根癌农杆菌介导的淡紫拟青霉遗传转化体系的建立  

Establishment of Agrobacterium tumefaciens-Mediated Transformation System of Paecilomyces lilacinus

作　　者：王曦茁[1] 朴春根[1] 李虹[2] 汪来发[1] 郭民伟[1] 李永[1] 刘晓莉[3] 

机构地区：[1]中国林业科学研究院森林生态环境与保护研究所国家林业局森林保护学重点实验室,北京100091 [2]中国林业科学研究院林业研究所,北京100091 [3]安徽农业大学林学与园林学院,合肥230036

出　　处：《林业科学》2010年第10期95-102,共8页Scientia Silvae Sinicae

基　　金：国家自然科学基金(30771731);农业部科技成果转化资金项目(2010GB24320615)

摘　　要：基于根癌农杆菌介导的遗传转化方法,选择植物线虫生防真菌淡紫拟青霉的分生孢子为受体,建立以G418为筛选标记遗传转化体系。通过优化遗传转化条件,达到较高的转化效率:1000～2400个转化子·10-6分生孢子。对转化子进行PCR验证,表明T-DNA已整合到淡紫拟青霉的基因组中,表型测定发现转化子的遗传表现稳定。农杆菌介导淡紫拟青霉突变体库的建立,为进一步筛选对植物线虫致病性高的突变体、了解淡紫拟青霉的侵染过程和侵染机制提供基础,对培育更优良的植物线虫生防菌株,开发高效生防制剂等具有重要的意义。In this study,the research is conidia of Paecilomyces lilacinus,a pathogenic fungus of plant nematodes,was selected as the receptor. An efficient genetic transformation system of P. lilacinus was established by harboring G-418 resistance cassette as a selective marker to construct vector pBI-G3C-aphI based on Agrobacterium tumefaciens-mediated transformation. Using this system,P. lilacinus was successfully transformed with an efficiency of 1 000-2 400 transformants per 10^6 spores. It indicated that T-DNA had been integrated into the genome of P. lilacinus by PCR analysis on the transformants,and the genetic character of transformants remained stable. The establishment of T-DNA insertional mutants library would provides a basis for further selecting pathogenic mutants,and understanding pathogenisis of P. lilacinus,which is of great significance to foster a better anti-nematode strain and develop effective biocontrol agents.

关 键 词：根癌农杆菌 淡紫拟青霉 G-418 遗传转化 

分 类 号：S763.1[农业科学—森林保护学]