沟叶结缕草组织培养表面灭菌方法的筛选

目的]为建立沟叶结缕草再生体系奠定基础。方法]以沟叶结缕草带芽茎段为外植体,通过正交试验研究0.1%升汞、5%次氯酸钠、75%酒精和预处理方法等因素对外植体成活率的影响,筛选最佳表面灭菌方法。结果]各因素对外植体成活率的影响顺序为0.1%升汞>5%次氯酸钠>75%酒精>预处理方法,而0.1%升汞和5%次氯酸钠对其影响分别达到极显著和显著水平。沟叶结缕草组织培养的最佳表面灭菌方法为:先用5 g洗洁精加水冲洗2次,再使用5%次氯酸钠(滴入3滴100%吐温80),振荡灭菌10 min;最后用0.1%升汞(滴入3滴100%吐温80),振荡灭菌10 min。结论]可用杀菌剂、抗生素等对外植体进行预处理或将抗生素加入培养基中,起到杀菌和抑菌的作用。

Screening of Surface Sterilization Methods in the Tissue Culture of Zoysia matrella

Objective] The research aimed to lay the foundation for establishing the regeneration system of Zoysia matrelia.Method] With stem with buds of Zoysia matrella as explants,the effects of such factors as 0.1%mercuric chloride,5%sodium hypochlorite,75%alcohol and pretreatment methods on the survival rate of explants were studied though the orthogonal test.And the optimum surface sterilization methods was screened out.Result] The effects of each factor on the survival rate of explants orderly were 0.1%mercuric chloride >5%sodium hypochlorite >75%alcohol > pretreatment methods and the effects of 0.1%mercuric chloride and 5%sodium hypochlorite reached at extremely significant level and at a significant level.The optimum sterilization method in the tissue culture of Zoysia matrella was as follows: flushing with 5 g detergent and water for 2 times, and sterilizing with 5%sodium hypochlorite(dropping 3 drops of 100%Tween 80) for 10 min under vibratory conditions and finally sterilizing with 0.1%mercuric chloride(dropping 3 drops of 100%Tween 80) for 10 min under vibratory conditions.Conclusion] It could play the role of sterilization and bacteriostasis to pretreat explants with bactericide,antibiotic and so on or adding antibiotic to the medium.