Combined use of molecular markers and high resolution melting (HRM) to assess chromosome dosage in potato hybrids

In plants, the most widely used cytological techniques to assess parental genome contributions are based on in situ hybridization (FISH and GISH), but they are time-consuming and need specific expertise and equipment. Recent advances in genomics and molecular biology have made PCR-based markers a straightforward, affordable technique for chromosome typing. Here, we describe the development of a molecular assay that uses single-copy conserved ortholog set II (COSII)-based single nucleotide polymorphisms (SNPs) and the high-resolution melting (HRM) technique to assess the chromosome dosage of interspecific hybrids between a Solanum phureja–S. tuberosum diploid (2n = 2x = 24) hybrid and its wild relative S. commersonii. Screening and analysis of 45 COSII marker sequences allowed S. commersonii-specific SNPs to be identified for all 12 chromosomes. Combining the HRM technique with the establishment of synthetic DNA hybrids, SNP markers were successfully used to predict the expected parental...在植物研究中，用于评估亲本基因组贡献的最常用细胞学技术基于原位杂交（in situ hybridization）体系，具体包括荧光原位杂交（FISH）与基因组原位杂交（GISH），但此类技术耗时较长，且需具备专业操作技能与专用实验设备。近年来基因组学与分子生物学的研究进展，使基于聚合酶链式反应（Polymerase Chain Reaction, PCR）的分子标记成为一种操作简便、成本可控的染色体分型技术。本研究详述了一种分子检测方法的开发流程：该方法利用基于单拷贝保守同源基因集II（COSII）的单核苷酸多态性（SNPs）结合高分辨率熔解（HRM）技术，以评估Solanum phureja–S. tuberosum二倍体杂种（2n = 2x = 24）与其近缘野生种S. commersonii之间的种间杂种的染色体剂量。通过对45条COSII标记序列的筛选与分析，我们在全部12条染色体上均鉴定出了S. commersonii特异性的SNPs位点。结合HRM技术与人工合成DNA杂种的构建，本研究成功利用SNP标记实现了对预期亲本...