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[目的]研究N+离子束注入对生防菌枯草芽孢杆菌存活率和突变率的影响。[方法]对离子束注入前样品处理方法进行优化,包括枯草芽孢杆菌液体培养时间、稀释浓度、稀释溶剂和菌膜干燥时间;采用N+离子束注入进行诱变处理,并通过平板对峙法和牛津杯法筛选菌株。[结果]N+离子束注入样品前处理最佳条件为:枯草芽孢杆菌液体培养20~24h,利用无菌水将菌液稀释到106个/ml,菌膜干燥时间为0~60min;离子束注入诱变的最佳条件为:注入能量30kev,剂量为2.0×1014~4.0×1014ions/cm2,枯草芽孢杆菌存活率为8.43%~26.71%,突变率为3.50%~5.43%。[结论]该研究结果为枯草芽孢杆菌离子束注入诱变育种方法的研究提供了参考依据,同时也为其他生防菌的辐射诱变育种奠定了基础。
[Objective] The research aimed to study the effects of N + ion beam implantation on the viability and mutation rate of Bacillus subtilis. [Method] The sample preparation method before ion beam injection was optimized, including liquid culture time, dilution concentration, dilution solvent and membrane drying time of Bacillus subtilis. The seeds were treated with N + ion beam and mutagenized by plate confrontation method and Oxford Cup method screening strains. [Result] The optimum conditions for pretreatment of N + ion beam were as follows: the Bacillus subtilis liquid was cultured for 20-24 h, the bacterial liquid was diluted to 106 / ml with sterile water, and the drying time of the membrane was 0-60min; The optimal conditions for the mutagenesis were as follows: 30 kev implantation dose, 2.0 × 1014 ~ 4.0 × 1014ions / cm2 dose, Bacillus subtilis survival rate of 8.43% ~ 26.71% and mutation rate of 3.50% ~ 5.43%. [Conclusion] The results of this study provide a reference for the study of mutagenesis breeding by ion beam implantation of Bacillus subtilis, and laid the foundation for radiation induced mutation breeding of other biocontrol agents.