播娘蒿原生质体培养研究

Study on Protoplast Culture of Descurainia sophia

以播娘蒿无菌苗的子叶和下胚轴分离原生质体 ,每克鲜重产量最高达 ( 1～ 1 5 )× 10 6 个 .对原生质体进行液体浅层静置培养 .结果表明 :最佳酶解液酶组成为 4 %纤维素酶 + 2 %果胶酶 + 0 5 %离析酶 ;最佳培养密度为2 0× 10 4个 /mL ;最佳渗透压稳定剂为 10 %蔗糖 + 2 %葡萄糖 .观察表明 :播娘蒿原生质体 1～ 7d内出现第一次分裂 ,分裂频率约 2 0 %,培养 18d时具几十个细胞的细胞团 ,以后逐渐形成肉眼可见的小愈伤组织 .

The highest yield of purified protoplasts isolated from cotyledon and hypocotyl of sterile seeding of D.sophia could reach about 1～1.5×10 6/gFW.When the protoplasts were cultured in liquid medium,the research results showed that the best combination of the enzyme solution was 4 % Cellulase Onozuka R-10+2 % Pectinase+0.5 % Macerozyme R-10;the best dentsity of protoplast culture was 20×10 4/ml;and the best osmotic stabilizer was 10 % sucrose+2 % glucose.The first cell divison occurred within 1～7 days and the highest divison rate was about 20 %.Eighteen days later,a great unmber of cell colonies and microcalli could be observed.