Int. J. Biosci.13(2), 67-75, August 2018
Cowpea is one of the most important protein rich legumes worldwide. This study used the Random Amplified Polymorphic DNA (RAPD) assays to reveal the positive and negative DNA markers in seven cultivars of cowpea, which could use for cultivars identification and discrimination, for future breeding programs and derivation of novel genotypes. Also, to evaluate the proportion of genetic relationship among the cultivars by detecting the genetic polymorphism at molecular level and correlate the obtained results with some morphological traits and source of cultivars. RAPD Results showed that a total number of 56 DNA fragments were amplified, ranged in size between 1637 bp and 163 bp, while the total polymorphic bands number was 45 and the percentage of polymorphism was 80.3 %. Five cultivars (Cream-12, Chinese red, Blackeye crowder, Brown crowder, and Cream-7) appeared to have whether positive or negative markers with different molecular weight, while Azmerly and Dokki-331 cultivars did not produce any specific bands. The dendrogram of RAPD data showed that the 7 cultivars of cowpea were grouped in 6 clusters based on genetic similarities; the highest value of similarity was 86.6 % between Azmerly and Dokki-331, while the lowest was between Cream-12 and Blackeye crowder with a value of 54.2 %. The obtained results suggest that RAPD markers were better linked to the source of the cultivar and to the extent of seed crowding in pod trait, while the seed colour and the growth habit traits might or might not show correlation with the molecular data.
Ajibade SR, Weeden NF, Chite SM. 2000. Inter simple sequence repeat analysis of genetic relationships in the genus Vigna. Euphytica 111, 47–55. http://dx.doi.org/10.1023/A:1003763328768.
Ali ZB, Yao KN, Odeny DA, Kyalo M, Skilton R, Eltahir IM. 2015. Assessing the genetic diversity of cowpea (Vigna unguiculata (L.) Walp.) accessions from Sudan using simple sequence repeat (SSR) markers. African Journal of Plant Science 9, 293–304. http://dx.doi.org/10.5897/AJPS2015.1313.
Ba FS, Pasquet RS, Gepts P. 2004. Genetic diversity in cowpea (Vigna unguiculata (L.) Walp.) as revealed by RAPD markers. Genetic Resources and Crop Evolution 51, 539–550. http://dx.doi.org/10.1023/B:GRES.0000024158.83190.4e.
Damarany AM. 1994. Testing and screening of some cowpea (Vigna unguiculata(L.) Walp.) genotypes under Assiut conditions. Assiut Journal of Agricultural Sciences 25, 9-19.
Diouf D, Hilu KW. 2005. Microsatellites and RAPD Markers to Study Genetic Relationships Among Cowpea Breeding Lines and Local Varieties in Senegal. Genetic Resources and Crop Evolution 52, 1057–1067. http://dx.doi.org/10.1007/s10722-004-6107-z.
FAO. 2016. Food and Agriculture Organization of the United Nations. Cowpea FAOSTAT data. http://www.fao.org/faostat.
Ghalmi N, Malice M, Jacquemin JM, Ounane SM, Mekliche L, Baudoin JP. 2010. Morphological and molecular diversity within Algerian cowpea (Vigna unguiculata (L.) Walp.) landraces. Genetic resources and crop evolution 57, 371-386. http://dx.doi.org/10.1007/s10722-009-9476-5.
Khan T, Reza OH, Khan A, Haque S, Islam S, Badiuzzaman K. 2015. Genetic Diversity Analysis of Cowpea by RAPD Markers. International Journal of Innovation and Applied Studies 10, 459–465.
Kumar NS, Gurusubramanian G. 2011. Random amplified polymorphic DNA (RAPD) markers and its applications. Science Vision 11, 116-124. http://dx.doi.org/10.1.1.404.1046.
Kumari N, Thakur SK. 2014. Randomly Amplified Polymorphic Dna-A Brief Review. American Journal of Animal and Veterinary Sciences 9, 6–13. http://dx.doi.org/10.3844/ajavssp.2014.6.13.
Murray MG, Thompson W. 1980. Rapid isolation of high molecular weight plant DNA. Nucleic acids research 8, 4321-4326.
Nagalakshmi RM, Kavitha MJ, Ananda KCR, Usha KR. 2017. Genetic Diversity in Cowpea (Vigna unguiculata (L.) Walp.), Using RAPD Markers. Journal of Pharmacognosy and Phytochemistry 6, 1632-1635.
Nei M, Li WH. 1979. Mathematical model for studying genetic variation in terms of restriction endonucleases. Proceedings of the National Academy of Sciences 76, 5269-5273.
Obiadalla-Ali HA, Mohamed NEM, Khaled AGA. 2015. Inbreeding, outbreeding and RAPD markers studies of faba bean (Viciafaba L.) crop. Journal of Advanced Research 6, 859–868. http://dx.doi.org/10.1016/j.jare.2014.07.004.
Ouédraogo JT, Gowda BS, Jean M, Close TJ, Ehlers JD, Hall AE, Gillaspie AG, Roberts PA, Ismail AM, Bruening G, Gepts P, Timko MP, Belzile FJ. 2002. An improved genetic linkage map for cowpea (Vigna unguiculata L.) Combining AFLP, RFLP, RAPD, biochemical markers, and biological resistance traits. Genome 45, 175–188. http://dx.doi.org/10.1139/g01-102.
Saghai-Maroof MA, Soliman KM, Jorgensen RA, Allard RWL. 1984. Ribosomal DNA spacer-length polymorphisms in barley: Mendelian inheritance, chromosomal location, and population dynamics. Proceedings of the National Academy of Sciences 81, 8014-8018. http://dx.doi.org/10.1073/pnas.81.24.8014.
Saker MM, Youssef SS, Abdallah NA, Bashandy HS, El-Sharkawy AM. 2005. Genetic analysis of some Egyptian rice genotypes using RAPD, SSR and AFLP. African Journal of Biotechnology 4, 882-890. http://dx.doi.org/10.4314/ajb.v4i9.71227.
Udensi OU, Okon EA, Ikpeme EV, Onung OO, Ogban FU. 2016. Assessing the Genetic Diversity in Cowpea (Vigna unguiculata L. Walp) Accessions Obtained from IITA, Nigeria Using Random Amplified Polymorphic DNA (RAPD). International Journal of Plant Breeding and Genetics 10, 12–22. http://dx.doi.org/10.3923/ijpbg.2016.12.22.
Williams JG, Kubelik AR, Livak KJ, Rafalski JA, Tingey SV. 1990. DNA polymorphisms amplified by arbitrary primers are useful as genetic markers. Nucleic acids research 18, 6531-6535. http://dx.doi.org/10.1093/nar/18.22.6531.