农杆菌介导的携带egfp基因载体转化寡雄腐霉

发布时间：2018-03-01 01:31

  本文关键词： 寡雄腐霉 生防真菌 农杆菌 遗传转化 egfp基因　出处：《微生物学报》2017年10期 　论文类型：期刊论文

【摘要】：【目的】寡雄腐霉(Pythium oligandrum Drechsler)是一种对动、植物和环境无害,兼具杀菌和增产效果的生防真菌。通过研究建立农杆菌介导的寡雄腐霉遗传转化体系。【方法】选用EHA105、AGL-1、LBA4404三种农杆菌菌株对寡雄腐霉进行遗传转化研究,通过对影响遗传转化效果的条件参数试验优化,确立适宜寡雄腐霉遗传转化的农杆菌菌株及转化条件,建立农杆菌介导的寡雄腐霉遗传转化体系。【结果】经研究发现,所选3种农杆菌菌株中EHA105菌株对寡雄腐霉的遗传转化效果最好,其次是AGL-1菌株,LBA4404菌株转化效果不好。EHA105菌株经IM(含300μmol/L AS)诱导培养至OD_(600)=0.6时,与浓度为10~6 10~7个/m L的寡雄腐霉孢子悬浮液以1 10:1的比例混合,在25 26°C以液体振荡的方式避光共培养72 h(pH 5.0,含300μmol/L AS),寡雄腐霉菌体液体振荡恢复培养24 h,涂布抗性选择平板筛选寡雄腐霉转化子,即可得到寡雄腐霉基因工程菌株,其转化率可达到130个转化子/106个孢子。【结论】本研究首次构建了农杆菌介导的寡雄腐霉遗传转化体系,研究结果可为寡雄腐霉的生防机制及分子育种研究提供技术支撑。
[Abstract]:[objective] Pythium oligandrum Drechsler is a species that is harmless to animals, plants and environment. The genetic transformation system of Pythium oligaricus mediated by Agrobacterium tumefaciens was established. [methods] three Agrobacterium strains, EHA105AGL-1 and LBA4404, were selected to study the genetic transformation of Pythium oligodui. Through the optimization of the condition parameters affecting the effect of genetic transformation, the Agrobacterium tumefaciens strain suitable for genetic transformation of Pythium oligaricus and its transformation conditions were established, and the Agrobacterium tumefaciens mediated transformation system of Pythium oligoandroides was established. [results]. Among the 3 strains of Agrobacterium tumefaciens, EHA105 strain had the best effect on genetic transformation of Pythium oligodon, followed by AGL-1 strain LBA4404. EHA105 strain was induced to OD_(600)=0.6 by IMI (containing 300 渭 mol/L ASS). Mixed with the spore suspensions of Pythium oligoandrae with a concentration of 10 ~ 6 ~ 10 ~ 7 / mL at a ratio of 1 / 10: 1, At 25 鈩，

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