Mutation breeding is a technique that uses mutagens like radiation and chemicals to induce mutations in plant genomes to create genetic variability for plant breeding purposes. The key steps in mutation breeding are selecting plant material, treating it with mutagens, growing the treated material to identify desirable mutants. Notable mutant varieties released in India include MA-9 cotton and several disease resistant chickpea and mung bean varieties. While mutation breeding can create useful variation, it also has limitations like low mutation rates and risk of undesirable side effects. Overall, mutation breeding remains an important tool for plant breeding.

1. Mutation
Breeding

2. Mutation Breeding
Presented by :
Chaudhary Ankit R.
Submitted to :
Dr. N. V. Soni
Department of Genetics & Plant Breeding
C .P . College of Agriculture
Sardarkrushinagar, Dantiwada
Reg. No. : 04-AGRMA-01571-2017

3. INTRODUCTION
 Plant Breeding:
• It can be defined as an art and science of improving
the genetic make up of plants in relation to
economic use.
 Types of Breeding methods
1. General Breeding methods
2. Special Breeding methods

4. SPECIAL BREEDING TECHNIQUES
 There several types of Special Breeding techniques.
 Mutation Breeding
 Haploid and Aneuploidy
 Polyploidy
 Distance Hybridization
 Tissue culture
 Somatic Hybridization
 Transgenic Breeding

5. MUTATION
BREEDING

6. • Mutation was first discovered by
Wright in 1791.
• The term mutation was coined by
de Hugo de Vries in 1900.
• Muller – Discover the mutate action
of X-rays on Drosophila and
Stadler discovered the effect of X-
rays in barley in 1927.
• Mutation breeding programme was
started in Sweden, USSR and
Germany in 1927 after the
discovery by muller.
History

7. MUTATION BREEDING
 Mutation: Sudden heritable change in a
characteristic of an organism not due to segregation
and recombination.
 It is commonly used in self pollinated and asexually
propagated species.
 Mutation range : 10-5 to 10-6
 Mutation do occurs by two way
(I) In Nature or Spontaneous mutation
(II) Artificial mutation or Induced Mutation

8.  Mutations result from
 Unrepaired damage to DNA or to RNA genomes
(typically caused by radiation or chemical mutagens),
 The insertion or deletion of segments of DNA
Characteristics of Mutation
 Mutations are generally recessive.
 Mutations are generally harmful to the organisms. Most of
the mutations have deleterious effect, but a small
proportion of them are beneficial. (0.1 %)
 Mutation are random.
 Mutation are recurrent.
 Induce mutation commonly show pleiotropy often due to
mutation in closely linked genes.

9. CLASSIFICATION OF MUTATION
By effect on structure of chromosome (Chromosomal Mutation)
 Base Insertions
 Deletions
 Duplications
 Inversions
 Translocations

10. Insertions
Insertion of one or more bases into a DNA molecule.

11. Deletions
Loss one or more nucleotides from the DNA

12. Duplications
Doubling of chromosomal segments

13. Inversions
changing orientation of a DNA segment.

14. • Translocations
A change in location of a chromosome segment is a
translocation.

16. CLASSIFICATION OF MUTATION
 Based on Sourse:
-Spontaneous mutation
-Induced mutation
 Based on Direction:
-Forward mutation
-Reverse mutation
 Based on Tissue:
-Somatic mutation
-Germinal mutation

17.  Based on Survival:
-Lethal mutation (100% mortality)
-Sub-lethal mutation (>50% mortality)
-Sub-vital mutation (< 50% mortality)
-Vital mutation (all mutants survive)
 Based on Site:
-Nuclear mutation
-Cytoplasmic mutation

18.  Based on Character:
-Morphological mutation
-Biochemical mutation
 Based on Visibility:
-Macro mutation
-Micro mutation

19. OTHER TYPES OF MUTATION
 By effect on fitness
 harmful mutation
 beneficial mutation
 deleterious mutation
 advantageous mutation
 By effect on function
 Loss-of-function mutations
 Gain-of-function mutations
 Reversion / Back mutation
 Lethal mutations

20. By impact on protein sequence
(Gene mutation)
• Point mutations often caused by chemicals or malfunction of
DNA replication, exchange a single nucleotide for another.
• Frameshift mutation: is a mutation caused
by insertion or deletion of a number of nucleotides.
• Silent mutations: which code for the same (or a sufficiently
similar) amino acid.
• Missense mutations: which code for a different amino acid.
• Nonsense mutations: which code for a stop codon.
• Neutral mutation is a mutation that occurs in an amino acid
codon which results in the use of a different, but chemically
similar, amino acid.

22. Neutral
New amino acid has the
chemical properties as the
old amino acid
Change in a base pair
does not result in a change
in the sequence of amino
acids in a protein

23. MUTAGENS
 Mutagen:- Physical or chemical agent which enhance the
frequency of mutation.
 Physical mutagens:
1. X-rays :- S.I., penetrating, non particulate
 Induce mutation by forming free radicles and ions, cause
addition, deletion, transition, transversion.
2. Gamma rays:- S.I., very penetrating, non particulate.
 Induced mutation by ejecting atoms from tissues, cause
addition, deletion, transition, transversion.

24. 3. Alpha particles:- D.I., particulate, less penetrating,
positively charged
 Ionization & excitation, cause chromosomal & gene
mutation
4. Beta rays:- S.I., negatively charged, more penetrating
 Act as alpha particles.
5. Fast & Thermal neutrons:- D.I., neutral particles, highly
penetrating
 Cause chromosomal breakage & gene mutation.

25.  6. U.V rays:- non – ionizing, low penetrating
 Act as fast & thermal neutrons & produce dimers of
thymine, uracil and sometimes guanine

26. MODE OF ACTION OF UV
RADIATION
UV radiation promotes the formation of covalent
bonds between adjacent thymine residue in a
DNA strand creating a cyclobutyl ring.
They form abnormal chemical bond between
adjacent pyrimidine molecule (mainly thymine) in
the same strand of the double helix.

27.  They have two main chemical effects on pyrimidines.
 The first effect is the additions of a water molecule
which weakens the H bonding and permits separation
of DNA strands.
 The second effect is to joining of pyrimidines to make a
pyrimidines dimer. Thus dimerization can produce TT,
CC, UU and mixed pyrimidine dimers like CT.
 Dimerization interferes with DNA and RNA synthesis.
Interstrand dimers cross link nucleic acid chains,
inhibiting strand separation and distribution.

29.  When an X-ray photon collides with an atom, the
atom may absorb the energy of the photon and
boost an electron to a higher orbital level.
 Depending on the structure of the atom and the
energy of the X-ray photon, it may knock an
electron from the atom altogether, causing the atom
to ionize.
 Generally, a larger atom is more likely to absorb an
X-ray photon in this way, since larger atoms have
greater energy differences between orbital
electrons.
Mode of action of X-ray

30.  Ionizing radiation penetrates tissue, colliding with
molecules and knocking electrons out of orbits &
creating ions.
The ion can result in the breakage of covalent
bonds, including those in the sugar phosphate
back bone of DNA.
 Ionizing radiation is the leading causes of gross
mutation in humans.
 High dosages of ionizing radiation kill cells so
use in treating some form of cancer.

31.  Chemical mutagens:
1. Alkaylating agents:
 Ethyl methane sulphonate : AT – GC Transition
 Methyl methane sulphonate: Transition
 Ethyl ethane sulphonate: GC- AT Transition
 Ethylene Imines: Transition
2. Base Analogues:
 5-Bromo uracil : AT - GC Transition
 2-Amino purine : AT - GC Transition

32. 3. Acridine Dyes:
 Acriflavin, Proflavin- Cause addition, deletion and
frameshift
4. Others:
Nitrous Acid : AT – GC Transition
Hydroxylamine : GC – AT Transition
Sodium Azide : Transition

33. MODE OF ACTION OF ALKYLATING AGENT
They transform the alkyl groups to the phosphate
residues of DNA produce triesters.
They are readily transforms to 6-oxygen and 7-
nitrogen of DNA bases.
During the repair of alkylated bases, errors may
occasionally occur producing transitions,
transversions or even frame-shift mutations.
Some alkylating agents, especially those having two
or more active alkyl groups, cross-link DNA strands
and/or DNA molecules; such cross-link prevents DNA
replication and induce chromosomes breakage
resulting in structural aberrations.

34. It often result in depurination , causing a gap
in DNA strand and lead to breakage in the
strand.
Alkylating agents produce a variety of genetic
effect. viz., transitions, transversions,
frameshift mutations, chromosome breakage
etc.

35. MODE OF ACTION: 5-BROMOURACIL
 Br in 5th carbon position increase the frequency of
tautomeric shift.
 Thus keto form will change into enol form by shifting
H atom from its structure.
 The new form is called tautomer.

36. So here, the 5-Bu incorporated into DNA in place
of thymine or cytosine.
During replication it will pair with guanine or
adenine respectively.
Then in next round of replication it will yield their
normal base pair of that
Thus it will cause transitions of A:T to G:C and
G:C to A:T

38. MODE OF ACTION OF ACRIDINE DYES
Acridine dyes get inserted between two base
pair of DNA and lead to addition or deletion of
single or few base pair when DNA replicates.
Thus, they cause frame shift mutation
Acridine dyes bind directly to the DNA by
using their positive charge.
 Acridine dyes can insert or delete only one base
pair in DNA, and thus result in frameshift
mutations

39. PROCEDURE FOR MUTATION
BREEDING
 The mutation breeding process consist of four
important steps.
 Choice of Material
 Part of the plant to be treated
 Mutagen treatment
 Handling of treated material.
India has released 259 mutant variety and 2nd in
world after china (605)

40. FIG. 1 A GENERALIZED SCEME OF MUTATION BREEDING
First year M1 i
ii
Mutagen-treated seeds space planted
Seeds from individual plants harvested
separately
Second year M2 i
ii
Individual plant progenies grown
Fertile, vigorous, normal looking plants
harvested separately
Third Year M3 i
ii
Individual plant progenies from the selected
plants grown
Superior plant selected from superior
progenies showing segregation
Fourth year M4 i
ii
iii
Individual plant progenies from the selected
plants grown
Superior homogeneous lines harvested in bulk
Segregating lines usually rejected
Fifth year M5 i
ii
Preliminary yield trail with a suitable check
Superior lines selected
Sixth to
eight year
M6
to
M8
i
ii
Replicated yield trial with at several locations
Outstanding line released as a new variety
Nine year M9
I Seed increase for distribution among farmers
─ ─ ─ ─ ─
─ ─ ─ ─ ─
─ ─ ─ ─ ─
• • • • •
• • • • •
• • • • •
• • • •
•

41. IMPORTANCE OF MUTATION BREEDING
 When a high yielding variety has oligogenic defect such as
susceptibility to disease, mutation breeding is the best
source of a line for breeder.
 When there is tight linkage between desirable and
undesirable characters mutation breeding is the best way of
overcoming such problems.
 When in fruit cops, the improvement has to be made
without change in taste and color of fruit it can be
achieved through mutation breeding.
 Where generation cycle is very long, such as fruit trees and
forest trees, mutation breeding is the only short cut
method.

42.  From 1930 to 2014 more than 3200 mutagenic plant
varieties were released that have been derived either as
direct mutants (70%) or from their progeny (30%).
 However, although the FAO/IAEA reported in 2014 that
over 1,000 mutant varieties of major staple crops were
being grown worldwide. It is unclear how many of these
varieties are currently used in agriculture or horticulture
around the world.

43.  Co-4, Pant Mung-2, and TAP mung bean mutants
 MA-9 cotton - the world's first mutant cotton, released in
1948 (X-ray radiation; drought tolerance, high yielding)
 PNR-381 (Rice)
 Pusa 408 (Ajay), Pusa 413 (Atul), Pusa 417 (Girnar), and
Pusa 547 chickpea mutants (resistant to Ascochyta blight and
wilt diseases, and have high yields)
 Sharbati Sonora (wheat)
 Tau-1, MUM 2, BM 4, LGG 407, LGG 450, Co-4, Dhauli
(TT9E) and Pant moong-1(Yellow mosaic virus resistance)
 TG24 and TG47 (groundnut mutants)
Notable Mutagen Varieties released by India

44.  As of 2011 the percentage of all mutagenic varieties released
globally, by country, were:
 (25.2%): People’s Republic of China
 (15.0%): Japan
 (11.5%): India
 (6.7%): Russia
 (5.5%): Netherlands
 (5.3%): Germany
 (4.3%): United states
 (2.4%): Bulgaria
 (1.7%): Vietnam
 (1.4%): Bangladesh

45. APPLICATIONS
 Induction of desirable mutant allele, which is not present in
germplasm
 Improving specific character of variety without change in
genetic make up
 Used to improve quantitative characters
 Induction of male sterility
 Production of haploid by irradiating pollens
 Overcoming self incompatibility

46. LIMITATIONS
 Low frequency
 Need screen large number of population
 Pleiotropic effects.
 Identification of micro mutation is very difficult
 Desirable mutations are generally associated with undesirable
side effects.
 Most of the mutations are recessive.
 Most of the mutations are deleterious and undesirable.

47.  Mutation induction has been proved to be a good
means of obtaining useful genetic variability.
 Mutant can be directly released as a variety.
 Induced mutants utilized in recombination breeding
evolved several distinct lines, with improved agronomic
traits.
 Gamma ray and EMS mutagens were found to be
effective at 150-300 Gy and 0.5%, respectively to
induce variation.
 Mutation induced earliness, disease resistance,
drought tolerance, change in physiological and
biochemical characters which are of interest to evolve
superior genotypes for breeding purpose.

48. MUTATION BREEDING
48