广谱抗稻瘟病基因PigmR的KASP标记开发及应用
收稿日期: 2024-09-08
修回日期: 2025-02-25
网络出版日期: 2025-05-21
基金资助
江苏省种业振兴揭榜挂帅项目[JBGS（2021）039];江苏省重点研发计划（现代农业）资助项目(BE2021374);国家自然科学基金面上项目(42071023);生物育种钟山实验室项目(BM2022008-03);生物育种钟山实验室项目(1201);生物育种钟山实验室项目(1002);江苏省自然科学基金资助项目(BK20212010)
Development and Application of KASP Marker for Broad-Spectrum Resistance Gene PigmR to Rice Blast
Received date: 2024-09-08
Revised date: 2025-02-25
Online published: 2025-05-21
【目的】广谱抗稻瘟病基因PigmR对水稻抗稻瘟病遗传改良具有重要育种价值。根据其编码区变异位点开发竞争性等位基因特异性PCR(Kompetitive allele specific，PCR KASP)标记，有利于提升PigmR育种利用的效率。【方法】通过序列比对明确PigmR基因编码区存在单核苷酸变异，开发KASP分子标记PigmR-K，通过对水稻品种和F2分离群体进行基因型分析，结合稻瘟病表型鉴定，并开展分子标记辅助选择育种，对标记的精准性和高效性进行验证。【结果】根据PigmR基因编码区序列第2503位存在特异性单核苷酸A>C变异，建立基于KASP的PigmR精准高效基因分型技术体系，通过对159份水稻品种、270份F2群体单株以及247份BC1F3群体单株进行基因型检测，PigmR-K能够高效区分供试水稻材料的纯合抗病、纯合感病和杂合基因型，并与人工接种穗颈瘟症状一致。【结论】利用PigmR基因编码区变异位点开发的KASP标记PigmR-K具有精准高效等优势，有利于稻瘟病抗性遗传改良。
李文奇, 许扬, 王芳权, 朱建平, 陶亚军, 李霞, 范方军, 蒋彦婕, 陈智慧, 杨杰 . 广谱抗稻瘟病基因PigmR的KASP标记开发及应用[J]. 中国水稻科学, 2025 , 39(3) : 365 -372 . DOI: 10.16819/j.1001-7216.2025.240104
【Objective】The broad-spectrum resistance gene PigmR has important breeding value for the genetic improvement of rice blast resistance. Developing kompetitive allele specific PCR (KASP) markers based on its coding region variation sites is beneficial for improving the accuracy and efficiency of PigmR utilization in breeding.【Method】This study identified single nucleotide variations in the coding region of PigmR through sequence alignment, developed the KASP molecular marker PigmR-K, conducted genotype analysis using rice varieties and F2 segregating populations, combined these results with rice blast symptom identification, and carried out molecular marker assisted selection breeding to verify the precision and efficiency of this marker.【Result】Based on an A-to-C mutation at position 2503 in the PigmR gene coding region sequence, a precise and efficient PigmR genotyping technology system based on KASP markers was established. The genotyping of 159 rice varieties, 270 F2 individuals, and 247 BC1F3 population lines demonstrated that PigmR-K reliably distinguished the homozygous resistant, homozygous susceptible and heterozygous genotypes of PigmR. These results were completely consistent with disease symptoms from artificial panicle blast inoculation.【Conclusion】The KASP marker PigmR-K developed based on the variation site of on the coding region has advantages such as precision and efficiency, which is beneficial for genetic improvement of rice blast resistance.
Key words： rice; broad-spectrum resistance; rice blast; PigmR gene; KASP; breeding efficiency
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