蛇足石杉的组织培养初探

Tissue Culture of Huperzia Serrata

针对蛇足石杉资源分散,资源更新周期长,难以大规模开采的情况,初步探讨了蛇足石杉的组织培养技术.以蛇足石杉茎尖为外植体,分别在1/2MS+0.05μmol/L IBA+1.4μmol/L KT+2 mg/L谷氨酰胺、MS+0.05μmol/L IBA+1.4μmol/L KT+2 mg/L谷氨酰胺、1/2MS+2 mg/L谷氨酰胺/无激素、MS+2 mg/L谷氨酰胺/无激素等4种培养基中培养.结果显示:较为适宜的初代培养基为2号培养基,即MS+0.05μmol/L IBA+1.4μmol/L KT+2 mg/L谷氨酰胺.在组织培养工作中,污染问题比较严重,本实验中,采用了以下2种方法对外植体进行消毒:(1)70%酒精浸泡1 min→5%NaOCl浸泡1 min→7%H2O2浸泡10 min;(2)0.1%升汞液灭菌3’→无菌水冲洗5次.结果显示第2种方法比较理想.

In this study, tissue culture was explored to solve the propagate problem in the production with Huperzia serrata （Thunb）Trev. The stem and shoot-tips were used as explants to induce callus. These explants are cultured in four culture mediums： （1） 1/2MS＋0.05 μmol/L IBA＋1. 4μmol/L KT＋2 mg/L pirydoxine; （2） MS＋0.05 μmol/L IBA＋1.4 pμmol/L KT＋2 mg/L pirydoxine; （3） 1/2MS＋2 mg/L pi- rydoxine;（4） MS＋2 mg/L pirydoxine. The results show that the suitable element compositions to induce callus is：MS＋0.05 μmol/L IBA＋ 1.4μmol/L KT＋2 mg/L pirydoxine.During tissue culture, fungal pollution is very serious, so the surface disinfection are two variants of surface sterilization. In the first variant shoot-tips were soaked is important. There 70% ethyl alcohol （C2HsOH） for 1 min,5% sodium hypochlorite （NaOC1） for 1 min and then in 7% hydrogen peroxide （H202） for 10 min;the second variant were soaked in HgC1 for 3 min. The result shows that the most suitable procedure for surface sterilization of explants is the second one.