Int.J.Curr.Microbiol.App.

Sci (2017) 6(12): 3620-3633

International Journal of Current Microbiology and Applied Sciences

ISSN: 2319-7706 Volume 6 Number 12 (2017) pp. 3620-3633
Journal homepage: http://www.ijcmas.com

Review Article https://doi.org/10.20546/ijcmas.2017.612.418

Mutation Studies in Fruit Crops: A Review

Kunzang Lamo1*, Deep Ji Bhat1, Kiran Kour1 and Shivendu Pratap Singh Solanki2
1
Sher-e-Kashmir University of Agricultural Sciences and Technology, Jammu-180009, India
2
Department of Fruit Science, PAU, Ludhiana, Punjab, India
*Corresponding author

ABSTRACT

Mutation is sudden heritable changes in the DNA sequence that are not derived from
genetic segregation or recombination. Spontaneous mutation occurs at a very low
Keywords
frequency 1 in 10 lacs or 10-6. In fruit crops spontaneous bud mutation are more common
Mutation, called as bud sports. The occurrence of a large number of natural bud sports in citrus,
Spontaneous, mango, grapes etc. made the fruit breeders interested to breed through induced mutation.
Induced, Since, mutations bring about variation, they provide the ultimate basis for evolution of new
Improvement, forms, varieties or species. Mutations may result into deletion, inversion, translocation of
Fruits. chromosome and nucleotide base substitutions. Mutation can be induced artificially with
Article Info the help of various physical and chemical agents which are called mutagens. Most
commonly used Physical and chemical mutagens are gamma rays and EMS (Ethyl
Accepted: Methane Sulphonate) respectively. Dose inducing 25 to 50% lethality (LD 25-LD50) among
28 October 2017 mutated plants will result in the highest mutation rates. Perennial nature, long juvenile
Available Online: phase, heterozygosity, sexual incompatibilities etc in fruit crops limits their improvement
10 December 2017 through conventional breeding.

Introduction
Mutations are defined as sudden heritable has also been used in a wider sense to include
changes in the genetic material of an the exploitation of natural as well as
organism and in turn in its characters that are spontaneous mutants, and in the development
not derived from genetic segregation or of any variety possessing a known mutation
recombination (Van Harten, 1998). De Vries from whatever source. Mutants are the
used the word “sudden” to differentiate individuals carrying a mutation that may be
between subtle changes that could be revealed using molecular means or identified
explained by the normal processes of by phenotypic tools. Different types of mutant
recombination. The origins were in the can be generated using experimental
variants described in taxonomic treatises of mutagenesis. Mutagens these are the agent
wild and cultivated plants and in spontaneous use to induce mutation. Spontaneously arising
mutants (bud sports, etc) of value and interest mutations are very rare and random events in
in domestication and crop improvement. The terms of the time of their occurrence and the
term mutation breeding was coined to refer to gene in which they occur. In this way mutant
the deliberate induction and development of forms showing both large and small effects on
mutant lines for crop improvement. The term the phenotype arise for all kinds of traits.
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Many of the mutations may be deleterious large and small effects on the phenotype arise
making the organism less adapted to its for all kinds of traits. Many of the mutations
environment and some may even be lethal. may be deleterious making the organism less
Mutagenesis can be exploited experimentally adapted to its environment and some may
(experimental mutagenesis) by physical, even be lethal. Some may be neutral in their
chemical and biological means. The fruit effects and may confer no immediate
industry relies on a limited number of clonally advantage, but may help to generate a wide
propagated cultivars established on range of useful recombinant genotypes
recognized fruit quality parameters and through the subsequent generations. However,
consumer familiarity with the product, and is on rare occasions, some cultivars can
very reluctant to changes (Janick, 1992). This demonstrate unstable phenotypes resulting in
limits the use of cross-breeding in fruits, as a portion of the plant, sometimes extending to
fruit cultivars are generally highly whole branches, having different
heterozygous, and progenies from cross- characteristics. When these branches (bud
breeding express a large number of traits sports) are vegetatively propagated by clonal
which are different from those of the parents. techniques, the new phenotype is generally
Other specific problems (e.g. polyploidy, maintained leading to a new variety, often
incompatibility, apomixis, long juvenile exhibiting only one phenotypic character
period) can make obtaining useful different from the parent (Marcotrigiano
recombinants a laborious task (Broertjes, 1997). Horticulturist propagates bud sports
1977; Hansche and Beres, 1980). In contrast, because they continue to exhibit all of the
induced mutations change only one or a few other desirable characteristics of the parent
specific traits of an elite cultivar, and can (Franks et al., 2002). Bud sports are
contribute to fruit improvement without infrequent changes in phenotype affecting
upsetting neither the requirements of the fruit shoots of woody perennials but the molecular
industry nor the consumers. In fruit crops, basis of these mutations has rarely been
mutagenesis has already been used to identified (Table 1).
introduce many useful traits affecting plant
size, blooming time and fruit ripening, fruit When a mutation (plastidial, genic,
color, self-compatibility, self-thinning, and chromosomal, genomic) arises in a cell within
resistance to pathogens (Visser et al., 1971; a shoot apical meristem of a bud, the mutated
Janick and Moore, 1975; Donini, 1982; cell propagates mitotically and produces a
Broertjes and Van Harten, 1988; Spiegel-Roy, mutated sector (sectorial or mericlinal
1990; Spina et al., 1991; Brunner and Keppl, chimera) (D' amato 1977). During further
1991; Masuda et al., 1997; Janick and Moore, vegetative growth of the initial bud one or
1996; Van Harten, 1998; Sanada and Amano, more wholly mutated buds (solid mutant or
1998). sport may be produced. Mutated buds may
develop into an independent plant through
Types of mutation various naturally occurring modes of
vegetative propagation (tubers, tubercles,
Spontaneous mutation tuberous roots, bulbs, pseudobulbs, cormets,
stolons, rhizomes, root crowns, stem points,
Spontaneously arising mutations are very rare inflorescence bulbils, adventitious embryony
and random events in terms of the time of and apomictic seed).Obviously, sports can
their occurrence and the gene in which they also be propagated through true (amphimictic)
occur. In this way mutant forms showing both seed when the original plant reproduces

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sexually. Among the bud sports, the most shoot tip, leaf, ovules, protoplasts and
common and wide spread mutations are rhizomes. and more recently, the induction of
colour alterations in the red or purple mutations in vegetatively propagated plants is
anthocyanin content of fruit (Walker et al., becoming more efficient as scientists take
2006). Compared with conventional advantage of totipotency (ability of a single
hybridization breeding, bud sport is a cell to divide and produce all of the
consequence of genetic variation of somatic differentiated cells in an organism to
cells leading to the occurrence of qualitative regenerate into whole plants) using single
and quantitative phenotypic alteration in cells and other forms of in vitro cultured plant
plants, which can be observed in many tissues. Whereas chemical mutagens are
vegetatively propagating plants including preferably used to induce point mutations,
grapes (Liu et al., 2007). Bud mutants have physical mutagens induce gross lesions, such
been widely exploited by vine growers to as chromosomal abbreviation or
develop new cultivars of wine grapes and rearrangements. It is noteworthy that the
table grapes. Zhao et al., (2014) reported an frequency and types of mutations are direct
„Early-ripening Benitaka‟ grape, a bud sport results of the dosage and rate of exposure or
of the „Benitaka‟ (Vitis vinifera) cultivar that administration of the mutagen rather than its
exhibits a marked difference in sugar and type Mba et al., (2012). In the end, the choice
anthocyanin accumulation in ripening fruit of a mutagen will be based more often than
arises from a spontaneous mutation. Walker et not on the particular researcher‟s
al., (2006) reported that two new grape circumstances, such as safety of usage, ease
cultivars which were bud sports of Cabernet of use, availability of the mutagens,
Sauvignon namely Malian or Bronze effectiveness in inducing certain genetic
Cabernet berries and Shalistin or White alterations, suitable tissue, cost and available
Cabernet berries. Liu et al., (2007) reported a infrastructure among other factors. Tissue
spontaneous bud mutation in sweet orange culture has a potential for improving
(Citrus sinensis L. Osbeck) „Hong Anliu‟, effectiveness of mutation induction in several
which results in fruits with lycopene aspects. First of all, it offers a wide choice of
accumulation, low citric acid, and high plant material for treatment that mutation
sucrose was reported in a previous study. induction. In fact, structures that will give
origin to plants are composed of a few or even
Induced mutation of one cell. This means less risk of obtaining
chimaeric plants and a higher probability for
Agents of artificial mutations are called mutated cells to express the mutation in the
mutagens. They are generally grouped into phenotype (D‟ Amato, 1977). Tissue culture
two broad categories, namely chemical also allows for the handling of large
mutagens and physical mutagens (Mba et al., populations for mutagenic treatment,
2010). Traditionally, to induce mutations in selection, and cloning of selected variants. It
crops, planting materials are exposed to also offers the possibility to rapidly execute
physical and chemical mutagenic agents. the propagation cycles of subculture aimed to
Mutagenesis can be performed with all types separate mutated from non-mutated sectors
of planting materials, e.g. whole plants, (dissolving a chimera to obtain homo-histone)
usually seedlings, and in vitro cultured cells. (Ahloowalia, 1998). Despite these prospects
Nevertheless, the most commonly used plant and a number of research studies, only two
material is seed. Multiple forms of plant fruit cultivars derived from in vitro mutation
propagules, such as bulbs, tubers, corms, induction have been released to date i.e.

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cultivars of banana (Musa acuminata Colla) become ions therefore, the term ionizing
„Klue Hom Thong KU1‟ and „Novaria‟. radiation was coined. These ionizing
These poor results suggested that mutagenesis components of the electromagnetic include
in combination with tissue culture is either cosmic, gamma and X-rays. X-rays were the
ineffective or has yet to be exploited in fruit first to be used to induce mutations and
crops. The availability of tissue culture gamma radiation from radioactive cobalt
protocols for plant micropropagation and (60Co) is widely used. It has high penetrating
regeneration from cell and tissues for nearly potential and is hazardous. However, it can be
all of the fruits commercially most important used for irradiating whole plants and delicate
(George, 1993; Herman, 2000) calls for a materials, such as pollen grains.
wider use of in vitro mutation induction. The
fruit industry relies on a limited number of Use of gamma radiation
clonally propagated cultivars established on
recognized fruit quality parameters and The use of radiation technique with in vitro
consumer familiarity with the product, and is propagation over plants to out numbered
very reluctant to changes (Janick, 1992; vegetatively is an effective technique in terms
Mehlenbacher, 1995). This limits the use of of obtaining variation, quick proliferation of
cross-breeding in fruits, as fruit cultivars are mutants and obtaining disease-free mutants.
generally highly heterozygous, and progenies This combination has been successfully
from cross-breeding express a large number applied in date palm, apple and pineapple
of traits which are different from those of the (Anonymus, 2001). Sutarto et al., (2009) with
parents. Other specific problems (e.g. their 20 Gy and 40 Gy gamma application,
polyploidy, incompatibility, apomixis, long determined two mandarin and one pummelo
juvenile period) can make obtaining useful species to have potential in terms of being
recombinants a laborious task (Broertjes, seedless plants and offered field practices
1977; Hansche and Beres, 1980). In contrast, over these species. Yoshioka et al., 1999
induced mutations change only one or a few reported that gamma radiation induced
specific traits of an elite cultivar, and can mutants which are resistant to black spot
contribute to fruit improvement without disease, caused by Alternaria alternata in
upsetting neither the requirements of the fruit Japanese pear, is the most important and
industry nor the consumers. serious disease in Japanese pear. Nine
resistant mutants were selected from
Physical mutagens chronically irradiated „Nijisseiki‟. One of
these mutants was registered as „Gold
In the past 80 years, physical mutagens, Nijisseiki‟. A resistant mutant derived from
mostly ionizing radiations, have been used acutely irradiated dormant scions of „Shinsui‟
widely for inducing hereditary aberrations and was registered as „Kotobuki Shinsui‟. Four
more than 70% of mutant varieties were resistant mutants were selected from
developed using physical mutagenesis (Mba chronically irradiated „Osanijisseiki‟. One of
et al., 2012). Radiation is defined as energy them was registered as „Osa Gold‟. Moreover,
travelling through a distance in the form of mutations with a higher level of resistance
waves or particles. These are relatively high- than that of „Gold Nijisseiki‟ were induced
energy levels of electromagnetic (EM) from „Gold Nijisseiki‟ by acute and chronic
spectrum that are capable of dislodging gamma-ray irradiation. Khalil et al., (2011)
electrons from the nuclear orbits of the atoms developed sparse seeded mutant of kinnow
that they impact upon. The impacted atoms (Citrus reticulata Blanco) using gamma

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radiation. Dormant bud scions of Kinnow (1998) proposed the precise method for the
were subjected to gamma irradiation dose of determination of absorbed dose of radiation.
20 Gy which was best. The irradiated bud A concept of LD50 (lethal dose 50 %) is used
scions were grafted onto Rough lemon to refer the optimum dose to be used in the
rootstock, using the side-graft technique. experiment. By definition LD50 is the dose
Russo et al., (1981) carried out induced which causes 50 % lethality in the organism
mutations with gamma rays in seeded used for irradiation in defined time. It varies
„Monreal‟ Clementine mandarin (Citrus with the plant species, the type and status of
reticulate Blanco) and produced mutants with the material and the stage at which lethality is
fewer seeds per fruit. An early flowering measured. Generally, irradiated populations
putative mutant designated 'GN-60A' was are generated by using an LD50 dose treatment
identified among the regenerated plants and with a dose lower than LD50. To obtain a
obtained from gamma ray-treated populations mutant, the dose of the mutagen should be
in the glasshouses of the FAO/IAEA sufficiently high to increase the probability of
Agriculture and Biotechnology Laboratory, inducing a mutation.
Seibersdorf, Austria. The GN-60A clone has
been micropropagated and its vegetative However, dose should not be so high as to
progeny sent to Honduras, Australia, South cause damage to the cells or tissues resulting
Africa and Malaysia for field testing under in lethality. Since, induction of mutation is a
commercial plantation conditions. chance event, and recovery of a mutation is
dependent upon chance of the survival of that
In Malaysia, early flowering plants obtained individual plant, this strategy improves the
in the field were tissue cultured again to probability of obtaining a desirable mutant.
produce about 2000 plants for commercial Radiosensitivity varies with the species and
evaluation in the United Plantations Bhd. in the cultivar, with the physiological condition
September 1993. From this population a of plant and organs, and with the
selection, flowering about 10 weeks earlier manipulation of the irradiated material before
than the original parental clone (i.e. Grande and after mutagenic treatment (Briggs and
Naine), was launched under the name Novaria Konzak, 1977; D‟Amato, 1992) (Table 2). In
in 1995. Ram and Majumder (1987) plants which are sensitive to radiation, doses
developed a dwarf mutant of papaya known lower than LD50 are also used to reduce the
as Pusa Nanha through mutation breeding by mutation load (Shu et al., 2012).
treating the seeds of papaya strain Pusa 1-15
with 15 Kr gamma rays (Table 3). Chemical mutagens

Doses of mutagens Chemical mutagens were found to be highly

effective in inducing true gene mutations and
One of the first steps in mutagenic treatment the specificity of action could be investigated
is the estimation of the most appropriate dose through analysis of their reaction with
to apply. Radiation dose is expressed in rads different DNA base. There are different
(radiation-absorbed dose) which is equivalent chemical mutagens used to induce mutation in
to absorption of 100 ergs/g. The unit kilo rad fruit crops namely alkylating agents most
(kR which is 1,000 rads) which was in use useful for mutation Ethyl methanesulphonate
earlier is replaced by gray (Gy) which is (EMS), Diethyl sulphate (dES),
currently used. The two can be interconverted Ethyleneimine (EI), Ethyl nitroso urethane
as 1 kR is equivalent to 10 Gy. Neville et al., (ENU), Ethyl nitroso urea (ENH) and azides

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(Heslot, 1977). As compared with physical banana and grapes (Omar et al., 1989; Van
mutagens, chemicals may give rise to Harten, 1998; Predieri, 2001; Luan et al.,
relatively more gene mutations rather than to 2006 and Berenschot et al., 2008). Mutation
chromosomal changes. The assessment of induction in banana has been carried out by
LD50 for chemicals is determined by varying subjecting of shoot tips to mutagen followed
the concentration and duration of treatment, by regenerating of treated shoot tips (Predieri,
the solvent used (e.g. Dimethyl sulfoxide 2001). Bhagwat and Duncan (1997) also used
(DMSO), or the pH of the solution (Novak, chemical mutagens in banana (Musa spp.
1991). EMS was used for inducing mutations AAA group) to produce variants displaying
in banana by treating shoot tips and then resistance against fusarium wilt (Fusarium
regenerating adventitious buds. Among oxysporum f. sp. cubense). They determined
chemicals, ethyl methanesulphonate (EMS) 200 mM of EMS for 30 min as an optimal
belonging to the group of the alkylating dose and duration treatment. In vitro mutation
agents has been reported as a very effective induction for salt tolerance using EMS was
and efficient mutagen for creating of also reported in sweet potato (Luan et al.,
somaclonal variation in crop plants such as 2006).

Table.1 Identified spontaneous mutants in fruit crops

Crop Original variety Mutant cultivar Nature of Reference

mutation and
traits
Mango Rosado de Lea Rosica Bud sports Medina
Davis haven Haden Precocious, (1977)
regular bearer, Young and
and larger fruit Ledin (1954)
size
Large fruit size
Banana Highgate Gros michel Sports, semi Daniells
Motta Poovan Poovan dwarf (1990)
Sports
Grapefruit Foster Hudson Bud sports, deep Soost and
red flesh Cameron
(1975)
Pear Clapp‟s Favourite Starkrimson Bud sports, Bishop
spotting of (1966)
coloured
Mandarin Owari Pongan Clausellina Pongan Bud sport
86-1 -
Navel Orange Bahia Baianinha Limb sports
Washington Navelina, Limb sports -
Navelate,Marrs,
Leng, Autumn Gold,
Powell Summer,
Winter Red
(Ray, 2002)

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Table.2 Some examples of radiosensitivity of tissue in fruit crops

Species Mutagen Plant material LD50 (Gy) Reference

Apple gamma rays leaves >10–<20 Predieri and Fasolo
(1989)
Japanese plum gamma rays Shoots 30 Predieri and Gatti (2000)
Strawberry gamma-rays Shoot clumps >50–<100 Jain (1997)
Kiwi gamma-rays leaves 40–60 Sammarcelli-Ollitraut and
Legrave (1991)
Banana gamma-rays Shoot tip 30–35 (Triploid)

Table.3 Achievements through mutation breeding

Species Cultivar released Year Mutagen Trait improved

Apple Golden Haidegg 1986 Gamma rays Large fruit size
Lysgolden 1970 Gamma rays Rust resistance
Courtavel 1972 Gamma rays Shortness
Senbatsu-Fuji-2-Kei 1985 gamma rays Fruit colour
Peach Mangnif 135 1986 Gamma rays Fruit size
Plovdiv 6 1981 Gamma rays yield
Banana Novaria 1993 Gamma rays Earliness
Klue Hom Thong KU1 1985 Gamma rays, in vitro Bunch size
Orange/ Valencia 2 INTA 1987 X-rays Fruit set, quality
mandarin
Eureka 22 INTA 1987 X-rays Fruit set, quality
Papaya Pusa Nanha 1986 Gamma rays Dwarfness
Pear Kotobuki shinsui 1996 Gamma rays Disease resistance
Apricot Early Blenheim 1979 ThN Earliness
Grape Fikreti 1986 Gamma rays Earliness
Almond Supernova 1987 Gamma rays Fruit size
Plum Spurdente-Ferco 1988 Gamma rays Earliness
Sour Plodorodnaya Michurina 1977 X-rays Fruit set
cherry Plodorodnaya Orlovskoi 1979 Gamma rays Dwarfness
Karlik Samorodka 1979 Gamma rays Dwarfness
sweet Burlat C1 1983 Gamma rays Compact growth
cherry Ferrovia spur 1992 X-rays Dwarfness
Compact Stella 35B11 1974 X-rays Compact growth
(Predieri, 2001).

Table.4 DNA markers for genetic diversity assessment in fruit crops

Fruit Marker Type References

Apple AFLP and RAPDs Coart et al., (2003); Botez et al., (2009); Sestras et al., (2009)
Avocado Mini satellite DNA Ashworth et al., (2003)
Banana RAPDs Brown et al., (2009)
Citrus RFLP Durham et al., (1992)
Pistachio Mini satellite marker Riaz Ahmad et al., (2003)
Pear SSRs and AFLP Sisko et al., (2009)
Cashew RAPD and ISSR Thimmappaiah et al., (2009)

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Colchicine treatment 1991) have also been reported. Similarly in

apple rootstocks M26 and MM106 (Malus
Colchicine has been used on small fruits with pumila Mill.). Peach somaclones resistant to
the aim of improving yield parameters (e.g. the root-knot nematode (Meloidogyne
fruit size) or for using polyploids in cross- incognita Kofoid and White) have been
breeding. Colchicine has been widely used to obtained as well (Hashmi et al., 1995). Khalil
increase ploidy level (Van Harten, 1998). et al., (2011) produce Bingtang sweet orange
This is of vital importance for banana cross- somaclones tolerant to citrus canker disease
breeding (Van Duren et al., 1996). Colchicine by in vitro mutagenesis with 0.5 % of EMS.
was used for inducing ploidy doubling in Somaclones resistant to canker disease was
octoploid tissue cultured strawberry later known as DG-2. Kuksova et al., (1997)
(Fragaria × ananassa Duch.) (Predieri et al., reported that grapevine somaclones
1989). Shoots were treated in a liquid solution possessing field resistance to Botrytis cinerea
with concentrations ranging from 0.65 to 12.5 and Plasmopara viticola have also been
mM for periods varying from five to nine found. With respect to the issue of resistance
days. Colchicine was used to obtain to abiotic stress, genotypes with increased
tetraploids of banana by immersing shoot tips tolerance to salinity were obtained from
of the diploid clone „SH- 3362‟ in a liquid MS somaclonal variants in Citrus species (Ben-
medium (Murashige and Skoog, 1962) Hayym and Kochba, 1983; Ben- Hayym and
containing 0–1% (2.5 mM) colchicine. Stable Goffer, 1989).
autotetraploids were established in the field,
although some chimerism and reversion to Application of molecular for the
diploid were recorded (Hamill et al., 1992). detectation of mutation in fruit crops
Wu hu et al., (2012) reported that colchicine-
induced red-fleshed Actinidia chinensis Before 21st century, mutants were mostly
autotetraploids produced from three female selected by observing phenotypes of
selections („Hort 22 D‟, Selection 1 and individual plants in mutated populations
Selection 2) and one male (Selection 3). Sun treated with chemical or physical mutagens.
et al., 2009 reported that a European pear Concurrently, various molecular and
(Pyrus communis L.) cultivar „Fertility‟ was genomics tools have been developed for the
evolved by in vitro colchicine treatment of detection of genetic variants including single
leaf explants. nucleotide polymorphism (SNP). The
integration of genomics information and
Somaclonal variation molecular tools has resulted in the
development of various molecular approaches
Somaclonal variation has led to the selection that can be use in the screening for mutants in
of several variants with increased resistance large populations derived from radiation and
to pests, diseases, and herbicides (Brar and chemical mutagenesis.
Jain, 1998), and can be considered as a tool
for inducing variation in fruit improvement Techniques
(Bouharmont, 1994; Karp, 1995;
Hammerschlag et al., 1995). In strawberry, DNA marker techniques can also be used
somaclones resistant to Fusarium oxysporum widely in plant mutation breeding and genetic
f. sp. fragariae (Toyoda et al., 1991), to research for increasing both the efficiency and
Alternaria alternate (Takahashi, 1993), and to efficacy of mutation techniques. They can be
Phytophthora cactorum (Battistini and Rosati, used for tracing the pedigree of induced

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mutants and tagging important mutations. markers such as proteins and enzymes can be
Consequently, closely linked markers of used as efficient markers in crop
mutant traits can be used for MAS, improvement, AFLP typically allows the
pyramiding and cloning of mutant genes. analysis of dozens of DNA markers
However, the way they are used is somewhat simultaneously (Table 4). Combinations of
different from conventional plant breeding primers with alternate supplementary bases
programmes. Some biochemical markers such allow many different sub-sets of DNA marker
as proteins and enzymes can be used as combinations, or genomic representations, to
efficient markers in crop improvement, AFLP be analyzed, a feature that makes this
is a technique that combines aspects of both technique very versatile. Like RAPD markers,
RFLP and DNA amplification using the AFLP markers are either present or absent.
polymerase chain reaction (PCR). AFLP AFLP band intensities can be quantified and
typically allows the analysis of dozens of scored as co-dominant marker, especially
DNA markers simultaneously. Combinations between samples of closely related
of primers with alternate supplementary bases individuals such as backcross populations to
allow many different sub-sets of DNA marker discriminate heterozygous loci.
combinations, or genomic representations, to
be analyzed, a feature that makes this Mutation is important breeding tool for
technique very versatile. Like RAPD markers, creating variation in fruit crops. It provides an
AFLP markers are either present or absent. oppurtunity for the improvement of traits like
AFLP is normally used as a genetic dwarf plant, earliness, tolerance and
fingerprinting method or when large numbers resistance to various diseases and pests within
of markers are required from which candidate short period of time. Mutant identification or
markers linked to a target can be selected. selection at the genotypic level, using new
technologies, has changed the way mutations
For sheer density of DNA markers, single are now used in genetics and breeding in Fruit
nucleotide polymorphisms (SNPs) offer one crops. In vitro culture combined with induced
of the richest sources. SNPs are increasingly mutation had been proven to speed up the
used as DNA markers because techniques are breeding program to produce genetic
becoming more widespread, cost effective variations or for multiplication. It also help in
and automated and the genome sequence development of commercial varieties in
information of major crops are becoming achieving the target of nutritional security.
increasingly available for use. Since many
naturally occurring mutations are single References
nucleotide polymorphisms (SNPs), various
technologies have been developed for SNP Acquaah, G. Principles of plant genetics and
detection, most of which are also useful for breeding. Chichester: Wiley-Blackwell;
the detection of small insertion/deletions 2006.
(indels). They can be used for tracing the Amanda, R., Walker, E. L. and Robinson, S. P.
pedigree of induced mutants and tagging 2006. Two new grape cultivars, bud
important mutations. Consequently, closely sports of Cabernet Sauvignon bearing
linked markers of mutant traits can be used pale-coloured berries, are the result of
for MAS, pyramiding and cloning of mutant deletion of two regulatory genes of the
genes. However, the way they are used is berry colour locus. Plant Molecular
somewhat different from conventional plant Biology, 62:623- 635.
Anonymus, “In Vitro Techniques for Selection
breeding programmes. Some biochemical
of Radiation Induced Mutations Adapted

3628
 Int.J.Curr.Microbiol.App.Sci (2017) 6(12): 3620-3633

to Adverse Environmental Conditions”. Mutations in Crop Improvement (pp.

Proceedings of a final Research Co- 15–37). Kluwer Academic Publishers,
ordination Meeting organized by the Dordrecht.
Joint FAO/IAEA Division of Nuclear Briggs, R. W. and Constantin, M. J. 1977.
Techniques in Food and Agriculture and Radiation types and radiation sources.
held in Shanghai, China, 17–21 August In: Manual on Mutation Breeding.
1998, 2001. Second edition. Technical Reports Series
Ashworth, V. E. and M. T. Clegg. 2003. No. 119: 7-20). IAEA, Vienna.
Microsatellite markers in avocado Broertjes, C. 1977. General considerations,
(Persea mericana Mill.): Genealogical breeding methods and selection of
relationships among cultivated avocado parents. In: Manual on Mutation
genotypes Journal of Heredity, 94:407- Breeding. Second edition. Technical
415. Reports Series, 119: 159–160. IAEA,
Battistini, C. and Rosati, P. 1991. In vitro Vienna.
evaluation of somaclonal strawberry Broertjes, C. and Van Harten, A. M. 1985.
(Fragaria × ananassa „Brighton‟) Single cell origin of adventitious buds.
variants for susceptibility to Euphytica, 34: 93–95.
Phytophthora cactorum In: Dale, A. and Brown, N., S. Venkatasamy, G. Khittoo, T.
Lubby, D. (eds.). The Strawberry Into Bahorun and S. Jawaheer. 2009.
the 21st Century (pp 121–123). Timber Evaluation of genetic diversity between
Press, Portland. 27 banana cultivars (Musa spp.) in
Ben-Hayyim, G. and Goffer, Y. 1989. Plantlet Mauritius using RAPD markers. African
regeneration from NaCl selected salt- Journal of Biotech, 8:1834-1840.
tolerant callus culture of Shamouti Brunner, H. and Keppl, H. 1991. Radiation
oranges (Citrus sinesis L. Osbeck). Plant induced apple mutants of improved
Cell Reports, 7: 680–683. commercial value. In: Plant Mutation
Ben-Hayyim, G. and Khocba, J. 1983. Aspects Breeding for crop Improvement, 1: 547–
of salt tolerance in a NaCl sodium 552. IAEA, Vienna.
chloride-selected stable cell line of Cleggett, M. D. 2002. Malian. Plant Variety
Citrus sinesis Shamouti oranges. Plant Journal, 15:85–86.
Physiology, 72: 685–690. Cleggett, M. D. 2003. Variety: „Shalistin‟. Plant
Boss, P. K., Davies C, Robinson, S. P. 1996. Variety Journal, 16:64–65.
Anthocyanin composition and Coart, E., Vekemans, X., Smulders, M. J. M. I.,
anthocyanin pathway gene expression in Wagner, J., Huylenbroeck, V.,
grapevine sports differing in berry skin Bockstaele, V. and Roldan-Ruiz. I. 2003.
colour. Australian Journal of Grape Genetic variation in the endangered wild
Wine Research, 2:163–170. apple (Malus sylvestris L. Mill.) in
Bouharmont, J. 1994 Application of somaclonal Belgium as revealed by amplified
variation and in vitro selection to plant fragment length polymorphism and
improvement. Acta Horticulturae, 355: microsatellite markers. Molecular
213–218. Ecology, 12:845-857.
Bowers, J. E. and Meredith, C. P. 1997. The D‟amato, F. 1965. Cytological and genetic
parentage of a classic wine grape, aspects of aging. In: Proceedings XII
Cabernet. International Congress Genetics, pp.
Brar, D. S. and Jain, S. M. 1998. Somaclonal 285-295, Pergamon Press., Oxford.
variation: mechanism and applications in D‟amato, F. 1992. Induced mutations in crop
crop improvement. In: Jain, S. M., Brar, improvement: Basic and applied aspects.
D. S. and Ahloowalia, B. S. (eds) Annals of Agricultural and
Somaclonal Variation and Induced Environmental Medicine, 122: 31–60.
3629
 Int.J.Curr.Microbiol.App.Sci (2017) 6(12): 3620-3633

Daniells, J., Davis, D., Peterson, R. and Pegg, Hamill, S. D., Smith, M. K. and Dodd, W. A.
K. 1995. Goldfinger: not as resistant to 1992. In vitro induction of banana
sigatoka or yellow sigatoka as first autotetraploids by colchicine treatment
thought. Infomusa, 4(1): 6. of micropropagated diploids. Australian
Deane, C. R., Fuller, M. F. and Dix, P. J. 1995. Journal of Botany, 40: 887–896.
Selection of hydroxyproline resistant Hammerschlag, F. A., Ritchie, D., Werner, D.,
proline accumulating mutants of Hashmi, G., Krusberg, L., Meyer, R. and
cauliflower. Euphytica, 85: 329-334. Huettel, R. 1995. In vitro selection of
Deng, Z. N., Xu, L., Li, D. Z., Long, G. Y., Liu, disease resistance in fruit trees. Acta
L. P., Fang, F. and Shu, G. P. 2010. Horticulturae, 392: 19–26.
Screening citrus genotypes for resistance Hansche, P. E. and Beres, W. 1980. Genetic
to canker disease (Xanthomonas remodeling of fruit and nut trees to
axonopodis pv. citri). Plant Breeding, facilitate cultivar improvement.
129(3): 341–345. HortScience, 15: 710–71.
Derman, H. 1947. Histogenesis of some bud Hashmi, G., Hammerschlag, F. A., Huettel, R.
sports and variegations. American N. and Krusberg, L. R. 1995. Growth,
Society of Horticultural Science, 50: 51– development and response of peach
73. somaclones to the root-knot nematode,
Donini, B. 1982. Mutagenesis applied to Meloidogyne incognita. Journal of
improve fruit trees: techniques, methods American Society for Horticultural
and evaluation or radiation induced Science, 120: 932–937.
mutations. In: Induced Mutations in Hearn, C. J. 1984. Development of seedless
Vegetatively Propagated Plants, II (pp orange and grapefruit cultivars through
29-36). IAEA, Vienna. seed irradiation. American Society of
Durham, R. E., Liou, P. C., Gmitter, F. G. and Horticultural Sciences, 109: 270- 273.
Moore, G. A. 1992. Linkage of Hearn, C. J. 1986. Development of seedless
restriction fragment length grapefruit cultivars through bud wood
polymorphism and isozymes in Citrus. irradiation. American Society of
Theoretical and Applied Genetics, Horticultural Sciences, 11(1): 304-306.
84:39-48. Hensz, R. A. 1971. Star Ruby, a new deep-red-
Franks, T., Botta, R. and Thomas, M. R. 2002. fleshed grapefruit variety with distinct
Chimerism in grapevines: implications tree characteristics. Journal Rio Grande
for cultivar identity, ancestry and genetic Valley Horticultural Society, 25: 54-58.
improvement. Theoretical and Applied Herman, E. B. (ed.). 2000. Regeneration and
Genetics, 104:192–199. Micropropagation: Techniques, Systems
Freisleben, R.A. and Lein, A. 1944. and Media 1997–1999. Agritech
Möglichkeiten und praktische Consultants, Shrub Oak.
Durchführung der Mutationszüchtung. Heslot, H. 1977. Review of main mutagenic
Kühn-Arhiv. 60: 211-22. compounds. In: Manual on Mutation
Ge, H., Li, Y., Fu, H., Long, G., Lu., L. and Li, Breeding. Second edition. IAEA.
R. D. Z. 2015. Production of sweet Vienna. Technical Reports Series No.,
orange somaclones tolerant to citrus 119: 51–58.
canker disease by in vitro mutagenesis IAEA, 1997. Report 2nd FAO/IAEA Research
with EMS. Plant Cell Tissue Organ Co-ordination Meeting on in vitro
Culture, 123:29–38. techniques for selection of radiation
George, E. F. 1993. Plant Propagation by Tissue induced mutants adapted to adverse
Culture (pt. 1. Technology; pt. 2. In environmental conditions. International
practice). Exegetics Ltd., Edington, Atomic Energy Agency, Vienna.
Wilts.
3630
 Int.J.Curr.Microbiol.App.Sci (2017) 6(12): 3620-3633

Jain, S. M. 1997. Creation of variability by line in sugar and acid metabolism at the
mutation and tissue culture for fruit ripening stage. Science China Life
improving plants. Acta Horticulturae, Sciences, 50: 511–517.
447: 69–77. Luan, Y. S., Zhang, J., Gao, X. R. and An, L. J.
Janick, J. 1992. Introduction. In: 2007. Mutation induced by ethyl
Hammerschlag, F. A. and Litz, R. E. methane sulphonate (EMS), in vitro
(eds.) Biotechnology of Perennial Fruit screening for salt tolerance and plant
Crops. (pp. xix–xxi). C.A.B. regeneration of sweet potato (Ipomoea
International, Wallingford, U.K. batatas L.). Plant Cell Tissue Organ
Janick, J. and Moore, J. N. 1975. Advances in Culture, 88:77–81.
fruit breeding. Purdue University Press, Mak, C. 1995. “NOVARIA - A new banana
West Lafayette. mutant induced by gamma irradiation”,
Kamile, U. and Nasircilar, A. G. 2015. Report of the first FAO/IAEA Research
Developments of gamma ray application coordination meeting on “Cellular
mutation breeding studies in recent biology and biotechnology including
years. International Conference on mutation techniques for creation of new
Advances. useful banana genotypes”, Vienna. Pp.
Karsinah, N. L., Indriyani, P. and Sukartini. 81-94.
2012. Effect of gamma irradiation on the Marcotrigiano, M. 1986. Origin of adventitious
growth of mango grafted material. shoots regenerated from cultured tobacco
Agricultural Biological and leaf tissue. American Journal of Botany,
Environmental Sciences, Pp. 22-23. 73: 1541–1547.
Kidner, C., Sundaresan, V., Roberts, K. and Marcotrigiano, M. 1997. Chimeras and
Dolan, L. 2000. Clonal analysis of the variegation: patterns of deceit.
Arabidopsis root confirms that position, Hortscience, 32:773–784.
not lineage, determines cell fate. Planta, Masuda, T., Yoshioka, T., Inoue, K., Murata,
211:191–199. K., Kitagawa, K., Tabira, H., Yoshida,
Kuksova, V. B., Piven, N. M. and Gleba, Y. Y. A., Kotobuki, K. and Sanada, T. 1997.
1997. Somaclonal variation and in vitro Selection of mutants resistant to black
induced mutagenesis in grapevine. Plant spot disease by chronic irradiation of
Cell Tissue and Organ Culture, 49: 17– gamma rays in Japanese pear
27. „Osanijisseiki‟. Journal of Japanese
Liu, L. P., Qu, J. W., Li, N., Li, D. Z., Long, G. Society for Horticultural Science, 66:
Y. and Deng, Z. N. 2012. In silico 85–92.
characterization and molecular Mba, C., Afza, R. and Bado, S. 2010. Induced
modelling of in Xanthomonas mutagenesis in plants using physical and
axonopodis Implications for primary chemical agents. In: Davey, M. R.,
metabolism or virulence. Plant Omics Anthony, P., editors. Plant cell culture:
Journal, 5(5): 494–502. essential methods. Chichester: John
Liu, Q., Xu, J., Liu, Y. Z., Zhao, X. L., Deng, Wiley and Sons, Ltd. pp. 111-130.
X. X., Guo, L. L. and Gu, J. Q. 2007. A Mba, C., Afza, R. and Shu, Q. Y. 2012.
novel bud mutation that confers Mutagenic radiations: X-rays, ionizing
abnormal patterns of lycopene particles and ultraviolet. In: Shu, Q. Y.,
accumulation in sweet orange fruit Forster, B. P., Nakagawa, H., editors.
(Citrus sinensis L. Osbeck). Journal of Plant mutation breeding and
Experimental Botany, 58: 4161–4171. biotechnology. Wallingford: CABI; p.
Liu, Y. Z., Xiong, J. J. and Deng, X. X. 2007. 83-90.
Difference of a citrus late-ripening
mutant (Citrus sinensis) from its parental
3631
 Int.J.Curr.Microbiol.App.Sci (2017) 6(12): 3620-3633

Medina, J. P. 1977. Rosica a new mango variety improvement. Proceedings International

selected in Ica. Peru. Fruit var. Journal, Society of Citriculture, 1: 68-70, Tokyo,
31: 88-9. Japan.
Mehlenbacher, S. A. 1995. Classical and Sammarcelli-Olliraut, F. and Legave, J. M.
molecular approaches to breeding fruit 1991. In vitro irradiation of the cultivar
and nut Crops for disease resistance. Hayward. Acta Horiculturae, 297: 115–
HortScience, 30: 466–477. 122.
Murashige, T. and Tuker. D. P. H. 1969. Sanada, T. and Amano, E. 1998. Induced
Growth factor requirement of citrus mutation in fruit trees. In: Somaclonal
tissue culture. Proceeding of the 1st Variation and Induced Mutations in Crop
International Citrus Symposium, Improvement. In: Jain, S. M., Brar, D. S.
3:1155–1169. and Ahloowalia, B. S. (eds.) Somaclonal
Novak, F. J. 1991. In vitro mutation system for Variation and Induced Mutations in Crop
crop improvement. In: Plant Mutation Improvement (pp. 401–409).
Breeding for Crop Improvement, 2: 327– Sharma, D. K. and Majumder, P. K. 1985.
342. IAEA, Vienna. Studies on the inheritance of mango. 2nd
Omar, M. S., Novak, F. J. and Brunner, H. International Symposium. Mango,
1989. In vitro action of ethyl methane Bangalore. 10-20.
sulphonate on banana shoot tips. Scientia Shu, Q. Y., Forster, B. P. and Nakagawa, H.
Horticulturae, 40: 283–295. 2011. Plant Breeding and Genetics
Penna, S., Mirajkar, S. and Bhagwat, S. G. Section Joint FAO/IAEA Division of
2015. Induced mutation and crop Nuclear Techniques in Food and
improvement. Plant Biology and Agriculture International Atomic Energy
Biotechnology, 1. Agency.
Penna, S., Vitthal, S. B. and Yadav, V. P. 2012. Sisko, M., Javornik, B., Siftar, A. and Ivancic.
In vitro Mutagenesis and selection in A. 2009. Genetic relationships among
Plant Tissue Culture and their prospects Slovenian pears assessed by molecular
for crop improvement. Global Science markers. Journal of American Soiety for
Books. Horticultural Scences, 134:97-108.
Predieri, S. and Gatti, E. 2000. Effects of Soost, R. K. and Cameron, J. W. 1975. Citrus
gamma radiation on plum (Prunus (In) Advances in Fruit Breeding (Janic,
salicina Lindl.) „Shiro‟. Advances in J. and Moore, J. N. Eds.), Purdue,
Horicultural Science, 14: 215–223. University Press, West Lafayett, Indiana,
Predieri, S. and Govoni, M. 1998. In vitro pp. 507-540.
propagation of compact pear clones. Spiegel, R. P. 1990. Economic and agricultural
Acta Horticulturae, 475: 127–132. impact of mutation breeding in fruit
Predrieri, S. 2001. Mutation induction and trees. Mutation Breeding Review, 5: 1–
tissue culture in improving fruits. Plant 26.
Cell Tissue Organ Culture, 64: 185-210. Spiegel, R. P. and Aliza, V. 1989. Induced
Ray, P. k. 2002. In: Breeding Tropical and mutation in Citrus Proceeding of the 6th
Subtropical Fruits. Narosa Publishing International Congress of SABRAO.
House, New Delhi. 770-773.
Riaz, A., Ferguson, L. and Stephen, M. S. 2003. Spina, P., Mannino, P., Reforgiato, R. G. and
Identification of Pistachio (Pistacia vera Starrantino, A. 1991. Use of mutagenesis
L) Nuts with microsatellite markers. at Istituto Sperimentale per
Journal of American Society for Agrumicoltura, Acireale. In: Plant
Horticultural Science, 128: 898-903. Mutation Breeding for Crop
Russo, F., Donini, B. and Starrantino, S. 1981. Improvement, 1. pp. 257–261. IAEA,
Mutagenesis applied for citrus Vienna
3632
 Int.J.Curr.Microbiol.App.Sci (2017) 6(12): 3620-3633

Sun, Q., Sun, H., Li, L. and Bell, R. 2009. In derived from callus of strawberry. Plant
vitro colchicine-induced polyploid Cell Reports, 10: 167–170.
plantlet production and regeneration Van Harten, A. M. 1998. Mutation breeding:
from leaf explants of the diploid pear theory and practical applications.
(Pyrus communis L.) cultivar, „Fertility‟. Cambridge University Press, Cambridge,
Shandong Institute of Pomology, Taian, pp. 137–158.
Shandong 271000, P.R. China 2 Visser, T., Verhaegh, J. J. and De Vries, D.
Appalachian Fruit Research Station, 1971. Pre-selection of compact mutants
USDA, ARS, Kearneysville, WV 25430- induced by X-ray treatment in apple and
2771. pear. Euphytica, 20: 195–207.
Sutarto, I., Agisimanto, D. and Supriyanto, A. Walker, A. R., Lee, E. and Robinson, S. P.
2009. “Development of Promising 2006. Two new grape cultivars, bud
Seedless Citrus Mutants through Gamma sports of Cabernet Sauvignon bearing
Irradiation”, Q.Y. Shu (ed.), Induced pale-coloured berries, are the result of
Plant Mutations in the Genomics Era. deletion of two regulatory genes of the
Food and Agriculture Organization of berry colour locus. Plant Molecular
the United Nations, Rome, pp. 306-308. Biology, 62: 623–635.
Takahashi, H. 1993. Breeding of strawberry Wu, J-H., Ferguson, A. R. and Murray, B. G.
cultivars resistant to Alternaria black 2011. Manipulation of ploidy for
spot of strawberry (Alternaria alternata kiwifruit breeding: in vitro chromosome
strawberry pathotype) Bull. Akita Pref. doubling in diploid Actinidia
Coll. Agric. 19: 1–44 (Curr. Cont. chinensis Planch. Plant Cell. Tissue and
Abstract). Organ Culture, 6: 503–511.
Thimmappaiah, W. G., Santhosh, S. D. and Ye, W., Qin, Y., Ye Z, T., Silva, J. A., Zhang,
Melwyn, G. S. 2009. Assessment of L., Wu, X., Lin, S. and Hu, G. 2009.
genetic diversity in cashew germplasm “Seedless Mechanism of a New
using RAPD and ISSR markers. Scientia Mandarin Cultivar „Wuzishatangju‟
Horticulturae, 120: 411-417. (Citrus Reticulata Blanco)”. Plant
Thomas, M. R. and Scott, N. S. 1991. Sciences, 177(1): 19-27.
Microsatellite repeats in grapevine reveal Yoshioka, T., Masuda, T., Kotobuk, K., Sanada,
DNA Polymorphisms when analyzed as T. and Ito, Y. 1999. Gamma-Ray-
Sequence- Tagged Sites (STSs). Theory Induced Mutation Breeding in Fruit
of Applied Genetics, 86: 985–990. Trees: Breeding of Mutant Cultivars
Thompson, M. M. and Olmo, H. P. 1963. Resistant to Black Spot Disease in
Cytohistological studies of cytochimeric Japanese Pear, Institute of Radiation
and tetraploid grapes. American Journal Breeding, National Institute of
of Botany, 50: 901–906. Agrobiological Resources (NIAR)
Toyoda, H., Horikoshi, K., Yamano, Y. and (Ohmiya, Naka, Ibaraki, 319-2293
Ouchi, S. 1991. Selection of Fusarium Japan).
wilt disease resistance from regenerant

How to cite this article:

Kunzang Lamo, Deep Ji Bhat, Kiran Kour and Shivendu Pratap Singh Solanki 2017. Mutation
Studies in Fruit Crops: A Review. Int.J.Curr.Microbiol.App.Sci. 6(12): 3620-3633.
doi: https://doi.org/10.20546/ijcmas.2017.612.418

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