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林业科学 ›› 2010, Vol. 46 ›› Issue (10): 95-102.doi: 10.11707/j.1001-7488.20101016

• 论文 • 上一篇    下一篇

根癌农杆菌介导的淡紫拟青霉遗传转化体系的建立

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   

  1. 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
  • 收稿日期:2010-05-04 修回日期:2010-07-23 出版日期:2010-10-25 发布日期:2010-10-25
  • 通讯作者: 汪来发

Establishment of Agrobacterium tumefaciens-Mediated Transformation System of Paecilomyces lilacinus

Wang Xizhuo1;Piao Chungen1;Li Hong2;Wang Laifa1;Guo Minwei1;Li Yong1;Lui Xiaoli3   

  1. 1.Key Laboratory of Forest Protection;State Forestry AdministrationResearch Institute of Forest Ecology Environment and Protection CAF Beijing 100091;2.Research Institute of Forestry CAF Beijing 100091;3.School of Forestry and Landscape Architecture;Anhui Agricultural University Hefei 230036
  • Received:2010-05-04 Revised:2010-07-23 Online:2010-10-25 Published:2010-10-25

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: 1 000~2 400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。

关键词: 根癌农杆菌, 淡紫拟青霉, G-418, 遗传转化

Abstract:

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 106 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.

Key words: Agrobacterium tumefaciens, Paecilomyces lilacinus, G-418, transformation