BIOL227 - Lab 3 - Protists F2024

Lab 3: The diversity of protists

Written by: Dr. Catherine Calogeropoulos ©

Edited by Dr. Amandeep Glory and Dr. Andrew Wieczorek

General information about protists

While protists are (mostly) single celled, they have more structural, morphological and organellar
diversity than any other group of eukaryotes. To support this diversity, their organelles take on more
diverse roles than those of multicellular eukaryotes. While an important part of this lab is to identify the
unique features of each supergroup, we are mostly focused on identifying and appreciating the structural,
morphological and organellar diversity of protist clades within each supergroup.

You will also identify and explain the principal modes of nutrition, locomotion and reproduction
observed in protistan clades. Protists include important photoautotrophs (e.g. euglena, diatoms, brown
algae, dinoflagellates, volvox and spirogyra) which account for roughly 30% of the net primary
productivity of our planet. Such protistan photoautotrophs are loosely called algae. Protistan
heterotrophs are called protozoa. The protozoans are further categorized based on how they eat:
ingesting other organisms (these are the phagotrophs, e.g. paramecium, amoebas), feeding on host cells
(these are the parasites, e.g. Giardia, Trichonomas) or dead and decaying matter (these are the
saprophytes, e.g.
Saprolegnia). Mixotrophs combine autotrophic and heterotrophic nutrition.

Locomotion for single celled protists is accomplished by means of flagella, cilia, amoeboid and gliding
movements; multicellular protists don’t move. For these single celled organisms moving around in water
is no trivial task. For them, water is an incredibly viscous fluid– flagella and cilia are necessary to propel
them forward. Once they stop beating, because of the viscosity of water, the organisms come to an
abrupt stop as though hitting a wall. Flagella and cilia are therefore crucial mechanisms for locomotion.
Amoeboid movements are much more lethargic and accomplished from the dynamic and complex
cytoskeleton within these organisms.

When it comes to modes of sex and reproduction, we find a great variety of strategies in protists. In this
lab, the modes of sex and reproduction will be covered for only a few selected organisms.

Many books and educators still use the 3 domain (Bacteria, Archaea, Eukarya) and 4 kingdom (Protist,
Fungi, Plants and Animals) system of classification. However, the most recent edition of the Campbell
Biology book (3rd Canadian edition) divides all the divides all eukaryotes into four super groups, namely
Excavata, SAR, Archaeplastida and Unikonta. The kingdoms Fungi, Plants and Animals survived the
5-kingdom system of classification. There are total of 22 groups for protist (highlighted in yellow in the
Figure 1 below). There are two groups of protists on this tree that are not affiliated with any of the four
main supergroups as the evolutionary relationship of these groups of organisms to other eukaryotes is
uncertain. These are haptophytes and cryptomonads.

1
May not be copied or duplicated without express permission of author C. Calogeropoulos
BIOL227 - Lab 3 - Protists F2024

Figure 1: The tree shows phylogenetic hypothesis for the relationships among all the eukaryotes on Erath today.
All eukaryotic groups at the branch tip are related in larger supergroups shown vertically at the end of the tree.
These 4 supergroups are Excavata, SAR, Archaeplastida and Unikonta. Protista are highlighted in yellow.
Source: Campbell Biology. 3rd edition. Page: 630

2
May not be copied or duplicated without express permission of author C. Calogeropoulos
BIOL227 - Lab 3 - Protists F2024

1. Excavates Supergroup
Members of this supergroup have an “excavated” (feeding) groove specialized for phagotrophic
nutrition. They are therefore, mostly, heterotrophs. Its members are also equipped with a single, a pair or
multiple flagella for locomotion. The excavate supergroup includes three clades: the diplomonads,
parabasalids and euglenozoan.

Diplomonads clade
Distinguishing features of clade: Modified mitochondria (called mitosomes) that lack a functioning ETC
making them therefore all anaerobic heterotrophs. Diplomonads also have two to four equal-sized nuclei
and multiple flagella –both features are prominent on stained slides.
Representative included in the lab: Giardia intestinalis

Parabasalids clade
Distinguishing features of clade: Much reduced mitochondria (called hydrogenosomes); its members are
also anaerobic heterotrophs. Most are endosymbionts of animals: in termites, Trichonympha campanula
secretes cellulase to digest the wood ingested by termites. Some parabasalids are parasites: in male and
female reproductive tracts, Trichomonas vaginalis (a sexually transmitted parasite) lives anaerobically
feeding on host cells. T. vaginalis moves around by means of its flagella and undulating membrane.

Representative included in the lab: none

Euglenozoa clade
Distinguishing features of clade: A single of pair of distinct flagella containing a lattice work of proteins
that allow it to whip around and propel the organism forward (recall the viscosity
problem affecting protists’ locomotion). The clade includes the kinetoplastids and euglenids.

• Kinetoplastids have a single large mitochondrion containing a kinetoplast that harbors many
circular DNA molecules. Some are planktonic heterotrophs while others survive as parasites.
Transmitted by blood-feeding insects (African tsetse fly acts as a vector/carrier), Trypanosoma
causes sleeping sickness in humans. In infected humans, the parasite can be seen in blood and
lymph fluids.

Representative included: Trypanosoma

• Euglenids have one or two flagella that emerge from a pocket (the excavated feeding groove
that is unique to the supergroup) at the anterior end of cell. Equipped with chloroplasts,
euglenids can be autotrophic or heterotrophic (hence they are mixotrophs). A specialized
organelle, called the eyespot (or stigma), allows them to detect the wavelengths of light in
their immediate environment. This feature helps them to switch between autotrophic and
heterotrophic nutrition. A proteinaceous strip, called a pellicle, consists of microtubules that
provide strength and flexibility. The pellicle is found outside the cell membrane and allows
them to have slug-like mobility in soil.
Representative included in the lab: Stained slide and live culture of Euglena
3
May not be copied or duplicated without express permission of author C. Calogeropoulos
BIOL227 - Lab 3 - Protists F2024
2. SAR Supergroup
Only DNA similarities unite the diverse members of the SAR supergroup together. This supergroup
includes three major clades: the stramenopiles, alveolates, and rhizarians.

Stramenopile clade
Distinguishing features of clade: A pair of flagella where one is hairy and the other is not. Descended
from an endosymbiotic event involving a red alga, two of the three groups within this clade –the diatoms
and brown algae– are photosynthetic autotrophs. The third member –the oomycetes– lost their plastids
and are therefore heterotrophs.

• Diatoms are unicellular algae (planktonic autotrophs) shaped much like the petri dishes we
find in biology labs. The two halves, called frustules, are porous to facilitated exchanges with
the environment. Frustules are made of silicon dioxide embedded in a matrix of
polysaccharides. The distinct morphology of the frustules helps us to identify diatoms at lower
taxonomic levels (e.g. at the genus and species level). Diatoms are mostly non-motile, but in
some species a mucilage is secreted from the raphe (a narrow opening on the frustules). This
mucilage allows for gliding motion.

Representative included: Navicula

• Brown algae are photoautotrophs, multicellular, and mostly marine.

Representative included: none

• Oomycetes are filamentous protists and include water moulds, white rusts, and downy
mildews. The hyphal nature of oomycetes had them previously classified as fungi. Unlike
fungi, the cell walls of oomycetes are composed of cellulose rather than chitin. Oomycetes
work as important decomposers in aquatic environments, while others are serious plant
parasites causing significant losses in crops each year. Both the decomposers and parasites in
this group derive their nutrition saprophytically.

In addition to similarities in growth form, oomycetes have reproductive structures similar to fungi. The
terminal ends of some hyphae form oogonia, specialized cells that contain eggs. Oogonia are in close
association with antheridia, specialized hyphal cells that produce sperm. When the eggs are fertilized,
they disperse with the aid of water or air currents.
Representative included: Saprolegnia parasitica (a fish parasite)

Figure 2: The figure shows light micrograph of oomycetes and oomycete hyphae on the remnants of a goldfish.

4
May not be copied or duplicated without express permission of author C. Calogeropoulos
BIOL227 - Lab 3 - Protists F2024
The live culture of Saprolegnia shows different types of cells, which are results of either asexual or
sexual reproduction as shown in the figure below.

Figure 3: Life cycle of Saprolengia. Asexual reproduction includes formation of zoosporangia at the tip of
vegetative hyphae. Zoosporangia contained biflagellate primary zoospores which undergoes resting encyst stage
and emerges as secondary zoospore, which undergoes resting phase and eventually form vegetative hyphae.
Sexual reproduction includes structures oogonium and antheridium. Fertilization results in the formation of
oospores and form vegetative hyphae.

Figure source: https://image2.slideserve.com/4197571/slide9-l.jpg

For details on life cycle: https://www.biologydiscussion.com/fungi/life-cycle-of-saprolegnia- with-

diagram-oomycetes/63299

5
May not be copied or duplicated without express permission of author C. Calogeropoulos
BIOL227 - Lab 3 - Protists F2024
Alveolate clade
Distinguishing features of clade: The alveolates represent another clade of the SAR supergroup
descendent from a secondary endosymbiotic event involving red algae. A key feature of its members
includes a pouch-like cavity (or cavities) resembling alveoli (hence its name) just beneath the plasma
membrane. The alveolates include the dinoflagellates, apicomplexans, and ciliates. Of the three, only the
dinoflagellates and some members of the apicomplexans have retained their plastids.

• Dinoflagellates have two flagella located inside of grooves: one transverse, the other
longitudinal. This alignment of flagella gives dinoflagellates the ability to spin as they propel
forward. The cell membrane of these single celled organisms is reinforced with an armor made
of cellulose. Some species of dinoflagellates shed their armor before living as symbionts
inside of animals (e.g. corals). Together with diatoms, dinoflagellates are important
autotrophic plankton.

Representative included in the lab: Ceratium

• Apicomplexans are parasites of animals, and some cause serious human diseases. On one end
of these protist, we find the apical complex of– a collection of specialized organelles that work
to penetrate host cells and tissues. The apicomplexan Plasmodium is the parasite that causes
malaria while Toxoplasma is the cat parasite that causes toxoplasmosis in humans. Both
Plasmodium and Toxoplasma use their apical complex to perforate inside human cells and
tissues.

Representative included in the lab: none

• The ciliates in this supergroup have all lost their red algae symbiont. Consequently, they are
all heterotrophs. Ciliates use cilia to move. The cilia cover the entire surface of the cell. Below
the plasma membrane of ciliates, one can observed the regularly spaced alveoli that are a
distinct feature of the alveolates clade. Ciliates also feature two nuclei: a large macronucleus
and a smaller micronucleus. Sex in ciliates is uncoupled from reproduction. During
conjugation (or sex), two individuals exchange haploid micronuclei while reproduction occurs
as binary fission in a separate event. Contractile vacuoles seen in paramecium expel excess
water in an effort to osmoregulate. Contractile vacuoles appear clear with a star-like pattern
while starch-storing food vacuoles appear grainy and more circular.

Representative included in the lab: Stained slide and live culture of Paramecium

6
May not be copied or duplicated without express permission of author C. Calogeropoulos
BIOL227 - Lab 3 - Protists F2024
Rhizarian clade
Distinguishing features of clade: Rhizarians demarcate themselves by their long thin pseudopods and
mineral shells. The clade includes the forams, cercozoans and radiolarians.

• Forams have porous shells called tests that are reinforced with calcium carbonate. Unlike the
frustules of diatoms, foram tests are single- or multi-chambered. Like diatoms, foram tests in
ocean sediments provide clues about past climates. Furthermore, the coiling pattern of foram
tests gives clues to the surface temperature of our oceans thousands of years ago. Pseudopodia
extend through pores of the test and function both in movement and feeding. Many forams
harbour endosymbiotic algae.

Representative included in the lab: None

• Cercozoans descended from a secondary endosymbiotic event involving a green alga. Most
however are heterotrophs living as parasites. The chlorarachniophytes in this group have
retained their green algal symbiont along with its nucleus. This vestigial nucleus is called a
nucleomorph.

Representative included in the lab: None

• Radiolarians have pseudopodia that radiate out from the central body. Reinforced with
microtubles surrounded with a thin layer of cytoplasm, these pseudopods function to
phagocytize food. Cytoplasmic streaming (powered by the microtubules) brings the food to the
main part of the cell. Like the mineral shells of diatoms and forams, the shells of radiolarians
also accumulate on the ocean floor.

Representative included in the lab: None

3. Archaeplastida Supergroup

Around 2 billion years ago, in a primary endosymbiotic event, a heterotrophic protist engulfed a
cyanobacterium. This event gave rise to the red and green algae clades residing in this supergroup. Also
in the membership of this supergroup are the land plants.

Red algae clade

Red algae are large multicellular protists that harness the shortest wavelengths of visible light (ie blue
and green) that reach deeper in water to photosynthesize. The pigment responsible for absorbing this
light is called phycoerythrin.

Representative included in the lab: None

7
May not be copied or duplicated without express permission of author C. Calogeropoulos
BIOL227 - Lab 3 - Protists F2024
Green algae clade
Their claim to fame is their close affiliation with the kingdom of land plants; green algae and land plants
share the same type of chlorophyl pigments. The clade includes the charophytes and chlorophytes.

• Charophytes share many distinct traits with land plants including the cellulose- synthesizing
protein rings embedded in the plasma membrane of cells and the phragmoplast that helps to
establish a new cell wall after cell division.

Representative included in the lab: Spirogyra.

• The chlorophytes range from unicellular (e.g. Chlamydomonas) to colonial (e.g. Volvox to
filamentous (e.g. Spirogyra). A distinct feature of this group is the pyrenoid bodies within
their chloroplasts which function to reduce rates of photorespiration. Pyrenoid bodies are
responsible for nearly 30% of the global carbon fixed.

Representatives included in the lab: Chlamydomonas and Volvox

4. Unikonts Supergroup

The unikonts supergroup includes two clades: the ameobozoans and opisthokonts. The former clade has
an exclusive protistan membership while the latter includes two protist groups in addition to the fungi
and animal kingdoms.
Ameobozoans clade
Amoebozoans are amoeba that have lobe- or tube-shaped, rather than threadlike, pseudopodia. They
include slime molds, tubulinids, and entamoebas

• Slime molds include the plasmodium and cellular slime molds.

o At one stage of the life cycle of a plasmodium slime mold, the diploid nucleus
contained within a single cell divides mitotically to form a large multinucleated cell.
This mass extends pseudopodia through decomposing material engulfing food by
phagocytosis. Eventually, this mass becomes weblike producing fruiting bodies called
sporangia within which meiosis occurs and haploid spores are discharged. The haploid
spores produce amoeboid or flagellated cells that fuse together to restore the diploid
state.

Representative included in the lab: Physarum

o The cellular slime molds are composed of haploid solitary cells with amoeboid
movements. When food supplies are depleted, these solitary cells aggregate together to
form what appears like a multicellular body. Fruiting bodies (long slender stalks
composed of individual cells stacked upwards) emerge up from the cell aggregate and
spores are released in hopes of finding new sources of food.

Representative included: Dictyostelium discoides

8
May not be copied or duplicated without express permission of author C. Calogeropoulos
BIOL227 - Lab 3 - Protists F2024
Despite lacking a nervous system, slime molds can solve complex mazes and make optimal cost-benefit
foraging decisions. Such an accomplishment is puzzling because solving such complex decision making
is assumed to require a central processing system.

• Tubulinids are amoebas with tube-shaped pseudopodia. Most are heterotrophic consuming
bacteria and other protists. We can find them in ventilation ducts feeding on microbial
biofilms.

Representative included in the lab: Amoeba proteus

• Entamoebas are parasites living within hosts and have a single bulging pseudopod. These
parasites secrete enzymes that degrade human epithelial cells on which they feed. 100,000
people die annually from infections of Entamoeba histolytica.

Representative included in the lab: none

Opisthokonts clade
This clade includes the nucleariids and choanoflagellates protist groups in addition to the fungi and
animal kingdoms. The nucleariids are most like fungi while choanoflagellates are most like animals.
Neither of these protist groups are included in this lab.

Representative included in the lab: none

9
May not be copied or duplicated without express permission of author C. Calogeropoulos
BIOL227 - Lab 3 - Protists F2024
Lab 3: The diversity of protists
Name: Student ID: Tu W Th Bench #:

Important: When drawing the images from the microscope:

-always write the total magnification
-the image should be drawn in the circle and labels should be outside the circle. Make straight arrows
connecting the structure on the image and labelling.

1. Sketch your observation of the stained slide of Giardia sp. Make sure to identify, draw and clearly
label the following features: plasma membrane and nuclei.

Total magnification:

What supergroup does Giardia belong to?

What clade does Giardia belong to?

How does Giardia move?

What disorder is associated with infection of Giardia intestinalis?

How can person get infected by Giardia intestinalis? ___________________________________

10
May not be copied or duplicated without express permission of author C. Calogeropoulos
BIOL227 - Lab 3 - Protists F2024
2. Sketch your observation of the stained slide of Euglena. Make sure to identify, draw and clearly
label the following features: plasma membrane, nucleus, eye spot, and chloroplast.

What other relevant features can you identify from the live culture of Euglena?

Total magnification:

What supergroup does Euglena belong to?

What clade does Euglena belong to?

How does Euglena move?

What is the function of eye spot?

Does Euglena show autotrophy, heterotrophy or mixotrophy and why?

________________________________________________________________________________

11
May not be copied or duplicated without express permission of author C. Calogeropoulos
BIOL227 - Lab 3 - Protists F2024
3. Sketch your observation of the stained slide of Trypanosoma. Make sure to identify, draw and
clearly label the following features: undulating plasma membrane, nucleus, kinetoplast and
flagella.

Total magnification:

What supergroup does Trypanosoma belong to?

What clade does Trypanosoma belong to?

How does Trypanosoma move?

What disease is caused by Trypanosoma?

How is Trypanosoma transmitted? __________________________________________

12
May not be copied or duplicated without express permission of author C. Calogeropoulos
BIOL227 - Lab 3 - Protists F2024

4. Sketch your observation of the stained slide of Navicula. Make sure to identify, draw and clearly
label cell wall and raphe.

Total magnification:

What supergroup does Navicula belong to?

What clade does Navicula belong to?

Why does Navicula shine particularly brightly under dark-field illumination?

13
May not be copied or duplicated without express permission of author C. Calogeropoulos
BIOL227 - Lab 3 - Protists F2024

5. Sketch your observation of the stained slide of Saprolegnia. Make sure to identify, draw and
clearly label the following features: hyphae (mycelium), oogonium, oospores, antheridium, cross
wall, and zoosporangium. For help, use the figure provided in the lab introduction on page 5.

Asexual Sexual

Total magnification:

What supergroup does Saprolegnia belong to?

What clade does Saprolegnia belong to?

Is this protist animal-like, plant-like or fungi like and why? (based on morphology):

14
May not be copied or duplicated without express permission of author C. Calogeropoulos
BIOL227 - Lab 3 - Protists F2024
6. Sketch your observation of the stained slide of Ceratium. Make sure to identify, draw and clearly
label the following features: cell plates, transverse groove and spines (all spines, might not be
visible).

Total magnification:

What supergroup does Ceratium belong to?

What sub-group and clade does Ceratium belong to?

7. Sketch your observation of the stained slide of Paramecium. Make sure to identify, draw and
clearly label the following features: plasma membrane, micronucleus, macronucleus, contractile
vacuole and oral groove.

Total magnification:

What supergroup does Paramecium belong:

What clade does Paramecium belong:

How does Paramecium move:

What is the function of contractile vacuole:

What other relevant feature can you recognize from the live culture of Paramecium? _______________
15
May not be copied or duplicated without express permission of author C. Calogeropoulos
BIOL227 - Lab 3 - Protists F2024
8. Sketch your observation of the stained slide of Chlamydomonas. Make sure to identify, draw and
clearly label the following features: cell wall, chloroplast and nucleus.

What other relevant features can you recognize from the live culture of Chlamydomonas?

Total magnification:

What supergroup does Chlamydomonas belong to?

What clade does Chlamydomonas belong to?

How does Chlamydomonas move to?

What is the function of pyrenoid to?

16
May not be copied or duplicated without express permission of author C. Calogeropoulos
BIOL227 - Lab 3 - Protists F2024
9. Sketch your observation of the stained slide of Volvox. Make sure to identify, draw and clearly label
the following features: mature colony with daughter cells (under low magnification) and mature
daughter cells with vegetative cells and zygotes (under high magnification)
What other relevant features can you recognize from the live culture of Volvox:

Low magnification: High Magnification:

What supergroup does Volvox belong to?

What clade does Volvox belong to?

Is Volvox unicellular, multicellular, colonial ?

Why is Volvox known as green alga ?

Can daughter cells in Volvox live outside of the colonies? Why or why not?

17
May not be copied or duplicated without express permission of author C. Calogeropoulos
BIOL227 - Lab 3 - Protists F2024

10. Sketch your observation of the stained slide of Spirogyra. Make sure to identify, draw and clearly
label the following features: cell wall, chloroplast, nucleus, pyrenoid, single cell in the filament.

Total magnification:

What supergroup does Spirogyra belong to?

What clade does Spirogyra belong to?

What is the shape of chloroplast?

Is this protist animal-like, plant-like or fungi like and why?

18
May not be copied or duplicated without express permission of author C. Calogeropoulos
BIOL227 - Lab 3 - Protists F2024
11. Sketch your observation of the cultured plate of Physarum. Make sure to identify, draw and label
clearly the following: point of origin on plate, food locations (oat flakes) and foraging fan.

What supergroup does Physarum belong to?

What clade does Physarum belong to?

Physarum is a plasmodial slime mold. What are 2 distinct features of plasmodial slime molds.

In contrast to plasmodial slime molds, how will you define cellular slime molds:

___________________________________________________________________________________

19
May not be copied or duplicated without express permission of author C. Calogeropoulos
BIOL227 - Lab 3 - Protists F2024

12. Sketch your observation of the stained slide of Amoeba. Make sure to identify, draw and clearly label
the following features: plasma membrane, pseudopodium, contractile vacuole, food vacuole.
What other relevant features can you discern from the live culture of Amoeba?

Total magnification:

What supergroup does Amoeba belong to?

What clade does Amoeba belong to?

What are pseudopodia?

Is this protist animal-like, plant-like or fungi like and why:

________________________________________________________________________________

13. What was the objective of today’s lab:

_____________________________________________________________________________________________

20
May not be copied or duplicated without express permission of author C. Calogeropoulos
BIOL227 - Lab 3 - Protists F2024

15. From the organisms seen in lab, name 2 protists that are:

Free-living: ___________________________________________________________________

Parasitic: ___________________________________________________________________

Autotrophic: ___________________________________________________________________

Heterotrophic: ___________________________________________________________________

Mixotroph (only 1 protist): _____________________________

16. Circle FALSE options. At the end of the lab, you should:

a. dispose the live culture slides in normal garbage

b. dispose the live culture slides in biohazard waste

c. copy the figures from your lab mates

d. leave the slide/s on the microscope

e. cover the microscope

NOTE: For lab exam, you will be given only pre-stained slides (applicable to all labs).

21
May not be copied or duplicated without express permission of author C. Calogeropoulos