水稻野败型三系杂交种纯度检测功能标记的开发与应用OA

[Objective]Developing stable,efficient,and reliable markers for purity testing of three-line hybrid seeds is crucial for current hybrid rice seed production.[Method]Based on the specific sequences of the wild abortive-type sterility gene WA352,the major fertility-restoring gene Rf4,and the internal reference gene Actin,multiple single-gene detection markers were designed.PCR amplification was then performed by optimizing the primer mixture ratio.[Result]A mixture of primers WA352-2,Rf4-3,and Actin-3 in a ratio of 1:11:3 enabled simultaneous detection of the genotypes of WA352,Rf4,and Actin in a single PCR reaction,allowing differentiation among wild abortive-type CMS lines,restorer lines,maintainer lines,and hybrids.[Conclusion]The detection method established in this study is simple to perform and yields stable and reliable results.It can not only identify genotypes in wild-abortive three-line rice but also serve as an effective tool for molecular marker-assisted breeding.By improving the accuracy of purity assessment during breeding,this method provides strong support for achieving high-yield and high-quality rice breeding objectives in China.