利用SLAF-seq结合BSA方法发掘大豆种皮色相关基因

种皮色不仅是一个形态标记,还是一种重要的进化性状,与大豆商品性、营养价值以及抗性关系密切。本研究以黄色种皮大豆品系BMY及其褐色种皮衍生品系BMZ为亲本,从F2群体中分别挑选出黄色种皮和褐色种皮的植株各9株,构建了两个极端性状混合池DNA文库,借助SLAF-seq和BSA技术,挖掘大豆种皮色性状相关候选基因。对于26121个高质量SNP位点,利用SNP-index方法和ED方法进行综合分析,获得8个与目标性状紧密关联的候选区域,分别位于第5、11、12、19和20染色体,包含620个编码基因。对这620个基因进行进一步的功能注释,NR、Swiss-Prot、GO、KEGG和COG数据库分别注释到600、518、533、133和256个基因,其中存在非同义突变基因4个,分别为Glyma05g005600、Glyma05g009700、Glyma12g006100和Glyma12g047300。候选区域内编码基因的深度注释有助于功能基因的进一步分离,为大豆种皮色性状相关基因的精细定位及克隆提供理论依据。

Mapping of Soybean Genes Related to Seed-coat Color Using SLAF-seq and BSA Methods

Seed-coat color is not only a morphological marker,but also an important evolutionary trait.Seed-coat color is closely related to soybean commodity,nutritional value and resistance.In this study,9 plants with yellow seed-coat and 9 plants with brown seed-coat were selected from the F2 population using a yellow seed-coat soybean line BMY and its brown seed-coat derived line BMZ as parents.The DNA libraries of two extreme trait(yellow and brown seed-coat colors)pools were constructed.Candidate genes were identified related to soybean seed-coat color traits using SLAF-seq and BSA techniques.For 26121 high-quality SNP loci,eight candidate regions were obtained closely related to the target traits by both SNP-index and ED methods.They were located on chromosomes 5,11,12,19 and 20,respectively,and contained 620 coding genes.Furthermore,600,518,533,133 and 256 genes were annotated by NR,Swiss-Prot,GO,KEGG and COG databases,respectively.Among them,there were 4 non-synonymous mutation genes identified,including Glyma05 g005600,Glyma05 g009700,Glyma12 g-006100 and Glyma12 g047300.Functional annotations of these candidate genes are helpful for further gene isolation and provide theoretical basis for fine mapping and cloning of the genes related to seed-coat color traits in soybean.