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Lab 09 Protists and Fungi

Roadmap to the rest of this semesters labs
We will be traveling through the worlds diverse life forms using evolutionary trees as our roadmaps. To
do so, we will follow the branches of a given phylogeny (like the one in the taxonomy lab manual). The
different branches on a phylogeny indicate groups that share certain characteristics. Characteristics that
are kept probably helped them survive (or at least didnt hurt them) and are therefore adaptations.
Adaptations are traits that are selected for. These adaptations typically evolved at a single point sometime
in the groups history. So, during these labs, as we travel along a branch, we will add key adaptations
(really important adaptations that split and define groups and allow them to radiate) to the branch, thus
narrowing down and providing specific definitions of the taxonomic categories beyond that point on the
branch.

DOMAIN EUKARYOTA
The first key adaptations on our journey past prokaryotes are membrane-bound organelles and the
nucleus. Note: this means that prokaryotes do not have these. They evolved in another lineage that split
from them.

Membrane-bound organelles
Organelles are little compartments inside cells that are
specialized for certain functions. You might remember
the types and functions of many organelles (like the
nucleus, Golgi apparatus, lysosome, etc). The evolution
of organelles is uncertain. It could have been through a
close relationship between two prokaryotes in which one
lived inside another (endosymbiosis). They may have
relied on each other so closely that they began losing their
own independence eventually fusing into one cell. This is
most certainly how mitochondria and chloroplasts, both
are organelles, evolved.


Nucleus
The nucleus is the organelle that houses the chromosomes and therefore the vast majority of the genetic
material. All eukaryotic cells have nuclei at some point, although some cells lose theirs during
development (like red blood cells). The reason the nucleus evolved is still a mystery, although several
hypotheses have been proposed. It could have been similar to the endosymbiosis hypothesis mentioned
above. At any rate, all eukaryotes today have nuclei and nuclei are essential for their survival.

Examining the Protists
The Kingdom Protista is an incredibly diverse amalgam of many different forms of life. There is no
single trait that unifies them or that separates them from the other eukaryotic kingdoms: Plantae, Fungi,
and Animalia. Protists are not monophyletic by any means and their evolutionary relationships have not
been completely sorted out; some are closely related to animals, some to plants, and some to each other
(see the phylogeny in the taxonomy lab). Some taxonomists now split this kingdom into many different
kingdoms.

Most protists are made up of unicellular eukaryotes, but there are notable exceptions, such as the giant
brown algae (kelp). Protists have many niches, some living in aquatic environments, some live in soil,
while others are parasites living in the blood stream of other organisms. Today you will be mostly
examining the group formerly known as protozoans, which are heterotrophic and rely on engulfing other
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organisms for a food source, and so were thought to be more closely related to animals than plants. Later
on, well explore the group of protists formerly called algae, which are autotrophic organisms, capable of
making their own food through either photosynthesis or chemosynthesis, and were thought to be more
closely related to plants (but again these relationships are not well determined. For example, see the tree
in the taxonomy manual and find brown algae).

Today in lab, we will have living and preserved example organisms from the protists. Look at these using
the 40x objective. Once again, draw what you see and record any extra information. You are
responsible for distinguishing between species during your exam. Here are some important specimens to
know. Their group names are based on molecular similarity and are given just as a means of organization
(no need to memorize the group names).

I. Chromalveolata
A. Alveolata
Ciliates move via many hair-like projections from the cell membrane called cilia.
o Paramecium- commonly found in water feeding on microorganisms like
bacteria. Despite being one cell, they have been shown to possess the
ability to learn and possibly communicate with each other through
radiation.
- See slides marked Plasmodium caudatum wm
- There may be other ciliates in lab today, like living Stentor and Vorticella.
Make a wet mount of these. It is fun to watch these critters move and feed.

Dinoflagellata is a diverse group that contains predators, photosynthesizers, and
endosymbionts. Most live in marine ecosystems and swim with two long flagella. Some
dinoflagellates are covered in plates of armor made of cellulose or silica.
o Those that are photosynthetic play a major role in marine ecosystems as primary
producers (i.e. they are the bottom of the food chain.) Some live inside coral
animals and help them form coral reefs (aka zooanthellae).
o Dinoflagellates can cause red tides when their populations bloom.
- See slides marked Dinoflagellates and Ceratium wm

Plasmodium- this mostly immobile, single-celled parasite causes malaria- one of the most
common diseases in humans worldwide, killing around 781,000 people each year ( thats
2.23% of human deaths!). It spends some part of its life in female mosquitoes, which
inject the protist into a humans bloodstream when it bites. The protist then continues its
develop inside the human red blood cell.
- See slides marked Plasmodium falciparum
smear and Plasmodium vivax smear

B. Heterokontophyta (aka Stramenopiles)
Diatoms are perhaps the most common life forms that make up
phytoplankton, the microscopic photosynthetic organisms that
form a greenish layer on the surface of the ocean. As such,
they form a critical first step in the food chain; they make food
from sunlight, are eaten by bigger things, which are then eaten
by bigger things, and so on. They make a hard cell wall of
silica (called a frustule). Silica is sand or glass and is the most
common mineral in earths crust. The shells of dead diatoms
make up part of the ocean floor and are harvested for
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diatomaceous earth, which is used as a filter, abrasive (as in toothpaste), and absorbent (as
in cat litter).
- See slides marked Diatoms: freshwater mixed wm, frustules, and
comparison pennate and central wm

Brown algae is unique among the protists from todays lab in that they are multicellular
and can even form giants growing to over 45 m (150 ft). Better known as kelp, a type of
seaweed, these brown algae can form huge underwater forests that serve as homes to many
famous sea creatures, like sea otters and sharks. In lab today, we may have several forms of
brown algae, including slides and specimens in jars or mounted.
- Look for these names: Fucus, Laminaria sporangia, Vaucheria
geminate,and Sargassum



II. Excavata
A. Euglenozoa
Euglena-These little critters are commonly found in pond water.
They are often called algae, but are heterotrophic and
photosynthetic, which is pretty cool. They move via one long
flagellum, which you may or may not be able to see on the slides.
- See slides marked Euglena wm and Euglena
flagella

B. Metamonada
Trichomonas vaginalis is an internal parasite that often lives inside the human urogenital
tract. Infections in woman usually cause inflammation of the cervix and potentially birth
defects, whereas infections in men are usually asymptomatic. It
is transmitted through sexual contact. More than 160 million
people worldwide are annually infected by this protozoan.
Lesions found in the fossil jaws of Tyrannosaurus rex were likely
caused by a parasite similar to Trichomonas gallinae.
- See slides marked Trichomonas vaginalis smear



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C. Sarcomastigophora
Trypanosoma look like little worms, but they are still a single cell and live among human
red blood cells.
o T. brucei (aka T. gambiense) is transmitted in the spit of the tsetse fly and causes
sleeping sickness (which causes fever, headaches, joint pains, and leads to immune
deficiencies). It is estimated that 50,000 to 70,000 people are currently infected,
mostly in sub-Saharan Africa.

o T. cruzi is transmitted by an assassin bug and causes Chagas disease, which can
affect the nervous system, digestive system, and heart. It is predominantly found
in Central and South America. Since it has a chronic stage, this protist might be
why Darwin was sick in his old age.


Giardia lamblia is by far the cutest single-celled organism, but unfortunately it is passed
from the intestines to fecal matter in the water supply; if ingested, it can cause hikers
diarrhea.
- See slides marked Giardia lamblia trophozoite

III. Rhizaria
The Rhizariaare typically single-celled marine creatures that form intricate shells made of
calcium carbonate or sulfates. They float or move by extending parts of their cell through the
shell in skinny projections called pseudopodia. When they build their shells, they remove
atmospheric carbon (after it falls into the ocean during rains). When they die, these shells drift to
the ocean floor and become sediment, which builds up over hundreds of years (see the white
cliffs of dover). As an added bonus, these creatures protect our atmosphere by reducing the
greenhouse effect. Unfortunately, we like to put way too much carbon in the atmosphere and
change in the pH of the ocean, so they can no longer keep up.
A. Foraminiferans B. Radiolarians


IV. Amoebozoans
A. Amoeba are single-celled and formerly aligned with the
protists, but they are actually more closely related to animals
and fungi (in the group Unikonta). Amoeba move by
extending part of the cell forming pseudopodia (fake feet)
and then drawing the rest of the cell in that direction. They
feed similarly by extending part of its body around the food,
eventually engulfing it.
- See slides marked Amoeba proteus wm
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KINGDOM FUNGI Is there a fungus among us?
The short answer is yes, and the long answer is youre related to one. For a long time, fungi were
considered plants. I mean shoot, they dont move and they look like little plants, right? Well, fungi are
far from plants. Molecular studies have shown that fungi (plural, the singular form is fungus) are more
closely related to animals than to plants [though they are not closely related to any extant (still living)
group of organisms]. (I dont know why everyone keeps putting them in book chapters about plants.)
They do not have chlorophyll, they are not photosynthetic, they are not made of cellulose, and are instead
heterotrophic, meaning that they must get their energy and carbon from other organisms, just like
animals do. Unlike animals, they feed by digesting their food outside of their bodies, then absorb the pre-
digested material. They do have cell walls made of chitin, a structural protein that we will see again in
the arthropods. That is why fungi are on a different branch of the phylogeny of life than plants and are
instead lumped with slime molds and animals.

Besides that, all of us have fungi living on or in our bodies. In fact, every breath you breathe probably
includes some fungal spores. Take a moment to let that sink inInhaleexhaleThough some fungi
can cause human disease and hurt agricultural crops as well as native
vegetation, fungi play a very important ecological role as a decomposer.
Without fungi, the forest floor would be covered in woody debris. Fungi are
one of the only organisms that possess enzymes capable of breaking down
lignin (one of the major components of wood) into smaller molecules usable
by other organisms. Their presence in an ecosystem is essential to nutrient
cycling and the sustainability of that ecosystem. Fungi either live off dead
organic material (in which case they are called saprobes) or they live as
symbionts (a partnership with another living organisms). The symbiosis can
either be parasitic, in which the fungus feeds on another organism to the
detriment of that organism, or a mutualistic, in which both partners benefit.

Fungi are amazingly diverse; ranging from single-celled to
multicellular; asexually reproducing, sexually reproducing, or
both, and more. Therefore, it is hard to make generalities, but Ill
try. The anatomy of a multicellular fungus consists primarily of
filamentous hyphae. They are strings of cells separated by walls
called septa, which are rarely a complete barrier from one cell to
the next. Its the hyphaes job to spread out over an area and
absorb nutrients, energy, and carbon. They then share these
nutrients with the rest of the fungus. Since fungi feed through
absorption, they need a lot of surface area per volume. So, which
would be more efficient at absorption, short fat hyphae or long
skinny hyphae? A large number of hyphae grouped together make
up the body (thallus) of a fungus, called a mycelium.

Use the slides marked Aspergillus or the whole bread mold specimens to see the hyphae gathering
together to form mycelia.


The reproductive structures, such as the aboveground structures we often identify with fungi (the
mushroom caps, for instance), are used to separate the fungi into different phyla. Most of the time, the
fungus stays happily underground growing hyphae.

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Fungi have very complex life cycles that often involve both sexual and asexual reproduction. (By sexual
reproduction, I mean that organisms must share genes. Asexual reproduction means that an individual
can reproduce without combining genes from another individual. These terms have little to do with sexual
intercourse; some asexual animals actually have sex, but do not use the sperm and some sexual organisms
never get near each other.) When fungi reproduce sexually, they dont necessarily have a male and
female fungus, rather two fungi of different types must come together to share genes.



There are four different phyla that make up the majority of fungi based on their reproductive structures
(see list below). The rest of the fungi are lumped into the group either known as Imperfect fungi or
Deuteromycetes, because scientists have not observed their mode of sexual reproduction. We will focus
mainly on the zygomycota, ascomycota, and basidiomycota in this lab.


Note: The term mold is an informal term meaning rapidly growing fungus that reproduces asexually by
producing spores. The term yeast is a single-celled fungus that reproduces asexually by cell division or
budding. Both molds and yeasts are found in many phyla.
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Today, you will examine the examples from most of these phyla in the lab. Make sure you note the
different structures. There are dissection microscopes available so that you may see the exterior
structures with some magnification.



Phylum Chytridiomycota
Chytrids are among the oldest known fungi. Their gametes still have flagella. There
arent that many well known, but they seem to be one of the causes currently killing
amphibians, a phenomenon known as amphibian decline. The frog on the right is
missing a limb due to developmental issues caused by a chytridiomycote. (Over 30% of
amphibian species are threatened with extinction from various causes, including this
fungus.)
- See slides marked Allomyces





Phylum Zygomycota
Zygomycetes include the common black bread molds and fruit molds that often spoil your
lunches. They reproduce sexually when two nearby hyphae grow toward each other (lead
by pheromones) until they touch. They then fuse together and form a tough structure
known as a zygosporangium. This structure holds the genes from
both parent cells until conditions are right (like moisture returns).
When things are lookin good, the cells will fuse their nuclei (fertilization) to form
a sporangiuma stem-like projection in which spores are made. The mold slides
show the sporangia of mold.

- You can see examples of this phylum and the zygosporangium in the slides
marked Rhizopus conjugation. These slides show where two types of
hyphae have merged to form a zygosporangium. This is the dark lump that
connects two cells.




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Phylum Ascomycota Sac fungi
The Ascomycetes produce a saclike structure that produces spores (hence the
common name) called an ascus hence the taxonomic name). These include several
fungi that you may have heard of (or imbibed):

Some yeasts turn glucose into ethanol and CO
2
, which we use in bread making (the ethanol is burned
off, the CO
2
causes the bread to rise) and beer and wine making (the yeast turn the sugar from the
hops or grapes into booze and the CO
2
makes it bubbly).
- We have slides of two types of yeast which are not closely related and
are separated by the way they reproduce: budding yeast (Saccharomyces)
and fission yeast (Schizosaccharomyces)

Cup fungi form a large mushroom-like
mycelia (called an ascocarp) that is
sometimes cup shaped. Morels and truffles,
rare and very expensive delicacies, are cup
fungi.







There are many molds that are ascomycetes. For example, there are
several plant pathogens, including the ones that cause Dutch elms disease
and chesnut blight, which has nearly wiped out the American chestnut.
Once very common in the US, there are no mature chestnuts anymore
because this fungus knocks them back before they get old. Tinea is a
human pathogen that causes ringworm, jock itch, and athletes foot. To
make up for some of these assaults, Penicillin, the mold that produces the
antibiotic penicillin is an ascomycete.

We have several ascomycote specimens and slides in lab today,
including
- Peziza, saprophytic fungi that feed on wood
- Penicillium, the maker of penicillin antibiotics
- Aspergillus, which commonly grows on bread



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Phylum Basidiomycota Club fungi
The basidiomycetes are the fungi that you think of when you think of fungi (as you do). They form a
large basidiocarp or club (the above ground mushroom that youve seen several times). If you look
closely enough at a mushroom you can see that the solid cap is really formed from filamentous hyphae
bunched together into a mycelium. Im not sure if we can do this in the lab, but you can try.

The mushroom club is really a reproductive structure. The gills on the underside of the cap (lamellae) are
full of thousands of basidia (singular: basidium). The basidia produce spores (or basidiospores, I swear
its like an old comic book where everyday objects are given the characters theme name, like Spider-
mans Spider-car and Spider-dog and Spider-curtains and Spider-toilet). These spores are released to
form new hyphae elsewhere.











Examples of basidiomyces include:
- Mushrooms- e.g., Agaricus bisporus (tasty pizza mushroom) and
Coprinus mushroom

- Puffballs- Full of spores that shoot out when the puff is broken and
squeezed

- Bracket or shelf fungi- found on trees in the forest

- Rusts and smuts- serious plant pathogens that often
damage crops. See slides of Puccinia (Wheat rust) and
Ustilago (corn smut).




uffball
Shelf fungl
Corn smuL
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Lichen
Remember when I said that fungi do not have chlorophyll, they are not
photosynthetic, they cannot make their own energy, and are instead
heterotrophic, meaning that they must get their energy and carbon from other
organisms? Good times. Well, some fungi have figured out a way to get
around this always-needing-a-source-of-nutrients problem. They carry their
own bundles of photosynthetic goodness. Lichens are a symbiotic relationship
between a fungus and a green alga. Green algae are mostly single-celled,
autotrophs that photosynthesize light energy into sugars using chloroplasts?
That is what the fungus in a lichen is taking advantage of. By carrying its own food processing plant, a
single-celled alga, they can live almost anywhere they land, like the side of a building or a rock. In turn,
the algae receive a nice home and an attachment point on an otherwise hot, dry rock or tree trunk.
Therefore, this symbiosis is mutualistic- both parties benefit. Lichens
are very common and often form the first colonies of living organisms
in new barren landscapes. They are pioneers.

We have slides of this too. Look for the single-celled algae wrapped
up snugly in the fungus hyphae like a kid with an ear infection being
comforted by his mom on Christmas Eve.









As a side note: NEVER NEVER NEVER eat
mushrooms in the wild unless you are a trained
mycologist! You cannot tell the edible ones from the
poisonous ones. And sometimes they look the same
during different parts of the country or at different times
of the year. The cartoon by David Shrigley is meant as a
joke, but it is pretty accurate.


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"#$% &&'( %)* '+ ,-./01/1 )23 45260 7581/0.21 name_____________________


1. Explain the endosymbiont theory as a potential explanation for the evolution of eukaryotes.









2. Define the major feeding strategies utilized by the groups in this lab and name a specific type of
either a protist or fungus that does each:
A. Autotrophic


B. Heterotrophic


C. Saprotrophic


D. Parasitic


3. Draw a Paramecium. With what structures does it move?








4. Draw Trypanosoma in its natural habitat and label the protists and red blood cells.





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5. What is a red tide? (and I dont mean an elephant.)









6. Malaria is a devastating disease that kills millions of humans worldwide. About 250 children in
Africa die of the disease every hour. It is caused by a protozoan that is inserted into humans via
mosquito spit. On a physiological level, how does Plasmodium cause malaria? In other words, what
does the Plasmodium do to the human body to cause the symptoms we call malaria? In a separate part
of your answer, list the symptoms.



















7. Name two traits that fungi share with
animals, but not plants.
A.



B.


8. Name two traits that fungi have and animals
dont.
A.



B.



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9. What is the main purpose of the mushroom structure?




10. What are exoenzymes and how are they used by fungi?





11. Draw what bread mold looks like under a scope. Label the hyphae and the fruiting bodies.














12. Lichen is a symbiosis between
A. A plant and a fungus
B. A fungus and an alga
C. A Simon and a Garfunkel
D. A tapeworm and an intestine



13. Another important ecological role that fungi play is in the form of mychorrhizae. (We have slides of
these too.)
A. Mychorrhizae are a relationship between what two types of organisms?


B. What type of relationship are they in?


C. Explain what each member of the relationship contributes to the other member.