Kendriya Vidyalaya Vallabh Vidyanagar

Investigatory Project:
Sporulation of Fungi

Name: Prayag S. Soni

Class: 11th
Roll No. : 1104
Guided By: Mrs. Suroj Kumari
 Index
S No. Topic Page
No.
1) Certificate 2
2) Acknowledgement 3
3) Introduction 4
4) Theory 6
5) Structure of Fungi 7
6) Inside the Fungus 10
7) Types of Fungi 11
8) Sporulation of Fungi 17
9) Sporulation Process in 20
Yeast
10) Uses of Fungi 25

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S No. Topic Page
No.
11) Harmful Effects of Fungi 27

12) Experiment 30
13) Aim 30
14) Materials Required 30
15) Procedure 30
16) Observation 33
17) Conclusion 37
18) Precautions 38
19) Conclusion of Project 39
20) Bibliography 40

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 Certificate
This is to certify that Prayag Saurabh
Soni, a student of class 11th has
successfully completed the research on
the Investigatory Project on the topic:
Sporulation of Fungi under the guidance
of Mrs. Suroj Kumari during the year
2022-23 in partial fulfillment of Biology
Practical.

Mrs. Suroj Kumari Mr. P. K. Sah

(Subject Teacher) (Principal)

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 Acknowledgement
I would like to express my special thanks of
gratitude to my teacher Mrs. Suroj Kumari
as well as our principal Mr. P. K. Sah who
gave me the golden opportunity to do this
wonderful project on the topic: Sporulation
of Fungi which also helped me in doing a lot
of research and I came to know about so
many new things.
I am really thankful to them.
Secondly, I would also like to thank my
parents who helped me a lot in finishing this
project within the limited time.
I am making this project not only for marks
but to also increase my knowledge.
Thanks Again to all who helped me.

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 Introduction
Fungus, plural fungi, any of about 1,44,000
species of organisms of the kingdom Fungi,
which includes yeast, rusts, smuts, mildews,
moulds, and mushrooms. There are also
many fungus-like organisms, including slime
moulds and oomycetes (water moulds) that
do not belong to kingdom Fungi but are also
called fungi. Many of these fungus-like
organisms are included in the kingdom
Chromista. Fungi are among the most widely
distributed organisms on Earth and are of
great environmental and medical
importance. Many fungi are free living in
soil or water; others form parasitic or
symbiotic relationship with animals or
plants.

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This experiment has been conducted on the
fungi which we are surrounded by and can
see in everyday life (namely bread moulds).

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 Theory
Fungi:
Kingdom of saprophytic and parasitic spore
producing eukaryotic, typically, filamentous
organisms formerly classified as plants that
lack chlorophyll and include moulds, rusts,
mildews, smuts, mushrooms and yeasts.

Structure of Fungi:
The main body of most fungi is made up of
fine, branching and usually colorless
threads called hyphae. Each fungus will

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have vast numbers of this hyphae, all
intertwining to make a tangled web called
the mycelium.
The mycelium is generally too fine to be seen
by the naked eye, except where the hyphae
are very closely packed together. The picture
below was obtained through a microscope.
The hyphae are magnified 100 times life
size.

Fungal mycelium is mostly hidden from

human view, not only because of its small
size, but also as a result of its location. The
tangled mycelial mass is usually hidden deep
within its food sources, such as rotting

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matter in the soil, leaf litter, rotting wood or
dead animals. The mycelium remains
undetected until it develops one or more
fruiting bodies, containing the reproductive
spores.
They are usually produced at the surface of
the food source, rather than hidden within
it, to allow the spores to be shed and
carried away by the wind or by water, or
animals. The fruiting bodies are usually the
only indication we have that a fungus is
present. Like icebergs, they represent a tiny
fraction of the whole fungus, with the most
of it being hidden from view.

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 Inside the Fungus:
● Fungal cell walls are rigid and contain
complex polysaccharides called chitin
(adds structural strength) and glucans.
● Ergosterol is the steroid molecule in the
cell membranes that replaces the
cholesterol found in the animal cell
membranes.
● Fungi can be unicellular, multicellular, or
dimorphic, which is when the fungi is
unicellular or multicellular depending on
environmental conditions.
● Fungi is the morphological vegetative
stage consisting of a tangle of slender,
thread-like hyphae, whereas the
reproductive stage is usually more
obvious.

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 ● Fungi like to be in a moist and slightly
acidic environment; they can grow with
or without light or oxygen.
● Fungi are sporophyte heterotrophs in
that they use dead or decomposing
organic matter as a source of carbon.
The morphology of the mycelium, mode of
spore formation, and fruiting bodies form
the basis of the division of the kingdom into
various classes.

Types of Fungi:
1) Phycomycetes:
Members of phycomycetes are found in
aquatic habitats and on decaying wood
in moist and damp places or as
obligate parasites on plants. The
mycelium is aseptate and coenocytic.

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Asexual reproduction takes place by
zoospores which are motile or by
aplanospores which are non-motile.
These spores are endogenously
produced in sporangium. A zygospore
is formed by fusion of 2 gametes. These
gametes are similar in morphology
(isogamy) or dissimilar (anisogamous
or oogamous).
Example: Rhizopus (bread moulds).

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2) Ascomycetes:
Commonly known as sac-fungi, these are
mostly multicellular (Penicillium) or
rarely unicellular (Saccharomyces). They
are saprophytic, decomposers, parasitic
or coprophylus (growing on dung).
Mycelium is branched and septate. The
asexual spores are conidia produced
exogenously on special mycelium called
conidiophores. Conidia on germination
produce mycelium. Sexual spores called
ascospores which are produced
endogenously in a sac like asci. These
asci are arranged in a different type of
fruiting bodies called ascocarps.
Example: Aspergillus, claviceps,
neurospora.

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3) Basidiomycetes:
Commonly known forms of
basidiomycetes are mushrooms, bracket
fungi or puff balls. They grow on soil, on
logs, and tree stumps and inliving plant
bodies as parasites. The mycelium is
branched and septate. The asexual
spores are generally not found but
vegetative reproduction by
fragmentation is common. The sex
organs are absent but plasmogamy is
brought about by fusion of two
vegetative or somatic cells of different
strains or genotypes. The resultant
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 structure is dikaryotic which ultimately
gives rise to basidium. Karyogamy or
meiosis takes place in the basidium
producing four basidiospores. The
basidiospores are exogenously produced
on the basidium. The basidia are
arranged in fruiting bodies called
basidiocarps.
Examples: Agaricus, Ustilago, Puccinia.

4) Deuteromycetes:
Commonly known as imperfect fungi
because only the asexual or vegetative
phases of these fungi are known. When
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the sexual forms of these fungi were
discovered they were moved into classes
they rightly belong to. It is also possible
that the asexual and vegetative stage
have been given one name (and placed
under deuteromycetes) and the sexual
stage another (and placed under another
class). Later when the linkages were
established, the fungi were correctly
identified and moved out of
deuteromycetes. One perfect (sexual)
stage of members of deuteromycetes was
discovered; they were often moved to
ascomycetes and basidiomycetes. The
deuteromycetes reproduce only by
asexual spores known as conidia. The
mycelium is septate and branches. Some
members are saprophytes or parasites
while a large number of them are
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 decomposers of litter and help in
mineral cycling. Some examples are
Alternaria, Colletotrichum and
Trichoderma.

Sporulation of Fungi :
Sporulation refers to the formation of
spores from vegetative cells during
unfavorable environmental conditions. As
such, it may be described as an adaptive
response that allows the organism to survive
given adverse conditions (radiation, extreme
heat or cold, lack of nutrition etc.).

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Fungi are the most studied spore-producing
organisms in the world. They produce a wide
variety of spores that significantly vary in
size, shape and other surface features that
suit their environment (for dispersal etc.).
Whereas the spores produced sexually
(through meiosis) remain dormant for
survival (e.g. ascospores), those that are
produced asexually (mitospores) are for
dispersal.
Produced through mitosis, asexual spores
released in high numbers are genetically
identical. This allows them to play an
important role in reproduction when they
land on the appropriate substrate in the
environment following dispersal.

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 Chlamydospore:
This is a type of fungal spore that develops
from the hyphal structures during
unfavorable conditions. Chlamydospores are
characterized by a thick, melanized wall that
protects the contents of the spore.
Different types of fungi may produce
different types of spores.
The following are some of the main groups
of fungi and the type of spores they
produce:
● Zygomycota: Members of zygomycota are
known as zygomycetes. They produce
both sexual (zygospores) and asexual
(sporangiospores) spores.
● Ascomycota: Ascomycetes also produce
both sexual (ascospores) and asexual
(conidia) spores.

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 ● Basidiomycota: Compared to the other
groups of fungi, basidiomycetes are
largely known to produce sexual spores
that are known as basidiospores.

Sporulation Process in Yeast

(Saccharomyces cerevisiae):

Saccharomyces cerevisiae is also known as

Brewer's Yeast and is used for brewing,
baking and wine making.
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As with bacteria (Gram positive bacteria),
fungi do not necessarily start forming
spores when environmental conditions
change. Rather, they attempt to adjust to
the new conditions through a number of
strategies.
When nitrogen sources diminish,
Saccharomyces cerevisiae may respond by
going into a stationary phase or modifying
their morphology. However, in the event that
unfavorable conditions persist, spore
formation becomes necessary. This involves
a process known as meiosis.
Early Phase:
During unfavorable conditions, the
organism responds by entering the early
phase (first phase of sporulation) of
sporulation. During this phase, cell division
processes shift from the mitotic cycle in G1
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(an interphase stage) and enter the S phase
in meiosis.
Some of the events that take place during
early phase include:
● DNA replication
● Homolog recombination and pairing
(prophase)
This phase is therefore largely characterized
by changes in the machinery of the cell cycle
as well as altered RNA processing.
Middle Phase:
The middle phase is characterized by
various cytological events. Here, the meiotic
process results in the production of haploid
nuclei (four) that form four new daughter
cells. This involves the modification of the
spindle pole bodies to contribute to the
formation of prospores (membrane
compartments).
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Some of the other events that take place
during this phase include:
● Post-Golgi secretory vesicles are
redirected to promote the regeneration
and expansion of the new compartments.
● The prospores grow and engulf the
nuclei.
● Other organelles move to the
cytoplasmic space that lies between the
nuclear membrane and the prospores.
● The nucleus is completely separated
from the cytoplasm of the mother cell.
Late Phase:
This is the last phase of sporulation and
involves the following events:
● The prospore membrane closes
completely.

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 ● A thick wall starts forming around the
spores- This also contributes to the
maturation of the spores.
● Chromatic compaction in the nucleus of
the spores.
● Some of the organelles start
regenerating.

While given environments may contain

various nutrients, the lack of nitrogen and
other appropriate nutrient sources may
trigger sporulation of species like A.
nidulans and result in the growth of hyphae
and consequently conidiation.
Before spores start forming, special
reproductive hyphae known as
sporangiospores are produced.
Compared to sexual reproduction, asexual
reproduction is more common in fungi given
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that it allows for the rapid spread of the
organisms.
Some spores are produced in sporangium
(sac) through mitotic division (e.g. conidia).
Fungi spores vary in size and structure. The
differences allow them to be effectively
dispersed by a variety of agents including
wild, animals, and water.
Uses of Fungi:
Fungi are extremely important to the
ecosystem because they are one of the
major decomposers of organic material.
Decomposing organic material is how fungi
acquire energy. But fungi have other roles in
addition to being decomposers. How do
fungi help people? They are used to help
prepare food and beverages, and they have
many other uses.

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 Importance of Fungi in Human Use
● Yeasts are crucial for the fermentation
process that makes beer, wine, and
bread. Fermentation occurs in the
absence of oxygen and allows the first
step of cellular respiration, glycolysis, to
continue.
● Some fungi are used in the production of
soy sauce and tempeh, a source of
protein used in Southeast Asia.
● Fungi can produce antibiotics, such as
penicillin. Antibiotics are important
medicines that kill bacteria, and
penicillin was the first identified cure
against many deadly bacterial species.
Antibiotics only treat bacterial diseases;
they cannot be used to treat viral or
fungal diseases.

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● Mushrooms are fungi that are eaten by
people all over the globe.

Harmful Effects of Fungi:

1) Plant Diseases:
These are incited by different agents–
bacteria, viruses, cell-worms, and fungi;
of these the fungi cause the most severe
losses. They destroy field crops, forage
crops, fruit and nut crops, vegetable
crops, drug plant crops and ornamental
crops.

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2) Human Diseases:
Several fungi are responsible for
mycoses and tropical diseases of man
and animals.
3) Destruction of Timber and Timber
Products:
Several fungi like Polyporus,
Chloros-plenum, Penicillium divaricatum,
Fusarium negundi, Pori microspora,
Lentinus lepidens are responsible for
decay of standing trees, decay of felled
Timber, sap stains and decay of wood in
various uses.
4) Food Spoilage:
Besides microorganisms, members of
Mucorales (e.g. Rhizopus, Mucor), Yeast,
Moniliales, Aspergillus oryzae,
Penicillium digitatum are chief causes of
food spoilage. These are saprophytic

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 fungi which grow on the food articles
such as bread, jam, pickles, meat etc. For
instance, food items are spoiled by
yeasts, Aspergillus oryzae, Penicillium
digitatum etc. Dairy products are spoiled
by Aspergillus repens, Mucor, Penicillium,
Cladosporium, Oiduni lactis, etc.
5) Mushroom poisoning:
Deadly poisonous toadstools such as
Amanita sp and others have invariably
been mistaken for edible mushrooms and
have taken a toll of life. Mushroom
poisoning is known as mycetismus.

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 Experiment

Aim:
To study and ascertain the necessary
conditions required for the sporulation
of fungus on bread.

Materials Required:
Loafs of bread (4 in number), water (in a
sprayer), plastic bags (4), gloves, rubber
bands and marker pen.

Procedure:
● We have taken 4 loaves of bread in a
dish and with the help of a water
sprayer added a layer of moisture

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 (water) onto the surface of the bread
slices.
● We have used the plastic bags at our
disposal and inserted the bread into
the bags.
● We sealed the mouth of those plastic
bags with the help of rubber bands.
● Now we marked each of the prepared
bread loaf bags as A, B, C and D with
the help of marker pen.
● A was placed in a position inside our
house with the access to direct
sunlight.
● B was placed in a position with
access to diffused light.
● C was placed in a position with
access to no light at all (complete
darkness).

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 ● D was used as a control setup and
placed inside the refrigerator (cold
and dry condition).
● We note that all the places were in
hot and humid condition with the
exception of D.

Observing the bread samples at the end of

4th day.

A. Has clear signs B. Has little to no

of fungal growth signs of fungal
gro growth
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C. Shows few signs of D. Has no change
fungus.

Observations:
The setups have been observed after 96
hours (4th day) and 168 hours (7th day).
1.Day 0 (beginning of observation):
Setup looks normal, no change.
2.Day 4 (middle of observation):
A. Setup has clear signs of fungal
growth (white mycelium) with a slight
amount of visible spore formation

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 (dirty green and black color) albeit
the growth and spread is in less
amount.
B. Setup has very less to no signs of
fungal growth with just some strands
of visible white mycelium.
C. Setup has some signs of growth of
fungus with a few traces of spore
formation.
D. Setup has no change.

Observing bread samples at the end of 7th

day (end).

A. Sporulation has
increased, no increase in
mycelium.

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 B. Substantial fungal
growth and mycelium
covers the whole
surface.

C.Overwhelming
growth. Spores cover
the whole bread
surface.

D.
No Change.

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3. Day 7 (end of observation):
* All setups have clear signs of fungal
growth.
A. The amount of growth of fungi has
not increased. The sporulation has
increased. The amount of mycelium
remains the same.
B. The amount of growth of fungi has
substantially increased with the white
mycelium now covering even the walls
of the dish used. The amount of
spores and mycelium have both
increased substantially.
C. Overwhelming growth of spores and
mycelium is observed. Now the spores
cover the whole of the bread surface
and even the underside of the bread.
Only green color is visible in this
sample and the spores leave the
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 surface in the form of a thin mist
when disturbed.
D. No change is observed. (bread
became a little old).

Conclusion:
After 7 days, the bread loaf kept in the dark
(C )had the highest amount of growth of
fungi and sporulation. The diffused light
bread (B) has moderate growth of fungus
and the direct sunlight one (A) has the least
amount of growth. It must be noted that all
of the reads had substantial growth of
fungus. The exception is refrigerated bread
(D) which had no change or growth.

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 Precautions
1.Care has to be taken to supply an
adequate amount of water to the breads
and not overwater the loaves.
2.During observation, while handling the
loaves we must always wear gloves.
3.We always wash our hands after
handling the breads.

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 Conclusion
After completing the given investigatory
project and performing the experiment we
now have a brief idea as to what conditions
the fungi requires in order to grow and
thrive on common household items like
bread etc. We have concluded from the
experiment that the fungi requires a hot,
humid and dark environment to thrive
properly. We have also seen that fungi can
and will grow in direct sunlight and diffused
light as well though it does not grow in such
intensity as when kept in a dark
environment. We also have studied how to
prevent fungi from appearing on food
materials by storing them in the
refrigerator.

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 Bibliography
The following sources were used in the
making of this project:
1.NCERT Class XI Biology Textbook
2.Wikipedia
3.Brittanica.com
4.Modern’s ABC of Biology
5.ABD’s Laboratory Manual for Class XI.

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