本文關(guān)鍵詞： 寡雄腐霉 生防真菌 農(nóng)桿菌 遺傳轉(zhuǎn)化 egfp基因　出處：《微生物學(xué)報》2017年10期 　論文類型：期刊論文

【摘要】：【目的】寡雄腐霉(Pythium oligandrum Drechsler)是一種對動、植物和環(huán)境無害,兼具殺菌和增產(chǎn)效果的生防真菌。通過研究建立農(nóng)桿菌介導(dǎo)的寡雄腐霉遺傳轉(zhuǎn)化體系。【方法】選用EHA105、AGL-1、LBA4404三種農(nóng)桿菌菌株對寡雄腐霉進(jìn)行遺傳轉(zhuǎn)化研究,通過對影響遺傳轉(zhuǎn)化效果的條件參數(shù)試驗(yàn)優(yōu)化,確立適宜寡雄腐霉遺傳轉(zhuǎn)化的農(nóng)桿菌菌株及轉(zhuǎn)化條件,建立農(nóng)桿菌介導(dǎo)的寡雄腐霉遺傳轉(zhuǎn)化體系。【結(jié)果】經(jīng)研究發(fā)現(xiàn),所選3種農(nóng)桿菌菌株中EHA105菌株對寡雄腐霉的遺傳轉(zhuǎn)化效果最好,其次是AGL-1菌株,LBA4404菌株轉(zhuǎn)化效果不好。EHA105菌株經(jīng)IM(含300μmol/L AS)誘導(dǎo)培養(yǎng)至OD_(600)=0.6時,與濃度為10~6 10~7個/m L的寡雄腐霉孢子懸浮液以1 10:1的比例混合,在25 26°C以液體振蕩的方式避光共培養(yǎng)72 h(pH 5.0,含300μmol/L AS),寡雄腐霉菌體液體振蕩恢復(fù)培養(yǎng)24 h,涂布抗性選擇平板篩選寡雄腐霉轉(zhuǎn)化子,即可得到寡雄腐霉基因工程菌株,其轉(zhuǎn)化率可達(dá)到130個轉(zhuǎn)化子/106個孢子�！窘Y(jié)論】本研究首次構(gòu)建了農(nóng)桿菌介導(dǎo)的寡雄腐霉遺傳轉(zhuǎn)化體系,研究結(jié)果可為寡雄腐霉的生防機(jī)制及分子育種研究提供技術(shù)支撐。
[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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