基于SLAF-seq技术的苦瓜白粉病SNP分子标记开发

以高抗白粉病苦瓜MC18和高感白粉病MC105杂交构建遗传分离群体,利用SLAF-seq技术进行深度测序,开发与抗白粉病性状紧密关联的SNP分子标记。结果表明:测序共获得各样品的有效reads为10.48 Mb,获得高质量的SLAF标签91 856个,其中多态性SLAF标签5 502个,多态性比例为5.99%。通过对多态性SLAF标签进行关联分析,筛选出与苦瓜白粉病抗病性状紧密关联的SNP位点共29个。根据关联分析结果 ,通过四引物扩增受阻突变体系PCR技术进行SNP位点多态性的验证,其中Marker4542、Marker11188分子标记特异性强、灵敏度高、稳定性好,建立一套简便快捷的苦瓜SNP分型方法。该方法可对苦瓜育种材料进行分子标记辅助选择,以进一步提高苦瓜抗病育种效率。 

Development of SNP Molecular Markers for Powdery Mildew in Bitter Gourd Using SLAF-seq Technique

MC18 ( highly powdery mildew -resistant), MC105 ( highly powdery mildew -sensitive) and  the  F2  segregating population were used as the experimental materials for sequencing by specific-locus amplified fragment sequencing (SLAF-seq), a high-throughput reduced-representation genotyping technology. In this study, a total of 10.48 Mb reads were obtained, and 91 856 high quality SLAFs were produced, of which 5 502 were polymorphisms. Finally, we developed 29 specific fragments of powdery mildew in bitter gourd using SLAF-seq technique, and Tetra-primer ARMS-PCR technology was used to validate SNP genetic typing of bitter gourd, and Marker4542, Marker11188 molecular markers detected could be closely  linked  to  the  genes  corresponding  to  the  target  traits. The results would lay a good foundation for bitter gourd MAS resistance breeding.