CIBTech Journal of Biotechnology ISSN: 2319–3859 (Online) An Open Access, Online International Journal Available at http://www.cibtech.org/cjb.htm 2014 Vol. 3 (4) October-December, pp.11-15/Himabindu et al. Research Article © Copyright 2014 | Centre for Info Bio Technology (CIBTech) 11 IN VITRO REGENERATION OF GREEN GRAM [VIGNA RADIATA (L.) WILCZEK] CULTIVAR VAMBAN-2 USING COTYLEDONARY NODES Himabindu Y. 1, 2 , Madhava C Reddy 2 and *Chandrasekhar T. 1 1 Department of Environmental Science, Yogi Vemana University, Kadapa-516003, Andhra Pradesh, India 2 Department of Biotechnology & Bioinformatics, Yogi Vemana University, Kadapa-516003, Andhra Pradesh, India *Author for Correspondence ABSTRACT Efficient protocols have been developed to induce shoot multiplication from cotyledonary node cultures of green gram [Vigna radiata (L.) Wilczek] cultivar Vamban-2. Cotyledonary nodal segments cultured on Gamborg’s medium (B5) containing 0.5 mg/l 6-benzyladenine (BA) induced on an average of nine multiple shoots. The effect of agar on shoot multiplication was studied and we did not found much difference between the treatments. Regenerated shoots showed 65 % rooting on full strength B5 basal medium without any plant growth regulators. In vitro regenerated plantlets were successfully acclimatized to greenhouse conditions. Keywords: Green Gram, Vamban, Agar, Cotyledonary Node, B5 Medium INTRODUCTION Green gram or mung bean [Vigna radiata(L.) Wilczek] is one of the most important pulse crops in India and cultivated in different parts of the world. Protein rich edible seeds, sprouts rich in vitamins and amino acids are used directly and apart from this the crop is widely used as forage (Kaviraj et al., 2006). However, the productivity and quality of the grain is severely reduced due to different stress factors in many parts of the country. Despite its great economic importance little information regarding its degree of stress tolerance is available through conventional studies, although yield losses are considerable when subjected to different stress conditions. But using genetic engineering methods we can achieve any kind of stress tolerance by producing transgenic green gram plants using different resistant genes (Yadav et al., 2012). But before that the establishment of in vitro regeneration protocol is prerequisite for genetic transformation methods. Micropropagation is an alternative to the conventional method of vegetative propagation with the objective of enhancing the rate of multiplication. Most of the green gram in vitro micropropagation focuses on different explants including cotyledonary node multiplication, which is ideal for high clonal fidelity and efficiency (Khatun et al., 2008). Protocols have been established for multiplication from different green gram cultivars using cotyledonary node explants (Gulati et al., 1994; Janaki and Manoharan, 2012; Vats et al., 2014). However existing regeneration protocols for green gram are not repeatable across different species, i.e. are to a significant extent species-specific. There are a limited number of laboratories running mass production of green gram at commercial scale due to technical and commercial reasons.Keeping in view the importance of clonal propagation through cotyledonary node multiplication and lack of information on this protocol with Vamban-2 species, we are interested in establishing a highly reproducible protocol for large-scale micropropagation of this cultivar. MATERIALS AND METHODS Healthy and uniform seeds of mung bean cultivar Vamban-2 were rinsed with distilled water for 5-6 times and then with 70% ethanol for 1 min. The seeds were then surface sterilized with 0.1% HgCl2 for 8-10 min followed by four to five times repeated washings with sterile distilled water. The surface sterilized seeds were asceptically germinated on water agar medium. Cotyledonary node segments from 2-3 day old seedlings were cultured on shoot multiplication medium.