摘要： 为了进一步定位Pis1基因，加快小麦的分子标记辅助育种工作，获得高质、高产、稳产的小麦品种。以川麦28（CM28）与其三雌蕊近等基因系CM28TP为研究材料，对CM28和CM28TP幼穗的3个阶段（幼穗长度为0.2~0.5 cm，0.5~0.7 cm，0.7~1.0 cm）进行转录组测序，然后通过GO数据库对变异位点所在的基因进行分类分析，并选取4个位于Pis1基因附近的SNP标记进行验证。结果表明，CM28TP和CM28幼穗3个阶段共有的SNP/InDel位点为5 310个，其中SNP位点5 024个，InDel位点286个。SNP位点中转换类型（63.33%）多于颠换类型（31.28%），两者的比值达到了2.02；InDel位点中插入类型（152个）多于缺失类型（134个）；SNP/InDel位点在A基因组上分布最多、其次是B基因组、D基因组上最少。对SNP/InDel所在的基因进行GO分类注释表明，生物学进程中基因的占比最高，其次是细胞组分和分子功能。SNP/InDel位点对蛋白质功能的影响预测表明，有36个变异位点会严重影响蛋白质功能，中度影响蛋白质功能的位点有1 279个。从Pis1基因的定位区间附近筛选了4个SNP位点进行PCR扩增和测序验证，发现这4个位点与RNA-Seq分析结果一致，这表明挖掘出的SNP/InDel位点是准确的。本研究丰富了小麦中的SNP/InDel标记，为小麦高密度遗传图谱构建、基因定位和分子标记辅助选择育种奠定了基础，同时开发出的4个位于Pis1基因附近的SNP标记，为图位克隆该基因提供了可能。

关键词: 三雌蕊小麦, 转录组, SNP标记, InDel标记

Abstract: In order to further locate the Pis1 gene, accelerate the molecular marker-assisted breeding of wheat, and obtain the wheat varieties with high quality, high yield and stable yield. Chuanmai 28(CM28) and its three pistils near-isogenic line CM28TP were used as research materials, the three stages of CM28 and CM28TP young spikes(spikelet length 0.2-0.5 cm, 0.5-0.7 cm, 0.7-1.0 cm) perform transcriptome sequencing, then, the GO database was used to classify and analyze the genes where the mutation site was located, and four SNP markers near the Pis1 gene were selected for verification. The results showed that there were 5 310 common SNP/InDel loci in CM28TP and CM28 young spikes at three stages, including 5 024 SNP loci and 286 InDel loci.In SNP sites, the conversion types(63.33%) were more than the transversion types(31.28%), and the ratio of the two types reached 2.02. There were more insert types(152) than deletion types(134) in InDel site. The SNP/InDel locus was most distributed in the A genome, followed by the B genome and the D genome was the least. The GO classification annotation of SNP/InDel genes showed that genes accounted for the highest proportion in biological processes, followed by cell components and molecular functions. The prediction of the impact of the SNP/InDel loci on protein function indicated that 36 mutant loci severely affected protein function, and 1 279 mutant loci moderately affected protein function. Four SNP sites were screened from the location interval of the Pis1 gene for PCR amplification and sequencing verification, and it was found that these four sites were consistent with the results of RNA-seq analysis, which indicated that the SNP/InDel sites excavated were accurate. This study enriched SNP/InDel markers in wheat and laid a foundation for high-density genetic map construction, gene mapping and marker-assisted selection breeding in wheat, at the same time, four SNP markers near the Pis1 gene were developed, which provided the possibility of map-based cloning of this gene.

Key words: Tripistil wheat, Transcriptome, SNP Marker, InDel Marker

中图分类号: 

• S512.03

• Q78