Master’s Seminar

on
Mutation breeding in fruit crops

Presented by:
Abdullah Zaid
Id. No. 2053
Department of Fruit Science
College of Horticulture
Banda University of Agriculture & Technology, Banda-210001
 Mutation
• Term mutation was coined by Hugo De Varies in the year 1900 while
studying evening prime rose
• He put forward mutation as “sudden heritable changes in a character of an
organism”
• Mutation is an alteration of the nucleotide sequence of the genome of an
organism
• Individual showing these changes are known as mutant
• Factors which causes mutation are known as mutagens
• Mutation may be harmful, beneficial or neutral
 History
 English farmer Seth Wright recorded case of mutation first time
in 1791 in male lamb with unusual short legs
 Term mutation was coined by Hugo De Varies in the year 1900
 Systematic study of mutation was started in 1910 when Morgan
genetically analyzed white eye mutant of drosophila
 H.J. Muller induced mutation in drosophila by using X-ray in
1927, he was awarded with Nobel prize in 1946
 Stadler discovered the effect of gamma rays in barley
 Mutation breeding program was started in Sweden, USSR and
Germany in 1927 after the discovery by Muller
 How do Mutations occur?
 Sometimes during
replication, the cell makes a
mistake and adds the wrong
base
 When the cell replicates its
DNA again, the two strands
that are produced are no
longer exactly the same
 This usually will cause the
new cell to die, but
sometimes it can cause the
cell to divide when it is not
supposed to  cancer
 Types of Mutation
 Spontaneous mutation - They are naturally occurring. Spontaneously
arising mutations are very rare and random events in terms of the
time of their occurrence and the gene in which they occur.
 Induced mutation- mutation may be artificially induced by treatment
with certain physical and chemical agent.
 to induce mutations in crops, planting materials are exposed to
physical and chemical mutagenic agents. Mutagenesis can be
performed with all types of planting materials, Multiple forms of
plant propagules, such as bulbs, tubers, corms, shoot tip, leaf, ovules,
protoplasts and rhizomes.
 Mutagens
It is a physical or chemical Agent that induces the frequency
of mutation , are known as mutagens

Type of mutagens

Physical mutagens:-
(1) Ionizing radiation
(a) Particulate radiation- Alpha rays, Beta rays, Thermal neutron
(b) Non-particulate radiation- X- ray and Y- ray
(2) Non-ionizing radiation – Ultra violet radiation
 Chemical Mutagens

A. Alkylating agents – Ethyl Methane Sulphonate(EMS) Methyl

Methane Sulphonate(MMS), Sulphur Mustard, Nitrogen
Mustard
B. Acridine Dyes – Acriflavine, Proflavine, Acridine Orange,
Acridine Yellow.

C. Base analogues- 5- Bromouracil , 5-Chlorouracil

D. Other- Nitrous Acid, Hydroxyl Amine.

 Physical Mutagens
 In the past 80 years, physical mutagens, mostly ionizing radiations, have
been used widely for inducing hereditary aberrations and more than 70%
of mutant varieties were developed using physical mutagenesis (Mba et
al., 2012). Radiation is defined as energy travelling through a distance in
the form of waves or particles. These are relatively high energy levels of
electromagnetic (EM) spectrum that are capable of dislodging electrons
from the nuclear orbits of the atoms that they impact upon. These
ionizing components of the electromagnetic include cosmic, gamma and
X-rays. X-rays were the first to be used to induce mutations and gamma
radiation from radioactive cobalt ( 60Co) is widely used. It has high
penetrating potential and is hazardous
 Sources of Radiation
MUTAGEN SOURCES CHARACTERISTICS HAZARDS

Gamma rays Radioisotopes and Creates Dangerous and very

nuclear reaction electromagnetic penetrating-
radiation that
penerates into tissues

X-rays X-ray machine Penerates tissues from Dangerous and

few milimetres penetrating

Alpha particles Radioisotopes Helium nucleus Very dangerous

capable of heavy
ionization
Beta particles Radioisotopes or Shallowly penetrating May be dangerous
Acclerators
Gamma field at the Institute of
Radiation Breeding in Japan
Examples of Radio sensitivity of
tissue in fruit crops
Species Mutagen Planting material
Apple Gamma rays Leaves
Japanese Plum Gamma rays Shoot
Strawberry Gamma rays Shoot clump
Kiwi Gamma rays Leaves
Banana Gamma rays Shoot tip
Pineapple Gamma rays Crown
Pear Gamma rays in vitro shoots
 Chemical Mutagens
 Chemical mutagens were found to be highly effective in inducing true gene
mutations and the specificity of action could be investigated through analysis
of their reaction with different DNA base. There are different chemical
mutagens used to induce mutation in fruit crops namely alkylating agents
most useful for mutation Ethyl Methane Sulphonate (EMS), Diethyl Sulphate
(DES), Ethyleneimine (EI), . As compared with physical mutagens, chemicals
may give rise to relatively more gene mutations rather than to chromosomal
changes.
 EMS was used for inducing mutations in Banana by treating shoot tips and
then regenerating adventitious buds
 Ems is very efficient in creating of Somaclonal variation in crop plants such
as Banana and Grapes
Chemical Mutagens
Mutagen Example Mode of action
Alkylating agents 1-methyl-1-nitrosourea React with bases and
(MNU), 1-ethyl-1- add methyl or ethyl
nitrosourea(ENU), Methyl groups and, alkylated
methanesulphonate(MM base may degrade to
S), yield an abasic site,
Ethylmethanesulphonate which is mutagenic and
(EMS), recombinogenic, or
mispair to result in
mutations upon DNA
replication.
Azide Sodium azide Same as alkylating
agents
Hydroxylamine Hydroxylamine Same as alkylating
agents
Antibiotics Actinomycin D, Chromosomal
Mitomycin C, Azaserine, aberrations also reported
Sreptonigrin to cause cytoplasmic
male sterility
 Chimera
 A plant or plant part composed of genetically different layers
 Vegetative mutations are expressed in the forms of chimeras
 Periclinal chimera ( whole of inner and outer layer)
 Sectorial chimera (part of inner/outer layer)
 The most common example is a variegated plant where different regions of
leaves are yellow or white due to lack of chlorophyll synthesis these are
chlorophyll mutants
 Partial reversion of red
Sectorial chimera
delicious
 Merit
 The generation cycle is very long such as plantation crops, fruit
trees there mutation breeding is the shortcut way for genetic
improvement
 Induction of desirable mutant alleles which may not be present
in the Germplasm available to the breeder
 In improving specific characteristics of well adapted high
yielding variety
 Mutagenesis has been successfully used to improve various
quantitative characters such as yield and colors
 Mutation breeding is Simple , quick and most effective to
induce a new character in vegetatively propagated cops
Mutations breeding is applied
when:-
 When a desirable variety has Oligogenic genetic defect
 Desirable variability exhaust in cultivated species and
germplasm
 There is tight linkage between desirable and undesirable traits
 Only one or two characters are to be improved in fruit crops
without changing its taste
Spontaneous Mutants in fruit crops
crop Original variety Mutant cultivar traits

Mango Rosado de Lea Rosica Haden Reguler bearer and

Davis haven larger fruit size
Banana Highgate Motta Gros Michael Semi dwarf
Poovan Poovan
Grape fruit Foster Hudson Deep red flesh
colour
Pear clapp‟s Favourite Starkrimson Spotting of colours

Mandarin Owari Pongan Clausellina Pongan

86-1
Navel Orange Bahia Washington Baianinha
Navelina,
Navelate,Marrs,
Leng, Autumn
Gold, Powell
Summer, Winter
Rosica Gross Michael Hudson

Starkrimson Clausellina Pongan Autumn Gold

Achievements through mutation
breeding
species Year Mutagen Cultivar released traits
Apple 1986 Gamma rays Golden Haidegn Large fruit size
1970 Gamma rays Lysgolden Rust resistance
1972 Gamma rays Courtavel Shortness
1985 gamma rays Senbatsu-Fuji-2- Fruit colour
Kei
Peach 1986 Gamma rays Mangnif 135 Fruit size
1981 Gamma rays Plovdiv 6 yield
Banana 1993 Gamma rays Novaria Earliness
1985 Gamma rays, in Klue Hom Thong Bunch size
vitro KU1

Orange/Mandari 1987 X-rays Valencia 2 INTA Fruit set, quality

n 1987 X-rays Eureka 22 INTA Fruit set, quality
Papaya 1986 Gamma rays Pusa nanha dwarfness
Pear 1996 Gamma rays Kotobuki shinsui Disease
resistance
Cont.
species Year Mutagen Cultivar traits
released
Apricot 1979 ThN early Blenheim earliness
Grape 1986 Gamma rays fikreti earliness
Almond 1987 Gamma rays supernova Fruit size
Plum 1988 Gamma rays spurdente- earliness
Ferco
Sour Cherry 1977 X-rays Plodorodnaya Fruit set
1979 Gamma rays Michurina Dwarfness
1979 Gamma ray Plodorodnaya Dwarfness
Orlovskoi
Karlik
Samorodka
Sweet Cherry 1983 Gamma rays Burlat C1 Compact
1992 X-rays Ferrovia spur growth
1974 X-rays Compact Stella Dwarfness
35B11 Compact
growth
Pusa Nanha Star Ruby Chaubhatia
Agrim

Belrene Red Nigra Lapins

DNA Markers for assessment of
genetic diversity
fruit marker
Apple AFLP and RAPD
Avocado Mini satellite DNA
Banana RAPD
Citrus RFLP
Pistachio Mini satellite marker
Pear SSR and AFLP
Cashew RAPD and ISSR
Data

Country wise registered mutant

Fruit categories varieties
 Limitations

 Frequency of desirable mutants is very low

 Desirable mutations are commonly associated with undesirable
side effects
 Problem in registration of mutant variety
 Most of the mutations are recessive
 Many mutations are non heritable
 Future Prospects

More research into directed mutagenesis in vegetative fruit crops is

needed. In fruit crops, the effects of combined mutagen treatment
and recurring mutagenesis must be explored. The use of markers to
detect mutations at an early stage is also required. More research is
needed on in vitro mutation breeding and CRISPR/Cas-9
approaches, especially for fruit crops.
 Conclusion

 Mutation is important breeding tool for creating variation in

fruit crops. It provides an opportunity for the improvement of
traits like dwarf plant, earliness, tolerance and resistance to
various diseases and pests within short period of time. Mutant
identification or selection at the genotypic level, using new
technologies, has changed the way mutations are now used in
genetics and breeding in Fruit crops. In vitro culture combined
with induced mutation had been proven to speed up the
breeding program to produce genetic variations or for
multiplication. It also help in development of commercial
varieties in achieving the target of nutritional security.
THANK YOU