萝卜Ogura-CMS育性恢复基因Rfo的高通量SNP标记开发及其应用

利用萝卜Ogura-CMS保持系和恢复系育性恢复基因Rfo的序列,开发了基于高通量竞争性等位基因特异性PCR(KASP)分型技术的SNP标记:Rfo-SNP1。利用该标记对24个萝卜品种的304个单株进行了育性恢复基因Rfo的基因型鉴定,其中289株获得基因分型结果,占整个群体的95.07%。从分型结果看,育性恢复基因Rfo的基因型为rfrf的有110株(占36.18%),基因型为Rf Rf和Rfrf的分别为156株(占51.32%)和23株(占7.57%)。在12个品种中获得基因型为rfrf的单株,可作为保持系并用于不育系转育。利用PCR-RFLP标记和回交群体单株育性鉴定,进一步证实Rfo-SNP1标记不但能够实现高通量检测,而且能够实现萝卜Ogura-CMS育性恢复基因Rfo基因型的精准鉴定。目前利用该方法已成功转育了6套稳定遗传的雄性不育系及保持系。

Development and Application of High-throughput SNP Markers for Ogura-CMS Fertility Restorer Gene in Radish

Based on the nucleotide sequences of Rfo related to the Ogura-CMS fertility restorer gene in radish,the SNP (Single nucleotide polymorphism) marker named Rfo-SNP1 was developed by use of KASP (kompetitive allele specific PCR) technology.The genotypes of fertility restorer gene in 24 radish varieties including 304 plants were identified using the Rfo-SNP1 marker.The results showed that 289 plants were screened out accounting for 95.07％ of the whole population.And 110 plants had the recessive homozygous genotype (rfrf) taking up 36.18％.The dominant homozygous genotype (RfRf) and the heterozygous genotype (Rfrf) appeared respectively in 156 plants and 23 plants.In 23 fertile varieties,12 varieties had the rfrfgenotype,which implied that the 12 varieties might be the candidates of maintainer lines for breeding sterile lines.The accuracy of Rfo-SNP1 markers was further verified by PCR-RFLP and backcross populations.Six male sterile lines and maintainer lines of radish have been developed through this procedure.