【摘要】 基于Super-GBS基因分型（super-genotyping-by-sequencing）技术开展柞蚕单核苷酸多态性标记（single nucleotide polymorphism, SNP）位点挖掘研究，旨在开发性状关联SNP分子标记，为柞蚕种质资源评价鉴定、遗传图谱构建、QTL定位提供数据支撑。以白体色小白蚕为母本、黄体色H04为父本，获得BC₁M群体（110个）、F₁（3个）、P₁（1个）、P₂（1个）总计115个柞蚕材料，利用PstⅠ-HF/MspⅠ双酶切基因组构建Super-GBS文库并测序，使用BWA软件将过滤后的序列数据比对到柞蚕参考基因组，采用GATK软件开发SNP标记，使用SnpEff软件注释SNP位点及R语言分析遗传结构。测序共产生序列数据量为106.39 Gb,过滤后的平均读长为0.89 Gb, Q30测序质量值平均为90.11%,比对率平均为98.48%。共得到有效SNP位点141 100个，主要位于基因间隔区、内含子区、基因下游、上游，其中C/T、A/G变异类型最多，转换与颠换的比例为1.296 187∶1。SNP位点变异主要为同义突变和错义突变，产生修饰（MODIFIER）影响。主成分与聚类分析将115个样品分为4组（P₁、P₂、F₁、BC₁M）,反映了样品的遗传背景及亲缘关系，群体遗传分化指数F_ST在0.000 377 7～0.8272间，群体遗传距离DR分布在0.000 4～1.755 6,F_ST与DR均表现出明显的遗传背景上的聚类。结果表明，Super-GBS技术能够获得大量柞蚕SNP位点信息，可用于SNP标记开发，且开发的SNP标记能够对115个样品的亲缘关系、体色演变进行解释，为今后柞蚕种质资源评价与鉴定、分子标记辅助育种工作奠定基础。更多还原

【Abstract】 The study of single nucleotide polymorphism（SNP） loci data mining in tussah（Antheraea pernyi） based on Super-GBS（genotyping-by-sequencing） technology is aimed at the development of trait-associated SNP molecular markers, which will provide data support for the evaluation and identification of Antheraea pernyi germplasm resources, construction of genetic maps, and QTL localization. Using Xiaobaican（white body color） as the mother and H04（yellow body color） as the father, we obtained a total of 115 materials including BC₁M population（110）, F₁（3）, P₁（1） and P₂（1）. We built a Super-GBS library and sequenced it by using PstⅠ-HF/MspⅠ double enzyme digestion of genome and aligned the filtered sequence data to the Antheraea pernyi reference genome by using software BWA. The GATK software was used to develop SNP markers, SnpEff software was used to annotate SNP sites and R language was used to analyze the genetic structure. The total amount of sequence data was 106.39 Gb, with a filtered average read length of 0.89 Gb, an average Q30 sequencing quality value of 90.11%, and an average comparison rate of 98.48%. We obtained a total of 141 100 valid SNP loci, mainly in the intergenic region, intron region, downstream, and upstream, with the most types of C/T and A/G variants, and the Ts/Tv ratio of 1.296 187∶ 1. The SNP loci mutations were mainly silent and missense mutation, which produced the effect of modification（MODIFIER）. According to the results of principal component and cluster analysis, the 115 samples were divided into four groups（P₁, P₂, F₁, BC₁M）, reflecting the genetic background and relatives of the samples. The population genetic differentiation index F_ST ranged from 0.000 377 7 to 0.8272, and the population genetic distance DR ranged from 0.000 4 to 1.755 6, both showing obvious clustering on genetic background. The results showed that a large amount of information on Antheraea pernyi SNP loci could be obtained by using Super-GBS technology, which could be used for the development of SNP markers, these markers could explain the genetic relationship and evolution of body colors of 115 samples, laying foundation for the evaluation and identification of Antheraea pernyi germplasm resources and molecular marker-assisted breeding in the future.更多还原