Protista Laboratory

Protists contribute to a group of very diverse organisms that share some common
characteristics. They are unicellular, although they may be found in colonies or in
bodies with specialized structures. As members of the domain Eukarya some are
plant-like and autotrophs (algae), some are heterotrophs (protozoan or animal-like)
and some are more related to fungi and are absorptive heterotrophs. These
organisms exhibit a high degree of complexity. Seventy thousand protozoans have
been identified. While generally microscopic, they are found in damp or wet
environments, such as lakes, oceans, streams, damp soil, mud puddles and
birdbaths.

Objectives:
1. Discuss general characteristics of protists.
2. Discuss general structure of each Phylum.
3. Give examples of each Phylum and explain where members can be found.
4. Compare classification using the 5-kingdom method and 3-domain method.
5. Be familiar with life cycles of Ulva and Chlamydomonas.
6. Differentiate between modes of locomotion in Amoeba, Paramecium and
Euglena.
7. In an evolutionary context, describe the connection between Chlorophyta (green
algae) and higher plants.

Protista

The origin of eukarya is still debated today. Many scientists support the
autogenous theory which suggests that compartmentalization of chemical
processes occurred via segregation of enzymes and subsequent infoldings of
plasma membrane. The serial endogenous theory proposed by Lynn Margulis
suggested that organelles originated via cells engulfing smaller cells with
specialized functions. (E.g.: mitochondria and chloroplasts contain extranuclear
DNA).

Acritarchs are the oldest fossils of protists. Some have been found in Michigan
rocks that are 2.1 billion years old. Protists have been classified quite generally via
nutritional modes as algae (plantlike), protozoa (animal-like) and absorptive
(fungus-like) heterotrophs. In terms of reproduction, all reproduce asexually while
some reproduce sexually via syngamy. Like bacteria, some form cysts when
conditions become harsh.

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Protists are found in wet or damp environments. They inhabit soil, litter, oceans,
ponds and lakes. Some contribute to plankton, the communities of organisms that
drift or swim near the water surface. Phytoplankton constitutes the bases of both
marine and freshwater environments.
Symbiotic relationships exist between some protists and hosts, which may be
classified as mutualistic, commensalistic or parasitic. Hosts may be higher
organisms, such as vertebrates.

The older five-kingdom method of classification is being replaced by a newer

three-domain method of classification. The domains Bacteria and Archaea replace
the kingdom Monera. Reclassification has occurred as more information has been
made available which illuminates the biochemistry and further defines
relationships of representative organisms. Bacteria and Archaea have been
previously presented, thus we will focus on the classification of single-celled
eukaryotic organisms formerly known as protista.

The kingdom Diplomonadida includes organisms that lack mitochondria and have
two nuclei. One example is Giardia lamblia, a diplomonad that causes intestinal
distress and even death. The usual mode of infection is drinking water that
contains human fecal material. Outbreaks of this disease sometimes occur in
daycare centers or other places where good hygiene is in question. Examine
prepared slides of Giardia and note the two nuclei in this flagellated organism.
Draw Giardia.

A newer kingdom, Parabasala includes organisms that lack mitochondria. A

common example is Trichomonas vaginalis the source of a common STD. This
disease can be harbored by males as well as females and is often silent
(asymptomatic) in males. It is an interesting organism that possesses both flagella
as well as an undulating membrane.

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Another candidate kingdom, Euglenazoa, includes the pigmented and flagellated
Euglena as well as another group, the kinetoplastids, a group of parasites that
includes Trypanosoma (found on prepared slides only). Trypanosomes produce
diseases such as sleeping sickness carried by the Tse Tse fly in Africa and Chagas’
disease, in South America. Euglena, generally autotrophic, is typically found in
freshwater environs. They are autotrophs, which occasionally function as
mixotrophs in the absence of light. Examine both prepared slides and living
cultures if available in the laboratory. Living culture media will require a drop of
Detain to slow the organism for viewing. (These slides must be then washed after
viewing with soapy water as the Detain or Protoslo may be very viscous. Detain
puts sets up a carbohydrate matrix which organisms become imprisoned in.) Draw
a diagram of Euglena in the space provided.

Candidate Kingdom Alveolata (a monophyletic group) includes dinoflagellates,

ciliophora and apicomplexa. Dinoflagellates present internal plates of cellulose,
two flagella and are responsible for “Red Tide.” Toxins produced by some of the
dinoflagellates kill organisms in the waters and are responsible for paralytic
seafood poisoning in man. This phylum includes those photosynthesizes that have
external plates of cellulose that exhibit a groove in which one of the flagella can be
found. Examine prepared slides of Cranium and draw a representative.

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Apicomplexa (parasitic Alveolata) have complicated life cycles as in the case of
Plasmodium, which causes malaria. Malaria is responsible for the deaths of 30%
of the children in Africa. The mosquito carries it. The Plasmodia parasite evades
detection by members of the immune system by hiding in cells of the liver. The
genetic code of the parasite has been examined most recently and scientists believe
that a vaccination can be developed against the protist. Draw Plasmodium.

Ciliates (Alveolata) use cilia for motility and feeding. Paramecium, Stentor and
Vorticella are examples of this phylum. Examine prepared slides of paramecium.
Compare the image with that of cells from a living culture. Use a drop of Detain to
slow the motion of the organism. Draw and label a paramecium using your
photographic atlas as a guide.

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The next group is currently unnamed which makes it awkward! We shall call them
the Pseudopods (I did make this one up but common guys, name it!!!) It includes
organisms that use pseudopodia for locomotion, swimming as well as engulfing
other microorganisms (lunch). The Phylum Rhizopoda includes naked and shelled
amoebas with broad pseudopodia. The “false feet” are responsible for locomotion
as well as engulfing prey. Amoeba move very slowly and detain is not
recommended for use in slide preparation. Amoebas are usually colorless and it is
often necessary to change both the intensity and angle of the light to observe them
successfully. (Light rheostat and iris diaphragm) Draw an amoeba in the space
provided.

Members of Actinopoda (Pseudopodia) are found in planktonic habitats. They

tend to be spherically symmetrical with axopodia (heliozoans are fresh water
organisms while radiozoans are marine). Examine prepared slides of radiolarians
and draw example of the organism. Fresh cultures of Actinopoda will display sun-
like structures with axopodial rays. Draw examples of heliozoans and radiolarians.

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Foraminifera (Pseudopodia) are protists that have calcareous compartments with
pseudopodia. They are structurally responsible for the creation of the White Cliffs
of Dover in England. Obtain a slide of forams and draw representatives. Note the
various compartments. Examination of forams as well as other representatives has
given geologists much information on the formation of landmasses.

Mycetozoa include Myxogastrida, acellular slime molds (coenocytic plasmodium).

These include organisms that are fungus-like heterotrophs and are often brightly
pigmented. The feeding stage is a mass while Dictyostelida’s (cellular slime mold)
feeding stage is a single haploid cell. The slime molds are probably most related to
amoeboid protists. Note the illustrations in the photographic atlas.

The Candidate Kingdom Stramenopila, another monophyletic group, includes

diatoms possessing silica shells that overlap. The Bacillariophyta produce oil,
which promotes buoyancy, and contain a number of pigments. They are used in
production of filters and in polishing agents. Obtain a prepared slide of diatoms
and draw representatives. These organisms, along with many others have been
used in forensic science to establish locations as the kinds and numbers tend to be
fingerprints of aquatic environs.

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Stramenopila also includes golden algae, Chrysophyta, which are biflagellates,
freshwater and colonial. Examples may not be available in the laboratory. Brown
algae, Phaeophyta, is also a member of this candidate kingdom. Phaeophyta, are
the largest algae. The pigment fucoxanthin produces the brown color. Examples
are seaweeds such as Fucus and Sargassum. The basic structure of the algal body
is holdfast (root-like structure), stipe (stem) and blade (leaf-like structure.)
Extracts of algin, a polysaccharide in the cell wall, are used as thickening agents in
food preparation. Examine the examples of brown algae available in the
laboratory. Sketch representatives.

Rhodophyta, another candidate kingdom, includes red algae, which survive in

deeper water than other algae due to the presence of the pigments phycocyanin and
phycoerythrin. These pigments absorb EMR in the green and blue wavelengths.
The substance, agar, used in the culture of microbes is obtained from the wall of
the red algae. Another extract, carageenan, is used to thicken ice creams, puddings
and soups. Red algae are used as wrapping for Sushi. Porphyra is available in the
classroom for observation.

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The green algae, Chlorophyta, are not clearly classified as Rhodophyta or Plantae.
Green algae are generally found in most fresh-water environs. They include
unicellular (motile and non-motile) as well as colonial and filamentous forms.
Some members form symbiotic relationships in lichens and are thought to be the
ancestors of green plants due to similar biochemistry. Examples of Chlorophyta
include: Ulva, Spirogyra, Chlamydomonas, Chara, Volvox. Examine the prepared
slides as well as fresh cultures of the above green algae. Draw representatives.

Seaweed includes alga from Phaeophyta, Rhodophyta, and Chlorophyta.

Specialized structures exist: the thallus (body), holdfast (rootlike) and blade
(leaflike.) Cell walls of cellulose and gel-forming polysaccharides sometimes
with calcium carbonate make them unpalatable but sturdy for intertidal zones.
Uses include thickener in foods (algin, agar and carageenan), lubricants, and in
microbiological culture medium.

Recent revelations concerning the biochemistry of these protists have given cause
for reclassification. Many similarities exist between green algae and plants. These
will be discussed later. While frustration exists concerning taxonomy in this
incredibly diverse group, more information becomes available daily, allowing us
the opportunity to reexamine phylogenetic relationships.

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Laboratory Questions
1. The ancestors of protists were the first to have organelles and structures such as:

2. Where are protists found?

3. List protists that photosynthesize.

4. List protists that resemble absorptive heterotrophs.

5. List protists that are nourished via ingestion.

6. Give examples of and distinguishing characteristics of:

a. Euglenoids:
Euglenophyta-

Kinetoplastids-

b. Alveolata:
Dinoflaggelata-

Apicomplexa-

Ciliophora-

7. Stramenopilia:
Bacillariophyta (diatoms)-

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 Chrysophyta (golden algae)-

Phaeophyta-

8. Rhodophyta (red algae)-

9. Chlorophyta (green algae)-

Classification of these related protists is related but unclear.

10. Rhizopoda-

11. Foraminifera-

12. Actinopoda-

13. Describe the general structure of a sea-weed.

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