The Origin and Evolution of Microbial Life: Prokaryotes and Protists

I. Origin of life on Earth

A. The following represents a superficial overview of science's best explanation for how
life on Earth may have formed from nonliving materials.
1. Much of this topic is speculative since no one was around to record the events
2. Vast majority of the ideas and theories are based on small amount of evidence
3. However, the generalized theories presented are accepted by most of the scientific
community today.
4. When Earth was formed, the atmosphere lacked free oxygen.
5. Water in the form of liquid accumulated as the temperature cooled. Due to its size
and distance from the sun, the earth could retain water in the liquid state.
6. Rains eroded away many minerals from rocks.
7. Various sources of energy on early earth that caused chemical reactions to take
place: volcanic activity, lightening storms, high levels of radiation that bombarded
the earth's surface since the atmosphere was not well developed.
8. No one knows exactly when, where, or how life originated - no fossil records exist
from that far back in time due to the tremendous amount of geological activity that
has occurred since then.
B. Nonliving materials (simple molecules) combined into more complex molecules that
were eventually able to reproduce themselves.
1. The chemical evolution of life, which suggests that complex organic molecules
(necessary components of life) originated from nonliving simple molecules.
2. The original idea was proposed by a Russian biochemist Alexander I. Oparin
(1922).
3. Organic molecules formed from atmospheric gases.
a. These organic molecules collected in a thin soup in shallow seas and lakes.
b. As these waters evaporated, those organic molecules became concentrated.
c. Radiation, lightening, volcanic activity, etc. provided the energy necessary to
allow reactions to occur - from this Oparin suggested that life could arise.
4. However, Oparin lacked the technology to test this hypothesis
5. Amino acids, purines, and pyrimidines formed from simpler molecules (those
available in the reducing atmosphere)
a. Stanley Miller's experiments - S. Miller & H. Urey (1953) read of Oparin's
ideas and set out to prove that organic molecules including amino acids and
purines and pyrimidines could be produced in these conditions.
(1) They set up a small environmental chamber and duplicated conditions of
the early atmosphere described by Oparin
(2) Used a mixture of hydrogen, methane, ammonia, and water vapor
(3) A spark of electricity served as an energy source (mimicked lightening)
(4) After allowing this experiment to run for only 1 week, simple organic
compounds such as hydrogen cyanide, formaldehyde, cyanogen,
acetaldehyde, cyanoacetylene, and proprionaldehyde were formed.

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 (5) These simple compounds dissolved in water and were rapidly converted
into amino acids, simple acids, purines, and pyrimidines- the building
blocks of life.
6. Monomers combined to form polymers - The next stage in the sequence leading to
life was the generation of large molecules by polymerization of small molecules.
a. The important large molecules which organism are composed of include
polysaccharides, proteins, and nucleic acids.
b. These are all formed by the combination of the subunits or monomers. The
reactions leading to these large molecules are condensation or dehydration
reactions, which produces water in the process. .
7. Nucleic acid polymers can copy themselves (chemical reproduction)
a. Some polymers can essentially copy themselves.
b. Nucleic acids, the basis of today's genetic code, are clearly capable of
self-copying, and the purine and pyrimidine parts of nucleotides were formed
under the conditions simulated in Miller's experiment.
c. Others have now done similar experiments, forming many different organic
molecules, including amino acids, lipids, sugars, and nucleotides.
8. Very simple life probably evolved randomly in this manner over about 300 million
years.
9. First living cells were probably only lipid membrane-bound sacs containing simple
nucleic acids that manufactured proteins.
C. Main path of information in living organisms is DNA -> RNA -> protein
1. Proteins are synthesized by a process that begins with transcription of information
from DNA to an RNA molecule that has a base sequence complementary to that of
one strand of the DNA.
a. The mRNA is then used to synthesize a specific polypeptide from amino acids.
b. Amino acids are brought to a ribosome by specific tRNA molecules and are
attached sequentially to the growing polypeptide.
2. How could such a system evolve if proteins catalysts needed nucleic acids for their
formation but nucleic acids needed proteins to catalyze their own replication?
Which came first?
D. Problem - proteins (enzymes) are used in all steps of this process
1. DNA synthesis (replication)
2. Protein synthesis
3. But proteins are constructed according to info in nucleic acids
E. So how did nucleic acids copy themselves without enzymes for copying themselves?
1. The inability to solve this problem held up research on the origin of life for several
decades.
2. Discovery of ribozymes (catalytic) - RNA molecules that have catalytic ability
3. Hypothesis - RNA was the first information carrying molecule and some RNA had
catalytic capability
a. Biochemists believe that the first information-carrying molecules were short
strands of RNA that replicated themselves without the help of enzymes.

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 b. Evidence that RNAs can replicate themselves came first from experiments
conducted by Manfred Eigen in the late 1970's. Eigen added RNA molecules
to solutions containing monomers for making more RNA and found that
sequences of 5 to 10 nucleotides formed.
c. Although these experiments showed that RNA could replicate itself, they did
not demonstrate that RNA could catalyze the synthesis of other molecules as
true enzymes do.
d. Within two years, however, studies of a tRNA-processing enzyme showed that
RNA could catalyze reaction without the protein enzymes.
F. How did first cells form?
1. Many different solutions that contain polymers will form coacervates (round
structures similar to a cell membrane) when shaken.
a. Coacervates are quite stable and will form in solutions of many different types
of polymers.
b. When added to a solution of coacervates, many substances are concentrated
within the drops. For example, lipids will coat the drops with membrane like
structures, which strengthens the drops and helps control the rates of passage of
materials in and out.
c. In early conditions on earth, the concentrated RNA molecules combined with
lipids would also display the property of surrounding droplets of polymers
acting as a membrane. These RNA molecules which are capable of
self-replication would tend to produce other membranes.
2. Membranes would have made homeostasis possible. - The key advantage of having
membrane bound systems is that it makes homeostasis possible.
II. Earliest prokaryote fossils - ca. 3.5 billion years old (3.8 billion years ago - estimated time of
life's origin)
III. Early bacteria that evolved the ability to use water as a source of H for their metabolism
liberated O2
A. The oxygen concentration increased because certain sulfur bacteria evolved the ability
to use water as the source of hydrogen during photosynthesis.
B. The cyanobacteria that evolved from these sulfur bacteria became very abundant. They
liberated enough oxygen to open the way for the evolution of oxidation reactions as the
energy source for the synthesis of ATP
C. O2 levels in atmosphere have been gradually increasing during Earth history.
IV. Only tiny cells would be able to survive in the low-oxygen environment of early Earth
(because of the cellular surface area/volume ratio)
A. Small unicellular organisms can obtain enough oxygen by simple diffusion even when
oxygen concentrations are very low.
B. Larger unicellular organisms have a lower surface area to volume ratio. In order for the
larger celled organisms to get enough oxygen by simple diffusion, they must live in an
environment where concentrations of oxygen are higher.
C. Eventually enough O2 was available to support multicellular life

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-------------------------------------------- End of Exam 1 Material--------------------------------------------

V. Diversity of life
A. Approximately 1.5 million known species
B. Probably 10-50 million species living on planet
C. Most species that have ever lived are extinct
D. Current extinction rates are estimated to be very high (on the order of a “mass
extinction”)
VI. Prokaryotes
A. Domain Bacteria has one kingdom Eubacteria “true bacteria”
B. Domain Archaea has one kingdom Archaebacteria, “ancient bacteria”
C. The two domains that make up prokaryotes were thought to be very closely related
because of their similar appearance and were placed in a single Kingdom Monera.
D. Not until the 1970's did we realize how different they are. In 1996, the genomes of
Archaea were sequenced and we found that they are more different than we expected.
E. Very different metabolic pathways
VII. General characteristics of prokaryotes
A. Tiny - can’t be seen with naked eye
B. Numerous - more individuals than any other group (ca. 4800 species known)
1. Bacteria in one person's intestinal tract outnumber all the humans who have ever
lived.
2. Although they are small, they perform many critical roles in the biosphere
3. All are unicellular, but may be grouped in chains or filaments
C. Occur in many habitats, examples:
1. Prokaryotes are much more diverse than any other group with respect to the habitats
they occupy.
2. They have spread to every kind of habitat from the coldest to the hottest, from the
most acidic to the most alkaline, and to the saltiest.
a. Bottom of ocean
b. In rock 2 km below surface of earth
c. Inside other organisms
d. Hot springs
e. Ice
D. Lack a true nucleus (nucleoid, instead)
E. Lack a cytoskeleton
F. Lack membrane-bound organelles (but may have complex internal membranes)
G. No mitosis, no meiosis
H. There are three shapes that are particularly common among the prokaryotes.
1. Spheres - coccus (cocci), can live singly or may form chains, plates and blocks of
cells
2. Rods - bacillus (bacilli) singles or chain
3. Spiral - spirillum, single or chain

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I. Most have a cell wall
1. Most prokaryotes have a thick cell wall, which is different from the cell wall in
plants and algae that contains cellulose and other polysaccharides
2. Almost all bacteria have cells walls containing peptidoglycan (a polymer of amino
sugars).
3. Archaeans cell walls are of differing types, but most contain significant amounts of
protein (often a glycoprotein).
4. Peptidoglycan is unique to bacteria, its absence from Archaea is one of the key
differences between the two.
5. In 1884, Hans Christian Gram, a Danish physician, developed a simple staining
process that has lasted into the modern era.
6. The technique is to stain a smear of cells with a violet dye; it is then washed with
alcohol and counterstained with safranine (a red dye).
a. Gram positive - gram staining technique stains them purple; large amount of
peptidoglycan
b. Gram negative - gram staining technique stains them pink to red; little to no
peptidoglycan
J. Locomotion
1. Many are non-motile
2. Rolling, gliding
a. Spirochetes use a rolling motion made possible by internal fibrils
b. Many cyanobacteria use a gliding mechanism that is poorly understood.
3. Flagella (made of flagellin) - The most common type of motion is driven by
flagella, whiplike filaments that extend singly or in groups from one or both ends of
the cell, or all around it.
K. Reproduction
1. Asexual - through fission or spores
2. Some genetic recombination occurs (e.g., through conjugation) - Conjugation
occurs when to bacteria line up side-by-side and exchange some DNA material
through a small tube
3. Mutation is primary source of genetic variation
4. Generation time can be very rapid (e.g., E. coli can double its population size every
20 minutes) The shortest known prokaryote generation time is about 10 minutes,
but values of 1 to 3 hours are common
5. Generation time can be very slow (e.g., some can remain “dormant” for long
periods of time in between divisions); Bacteria living in rock deep in Earth's crust
may suspend growth for more than a century without dividing and then grow for a
few days before suspending growth again.
L. Metabolism - In the long evolutionary history of prokaryotes, in the extremely diverse
environments where they exist, there are many forms of metabolism that have evolved.
1. O2 use
a. Obligate anaerobes - (die if exposed to O2) Prokaryotes that can live only by
anaerobic metabolism (fermentation), they are poisoned by gas.

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 b. Facultative anaerobes - (can survive in presence or absence of O2) Anaerobic
and aerobic modes (fermentation and cellular respiration)
c. Obligate aerobes - (die without O2) Others are unable to survive for extended
periods without oxygen.
2. Nutritional categories
a. Photoautotrophs - use light as energy source, CO2 as C source (some Bacteria,
some protists, almost all plants)
(1) e.g. cyanobacteria uses chlorophyll a and releases oxygen as a byproduct
(2) Some use bacteriochlorophyll and don't release oxygen, but release sulfur
instead.
b. Photoheterotrophs - use light as energy source, organic molecules produced by
other organisms as C source (some Bacteria)
(1) Use compounds such as carbohydrates, fatty acids, and alcohols as their
organic “food”
(2) e.g. Purple nonsulfur bacteria
c. Chemoautotrophs - use inorganic substances as energy source, CO2 as C source
(some Bacteria, some Archaea)
(1) Similar to the carbon reduction cycle in photosynthesis
(2) In 1977, scientists exploring the Galapagos Islands discovered a
spectacular example of chemoautotrophy. They found an entire ecosystem
based on these bacteria that are incorporated into a large community of
crabs, mollusks, and giant worms. At a depth of 2500 m, below any hint
of light from the sun.
d. Chemoheterotrophs - use organic molecules produced by other organisms as
energy and C source (most Bacteria, Archaea, many protists, all animals, fungi)
VIII. Benefits to other organisms
A. The nitrogen cycle depends on a number of species of Bacteria.
1. All organisms need nitrogen for their proteins and nucleic acids.
2. Without nitrogen fixing and denitrifying bacteria, nitrogen would be unavailable to
organisms.
3. Nitrogen fixers - bacteria that convert N2 into molecules useable by them and other
organisms (NH3)
4. Denitrifiers - bacteria that return nitrogen to the atmosphere as N2
B. Mutualism and commensalism
1. Endosymbiosis - mitochondria and chloroplasts are thought to be the descendants
of free-living prokaryotes
2. Root nodules in some plants - Some plants formed an association with bacteria of
the genus Rhizobium to form cooperative nitrogen-fixing nodules on their roots.
3. Digestive tracts of many animals
a. Many animals, including humans, have a variety of bacteria and archea in their
digestive tracts.
b. Cows depend on prokaryotes in their complicated stomachs to perform
important steps in digestion.

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 c. Cows cannot produce cellulase, the enzyme needed to start digestion of
cellulose, but some of their stomach bacteria produce cellulase
4. Many bacteria live in and on human bodies (We get some vitamin B12 and K from
bacterial metabolism.)
C. Decomposition of dead organic material
1. Bioremediation (e.g., oil spills)
2. Sewage treatment
D. Production of antibiotics (e.g., streptomycin)
E. Production of cheese, yogurt, beer, wine, bread
IX. Harmful effects on other organisms
A. Some (relatively few) prokaryotes are pathogenic.
1. Pathogen - an organism that causes disease
2. In the late 19th century, physicians and biologists were starting to prove that some
diseases were caused by microbial agents.
B. German physician Robert Koch proposed a set of rules (Koch’s postulates) for testing
the relationship between a disease and a microorganism. (These rules were very
important in a time when it was not widely accepted that microorganisms cause
disease.)
1. The microorganism could always be found in the diseased individual
2. The microorganism taken from the host could be grown in pure culture
3. A sample of the culture produced the disease when injected into a new host
4. The newly infected host yielded a new pure culture
C. Invasiveness - ability of a pathogen to multiply within host
D. Toxigenicity - ability of a pathogen to produce toxic substances
1. Corynebacterium diptherinae, the agent that causes diphtheria, has low
invasiveness and multiples only in the throat, but its toxigenicity is so great that the
entire body is affected.
2. In contrast, Bacillus anthracis, which causes anthrax (a disease primarily of cattle
and sheep), has low toxigenicity but invasiveness that is so great that the entire
bloodstream ultimately teems with bacteria.
E. Some diseases caused by prokaryotes
1. Gram negative rods
a. Plague
b. Cholera (contaminated water)
c. Salmonellosis (Salmonella)
d. E. coli (some strains pathogenic)
e. Typhoid (water, food, etc.)
f. Crown gall (in plants)
2. Spirochetes
a. Syphilis
b. Lyme disease
3. Gram positive Bacteria
a. Botulism

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 b. Staph infections (Staphylococcus)
X. Domain Bacteria, Kingdom Eubacteria
A. Consists of 12 phyla (revision continues)
B. Selected groups (not necessarily monophyletic)
1. Cyanobacteria (blue-green "algae")
a. Photoautotrophs
b. Many fix nitrogen
c. Complex internal membranes
d. Very similar to the chloroplasts found in some Eukarya
2. Mycoplasmas
a. Smallest organisms
b. No cell wall
c. Can't be treated with penicillin and other antibiotics that attack cell wall
(1) Found in the bodies of animals, mycoplasmas live on the surfaces of the
mucous membranes
(2) Causes urinary tract infections and some forms of pneumonia.
XI. Domain Archaea, Kingdom Archaebacteria
A. Consists of 4 phyla (revision continues)
1. Hyperthermophiles (extreme thermophiles)
a. Live in very hot and sometimes very acidic environments
b. Example: sulfolobus and other members of this genus live in hot sulfur springs
at temperatures of 70-75C. They die of cold at 55C (131F). Hot springs are
also extremely acidic. This genus grows best at pH values between 2-3, but
tolerates levels to 0.9
2. Methanogens
a. Obligate anaerobes
b. Produce methane (ca. 2 billion tons/year)
c. Sole source of methane in our atmosphere
d. One species can live and grow at 110 C (current record)
(1) Produces methane by reducing carbon dioxide, which is a key step in their
energy metabolism
(2) Methanogens release about 2 billion tons of methane gas into the Earth's
atmosphere accounting for all the methane in our air.
(3) This includes the methane produced in mammalian flatulence. About 1/3
of methane production comes from methanogens in the guts of grazing
herbivores like cows.
3. Extreme halophiles
a. Live in very salty environments
b. Often appear pink due to carotenoids
c. Can live in very basic (pH 11) environments - if you view salt flats from
orbiting satellites, they often have a pink appearance due to these archaea.
4. Thermoplasma
a. No cell wall

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 b. Have heat-loving and acid-loving requirements; found in coal deposits.
c. It has the smallest genome among the Archaea and perhaps of all living
organisms.
XII. Eukaryotes
A. Domain Eukarya
B. Characteristics of eukaryotes
1. True nucleus (almost always)
2. Membrane-bound organelles
3. Usually much larger than prokaryotes
4. Some unicellular, some multicellular
5. Mitosis (and in many species meiosis) occurs
XIII. Protists
A. Domain Eukarya, Kingdom Protista
B. There have been several changes in how organisms in this group have been classified
C. Most protists were formerly classified into a diverse group called Protozoans that ingest
their food by endocytosis
D. Kingdom Protista is sort of a grab bag of organisms that don't fit well in the other
Kingdoms.
E. For the purposes of this class, Protists are all the eukaryotic organisms that are not
plants, fungi or animals.
F. Approximately 60,000 known species
G. General characteristics of protists
1. Most are unicellular, but some are multicellular
2. Most are aquatic, in fresh or salt water or in body fluids; some live in moist
terrestrial environments (e.g. slime molds inhabit damp soil and moist decaying
bark of rotting trees)
H. Metabolism
1. Protists are diverse in the structure, but not so diverse in their metabolism as
prokaryotes
2. They do have several different nutritional modes however
a. Some are photoautotrophs- there are several kinds of photosynthetic protists
that some biologists consider to be algae (alga singular)
b. Some are chemoheterotrophs
(1) Some absorb food (fungus-like; slime molds)
(2) Some ingest food (animal-like; amoeba)
c. Some are mixotrophic - can function as autotrophs or heterotrophs (euglena)
I. Locomotion
1. Few protist groups consist entirely of nonmotile forms. Most groups include
members that have the capability of locomotion
2. Flagella (flagellates) - moves like a whip
3. Pseudopodia (amoebas) - Amoebas are protists that form pseudopodia, extensions
of their constantly changes body (They are blob-like in nature.)
4. Cilia (ciliates) - tiny, hairlike organelles that beat in a coordinated fashion to move

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 forward or backward
J. Selected anatomical features
1. Contractile vacuoles- Protists that live in fresh water environments need to be able
to excrete excess water. Some have contractile vacuoles that remove water.
2. Food vacuoles - Protists such as Paramecium (a ciliate) engulf solid food by
endocytosis, forming a food vacuole in which food is digested.
3. Cell surfaces
a. Plasma membrane only (some amoebas) - a few protists, such as some
amoebas, are surrounded only by a plasma membrane, but most have stiffer
surfaces that maintain the structural integrity
b. Cell wall - present in many algae; diatoms have cell walls made with silicon
c. Pellicle (flexible protein covering) - formed by layers of membranes; varies in
structure and characteristics; present in Euglena
d. The protozoans, which lack a cell wall, have a variety of other ways of
strengthening their surface, including an exoskeleton (external shell):
(1) Exoskeleton produced by organism (foraminiferans -organic matrix
hardened with calcium carbonate)
(2) Exoskeleton constructed from materials in environment (some amoebas
use grains of sand)
K. Reproduction
1. Asexual (mitosis + cytokinesis) - includes binary fission (simple splitting of the
cells), multiple fission, budding (the outgrowth of a new cell from the surface of an
old one), and the formation of spores (resting structures that are very resistant to
harsh condition).
2. Sexual
a. In some protists, as in animals, the gametes are the only haploid cells
b. In other forms, such as algae, both diploid and haploid cells undergo mitosis
giving rise to alternation of generations (alternating generations of haploid
stages and diploid stages).
c. Some species exchange genetic material apart from reproductive process;
termed Protozoan Conjugation
L. Endosymbiosis - some protists live in other organisms, even in other protists
XIV. Classifications of protists
A. Diplomonadida and Parabasala
1. The most ancient groups of protists
2. Distinguished by their lack of mitochondria
3. All members of this group have flagella and 2 nuclei
4. Giardia lambilia (Diplomonadida)
a. A parasite that infects human intestines causing abdominal cramps and severe
diarrhea
b. Infects 2% of the human population worldwide
c. Usually associated with drinking water that is contaminated by human or other
vertebrate feces

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 5. Trichomonas vaginalis (Parabasala)
a. A parasite that inhabits the vagina in human females
b. If the normal acidity of the vagina is disturbed, it can proliferate and overcome
the beneficial microbial inhabitants.
B. Euglenozoa
1. Euglenoids
a. Euglena have an anterior pocket from which one or two flagella emerge and a
unique storage molecule (a glucose polymer) called paramylon.
b. Some Euglena have the unique ability of being mixotrophic, which describes
organisms that are capable of deriving metabolic energy both from
photosynthesis and from external energy sources.
2. Kinetoplastids
a. Distinguished by a single large mitochondrion associated with a kinetoplast,
which is a unique organelle that houses extranuclear DNA.
b. Trypanosoma is a parasite that causes sleeping sickness and is transmitted by
the tsetse fly. The parasite is successful because it constantly alters its
molecular structure and thus the host is not able to evolve defense against it.
C. Alveolata
1. Unicellular organisms with alveoli (subsurface cavities of which the function is
unknown).
2. Group contains three divisions (Dinoflagellates, Apicomplexa, and Ciliophora).
3. Dinoflagellates
a. Most are unicellular (few colonial)
b. Cell walls composed of cellulose which give them a rigid, armor-like
appearance.
c. Two flagella
d. “Red tide”, explosive blooms of dinoflagellates, color is produced from red
pigment in plastids; some produce toxins
e. Some bioluminescent forms which causes the florescent glow in the ocean at
night
4. Apicomplexa
a. Most are parasites on animals
b. Groups get its name from apex that contains a complex of specialized
organelles for penetrating its host
c. Plasmodium is a parasite that causes malaria; typically found in live and/or red
blood cells of its host
5. Ciliophora
a. The ciliated members of this group have a unique arrangement of DNA.
b. Large macronucleus has >50 copies of the genome and each micronuclei has
many copies of individual genes.
c. Reproduce through conjugation
D. Stramenophila
1. Hair-like projects on flagella distinguish this group from others.

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 2. Flagella is only present during motile reproductive stage
3. Most are unicellular, however some multicellular forms (algae)
4. Four divisions (Oomycota, Bacillariphyta, Chrysophyta, Phaeophyta)
5. Oomycota
a. Group originally classified in Kingdom Fungi because similar body form
(hyphae) and similar ecology.
b. Includes water molds, white rusts, and downy mildews
6. Bacillariphyta (Diatoms)
a. Unicellular with glass-like walls containing silica.
b. Photosynthetic
c. After death, diatoms leave a silica “skeleton”
7. Chrysophyta (Golden algae)
a. Appears golden in color as a result of carotene and xanthophyll pigments
b. Most are unicellular; few are colonial
c. Golden algae can be heterotrophic, photosynthetic, or mixotrophic
8. Phaeophyta (Brown algae)
a. Commonly known as kelp; includes the largest and most complex algae.
b. Multicellular and mostly marine
c. Characterized by the presence of a blade (leaf-like structure), stipe (stem-like
structure) and holdfast (root-like structure).
d. Alginic acid - used in ice cream, cosmetics - the cell walls of brown algae may
contain as much as 25 percent alginic acid, which is a gummy polymer of sugar
acids that acts as a glue.
E. Rhodophyta (Red algae)
1. Name is for clade and for division
2. Multicellular are usually reddish in color which is the result of a pigment
phycoerythrin (only in red algae)
3. Often found in deeper depths of the ocean compared to other algae.
4. Identified by branched body form and lack of structures found in Brown algae.
F. Chlorophyta (Green algae)
1. Name is clade and division
2. Distinguished by their “plant green” color
3. Share same pigments (chlorophyll a and b) as plants
4. 7000 species have been described; very diverse
5. Unicellular, colonial and multicellular forms
G. Mycetozoa (Slime molds)
1. Similar to fungus
2. Use pseudopodia (like amoebas) for movements
3. Decomposers
H. Protists of unknown ancestry
1. Rhizopoda (Amoebas)
a. Lobe-like pseudopodia
2. Actinopoda (Radiolarians)

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 a. Ray-like pseudopodia
b. Have exoskeletons of silicon
3. Foraminfera
a. Porous shells with pseudopodia extending through shells
b. Exoskeleton hardened by CaCO3 which is shed during reproduction
c. The discarded skeletons of ancient foraminiferans make up extensive limestone
deposits in various parts of the world, forming a covering hundreds to
thousands of meters deep on the ocean bottom (White Cliffs of Dover)

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