四引物扩增受阻突变体系PCR检测水稻白叶枯病抗性基因xa5的优化与应用  

Optimization and Utilization of the Detection for Bacterial BlightResistance Gene xa5 Molecular Marker in Rice withTetra-primer ARMS PCR

作　　者：何开碧 武帅民 尤一航 李瑶[1,2] 王平 印汉[1,2] 夏志辉 HE Kaibi;WU Shuaimin;YOU Yihang;LI Yao;WANG Ping;YIN Han;XIA Zhihui(School of Breeding and Multiplication(Sanya Institute of Breeding and Multiplication),Hainan University,Sanya Hainan 572025,China;School of Tropical Agriculture and Forestry,Hainan University,Danzhou Hainan 571737,China)

机构地区：[1]海南大学南繁学院(三亚南繁研究院),海南三亚572025 [2]海南大学热带农林学院,海南儋州571737

出　　处：《种子》2025年第3期222-226,共5页Seed

基　　金：三亚崖州湾科技城科技专项(SCKJ-JYRC-2023-16);海南崖州湾种子实验室(B23YQ1512);中国种子集团有限公司“揭榜挂帅”项目(B23CQ15CP)。

摘　　要：xa5是水稻抗白叶枯病的隐性基因,其与显性感病等位基因Xa5的功能差异源于Xa5基因开放阅读框116号和117号的TC碱基被替换为AG。为开发稳定且易于识别的共显性功能标记以辅助分子育种,基于四引物扩增受阻突变体系PCR(Tetra-primer ARMS-PCR)原理,设计了六组四引物组合。通过优化引物设计及反应体系,筛选出可在1%琼脂糖凝胶电泳中清晰区分xa5纯合子(400/157 bp)、Xa5纯合子(400/289 bp)及杂合子(400/289/157 bp)的共显性标记(组合T5)。该标记经F_(2)群体验证符合孟德尔分离规律(χ^(2)=0.93),且抗病表型与基因型一致。xa5 is a recessive gene for resistance to rice(Oryzasativa L.)bacterial blight.The main functional difference of Xa5 is that the TC bases at positions 116 and 117 of the molecular breeding for rice resistance to bacterial blight.In order to develop an easily identifiable and stable functional marker,based on the principle of Tetra Primer Amplification Refractory Mutation System PCR(TPARMS PCR),six sets of tetra-primer combinations were designed.By optimizing primer design and reaction system,a co-dominant marker(combined T5)was selected that could clearly distinguish xa5 homozygote(400/157 bp),Xa5 homozygote(400/289 bp)and heterozygote(400/289/157 bp)in 1%agarose gel electrophoresis.The marker was verified by F_(2) population to conform to Mendelian isolation rule(χ^(2)=0.93),and the disease resistance phenotype was consistent with the genotype.

关 键 词：白叶枯病 xa5 四引物扩增受阻突变体系PCR 功能标记 

分 类 号：S511[农业科学—作物学]