Cryopreservation of shoot-tips from different sugarcane varieties using D cryo-plate technique

Abstract

Cryopreservation is an effective approach to conserve sugarcane germplasm for long term. This study was conducted to develop D cryo-plate technique to conserve sugarcane’s shoot tips. This new and effective cryopreservation technique will serve the purpose to conserve sugarcane germplasm efficiently on the large scale at genebanks. Sugarcane variety Ni-1 were used to optimize the protocol and after optimization 11 other varieties were tested with optimized protocol. It was observed that shoot tips of Ni-1 with a length of 1.5 to 2.0 mm, precultured on semi-solid 1/2 MS medium for 1 day and semi-solid MS medium with 0.3 M sucrose for 1 day, treated with loading solution containing 2.0 M glycerol + 1.2 M sucrose for 30 min and air dehydrated in an air laminar flow’s air current for 45 min, removed from alginate gel after cryopreservation and kept in dark for seven days produced maximum regrowth (97.7%). Range of 20.0% to 100% with an average of 52.1% was observed among sugarcane varieties for regrowth after cryopreservation with optimized D cryoplate protocol.