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根癌农杆菌介导的稻瘟病菌近等菌系的构建与初步应用*
引用本文:何余勇,周惠萍,吴兴兴,吴毅歆,王志远,毛自朝,何月秋.根癌农杆菌介导的稻瘟病菌近等菌系的构建与初步应用*[J].云南农业大学学报,2010,25(5):603-611.
作者姓名:何余勇  周惠萍  吴兴兴  吴毅歆  王志远  毛自朝  何月秋
作者单位:[1]云南农业大学,农业生物多样性与病虫害控制教育部重点实验室,云南昆明650201 [2]云南农业大学农学与生物技术学院,云南昆明650201
基金项目:国家行业计划,国家"863"计划项目,农业部"948"项目,国家行业计划项目 
摘    要: 利用根癌农杆菌C58C1介导转化1株致病力极弱的野生型稻瘟病菌菌株CY2,成功地获得3000多个转化子,混合接种于携带25个不同抗病单基因的水稻近等基因系上,获得4株单胞致病突变体;继代培养和PCR检测表明,这些突变体均为T-DNA插入CY2的后代,而非自发突变或外源菌株污染。将4株突变体与实验室已保存的20株致病突变体分别回接水稻近等基因系,推断其携带的致病基因类型,结果表明突变体分别针对13个抗性单基因的18个单基因系发生了突变,其中针对Pi-a,Pik-s,Pi-sh单基因系表现出较高的侵染频率。由于这些突变体均来自同一个菌株,仅致病性存在差异,属于近等菌系,利用其分析出102个水稻品种中的34个品种的抗病基因组成,其中,抗病基因Pik-s,Pi-a,Pi-sh,Pi-19(t)分布频率较高,分别达70.59%,61.76%,52.94%和44.12%,且带有这类基因的糯稻品种多于杂交水稻。

关 键 词:稻瘟病菌  近等菌系  根癌农杆菌  T-DNA转化  抗病基因鉴定
收稿时间:2010-04-06

Agrobacterium tumefaciens-Mediated Construction and Preliminary Application of Near-isogenic Isolates of Magnaporthe oryzae
HE Yu-yong,ZHOU Hui-ping,WU Xing-xing,WU Yi-xin,WANG Zhi-yuan,MAO Zi-chao,HE Yue-qiu,.Agrobacterium tumefaciens-Mediated Construction and Preliminary Application of Near-isogenic Isolates of Magnaporthe oryzae[J].Journal of Yunnan Agricultural University,2010,25(5):603-611.
Authors:HE Yu-yong  ZHOU Hui-ping  WU Xing-xing  WU Yi-xin  WANG Zhi-yuan  MAO Zi-chao  HE Yue-qiu  
Institution:1.Laboratory of Agricultural Biodiversity and Pests Control, Ministry of Education, Yunnan Agricultural University, Kunming 650201, China;2.College of Agriculture and Biotechnology, Yunnan Agricultural University, Kunming 650201, China
Abstract:Magnaporthe oryzae CY2 with weak pathogenicity to rice was transformed by Agrobacterium tumefaciens C58C1 mediation, and more than 3000 transformants were obtained, then 4 pathogenic mutants were screened out among them by inoculating onto the rice near isogenic lines with 25 different resistance genes. The results of secondary cultures on the medium with hygromycin B and PCR amplification indicated the mutants were CY2s transformants by the insertion of T-DNA but not contaminated isolates. Then, 4 pathogenic mutants plus other 20 pathogenic mutants in the laboratory were inoculated back onto the rice near isogenic lines to investigate their different virulence genes, showing those mutants were pathogenic to rice lines with 13 different resistance genes, especially to the rice with Pi-a, Pik-s, Pi-shgenes. Due to the same origin and only pathogenicity difference, those mutants belong to near isogenic isolates. They could detect rice resistance genes according to the gene for gene hypothesis. By the isolates, we investigated the resistance gene of 34 among 102 rice cultivars. The genes Pik-s, Pi-a, Pi-sh andPi-19(t) were more common with 70.59%, 61.76%, 52.94% and 44.12% frequencies than others among 34 cultivars and more in glutinous than in hybrid rice.
Keywords:Magnaporthe oryzae  near isogenic isolate  Agrobacterium tumefacien  transformation  resistance gene identification
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