Restriction enzyme-mediated integration(REMI) of DNA has been recently received attention as a new technique for the generation of mutants by transformation in fungi. Trichoderma atroviride strain T23 was transformed ...Restriction enzyme-mediated integration(REMI) of DNA has been recently received attention as a new technique for the generation of mutants by transformation in fungi. Trichoderma atroviride strain T23 was transformed with linearized plasmid pV2, conferring resistance to hygromycin B, in the presence of restriction enzyme used to linearize the plasmid. A total of 172 regeneration transformants were detected by successive inoculation for seven times subcultivation on fresh PDA plate containing hygromycin B. The plasmid was integrated stably into the chromosome DNA, which was confirmed by PCR and southern analysis. The difference between 172 transformants and the parent strain was confirmed in colonial color, sporulation and growth rate. The results showed that the significant difference appeared in above mentioned characters between transformants and parent strain is sporulation capability. Transformants TC6, TD5, TE7, TF1 and TK1 produced higher amounts of conidia than the parent strain T23. In addition, transformants TK1and TC6 showed stronger inhibition to the growth rate of the cucumber wilt pathogen (Fusarium oxyporum) in vitro.展开更多
利用限制性内切酶介导整合(REM I)技术,通过插入线性化质粒DNA获得了生物防治番茄灰霉病(B otry tis cinerea)效果优于初发菌T richod erm a v irid e T 21菌株(初发菌)的3个变异株T trm 31、T trm 34和T trm 55.变异株本身产生的和诱...利用限制性内切酶介导整合(REM I)技术,通过插入线性化质粒DNA获得了生物防治番茄灰霉病(B otry tis cinerea)效果优于初发菌T richod erm a v irid e T 21菌株(初发菌)的3个变异株T trm 31、T trm 34和T trm 55.变异株本身产生的和诱导番茄植株产生的几丁质酶和-β1,3-葡聚糖酶的活性均比初发菌高,因此,通过REM I技术可以获得新的有益变异株,在一定程度上提高了生物菌株防治番茄灰霉病的水平,尤其是提高诱导番茄抗病水平.对侵染花器和叶片的灰霉病防效分别比原生物防治木霉菌株提高了16.9%和8%,说明REM I技术可以用于改良生防木霉菌株的功能,提高生物防治效果.展开更多
基金Item supported by the Tenth Five-yearProgram of science and technology for creation of biocontrol a-gent against corn disease(2004BA509B0405)
文摘Restriction enzyme-mediated integration(REMI) of DNA has been recently received attention as a new technique for the generation of mutants by transformation in fungi. Trichoderma atroviride strain T23 was transformed with linearized plasmid pV2, conferring resistance to hygromycin B, in the presence of restriction enzyme used to linearize the plasmid. A total of 172 regeneration transformants were detected by successive inoculation for seven times subcultivation on fresh PDA plate containing hygromycin B. The plasmid was integrated stably into the chromosome DNA, which was confirmed by PCR and southern analysis. The difference between 172 transformants and the parent strain was confirmed in colonial color, sporulation and growth rate. The results showed that the significant difference appeared in above mentioned characters between transformants and parent strain is sporulation capability. Transformants TC6, TD5, TE7, TF1 and TK1 produced higher amounts of conidia than the parent strain T23. In addition, transformants TK1and TC6 showed stronger inhibition to the growth rate of the cucumber wilt pathogen (Fusarium oxyporum) in vitro.
文摘利用限制性内切酶介导整合(REM I)技术,通过插入线性化质粒DNA获得了生物防治番茄灰霉病(B otry tis cinerea)效果优于初发菌T richod erm a v irid e T 21菌株(初发菌)的3个变异株T trm 31、T trm 34和T trm 55.变异株本身产生的和诱导番茄植株产生的几丁质酶和-β1,3-葡聚糖酶的活性均比初发菌高,因此,通过REM I技术可以获得新的有益变异株,在一定程度上提高了生物菌株防治番茄灰霉病的水平,尤其是提高诱导番茄抗病水平.对侵染花器和叶片的灰霉病防效分别比原生物防治木霉菌株提高了16.9%和8%,说明REM I技术可以用于改良生防木霉菌株的功能,提高生物防治效果.
