Appliance Of Rapd Markers In The Analysis Of Genetic  Variations In Five Mango Cultivars

Ramadasappa Pratibha; Mallaiah Shivashank

doi:10.48165/

Authors

Ramadasappa Pratibha Department of Biotechnology and Genetics, Ramaiah College of Arts, Science and Commerce, MSRIT Post, Bengaluru – 560 054, Karnataka (India)
Mallaiah Shivashank Department of Life Sciences, Jnana Bharathi Campus, Bangalore University, Bengaluru - 560 056, Karnataka (India)

DOI:

https://doi.org/10.48165/

Keywords:

DNA, mango cultivars, PCR, phylogenetic tree, RAPD

Abstract

The aim of this work was to analyze molecular relationship between five mango (Mangifera indica) cultivars viz., ‘Alphanso’, ‘Totapuri’, ‘Langra’, ‘Kesar’ and ‘Amrapalli’ using molecular RAPD markers. In present study, genomic DNA isolation procedure was standardized to extract DNA from mango pulp. All the samples tested yielded good quality DNA as 260/280 nm ratio was within 1.6-1.8 range revealing that no significant amount of proteins was present in extract. The statistical analysis (ANOVA) indicated p ≤0.05 validating the applicability of optimized protocol. The extracted DNA was subjected to PCR with 39 primers. Thirty six RAPD primers produced distinct and consistent RAPD profiles which revealed 160 bands with decent 87% polymorphism. Neighbour joining tree, constructed based on Nei’s genetic distance, displayed splitting of tested cultivars into two separate clades designated as I and II. The method employed in this study highlighted the reproducibility of technique and, therefore, appears to be a useful tool for genetic analysis in mango and other related plant species.

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References

Anonymous. 2006. Area and Production of Horticultural Crops. Bulletin of National Horticulture Board. National Horticulture Board, Bangalore, India.

Arús, P., Yamamoto, T., Dirlewanger, E. and Abbott, A.G. 2006.Synteny in the Rosaceae. Plant Breeding Reviews, 27: 175.

Collins, D., Mill, R.R.and Möller, M. 2003. Species separation of Taxusbaccata, T. canadensis and T. cuspidata (Taxaceae) and origins of their reputed hybrids inferred from RAPD and cpDNA data. American Journal of Botany, 90: 175-182.

Das, A., Spackman, E., Pantin-Jackwood, M.J. and Suarez, D.L. 2009. Removal of real-time reverse transcription polymerase chain reaction (RT-PCR) inhibitors associated with cloacal swab samples and tissues for improved diagnosis of Avian influenza virus by RT-PCR. Journal of Veterinary Diagnostic Investigation, 21: 771-778.

Debener, T., Mattiesch, L. and Vosman, B. 2000. A molecular marker map for roses. pp. 283-287. In: IV International Symposium on Rose Research and Cultivation. International Society for Horticultural Science, Hannover, Germany.

Fabbri, A., Hormaza, J.I. and Polito, V.S. 1995. Random amplified polymorphic DNA analysis of olive (Olea europaea L.) cultivars. Journal of the American Society for Horticultural Science, 120: 538-542.

Kaemmer, D., Afza, R., Weising, K., Kahl, G. and Novak, F.J. 1992. Oligonucleotide and amplification fingerprinting of wild species and cultivars of banana (Musa spp.). Nature Biotechnology, 10: 1030-1037.

Use of RAPD markers in genetic analysis mango cultivars 205

Kimura, M., and Weiss, G.H. 1964. The stepping stone model of population structure and the decrease of genetic correlation with distance. Genetics, 49: 561-569.

Koller, B., Lehmann, A., McDermott, J.M. and Gessler, C. 1993. Identification of apple cultivars using RAPD markers. Theoretical and Applied Genetics, 85: 901-904.

Kumar, S., Stecher, G. and Tamura, K. 2016. MEGA7: Molecular evolutionary genetics analysis version 7.0 for bigger datasets. Molecular Biology and Evolution, 33: 1870-1874. Lewontin, R.C. 1972. The apportionment of human diversity. pp. 381-398. In: Evolutionary Biology. Springer, New York, USA.

Moyo, M., Amoo, S.O., Bairu, M.W., Finnie, J.F. and Van Staden, J. 2008. Optimizing DNA isolation for medicinal plants. South African Journal of Botany, 74: 771-775. Naidu, G.M. and Naidu, G.R. 2009. Marketing strategies for exporting mangoes and mango products from India. Acta Horticulturae, 820: 79-95.

Negi, S.S. 1999. Mango production in India. pp. 69-78. In: VIth International Symposium on Mango. 6th April, 1999. Central Institute for Subtropical Horticulture, Lucknow, India. NHB. 2018. Indian Horticulture Database. National Horticulture Board, Ministry of Agriculture, Government of India, Gurgaon, Haryana, India [www.agriexchange.apeda.gov.in]. Padmalatha, K. and Prasad, M.N.V. 2006. Optimization of DNA isolation and PCR protocol for RAPD analysis of selected medicinal and aromatic plants of conservation concern from Peninsular India. African Journal of Biotechnology, 5: 230-234.

Pathirana, C.K., Chandrasekara, C.H.W.M.R.B., Attanayake, S.R.M.R., Weerakkody, W.A.P., Eeswara, J.P. and Bandaranayake, P.C.G. 2018. Optimization of DNA extraction and PCR protocols for plants with high Phenolics: Bael, mango, pomegranate as examples. Ceylon Journal of Science, 47: 43-48.

Pavlicek, A., Hrda, S. and Flegr, J. 1999. Free-tree-software program for construction of phylogenetic trees on the basis of distance data and bootstrap/jackknife analysis of the tree robustness. Application in the RAPD analysis of genus Frenkelia. Folia Biologica (Praha), 45: 97-99.

Porbeski, S., Bailey, L.G. and Baum, B.R. 1997. Modification of a CTAB DNA extraction protocol for plants containing high polysaccharide and polyphenol components. Plant Molecular Biology Reporter, 15: 8-15.

Pratibha, R. and Shivashankar, M. 2018. Comparative analysis on the quality characteristics of commercially important mango cultivars for canning. International Journal of Fundamental and Applied Sciences Comparative, 7: 13-17.

Pruthvish, R. and Chikkaswamy, B.K. 2016. Genetic diversity and relationships among mango cultivars using RAPD molecular markers. International Journal of Current Microbiology and Applied Sciences, 5: 778-787.

Ramakrishnaiah, T.N., Sreeramulu, A. and Ramakrishna, S. 2018. Morphometric analysis of oxyopid spiders (Araneae: Oxyopidae) from Karnataka. Indian Journal of Entomology, 80: 211-216.

Roldàn-Ruiz, I., Dendauw, J., Van Bockstaele, E., Depicker, A. and De Loose, M.A.F.L.P. 2000. AFLP markers reveal high polymorphic rates in ryegrasses (Lolium spp.). Molecular Breeding, 6: 125-134.

Saitou, N. and Imanishi, T. 1989. Relative efficiencies of the Fietch-Margoliash, maximum parsimony, maximum-likelihood, minimum-evolution and Neighbor-joining methods of phylogenetic tree construction in obtaining the correct tree. Molecular Biology and Evolution, 6: 514-514.

Sambrook, J. and Russell, D.W. 2001. Molecular Cloning: A Laboratory Manual Volume 1 (3rd edn.). Cold Spring Harbour Laboratory, Cold Spring Harbour, New York, USA. Schnell, R.J., Ronning, C.M. and Knight, R.J. 1995. Identification of cultivars and validation of geo-magnetic relationships in Mangifera indica L. using RAPD markers. Theoretical and Applied Genetics, 90: 269-274.

Ramadasappa Pratibha and Mallaiah Shivashank

Schubert, J., Habekuß, A. and Rabenstein, F. 2003. Investigation of differences between wheat and barley forms of wheat dwarf virus and their distribution in host plants. Plant Protection Science, Prague, 38: 43-48.

Shahzadi, I., Ahmed, R., Hassan, A. and Shah, M.M. 2010. Optimization of DNA extraction from seeds and fresh leaf tissues of wild marigold (Tagetes minuta) for polymerase chain reaction analysis. Genetics and Molecular Research, 9: 386-393.

Shivashankar, M. 2015. Study on RAPD molecular distinction in mulberry cultivars. International Journal of Current Microbiology and Applied Sciences, 4: 1097-1105.

Tajima, F. 1989. Statistical method for testing the neutral mutation hypothesis by DNA polymorphism. Genetics, 123: 585-595.

Welsh, J. and McClelland, M. 1990. Fingerprinting genomes using PCR with arbitrary primers. Nucleic Acids Research, 18: 7213-7218.

Williams, J.G., Kubelik, A.R., Livak, K.J., Rafalski, J.A. and Tingey, S.V. 1990. DNA polymorphisms amplified by arbitrary primers are useful as genetic markers. Nucleic Acids Research, 18: 6531-6535.

Yeh, F.C., Yang, R.C. and Boyle, T. 1999. Popgene Version 1.31. Microsoft Window-based Freeware for Population Genetic Analysis. University of Alberta, Edmonton, Canada.