Kingdom Protista

• Sometimes called the “Junk drawer”

• Contains animal-like, plant-like and

fungus-like organisms

• Plankton (zoo-/phyto-/myco)
 CHARACTERISTICS OF PROTOZOANS
• Unicellular
• Eukaryotic
• Basically Ingestive Heterotrophs
• Lack cell walls but have definite shapes
• Most are motile
• Basically reproduce by asexual reproduction
• Aerobic but some can live in anaerobic
conditions (ones living in digestive tracts)
 Special structures:

1. Macronucleus – controls metabolism

2. Micronucleus - involved in conjugation
3. Contractile vacuoles – maintains
4. Meostasis
5. Ingestion structures
6. Anal pore – excretion of wastes
7. Trichocysts – defense mechanism
 HABITAT

• Majority of free-living
• Marine, terrestrial & freshwater.
• Some are parasites on algae to vertebrates
• Make up the zooplankton in marine
ecosystems. Feed on phytoplankton
• Abundant in soil or on plants & animals
• Some live in guts of termites, roaches &
ruminants (cows)
 DIFFERENT PROTOZOANS

Paramecium with trichocysts

Paramecium

Didinium

Vorticella
Stenor
 PARAMECIUM

• Paramecium is a
small unicellular
organism.
• It is plentiful in
freshwater ponds.
STRUCTURE
Position Of Protists In The
Prokaryotic Kingdom
 Classification
Classified by method of locomotion
• Mastigophora – have one or more flagella
- Have a flagella with a 9-2 microtubule
arrangement
- Flagella are polar & undulates, pushing
protozoan in opposite direction
- Longitudinal reproduction
i.e. Peranema,
Chilomonas
• Ciliata (Ciliophora)
- Have cilia.
- Similar in structure to flagella but shorter and all
over surface of organisms
- Cilia usually arranged in rows & connected to each
other
- Cilia near oral cavity involved w/ food getting
- Transverse fission, & sexual repro by conjugation
Ie – Paramecium, Didinium, Blepharisma,
Vorticella, Stentor
• Sarcodina
- Use Pseudopods for movement
- Cytoplasmic streaming – amoeboid
movement
- Tips of pseudopods are less viscous so
flow goes in that direction
- Pseudopods for phagocytosis
- Reproduce by binary fission
i.e. Amoeba, Naegleria, Heliozoans, Radiolarians,
Foraminifera
• Sporozoa
– No method of motility
– All are parasites – use host for motility
– Reproduce by schizogamy (multiple fission) in
host & sexual reproduction in a second host
Ie. Plasmodium (malaria), Giardia, Toxoplasma,
Trypanosoma, Trichomonas
Animal-Like Protists
PROTOZOAN PROTIST
 EVOLUTION
•Evolved from the Archae approx. 1.5 billion years ago

•Polyphyletic group- protists arose by way of more than

one ancestral group

•Represents separate evolutionary lineages

•Plant like b/c autotrophic (produce their own food)

•Animal-Like b/c they are heterotrophic (feed upon other

organisms)
Figure 8.20
TYPES OF PROTOZOANS

• Zooflagellated Protozoans

• Flagellated Protozoans

• Phytoflagellated Protozoans
 Zooflagellated Protozoa
• Lack chloroplast
• Heterotrophic
• Some members are important human
parasites

• Species Trypanosoma brucei cause African

sleeping sickness (Intermediate host- Tsetse
flies )
Figure 8.8 (b)
 Flagellated Protozoa
• Flagellates are the ancestors of ameoboid
protozoan

• Phytoflagellated (photosynthesizing)

• Zooflagellated (particle feeding and

parasitic)
 pellicle

contractile
macronucleus
vacuole

cytoplasm:
ectoplasm
micronucleus
endoplasm

cilia

oral groove

anal pore gullet

food
vacuole
 Phytoflagellated Protozoa
• Chlorophyll (oxygen for marine life)

• One or two flagella

• These protozoans are large portion of the

marine food i.e dinoflagellates

• Two flagellates, chlorophyll, xanthophyll

(bloom=red tides) and results in fill kills (Red
sea, bible)
Figure 8.6
 Other Phytoflagellated Protozoa
Euglena
• Freshwater
phytoflagellated protozoa
• Chloroplast has a pyrenoid
(synthesizes and stores carbohydrates)
• Feed by absorption or are
heterotrophic
• Stigma- photoreceptor at the base of
the flagellum
• Haploid organisms and reproduce
binary fission
 FUNGUS LIKE PROTISTS

• Like fungi, they are heterotrophs, have cell walls, use spores to reproduce.
Unlike fungi, they can move at some points in their life cycle.

Three types:
• - Water Molds and Downy Mildews:
• Both types live in water or moist places, & look like
fuzzy threads.
• They attack food crops (potatoes)
• - Slime Moulds:
Live in moist soil and on decaying plants and trees.
Some are with beautiful colors. They move using
• pseudopods.
• They eat bacteria and other microorganisms.
Can combine, forming a multicellular mass & spores.
Spores develop into a new generations of slime moulds
 PLANT LIKE PROTISTS

Called algae & are autotrophs (pigments & photosynthesis).

* Some are unicellular, living unconnected from other algae cells.

* Others form colonies together with a few cells specializing for

reproduction, etc. (Most colony cells continue to carry out all
normal functions.)

* Some algae are multicellular like seaweed, where all cells are
specialized.
Paramecium Movement
• The outer surface of the cell is covered
with many hundreds of tiny hair-like
structures called cilia.
• These act like microscopic oars to push
through the water, enabling the
organism to swim.
• If Paramecium comes across an
obstacle, it stops, reverses the beating
of the cilia, swims backwards, turns
through an angle and moves forward
again on a slightly different course.
• It moves so quickly that we have to add
a thickening agent or quieting solution
to the slide to slow it down to study it.
 Paramecium Feeding
• Paramecium has a permanent feeding
mechanism, consisting of an oral groove and a
funnel-shaped gullet into which food is drawn by
the combined action of cilia which cover the
body and other cilia lining the oral groove and
the gullet.
• As it moves through the water it rotates on its
axis and small particles of debris and food are
collected and swept into the gullet.
• They feed on small organisms such as bacteria,
yeasts, algae and even other smaller protozoa.
Paramecium Excretion
• Food waste left in a food
vacuole is excreted through
the anal pore (the vacuole
and pore fuse.

• Other wastes left over from

cellular activity (metabolic
waste) simply diffuse through
the pellicle.

• Excess water and some

metabolic wastes are
excreted through the
contractile vacuole.
ASEXUAL REPRODUCTION IN
PROTOZOANS
Asexual Reproduction in Protozoa
Conjugation in Paramecium
Paramecium Reproduction
• In favourable conditions the
cell divides in two by a process
called binary fission (asexual
reproduction).
• This forms two new cells, each
of which rapidly grows any
new structures required and
increases in size.
• This whole process may take
place two or three times a day
if conditions were right.
 Paramecium Reproduction
• This is a more complicated
method called conjugation
(sexual reproduction).
• It involves two cells coming
together to exchange nuclear
material.
• The two cells then separate
and continue to reproduce by
simple division.
• It is similar in some ways to
sexual reproduction in more
complex animals.
Reproduction in Ciliates
Paramecium
conjugating

Transverse Binary
fission
 Symbiotic lifestyles

• Symbiosis

• Parasitism- a form of symbiosis- organism

lives in or on other (Host)
 Other kinds of symbiosis
• Don’t harm host

– Commensalisms- one member benefits

– Mutualism- both benefit

 Some parasites have life cycles involving
multiple hosts

• Definite host- harbors the sexual stages of

the parasite

• Intermediate host- the offspring enter

another host where they reproduce asexually,
to complete lifecycle the final asexual stage
must have access to a Definite host
 Paramecium Grammers
• Growth: 0.05 mm avg.
• Respond:
-react to chemicals – salt and vinegar
-live in slightly acidic environments (stagnant
H2O)
-anterior end sensitive – move by trial and
error
 Paramecium continued…

• Adaptations:
– Cilia to help feed and escape
– Contractile vacuoles
– Trichocysts
 Paramecium continued…
• Movement: by cilia in circular motion, move ~ 60
mm/hr

• Metabolism:
-food pulled into oral groove by cilia
-food vacuole forms at gullet
-lysosome aids with digestion

• Feed mostly on bacteria, smaller protozoans and

algae.
 Paramecium continued…
• Excretion:
-Contractile vacuole removes excess H20.
-C02 across pellicle by diffusion
-anal pore removes waste.
• Reproduction: Asexual - cell division
Sexual – conjugation (exchange
of micronucleus DNA)
CLADOGRAM OF PROTOZOA
RELATIONSHIPS
.
Endosymbiosis and Cytoplasmic
Inheritance in Paramecium
This Topic Will Focus On The Following :

Altenburg paper (1948) Current understanding

–Plasmagene hypothesis of Kappa bodies (Preer
–Kappa body symbiosis 1974)

Other cytoplasmic inheritance Paramecium biology

in Paramecium (Meyer 2002) –Cell biology
–Life cycle
Altenburg paper (1948) investigates the evidence that
Kappa bodies are a symbiont

Kappa bodies are elements within Paramecium that cause

them to be killers

Killer Paramecium kill other Paramecium in the

immediate environment

Kappa particles, thought to be plasmagenes by Sonneborn, but Altenburg

suggest they may be symbionts
The plasmagene theory suggested kappa bodies were
genes within the cytoplasm
Plasmagenes defined as self-replicating structure capable of
producing traits that exist in the cytoplasm and are independent of
chromosomal genes.

The trait that Kappa bodies produce is the killing factor

Kappa bodies are inherited through the cytoplasm and not through
chromosomes

Sonneborn wrote in 1976, “It was awful of me to be so attached to a pet

idea. That was an ordeal between my mind and my heart and it took a
while for the mind to win and the heart to accept. Impersonal scientific
objectivity is a goal to be sought by hard self-discipline; we are not born
with it.”
Altenburg’s evidence that Kappa bodies are symbionts is
strongly supported by evidence

Preer (1948) showed Kappa is large enough to see under a

light microscope

38o C kills Kappa but not Paramecium

Division of Kappa and Paramecium is independent of each

other
Paramecium with symbiont
There is an upper limit of the number of Kappa in Paramecium

More likely a symbiont than a parasite

Preer (1974) reviewed the overwhelming evidence that
Kappa bodies are symbionts

Kappa contains DNA, RNA, protein, and lipids in

proportions expected in bacteria

Kappa contains electron transport system with

cytochromes similar to bacteria and not eukaryotes
Electron micrograph of symbionts

Electron microscopy clearly showed that Kappa is

prokaryotic

Electron micrograph of flagellated Kappa

Current information has shown why Kappa induces killing
and the different types of bacteria symbiosis

Kappa bodies kill other Paramecium by releasing toxins into the

environment

The presence of the symbiont makes the host resistant to the toxin

Kappa bodies are transmitted by the cytoplasm during asexual division

Many other types of symbionts found

sigma gamma

Kappa is the most common lambda alpha

pi delta
omega
mu
The discovery of bacterial symbionts within Paramecium
allows for their taxonomic classification

Kappa, mu, gamma, and nu are in genera Caedobacter

Alpha bodies are in the genera Cytophaga

Lambda and sigma are in genera Lyticum

Delta bodies are in genera Tectobacter

Differences have been found between Kappa bodies in the
same host

Some Kappa bodies contain refractile ( R ) bodies

R body is a type of inclusion body

When genes from one organism are within

another organism and are transcribed, a
inactive protein may form

Magnified image of coiled R body (2)

Kappa bodies may contain ‘R’ bodies and it affects their
reproductive capability

Non bright Kappa bodies do not contain R bodies but can

reproduce

Bright Kappa bodies do contain R bodies but cannot

reproduce

Dividing symbiont

Non bright produce other non bright, but occasionally a non

bright turns into a bright

Toxicity associated only with Brights

There is still unsolved questions regarding Kappa body
symbiosis

What benefit does Paramecium get from the

symbiosis?

How does the presence of a Kappa body induce resistance to the

toxin?

Resistance can be overcome with large toxin dose

The presence of Kappa with or without R bodies

induces resistance to the toxin
Other types of cytoplasmic inheritance discovered in Paramecium and
other ciliates is:

Genome-wide DNA rearrangements

Mating type

Serotypes
Paramecium has a complex cellular biology

Eukaryotic

Ciliates contain at least 2 nuclei

Germ-line micronucleus (MIC)
Somatic macronucleus (MAC)

MAC is generated from the MIC

Diagram of Paramecium
Extensive genome rearrangements occur in the MAC
The two nuclei make the life cycle of Paramecium more
complicated than other eukaryotes
MIC goes through meiosis and the haploid
MIC goes through mitosis
Result is 4 haploid MIC, but 2 are
degraded

Paramecium exchange 1 haploid MIC

MIC fuse and form diploid MIC and

duplicate via mitosis

Old MAC degrades and duplicated MIC

is processed into new MAC

In asexual reproduction, the MIC goes

through mitosis and the MAC goes through
amitosis
Genome-wide rearrangements of the MAC genome
consists of deletion of DNA sequences and chromosome
amplification
The developing new MAC loses 10 - 95% of
the genome depending on the ciliate

MAC chromosomes are amplified to a high

ploidy level

Deletion occurs after an initial amplification

of the MIC genome but before the ploidy
level is reached
The deletion of DNA is located at specific sequences called
internal excised sequences (IES)

IES are located in coding and non-coding regions of the MIC genome

These sequences are not present in the MAC genome

At some point in MAC development, the IES sequences are deleted

The mating type of Paramecium
shows maternal inheritance

Conjugation of P. caudatum by Yanagi

Paramecium has 2 mating types - O and E

Both are not determined by genetic differences as they are both produced
in homozygous wild-type strains

Mating type is the same through asexual reproduction but can change after
sexual conjugation and MAC formation

After conjugation O cells mostly produce other O cells and E cells produce
other E cells
Paramecium mating types do not follow the
Mendelian segregation of alleles

A. Mendelian segregation of allelic pairs

B. Maternal inheritance of mating types
Mating types O and E depends on different states
of MAC genome

Transferring E maternal MAC into O cell causes the progeny to become E

Transferring O MAC does not change E cells

O is the default mating type

E cell O cell E cell

Produces

Insert E MAC
This differential state of MAC is dependent on the
presence of IES in the MAC

The mutation mTFE causes O cells to become E

This mutation affects the excision of an IES on the G gene

The G gene is a surface antigen and the failure of

excision causes a nonfunctional protein to be
translated

excision
Functional - type O

Mutational Nonfunctional - type E

MIC G gene retention

MAC G gene
Microinjection studies have shown that the presence of
an IES sequence in the MAC inhibits the excision of its
homologous IES in the MIC

O cells contain G gene in the MAC without its IES (IES-)

E cells contain the G gene in the MAC with its IES (IES+)

Injecting a plasmid of IES+ G gene into O cell’s MAC created the retention of the
IES in the MAC of daughter cells

Injection of IES- plasmid did not induce excision

The presence of IES in the MAC causes the retention of the IES in subsequent
generations after sexual conjugation
Microinjection of IES+ plasmid retains the IES in the
MAC genome after autogamy
Meyer (2002) asked, “How can a sequence introduced in one
nucleus affect the excision of the homologous sequence in another
nucleus?”

Two models developed

Model 1: Sequence-specific protein factors are required for

the excision of the IES in the developing MAC

The problem with this model is the large

number of protein factors needed, about
50,000

Model 2: Sequence specificity is achieved by homologous nucleic acid (most likely

RNA) that is transported from the maternal MAC to the developing MAC
Mochizuki (2004) explained the Scanning Model, a synthesis
of Meyer’s model 1 and 2

Entire MIC genome is transcribed bi-

directionally and forms dsRNA

dsRNA is cut up into smaller RNA

called scn RNA

Scn RNA move to the old MAC and any

matching homologous sequences are
degraded

Scn RNA that were not degraded move to

the developing MAC

These scn RNAs target homologous

sequences which are deleted in an RNA
i-like mechanism
 Polymorphic lifestyles

• Have different forms during their life cycle

• May form cysts (vegetative cells) when
adverse conditions exist. Cysts are not heat
and chemically resistant.
 SUMMARY

Paramecium has many instances of cytoplasmic and maternal

inheritance

Paramecium
Kappa bodies are bacterial symbionts that produce a killing
factor and they are inherited through the cytoplasm

IES excision and retention in the MAC is maternally inherited by

the genome present in the MAC
Electron micrograph of Kappa
 Ecological importance
• Members of the food chain – Primary or
Secondary consumers
• Consume soil bacteria & algae (1 paramecium
can ingest 5 million bacteria/day
• Involved in sewage disposal by metabolizing
nutrients present to carbon dioxide & water
 HARMS
• Cause disease in host organisms
Malaria – Plasmodium via mosquito
Toxoplasmosis – Toxoplasma
African Sleeping Sickness – Trypanosoma
via tsetse fly
Chagas – Toxoplasma
Vaginitis – Trichomonas
Giardiasis – Giardia
150 million people/year in world contract Malaria &
1.5 mill/year die of it.
FACTS ABOUT PARAMECIUM
“One scientist calculated that if all the
progeny of a single Paramecium
survived, assuming a division rate of
once a day, then after 113 days, the
mass of paramecia would equal the
volume of the Earth! “