Development of tetraploid plants from an interspecific hybrid between mungbean ( Vigna radiata ) and rice bean ( Vigna u
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J. Crop Sci. Biotech. 2013 (March) 16 (1) :45~51 DOI No. 10.1007/s12892-012-0078-y RESEARCH ARTICLE
Development of Tetraploid Plants from an Interspecific Hybrid between Mungbean (Vigna radiata) and Rice Bean (Vigna umbellata) Tanapon Chaisan1,2, Prakit Somta3, Peerasak Srinives3*, Sontichai Chanprame1,2,3, Rangsarid Kaveeta4, Surapong Dumrongkittikule5 Center for Agricultural Biotechnology, Kasetsart University, Kamphaeng Saen, Nakhon Pathom 73140, Thailand Center of Excellence on Agricultural Biotechnology (AG-BIO/PERDO-CHE), Bangkok 10900, Thailand 3 Department of Agronomy, Faculty of Agriculture at Kamphaeng Saen, Kasetsart University, Kamphaeng Saen, Nakhon Pathom 73140, Thailand 4 Department of Agronomy, Faculty of Agriculture, Kasetsart University, Chatuchak, Bangkok 10900, Thailand 5 Department of Horticulture, Faculty of Agriculture at Kamphaeng Saen, Kasetsart University, Kamphaeng Saen, Nakhon Pathom 73140, Thailand
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Received: July 29, 2012 / Revised: November 17, 2012 / Accepted: January 12, 2013 Ⓒ Korean Society of Crop Science and Springer 2013
Abstract Mungbean (Vigna radiata) and rice bean (V. umbellata) (both species 2n = 2x = 22) have desirable traits that complement each other. In this study, we rescued embryos from a cross between mungbean cv. “Kamphaeng Saen 2” and rice bean cv. “Miyazaki” and resolved the hybrid sterility problem by colchicine treatment. The interspecific hybrids were obtained when Kamphaeng Saen 2 was used as the female parent. Four out of 80 immature seeds at 12 days old were able to germinate on an MS medium supplemented with 1 mg L-1 IAA, 0.2 mg L-1 kinetin, and 500 mg L-1 casein hydrolysate. Forty random amplified polymorphic DNA (RAPD) primers were screened for polymorphism among the parents, and two specific primers were finally chosen for testing of hybridity. Using the two primers, all putative F1 hybrids were confirmed as the interspecific hybrids. To observe their fertility, some of the hybrid seedlings were transplanted. The hybrid produced flowers profusely but failed to set pods. To overcome the sterility, plants were induced to become tetraploid by colchicine treatment in vitro. The ploidy level of the regenerated seedlings was confirmed from leaf DNA using a flow cytometer. Three out of 20 hybrid seedlings (15%) were successfully induced from diploid to tetraploid by a colchicine concentration of 2 g L-1. The tetraploid hybrids were able to produce flowers and set pods normally. Key words: embryo rescue, interspecific hybridization, in vitro tetraploid induction, Vigna radiata, Vigna umbellata Abbreviations: 6-BAP: 6-benzylaminopurine, BK medium: Brewbaker and Kwak medium, DAH: days after hybridization, FCM: flow cytometry, IAA: Indole-3-acetic acid, MS medium: Murashige and Skoog medium, RAPD: random amplified polymorphic DNA, TBE buffer: tris-borate-EDTA buffer
Introduction Mungbean (Vigna radiata (L.) Wilczek) (2n = 2x = 22) is the most widely-grown Vigna species in Thailand. The crop has long been domesticated and thus exists in a more desirable ph
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