美国山核桃组培中材料灭菌的研究

Sterilization of Carya illinoensis Explants in Tissue Culture

采用不同的灭菌措施,包括不同的灭菌剂、浓度和处理顺序,对美国山核桃带芽茎段进行灭菌处理,研究其污染率和污染速率的变化,寻找建立无菌培养系最适宜的方法。结果表明,外植体材料最适宜的灭菌处理方法为:先将外植体在70%酒精中浸30s,再将其用0.2%升汞液(加数滴吐温80)浸15min,无菌水冲洗10次,最后接种。培养基中添加适量活性炭,且前期暗培养是提高材料无菌率的有利保障。另外,培养基中加入青霉素、链霉素各100mg·L-1可明显提高材料的无菌率。

Carya illinoensis K.Koch stems with leading bud were sterilized with different sterilizing measures, including different sterilizing pharmaceutical in different densities and different orders.The most suitable method for asepsis cultivation was set up by analysis the pollution rate and speed.Results were as follows:the most suitable sterilization method of explant was soaking them in 70% alcohol for 30 s,putting them in 0.2% HgCl(adding several drops of Tween 80) for 15 mins and then washed 10 times by asepsis water.Adding right active charcoal in medium and no light during the early stage was benefit for the rate of asepsis;Putting penicillin, streptomycin (100 mg·L^(-1)) into medium could improve asepsis rate obviously.