在水稻的杂种优势利用中,两系法因为不需要保持系、不育系,育性由单一核基因控制、配组不受恢保关系制约等优点而更受青睐.tms5类型的两系不育系在目前的两系杂交稻中占有绝对主导的地位,针对该基因的功能分子标记辅助育种将为两系法的应用提供一个更加准确和高效的方法.本研究将tms5基因的功能变异开发成基于高分辨率熔解曲线分析(high resolution melting analysis, HRM)检测的特异性分子标记,并对各种类型亲本进行了 HRM分型检测及测序验证.结果表明,该标记能够准确区分tms5基因的各种功能变异,且与测序结果完全吻合.将各种变异亲本与突变体'广占63S'杂交,并用所开发的功能分子标记对其相应的后代植株进行检测,发现该标记能够很好地区分各种类型亲本与突变体的F1代个体及后续的BC1F1和BC3F2代植株.因此,所开发的功能性分子标记可以用于tms5基因的资源鉴定和分子标记辅助改良,从而为两系不育系的准确高效分子育种提供技术支持.“,”In the utilization of heterosis in rice, two-line hybrid is preferred due to its no need of the maintainer line, sterile line, fertility is controlled by a single nuclear gene, and no restriction on combination by the restoring and maintaining relationships. The tms5-type two-line sterile line occupies an absolutely dominant position in the two-line hybrid rice at present. The functional marker assisted breeding of this gene will provide a more accurate and efficient method for the application of the two-line hybrid. In this study, we developed the functional variation of tms5 gene into a specific molecular marker based on HRM detection, and applied HRM genotyping and sequencing in all kinds of parents. The experimental results showed that the marker could accurately detect various functional variations of the tms gene, which was fully agreement with the sequencing results. The parents of the various variants were hybridized with the mutant 'Guangzhan 63S', and the corresponding progeny plants were estimated by the functional molecular marker. The marker accurately differentiated the F1 individuals generated by parents with various variations at TMS5 and the mutants, as well as the subsequent BC1F1 and BC3F2 generation plants. Therefore, the developed functional marker could be used for resource identification and marker-assisted improvement of tms5 gene, providing technical support for the accurate and efficient molecular breeding of the two-line sterile lines.