稻瘟病广谱抗性基因Pigm特异性分子标记的开发和应用  

Development and Verification of Specific Molecular Markers for Pigm Gene Associated with Broad-spectrum Resistance to Rice Blast

作　　者：陈涛[1,2] 孙旭超 张善磊 梁文化 周丽慧[1] 赵庆勇[1] 姚姝[1] 赵凌[1] 赵春芳[1] 朱镇[1] 张亚东[1] 王才林[1] CHEN Tao;SUN Xuchao;ZHANG Shanlei;LIANG Wenhua;ZHOU Lihui;ZHAO Qingyong;YAO Shu;ZHAO Ling;ZHAO Chunfang;ZHU Zhen;ZHANG Yadong;WANG Cailin(Institute of Food Crops,Jiangsu Academy of Agricultural Sciences,Jiangsu High Quality Rice R&D Center/Nanjing Branch of China National Center for Rice Improvement,Nanjing 210014,China;Jiangsu Co-Innovation Center for Modern Production Technology of Grain Crops,Yangzhou University,Yangzhou 225009,China)

机构地区：[1]江苏省农业科学院粮食作物研究所/江苏省优质水稻工程技术研究中心/国家水稻改良中心南京分中心,南京210014 [2]扬州大学江苏省粮食作物现代产业技术协同创新中心,江苏扬州225009

出　　处：《中国水稻科学》2020年第1期28-36,共9页Chinese Journal of Rice Science

基　　金：国家重点研发计划重点专项(2017YFD0100305);江苏省重点研发计划资助项目(BE2018357);江苏省农业重大新品种创制项目(PZCZ201703);现代农业产业技术体系建设专项资金资助项目(CARS-01-62);江苏省农业科学院粮食作物研究所科研基金资助项目(LZS17-7)

摘　　要：【目的】来自籼稻品种谷梅4号的Pigm是一个对稻瘟病菌具有广谱和持久抗性的重要基因。为有效提高Pigm基因的选择效率,有必要开发具有特异性的共显性分子标记进行辅助育种。【方法】本研究根据谷梅4号Pigm位点存在的特异性核苷酸变异,利用Tetra-primerARMS-PCR和KASP两种不同的基因分型技术开发出分子标记T-Pigm和K-Pigm,对不同品种(品系)以及淮稻9号/谷梅4号的F2分离群体进行基因型检测,并结合穗颈瘟人工接种鉴定,对标记的准确性进行评价。【结果】序列比对分析表明,谷梅4号Pigm位点中Pigm-Nbs2基因起始密码子上游515 bp处存在一个特殊的单核苷酸变异。利用已开发的两种类型的分子标记能够有效区分3种不同的基因型,且基因型与穗颈瘟人工接种鉴定结果完全一致。【结论】利用分子标记T-Pigm和K-Pigm可以实现对Pigm基因型快速、准确的检测,加快抗稻瘟病水稻新品种的选育进程。【Objective】Pigm from an indica rice variety Gumei 4,is one of the most important genes with broad-spectrum and durable resistance to rice blast.It is very necessary to improve its selection efficiency by developing specific and co-dominant markers in breeding.【Method】The molecular markers T-Pigm and K-Pigm were developing with Tetra-primer ARMS-PCR and KASP technique,based on the specific nucleotide mutation in Pigm locus.Then,the consistency of the genotype and phenotype was evaluated in different varieties and the F2 population derived from Huaidao 9/Gumei 4 by artificial inoculation methods of panicle blast.【Result】The sequence alignment analysis indicated that a specific SNP was at-515 bp of the Pigm-Nbs2 gene initiation codon in Gumei 4.The three different genotypes of Pigm locus could be distinguished by the Tetra-primer ARMS-PCR and KASP markers,which exactly conformed to the results of artificial inoculation.【Conclusion】T-Pigm and K-Pigm markers could be used to detect the genotype of Pigm gene rapidly and accurately,and improved the selection efficiency of blast resistant rice varieties in breeding.

关 键 词：稻瘟病 Pigm 特异性标记 四引物扩增受阻突变体系PCR 竞争性等位基因特异性PCR 

分 类 号：S435.111.4[农业科学—农业昆虫与害虫防治] S511.034[农业科学—植物保护]