匍匐翦股颖和狗牙根原生质体融合研究

以匍匐翦股颖克罗米和狗牙根新农1号为材料,通过愈伤诱导、继代和悬浮培养,进而细胞融合来探讨草坪草品种改良的新方法.结果表明:(1)匍匐翦股颖和狗牙根原生质体融合时酶解液的最佳组合为:1.5%纤维素酶、0.1%果胶酶、0.7M甘露醇,10h酶解时间,且酶解前应用高渗溶液预处理效果更佳;(2)细胞融合时,原生质体密度2×106个/mL,融合剂由30%的PEG4000、15mmol/L CaCl2·2H2O、0.5mmol/L KH2PO4·H2O、0.5mol/L甘露醇组成,并调pH至7.5效果较好;(3)匍匐翦股颖和狗牙根细胞PEG融合率约为5%,以30min融合时间效果较好,产生的多核融合体少,杂种细胞多呈条形,具有较强的活力.

Study on Protoplast Fusion of Creeping Bentgrass and Bermuda Grass

After several generations of successive transferation and suspension culture, the best sus- pended cell materials of Creeping Bentgrass and Bermuda grass were selected. Their cell wall were re- moved and carried on the protoplast fusion. The objective of this research was to find a new method to im- prove turf quality. The results indicated that. (1) The optimal enzymolysis solution involved 1.5% cellu- lases, 0.1% pectolase, and 0.7M mannitol; and 10 hours was the best enzymolysis time. In addition, pre- treatment of the protoplast in sugar solution before the enzymolysis had better effect. (2) The fusion agents included 30% PEG4000,15mmol/L CaCl2·2H2O,0.5mmol/L KH2PO4·H2O and 0.5mol/L mannitol, and pH 7.5 . Before fusion, adjusted the protoplast volume density to 2×105/mL. (3) The cell fu- sion rates of the two turf materials was 50%, 30min of fusion time would be better to reduce coenocytia, the hybrid cells seemed to be monstrous but with high vitality.