稻瘟病菌限制酶介导整合(REMI)转化的致病性诱变  

Mutation for Pathogenicity of Magnaporthe grisea by Restriction Enzyme Mediated Integration (REMI)

作　　者：张文荟[1] 周益军[1] 范永坚[1] 程兆榜[1] 吴淑华[1] 

机构地区：[1]江苏省农业科学院植物保护研究所,南京210014

出　　处：《南京师大学报（自然科学版）》2002年第2期17-21,共5页Journal of Nanjing Normal University(Natural Science Edition)

基　　金：国家自然科学基金 (39870 4 2 9);江苏省农科院基金 (6 11980 4 )

摘　　要：应用提高真菌转化率的技术———限制酶介导整合 (restrictionenzymemediatedintegration ,REMI)技术转化稻瘟病菌株Magnaporthegrisea131原生质体 ,在限制酶HindIII,KpnI或SacI介导下转化效率分别提高 6 .5、10、3.7倍 .通过 30 0 μg/mL潮霉素筛选获得 10 0 0个以上转化子 .用突变体接种水稻鉴别品种 ,发现其中M2 5和M36与M131比较其致病性发生了改变 .另外还发现产孢缺陷型突变体和培养性状发生改变的突变菌株 .repA new method called REMI (Restriction Enzyme Mediated Integration) has been used to transform Magnaporthe grisea 131 with plasmid pUCATPH.the linearized plasmid pUCATPH was transformed in the presence of additional restriction enzyme HindⅢ,KpnⅠ or SacⅠ.This greatly enhanced transformation efficiency to 6.5?10?3.7 times respectively.More than 1000 transformants were screened with 300?μg/mL Hygromycin B.Then they were cultured and inoculated on in vitro rice leaves for analysis of pathogenicity.Only two REMI transformants with pathogenic mutation on rice variety Aichi Asahi were thought as the results of REMI event.The mutants showed different fingerprints from the original isolate M131 by rep PCR analysis.It was deduced that mutation may involved in the function of inactivation of avr gene.sporulation defected and different culture phenotypical mutants were also found.

关 键 词：稻瘟病菌 限制酶介导整合 REMI 致病性诱变 产孢缺陷型突变体 病害防治 

分 类 号：S435.111.4[农业科学—农业昆虫与害虫防治]