American Journal of Plant Sciences, 2013, 4, 2001-2005 http://dx.doi.org/10.4236/ajps.2013.410249 Published Online October 2013 (http://www.scirp.org/journal/ajps) 2001 In Vitro Plant Regeneration of Morus indica L. cv. V1 Using Leaf Explant Melur Kodandaram Raghunath 1 , Karaba N. Nataraja 2* , Jyothi Sainath Meghana 1 , Radha Sivarajan Sajeevan 2 , Mala V. Rajan 1 , Syed Mashayak Hussaine Qadri 1 1 Central Sericultural Research and Training Institute, Srirampura, Mysore, India; 2 Department of Crop Physiology, University of Agricultural Sciences, GKVK, Bangalore, India. Email: * nataraja_karaba@yahoo.com Received March 26 th , 2013; revised April 27 th , 2013; accepted May 19 th , 2013 Copyright © 2013 Melur Kodandaram Raghunath et al. This is an open access article distributed under the Creative Commons Attribu- tion License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. ABSTRACT Adventitious bud induction and plantlet regeneration were studied in a popular mulberry variety, V1 using leaf as an explant. Fully expanded leaf explants were cultured on Murashige and Skoog’s (MS) medium supplemented with thidi- azuron (TDZ) (0.5 - 4.0 mg/l), 6-benzylaminopurine (BAP) (0.5 - 2.0 mg/l), indole acetic acid (IAA) (2.0 mg/l), gibber- lic acid (GA 3 ) (1.0 - 2.0 mg/l) silver nitrate (AgNO 3 ) (2.0 mg/l) and different carbon sources such as sucrose, fructose and glucose (10% - 30%) either individually or in combination to induce adventitious buds and regeneration. The high- est percentage (63%) of adventitious bud formation and regeneration (68%) was achieved in the medium containing MS with TDZ (1.0 mg/l), IAA (2.0 mg/l) and AgNO 3 (2.0 mg/l). For subsequent regeneration and shoot elongation the MS medium having BAP (1.0 mg/l), GA3 (2.0 mg/l) and AgNO 3 (2.0 mg/l) was found to be suitable. Amongst the carbon sources tested, the most suitable carbon source was found to be sucrose (3%) followed by fructose (2%) for adventitious bud formation. Excised in vitro shoots were rooted (60% - 80%) in half strength MS medium supplemented with in- dole-3-butyric acid (1.0 mg/l). The well rooted plantlets were hardened in soil + sand + farm yard manure (FYM) mix- ture with a success rate of 70% - 90%. Since in vitro regeneration is highly genotype-dependent in mulberry, the stan- dardized protocol can be effectively used for further improvement of this leading genotype using biotechnological ap- proaches. Keywords: In Vitro Regeneration; Adventitious Bud; Thidiazuron; Mulberry cv. V1; Carbon Source 1. Introduction Mulberry (Morus spp.) is a woody perennial tree of im- portance to the sericulture industry as mulberry leaf is the sole food for the silkworm (Bombyx mori L.) larvae. Ow- ing to its long juvenile period and heterozygosity [1], im- provement of specific characters through conventional breeding is cumbersome and time consuming. Therefore targeted manipulation of elite genotypes through incor- poration of specific genes encoding desired traits using modern biotechnological methods offers a new opportu- nity for crop improvement. An efficient in vitro regenera- tion procedure is pre-requisite for transgenic approach in any crops. Information on development and standardiza- tion of in vitro regeneration protocols in promising mul- berry genotypes is limited, although there are reports in a few genotypes. Studies have been made in mulberry to examine the impact of various growth regulators on in vitro organogenesis and plant regeneration by using dif- ferent explants viz. leaf, internodal segment, hypocotyls and cotyledons [2-12]. However, the shoot differentiation from callus is confined only to a few genotypes and re- peatability of protocols developed was not assured due to the recalcitrant nature of the plant. In this study, we made an attempt to develop and standardize in vitro regenera- tion protocol in a widely cultivated mulberry variety, V1 using leaf explants. The variety is highly popular due to its economic characters under irrigated conditions and the foliage is suitable for both young and late age bivol- tine silkworm rearing. The major emphasis was given to investigate the effect of Thidiazuron (TDZ), a substituted phenyl urea and different carbon sources in inducing ad- ventitious buds and efficient regeneration in V1. * Corresponding author. Copyright © 2013 SciRes. AJPS