稻瘟病广谱抗性基因Pigm特异性分子标记的开发和应用
收稿日期: 2019-07-12
修回日期: 2019-11-20
网络出版日期: 2020-01-10
基金资助
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)
Development and Verification of Specific Molecular Markers for Pigm Gene Associated with Broad-spectrum Resistance to Rice Blast
Received date: 2019-07-12
Revised date: 2019-11-20
Online published: 2020-01-10
【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-primer ARMS-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。
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
陈涛, 孙旭超, 张善磊, 梁文化, 周丽慧, 赵庆勇, 姚姝, 赵凌, 赵春芳, 朱镇, 张亚东, 王才林 . 稻瘟病广谱抗性基因Pigm特异性分子标记的开发和应用[J]. 中国水稻科学, 2020 , 34(1) : 28 -36 . DOI: 10.16819/j.1001-7216.2020.9082
【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.
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