Int.J.Curr.Microbiol.App.Sci (2017) 6(11): 1902-1912

International Journal of Current Microbiology and Applied Sciences ISSN: 2319-7706 Volume 6 Number 11 (2017) pp. 1902-1912 Journal homepage: http://www.ijcmas.com

Review Article https://doi.org/10.20546/ijcmas.2017.611.226

Biotechnology a Modern Tool for Fruits Production - A Review

Anuradha1*, Subhash Chander2 and Arvind Malik1

1Department of Horticulture, CCS Haryana Agricultural University, Hisar-125004, Haryana, India 2Division of Fruit Crops, ICAR-IIHR, Bangalore-560089, Karnataka, India *Corresponding author

ABSTRACT

The problem of long juvenility, incompatibility and sterility are the biggest hindrances for the improvement of fruit crops through conventional fruit breeding. To mitigate the biotic and abiotic stresses, improvement is desired in already cultivated cultivar for one or few traits. In that regard, the biotechnological tools will prove quite useful to expedite the rate

of fruit crop improvement . For production of quality planting material by tissue culture is K e yw or ds commercialized in banana and strawberry. Biotechnological tools such as somaclonal variation and in vitro mutagenesis are the choicest method for varietal development in Tissue culture, in vitro, Somaclonal banana. GCTCV-119 and Novaria are the result of somaclonal variation and in vitro variation, Embryo mutagenesis, respectively. Somatic hybridization has been explored for the rootstock and rescue, Molecular scion improvement of citrus. Somatic hybridization research of almost two decades in marker. citrus is yielding fruit now. Embryo rescue has helped in accelerating the pace of

Article Info stenospermocarpic improvement and ‘Sweet Scarlet’ and ‘’ are two of the seedless table grapevines, which were released using this technique. Transgenic variety Accepted: Rainbow and Sun Up carrying resistance to PRSV are being commercially cultivated in 15 September 2017 Hawaii of USA. With the introduction of concept of cisgenics, the genetically transformed Available Online: fruits are likely to get public acceptance. Molecular marker aid great help in early 10 November 2017 diagnosis of sex of dioecious fruit plant like date palm and papaya. Presently marker

linked to resistance for Dagger nematode and pierce disease at university of California in the breeding programs are routinely used for grapevine rootstock improvement. Genome sequencing has unveiled the mystery about various genes, the phylogenetic relationship

and evolution history of sequenced fruit crop.

Introduction

The requirement of fruits is increasing tools have revolutionized the entire crop proportionally with the increasing population. improvement programmes by providing new Although conventional plant breeding strains of plants, supply of planting material, techniques have made considerable progress more efficient and selective pesticides and in the development of improved varieties, improved fertilizers. Fruit crops are they have not been able to keep pace with the conventionally propagated through cutting, increasing demand for fruits. Therefore an budding, grafting, layering etc. but these immediate need is felt to integrate techniques are time consuming and biotechnology to speed up the crop cumbersome. Plant tissue culture provides an improvement programmes. Biotechnological effective measure of such problems of fruit 1902

Int.J.Curr.Microbiol.App.Sci (2017) 6(11): 1902-1912 crop multiplication. There is need of efficient Micropropagation technology which facilitates faster multiplication of propagules with potent Micropropagation of almost all the fruit crops regeneration. In recent years, synthetic seed is possible now. Production of virus free technology using encapsulation of non- planting material using meristem culture has embryogenic propagules derived under in been made possible in many horticultural crop vitro condition has become an important asset Strawberry is perhaps first fruit crop in which to micropropagation. Many genetically micropropagation technique has been modified fruits and vegetables are already in standardized (Sharma and Singh, 1999). In the market in developed countries. It is a new vitro plants are more uniform, produce higher aspect of biological and agricultural science number of runners, have better survival in the which provides new tools and strategies in the field, and the fruit yield increase in 24% than struggle against world’s food production plants propagated by the traditional method problem. (Kikas et al., 2006). Banana is commercially propagated through tissue culture. The future Biotechnological tool in fruit crop of banana industry in the country is totally improvement dependent upon control of BBTV disease. The ultimate choice is productions of disease- Tissue culture free banana plants through in vitro techniques to replace infected fields. There are number of It is one of the most widely used techniques fruit crop in which propagation method is for rapid asexual in vitro propagation. This sexual. Some of them are dioecious i.e. male technique is economical in time and space and female flower borne on different flowers. affords greater output and provides disease So through commercial point of view it is free and elite propagules. desirable to produce more female plant which, are the ones which bear fruits, this is possible It also facilitates safer and quarantined only through tissue culture techniques. The movements of germplasm across nations. grafting a very small shoot tip excised from When the traditional methods are unable to an elite mother tree grafted onto a decapited meet the demand for propagation material this rootstock seedling grown under aseptic technique can produce millions of uniformly condition. Murashige et al., (1972) made the flowering and yielding plants. first attempt in citrus and subsequently it was Micropropagation technology ensures true to refined by Navarro et al., (1975). type, rapid and mass multiplication of plants that possesses special significance in Embryo rescue vegetatively propagated plant species. This technology has witnessed a huge expansion Embryo rescue is another area, where plant globally. breeders are able to rescue their crosses, which would otherwise abort. Culture of Some basic techniques of tissue culture, such excised embryos of suitable stages of as anther/microspore culture, somaclonal development can circumvent problems variation, embryo culture, and somatic encountered in post zygotic incompatibility. hybridization, are being exploited to generate This technique is highly significant in useful genetic variability for obtaining intractable and long duration horticultural incremental improvement in commercial species. Ovule culture in (Singh et al., cultivars. 1991) aids in developing hybrids, even in

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Int.J.Curr.Microbiol.App.Sci (2017) 6(11): 1902-1912 seedless grapes. Embryo rescue has helped in in rootstock improvement, production of accelerating the pace of stenospermocarpic tetraploid rootstock for greater dwarfing grape improvement and ‘Sweet Scarlet’ and effect, which is used in HDP. Gene ‘Thomcord’ are two of the seedless table transfer/wide somatic hybrids development of grapevines, which were released using this resistant rootstock through gene transfer from technique (Annonymous, 2004). sexually incompatible or difficult to hybridized genera (Grosser et al., 2000). Anther/pollen culture Somatic hybridization research of almost two decades in citrus is yielding fruit now. Application of anther culture is obtaining haploids and homozygous diploids. It In vitro germplasm conservation provides a short cut for obtaining pure lines.As tetraploid plants are produced in In vitro conservation is of great significance greater abundance in breeding programme, it in providing solutions and alternative may be feasible to culture their anther to approaches to overcoming constrains in produced useful diploids. management of genetic resources. Complete plants have been successfully regenerated Regeneration of androgenic embryos in apple from tissues cryopreserved at −196◦C in (Malus x domestica Borkh.) via anther and liquid N2 in several crops for several months microspore culture (Hofer, 2005) has been to years (Ford et al., 2000; Panis et al., 2001; succeeded. Halmagyi et al., 2004; Gupta and Reed, 2006; Ding et al., 2008). This method is now being In vitro mutagenesis practically used at several national and international germplasm banks. Mutagens used physical (gamma rays, X-rays, UV rays) and chemical (EMS, MMS etc.).In In crops, which are propagated vegetatively fruit crops EMS and gamma rays are widely and which produce recalcitrant seeds and used. Notable example of in vitro mutagenesis perennial crops which are highly in Banana variety Grand Nain explants used heterozygous seed storage is not suitable. In was shoot tips; mutagen source and dose were such crops especially, in vitro storage is of 80 Gy gamma rays developed improved great practical importance. These techniques cultivar Novaria-10 with improved trait early have successfully been demonstrated in a flowering and high keeping quality (Mac et number of fruit crops and there are now al., 1996). various germplasm collection centers.

Somatic hybridization A portion of plants regenerated by tissue culture often exhibits phenotypic variation It is crossing of crop plants through fusion of atypical of the original phenotype. Such somatic cells. Protoplast of two cells is fused. variation, termed soma clonal variation may It breaks the barriers of cross incompatibility be heritable i.e. genetically stable and passed among plant species and makes incompatible on to the next generation. Also, presently, crosses possible. Somatic hybridization has synseed technology using encapsulation of been explored for the rootstock and scion non-embryogenic propagules derived under in improvement of citrus. In scion improvement vitro condition has become an important asset production of allotetraploids 2x X 4x = 3x to micropropagation and in vitro conservation (seedless) and direct 3x production whereas, of planting material for long term.

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Somaclonal variation underlies sex differentiation in date palm is not known. Sex of the plants becomes known Somaclonal variation (Larkin and Scowcroft, only at the time of first flowering, which takes 1988) is considered a source of new plant around 5 years. In comparison, molecular genotypes for breeding, and advances at the diagnosis (if available/feasible) promises tissue culture level have opened new quick and reliable identification of sex types possibilities for applications in viticulture. very early when plantlets are growing in The advancements made in tissue culture seedbeds. To develop such an assay, genomic techniques has made it possible to regenerate DNA from 45 individual plants (25 female various horticultural species in vitro as and 20 male) belonging to different varieties micropropagation protocols for commercial of date palm was subjected to PCR scale multiplication are available for a wide amplification using 100 RAPD and 104 ISSR range of crops. However, plant tissue culture (Dhawan et al., 2013). In papaya may generate genetic variability i.e., identification of female and hermaphrodite somaclonal variations as a result of gene specific marker papaya sex determination by mutation or changes in epigenetic marks. The RAPD (Diwedi et al., 2014). occurrence of subtle somaclonal variation is a drawback for both in vitro cloning as well as Genetic diversity assessment germplasm preservation. Somaclonal variation has provided a new and alternative Assessment of diversity has traditionally been tool to the breeders for obtaining genetic made through morphological characters, variability relatively rapidly and without chemical composition, and cytological sophisticated technology in horticultural characters, however, they have several crops, which are either difficult to breed or limitations especially in perennial crops. have narrow genetic base. Somaclonal Morphological characters are often limited in variations are beneficial for creating number, have complex inheritance pattern and variability and resistance to stresses (Table 1). are vulnerable to environmental conditions, hence it is desirable to develop alternative Molecular marker methods, which are rapid, reliable and more or less not influenced by environment. A molecular marker is a DNA sequence that Therefore, in the recent past, rapid use of is readily detected and whose inheritance can molecular markers has complemented the easily be monitored. The phenotype is classical strategies. The use of DNA markers imperfect predictor of genotype. The has enabled the characterization of genotypes phenotype is always expression of its independent of the influences of genotype and environment interaction. Two environmental growth conditions, plants having similar phenotype but genotype physiological age of the plant and the type of point of view they may be different. tissue being analyzed. Molecular markers Application of molecular marker in fruit crops have been looked upon as tools with a large are genetic diversity assessment, estimation of number of applications including localization genetic distance between cultivar and of a gene and improvement of plant varieties phylogenic analysis, detection of quantitative by marker-assisted selection. Molecular trait loci and marker assisted selection. markers thus, are excellent tools for analysis Molecular marker is used in sex of genetic diversity and relationship (Table 2). determination of dioecious plants at juvenile Polymerase chain reaction (PCR) based stage.Genetics of control mechanisms that random amplified polymorphic DNA (RAPD)

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markers have been extensively used in DNA variation in both coding and non-coding finger printing. These DNA markers are region of genome. RAPD analysis is generated by PCR amplification of random technically simple and provides an approach genomic segments with single primers to characterize different genotypes, thus to (usually 10 nucleotides long) of arbitrary estimate genetic diversity which will further sequence (Williams et al., 1990). These be useful in improvement of breeders. markers are mostly dominant and detect

Table.1 Stability of some somaclones to biotic stresses under field condition

Fruit Somaclonal Resistant/tolerant to Field Reference crop variant performance /cultivar Prunus S19-1 and S-156 Bacterial leaf spot *HR after 3 yrs in Hammerschlag persica (Sunhigh) S122- Pseudomonas syringae field et. al., 1994 1 (Redhaven) HR resistance S122-1 compared to Redheaven Apple cv. - Fire blight (Erwinia 60% less Donovan et al., Green amylovora Strain T) symptoms as 1994 Sleeves compared parent cultivar Citrus FS 01 and FS 11 Mal Secco (Phoma Tolerence level Gentile et al., limon tracheiphilla) comparable to 1998 Monachello (T) Banana CIEN BTA-03 Resistance to yellow and Similar Unai et al., 2004; cv. black Sigatoka performance even Gimenez et al., William after 5 year of field 2008 trails

Table.2 DNA markers for genetic diversity assessment in fruit crops

Fruit Marker type Reference 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) Grapes RFLP and SSRs Bourquin et al., (1993) Mango CpSSR and RAPDs He et al., (2007) Marcela et al., (2009) Pistachio Mini satellite marker Riaz Ahmad et al., (2003) Cashew RAPD and ISSR Thimmappaiah et al., (2009) Pear SSRs and AFLP Sisko et al., (2009)

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Table.3 Markers associated to main polygenic traits in fruit crops

Fruit Trait Marker type References

Apple Fire blight resistance SCAR, SSR Sylwia et al., (2009) Citrus Citrus leprosis virus AFLP and RAPD Bastianel et al.,(2009) resistance Pear Incompatibility AFLP and SSR Sun et al., (2009) Banana Sugar content RFLP Ming et al., (2001)

Grapes Seedlessness, Berry AFLP, SSR,RAPDs, Mejía et al., (2007) Size, and Ripening ISSRs and SCARs Date

Strawberry Day-neutrality AFLP Weebadde et al., (2008) Soriano et al., (2007) Apricot Plum Pox Virus SSR

Identification of QTLs background) and Myrobalan plum (green background) on the framework of the Prunus Many important heritable characters are a reference map. Gene abbreviations consequence of the joint action of several correspond to: Y, peach flesh colour; Sharka, genes. Such characters are often referred to as plum pox virus resistance; Mi, nematode polygenic or quantitative. Several characters resistance from peach; D, almond shell of plant species, among which are traits of hardness; Br, broomy plant habit; Dl, double agronomic importance, are inherited flower; Cs, flesh color around the stone; Ag, quantitatively. Yield, maturity date and anther color; Pcp, polycarpel; Fc, flower drought tolerance are examples of such color; Lb, blooming date; F, flesh adherence characters. The genetic loci for such to stone; D, non-acid fruit in peach, Sk, bitter characters have been referred to as kernel; G, fruit skin pubescence; Nl, leaf quantitative trait loci (QTLs). The essential shape; Dw, dwarf plant; Ps, male sterility; Sc, feature which makes feasible the finding and fruit skin color; Gr, leaf colour; Ma, nema- characterization of a QTL is its linkage with a tode resistance from Myrobalan plum; E, leaf known marker locus segregating with gland shape; Sf, resistance to powdery Mendelian ratios. DNA markers provide this mildew. Genes Dl and Br are located on an opportunity by making it feasible to identify, unknown position of G2. map and measure the effects of genes underlying quantitative trait. In grape QTLs Marker assisted selection were used for features such as like critical photoperiod, growth cessation, or dormancy, The use of molecular markers can greatly bud break (BB) and winter hardiness (Table accelerate the pace of selection in fruit crops 3). Approximate positions of 28 major genes with inherent long juvenile period and when were mapped in different populations of trait under study is controlled by the recessive peach (orange background), almond (yellow gene and incorporating more than one gene

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Int.J.Curr.Microbiol.App.Sci (2017) 6(11): 1902-1912 for disease resistance. Presently, the markers regulation of fruit ripening. Using tobacco linked to resistance to Dagger nematode rattle virus-induced gene silencing, the down (Xiphinema index) and pierce disease are regulation of FaSS1 transcripts significantly being routinely used at University of delayed fruit ripening, as evidenced by the California in the breeding programs aimed at changes of firmness, and soluble sugar and improving grapevine rootstock.(Xu et al., anthocyanin contents, as well as the 2008).The resistance to most successful transcripts of several ripening-related genes, resistance gene (Vf) for scab has been including PE1, PL1, XYL2, CHS, CHI and overcome in apple by new race pathogen DFR. Furthermore, the mRNA expression (Gygax et al.,2004). level of FaSS1 was inhibited by abscisic acid or sucrose, but not by glucose after fruit disc Genetic transformation incubation in vitro (Cheng et al., 2017). The two banana E class (SEPALLATA3 [SEP3]) Process of transfer, integration and expression MADS box genes, MaMADS1 and of transgenes in the host cells is known as MaMADS2, homologous to the tomato genetic transformation. Resistant to biotic (Solanum lycopersicum) RIN-MADS ripening stress for example citrus decline, CTV, gene. Transgenic banana plants repressing greening, papaya ring spot virus resistance, either gene (via antisense or RNA plum pox virus etc. Higher tolerance to interference [RNAi]) were created and abiotic stress i.e. salinity, flood, drought etc., exhibited specific ripening delay and shortening the juvenile growth phase and extended shelf-life phenotypes, including quality improvement such as shelf life delayed color development and softening improvement and edible vaccine. (Tomeret al.,2016).

Shelf life improvement Disease resistance

The compositional similarity of the transgenic A viral diseases papaya ring spot virus came papayas with delayed ripening trait with the to Hawaii in the 1940’s and had wiped out non-transgenic control papayas. Fruit papaya production on Oahu by the 1950’s. characteristics such as days to maturity, fruit Transgenicpapaya was developed through weight and total soluble sugars were similar coat protein mediated gene transformation. for both transgenic and control papaya trees The two transgenic varieties were developed (Cabanos, et al., 2014). i.e. Sun Up and Rainbow, those are resistant to Papaya ring spot virus, which Two ACC oxidase genes, CpACO1 and revolutionized the papaya cultivation in CpACO2 are reported and CpACO1 is Hawaii (Gonsalves, 2000). associated with late stage fruit ripening and leaf senescence whereas CpACO2 is induced Plum pox virus causes fruit deformation and before colors break (Chen et al., 2003). Two reduced quality, premature fruit drop, leaf of the ripening-related genes MaMads-rin and chlorosis, tree decline and death in severe MaExp2 are used for banana transformation infections or co-infections with other viruses. in order to increase shelf-life and fruit quality Honey Sweet an improved cultivar of plum Liu et al., (2009). Juan et al., (2009) have was developed using mechanism of RNAi shown that PG plays a significant role in silencing, which was resistant to virus strawberry ripening. Two ripening-related PG (Ravelonandro et al., 2000). genes, FaPG1 and FaPG2 identified for

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Future prospect Bhat, Z.A., Dhillon W.S., Rashid R., Bhat, J.A., Dar W.A., Ganaie M.Y (2010). Plant biotechnology has the potential to play a The role of Molecular Markers in key role in the sustainable production of fruit Improvement of Fruit Crops. Not Sci crops. However, there is enormous potential Biol 2 (2) 22-30. for genetic manipulation of some vegetative Botez, C., R. Sestras, M. Ardelean, D. Pamfil, propagated fruit crops in order to improve B. Patrascu and A. Sestras (2009). their disease and pest resistance. The use of Phenotypic Selection Assisted by appropriate constructs may allow the Molecular Markers for Scab Resistance production of nematode, fungal, bacterial and in Apple. Acta Hort. 814:771-776. virus resistant plants in a significantly shorter Botstein, J. O., A. Sonko, L. Otten and B. period of time than using conventional Walter (1993). RFLP molecular breeding, especially if several traits can be taxonomy in vinifera L. Theor. introduced simultaneously. It may also be Appl. Genet. 87:431-38. possible to incorporate other characteristics Bourquin, J. C., A. Sonko, L. Otten and B. such as drought tolerance, thereby extending Walter (1993). RFLP molecular the geographic spread of some fruit crops for taxonomy in . Theor. and production, and thus contributing Appl. Genet. 87:431-38. substantially to enhanced food security and Cerrone, S. C., Andres, G. S., Roberta, N. G., poverty alleviation. However, high caution is Simeona, V. S. and Evelyn, M. T. M. required for biosafety experiments and 2003. Compositional analysis of potential risk assessment bearing in mind that transgenic papaya with delayed ripening these are consumed by most humans and trait. Philippines Agricultural Scientist, mainly by children. 96(4): 331–339 Chen Y.T., Lee, Y.R., Yang, C.Y., Wang, Y. References T., Yang, S. F., Shaw, J. F. 2003. A novel papaya ACC oxidase gene (CP- Anonymous (2004). FPS Grape Program ACO2) is associated with late stage fruit Newsletter October, 2004. Foundation ripening and leaf senescence. Plant Plant Services, pp:4. Science, 164:531–40. Ashworth, V. E. and M. T. Clegg (2003). Cheng Zhao, Li-Na Hua, Xiao-Feng Liu,Yu- Microsatellite Markers in Avocado Zhong Li, Yuan-Yue Shen, Jia-Xuan (Prunus americana Mill.): Genealogical Guo.2017.Sucrose synthase FaSS1 Relationships Among Cultivated plays an important role in the regulation Avocado Genotypes Journal of of strawberry fruit ripening. Plant Heredity 94:407-415. Growth Regulaters, 81:175–181. Bastianel, M., L. Cristofani-Yaly, A. C. M. Coart, E., X. Vekemans, M. J. M. Smulders, I. Oliveira, J. Freitas-Astúa, A. Antonio, Wagner, J. Van Huylenbroeck, E. Van G. M. Franco, D. V. Resende, V. Bockstaele and I. Roldán-Ruiz (2003). Rodrigues and M. A. Machado (2009). Genetic variation in the endangered Quantitative trait loci analysis of citrus wild apple (Malussylvestris L. Mill.) in leprosis resistance in an interspecific Belgium as revealed by amplified backcross family of (Citrus reticulata fragment length polymorphism and Blanco × C. sinensis L. Osbeck) × C. microsatellite markers. Mol. Ecology sinensis L. Osb. Euphytica 169:101- 12:845-857. 111. Dhawan, C., Kharb, P. and Sharma R. (2013).

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How to cite this article:

Anuradha, Subhash Chander and Arvind Malik. 2017. Biotechnology a Modern Tool for Fruits Production - A Review. Int.J.Curr.Microbiol.App.Sci. 6(11): 1902-1912. doi: https://doi.org/10.20546/ijcmas.2017.611.226

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