摘要
利用农杆菌T-DNA介导的遗传转化体系转化稻瘟病菌(Magnaporthe grisea)Y34菌株,平均每转化1.0×106个稻瘟病菌分生孢子可得到约300个抗潮霉素菌株。用该转化体系转化和筛选,获得抗潮霉素菌株6855个。PCR检测结果表明,所有表现抗潮霉素菌株均含抗潮霉素基因,说明抗潮霉素特性是T-DNA携带潮霉素基因插入Y34基因组的表型效应,即抗潮霉素菌株是T-DNA插入突变体。对56个突变体DNASouthern检测结果表明,有27个突变体是单拷贝插入,突变体T-DNA插入拷贝数平均为1.43。随机取1600个突变体进行致病力测定,结果发现23个突变体完全丧失致病能力。
According to the transformation system established by our laboratory, Magnaporthe grisea strain Y34 was mutated through T-DNA insertion mediated by Agrobacterium turnefaciens strain AGL-1. The transformation efficiency was 300 transformants per 1.0 ×10^6 conidia of the fungus on average, and 6 855 transformants were obtained. PCR amplification found that all tested transformants were resistant to hygromycin and contained hph gene, indicating that the transformants were T-DNA inserted mutants. DNA Southern blot analysis of 56 T-DNA inserted mutants indicated that the copy number of T-DNA insertion was 1.43, and that 27 mutants were singlecopy insertions. Pathogenicity test of randomly selected 1 600 transformants found that 23 mutants completely lost their virulence on a susceptible rice cultivar.
出处
《热带作物学报》
CSCD
2007年第1期80-84,共5页
Chinese Journal of Tropical Crops
基金
国家自然科学基金资助项目(30260046)
中国热带农业科学院科技基金
