蛇足石杉茎尖灭菌方法与组织培养的研究  

Studies on Tissue Culture and Sterilization Method of Huperzia serrata

作　　者：杨雪飞[1] 罗建平[1] 王瑛[1] 

机构地区：[1]合肥工业大学生物与食品工程学院,安徽合肥230009

出　　处：《安徽农业科学》2008年第12期4947-4948,5025,共3页Journal of Anhui Agricultural Sciences

基　　金：安徽省自然科学基金项目(00041508)

摘　　要：[目的]研究蛇足石杉组织培养的灭菌方法和培养条件。[方法]以野生孢子体茎尖为外植体,先用不同灭菌液单独或配合使用对外植体进行表面灭菌,将表面无菌外植体转到添加多种抗生素和孔雀石绿的培养基上杀灭内生菌,然后将外植体分别接种到不同的培养基上,筛选茎尖培养的最佳灭菌方法和培养条件。[结果]最佳表面灭菌方法为70%乙醇灭菌40 s,0.1%升汞溶液灭菌8 min,70%双氧水溶液灭菌10 min,培养2周,表面无菌率为63%。适宜的杀灭内生菌方法为表面无菌的外植体在附加0.5 mg/L孔雀石绿和100 mg/L抗生素AAS的培养基上培养4~6周,无菌率达52%。适宜的培养基为1/4MS矿质元素＋1/2MS有机元素＋2%蔗糖＋4.5 g/L琼脂,pH5.8。在此培养基上茎尖外植体可生长出新枝,并可再生根。[结论]建立了蛇足石杉茎尖组织培养的灭菌方法且筛选出适合其生长的适宜培养基,为蛇足石杉组织培养的深入研究提供实验基础。[Objective] To study the culture condition and sterilization method of Huperzia serrata.[Method]Shoot tips of 1.0～1.5 cm of wild sporophytes were selected as explants,which underwent surface sterilization singly or combined with various disinfectors and internal sterilization with antibiotics and Malachite Green to eliminate the epiphytic and endophytic bacteria and fungi and were next placed on different modifications of MS medium to culture.[Results] Optimum sterilization method was 70% C2H5OH for 40 s followed by 0.1% HgCl2 for 8 min and subsequent 70% H2O2 for 10 min,and aseptic ratio of 63% was reached.The sterilized explants were cultured on medium with 0.5 mg/L Malachite Green,100 mg/L antibiotic AAS for 4 weeks and 52% of aseptic explants were achieved.The optimum medium for explant growth and rooting contained mineral elements of MS at quarter strength,organic elements of MS at half strength,2% sucrose and 4.5 g/L Agar.[Conclusion] Using shoot-tip as explants,tissue culture and sterilization meth od of Huperzia serrata were established.

关 键 词：蛇足石杉 孢子体 组织培养 灭菌 

分 类 号：S722.37[农业科学—林木遗传育种]