NPTEL – Biotechnology – Microbiology

Module 8 – Diversity of Microbial World

Lecture 1- Microbial Taxonomy and Phylogeny
Introduction
Living organisms are fascinating by its diversity whether it is plants, animals or
microbes. A handful of soil is populated with more than the human population on earth.
They play important essential roles in nature. So if we arrange these microbes in order or
hierarchy by based on its similarity or differences in any characteristics, we can easily get
to know and get easy access to all the microbes. So it is desirable to determine the
classification. Greek Philosopher Aristotle who is the one classified the living things as
plants and animals around 2000 years ago. So in this lecture, we will learn about
taxonomy, how is it classifified? What methods are available to classify them? And then
brief description about microbial evolution and diversity and its phylogeny.
Taxonomy
Taxonomy [Greek taxis, arrangement, and nomos, law, or nemein, to distribute] is
defined as the science of biological classification. In simple term, taxonomy is orderly
arranging organisms under study into groups of larger units. It consists of three
interrelated parts namely
1. Classification is the arrangement of organisms into groups or taxa (s., taxon) based
on mutual similarity or evolutionary relatedness.
2. Nomenclature is concerned with the assignment of names to taxonomic groups in
agreement with published rules.
3. Identification is the practical side of taxonomy, the process of determining that a
particular isolate belongs to a recognized taxon. (So in short Identify-Naming them
and classify them)
Classification
It is bringing order to the diverse variety of organisms present in nature. So
there are two general ways the classification can be constructed. First one is based on the
morphological characters (phenetic classification) and second is based on evolutionary
relationship (phylogenetic classification)
Phenetic classification- Grouping organisms together based on the mutual
similarity of their phenotypic characteristics. It does not provide information
about phylogenetic relations.
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Phylogenetic classification- These are systems based on evolutionary
relationships rather than external appearance (the term phylogeny [Greek phylon,
tribe or race, and genesis, generation or origin] refers to the evolutionary
development of a species). It is based on the direct comparison of genetic
materials and/or gene product.

Nomenclature (Binomial system)

Biologists in the middle ages used to follow polynomial system, i.e naming
organisms with many names (poly-many, nomo- name). For example name for the
European honeybee, was Apis pubescens, thorace subgriseo, abdomine fusco, pedibus
posticis glabris utrinque margine ciliatis (just for example no need to be memorized).
Later Binomial systems were developed by Swedish biologist Carolus Linnaeus (1707–
1778) based on the anatomical characteristics of plants and animals. Nomenclature in
microbiology is developed based on the principals established for the plant and Animal
kingdom by Linnaeus. The first word in the binomial is the genus name and is always
capitalized. The second word is species name and never capitalized. For example
honeybee, Apis mellifera

Taxonomic ranks:
In prokaryotic taxonomy the most commonly used levels or ranks (in ascending order)
are species, genera, families, orders, classes, phyla, kingdom or domain. In order to
remember the seven categories of the taxonomic hierarchy in their proper order, it may be
useful to memorize a phrase such as “kindly pay cash or furnish good security”
(kingdom–phylum– class–order–family–genus–species). The basic taxonomic group in
microbial taxonomy is the species.
A species is a collection of strains that have a similar G+C composition and 70% or
greater similarity as judged by DNA hybridization. Ideally a species also should be
phenotypically distinguishable from other similar species. An example of hierarchy in
taxonomy is given below.

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Rank Example
Domain Bacteria
Phylum Proteobacteria
Class γ- Proteobacteria
Order Enterobacteriales
Family Enterobactericeae
Genus Shigella
Species S.dysentriae

A strain is a population of organisms that is distinguishable from at least some

other populations within a particular taxonomic category. It is considered to have
descended from a single organism or pure culture isolate. Strains within a species may
differ slightly from one another in many ways. Biovars are variant prokaryotic strains
characterized by biochemical or physiological differences, morphovars differ
morphologically, and serovars have distinctive antigenic properties. One strain of a
species is designated as the type strain. It is usually one of the first strains studied and
often is more fully characterized than other strains; however, it does not have to be the
most representative member but this strain can be considered as reference strain and can
be compared with other strains. Each species is assigned to a genus, the next rank in the
taxonomic hierarchy. A genus is a well-defined group of one or more species that is
clearly separate from other genera.

Techniques for identifying or determining taxonomical characters

In order to identify and classify microorganisms, we need to know about their

characteristics. There are two ways to determine the taxonomical characters; classical
and molecular characters

Classical characteristics:- This approach uses morphological, biochemical,

physiological, ecological and genetic characteristics. It is mainly used in microbial
taxonomy.

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1. Morphology:- Morphology is the one which can be easily studied and analyzed.
Structural features (cell shape, size, colony morphology, appendages, and etc.)
depend on the expression of many genes, are usually genetically stable.
2. Physiology and metabolism:- Organisms are classified based on the
requirements for growth characters like carbon and nitrogen sources, cell wall
constituents, general nutritional type, energy sources, optimum growth
temperature, Motility.
3. Ecology:- These are taxonomically valuable because even very closely related
microorganisms can differ considerably with respect to ecological characteristics.
The ability to cause disease in a particular host; and habitat preferences such as
requirements for temperature, pH, oxygen, and osmotic concentration are
examples of ecological characteristics.
4. Genetic analysis:- The study of chromosomal gene exchange between species
through transformation and conjugation (in Enteric bacteria) is sometimes useful
in their classification. Most bacteria are harboring plasmids, classification based
on plasmid is also an important part of classification.
Molecular characteristics:-This is the most powerful approaches to study taxonomy by
analyzing proteins and nucleic acids. Because these are either direct gene products or the
genes themselves, comparisons of proteins and nucleic acids yield considerable
information about true relatedness.
1. Comparing amino acid sequences:- Comparison of amino acid sequences of proteins
from different organisms reveals its taxonomic relations. The most direct approach is to
determine the amino acid sequence of proteins with the same function. If the sequences
of proteins with the same function are similar, the organisms possessing them are
probably closely related. The electrophoretic mobility of proteins is useful in studying
relationships at the species and subspecies levels. Antibodies can discriminate between
very similar proteins, and immunologic techniques are used to compare proteins from
different microorganisms.
2. Nucleic acid composition: By direct comparison of microbial genomes and based on
the G+C content of different organisms (Escherichia coli 48-52 %). And genomic
fingerprinting (RFLP, AFLP) reveals its relatedness with others.

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3. Nucleic acid hybridization: It uses the property of complementarities in double
stranded DNA. More distantly related organism can be identified based on DNA-RNA
hybridization
4. Nucleic acid sequencing: Techniques are now available to sequence both DNA and
RNA. 5S and 16S RNA (prokaryotes), 18S (fungi) analysis of microorganisms can reveal
their relatedness because of its functional role is same in all ribosomes and slow
structural changes with time.

Microbial evolution and Diversity

It has been estimated that our planet is about 4.6 billion years old. Around 3.5 to 3.8
billion years old fossilized remains of prokaryotic cells have been discovered in
sedimentary rocks. Thus earlier prokaryotes were anaerobic and arose shortly after the
earth cooled. Cyanobacteria and oxygen-producing photosynthesis probably developed
2.5 to 3.0 billion or more years ago.
It appears likely that modern eukaryotic cells arose from prokaryotes about 1.4 billion
years ago.
Two hypotheses for the evolution of eukaryotic cells
1. Organelles arose within prokaryotes from the invagination of the plasma membrane
2. Endosymbiotic hypothesis
Fusion of ancient true bacteria and archaea to form a nucleus. They proposed that the
eukaryotic line diverged from the Archaea and then the nucleus formed, possibly from
the Golgi apparatus
Mitochondria and chloroplasts develop later from a permanent symbiotic relationship
with other bacteria, e.g., cyanelle (cyanobacterium) living inside the protist Cyanophora
paradoxa
Cyanobacteria have been considered the most likely ancestors of chloroplasts. More
recently Prochloron has become the favorite candidate. The existence of this bacterium
suggests that chloroplasts arose from a common ancestor of prochlorophytes and
cyanobacteria. Mitochondria arose from an endosymbiotic relationship between the free-
living primitive eukaryotic and bacteria with aerobic respiration (possibly an ancestor of
three modern groups: Agrobacterium, Rhizobium, and Rickettsia).

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Divisions of Life

Kingdom systems of classification

-Five-kingdom system (Whittaker, 1960s) - based upon cell type, organization, and the
means of nutrient acquisition (Monera, Protista, Fungi, Plantae, Animalia)
-Six-kingdom system - differs from five-kingdom system by dividing prokaryotes into
bacteria and archaea (Bacteria, Archaea, Protista, Fungi, Plantae, Animalia)
-Eight-kingdom system (Cavalier-Smith) - further division of the protists using rRNA
data and grouping organisms into two empires (Eucaryota and Bacteria) containing a total
of eight kingdoms [(Bacteria, Archaea), (Archezoa, Protista, Plantae, Chromista, Fungi,
Animalia)]

Domains

Advances in genomic DNA sequencing of the microorganisms, biologists are

increasingly adapting the classification of living organisms that recognizes three
domains, a taxonomic level higher than kingdom. Archaebacteria are in one domain,
eubacteria in a second, and eukaryotes in the third. Domain Eukarya is subdivided into
four kingdoms plants, animals, fungi, protists.

Fig. 1 Three domains based on Woese rRNA sequence analysis

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Domain- Archaebacteria
The term archaebacteria (Greek, archaio, ancient) refers to the ancient origin of
this group of bacteria, which seem to have diverged very early from the eubacteria. They
are inhabited mostly in extreme environments. The archaebacteria are grouped (based
primarily on the environments in which they live) into three general categories
methanogens, extremophiles and non extreme Archaebacteria.

Fig. 2 - Universal Phylogenetic Tree

Domain- Bacteria

The Eubacteria are the most abundant organisms on earth. It plays critical roles
like cycling carbon and sulfur. Much of the world’s photosynthesis is carried out by
eubacteria. However, certain groups of eubacteria are also responsible for many forms of
disease.

Domain- Eukarya

It consists of four kingdoms. The first of which is protista, mostly unicellular organism
like amoeba. The other three kingdoms are plants, fungi, animals. Multicellularity and
sexuality are the two unique characters that differentiate from prokaryote and eukaryotes.

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Fig. 3. Phylogenetic tree. a) unrooted tree, b) rooted tree.

Ways to determine the phylogenetic relationships

Molecular chronometers

This concept, first suggested by Zuckerkandl and Pauling (1965), which is based on
thought that the sequences of many rRNAs and proteins gradually change over time
without destroying or severely altering their functions. Changes increases with time
linearly. If sequences of similar molecules from two organisms differs, it means that they
diverged very long time ago.

Phylogenetic tree
Phylogenetic relationships are illustrated in the form of branched diagrams or
trees (denrograms). A phylogenetic tree is a graph made of branches that connect nodes.
The nodes represent taxonomic units such as species or genes; the external nodes, those
at the end of the branches, represent living organisms. The tree may have a time scale, or
the length of the branches may represent the number of molecular changes that have
taken place between the two nodes. Finally, a tree may be unrooted or rooted.
An unrooted tree simply represents phylogenetic relationships but does not
provide an evolutionary path. Figure 3. a. shows that A is more closely related to C than
it is to either B or D, but does not specify the common ancestor for the four species or the
direction of change. In contrast, the rooted tree Figure 3. b. does give a node that serves
as the common ancestor and shows the development of the four species from this root.

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Parsimony analysis

Phylogenetic relationships also can be estimated by techniques such as parsimony

analysis. In this approach, relationships are determined by estimating the minimum
number of sequence changes required to give the final sequences being compared. It is
presumed that evolutionary change occurs along the shortest pathway with the fewest
changes or steps from an ancestor to the organism in question.
Oligonucleotide signature sequences
The 16S rRNA of most major phylogenetic groups has one or more characteristic
nucleotide sequences called oligonucleotide signatures. Oligonucleotide signature
sequences are specific oligonucleotide sequences that occur in most or all members of a
particular phylogenetic group. They are rarely or never present in other groups, even
closely related ones. Thus signature sequences can be used to place microorganisms in
the proper group.

Polyphasic taxonomy

Studying phylogeny based on both genotypic and phenotypic information ranging

from molecular characteristics to ecological characters.

Numerical Taxonomy

Computer based approaches of grouping organisms is called Numerical taxonomy

which is based on presence or absence of selected characters in the group of organisms. It
is method of estimating percent similarity (ratio between the number of characters same
and total number of characters organisms having). This method has great practical
usefulness as well as being relatively unbiased in its approach. It has high degree of
stability and predictability.

REFERENCES:

Text Books:

1. Jeffery C. Pommerville. Alcamo’s Fundamentals of Microbiology (Tenth Edition).

Jones and Bartlett Student edition.

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2. Gerard J. Tortora, Berdell R. Funke, Christine L. Case. Pearson - Microbiology: An
Introduction. Benjamin Cummings.

Reference Books:

1. Lansing M. Prescott, John P. Harley and Donald A. Klein. Microbiology. Mc Graw

Hill companies.

2. Biology, Raven and Jhonson, 6th edition (2001)

3. Microbiology, Pelczar. M.J , Chan E.C.S, Kreig N.R, 5th edition (2007)

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Module 8 – Diversity of Microbial World

Lecture 2 – Archaea and Bacteria
Archaea

It can be described as, group of (primitive) bacteria living in extreme aquatic and
terrestrial environments. They can stain either gram positive or gram negative and may be
spherical, rod-shaped, spiral, plate-shaped, or pleomorphic. Some are single cells,
whereas others form filaments or aggregates.

They range in diameter from 0.1 to over 15 μm, and some filaments can grow up to 200
μm in length. Multiplication may be by binary fission, budding, fragmentation, or other
mechanisms.

Cell walls

Archaeal cell wall is made of glycoprotein or protein instead of peptidoglycan.

Methanobacterium and some other methanogens have walls containing pseudomurein, a
peptidoglycanlike polymer that has L-amino acids in its cross-links, N-
acetyltalosaminuronic acid, and β(1→3) glycosidic bonds Methanosarcina and
Halococcus lack pseudomurein and contain complex polysaccharides similar to the
chondroitin sulfate of animal connective tissue. Other heteropolysaccharides are also
found in gram- positive walls.

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Lipids and membranes

This is the most distinctive nature of archaea is membrane lipids. They are different from
bacteria and eukaryotes. Archaeal lipids are derivatives of isopranyl glycerol ethers rather
than the usual glycerol fatty acid esters. They also contain phospholipids, sulfolipids and
glycolipids. Archaeal nonpolar membranes are the derivative of squalene, 30 carbon
compound, presence of diethers, tetraethers are needed for their stability to thrive in
extreme environments.

Fig. 4. Archael glycerolipids

Metabolism

Archaeal metabolism varies greatly between the members of different groups. They are
just as diverse physiologically. They can be aerobic, facultatively anaerobic, or strictly
anaerobic. Nutritionally they range from chemolithoautotrophs to organotrophs.
Archaeal carbohydrate metabolism is best understood. The enzyme 6-
phosphofructokinase has not been found in archaea, and they do not appear to degrade
glucose by way of the Embden-Meyerhof pathway. Extreme halophiles and thermophiles
catabolize glucose using a modified form of the Entner-Doudoroff pathway in which the
initial intermediates are not phosphorylated. In contrast with glucose degradation,
gluconeogenesis proceeds by a reversal of the Embden-Meyerhof pathway in halophiles
and methanogens. All archaea that have been studied can oxidize pyruvate to acetyl-CoA.
They lack the pyruvate dehydrogenase complex present in eucaryotes and respiratory
bacteria and use the enzyme pyruvate oxidoreductase for this purpose. Halophiles and the
extreme thermophile Thermoplasma do seem to have a functional tricarboxylic acid
cycle.

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Archaeal Taxonomy

Based on the rRNA data, it has been divided into two major phylum namely
Euryarchaeota, Crenarchaeota. On the basis of morphology and physiology, it has been
divided into five different major groups. (1. Methanogenic archaea, 2.Archaea sulfate
reducers, 3.Extremely halophilic archaea, 4. Cell wall–less archaea, 5.Extremely
thermophilic S0-metabolizers).

1. Phylum-Euryarchaeota (Eures: wide)

The euryarcaeota are given this name because they occupy many diverse ecological
niches and have a variety of metabolic patterns. It consists of seven classes
(Methanobacteria, Methanococci, Halobacteria, Thermoplasmata, Thermococci,
Archaeglobi, and Methanopyri), nine orders and 15 families.

Methanogens

They are strict anaerobes that obtain energy through the conversion of CO2 and H2 into
methane. This is the largest group of archaea. There are five orders (Methanobacteriales,
Methanococcales, Methanomicrobiales, Methanosarcinales, and Methanopyrales) and 26
generas. Methanogens thrive in anaerobic environments rich in organic matter: the
rumen and intestinal system of animals, freshwater and marine sediments, swamps and
marshes, hot springs, anaerobic sludge digesters, and even within anaerobic protozoa.
Rumen methanogens are so active that a cow can belch 200 to 400 liters of methane a
day.

Halobacteria

They are aerobic heterotrophs with respiratory metabolism and require complex nutrients,
usually proteins and amino acids, for growth. Species are either nonmotile or motile by
lophotrichous flagella. The extreme halophiles, class Halobacteria, are another major
group of archaea, currently with 15 genera in one family, the Halobacteriaceae. The most
obvious distinguishing trait of this family is its absolute dependence on a high
concentration of NaCl. These procaryotes require at least 3 to 4 M NaCl (17 to 23%) for
their growth optimum. Halobacterium produces energy by trapping light and synthesize
ATP with the help of rhodopsin.

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Thermoplasms

Thermoacidophiles are the characteristics of lacking cell walls. At present, only two
genera, Thermoplasma and Picrophilus, are known. Thermoplasma grows in refuse piles
of coal mines. These piles contain large amounts of iron pyrite (FeS), which is oxidized
to sulfuric acid by chemolithotrophic bacteria. As a result the piles become very hot and
acidic. This is an ideal habitat for Thermoplasma since it grows best at 55 to 59°C and
pH 1 to 2.

Picrophilus is grows at sulfur fields with optimum pH requirement of less than 1. They
are aerobic, irregular shaped cocci, having sulfur layer outside its plasma membrane.

Extremely Thermophilic S-Metabolizers

This physiological group contains the class Thermococci, with one order,
Thermococcales. The Thermococcales are strictly anaerobic and can reduce sulfur to
sulfide. They are motile by flagella and have optimum growth temperatures around 88 to
100°C. The order contains one family and two genera, Thermococcus and Pyrococcus.

Sulfate-Reducing Archaea

Archaeal sulfate reducers are found in the class Archaeoglobi and the order
Archaeoglobales. This order has only one family and one genus. Archaeoglobus contains
gram-negative, irregular coccoid cells with walls consisting of glycoprotein subunits
Archaeoglobus is extremely thermophilic.

2. Phylum- Crenarchaeota

They are the extreme hyperthermophiles which needs optimum temperature of 105 ⁰C for
their growth.At present, the phylum contains 69 genera; two of the better-studied genera
are Thermoproteus and Sulfolobus. Thermoproteus is a strict anaerobe and grows at
temperatures from 70 to 97°C and pH values between 2.5 and 6.5. It is found in hot
springs and other hot aquatic habitats rich in sulfur. Sulfolobus are aerobic, with a
temperature optimum around 70 to 80°C and a pH optimum of 2 to 3.

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Bacteria

Bacteria are prokaryotes, evolved first on living earth. They does not contain membrane
bound nucleus and organelles. Almost all the bacteria are having circular genome and
extrachromasomal DNA which helps in survive different environments. Bacteria
reproduce by prokaryotic fission, resulting in two genetically identical daughter cells.
Most of the bacteria are unicellular in nature but sometimes they form aggregates. The
most common shapes of bacteria are spheres (cocci), rods (bacilli), spirals.

Bacterial cell walls are made of peptidoglycan contains sugar moieties (N-
acetylgluscosamine, and N-acetylmuramic acid cross linked with pentapeptide (D-
aminoacids). Gram stain is a valuable tool to identify the bacteria based on the cell wall
constituents. Gram-positive bacteria have simple cell walls with large amounts of
peptidoglycan and which retains crystal violet. Gram-negative bacteria have more
complex cell walls with less peptidoglycan and which retains saffron, the counter strain.
Presence of lipid layer is a unique characteristic of Gram negative bacteria. Most of the
gram negative bacteria are causative agent of many human diseases than Gram positive
bacteria. The lipopolysaccharides on the walls of gram-negative bacteria are often toxic,
and the outermembrane protects the pathogens from the defenses of their hosts.

Capsules are the slimy layer produced by most of the bacteria which helps them to adhere
together and form colonies. Two kinds of filamentous structures may be attached to the
cell wall: The bacterial flagellum rotates like a propeller to pull the cell along while
movement. Pili help bacteria attach to one another in conjugation, and fimbriae help
them attach to surfaces. Many prokaryotes are capable of taxis, movement toward
nutrients or oxygen (positive chemotaxis) away from a toxic substance (negative
chemotaxis). Some bacteria form resistant cells called endospores when an essential
nutrient is lacking in the environment and it may remain dormant but viable for centuries
or longer.

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Classification based on nutrition and metabolism

Organisms can be categorized by their nutrition, based on how they obtain energy and
carbon to build the organic molecules that make up their cells. Organisms that obtain
energy from light are phototrophs. Organisms that obtain energy from chemicals in their
environment are chemotrophs. Organisms that need only an inorganic compound such as
CO2 as a carbon source are autotrophs. Organisms that require at least one organic
nutrient—such as glucose—as a carbon source are heterotrophs. Based on requirement of
oxygen they are classified as obligate aerobes (requires O2 for respiration), facultative
anaerobes (can grow both aerobically and anerobically), and obligate anaerobes (does not
require O2).

Bacterial taxonomy

Until the late 20th century, biologists based prokaryotic taxonomy on criteria such as
shape, motility, nutritional mode, and Gram staining. Although these criteria may be
valuable in culturing and identifying pathogenic bacteria, they may not reflect
evolutionary relationships. Applying molecular data to the investigation of prokaryotic
phylogeny has been very fruitful. Microbiologists began comparing sequences of
prokaryotic genes in the 1970s. Carl Woese and his colleagues used ribosomal RNA
(rRNA) as a marker for evolutionary relationships.
In 1923, David Bergey and colleagues set out to publish a definitive book on the
identification and classification of bacteria. A Survey of Bacterial Phylogeny and
Diversity - based on the 2nd edition of Bergey's
Volume 1: The Archaea, Cyanobacteria, Phototrophs and Deeply Branching Genera
Archaea - divided into two kingdoms
a. Crenarchaeota - diverse kingdom that contains thermophilic and hyperthermophilic
b. Euryarchaeota - contains primarily mathanogenic and halophilic bacteria and also
Eubacteria - complex with several small groups of phototrophs, cyanobacteria, and
deeply branching eubacteria

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Based on Bergeys’ manual Domain Bacteria contains six phyla in volume 1
• Phylum Aquificiae- earliest branch of bacteria that contain autotrophs which
utilize hydrogen for energy production
• Phylum Thermotogae - anaerobic, thermophilic, and fermentative Gram negative
bacteria
• Phylum “Deinococcus Thermus” - radiation resistant bacteria
• Phylum Chloroflexi- green non-sulfur bacteria that carries out anoxygenic
photosynthesis
• Phylum Cyanobacteria - oxygenic photosynthetic bacteria
• Phylum Chlorobi - green sulfur bacteria that carry out anoxygenic photosynthesis
Volume 2 - Gram negative proteobacteria (purple bacteria)
Based on the nutritional type and rRNA data Gram negative- Proteobacteria have been
classified into five classes.
 Alphaproteobacteria-oligotrophic forms including the purple nonsulfur
photosynthesizers
 Betaproteobacteria - metabolically similar to alphaproteobacteria
 Gammaproteobacteria - diverse methods of energy metabolism
 Deltaproteobacteria - includes predators and the fruiting myxobacteria
 Epsilonproteobacteria - contains pathogens

Volume 3 - Gram positive bacteria with low G + C content (< 50%)

Three classes of the phylum Firmicutes
 Clostridia - tend to be anaerobic and endospore formers
 Mollicutes - mycoplasmas (no cell walls)
 Bacilli - Gram-positive aerobes or facultative anaerobes, rods or cocci, some
endospore formers
Volume 4 - Gram positive bacteria with high G + C content (> 50-55%)
 All belong to the phylum Actinobacteria
 Some are filamentous

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Volume 5 - Gram negative with various morphologies
Nine phyla of which four are presented below
• Phylum Planctomycetes - some have a membrane-bound nucleus
• Phylum Chlamydiae - obligate intracellular parasites important in disease
• Phylum Spirochaetes - helical-shaped, Gram-negative motile bacteria (axial
filaments)
• Phylum Bacteroidetes - ecologically significant species are found in this phylum
REFERENCES:

Text Books:

1. Jeffery C. Pommerville. Alcamo’s Fundamentals of Microbiology (Tenth Edition).

Jones and Bartlett Student edition.

2. Gerard J. Tortora, Berdell R. Funke, Christine L. Case. Pearson - Microbiology: An

Introduction. Benjamin Cummings.

Reference Books:

1. Lansing M. Prescott, John P. Harley and Donald A. Klein. Microbiology. Mc Graw

Hill companies.

2. Biology, Raven and Jhonson, 6th edition (2001)

3. Microbiology, Pelczar. M.J , Chan E.C.S, Kreig N.R, 5th edition (2007)

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Module 8 – Diversity of Microbial World

Lecture 3 – Fungi, algae, protozoa and slime molds

Fungi

Basic features:

It belongs to the domain Eukarya. They are unicellular (yeast) as well as multicellular
organism (filamentous fungi). Multicellular fungi are composed of filaments called
hyphae (singular: hypha). Hyphae may contain internal cross walls, called septa, which
divide the hyphae into separate cells. The hyphae may be branched. A mass of hyphae
that is not a reproductive structure is called a mycelium. Fungi are saprophytic; absorb
nutrients after degrading the organic matter and heterotrophs; require organic
compounds. They have cell walls composed of chitin. The hyphae of some symbiotic
fungi become specialized for penetrating the cells of the host. These hyphae are called
haustoria. Reproduce both sexually and asexually, typically through the production of
spores. Sexually produced spores are resting spores. In general, the life cycle involves the
fusion of hyphae from two individuals, forming a mycelium that contains haploid nuclei
of both individuals. The fusion of hyphae is called plasmogamy. The fused hyphae
containing haploid nuclei from two individuals are heterokaryotic. In some cases,
plasmogamy results in cells with one nucleus from each individual. This condition is
called dikaryotic. It has been classified based on the mode of reprodouction

 Lower fungi-

Fungi belongs to this family are having nonseptate walls and spores contained in
small sporangia. These classes of fungi include three groups: Chytridiomycota,
Oomycota, Myxomycota

 Higher fungi - true fungi (Kingdom Fungi)

Fungi belongs to this family are having septate cross walls
spores contained in complex structures
Include three groups: Zygomycota, Ascomycota, Basidiomycota

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Asexual Spores
Produced by mitosis and cell division
1. Sporangiospore
 Spores form sac called sporangium
 Sporangium forms at end of aerial hyphae called a sporangiophore. Hundreds of
sporangiospores in a single sporangium
2. Conidiospore
 Spores produced at the end of an aerial hyphae is called as conidiophore
 Conidia: chains of conidiospores on conidiophores
Sexual Spores
Sexual spores formed by fusion of two haploid nuclei into single diploid zygote. zygote
then undergoes meiosis to generate haploid spores (usually multiples of four)
1. Zygospores
One thick spore between two parent hyphae
2. Ascospores
Four spores in a sac called an ascus, at the end of one hyphae
3. Basidiospores
Four spores on the end of a basidium
Fungi organized into three Phyla based on the type of sexual spore:
Phylum Chytridiomycota (chytrids or water molds)
Basic features:
Aquatic, unicellular and coenocytic (multinucleate), hyphae are typically haploid (but
some diploid) - typically composed of a microscopic sphere, cell walls are made of chitin.
They have Rhizoids to penetrate food source. Many are parasitic on plants and other
fungi. Some are saprophytics.

Reproduction:
Asexual reproduction -Sporangium with single nucleus that splits off to produce a
flagellate zoospore with one flagellum. Sexual production through the formation of
sporophyte.

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Phylum Oomycota (water molds & mildews)

Basic features:
Aquatic, extensive nonseptate mycelium (unicellular, coenocytic), hyphae are diploid,
cell walls made of cellulose, they are heterotrophic: parasites on fish, plant pathogen, also
initial decomposers of dead insects.

Asexual Reproduction:
Hyphae grow, terminal portions of a mycelium pinch off to produce Zoospores, each
with two flagella.

Sexual Reproduction

Reproductive cells produced by meiosis several eggs per Oogonium, Antheridium

long and skinny (clavate).

Phylum Myxomycota (plasmodial slime molds)

Basic features:
They are terrestrial; contain no cell walls, vegetative structure are called as
plasmodium which is diploid (coenocytic), Amoeboid-feed by phagocytosis.
With adverse environmental conditions (ex. drought)-formation of a hardened
Sclerotium (multicellular resting structure)

Sexual Reproduction

Sporangium development -- meiosis to form 4 haploid spores, 3 of the 4 spores

disintegrate before release. Germination into amoeboid cells, some cells become
flagellate, fusion to form a zygote which later develops into a new plasmodium

Phylum Zygomycota (pin and bread molds)

Basic features:
Terrestrial, hyphae haploid, septate, cell walls made of chitin
Saprophytic
Produced complex reproductive structure; zygosporangium with one zygospore

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Asexual Reproduction:
Nonmotile spores on aerial sporangia
Spores are air dispersed
Sexual Reproduction
When two hyphae come in contact, they produce Gametangia by the initiation of
process of conjugation (fusion), and two haploid nuclei into a common cell and fertilize
with a thickened wall; Zygosporangium

Phylum Ascomycota (cup or sac fungi)

Include: morels, truffles, yeasts, dutch elm disease, corn blight

Basic features:
Hyphae are septate and monokaryotic (having one nucleus per compartment)
Produce complete reproductive structure called ascocarp with 8 ascospores

Asexual Reproduction:
Produce a sporangium-like conidium, within it called as conidiospores
Some species - no sexual reproduction seen, produce only conidia:
a) Penicillium - flavoring in cheese (blue, Roquefort, Camembert)
b) Aspergillus - aid in fermentation of soybean to produce Tofu

Sexual Reproduction

Hypal fusion leads to production of dikaryotic cells which in turn form into ascocarp. and
through meiosis it produces 8 ascospores.

Three types of sporocarps:

1) Apothecium - most common, cup fungi, morels
2) Perithecium - small flask-shape with small opening
3) Cleistothecium - no opening, release by decomposition

Yeasts: most common Saccharomyces

Yeast are single celled having diploid nucleus, mostly reproduce asexually by budding
Sexual reproduction through meiosis to from 4 ascospores.

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Phylum Basidiomycota (club fungi)
Include: mushrooms, coral fungi, rusts, smuts

Basic features:
Mainly terrestrial
Hyphae are septate and monokaryotic (having one nucleus per compartment)
Produce complete reproductive structure called basidiocarp with 4 basidiospores

Sexual Reproduction
Sexual process similar to that of ascomycetes
Fusion to get dikaryotic mycelia which develop into sporocarp-Basidiocarp
Immediate meiosis to form 4 Basidiospores on a Basidium (club-like structure)

Symbiosis Involving Fungi

Two types involving fungi:

1) mutualism - both species benefit
2) parasitism - one benefits, one harmed

Mutualism
Two examples: mycorrhizae & lichens

Mycorrhizae are the result of fungi in the roots of vascular plants

Fungus benefits: obtains photosynthates (esp.: sugar)
Plant benefits: obtains minerals (esp.: N, P)

Two types of mycorrhizae based on type of infection

1) Ectomycorhhizae (sheathing)
Grow between root cells of epidermis & cortex, not into cells, not beyond
endodermis
results in short, stubby roots
most common in: conifers, oaks, willows which are infected with basidiomycetes.

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2) Endomycorhhizae (internal)

Fungal hyphae grow into root cells

Within cell walls, NOT cell membrane
Mainly in epidermal & cortex cells
Most common in angiosperms (ex.: tulip tree) which are infected by zygomycetes

Lichens are the result of a fungus and an algae living together

a) The mycobiont - a fungus

Mostly ascomycetes, but some basidiomycetes
Provides a suitable environment & minerals to algae
b) The photobiont - an algae
An algae (green) or cyanobacterium
Provide carbohydrate & nitrogen compounds to fungus
symbiosis allows for them to live in very harsh environments: rock surfaces, tree
trunks
Ability to survive related to ability to dehydrate quickly
Fungal surface blocks UV light

Algae

Algae are photoautotrophic, unicellular (colony) as well as multicellular (filamentous).

Cell walls are made of cellulose or pectin, and require high moisture for their growth.
Reproduction is through sexual as well as asexual.

Five phyla

1. Brown algae
They are dark pigment producing, non motile multicellular organism, contains
chlorophyll a and b. Example. Sea weed.
2. Red alage
They are red pigment producing, non motile multicellular organism, contains chlorophyll
a and d. Example- Sea weed.

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3. Green Algae
They are filamentous unicellular as well as multicellular organisms, contains chlorophyll
a and b.
Example- Pond scum
4. Diatoms
They are light brown pigment producing unicellular organisms. Cell wall is made of
pectin and silicon oxide.
5.Dinoflagellates
They are unicellular, flagellated organisms. Their cellulose walls are interlocked.
Example- Plankton, red tide

Protozoa

Most of the protozoa are unicellular, aerobic, and chemoheterotrophic in nature.

Reproduction is through sexual as well as asexually. They also require high moisture for
their growth as algae. They have specialized structures to take food. Protozoa usually
covered with pellicle. Digestion occurs in vacuoles. Their life cycle switch between two
forms; one is trophozoite (vegetative and growing stage), and another one is cyst stage.
This is the survival stage for protozoa where they will move from one host to another,
resisting to the different environmental conditions. Cyst will turn back to their vegetative
stage when it finds favorable conditions.

Based on gene sequencing and motility it has been grouped into five major phyla
1. Archaeoa
Archaeoa are spindle shaped, lack mitochondria. They are having flagella at the front
end and common symbionts in animal. Example- Giardia
2. Apicomplexa
Organisms present in this groups are obligate intracellular parasites, and non motile in
mature form. Usually transmitted by insects, and having complex life cycle with different
stages in different hosts. Example- Plasmodioum (malaria)
3. Amoebozoa
This group contains causative agent of dysentery, and they move with the help of
pseudopods. Example- Entamoeba

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4. Ciliophora
Only one pathogen in group called Balantium coli, which is also a causative agent of
dysentery. They move with the help of cilia present on the surface of cell. Example-
Paramacium.
5. Euglenozoa
Asexual mode of reproduction and movement with the help of flagella called
zooflagellates. It contains two groups; a) Euglenoids: they are photoautotroph as well as
chemoautotrophs, has chlorophyll a, movement via flagella, b) Hemoflagellates: they are
long slender cells with undulating membrane and flagellum. Transmitted through insects
and live in host blood as the name implies. Example- Trypansoma

Slime Molds

They are having both the properties of fungi and amoeba, and mostly related to
amoebazoa. They are the parasites of bacteria and fungi and produce spores in
unfavorable conditions. It has been divided into two phyla; cellular slime molds and
plasmodial slime molds.
1. Cellular slime molds
In favorable conditions, they exist as unicellular amoeba and in unfavorable conditions;
they form as aggregate of multicellular mushroom like structure to generate spores. When
return to favorable conditions, spores germinate into unicellular amoeba.

2. Plasmodial slime molds

In favorable conditions, they exist as plasmodium containing multinucleated mass of
protoplasm. They used to adapt amoeba like movement. In unfavorable conditions, they
form into mycelium, which in turn produces spores on aerial hyphae. When it returns to
the favorable conditions, spores germinate and undergo rapid cell division to form new
plasmodium.

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REFERENCES:

Text Books:

1. Jeffery C. Pommerville. Alcamo’s Fundamentals of Microbiology (Tenth Edition).

Jones and Bartlett Student edition.

2. Gerard J. Tortora, Berdell R. Funke, Christine L. Case. Pearson - Microbiology: An

Introduction. Benjamin Cummings.

Reference Books:

1. Lansing M. Prescott, John P. Harley and Donald A. Klein. Microbiology. Mc Graw

Hill companies.

2. Biology, Raven and Jhonson, 6th edition (2001)

3. Microbiology, Pelczar. M.J , Chan E.C.S, Kreig N.R, 5th edition (2007)

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