PDF(Pubmed)
Abstract：
Wine cultivars are available to growers in multiple clonal selections with agronomic and enological differences. Phenotypic differences between clones originated from somatic mutations that accrued over thousands of asexual propagation cycles. Genetic diversity between grape cultivars remains unexplored, and tools to discriminate unequivocally clones have been lacking. This study aimed to uncover genetic variations among a group of clonal selections of 4 important Vitis vinifera cultivars: Cabernet sauvignon, Sauvignon blanc, Chardonnay, and Merlot, and use this information to develop genetic markers to discriminate the clones of these cultivars. We sequenced with short-read sequencing technology the genomes of 18 clones, including biological replicates for a total of 46 genomes. Sequences were aligned to their respective cultivar\'s reference genome for variant calling. We used reference genomes of Cabernet sauvignon, Chardonnay, and Merlot and developed a de novo genome assembly of Sauvignon blanc using long-read sequencing. On average, 4 million variants were detected for each clone, with 74.2% being single nucleotide variants and 25.8% being small insertions or deletions (InDel). The frequency of these variants was consistent across all clones. From these variants, we validated 46 clonal markers using high-throughput amplicon sequencing for 77.7% of the evaluated clones, most of them small InDel. These results represent an advance in grapevine genotyping strategies and will benefit the viticulture industry for the characterization and identification of the plant material.
摘要：
葡萄酒品种可用于种植者的多种克隆选择，具有农艺和植物学差异。克隆之间的表型差异源于在数千个无性繁殖周期中产生的体细胞突变。葡萄品种之间的遗传多样性仍未被探索，一直缺乏明确区分克隆人的工具。这项研究旨在揭示四个重要葡萄品种的一组克隆选择之间的遗传变异：赤霞珠，长相思,霞多丽,和Merlot，并利用这些信息开发遗传标记来区分这些品种的克隆。我们用短读测序技术对18个克隆的基因组进行了测序，包括总共46个基因组的生物复制。将序列与各自品种的参考基因组进行比对，以进行变异识别。我们使用了赤霞珠的参考基因组,霞多丽,还有Merlot,并使用长读数测序开发了长相思长白的从头基因组组装。平均而言,每个克隆检测到4百万个变体，其中74.2%是单核苷酸变体,25.8%是小插入或缺失(InDel)。这些变体的频率在所有克隆中是一致的。从这些变体中，我们使用高通量扩增子测序验证了46个克隆标记的77.7%的评估克隆，他们中的大多数都是小InDel。这些结果代表了葡萄基因分型策略的进步，并将使葡萄种植行业有益于植物材料的表征和鉴定。
