甜菜原生质体培养及大块愈伤组织的产生

Sugarbeet protoplast culture and the Formation of big callus tissures

将发芽盒内培养 2～ 3周的甜菜子叶 ,在CPW 1 2M高渗溶液中预质壁分离 4h后用 2 %CellulaseOnozukK - 1 0 +0 .5%Macerozyme +0 .5%Driselase +CPW 9M酶液酶解 1 2h。原生质体在K8P +2 4-D2 0mg/L +6-BA 0 .5mg/L培养基中液体浅层培养。 6周内形成大量的多细胞团和微愈伤组织 ,进一步培养 ,形成直径为 0

Cotyledons were cut down from sugarbeet plant which were cultured in germination box between two and three weeks.To get protoplasts,cotyledons which were cut into pieces were soluted in CPW 12M media for 4 hours,in enzyme mixture for 12h,enzyme mixture consists of 2.0% cellulase Onozuka R-10+0.5% Macerozyme Onozukar-10+0 5% Driselase+CPW 9M.The isolutated protoplast were cultured in K8P+NAA 2.0mg/L+BA 0.5mg/L media.In six weeks,cell cluster an microscopical callus were formated.Big loose callus were obtained in following days.