绿僵菌IMI330189菌株突变体库的建立及毒力相关突变株的筛选

为了研究绿僵菌毒力相关的功能基因，采用农杆菌介导的转化方法将携带苯菌灵抗性标记基因的T—DNA导入绿僵菌IMI330189中，获得了绿僵菌突变体库，采用人工饲料诱饵法测定了突变株对3龄东亚飞蝗Locustamigratoria的毒力。研究发现，突变株对东亚飞蝗的毒力都有不同程度的改变，在诱饵剂浓度为10^8孢子．g-1下，野生型菌株IMI330189对东亚飞蝗的致死中时（LT50）为4-3d；突变株189-1的毒力略有提高，其LT50值为3．5d；其余突变株的毒力均降低，且突变株189—4、189—5、189—6、189—10、189-11、189—13和189—15几乎丧失了对东亚飞蝗的毒性，校正死亡率小于50％。本研究为进一步分离毒力相关基因、探索绿僵菌的致病机制奠定了基础。

Construction of Metarhizium anisopliae IMI330189 Mutant Library and Screening of Strains Related to Virulence

In order to explore the functional genes of Metarhizium anisopliae related to virulence, a T-DNA containing benomyl resistance gene benA3 was transferred into M. anisopliae strain IMI330189 to construct mutant library by using Agrobacterium-mediated transformation method. The toxicity of some mutants to the third instar larvae ofLocusta migratoria was tested at a concentration of 108 spores.g-l with artificial bait method. The toxicity of mutant 189-1 was slightly shorter with the LTs0 3.5 d compared to the wild strain IMI330189 with LT50 4.3 d. The mutants 189-4, 189-5, 189-6, 189-10, 189-11, 189-13 and 189-15 almost lost their toxicity to locust, the corrected mortality rates were less than 50%. By constructing mutant library of M. anisopliae strains IMI330189, we obtained some mutants with virulence lack. The study provided a tool to isolate virulent genes and explore the pathogenic mechanism of M. anisopliae.