崇左金花茶的外植体灭菌研究

Study on Explants Sterilization of Camellia chungtsoensis S.Y.Liang et L.D.Huang

以崇左金花茶的幼嫩叶片、幼嫩茎段、半木质化茎段、芽、花苞和种子为外植体,采用75%酒精、0.1%HgCl2、0.2%HgCl2和NaClO 4种灭菌剂进行消毒灭菌处理,研究了崇左金花茶组织培养中外植体的最佳灭菌方案。结果表明:幼嫩叶片和种子的消毒相对容易,幼嫩叶片采用75%酒精30s+0.1%HgCl25min+0.1%HgCl25min的灭菌方法好;种子采用75%酒精40s+0.1%HgCl220～30min+0.2%HgCl220min的灭菌方法好。茎段的消毒相对较难,采用分段灭菌的方法效果较好,幼嫩茎段采用75%酒精25s+0.1%HgCl210min+0.2%HgCl25min的灭菌方法好;半木质化茎段采用75%酒精35s+0.1%HgCl212min+0.2%HgCl26min的灭菌方法好。芽和花苞的消毒最难,采用NaClO原液+HgCl2分段灭菌,并逐次剥去外层苞片进行灭菌的方法也难以达到预期效果。

Taking the young leaves, the tender stem segments, the half lignified stem segments, the leaf buds, the buds and the seeds of Camellia chungtsoensis S. Y. Liang et L. D. Huang as explants,with 75% alcohol,0. 1% HgCl2 ,0. 2% HgCl2 and NaC10 4 kinds of sterilization agent for sterilization, explants in tissue culture of Camellia chungtsoensis S. Y. Liang et L. D. Huang optimal sterilization program were studied. The results showed that it was easy to the young leaves and the seeds to disinfect. The better disinfection condition for the young leaves was 75 % alcohol sterilized for 30 s, 0. 1% HgCl2 of mercury disinfected for 5 minutes and 0. 1% HgCl2 of mercury disinfected for 5 minutes again. The better disinfection condition for the seeds was 75 % alcohol sterilized for 40 s,0. 1% HgCl2 of mercury disinfected for 20 to 30 minutes and then 0. 2% HgCl2 of mercury disinfected for 20 minutes. It wasn＇t easy to the stems to disinfect,the better way was time period of disinfection. The better disinfection condition for the young stems was 75 % alcohol sterilized for 25 s,0. 1% HgCl2 of mercury disinfected for 10 minutes and then 0. 2% HgCle of mercury disinfected for 5 minutes. The better disinfection condition for the half lignified stems was 75% alcohol sterilized for 35 s, 0. 1% HgC12 of mercury disinfected for 12 minutes and then 0. 2% HgC12 of mercury disinfected for 6 minutes. And difficult to the buds. It was difficult to achieve the desired effect of time period of disinfection with NaC10＋ HgC12 and successive strip the outer bracts sterilization method.