Optimization of surface sterilization and manipulation of in vitro conditions for reduced browning in pomegranate (Punica granatum L.) variety Bhagava

Abstract

Success in commercially viable protocol for mass multiplication of plants using tissue culture begins with effective sterilization and reducing exudation from perennial fruit crops which makes the in vitro techniques more difficult. In the present study, nodal explants of pomegranate cv. Bhagva were established using different durations and concentrations of anti-microbial agents followed by treatments with anti-oxidants (citric and ascorbic acid), with and without illumination (light and dark) and presence or absence of activated charcoal in MS media for reducing browning. Among different treatments tested for sterilization, 300 mgl-1 bavistine (50% carbendazim) for eighteen mintues, 200 mgl-1 each of streptocycline and cefotaxime for eighteen and eight minutes, respectively were found to be most the effective treatments to obtain cent per cent axenic culture development. Incorporation of activated charcoal (0.25%) significantly reduced the browning and improved the establishment of cultures as compared to other anti-browning agents.