蛇足石杉外植体表面消毒及内生菌消除方法

Sterilization of Explants and Elimination of Endophytes in Huperzia Serrata

为了获得蛇足石杉的无菌材料,使用双氧水、酒精、升汞等消毒剂对外植体进行表面消毒,使用混合抗菌溶液及不同温度的热水浴处理外植体以消除内生菌.结果表明:茎段经预处理后,用47℃热水浴处理1 h,再经70%酒精处理30 s,0.1%升汞处理7 min后,能达到较好的表面消毒及内生菌消除的效果;孢子囊和茎尖经预处理后,再经过70%酒精处理30 s,0.1%升汞处理4 min即可达到较好的表面消毒效果;消毒剂结合温度处理能降低内生菌的污染率;孢子囊和茎尖几乎不含内生菌,是可用于组织培养的较好的外植体.

In order to obtain sterile materails from Huperzia serrata, H2O2 ,C2H5OH, and HgCl2 were used to disinfect the explants, and the solutions of antibiotic compound together with hot water treatments were used to eliminate the endophytes. The results were as folloWs： the optimum method of the sterilization of surface and elimination of endophytes for stem was obtained by soaking the explants in hot water （47 ℃） for 1 h,followed by 70% C2H5OH for 30 s and then by 0.1 % HgCl2 for 7 min. The optimum method of the sterilization of surface for sporangium and stem apex was obtained by soaking the explants in 70 % C2H5OH for 30 s, followed by 0.1% HgCl2 for 4 min. Disinfectants together with hot water treatment can obviously reduce the pollution rate of explants. Sporangium and stem apex,which barely contain endophytes, sre excellent explants for plant tissue culture.