BACTERIAL GENETICS

& REPRODUCTION
DR. M. ASHRAF, Ph.D.
(MICROBIOLOGY & MOLECULAR
GENETICS)
NIAB, PIEAS, PAEC, FAISALABAD
 BACTERIAL REPRODUCTION
Bacteria
Simplest, the smallest, most successful microorganisms
First discovered by Anton Leeuwenhoek (1676)
Placed in kingdom Monera -in the five kingdom classification
Modes of Reproduction
Vegetative
Binary Fission
Budding
Fragmentation

Asexual
Formation of Conidiospores or sporangiospores

Sexual
Conjugation
Transformation
Transduction

Dr.M.Ashraf,PS,NIAB,PIEAS,PAEC 2
 VEGETATIVE
Binary Fission (A, B, C)
Transverse cell wall binary fission
Most common in bacteria
During the process
Bacterial chromosomes get attached to the cell membrane
Replicates and separated as the cell enlarges
A cross wall -forms b/w the separating daughter
chromosomes
The cell divides into two daughter cells
Daughter cells soon grow to maturity within 20-30 minutes
Under favorable conditions many bacteria divide once in 20-30
min

Budding (D, E)
Small protuberance (bud) develop at one end
Genome replication follows
copy of the genome gets into the bud
Bud enlarge and develop into new daughter cell

Dr.M.Ashraf,PS,NIAB,PIEAS,PAEC 3
VEGETATIVE REPRODUCTION
Fragmentation (F)
Found in the bacteria with extensive filamentous growth
Filaments fragments into small bacillary or coccoidal cells

OR
Mostly during unfavorable conditions
Bacterial protoplasm undergoes
Compartmentalization and subsequent
fragmentation, forming minute bodies called Gonidia
Under favorable conditions, each gonidium grows to a
new bacterium
During fragmentation the bacterial genome has to
undergo repeated replication so that each fragment
gets a copy of it

Dr.M.Ashraf,PS,NIAB,PIEAS,PAEC 4
 BACTERIAL REPRODUCTION

A Bacillus subtilis
B Streptococcus faecalis

C Prosthecobacter fusiformis

D
Rhodopseudomonas acidophila
E
Hyphomicrobium vulgare
F Nocardia sp.

G Streptomyces sp.
Conidiospores

Dr.M.Ashraf,PS,NIAB,PIEAS,PAEC 5
 ASEXUAL REPRODUCTION
Asexual reproduction takes place by
Endospore formation, Conidia, Zoo spores

A. Endospore
Very resistant to extreme physical conditions and chemicals
During unfavourable conditions
Resting spores formed in some gram positive bacteria
Bacillus and Clostridium
Spores formed within the cells
During spore formation
A part of the protoplast becomes concentrated around the
chromosome
A hard resistant wall is secreted around it
Rest of the bacterial cell degenerates
During favourable conditions
Spore wall gets ruptured
Protoplasmic mass gives rise to a new bacterium
B. Formation of Conidiospores (G)
Genus Streptomyces produces many spores per organism by
cross walls (Septation) at the hyphal tip 6
Dr.M.Ashraf,PS,NIAB,PIEAS,PAEC
 SEPTUM FORMATION
Septum
Dividing plate in the bacteria formed at cell division
Does not begin to form until doubling of the chromosome (Cell division
is triggered by completion of DNA replication)
Steps in Septum Formation
1. Inward growth of the cytoplasm membrane at middle of the cell
At the site a Mesosome attached
Mostly in G+ve
May have role in synthesis of new membrane
2. Inward growth of cell wall to form septum
Ultimately splits to form two cells
.S. faecalis (Figure A1)
i. Septum formation beneath equatorial ridge in the cell wall
ii. New cell wall material synthesized
iii. Septum formed contributes to half of the cell wall of each new cell
iv. Hemispherical shape formed due to plasticity created due to
a) The turgor pressure of the protoplast against newly
synthesized wall
b) A certain amount of reorganization of peptidoglycan
Resulted from breakage of some of the chemical
bonds by hydrolysing enzymes & subsequent
formation of new bonds at different location
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Dr.M.Ashraf,PS,NIAB,PIEAS,PAEC
 SEPTUM FORMATION
B. subtilis
i. Septum formation similar to S. faecalis
ii. No ridge present
iii. Only 15% of the wall derived from septum
iv. Rest I synthesized by cylindrical part of the cell
v. So Bacillus grows by elongation rather than just septum formation
vi. Hemispherical shape formed due to plasticity created due to
a) The turgor pressure of the protoplast against newly
synthesized wall
b) A certain amount of reorganization of peptidoglycan
Resulted from breakage of some of the chemical
bonds by hydrolysing enzymes & subsequent
formation of new bonds at different location
E. coli (G-ve bacteria) Figure A2
i. Formation of a belb or fold of outer membrane at site of septum
ii. Outer membrane does not invaginate until final stages of septum
iii. Cytoplasmic membrane & peptidoglycan layers grows inward in
early stages
iv. A mesosome is present during early stages of septum formation

Dr.M.Ashraf,PS,NIAB,PIEAS,PAEC
SEPTUM FORMATION

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Dr.M.Ashraf,PS,NIAB,PIEAS,PAEC
 SEXUAL REPRODUCTION
Sexual reproduction
occurs in the form of Genetic Recombination
Three main methods of Genetic Recombination
Transformation, Transduction , Conjugation
Conjugation
Transfer of genes b/w bacterial cells in physical contact to each other
First reported by Lederberg and Tatum (1946) in E. coli
Cell to cell union occurs between two bacterial cells
Genetic material (DNA) of one bacterial cell goes to another cell
lengthwise through conjugation tube formed by sex pili.
Transduction
Transfer of genes from one bacterial cell to other by a phage
First reported in Salmonella typhimurium (Zinder and Lederberg,
1952)
Genetic material of one bacterial cell goes to other bacterial cell by
agency of bacterio-phages or phages (viruses, infecting bacteria)
Transformation
Transfer of cell-free or Naked DNA from one cell to another
First studied by Griffith (1928) in Diplococcus pneumonia
Genetic material of one bacterial cell goes into another bacterial cell
by some unknown mechanism
Converts one type of bacterium into another type (non
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capsulated to
capsulated form)
Dr.M.Ashraf,PS,NIAB,PIEAS,PAEC
CONJUGATION

Dr.M.Ashraf,PS,NIAB,PIEAS,PAEC 11
 CONJUGATION
Lederberg & Tatum Expt.
Coculturing of auxotroph of E. coli
Screening on Prototrophic medium
Prototroph
Synthesize their own metabolites without any additional growth factor
Auxotroph
Require addition of specific nutrients to the medium
Polyauxotroph
Mutants with more than one nutritional requirement
Sexual differentiation in E. coli
Male contain Sex factor or F (fertility) factor
Referred as F+
Small circular piece of DNA present in cytoplasm not part of
chromosome
F+ Donor cell F- recipient cell
Cross b/w F- -No recombination
F+ x F- -Recombinants -Converts F- to F+ -the donor cell
F factor
Carries 40 genes to specify & control replication of sex factor and
synthesis of sex pili -the tubules through which DNA passes from
F+ to F-
One or more Pili are produced by each F+ cell
Frequency of f factor transfer 100% recombination 104 105
Transfer of F factor independent of chromosome
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Dr.M.Ashraf,PS,NIAB,PIEAS,PAEC
 CONJUGATION
Hfr strains of E. coli
High recombinant capability
103 times higher than F+ X F- cross
Plasmids
Extra chromosomal genetic elements capable of independently
replicate in the cytoplasm
Auxotroph
Require addition of specific nutrients to the medium
Episomes
Plasmids capable of integrating into chromosome
F+ is an episome it can integrate into chromosome to form Hfr
(High frequency recombination factor)
Bactericinogenic
Plasmids having bacteriocin synthesis factor
Kills the same or related species
Colicin the bacteriocins of E. coli
Pycocins Pseudomonas aeruginosa
R plasmids
Plasmids conferring resistance to number of antibiotics
Infectious resistance Transfer of R plasmid to other non
resistance cells through conjugation

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Dr.M.Ashraf,PS,NIAB,PIEAS,PAEC
 BACTERIAL GENETICS
Replication of DNA
Semi-conservative
2 strands separated and act as template for new DNA synthesis
DNA helix contains one new and an old strand
Bacteria
Always haploid unpaired chromosome
Genetic loci in a single linkage group i.e., single chromosome per
genome
Circular double stranded DNA -double helix for a complete genome
No free ends
MW approx. 2.5 x 109 daltons (dalton = mass of 1H1)
4 x 106 base pairs
Chromosome supercoiled
1250 um (1.25 mm) if extended linearly
Several hundred times larger than a typical bacterial cell
Three modes of replication
Theta () Mode
Sigma () Mode
Linear Mode

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Dr.M.Ashraf,PS,NIAB,PIEAS,PAEC
 BACTERIAL GENETICS
Theta () Mode of Replication

Replication initiated at certain point the Origin

Specific for bacterial strain
Replication in two directions
Forming a bubble
Bubble increases in size as the replication goes on
Mode is () (Greek letter) due to resemblance
Circular parental chromosome replicated in 2 daughter
circular chromosome
One strand of DNA conserved other newly synthesized

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Dr.M.Ashraf,PS,NIAB,PIEAS,PAEC
 BACTERIAL GENETICS
Sigma () Mode of Replication
Replication by cleavage of phosphodiester bond in one strand
Produce nick with 3-OH and 5 -PO4 ends on the strand
Complimentary circular strand act as template for synthesis of
new strand
Covalently linked to 3 -OH end of the nicked parent
strand
As the strand grows the 5 -PO4 end displaced to form a
Tail on the circular
As the replication goes on -a circular parent DNA molecule
converts to 2 daughter molecules
a. One linear
b. One circular
The mode is Sigma as the structure has () conformation
Carried out by some bacteriophages e.g., and X174
-whole progeny viral DNA
-linear by bacteria involved in sexual conjugation

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Dr.M.Ashraf,PS,NIAB,PIEAS,PAEC
 BACTERIAL GENETICS
Linear Mode of Replication
Some viruses & Eukaryotes have linear DNA replication
Replication starts at specific site by formation of bubbles
Small molecules have one point of initiation
Eukaryotes hundreds of initiation points
Replication bubble grows in size as DNA replication goes on
Two linear double helix daughter molecules form upon
completion of replication
The daughter molecules are each with a parental and a newly
synthesized strand
Prokaryotes one origin of replication
Replication is either unidirectional or bidirectional

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Dr.M.Ashraf,PS,NIAB,PIEAS,PAEC
 TRANSDUCTION
Transfer of bacterial DNA through Phages
Generalized Transduction
Each fragment of degraded chromosome has a chance to be packed
in the phage head
Specialized Transduction
Specific restricted part of bacterial chromosome especially
the genes adjacent to prophage are transferred by phage
Characteristics of temperate bacteriophages e.g.,

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Dr.M.Ashraf,PS,NIAB,PIEAS,PAEC