Optimization of protocols for callus induction, regeneration and acclimatization of Sugarcane (Saccharum officinarum L.) cultivar CO-86032

Abstract

An efficient protocol for induction of callus and regeneration of a high yielding sugar cane var CO-86032 has been developed and reported here. Callus induction from leaf sheath explants derived from 2-3-month-old plants was achieved on Murashige and Skoog's(MS) medium supplemented with different auxins viz, 2,4-D, 2,4,5-T, NAA, dicamba and picloram (1-3 mgl-1). Among different auxins, 2,4-D at 1 mgl-1 supplemented with 2% sucrose + 300mgl-1 PVP was found favorable in inducing callus and preventing browning. Addition of coconut milk and Kn further enhanced the growth of callus maximum being on MS medium supplemented with 0.5mg1-1Kn (1994±0.39mg). Calli were further evaluated for regeneration. MS medium supplemented with 2mg/l Kn+1mg/l BAP was found suitable where 100% calli regenerated with maximum number of multiple shoots per callus mass(168±0.54). Highest number of root emergence (38±0.21) and maximum root length (6.8±0.64cm) was achieved on MS medium supplemented with 5mgl-1 NAA. The in vitro grown plants were transferred to polycups containing a mixture of sterilized sand and black soil (1:1) for hardening. The hardened plants were transferred to green-house conditions where they survived with 90% frequency.