to prevent in¬
fection next year.
2. Spraying of the crop should be done frequently by Bordeaux
mixture.
3. Water-logging favours the disease and, therefore, should be
avoided.
132 A Textbook of Fungi, Bacteria and Viruses
genus GLOEOSPORIUM
The conidia are hyaline, elongated, with rounded ends and slightly
narrower in the middle. G. musarum is the most important species
which causes ripe rot of banana fruits. Brown spots appear on the
skin of the fruits which become sunken and the rotting encroaches
the edible portion. Acervuli appear on the sunken black areas.
genus COLLETOTRICHUM
The pycnidia vary in size* shape, colour and consistency of the wall
and provide the basis for identification of the genera. The conidia,
formed inside pycnidia, may be hyaline or coloured, septate or
unseptate. Phorna, Phyllosticta, Ascochyta, Diplodia, Septoria etc.
are the important genera.
Phorna, Phyllosticta and Macrophomina, all the three genera have
similar small, dark, globose, ostiolate pycnidia, sunk in the substratum
(Fig. 8.9). Cooidiophores are short and bear hyaline spherical or
oval, one-celled conidia. There is no real distinction among these
genera. If the fungus grows on leaves it is called Phyllosticta; on
stem, it is Phorna or Macrophomina,depending on the conidiai size. It
is now felt that the three genera should be called by any one name.
If the leaf spots bearing pycnidia are teased by a needle and examin¬
ed under the microscope, a stream of minute conidia is seen lowing
out of the pycnidia.
9. BETEROTHALLISM, HETEROKARYOSIS
AND PARASEXUAL CYCLE
HETEROTHALLISM
Bipolar Heterothallism
Fungi in this category have two mating types, each containing
genetically different nuclei. The sexual compatibility is controlled by
a pair of genetic factors A and a located at the same locus on diffe¬
rent chromosomes. This is therefore also called as 4Two allele hetero¬
thallism.” During meiosis, the two chromosomes, containing the
alleles A and a, are separated in the haploid spores (germ spores,
ascospores or basidiospores). The spores give rise to two types of
thalli, which must come together to bring together the two nuclei
carrying the compatibility factors A and a. The two mating types are
designated as (+) and ( —) strains.
The 4Two allele” or “bipolar” heterothallism is found in Muco-
rales (Mucor, Rhizopus, Phycomyces), Ascomycotina (Neurospora
Ascobolous), Basidiomycotina (.Puccinia graminis and the smut fungus
Ustilago levis).
Tetrapolar Heterothallism
Fungi in this group form thalli of four mating types. This type of
heterothallism is governed by two pairs of compatibility factors Aa
and jBb, located at different chromosomes, which segregate indepen¬
dently during meiosis. If crossing over occurs between the mating
type loci, four types of segregations (AB, Ab, aB, ab) are possible
depending on the chromosomal arrangement.
A a
\
B AB aB
b Ab ab
Thus four types of spores (,AB, Ab, aB and ab) are formed which give
rise to four types of thalli. Only those thalli that have nuclei carrying,
opposite genes for both the factors, can mate. The resulting zygote
must have the genotype Aa> Bb.
Majority (63%) of the heterothallic Basidiomycotina are tetrapolar
forming four types of basidiospores.
Heterothallism, Heterokaryosis and Parasexual Cycle 137
However, if crossing over does not take place, only two types of
spores (AB and ab or Ab and aB) are formed and only two types of
thalli are produced. Since it is governed by two factors it is called
tetrapolar.
Secondary homothallism. In some bipolar species the two
nuclei, which should give rise to hyphae of two mating types, are
contained in the same spore. Thus,the hypha produced behaves as
homothallic though involves genetically different nuclei. This situa¬
tion is termed “secondary homothallism.”
Korf and Hartman recommended that the terms Homothallism
and Heterothallism should be abandoned. Esser (1959) suggested the
use of the terms monoecious and dioecious, as done in higher plants.
Call by whatever Term’ you please, the function of heterothallism
remains unaltered. Heterothallism is a device for achieving outbreed¬
ing, which is a genetic desirability. Homothallism brings in inbreed¬
ing and provides no chance for genetic change.
HETEROKARYOSIS
PARASEXUALITY
Until 1944, the sexual cycle was the only means of exchange of
genetic material. It is to the credit of microbial geneticists that a
series of novel methods of genetic recombination are now known in
bacteria, which do not involve karyogamy and meiosis. These are
transformation, conjugation, transduction, lysogeny and sexduction,
which differ from the standard sexual cycle. The mechanism of each
of these ‘alternatives to sex’ is described in detail later in the
chapter on reproduction in bacteria (chapter 15).
A similar alternative to sexual reproduction was discovered in
the imperfect fungus Aspergillus nidulans in 1952 by Pontecorvo and
Roper of Glasgow. This they called parasexual cycle. In this,
genetic recombination occurs in vegetative cells by the mechanism of
Homozygous
diploid nucleus
Haploid nuclei
4
Sorting out of Hapiodization and formation of
diploid strains haploid strains through haploid conidia
through diploid
conidia
Haplodization
The diploid colonies show appearance of sectors on the Petri plate*
which produce haploid conidia. This indicates that some diploid
nuclei must have undergone haplodization forming haploid nuclei,
which later get sorted out in haploid conidia. Some of these haploids
are genetically different from the original haploid parental nuclei.
This is because of the recombination that occurred during the mitotic
crossing over.
Haplodization occurs at a constant frequency of 1G“3 per nuclear
division. The haplodization occurs not by a reduction division
(meiosis), but by aneuploidy-—a phenomenon in which chromosomes
are lost during mitotic divisions. It happens like this. During mitosis
of the diploid nucleus, the chromatids fail to separate (non-disjunc¬
tion) in the anaphase stage. One daughter nucleus gets one chromo¬
some more (2n4-l), while the other gets one chromosome less
(2/i—I) than the normal 2 sets of chromosomes (In). Both the
daughter nuclei are called aneuploid. The deficient aneuploid
nucleus (2/2 — 1) may lose more chromosomes in the successive
mitotic divisions and finally reduced to haploid state (jn). Mitotic cros¬
sing over and haplodization occur also with the diploid hymozygous
nuclei but, since, the two nuclei are similar, crossing-over products
or the haploid nuclei formed by haplodization, are genetically no
different from the haploid parent nuclei.
The parasexua! cycle, thus, like the sexual cycle, involves plasmo-
gamy, karyogamy and haplodization but not at a specified time or
place. Every step differs drastically. The differences between the two
cycles are given in the Table 9.1.
Table 9.1. A comparison between the sexual and the parasexual cycle
Zygote usually persists for one gene- Zygote persists for many generations
ration only. by mitotic divisions.
over during meiosis. The crossing over which is a rare event and occurs
over occurs in all chromosome pairs in a single chromosome arm. Haplo-
and is accompanied by a reduction dization, unlike meiosis, is indepen¬
in the chromosome number. dent of crossing over.
They can colonize rocks, but are also found growing on fertile soils.
The tree trunks on hills are the most common sites of lichen growth.
In some places the growth is so luxuriant that lichens are used as
fodder for cattle. But, lichens are most sensitive to air pollution and
probably that is the reason they are not found near cities (Nash III
1976).
The Thallus. The numerous types of thalli that the lichens exhi¬
bit, can be grouped under three morphological categories. These are
Foliose (leaf-like), Crustose (crust-like) or Frnticose (branched—
Upper
cortex
Apothecia
gal layer
Medullary
layer
Foliose
Lower
cortex
Rhizines
Apothecia Upper
cortex
Algal
Crustose layer
Medullary
f>rin H| layer
Algal
cells
Hyphae
Fruticose
Fig. 10.2. The three morphological forms of Lichens and their anatomy.
Reproduction
Asexual reproduction. Asexual reproductive methods (including
vegetative reproduction) are the main source of multiplication (Fig.
10.3). The sexual reproduction is insignificant in the life of lichens.
Fragmentation. Death and decay of old parts of the thallus pro¬
duce smaller pieces which give rise to new thalli.
Soredia (sing, soredium). These are minute outgrowths—formed
on the upper surface of the thalli in distinct pustule-like sori called
sorelia (sing, sorelium). The large number of soredia form a
powdery coating on the surface. Each soredium consists of a few algal
cells surrounded by a mass of hyphae. These are dry and, therefore,
easily blown by wind. Each soredium gives rise to a lichen thallus.
Isidia (sing, isidium). These are stalked outgrowths of the thallus
containing masses of algae and fungal hyphae surrounded by a portion
of the thallus cortex. These are not detachable. Accidental breakage
The Lichens 145
tsidia
Pycnidium fragmentation
The algal and fungal components of several lichen genera have been
isolated and grown in culture. It has been found that they grow extre¬
mely slowly—not more than 1-2 mm in a year. The mycobionts are
dependent for growth on external supply of vitamins. e.g. thiamine
and biotin. However, when,both the components were grown together,
the algae died. Sometimes, the dead algal cells were found to be filled
with fungal hyphae. Death of algal cells was noted when the myco-
biont of the lichen Collema tenax was grown with its algal compo¬
nent Nostoc or with any other alga isolated from other lichen genera.
The Lichens 147
It was explained that the first reaction between the two partners is
that of parasitism of the alga by the fungus. The algae which survive
this parasitism enter into lichen association.
Ahmadjian, however, in 1962 succeeded in synthesizing the lichen
Acarospora fuscata by growing its fungal and algal components to¬
gether on a medium on which each alone failed to grow. This gave a
valuable indication that the condition which was unfavourable for
each of them independently was favourable for a joint growth and
formation of lichen.
Thus, a very delicate balance exists between their association and
if there is any disturbance, dissociation results. Their association is a
“marriage of convenience.” Under the luxuriant conditions in labo¬
ratory culture, the lichens dissociate and this creates a big hurdle in
the study of their biology.
11-THE BACTERIA
The cell wall of bacteria (and also of blue-green algae) differs from
rest of the plants in being made up of mucopeptide and not cellulose.
This difference provides a site where bacterial pathogens can be
attacked by antibiotics without damaging the diseased eukaryotic
plant or animal.
Mucopeptide is a polymer made up of alternating units of NAG
(N-acetyl glucosamine) and NAM (N-acetyl muramic acid) joined
by 3, 1-4 linkages. NAG and NAM are amino sugars (see struc¬
ture in Fig. 11.6). The mucopeptide chains are laterally linked by
short chains of amino acids, which originate at the carboxyl group
of the muramic acid molecules. The amino acid chains are linked
154 ,
A Textbook of Fungi Bacteria and Viruses
CH3CHC00H, CH3CHC00H
f L-Ala
1 1
-NAM-NAG-NAM-NAG-
acid. Some of the amino acids are D-isomers, which is quite pecu¬
liar, as in normal metabolism only L-amino acids are used.
The cell walls of Gram +ve and Garm —ve bacteria differ in
their chemical composition (Fig. 11.7). The wall of the Gram +ve
bacteria is homo¬
geneous containing
85% or more of
- - - - Cytoplasm * mucopeptide and
Gram + ve Gram—ve simple polysacch¬
aride, like teichoic
Fig. 11.7. The cell wall of Gram +ve and Gram —ve acids (teichos =
bacteria. M. mucopeptide, LPr lipoprotein, LPs, wall) which are
Lipopolysaccharide. polymers of ribitol
and glycerol phosphates. Teichoic acids serve as antigens and also
regulate entry of ions. The cell wall of Gram —ve bacteria contain
only 3-12 % mucopeptide, the rest being lipo-protein and lipo-poly-
saccharides. The wall of Gram — ve bacteria, in electron microphoto¬
graphs appears tripartite i.e. 3-layered. The cell wall differences of
The Bacteria 155
Smear (heat-fixed)
The Nucleoid
In bacteria, the genetic material (DNA) is not bound with proteins
to form chromosomes. The genome consists of a single closed ring
1000/x long. Thus in a bacterial cell the DNA molecule is over a
The Bacteria 157
thousand times longer than the cell itself. The DNA helix must un¬
twist itself every half an hour at every replication during cell division.
The expression “the tangled skein of life” for the DNA, could have
not been more appropriately coined. DNA molecule is attached to
the cell membrane possibly at the mesosome. The DNA, in fulgen-
stained preparations, is seen concentrated forming a gel-like structure
less dense than the cytoplasm. This concentrated structure is called
nucleoid or nucleoplasm. There is no nuclear membrane, and spindles
are not formed during cell division.
Ribosomes
These are small particles (100 A in diameter), which, along with
the reserve materials, give a granular appearance to the otherwise
158 ,
A Textbook of Fungi Bacteria and Viruses
Reserve Materials
Granules. The cytoplasm itself is a homogeneous aqueous solu¬
tion of soluble proteins, enzymes, cell solutes, inorganic ions and
metabolites of small molecular weights. Under electron microscope
the cytoplasm appears granular due to the reserve materials. Some
reserve materials lie in a state of fine dispersion in the cytoplasm.
The reserve materials can be classified into 3 categories:
1. Organic polymers—polys^iCcharidQS, lipids etc.
2. Inorganic metaphosphate granules—Volutin granules which are
polymers of phosphate inso¬
luble reserves.
3. Elemental sulfur—present in sulfur-oxidizing bacteria; serves
as energy reserve.
Nature of metabolism
Mechanism of Nature of cell
Bacterial types Non-photosyn¬
movement wall
Photo synthetic
thetic
EUBACTERIA
(b) Bacillus (pi. bacilli). These are small rods which may be flagel¬
lated or non-flagellated. The flagella may be present at one end, on
Spirillum Vibrio
Staiked-bacterium
Rhodomicrobium
Actinomycete Actinomyces
both ends or all over the body. This number may also vary from one
to many. The flagellation (Fig 12.2) is described as monotriehous
(single flagellum on one side), lophotrichous (a tuft of flagella on
one end), amphitrichous (tuft on both the ends) or peritrichou*
(flagella all over the cell).
(c) Vibrio (pi. Vibrios). These are short, curved, comma-shaped,
flagellated and monotrichous i.e. one flagellum present at one end.
162 A Textbook of Fungi, Bacteria and Viruses
(d) Spirillum (pi. Spirilla). These are spirally twisted and non-
flexible, may be flagellated or non-flagellated.
Peritrichous
Fig. 12.2. Flagellation in bacteria.
(e) Stalked bacteria (order Caulobacteriai^s). These are single-
celled, f *;alked-bacteria (Fig.
12.3). A tuft of flagella is pre¬
sent on one side (lophotrich-
ous). The staik is a part of the
cell as m Caulobacter but it is
a secreted product in the genus
Gallionella. The base of the
stalk is knob-like and sticky.
Thus several stalked cells are
joined by their knobs to form
a^rosette like structure (Fig.
Fig. 12.3. Caulobacter, a stalked bacterium. 12.3). Cell division occurs by
A. rosette formed by adhesion of the bases transverse fission which is
of the stalks, B. a single cell showing divi¬
not binary fission because the
sion by transverse fission.
Morphological Types of Bacteria 163
basal and apical daughter cells are different in structure. The apical
cell, which is equipped with polar flagella, swims away and later
develops a stalk.
(/) Budding Bacteria (order Hyphomicrobiales). Example—
Rhodomicrobium. The cells are like a foot-ball having a swollen part
and a thinner tube. The tube gradually increases in length and swells
to form a new globular cell. Ultimately a network of cells is formed.
(order MYXOBACTERIALES)
These bacteria lack a rigid cell wall and also the flagella. They show
gliding movement. The individual cells are cigar-shaped, which
divide by binary fission. The cells live as a colony in a common slimy
mass. At the time of fruiting, the cells aggregate and heap up to
164 A Textbook of Fungi, Bacteria and Viruses
form characteristic fruit bodies (Fig. 12.4). The slimy mass hardens
to form tree-like branches
Germinating which bear bri§ht|y
CySt coloured oval or spherical
/ cysts. Each cyst consists of
,anSr several hundreds of cells.
On germination the cyst
wall breaks and the cells
glide out. The myxobac-
teria live mostly in soil,
dung and water. The
Fig. 12.4. Chondromyces, a myxobacterium. aquatic forms are impor-
A. fruiting body consisting of cysts borne tant cellulose-decomposers,
on hardened slime, B. germination of cysts gome cause most destruc-
andjreiease of cigar-shaped cells of Chond- . diseases of fishes
romyces.
These are relatively large, helical or spiral bacteria which lack a rigid
cell wall (Fig. 12.5). Flagella are absent. The locomotion is not by
spirally around the cell and anchored in the cytoplasm at each end.
The contraction of the axial filament is caused by its contractile pro¬
teins. The spirochaetes divide by binary fission and do not produce
any resting spores.
The important genera are Spirochaeta and Cristispira. They live in
muddy waters and cause diseases like jaundice and syphilis in man.
NAG
(Gly)s
-NAG-
L-FORMS
-diffuses out
A B
Table 13.1
Sphaeroplast Protoplast
3. Properties associated with 3. Properties associated with cell wall are corn-
wall (viz., cell rigidity, pletely lost,
osmotic stability, antige¬
nic property, resynthesis
of flagelia, flagellar move*
ment) are lost but not
completely.
Sphaeroplast Protoplast
5. Can resynthesize the cell 5. Fail to resynthesize the wall because the
wall wheii the sphaero- mesosome, which is required for synthesis
plasting agent is with¬ of cell wall material, is extruded out.
drawn. This is because
the mesosdme, Whien was
retained, is available for
cell wall synthesis.
Characteristics
1. Mycoplasma lack cell wall (Fig. 13.5), have no shape (pleomor¬
phic)—hence called “jokers in the microbiological pack.” Do not
revert to normal cells and are osmotically stable.
Bacteria Without Walls *
Plasma membrane
Genome (DNA)
Ribosomes
Interrelationships of Mycoplasma
The genus Mycoplasma was included among bacteria under a sepa¬
rate order Mycoplasmatales. However, in 1966, the International
Mycoplasma
4
Antibiotic Antibiotic treatment
treatment stopped
*
Bacterial cell^
N
(Gram-t-ve or
Gram - ve)
ECONOMIC IMPORTANCE
Plant Diseases
Witches broom of potato and opuntia, Aster yellows, Com stunt
and the Sandal-spike disease (threatening the Sandalwood industry
in our country), are caused by Mycoplasma.
Human Diseases
The human infertility is caused by 3 species of Mycoplasma, vzz.,
M. hominis, M. fermentans and T. mycoplasma. Gnarpe and
Friberg (1973), two Swedish doctors isolated T. mycoplasma: from the
sperms (the head and middle region) collected from infertile (barren)
human males. Doxyccyline treatment cures infection by T. myco¬
plasma.
Animal Diseases
Bovine pleuropneumonia and infectious agalaclea are important
animal diseases.
14. BACTERIAL NUTRITION AND MINERAL
CYCLES
bacteria.
(0 In the first step ammonia is oxidized into nitrite by species of
the genus Nitrosomonas. The energy liberated is the sole source of
energy for growth of these bacteria. The carbon is obtained from
C02.
NH3+1|02 — - N02“ + H20 + H++energy
d0 In the second step, the nitrite is converted into nitrate. This is
brought about by species of the genus Nitrobacter, which use this
energy for growth. Here also carbon is derived from C02.
N02 + i02 ■—> N03" + energy.
(c) Sulfur Bacteria, d) Oxidation of elemental sulfur. ThiofeaciK
lus thio-oxidans obtains energy from oxidation of elemental sulfur.
Sulfuric acid is produced. This bacterium thus can survive in ex¬
treme acidic environment.
S°+H20 + 1£02 — -* H2S04 + energy.
d0 Oxidation of H2S to S—Beggiatoa uses the energy from oxida¬
tion of H2S for growth, and the product sulfur is stored as granules
in its filaments.
H2S + 0-* H20 + S° t energy
(d) Iron bacteria. This is the most interesting and most simple oxi¬
dation. Example—Ferrobacillus, The Ferric iron is deposited as7 in¬
soluble ferric hydroxide.
Fe+2-> Fe+s+e~
4. Chemosynthetic heterotroph (Chemoorganotroph). Ener-_
gy is obtained from oxidation-reduction reactions using organic com¬
pounds as the oxidizable substrate. Animals, fungi and most bacteria
(e.g„ Escherichia coli) are chemoorganotrophs.
The bacteria in this category secrete extra-cellular enzymes which
degrade complex nutrients,~ carbohydrates, proteins, fats etc. into
simple smaller units which are absorbed and oxidized for release of
energy. If Oa serves as the ultimate hydrogen acceptor the energy-
yielding oxidation is called respiration; if it is an inorganic substanc
other than oxygen, it is called anaerobic respiration. It is called
fermentation if an organic substance serves as the hydrogen accep¬
tor.
Bacterial Fermentations
Carbohydrates are the most important substrates of bacterial fer-
176 ,
A Textbook of Fungi Bacteria and Viruses
Bacterial Respiration
Aerobic respiration. The oxidation of organic compounds by
oxygen is called aerobic respiration; C02 4 HaO are the end products.
The role of bacteria (as scavangers) in disposal of organic debris is
of great significance. They are capable of oxidizing any organic com¬
pound found in the living world.
Anaerobic respiration. Species of the genus Desulfovibrio oxidi¬
se organic compounds anaerobically, using sulfate as the hydrogen
acceptor (oxidant). Another important anaerobic respiration involves
the use of nitrate as hydrogen acceptor. It is reduced to N2 or NH3
by the denitrifying bacteria. It is important to note that the sulfate
reducers can not use oxygen even when available, but the denitrifying
bacteria use nitrates only in the absence of oxygen. The denitrifying
bacteria restore the nitrogen gas to the atmosphere and, thus, are of
great importance in maintaining the nitrogen cycle.
The elements present in the living matter are returned to the atmos¬
phere from which they were taken. The part of the earth, which
BIOSPHERE
ASEXUAL REPRODUCTION
Fission. The bacterial cell divides into two by fission (Fig. 15.1)
Mostly it is binary fission in which the two daughter cells are identi¬
cal. Division is a very quick process and is completed in 30 minutes. Ir
6 hours a million bacteria are produced from a single parent cell.
Fig. 15.1. Binary fission. One bacterium gives rise to two identical cells.
ing fork. It is that point where the two strands become four. In-each
of the two daughter circular ‘chromosomes5, one strand is derived
from the parent and
^— -Initiation point the other is new. This
type of replication in
if \\ which one strand is
H JJ old and the other is
newly synthesized is
, called the semicon-
6
1 servative replica-
tion. This mechanism
°f replication was
(f/Y )) proposed by Cairns
JJ m 1963 and is known
o .• r , as Cairn’s model.
>-^ —- Replication fork n n .
| Cell division. A
y peripheral ring of
plasmamcmbrane in-
vaginates and grows
/A\ \\ centripetally to form
11 1) a double membrane
Septum (Fig. 15.3).
Wall material is depo-
\ sited between the two
membranes of the
septum. The two
JJ Y\ events i.e. septum for-
[( jJ mation and the cross
vv JJ wall synthesis occur
\ simultaneously. The
2 DNA molecules cell division is follow¬
ed by separation of
Fig. 15.2. Showing replication (duplication) of the daughter cdls as
the bacterial chromosome. The Cairn’s model. unicellular bacteria
by dissolution of the
matrix between the newly synthesized walls. However, in the Actino-
mycetes, cell division occurs without separation and thus the cells are
not separated.
Higgins and Shockman (1971) have proved that mesosomes are
involved ir. the initiation of both the events viz., the DNA replication
Reproduction in Bacteria 181
Fig. 15.3. Septum and cell wall formation during bacterial cell division.
Coccus
Streptococcus
Fig. 15.4, Binary fission in Cocci. Successive divisions occur in more than
one plane and give rise to various forms of Cocci.
182 A Textbook of Fungi, Bacteria and Viruses
and the septum and wall formation. These two events occur simulta¬
neously during cell division.
In the spherical forms (cocci) divisions may occur in any plane but
in bacilli, divisions occur perpendicular to the long axis of the cell.
In cocci divisions occur as follows (Fig. 15.4).
enclosed cell moves out. Cysts are formed in Azotobacter and Jvlyxo
bacteriales e.g. Myxococcus, Chon-
dromyces etc.
Fragmentation, Sporangiospo¬
res and Conidia. These methods
of asexual reproduction are common
in Actinomycetes and are same as Fig. 15.6. Cyst formation and
seen in the fungi (Fig. 15.7). In fact germination.
before their prokaryote cell-stru¬
cture was discovered, Actinomycetes were placed in fungi.
Arthrospores
Actinomyces
SEXUAL REPRODUCTION
(ii) the extremely rapid growth and small size. Millions of bacteria
are easily handled and held in a drop of fluid.
(7/7) the variety of genetic mechanisms that operate in bacteria
greatly facilitated genetic analysis, and
(?v) it proved easier to relate the genetic phenomena to the
structure and function of the biologically important macro¬
molecules.
The most important achievement was the discovery that the sexual
reproduction (karyogamy followed by meiosis) was not the only
method of genetic recombination and that nature employs several novel
methods, hitherto undiscovered, to achieve the same end. There
exist four mechanisms of genetic transfer in bacteria, which differ in
the mode in which DNA is acquired by the recipient bacterium.
These are:
Transformation. In this, the DNA is absorbed from the external
medium. This is the method employed in laboratory to bring about
recombination; but it may be occurring, in nature as well.
Conjugation. The recipient female cell (F~) receives the DNA
from a donor bacterium—Hfr, a male with high fertility rate, through
cellular contact. Though it appears similar to conjugation of higher
forms, bacterial conjugation is peculiar and different.
Transduction. The DNA is acquired through a virus. Thus, it is
a “phage-mediated genetic transfer.”
Lysogeny. It involves association of genetic material of a virus
with that of the bacterium. Although different from above methods,
it also provides a permanent genetic modification of the bacterial
genome.
We shall adopt a historical approach in the study of sexual repro-
ductionr as it will give a better perspective and make the subject more
clear.
Genetic Recombination. When the DNA enters the recipient
cell by any of the three mechanisms (through medium, by conjuga¬
tion or through a bacteriophage), it can synapse with the homologous
region of the recipient genome and undergo recombination to give
new genome types. The reciprocal product of the recombination
process is broken down by DNAase enzyme.
TRANSFORMATION
Historical
(i) Griffith observed Transformation in bacteria. In 1928,
Reproduction in Bacteria 185
.•.-I__
Healthy
Dead
Virulent
Healjthy
Heat-killed virulent
Virulent strain
(capsulated)
n
Virulent heat-killed Disrupted cells
bacteria (capsulated)
nnrsrrrTT
Purified DNA
Mechanism of Transformation
A fragment of DNA, present in the medium, gets adsorbed on the
recipient cells and then enters the bacterial cells (Fig. 15.10). Inside
the cell, it replaces the homologous part of the bacterial genome.
This results in the development of new characters which pass on to
the progeny cells as stable and heritable character.
Virulent progeny.
CONJUGATION
Historical
Discovery of mutation in bacteria by Delbruck and Luria in
1943. An important aspect of study in genetics is mutation, which
is sudden spontaneous heritable change of the genetic material.
Edward Tatum and Joshua Lederberg
ally, both the strains were unable to grow on minimal media* (which
O
lack any growth factor). But when the two strains were mixed and
A-B-C+D+xA+B+C-D-j
| (SRPs) (ssn
When tested for streptomycin and phage resistance, (see lower part
of Fig. 15.11) these markers also appeared in hew combinations.
None of the parents were double sensitive or double resistant (SSPS
or SfPr) but, as seen in Fig. 15.11, the two colonies (3 and 4)
were double resistant SrPr as they grew on both streptomycin and
phage-containing media. Three colonies viz., 6, 7 and 8 were double
sensitive as they did not grow on any plate.
Mechanism of Conjugation
The exact mechanism of the genetic exchange was worked out by
three groups of scientists led by Lederberg in Wisconsin, USA,
Hayes in London^and Woolman in Paris. That transformation was
not involved, was proved by the finding that cell-free extracts of one
F+(male) X F-(female)
F+(male) F+(male)
genome
Fig. 15.13. Conjugation between high fertility male (Hfr) and female;
(F“) cells, resulting in production of recombinant female.
192 A Textbook of Fungi, Bacteria and Viruses
^---
Separation
Faulty separation
(Sexduction)
TRANSDUCTION
Historical
Lederberg and his student Zinder in 1951 started looking for re¬
combination in Salmonella typhimurium—the mouse typhoid bacte¬
rium. They used the same techniques which Lederberg and Tatum
had used with E. coli. They obtained nutritionally-deficient mutants
(auxotrophs) which failed to grow on the minimal medium. When a
mixture of the two mutants were plated together, recombinants
appeared in a few cases but not with other strains. When they analy¬
sed the cause they discovered a new type of gene exchange, which
involved the mediation of bacteriophages. Zinder and. Lederberg
described this new method of gene acquisition in 1952 as Transduc¬
tion.
In their experiments, conjugation was ruled out by the following
experiment (Fig. 15.16). A U tube was taken which had a sintered
glass filter between its two arms, through which bacteria could not
pass. Two auxotrophs (^nutritionally deficient mutants) were grown
in the two arms (strain A in one arm and strain B in other). The
medium from one could freely go into the other arm. Recombinants
appeared in the bacterial population of one of the arms only. Why?
Transformation was also ruled out as DNAase (the DNA splitting
enzyme) had no effect on the gene transfer.
ABORTIVE TRANSDUCTION
Sometimes the DNA, brought by the phage, does not integrate with
the genome, of the recipient bacterium and expresses itself indepen¬
dently. It does not replicate and so it passes on during binary fission
to only one of the two daughter cells. For example the motile
character transduced in a non-motile strain, at the time of cell division
is passed on only to half of the progeny. The transduced DNA goes
to only one of the daughter cells, as it is neither integrated with the
chromosome nor it replicates itself.
LYSOGENIC CONVERSIONS
Bacteria as Pathogens
First bacterial disease, the 'fire blight’ of pear, was described in
1878 by T.J. Burrill, —only two years after Koch’s demonstration of
the role of Anthrax bacillus in animal diseases. F.F. Smith by 1900
established the role of bacteria as plant pathogens.
Species in all major families of higher plants are known to be
attacked by one or more bacterial pathogens.
Symptoms. The symptoms produced by bacterial pathogens are
not markedly different from those incited by fungi. The main symp¬
toms are:
Wilting—Pseudomonas solanacearum is most important which
causes wilt of several crop plants.
Blight—-Leaf blights are usually caused by species of Xanthomonas
and Pseudomonas.
X. orytae causes 'leaf-blight’ of rice.
P. tabaci—‘wild-fire’ disease of tobacco.
Leaf spot—Angular leaf spot of cotton Xanthomonas malvacearum.
Canker—Xanthomonas citri causes citrus canker.
Gall—‘Crown gall’, on over 142 angiospermic genera, is caused by
Agrobacterium tumefaciens. Bacteria are found in the peripheral zone
of the galls. Since growth patterns resemble those of animal and
human cancerous tissue, bacterial galls are most extensively studied.
Scab—Potato scab caused by Streptomyces scabies.
Although most bacterial pathogens induce one major symptom,
some diseases show more symptoms. In the bacterial blight of paddy
(X. oryzae), the disease syndrome comprises blight and wilt symp¬
toms.
Bacterial lesions are characterized by an overlying waxy layer. In
wet weather bacterial masses ooze out from the lesions and on
drying, get deposited as waxy scales. The bacteria cannot be dislod¬
ged unless softened by dew or rain.
teristic
pigments
6. Xanthomonas Wilt, leaf-spots, 47 •* Yellow Motile
blights
CITRUS CANKER
Pathogen—Xanthomonas citri
It is a Gram — ve bacterium which has two polar flagella. The
name Xanthomonas is derived from Greek word, Xanthus, which
means yellow, as the bacterium produces a yellow pigment in the
growth medium.
Symptoms
Lesions consisting of depressions with raised margins are called
canker. Cankers appear on all parts of citrus plants (Fig. 16.1). At
first, raised pustules are formed, which rupture and depressions
appear at these places. Yellow hallows surround the cankers on the
leaves but not on fruits. In cases of severe infections, the leaves fall
off causing defoliation of the plant Infected twigs dry up.
Disease Cycle
The bacterial cells can survive in soil for 4-5 years in the absence
of the host. Thus soil serves as the main source of infection. The
infected twigs, leaves and fruits carry the bacteria and cause infec¬
tions. The bacteria enter the leaves and other parts of the plant
through stomata or cut surfaces in a film of water. Wind is unimpor¬
tant in the spread of plant pathogenic bacteria. The bacterial cells
grow and multiply in the intercellular spaces and cause the symp¬
toms.
Control
1. Diseased plants should be removed and burnt as soon as detec¬
ted.
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Fig- 16.1. ''Citrus Canker" symptoms on leaves (A), and stems (B), caused by
Xanthomonas citri.
Pathogen—Pseudomonas solanacearum
It is a Gram —ve bacterium having two polar flagella. In addition
to tomato, the pathogen causes wilt of other plants also like potato,
egg plant (bringal), ground nut and banana. On the basis of their
host range, P. solanacearum is divided into three races 1, 2 and 3 bv
Buddenhagen and Kelman (1964). Race 1 has a wide host range while
race 2 and 3 infect only a few plants. The pathogen produces slime
which causes the wilt symptoms. Mutant strains, which don’t produce
slirne, cease to be pathogenic.
Symptoms. The most obvious symptom is the epinasty /.<?.,
drooping down of the leaves, petioles and branches, much in the
same way as seen when plants are not watered (Fig. 16.2). The other
important, symptom is the browning of the vascular tissues. Vein¬
clearing, chlorosis of leaves, defoliation and stunting of growth also
accompany wilting and browning symptoms.
Disease Cycle
The pathogen is common in soil everywhere. The bacterial cells
enter the roots through injured surfaces in a film of water, and reach
the xylem vessels, where they grow and multiply. Occasionally, they
form bacterial pockets in the root cortex. The slime, produced by the
bacteria in the xylem vessels, increases the viscocity of the sap. This
results in slow upward movement of water. The leaves get reduced
supply of water and the leaves show symptoms of drought. The
plants dry and die. The bacterial cells reach the soil after death and
decay of the host plant tissues.
Control
Use of resistant varieties, which are available, is the only control
measure.
] 7- VIRUSES—DISCOVERY, NATURE AND
BIOLOGICAL STATUS
Great fleas have little fleas upon their backs to bite them, little
fleas have lesser fleas and so ad infinitum. SwifWDe Morgan
f. C. Bawden
Viruses—Discovery, Nature and Biological Status 203
Structure of Viruses
Before the advent of the electron microscope also the structure and
shape of viruses was studied with great accuracy by indirect chemical
methods. The viruses were found to be of two shapes—rods and
spheres. But the development of the electron microscope and the
techniques of negative staining and double shadowing, developed
in 1940’s, enabled to take pictures—(electron microphotographs) of
viruses. The rod-shaped viruses turned out in these photographs to
be helical. Spherical viruses appeared to be polyhedrons—mostly
icosahedrons (made of 20 faces). The protein coat was showed to be
not a single shell but composed of several units called capsomeres.
Thus, the electron microscope made many familiar things new and
new things familiar.
Contributions of molecular virology to the biological world are
many. Studies with bacteriophages established the basic pattern of
virus replication; confirmed the nucleic acid as the genetic material
and contributed fundamental knowledge about the gene structure.
Some of the important discoveries are listed in a chronological
sequence in Table 17.1.
Study of viruses, (virology) became an independent science. Be*
cause of their importance as disease agents and also* because of their
use as excellent tools in genetic analyses, virology came to occupy a
central place in molecular biology.
Viruses—Discovery, Nature and Biological Status 205
Year Event
1886 Adolf Meyer of Germany proved that the Tobacco Mosaic disease
was infections; but any pathogenic organism could not be isolated.
1892 Ivanovvski of Russia showed that the infectious agent after passing
through most reliable bacteria filters retained infectivity.
1896 Beijerinck a Dutch scientist said for the first time that bacteria were
not associated with this disease. The pathogen was not particulate
but a contagium vivum fluidum —contagious living fluid.
1915 Twort, an English bacteriologist, observed death of bacterial colonies.
1917 d’Herelle, a French-Canadian scientist, also noted conversion
of turbid bacterial colonies to ‘glassy’ masses which contained dead
bacteria. He observed that cultures upto million-time dilutions re¬
tained the infeciivity. He coined the term ‘bacteriophage’—the
“bacteria-eater.”
1933 Schlesinger, a Hungarian refugee working in London, reported pre¬
sence of only nucleic acid (DNA) and proteins in bacteriophage.
However, its significance was not realized. That was not the age of
nucleic acids.
3935 Stanley an American biochemist-isolated and crystallized TMV and
claimed that viruses were made exclusively of proteins. The crystal¬
lization was an inanimate character. He was awarded Nobel prize.
1936 Bawden and Pirie of England demonstrated nucleic acids and pro¬
teins in viruses. They established that viruses were nucleoproteins.
1938 Delbruck, a Berliner working at California, discovered mutation in
viruses and initiated work with bacteriophage of Escherichia coli (T
series).
1942-48 Luria, an American, and Delbruck elaborated on the process of
replication of virus genome.
1951 Safleraian and Morris, American scientists, discovered Cyano-
phages, the viruses that attack blue-green algae.
1952 Zinder and Lederberg discovered Transduction (virus-mediated DNA
exchange in bacteria).
1952 Hershey and Chase, two New York scientists, showed that the pro¬
tein part of virus was noninfectious; and that the nucleic acid was
responsible for infection and heredity. Thus they corroborated the
earlier findings of Avery, MacLeod and McCarty that DNA was the
genetic material.
1953 LwofF and his group at Pasteur Research Institute, Paris discovered
temperate phages; initiated elucidation of the lysogenic cycle in
bacteriophages and the lysogenic conversion, a new type of gene
exchange in bacteria.
1955 Takahashi (of Japan) and Frankel Conrat (of USA) reconstituted
TMV from its proteins and RNA.
18. THE VIRION
Definition of Virus
According to Luria and Darnell (1968) “Viruses are entities whose
genome is a nucleic acid, either DNA or RNA, which reproduce inside
living cells and use their synthetic machinery to direct the synthesis of
specialized particles—the virions, which contain the viral genome and
transfer it to other cells”
Fig. 18.1. Showing the various shapes of viruses and their relative sizes.
Envelope-
Spikes
Enveloped virus
Host cell membrane
Membrane protein of virus
Viral genome
coat. The protein is of viral origin and the lipid is derived from the
host. One characteristic feature of the envelope is that it is covered with
The Virion 211
projections called spikes, about 10 nun long; the size varies in diffc-
rent viruses. The enveloped viruses are formed in three steps (Fig.
18.3): (0 the formation of nueleocapsid, (ii) the insertion of viral pro¬
teins in the host plasmamemhrane, and (ill), budding—the virion nucleo-
capsid enveloped by the modified plasma-membrane is budded off.
Classification of Viruses
Earlier, the viruses, on the basis of their hosts, were classified into
plant viruses, animal viruses and bacterial viruses (bacteriophages).
But with the development of knowledge of the physicochemical
characteristics, a new classification was proposed by Andre Lwoff and
Tournier in 1966, which has been accepted by the Provisional
Committee on Nomenclature of Viruses (PCNV), especially constitu¬
ted for this purpose.
The classification is based on the following criteria of the viruses,
(1) Nature of nucleic the acid—DNA or RNA.
(2) Symmetry of the capsid—helical, cubical or both (binal).
(3) Presence or absence of envelope around the capsid.
(4) Diameter of helical forms.
(5) Number of the capsomeres in cubical forms.
212 A Textbook of Fungi, Bacteria and Viruses
PHYLUM VIRA
I -
sub-phylum-Deoxyvira sub-phylum-Ribovira
class—Deoxyhel ica class—Ribohelica
order—Chaetovirales order 1 Rhabdovirales
class—Deoxycubica ,, 2 Sagovirales
order—Haplovirales class—Ribocubica
class—Deoxybinala order 1 Gymnovirales
order—Urovirales „ 2 Tagovirales
19. BACTERIOPHAGES AND THE TOBACCO
MOSAIC VIRUS
BACTERIOPHAGES
Viruses that attack bacteria are called bacteriophages (or only pha¬
ges), a term coined by d’ Herelle in 1917, which means bacteria-eater.
Bacteriophage T* (Fig. 19.1) has been studied in great detail and
will be used as a typical example of bacterial viruses. It has a head
and tail-like structure.
The head contains the Protein coat
double- stranded DNA.
How this DNA is packed
inside the head is not
known. The head has the
shape of two halves of an
icosahedron connected by
a short, hexagonal prism.
The tail is helical. The
presence of two types of
symmetry in the protein
coat is called binal sym-
metry and is found ex¬
clusively in bacteriopha¬
ges and cyanophages. The
tail is made up of a cubi¬
cal hollow tube surroun¬
ded by a contractile
sheath. On one end it is F»g 19.1. Diagram of r4 bacteriophage,
joined to the head by a
‘colar’ while at the other end itlias a hexagonal plate which has
six short, small fibers (‘tail pins’) at every corner and 6 tail-fibers.
While the long, tail fibers are associated with adsorption of the
phage on the bacterial wall, the short tail fibers hold the phage fast
to the host during sheath contraction and DNA injection. During ad¬
sorption, a pore is dissolved in the bacterial wall, probably by a
phage enzyme and the DNA is injected into the bacterial cell. Phages
have two types of life cycles, lytic cycle (exemplified by T series of
phages infecting E. coll) and lysogenic cycle (example, X lambda
phages). Details about these life cycles will be seen in chapter 20 on
the “Growth Cycle of Viruses”.
X PHAGE
CYANOPHAGES
The virus particles are non-motile and reach the hosts passively
through cell sap of the previous host, water, insects etc. The events
which take place during virus growth cycle are:
1. Adsorption of the virion on the host cell surface.
2. Entry of the nucleic acid or the whole particle (in animal virus)
into the cell.
3. Transcription and translation.
4. Genome replication.
5. Assembly of new virions and
6. Release of virions from the cell.
The whole process is called the virus growth cycle. It is of very
short duration for bacteriophages (completed in 20 minutes) but some
animal viruses have longer cycles. This agrees well with the similar
short or long life of their hosts.
through them the proteins. Some proteins are used as enzymes for
the viral DNA synthesis. Proteins of the coat are also synthesized.
Bacteriophage (T2)
Phage DNA
Bacterial DNA
Maturation—genomes Protein synthesis degraded
wrapped in protein coal
Enzymes
The Tobacco mosaic virus (see Fig. 19.3) has no specialized organ
for adsorption or injection as seen in the bacteriophages. So, the
virus reaches inside the cell as a complete particle through cell sap
via abr.asions of the host surface.
The protein coat of the virus is dissolved by host enzymes and the
synthesis of mRNA, proteins and the genome as well as the matura¬
tion of the virus particles occur in the nuclei and not in the cytoplasm.
The lysis of cells does not occur and the virus particles move into
•other cells of the plant through plasmodesmata and also possibly
through the phloem. It is not known, for sure, whether the virus parti¬
cles are translocated inside the plant as complete particles or as naked
RNA. Ultimately, the release of the virus particles occurs, however,
not through cell lysis but through sap from injured tissues.
21. VIRUS DISEASES OF PLANTS
Non-persistant
or Stylet-borne
viruses in
epidermis
Stylet in epi<
Circulating viruses
multiply in the body
of the aphid and
reaches the salivary
Qtanda
Circulating (Persistant)
viruses in phloem
Stytet reaches
phloem
into the phloem (sieve tube) to acquire these viruses. There is a latent
period between the acquisition of these viruses and the time when the
aphid becomes infective. The circulative viruses multiply inside the
body of the aphids and pervade the whole body; ultimately reaching
the salivary glands. The aphids remain infective throughout their life.
00 Leaf-hoppers. Leaf-hoppers show greater biological rela¬
tionship with the viruses than the aphids. There is no mechanical
transmission by them. Viruses, transmitted by leaf-hoppers, are not.
transmitted by other vectors. This is called vector specificity j.e.
only a particular species or race of the leaf-hopper can carry the
virus. Hereditary transmission of viruses to the off-springs of leaf
hoppers has been detected upto several (31) generations. The leaf
hoppers are also affected adversely by association with the viruses. It
has been found that the vectors die early and show decreased power
of reproduction. The various vectors (nematodes, fungi and insects)
and the viruses that they transmit are given in Table 22.1.
Nematode:
Xiphinema index Grape ‘fan-leaf’ virus
Fungi:
Olpidium brassicae Tobacco mosaic virus (TMV)
Tobacco stunt virus (TSV)
Lettuce big-vein disease virus.
Synchytrium endobioticum Potato virus X (PVX)
Insects:
Myzuspersicae (aphid) Potato leaf-roll virus.
Nephotettix apicalis (leaf-hopper) Rice dwarf disease virus
Cicudulina sp. (leaf-hopper) Maize streak virus.
Thysanoptera (thrip) Wheat spotted-wilt virus.
Bemisia tabaci (white flies) Tomato leaf-curl virus infecting
tobacco.
TOBACCO MOSAIC
Disease Cycle. The virus particles reach the plants first from
plant debris or through tobacco products carried by farmers for
smoking. Inside the host cells, the virus particles multiply and produce
large number of TMV particles. The virus particles spread to other
parts of the'plant i.e<9 become systemic, through plasmodesmata or
phloem. The virus particles are disseminated to other plants through
contact. The diseased leaves, used in the manufacture of cigars etc. by
carrying the viruses, serve as the main source of infection. The viruses
also remain in the fields in the plant debris on the ground. The parti¬
cles can remain viable outside the host tissue for upto 25 years.
Control, (i) Cultivation of tobacco on soils free from the
disease.
(ii) Chances of contact transmission through smoking or imple¬
ments should be avoided.
POTATO LEAF-ROLL
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INDEX