Establishment and application of a SNP molecular identification system in Grifola frondosa

Bin Diao, Zhixiang Xu, Min Liu, Guolin Zhang, Guangyuan Wang, Yinghao Zhang, Xuemei Tian
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Abstract

Germplasm resources of edible mushrooms are essential for the breeding of varieties with improved traits. Analysis of the genetic diversity of Grifola frondosa germplasm resources and clarification of the genetic relationships among strains can provide valuable information for the selection of breeding parents. A total of 829,488 high-quality SNP loci were screened from 2,125,382 SNPs obtained by sequencing 60 G. frondose. Phylogenetic analysis, PCA, and population structure analysis based on the high-quality SNPs showed that the 60 strains could be divided into five subgroups, and the clustering results were consistent with the geographical distributions of these strains. Based on high-quality SNP loci, a core collection containing 18 representative germplasm resources was constructed, and 1,473 Kompetitive Allele-Specific PCR markers were obtained. A total of 722 SNP markers in the exonic regions were screened using KASP-genotyping experiments, and 50 candidate SNP markers and 12 core SNP markers were obtained. Genetic fingerprints of G. frondosa germplasm resources were constructed based on the selected SNP markers; these fingerprints provide an accurate, rapid, convenient, and efficient method for the identification of G. frondosa germplasm resources. The results of this study have important implications for the preservation and utilization of G. frondosa germplasm resources and the identification of varieties.
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建立和应用草履虫 SNP 分子鉴定系统
食用菌种质资源对于培育具有改良性状的品种至关重要。分析双孢蘑菇种质资源的遗传多样性和阐明菌株间的遗传关系可为育种亲本的选择提供有价值的信息。通过对 60 份 G. frondose 的 2,125,382 个 SNPs 进行测序,共筛选出 829,488 个高质量 SNP 位点。基于高质量 SNP 的系统发育分析、PCA 和种群结构分析表明,60 个品系可分为 5 个亚群,聚类结果与这些品系的地理分布一致。基于高质量 SNP 位点,构建了包含 18 个代表性种质资源的核心收集,并获得了 1,473 个竞争性等位基因特异性 PCR 标记。利用 KASP 基因分型实验筛选了外显子区共 722 个 SNP 标记,获得了 50 个候选 SNP 标记和 12 个核心 SNP 标记。根据筛选出的 SNP 标记构建了 G. frondosa 种质资源的遗传指纹图谱;这些指纹图谱为 G. frondosa 种质资源的鉴定提供了准确、快速、便捷和高效的方法。该研究结果对保护和利用洋二仙草种质资源以及品种鉴定具有重要意义。
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