Laboratory Exercise 7A: Diversity of Fungi

Biology II Laboratory BSC1011L Version: BJ AG NB DB AN SF BE Date:8March23__________________________________________________________________________________

Material is also covered in Chapters 31 of Campbell Biology, 12th Ed., Urry, Cain, Wasserman, Minorsky and Orr. Page numbers
refer to Chapter 6 (pages 135-148) of A Photographic Atlas for the Biology Laboratory, 8th edition.
Guides to supplementary pictures are in a small font. Examine the photos in the atlas if a concept is not clear.
Group Members: _____________________________________ ___________________________________________

_________________________________________ _________________________________________

KEY CONCEPTS:
1. Fungi are eukaryotes, and closely related to the animals.

2. Fungi are heterotrophs; they absorb their nutrition from dead organic matter
(saprophytes) or from living hosts (parasites).

3. The major groups of fungi differ in their asexual and sexual lifecycles, but the
groups we will examine have many similarities. In the sexual cycle,
plasmogamy or fusion of the cytoplasm occurs at a different time than
karyogamy, or fusion of the nuclei. Figure 1. Amanita phalloides
accounts for the majority of fatal
4. Fungi have diverse forms. Although there may be seven or more phyla, we will mushroom poisonings worldwide.
examine only four in lab. You may learn about the basal Cryptomycetes and http://en.wikipedia.org/wiki/Fungi

Microsporidians, and the Zoopagomycetes, that we will not examine here.

Main group Key traits that define the group
Cryptomycetes None in the laboratory, not covered in this lab.
Microsporidians None in the laboratory, not covered in this lab.
1 Chytrids: Some decomposers, but also many parasites of protists, other fungi,
plants and animals.
Contributed to global decline of amphibians.
Zoopagomycetes None in the laboratory, not covered in this lab.
2 Mucoromycetes Includes the fast-growing molds (bread, strawberries). Many are plant
Zygomycetes parasites or plant pathogens.
Plasmogamy produces the zygosporangium.
Glomeromycetes Form arbuscular mycorrhizae, structures critical for plant nutrition.
3 Ascomycetes: “sac fungi” Some yeasts, molds, morels, and truffles
In sexual reproduction, form saclike asci containing ascospores, often
in a fruiting body called an ascocarp.
4 Basidiomycetes: “club fungi” Mushrooms, puffballs, shelf fungi, forms that form mycorrhizae, and
parasitic rusts and smuts. Best groups for decomposing wood (lignin).
Form basidia or “little pedestals” which are club shaped (club fungi).

5. Lichens are a symbiotic association of a fungus with a photosynthetic partner (usually an alga or cyanobacterium)
that can produce food for the lichen from sunlight.
Lab 7A – Diversity of Fungi

SKILLS YOU SHOULD MASTER BY THE END OF THE LAB:

1. Recognizing the basic and life forms of fungi in four basic groups.
2. Recognizing the importance of the heterotrophic mode of nutrition to ecosystems.
3. Be able to understand the generalized life cycle of fungi (asexual cycle and sexual cycles).
4. Be able to identify the roles mitosis and meiosis play in the sexual life cycle, and the difference between
haploid, diploid and dikaryotic cells, and between plasmogamy and karyogamy.
5. Recognize that lichens are made up of fungi and algae and recognize their various forms.
6. Recognize the differences between symbiotic, parasitic, and commensal relationships

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7. VOCABULARY:
Mycology Zygosporangium Basidiospores
Hyphae Zygospore Gills
Septa Sporangium Chitin
Mycelium Sporangiophore Symbiosis
Stolon Mycorrhizae Mutualism
Rhizoids Ascus/ Asci Saprophytic
Plasmogamy Ascospores Parasitic
Karyogamy Conidia Commensal
Heterokaryotic Conidiophore Obligate
Dikaryotic Basidium/ Basidia Facultative

The material we will examine today:

Main group Types, comments Specimens you will examine
1. Chytrids One of the oldest lineages of None in lab Video clips
fungi Two species are considered the
cause of the current decline in
amphibians.

2. Mucoromycetes Formerly Zygomycetes- Rhizopus – Live culture (Demonstration)

Reproductive structure is a bread mold Prepared slide- Rhizopus
resistant zygosporangium. conjugation

Glomeromycetes- Form None in lab Note the arbuscular mycorrhizae

arbuscular mycorrhizae. and their importance

3. Ascomycetes Sac fungi (some yeasts, Peziza – Prepared slide-Peziza apothecium

molds, morels, and truffles). cup fungus
Penicillium Live culture
Sexual reproduction is in a Prepared slide-Penicillium conidia
sac-like structure called an Aspergillus Live culture
ascus. Prepared slide-Aspergillus
conidiophores

4. Basidiomycetes Club fungi (mushrooms, Coprinus Prepared slide – Coprinus

Lab 7A – Diversity of Fungi

puffballs, shelf fungi, rusts

and smuts, and some that
form mycorrhizae).
Sexual reproductive
structure is a basidium that
produces sexual spores.
Lichens are not a fungus, but a symbiotic association of a fungus and an alga.
Types, comments Specimens you will examine
Lichens Symbiotic association of an Crustose, Lab Specimens
algae and a fungus Fruticose, and Prepared slide-Lichen ascocarp
Foliose types Specimens on campus
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INTRODUCTION
Read through the material below and answer the following questions about fungi.
Mycology is the study of fungi. There are plenty to study, about 250,000 extant (not extinct) species.

Yeasts are unicellular forms:

Unicellular and free-living fungi are called yeasts. Some fungi have life stages that alternate between unicellular
and multicellular stages. The unicellular stages are referred to as yeast life stages. Most species that we consider
yeasts are in the Ascomycota, but single-celled forms (yeast forms) exist in the other groups as well.
Multicellular forms have many structures:

Multicellular fungi have bodies made

up of filaments, called hyphae. These
have exterior tubular cells walls. The
hyphae may or may not be divided into
smaller cells by septa, or cross walls
that allow the cell contents to pass
through them.

Hyphae secrete enzymes that break

down organic matter into nutrients, then
absorb them.

1. Which hypha in the picture is divided into smaller cells by septa? _________ (A, B, and/or C?)

2. Which hyphae are multi-nucleate (have many nuclei per cell)? __________ (A, B, and/or C?)

3. What are the small purple structures from which the hyphae grow? ___________________________________

The hyphae can be up in the air (arial

hyphae) and hold the sporangia or
spore forming structures. Other arial
hyphae can be horizontal connections,
much like branches, called stolon. The
fuzzy parts of bread mold you can see
are the arial hyphae.

The aerial hypha is made up of a

sporangiophore (stalk) and dark
spherical sporangium on top holding
the spores (see image below)

Hyphae can also penetrate living or dead

tissue (rhizoid hyphae). Rhizoids of
Lab 7A – Diversity of Fungi

bread mold penetrate the bread. A whole

network of interwoven hyphae that
penetrates another organism or structure
is called a mycelium. The mycelium is
the main body of the fungus.

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 The fruiting bodies (cap and stalk) are
temporary and spore producing
structures of fungi. The fruiting body of
a common mushroom is filled with
hyphae.

Some fungi have haustoria, or hyphae

that are modified into thin extensions
that can penetrate living cells and
absorb nutrients.

4. What are the filaments that make up all of the structures of fungi? _____________________________________
5. Name the structures:
• Hyphae that can grow up in the air and hold spore forming structures: ________________________________

• Hyphae that are horizontal making branch-like connections: ________________________________

• Hyphae that penetrate living or dead tissue: ____________________________________

• The network of hyphae that makes up the main body: _________________________________

• Spore producing structures: ____________________________________________

• Modified hyphae that penetrate living cells and absorb nutrients: ____________________________________

Nutrition:
• Fungi are sometimes called absorptive heterotrophs because the hyphae secrete enzymes that digest
material outside the cell. The hyphae then absorb the digested nutrients.
• Some fungi have a saprophytic mode of nutrition; they digest materials from dead or decaying organic
matter.
Cell structure:
• The cell walls of fungi are usually made of chitin, not cellulose (plants have cellulose). Chitin is found in
the exoskeletons of insects and crustaceans (animals). Strong cell walls allow the cells to absorb nutrients
from the environment without bursting from internal pressure.
Lab 7A – Diversity of Fungi

6. All fungi are heterotrophs. What is a heterotroph?

7 Most fungi are saprobes; they have saprophytic mode of nutrition. What is a saprophytic mode of nutrition?

4
Examine Figure 2: Generalized Fungal Life Cycle. Read through the following material as you examine the
generalized life cycle. There is great variety in the reproductive cycles of fungi, but you will only be responsible for
knowing the generalized life cycle in Figure 2 on the following page.

Asexual reproduction: All stages of asexual reproduction are haploid (1n) and undergo mitosis.
• Spore production within sporangia, conidiophores, or other spore-producing structures.
• Budding or fragmentation. The growing bud or the broken off fragments of mycelia detach and grow into new
mycelia. The mycelia that are produced by asexual reproduction are genetically identical.

Sexual reproduction: Strains of mycelia that are genetically distinct may reproduce sexually, even though they do not
have male and female forms. Mycelia produce pheromones, from which mycelia can tell if they belong to the same strain
or to different mating types. If the mating types differ the mycelia may grow together and their cells fuse.
1. Plasmogamy - the first stage of cell fusion, where the cytoplasm of both cells fuses. Following plasmogamy,
the cells contain two nuclei and are called heterokaryotic (n+n).
2. Karyogamy - the nuclei fuse to become diploid (2n) zygotes.
3. The zygote undergoes meiosis to produce haploid spores (zygospores) which grow into new mycelia. However,
the spores produced by karyogamy and meiosis have new combinations of genes from the two mating types and
produce genetically differentiated mycelia. In some cases, fungi remain dikaryotic (have two distinct nuclei)
after plasmogamy and form mycelia without undergoing meiosis. These fungi may remain dikaryotic for hours,
days or centuries, before going through karyogamy.

n: __________
______

n: _____
n: _____ ______
______
n: _____
______
n: _____
______

n: _____
zygospores ______ Lab 7A – Diversity of Fungi
Figure 2: Generalized Fungi life cycle (Figure 31.5 of your text book by Campbell and Reece).

Identify the number of chromosomes at each step above. If you have trouble, read the following section.
• Most of the life stage of the fungus is haploid (1n). Haploid cells (1n) replicate by mitosis, resulting in cells with
identical number of chromosomes (1n). Haploid stages include the mycelium (main body of a multicellular fungus),
asexual reproduction of spores, production of spores from the zygote undergoing meiosis, and in life stages up to
plasmogamy. Both asexual and sexual reproduction involves the release of spores (1n), which can travel long
distances in the wind, and survive until there is enough water to germinate and produce mycelia.
• The only diploid stage (2n) follows karyogamy in the sexual reproductive cycle, where nuclei fuse to form the 2n
zygote. In the heterokaryotic state the cells have multiple nuclei, but the nuclei are not fused, they are “n+n”. Be
sure to identify the diploid and heterokaryotic stages on the figure with “2n” or “n+n”.
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Examining the Organisms:
1. Chytrids: Phylum Chytridiomycota. Figures 6.2-3. There are about 1000 species. Some are decomposers,
and some are mutualists, breaking down plant matter in the digestive tracts of cattle.
Others are parasites of protists, other fungi, plants and animals.
The loss of the Golden Toad is one of several examples of the devastation caused by Batrachochytrium
dendrobatidiscan. Part 1: http://www.youtube.com/watch?v=YIw5roHzwPE&feature=results_video&playnext=1&list=PL71B8A9972187AD97 8:09 min
Part 2: http://www.youtube.com/watch?v=kJdhNy6OPrM&feature=related 8:44 min

The current leading hypothesis for the

decline of many amphibians (frogs,
salamanders, caecilians), is infection by the
chytrid fungi. One is Batrachochytrium
dendrobatidis, “Bd” for short (Atlas Figure
6.3). Because chytrids are ubiquitous among
the habitats of amphibians (they are found
mostly in freshwater lakes and soils), it is
unclear how humans can assist in the
conservation efforts as eradication of the
fungus is not possible.
http://www.youtube.com/watch?v=ZWHibA
Q0Sso

http://theworldofrogs.weebly.com/chytrid-fungus.html

The small circular structures are zoosporangia of B. dendrobatidis. scale

bars = 30 um

A- growing on a freshwater arthropod

B- growing on an alga

The zoosporangia are the spore cases in which the zoospores develop.
The zoospore is an asexual spore that is motile and flagellated (it uses a
flagellum for locomotion).
Lab 7A – Diversity of Fungi

https://en.wikipedia.org/wiki/Batrachochytrium_dendrobatidis

6
 2. Mucoromycetes: Includes the:
• former phylum Zygomycota (conjugation fungi; Figures 6.4-6.10)
• Glomeromycetes.
Zygomycota form the resistant spherical spores in the zygosporangia during sexual reproduction. They are mostly
terrestrial in habitat, living in soil or on decaying plant or animal material.
Glomeromycetes are best known for their arbuscular mycorrhizae, the branched hyphae forming an arbuscular
that assist plants in getting critical nutrients.

Examine the life cycle of Rhizopus (Figure 6.4). Identify the stages of the generalized life cycle as it is found in
Rhizopus.

Asexual cycle-
hyphae feed and grow
on bread. Note that the
hyphae have been
modified into rhizoids
which can attach to a
substrate, and stolons
which connect to one
another.
• Rhizoids typically
grow into the
bread.
• Stolons can be
seen on the bread,
and
sporangiophores
rise from the
stolons.
• Spores
(sometimes called
sporangiospores
to differentiate
from zygospores)
are released from
sporangia
(singular-
sporangium).

Sexual Reproduction (larger cycle above) Mating types (+,-) touch each other, forming gametangia within
Lab 7A – Diversity of Fungi
each strain. The nuclei begin to differentiate and prepare for plasmogamy.
• Plasmogamy occurs when the walls between the gametangia break down forming the zygosporangium. It is
heterokaryotic (n+n) Identify where Plasmogamy occurs on the image above.
• Karyogamy occurs within the zygosporangium when the haploid nuclei fuse and form the 2n nuclei. Identify
where Karyogamy occurs in the image above.
• When conditions are right, the 2n nuclei undergo meiosis and haploid (1n) spores are formed.
The spores germinate to form hyphae.

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1. Demonstration Bread Mold in the hood: Observe the bread mold (a piece of moldy bread in a dish) under
the dissecting scope in the hood. Note that you may see several species differentiated by having mycelia or sporangia of
different forms and colors.

2. Demonstration: View the pure culture of Rhizopus (live culture on a plate) under the dissecting scope.

Do you see sporangiophores with sporangia? _______ (the dark spots in the photo above) Use Atlas figure 6.4-6.10.
The fuzzy parts of the photo are the drops of water on the lid of the petri dish.
Lab 7A – Diversity of Fungi

Are the sporangia diploid or haploid? ____________________________

What is the only structure that is diploid? __________________________________

What is the difference between dikaryotic and diploid?

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3. Look at the Prepared Slide: Rhizopus Conjugation

The zygosporangium is the deep red color. See also Atlas figure 6.10.

Low power: 100X High power: 400X

Are the nuclei in the zygosporangium initially 1n or 2n?________

Once the nuclei pair, but before they fuse they are: _________________

Once the nuclei fuse they are (1n or 2n) ______________ .

The nuclei then undergo meiosis to form spores. The spores are (1n or 2n) _____________.

Glomeromycetes: Glomeromycota have critical

ecological functions. Nearly all form arbuscular
mycorrhizae. The branched hyphae form an arbuscule (a
highly branched tree-like structure). These fungi assist the
plant in getting critical nutrients such as phosphorus,
sulfur, nitrogen and other micronutrients, from the soil.
http://www.youtube.com/watch?v=PiZHFaQdcX8

This association may have played an important role in the

colonization of land by plants, allowing plants to survive
in regions with low nutrients. Many plant species (80% of
plant families) have mutualistic relationships with these
fungi and ecologists are using this knowledge to advance
our understanding of ecosystem management and
agriculture.
Lab 7A – Diversity of Fungi

Figure 3. Flax root cells with arbuscules

http://en.wikipedia.org/wiki/Arbuscle

What percentage of plant families have mutualistic relationships with Glomeromycete fungi? ______

Why are Glomeromycetes so important to humans?

9
3. Ascomycetes: Phylum Ascomycota, or Sac Fungi. These include many yeasts, and the molds, morels, and
truffles (Figures 6.1-6.26).
Ascomycetes are an extremely significant and successful group of organisms. Familiar examples of sac fungi include:
Baker’s yeast (Figure 6.12)
cup fungi (Figure 6.16)
morels (Figures 5.16)
truffles
Dead Man's Fingers.
• During sexual reproduction, Ascomycota produce nonmotile spores (ascospores) in a distinctive type of microscopic
cell called an ascus.
• In some ascomycetes, during asexual reproduction, asexual spores called conidia are created. Conidia are not formed
in sporangia, but rather at the tips of specialized hyphae called conidiophores. Conidia may also fuse with hyphae of
an opposite mating type in sexual reproduction.

Examine Figure 6.11. Life cycle of an ascomycete. Label plasmogamy and karyogamy. Find the ascus
containing the ascospores.

Lab 7A – Diversity of Fungi

In the asexual cycle, ascomycetes reproduce by forming spores called ascospores which grow into hyphae of different
mating types. Label plasmogamy and karyogamy.
In the sexual cycle, hyphae from two mating strains touch.
• Plasmogamy occurs when the antheridia and ascogonia of the different strains fuse.
• Karyogamy occurs when the haploid nuclei fuse to become 2n.
• Haploid ascospores are produced through meiosis.
The ascospores can reproduce by mitosis and grow from 4 cells to a typical 8 cells while still encased in the ascus (Figure
6.18). Each ascospore can germinate to form a new mycelium.
10
Examine prepared slides and demonstrations:

1. Cup Fungus - Prepared Slide—Peziza asci (CS)

(Figures 6.16-6.18)

This slide is a cross section of the asci from the lining of the
ascocarp of a cup fungus.
A. Are the asci (ascus=singular) sexual or asexual

reproductive structures? _______________________

B. How many ascospores do you see in each ascus? _______

C. Are the ascospores diploid or haploid? _______________

High Power:400X

The term “yeast” does not identify a true evolutionary group but includes the many unicellular fungi. Because it is
difficult to identify the true evolutionary group that a yeast belongs to, they are considered “imperfect fungi” until they
can be correctly classified.

Although the term yeast refers to unicellular forms, some species with yeast forms may become multicellular through the
formation of a string of connected budding cells known as pseud hyphae, or true hyphae as seen in most molds. Most
yeasts reproduce asexually by budding, although a few do so by binary fission.

2. Yeast (Saccharomyces) (Figure 6.12): Within the

Ascomycota, there are approximately 1,500 described species
of yeast. The yeast species Saccharomyces cerevisiae has
been used in baking and fermenting alcoholic beverages for
thousands of years. It is also extremely important as a model
organism in modern cell biology research and is the most
thoroughly researched eukaryotic microorganism.

This is the species of yeast used make bread rise.

Lab 7A – Diversity of Fungi

High Power 600X

11
3. Penicillium and Aspergillus: Examine Penicillium from a prepared slide (in blue box) or live culture, and
prepare a wet mount of Aspergillus from the live culture stored in the hood)

Examine the Penicillium and label the conidiophores and High power: 400 or 600X
attached conidia. Look also at figure 6.22

Conidiophores are made up of chains of bead-like

conidia, which are modified cells growing from the
hyphae.

Low power:

Examine the Aspergillus and label the conidiophores and High power: 400X
attached conidia. Look also at figures 6.23-6.26.
Low power:

Very high power – see the conidiophores (stalks that

produce the conidia) and conidia (asexual spores)

Lab 7A – Diversity of Fungi

12
4. Basidiomycetes: Phylum Basidiomycota- Club fungi (mushrooms, toadstools, rusts and smuts, figures
6.27-5.40).

D Examine Figure 6.28- Basidiomycota are filamentous fungi composed of hyphae (except for those forming
yeasts) and reproduce sexually via the formation of specialized club-shaped end cells called basidia (singular-
basidium). Basidia normally have external spores (usually four). Some Basidiomycota reproduce asexually also. In
figure 6.28 locate the basidia, the basidiospores, and the now familiar dikaryotic stage following plasmogamy, and
the diploid stage following karyogamy. Locate the gills on the undersurface of the cap (pileus). The fruiting body
(e. g. mushroom) is the basidiocarp and develops from the long-lived dikaryotic mycelium.

The Basidiomycota include puffballs (Fig 6.30), stinkhorns, bracket fungi, jelly fungi, chanterelles, and earth stars.
Many Amanita species (Figure 6.27) are highly poisonous and one bite can be fatal.
In contrast, Agaricus bisporus (Figure 6.29) is cultivated for human consumption.

Life cycle of basidiomycetes.

https://www.sciencedirect.com/topics/agricultural-and-biological-sciences/basidiomycota

Is the main body you call a “mushroom” haploid, diploid or dikaryotic? _____________________________
Lab 7A – Diversity of Fungi

Which part of the mushroom is diploid? ____________________________________

Basidiospores are formed thought meiosis at the tip of the basidium. This results in the spores being (haploid, diploid or

dikaryotic)? __________________________

Identify where plasmogamy occurs (where do monokaryotic cells become dikaryotic)?

Identify where karyogamy occurs (where do the nuclei fuse)?

13
1. Examine the prepared slide: Coprinus (CS). Use Figures 6.27(cap) and Figure 6.32 (the gills at 40X and 430X) as guide

A. The gills of the Coprinus as seen through your microscope, basidia and the basidiospores.

Cross section of mushroom cap. You an see Edges of gills with

the many gills. 40X many basidia. 100X One basidium with basidiospores at tip
600X

Lab 7A – Diversity of Fungi

14
 Lichens (Figures 6.41-6.50) are a symbiotic association of a fungus (usually an ascomycete) with a
photosynthetic partner (usually either green alga or cyanobacterium) that can produce food for the lichen from
sunlight. Symbiotic species live in a close, long-term, and often dependent, associations.

Some symbiotic associations are obligate, in that the species cannot live without each other; other associations are
facultative, where the relationship can be beneficial to either organism, but it is not essential. Some lichens are given
scientific names as if they were single organisms, because the two species are so dependent on each other.

Lichen are mutualists, because both the algae and fungi benefit from the association. Remember, in parasitic
relationships only one species benefits at the expense of the other; in commensal relationships, one organism is
benefited, but the other is neither helped nor harmed.

In lichens, a mass of fungal hyphae surrounds the algal cells, often enclosing them within complex fungal tissues
unique to lichen associations; however, the algal cells are never enclosed inside the fungal cells themselves (Figure
6.42). Lichens often reproduce asexually (Figure 6.42) by releasing small soredia, or particles that contain both algal and
fungal cells. Both fungus and algae can reproduce sexually by the mechanisms unique to their phylum.

Air pollution often destroys lichen, perhaps because lichen absorb airborne nutrients and minerals efficiently
because they are dependent on those particles for life.

Lichen are a symbiotic association of ___________________________ and ___________________________________

Symbiotic means: _________________________________________________________________________________

Obligate symbiosis means: __________________________________________________________________________

Facultative symbiosis means: ________________________________________________________________________

Lichen are mutualistic and this means: _________________________________________________________________

Parasitic means: ___________________________________________________________________________________

Commensal means: ________________________________________________________________________________

Would you be more likely to find lichens in urban or rural situations? ________________________________

Why? ___________________________________________________________________________________________

Lab 7A – Diversity of Fungi

Crustose lichen (crust-like; see Figures 6.41a, 6.47) grow on

hard substrates such as rocks or bark, and usually lie flat
on the substrate.

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 Foliose lichen (leaf-like; Figures 4.41b, 6.46, 6.48, 6.50) also
grow on firm substrates, but portions of the lichen form
structures that sometimes look like sticks, leaves, or the
caps of mushrooms.

Fruticose lichen (shrub-like; Figures 6.41c, 6.49, 6.50) often

grow away from the substrate like shrubs and may have
stalks whose tips are the sexually reproducing fungi.

Name the three forms of lichen:

Lab 7A – Diversity of Fungi

1. View the lichens on display, on prepared slides and examine the lichen in Figures 6.41 and 6.46- 6.50.

16