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Mutagenesis
It is a process by which the genetic information of
an organism is changed in a stable manner, resulting in
a mutation.
It may occur spontaneously in nature, or as a result of
exposure to mutagens.
It can also be achieved experimentally using
laboratory procedures
DNA may be modified, either naturally or artificially, by a number
of physical, chemical and biological agents, resulting in mutations
In 1927, Hermann Muller first demonstrated that mutation
with observable changes in the chromosomes can be caused
by irradiating fruit flies with X-ray.
HISTORY :
Lewis Stadler also showed the mutational effect of X-ray on
barley in 1928, and ultraviolet (UV) radiation on maize in 1936.
Types of mutagenesis
1. Directed mutagenesis
Directed mutagenesis can be done using :
Directed mutagenesis may define is a change in the nucleic acid
sequence (or genetic material) of an organism at a specific
predetermine Site. directed mutagenesis is incorporation of a
desired amino acid (of one's choice) in place of a specific amino acid
in a protein or a polypeptide.
Site-directed mutagenesis is the technique for generating amino
acid coding changes in the DNA (gene). By this approach specific
(site-directed) change (mutagenesis) can be made in the base (or
bases) of the gene to produce a desired enzyme.
2. Site-directed mutagenesis
• In molecular biology and genetics, mutations are
changes in a genomic sequence.
• Mutations are caused by radiation, viruses, transposes and
mutagenic chemicals, as well as errors that occur during
meiosis or DNA replication.
• Site-directed mutagenesis, also called site-specific
mutagenesis or oligonucleotide directed mutagenesis, is a
molecular biology technique often used in bio molecular
engineering in which a mutation is created at a defined site
in a DNA molecule.
3. PCR mutagenesis
4. Insertional mutagenesis
5. Signature tagged mutagenesis
6. Transposon mutagenesis
Role of plant Harmon in
mutagenesis
 The experiments are designed to measure the
effects of
- Abscisic acid (ABA)
- Auxin
 Samples are
-Wild-types
- mutated at rop genes
Maize rop mutants
 There are nine rop genes in maize
 used samples with mutation of the rop 2, rop
6 and rop 7 genes.
 How do you get the mutation?
Experiment set up
 Germinate seedlings in a water soaked paper
towel
 Measure root length
before treatment
 Add auxin or ABA
 Measure after 24 and 48hrs
 Average the length
Auxin – rop 6/7
Day Two Comparison
0
20
40
60
80
100
120
0 1 2 3 4
%ofrootgrowh
Wildtype grow th
Mutant grow th
Day One Comparison
0
20
40
60
80
100
120
0 1 2 3 4
%ofrootgrowth
Wildtype grow th
Mutant grow th
Mutant Effect !!
rop 6/7 Double
mutant
SLOWER root
growth
Auxin rop 2
Day One Compaison
0
20
40
60
80
100
120
0 1 2 3 4
%ofrootgrowth
Wildtype grow th
Mutant grow th
Day Two comparison
0
20
40
60
80
100
120
0 1 2 3 4
%ofrootgrowth
Wildtype growth
Mutant growth
No Mutant Effect !!!
rop 2 mutant
SAME as wild-type
ABA rop 2
Day One Comparison
0
20
40
60
80
100
120
0 1 2 3 4
%ofrootgrowth
Wildtype grow th
Mutant grow th
Day Two Comparison
0
20
40
60
80
100
120
0 1 2 3 4
%ofrootgrowth
Wildtype grow th
Mutant grow th
Mutant Effect !!
rop 2 mutant
SLOWER root growth
ABA- rop 6/7
Day One Comparison
0
20
40
60
80
100
120
0 1 2 3 4
%ofrootgrowth
Wild-type growth
Mutant growth
Day Two Comparison
0
20
40
60
80
100
120
0 1 2 3 4
%rootgrowth
Wild-type grow th
Mutant grow th
No Mutant Effect !!!
rop 6/7 mutant
SAME as wild-type
Summary of Results
 ABA,Auxin
 Auxin rop 6/7 Supports hypothesis
Auxin rop 2 No effect
 ABA rop 2 Supports hypothesis
 ABA rop 6/7 No effect
Root growth
Rop
Hormonal Factors of Mutagenesis and
Anti-Mutagenesis Regulation.
Case study 1
Mekhtizadeh Emin RZA
292 92
The genetic activity of phytohormones (Phs) was studied. The investigation was
conducted with 4 plants (Allium fistulosum, Lycopersicon esculentum,
Triticum aestivum, and Gossypium hirsutum). The action of Phs on plant genetic
processes was evaluated by chromosome aberration testing. It was observed that
the exogenic Phs exerted a stabilizing effect on spontaneous plant mutagenesis.
The use of Phs before and after the action of mutagenic factors contributed to an
increase in the anti-mutational or reparative function of the plants´ genes.
It was observed that the Phs exhibited gene protective or reparative activity.
The absence of clearly expressed dependence of these effects on Ph dose was
recorded.
Turkish Journal of Biology 2009Vol. 33 No. 1 pp. 15-20
Conclusion :
Beale, G. (1993). "The Discovery of Mustard Gas Mutagenesis by
Auerbach and Robson
in 1941". Genetics 134 (2): 393–399.
Crow, J. F.; Abrahamson, S. (1997). "SeventyYears Ago: Mutation
Becomes Experimental". Genetics 147 (4): 1491–1496
https://en.wikipedia.org/wiki/Mutagenesis
http://www.cabdirect.org/search.htm
dergipark.ulakbim.gov.tr/tbtkbiology/article/view
Reference :
Harmonal mutagenesis in plant

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Harmonal mutagenesis in plant

  • 1.
  • 2. Mutagenesis It is a process by which the genetic information of an organism is changed in a stable manner, resulting in a mutation. It may occur spontaneously in nature, or as a result of exposure to mutagens. It can also be achieved experimentally using laboratory procedures
  • 3. DNA may be modified, either naturally or artificially, by a number of physical, chemical and biological agents, resulting in mutations In 1927, Hermann Muller first demonstrated that mutation with observable changes in the chromosomes can be caused by irradiating fruit flies with X-ray. HISTORY : Lewis Stadler also showed the mutational effect of X-ray on barley in 1928, and ultraviolet (UV) radiation on maize in 1936.
  • 4. Types of mutagenesis 1. Directed mutagenesis Directed mutagenesis can be done using : Directed mutagenesis may define is a change in the nucleic acid sequence (or genetic material) of an organism at a specific predetermine Site. directed mutagenesis is incorporation of a desired amino acid (of one's choice) in place of a specific amino acid in a protein or a polypeptide. Site-directed mutagenesis is the technique for generating amino acid coding changes in the DNA (gene). By this approach specific (site-directed) change (mutagenesis) can be made in the base (or bases) of the gene to produce a desired enzyme.
  • 5. 2. Site-directed mutagenesis • In molecular biology and genetics, mutations are changes in a genomic sequence. • Mutations are caused by radiation, viruses, transposes and mutagenic chemicals, as well as errors that occur during meiosis or DNA replication. • Site-directed mutagenesis, also called site-specific mutagenesis or oligonucleotide directed mutagenesis, is a molecular biology technique often used in bio molecular engineering in which a mutation is created at a defined site in a DNA molecule.
  • 6. 3. PCR mutagenesis 4. Insertional mutagenesis 5. Signature tagged mutagenesis 6. Transposon mutagenesis
  • 7. Role of plant Harmon in mutagenesis  The experiments are designed to measure the effects of - Abscisic acid (ABA) - Auxin  Samples are -Wild-types - mutated at rop genes
  • 8. Maize rop mutants  There are nine rop genes in maize  used samples with mutation of the rop 2, rop 6 and rop 7 genes.  How do you get the mutation?
  • 9. Experiment set up  Germinate seedlings in a water soaked paper towel  Measure root length before treatment  Add auxin or ABA  Measure after 24 and 48hrs  Average the length
  • 10. Auxin – rop 6/7 Day Two Comparison 0 20 40 60 80 100 120 0 1 2 3 4 %ofrootgrowh Wildtype grow th Mutant grow th Day One Comparison 0 20 40 60 80 100 120 0 1 2 3 4 %ofrootgrowth Wildtype grow th Mutant grow th Mutant Effect !! rop 6/7 Double mutant SLOWER root growth
  • 11. Auxin rop 2 Day One Compaison 0 20 40 60 80 100 120 0 1 2 3 4 %ofrootgrowth Wildtype grow th Mutant grow th Day Two comparison 0 20 40 60 80 100 120 0 1 2 3 4 %ofrootgrowth Wildtype growth Mutant growth No Mutant Effect !!! rop 2 mutant SAME as wild-type
  • 12. ABA rop 2 Day One Comparison 0 20 40 60 80 100 120 0 1 2 3 4 %ofrootgrowth Wildtype grow th Mutant grow th Day Two Comparison 0 20 40 60 80 100 120 0 1 2 3 4 %ofrootgrowth Wildtype grow th Mutant grow th Mutant Effect !! rop 2 mutant SLOWER root growth
  • 13. ABA- rop 6/7 Day One Comparison 0 20 40 60 80 100 120 0 1 2 3 4 %ofrootgrowth Wild-type growth Mutant growth Day Two Comparison 0 20 40 60 80 100 120 0 1 2 3 4 %rootgrowth Wild-type grow th Mutant grow th No Mutant Effect !!! rop 6/7 mutant SAME as wild-type
  • 14. Summary of Results  ABA,Auxin  Auxin rop 6/7 Supports hypothesis Auxin rop 2 No effect  ABA rop 2 Supports hypothesis  ABA rop 6/7 No effect Root growth Rop
  • 15. Hormonal Factors of Mutagenesis and Anti-Mutagenesis Regulation. Case study 1 Mekhtizadeh Emin RZA 292 92 The genetic activity of phytohormones (Phs) was studied. The investigation was conducted with 4 plants (Allium fistulosum, Lycopersicon esculentum, Triticum aestivum, and Gossypium hirsutum). The action of Phs on plant genetic processes was evaluated by chromosome aberration testing. It was observed that the exogenic Phs exerted a stabilizing effect on spontaneous plant mutagenesis. The use of Phs before and after the action of mutagenic factors contributed to an increase in the anti-mutational or reparative function of the plants´ genes. It was observed that the Phs exhibited gene protective or reparative activity. The absence of clearly expressed dependence of these effects on Ph dose was recorded. Turkish Journal of Biology 2009Vol. 33 No. 1 pp. 15-20 Conclusion :
  • 16. Beale, G. (1993). "The Discovery of Mustard Gas Mutagenesis by Auerbach and Robson in 1941". Genetics 134 (2): 393–399. Crow, J. F.; Abrahamson, S. (1997). "SeventyYears Ago: Mutation Becomes Experimental". Genetics 147 (4): 1491–1496 https://en.wikipedia.org/wiki/Mutagenesis http://www.cabdirect.org/search.htm dergipark.ulakbim.gov.tr/tbtkbiology/article/view Reference :