广东省侵染美香占2号的稻瘟病菌致病性及无毒基因变异分析  

Pathogenicity and Avirulence Genes Variation of Magnaporthe oryzae from a Rice Variety Meixiangzhan 2 in Guangdong Province

作　　者：汪文娟[1] 苏菁[1] 陈深[1] 杨健源[1] 陈凯玲[1] 冯爱卿[1] 汪聪颖[1] 封金奇 陈炳[1] 朱小源[1] WANG WenJuan;SU Jing;CHEN Shen;YANG JianYuan;CHEN KaiLing;FENG AiQing;WANG CongYing;FENG JinQi;CHEN Bing;ZHU XiaoYuan(Plant Protection Research Institute,Guangdong Academy of Agricultural Sciences/Guangdong Provincial Key Laboratory of High Technology for Plant Protection,Guangzhou 510640)

机构地区：[1]广东省农业科学院植物保护研究所/广东省植物保护新技术重点实验室,广州510640

出　　处：《中国农业科学》2022年第7期1346-1358,共13页Scientia Agricultura Sinica

基　　金：国家重点研发计划(2016YFD0300707);广东省自然科学基金(2020A1515011213);广东省农业科学院“优秀博士”人才引进项目(R2021YJ-YB3020);广东省农业科学院学科团队建设项目(202116TD);国家现代农业产业技术体系建设专项(CARS-01-35);广东省现代农业产业技术体系(2021KJ105);广州市科技计划(202002030001)。

摘　　要：【目的】分析侵染广东稻区优质品种美香占2号稻瘟病菌(Magnaporthe oryzae)的致病性及无毒基因变异特征,为美香占2号在不同稻区的合理布局提供参考依据。【方法】利用9个抗稻瘟病单基因系,对稻瘟病菌单孢分离菌株进行致病性测定;并利用8个稻瘟病菌已克隆无毒基因的功能性分子标记对2013—2018年不同年份、不同稻区采集的稻瘟病菌单孢分离菌株的DNA进行PCR扩增,通过琼脂糖凝胶电泳检测分析,选取不同带型和不同年份代表性菌株的相应无毒基因全长或CDS区域的PCR片段进行测序。测序结果与相应无毒基因的参考序列进行碱基序列的比较分析;并利用相应的水稻抗稻瘟病单基因系,对无毒基因不同变异类型的稻瘟病菌菌株进行致病性测定。根据稻瘟病菌两种交配型MAT1-1和MAT1-2型分子标记,检测侵染美香占2号的稻瘟病菌可能存在的交配型。【结果】致病性分析结果显示,2013-2018年采集自美香占2号品种的52个稻瘟病菌菌株对其中的4个单基因系IRBL9-W(Pi9)、IRBLzt-T(Piz-t)、NIL-e1(Pi50)和IRBLkh-K3(Pikh)均表现无毒;然而,侵染单基因系IRBLz-Fu(Piz)、IRBLkp-K60(Pikp)和IRBLi-F5(Pii)的有毒菌株频率维持在57%以上,且侵染单基因系IRBLz-Fu(Piz)的有毒菌株频率在不同年份间呈现显著性差异,表明美香占2号种植区田间稻瘟病菌群体中存在较高频率的avrPiz、avrPikp和avrPii毒性菌株。根据8个已克隆无毒基因的功能性分子标记检测,在52个供试菌株中均可检测到无毒基因AvrPiz-t、AvrPi9和AvrPik-CDE,但扩增不到Avr1-CO39、AvrPia和AvrPii,仅有4个菌株可检测到无毒基因AvrPita,有3个菌株可检测到无毒基因AvrPik-ABF。所测序7个菌株的AvrPi9编码区序列和AvrPiz-t包含启动子及编码区的序列与其各自无毒基因的序列完全一致;所测序7个菌株的AvrPik-CDE包含启动子及编码区的序列中,有4个菌株的序列与AvrPik无毒基因序�【Objective】The objective of this study is to analyze the pathogenicity and variation patterns of avirulence genes(Avr genes) genotype of Magnaporthe oryzae, which was collected from a high-quality rice variety Meixiangzhan 2 widely cultivated in Guangdong, and to provide a reference for the rational layout of Meixiangzhan 2 in different rice ecological areas.【Method】The pathogenicity of single-spore strains was determined using 9 M. oryzae monogenic differentials. The DNA of the single-spore M.oryzae strains collected from Meixiangzhan 2 in different rice areas and in different years from 2013 to 2018 was subjected to PCR amplification using the functional markers of 8 Avr genes. The PCR products of Avr genes with promotor sequences or CDS regions were analyzed by agarose gel electrophoresis and the PCR products of representative strains were sequenced, and their sequences were compared to corresponding Avr genes, respectively. The corresponding M. oryzae monogenic differentials were used to determine the pathogenicity of M. oryzae strains with different mutation types of the Avr genes. According to two mating types MAT1-1 and MAT1-2 molecular markers of M. oryzae, the possible mating type of M. oryzae strains isolated from Meixiangzhan 2was detected.【Result】The pathogenicity analysis with a set of monogenic differentials showed that 52 rice blast strains collected from Meixiangzhan 2 from 2013 to 2018 were avirulent to IRBL9-W(Pi9), IRBLzt-T(Piz-t), NIL-e1(Pi50) and IRBLkh-K3(Pikh).The tested strains showed high frequencies(>57%) of virulence to IRBLz-Fu(Piz), IRBLkp-K60(Pikp) and IRBLi-F5(Pii), and the frequency of virulence to IRBLz-Fu(Piz) showed significant differences in different years, suggesting that the M. oryzae population in the field of Meixiangzhan 2 planted area had a higher frequency of virulent strains of avrPiz, avrPikp and avr Pii. AvrPiz-t, AvrPi9and AvrPik-CDE fragments were almost present in all tested strains, but none of Avr1-CO39, AvrPia or AvrPii was amplified in any strain.

关 键 词：美香占2号 稻瘟病菌 无毒基因型 交配型 

分 类 号：S435.111.41[农业科学—农业昆虫与害虫防治]