锯叶班克木Banksia serrata外植体的选择及消毒方法的研究

Study on selection and sterilization of explants of Banksia serrata in tissue culture

锯叶班克木为山龙眼科班克木属的木本花卉,全株密被绒毛是影响组培启动培养的主要原因之一。为成功建立锯叶班克木的组培体系,系统地研究了外植体类型、取材方法、灭菌方法等对启动培养外植体消毒效果的影响。结果表明,10-12月份采取的枝条最适合做外植体,污染率在30%以下,外植体存活率最高可达63.3%;半木质化枝条的中上部为最佳的外植体取材部位,顶芽虽然污染率低但存活率也低,下部茎段污染率较高,均不适宜作为外植体材料;外植体灭菌的最佳方法为:用1 000倍的多菌灵溶液清洗外植体然后用4%NaClO+0.01%吐温80处理12 min,启动培养污染率可降低至26.7%。酒精和HgCl2均可在短时间内杀死外植体,不适合用于外植体的灭菌。

Banksia serrata is one of the perennial woody species belonging to Proteaceae family.Its surface is densely tomentose,resulting in extremely high pollution rate of explants in tissue culture.The effects of explants types,seasons,bud positions,and sterilization method on explants survival were studied.The results show that the optimal months to take explants were from October to December,with the pollution rate below 30% and the survival rate of 63.3%.The optimal explants were cut from the upper part of semilignified softwood stems.The best sterilization method was that the explants were washed with 1 000 times dilution of carbendazim for three times,and then dipped in 4% NaClO ＋ 0.01% Tween 80 for 12 min,and the explants pollution rate was reduced to 26.7%.However,both alcohol and HgCl2solutions killed the explants in a short time,and was not suitable for explants disinfection.