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[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。
[Objective] The research aimed to establish the foundation for the regeneration system of Zoysia matrella. [Method] With the stem of Buds of Zoysia matrella as explants, the effects of 0.1% mercuric chloride, 5% sodium hypochlorite, 75% alcohol and pretreatment methods on the survival rate of explants were studied by orthogonal test. The best method of surface sterilization. [Result] The order of influence of various factors on the survival rate of explants was 0.1% mercuric chloride> 5% sodium hypochlorite> 75% alcohol> pretreatment method, while the effects of 0.1% mercuric chloride and 5% sodium hypochlorite reached extremely significant and significant . The best method of surface sterilization for tissue culture of Zoysia matrella was as follows: rinsing with 5 g detergent and water twice, then using 5% sodium hypochlorite (dropping 3 drops of 100% Tween 80) and shaking and sterilizing for 10 min ; Finally, 0.1% mercuric chloride (3 drops of 100% Tween 80) was added dropwise and shaken for 10 min. [Conclusion] The explants could be pretreated with antibacterial agents and antibiotics or added antibiotics to the medium, which could kill bacteria and inhibit bacteria.