洋葱雄性不育恢复位点Ms座位的SSR标记开发及其应用

The Development and Application of SSR Markers Linked to Male Sterility Restoration Site Ms Locus in Allium cepa L.

本研究旨在开发稳定可靠的洋葱育性位点分子标记,并将其应用于育种实践,筛选育种系,从而缩短育种周期,节省育种成本,加速洋葱杂交种的培育进程。以洋葱育性恢复Ms座位的AFLP序列为基础,采用序列比对分析、PCR检测和表型分析等方法,开发、鉴定、应用了WH-SSR-1分子标记。结果表明:WH-SSR-1标记与Ms位点紧密连锁,Ms位点基因型为纯合显性MsMs时,有1条102 bp的扩增带;纯合隐性msms时,有1条99 bp的扩增带;杂合Msms时,有102 bp和99 bp两条扩增带。32份洋葱材料的验证结果证实了Ms座位的标记类型与基因型完全相符。随后从4个OP群体中直接获得了2个配套的不育系与保持系,‘吊玉’和‘天正红玉’因未检测到保持株或不育株,无法简单实现配套。WH-SSR-1标记与Ms位点紧密连锁,能够将其用于育种系筛选的育种实践,从OP群体中直接选择保持株和不育株,同时实现两系配套。

This study aims to develop a stable and reliable molecular marker of onion fertility locus and apply it to breeding practice for screening breeding lines, which will further shorten the breeding cycle, save breeding costs, and accelerate the cultivation process of onion hybrids. Based on the AFLP sequences of onion fertility restoration Ms locus, WH-SSR-1 molecular marker was developed, identified and applied in breeding practice by sequence alignment analysis, PCR detection and phenotype analysis. The results showed that WH-SSR-1 marker was closely linked to Ms locus. When Ms locus genotype was homozygous dominant MsMs, there was one 102 bp band;when Ms locus genotype was homozygous recessive msms, there was only one 99 bp band;when Ms locus genotype was hybridized Msms, there were two bands, 102 bp band and 99 bp band,respectively. The validation results of 32 onion materials confirmed that the marker type of Ms locus was completely consistent with the genotype. Subsequently, two matching maintainer and sterile lines were directly developed from four OP populations, but‘Diaoyu’and‘Tianzhenghongyu’could not be easily matched because no maintainer or sterile individuals were detected. The WH-SSR-1 marker is closely linked to Ms locus and can be used in the breeding practice. The maintainer and sterile individuals can be directly selected from OP population, and can be matched with each other and form counterpart breeding lines at the same time.