Effect of Sorbitol Pretreatment On Yield and Viability of Protoplasts isolated From Etiolated Shoots of Three :Cultivars of Sorgi/Um Bicolor
This study aimed at improving Sorghum protoplasts isolation procedures and maximizing yield . using etlolated shoots of three cultlvars (MH61, CH9 and CVX) fer Sorghum blcolor (L) Moench. Hypocotyl sections were pretreated in sorbltol for "arying periods of time and protoplast Isolated In medium (pH 5.7) containing 3% cellulase (ONOZUKA RS.), 0.5% mecerozyme (R-10) and 0.1% pectolyase (Y-23) in protoplast wash solution. Pretreatment time of 3.5 hours In sorbitol was found to be optimal for protoplast yield and vlablllty. Optimal protoplast yields of 9.22 x 106, 8.10 x 105 and 6080 x 105 were obtained per gram fresh weight of hypocotyl tissue in cultivara MH51, CH9 and CVX respectively, at 2.6hour digestion time. This low enzyme digestion time decreased the. toxic effects of enzyme preparations and:. consequently enhanced viability up to 75%. Results show that etiolated seedlings can provide a rapict and high-yietding source of viable Sorghum protoplasts. Efforts should, therefore, be geared towards improving the plating efficiency and regeneration capacity of protoplasts .