Data Sheet 2_An initial exploration of core collection construction and DNA fingerprinting in Elymus sibiricus L. using SNP markers.xlsx

Elymus sibiricus L., an excellent forage and ecological restoration grass, plays a key role in grassland ecological construction and the sustainable development of animal husbandry. In China, the wild germplasm resources of E. sibiricus are abundant, and they are shaped by similar and contrasting climatic conditions to form distinct populations, which enrich the genetic diversity of E. sibiricus. To more comprehensively aggregate E. sibiricus germplasm resources at a lower cost and to more accurately utilize its genetic variation, this study conducted a preliminary exploration of core germplasm collections and fingerprinting of E. sibiricus using single nucleotide polymorphism (SNP) markers. By combining multiple evaluation measures with weighted processing, we successfully identified 36 materials from 90 wild E. sibiricus samples to serve as a core collection. Genetic diversity assessments, allele evaluations, and principal component analyses of the 36 core germplasm samples all indicate that these 36 samples accurately and comprehensively represent the genetic diversity of all 90 E. sibiricus germplasm accessions. Additionally, we identified 290 SNP loci from among the high-quality SNP loci generated by whole-genome sequencing of the 90 E. sibiricus samples as candidate markers. Of these, 52 SNP loci were selected as core markers for DNA fingerprinting of E. sibiricus. Using kompetitive allele-specific PCR (KASP) technology, we also performed population origin identification for 60 wild E. sibiricus germplasm accessions based on these core markers. The core SNP markers screened in this study were able to accurately distinguish between E. sibiricus germplasms from the Qinghai–Tibet Plateau and those from elsewhere. This study not only provides a reference for the continued collection and identification of E. sibiricus germplasm resources but also offers a scientific basis for their conservation and utilization.

西伯利亚披碱草（Elymus sibiricus L.）是兼具优良饲用价值与生态修复功能的草本植物，在草地生态建设与畜牧业可持续发展领域发挥关键作用。我国境内的西伯利亚披碱草野生种质资源十分丰富，受相似与差异化气候条件的共同塑造，形成了多个遗传分化显著的种群，进一步丰富了该物种的遗传多样性。为以更低成本更全面地聚合西伯利亚披碱草种质资源、更精准地开发利用其遗传变异，本研究借助单核苷酸多态性（single nucleotide polymorphism, SNP）标记技术，针对西伯利亚披碱草开展核心种质库构建与DNA指纹图谱构建的初步探索。本研究结合多种评价方法并进行加权处理，从90份野生西伯利亚披碱草样本中筛选出36份材料作为核心种质库。对该36份核心种质样本开展的遗传多样性评价、等位基因分析及主成分分析结果均显示，其能够准确且全面地代表90份西伯利亚披碱草种质资源的整体遗传多样性。此外，本研究从90份样本全基因组测序获得的高质量SNP位点中筛选出290个作为候选标记，其中52个SNP位点被选为西伯利亚披碱草DNA指纹图谱构建的核心标记。本研究借助竞争性等位基因特异性PCR（kompetitive allele-specific PCR, KASP）技术，基于上述核心标记对60份野生西伯利亚披碱草种质资源开展种群来源鉴定，结果显示本研究筛选得到的核心SNP标记可准确区分青藏高原与其他区域的西伯利亚披碱草种质资源。本研究不仅为西伯利亚披碱草种质资源的后续收集与鉴定工作提供了参考依据，同时也为该物种种质资源的保护与利用奠定了科学基础。