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Genetic Diversity and Relationships Among Mango Varieties Using RAPD Molecular Markers

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Genetic Diversity and Relationships Among Mango Varieties Using RAPD Molecular Markers Int.J.Curr.Microbiol.App.Sci (2016) 5(1): 778-787 ISSN: 2319-7706 Volume 5 Number 1(2016) pp. 778-787 Journal homepage: http://www.ijcmas.com Original Research Article http://dx.doi.org/10.20546/ijcmas.2016.501.079 Genetic Diversity and Relationships among Mango Varieties using RAPD Molecular Markers R. Pruthvish1 and B.K.Chikkaswamy2* 1Department of Biotechnology, Acharya Institute of Technology, Soladevanahalli Hesaraghatta Road, Bangalore, India 2Sigma BioScience Research Center, Indira Nagar Bangalore-560038, India *Corresponding author A B S T R A C T K e y w o r d s Mango is the genus of tree & bushy tree in the family Mangnoliaceae. Most of the Genetic Mango varieties are dioceous and cross-pollinate to produce to produce fertile Diversity, hybrids suggesting a closer genetic relationship which is not expected at species Mango level. The traditional methods using morphological characters are not successful in Varieties, establishing the diversity and relationship among 19 different Mango varieties RAPD, because of the environmental influence.PCR based molecular marker method, Molecular RAPD was employed to study the genetic diversity and Inter-relationship among Markers 19 Mango varieties. On an average RAPD analysis generated 5-6 discrete bands/ varieties with 10 nucleotides primers. The size of the amplified products ranged Article Info from 100-3500 base pairs in length. With an average of 5-10 bands per primer. Of Accepted: 21 amplified fragments 50 were polymorphic (80%) with at least one pair wise 25 December 2015 comparison between 19 varieties. RAPD analysis identified varieties specific Available Online: amplification products which will be useful in germplasm classification and 10 January 2016 introgression studies. These results indicated that RAPD based markers are useful for genetic characterization of Mango /varieties/accessions. Introduction Genetic fingerprinting has been expense, and use of (32 P) for labeling accomplished traditionally through the use with RELPs and VNTRs. Polymerase Chain of isozymes, and more recently through Reaction (RAPD) uses arbitrary 10-base reactions fragment length polymorphisms primers to amplify randow portions of the (RELPs), variable number tandem repeats genome (Welsh and McCIelland 1990; (VNTRs), or a combination of these. While Williams et al. 1990).The fragments these methods have been very useful in produced are easily visualized on an cultivar identification, they have a number ethidium bromide stained gel, and of disadvantages, including a limited polymorphisms between genotypes reflect number of isozyme loci, and the time, heritable differences in the genome. Large 778 Int.J.Curr.Microbiol.App.Sci (2016) 5(1): 778-787 numbers of bands, or loci, can be generated Until recently, several promising species with relative ease. Due to the arbitrary released formally are characterized on the nature of the primers, RAPD markers, unlike basis of morphological data, mango content RELPs, VNTRs, and isozymes, represent a and yield potential. Worldwide demands on random sample from the entire genome. mango necessitate work on conservation of They are, however, inherited as dominant mango germplasm and their further genetic markers, requiring larger numbers of loci to improvement. Therefore precise be identified and screened to glean the same characterization of promising species and information as from RELPLs, VNTRs, and determination of genetic variation among isozymes. Amplification of bands in the those are felt necessary. progeny which were not amplified in either parent has also been reported (Riedy et al. Until recently, several promising species 1992). released formally are characterized on the basis of morphological data, mango content The use of RAPDs to determine genetic and yield potential. These characters differ relationships has been demonstrated in under varying environmental condition maize (Welsh et al. 1991), conifers (Carlson thereby posing problem identification of et al. 1991), caca (Wide et al. 1992), and species. Unlike morphological markers, various leguminous pecies (Charlmers et al. cytological (chromosome numbers, nuclear 1992, Echt et, al.1992). Within Mangifera DNA content) and molecular markers (mango) species RAPDs have been used to (RAPD, AFLP, ISSR etc.) are not prone to determine phylogenetic relationships environmental influences and characterize (Schnell and Knight 1993). On the basis of the plants portraying the extent of genetic the classification of Kostermans and Bom. diversity among taxa (Bennett 1987,Bennett and Smith 1991,Waugh and Powell RAPD fingerprinting techniques have been 1992,Chalmers et al.1994, Das et al.1998, used for the identification of horticultural Rodriguez et al.1999, Das et al.2001). crop varieties, description of cultivars genotypes and for protecting breeder s rights Of the different markers, RAPD has been (Williams et al., 1990) enabling assay to be widely used in the last decade in species performed at any stage of plant identification programmed (Schnell et development. RAPD markers have been al.1995) and in assessing genetic variations extensively used to distinguish intraspecific among different taxa at DNA level because genetic variation in ornamental crops and of its cost effectiveness and simple operation detection of hybrids and clones (Collins et without requiring prior knowledge of al., 2003; Arus, 2000; Debener, 2001a). species DNA sequences (Williams et al.1990, Frankel et al.1995). RAPDs reveal Ranamukhaarachchi et al. (2001) showed similar patterns of genetic diversity when that RAPD markers had the ability to compared with other marker types and can identify pot-plant mango cultivars is crucial be performed more rapidly than most other to local breeders involved in hybridization methods (Morell et al.1995) and can provide programs for varietal development but is vital information for the development of currently lacking. In this study, RAPD genetic sampling, conservation and markers were used to evaluate the extent of improvement strategies (Waugh and Powell genetic variation among some mango cut 1992, Chalmers et al.1994). No report has flower cultivars. been published so far either on the genetic 779 Int.J.Curr.Microbiol.App.Sci (2016) 5(1): 778-787 characterization or on the extent of genetic out on RAPD molecular marker studies in variations existing among promising species varieties of mango plants with the following of mango. objectives. To develop protocol for isolation of DNA from different 19 varieties of The present study deals with the in situ mango plants. To develop PCR protocol and DNA estimation and RAPD analysis of 19 to identify RAPD markers to score in promising species of Mango to identify and various mango plants.To assess the genetic evaluate extent of genetic variation existing diversity and relationships among mango among these. plants using RAPD molecular markers belonging to family magnoliace. As the efficiency of selection scheme or genetic analysis based on phenotype is a Matias Kirst et.al.,(2004) studied on DNA function of heritability of the trait, factors marker RAPD,RFLPs and AFLP etc in like environment, traits of mutagenic and forest tree species and in particular its quantitative inheritance or partial and application to tree breeding and tree complete dominance often confound the genomics. expression of genetic traits. Many of these complications of a phenotype-based assay Million and Chinnappa (2000) assessed the can be overcome through direct genetic divergence in forty seven genotypes identification of species with DNA based of diagnostic assay. For this reason DNA based genetic markers are being integrated into Stellaria longipes (Caryophyllaceae) using several plant systems under expected to play RAPD analysis. Datta and Mitrick an important role in the future plant et.al.,(1997) studied the classification of improvement programmes. common bean Phaseolus vulgaris from Chile using RAPD data.The study reveals Advent of Polymerase chain reaction (PCR) that 95 bean accessions analysed using 25 technology as lead to the development of primers that generated 106 polymorphic several novel genetic assays based on bands of RAPD collected from different selective DNA amplification Krawetz locations. (1989) and Innis et al (1990). RAPD assay detects nucleotides sequence of Nirupama et.al., (2003). Reported genetic polymorphism in DNA using only a single diversity and relationship of 51 accessions primer of arbitrary nucleotide sequence. The of Vetiver using RAPD/AFLP analysis.Total protocol is also relatively quick and easy to of 20 primers were used to generate 5 perform and uses fluorescence instead of clusters using UPGMA cluster analysis radioactivity. Because the RAPD technique where as for AFLP analysis nine primers are is amplification- based assay, only used to generate 383 unique bands specific nanogram quantity of DNA is required. One to different accessions compared 81 of the strengths of these new assays is that monomorphic bands.In contrast sufficient they are more amenable to automation than diversity existing with in the wild and conventional techniques. It is simple to cultivated.Indian germplasm. Raghvendra perform and is preferable to experiment; saxena and Amaresh Sanghamitra Nayak their species of large number of individuals et.al. (2006) studied the 4C nuclear DNA are to be determined at a few genetic content and RAPD analysis of 17 promising loci.The present investigation will be carried cultivars of turmeric Curcuma longa of 780 Int.J.Curr.Microbiol.App.Sci
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