Most satsuma mandarin (Citrus unshiu Marc.) cultivars are difficult to identify in the seedling stage based only on morphological traits. Therefore, simple polymerase chain reaction (PCR)-based single-nucleotide polymorphism (SNP) markers were developed to specifically and rapidly distinguish the ‘Haryejosaeng’ cultivar, which is generally supplied to breeders of other satsuma mandarin cultivars. SNP markers were verified using high-resolution melt (HRM)-specific primers. PCR was performed to distinguish ‘Haryejosaeng’ from eight other satsuma mandarin cultivars using six SNP markers (P1–P6) specific for ‘Haryejosaeng’, with one negative control SNP primer pair. The best results were obtained using three SNP markers (P1, P2, and P5). In the multiplex PCR, markers P1, P2, and P5 yielded 165-, 150-, and 526-base pair amplicons, respectively, in ‘Haryejosaeng’, distinguishing it from other satsuma mandarin cultivars. The selected SNP markers were validated by HRM with HRM-specific primers. The multiplex PCR with P1/P5 and P2/P5 also identified ‘Haryejosaeng’ obtained from a farm growing 17 different cultivars of satsuma mandarin. Specific SNP molecular markers were determined for accurately identifying the ‘Haryejosaeng’ cultivar by multiplex PCR to save the time and costs associated with its supply to breeders of satsuma mandarin.

大多数萨摩uma（Citrus unshiu Marc。）品种在苗期很难仅根据形态性状鉴定。因此，开发了基于简单聚合酶链反应（PCR）的单核苷酸多态性（SNP）标记，以特异性和快速地区分'Haryejosaeng'品种，该品种通常提供给其他萨摩普通话品种的育种者。使用高分辨率熔体（HRM）特异性引物验证了SNP标记。使用六个“ Haryejosaeng”特异的SNP标记（P1-P6）和一对阴性对照SNP引物进行PCR，以将“ Haryejosaeng”与其他八个萨摩普通话品种区分开。使用三个SNP标记（P1，P2和P5）可获得最佳结果。在多重PCR中，标记P1，P2和P5分别产生165、150和526个碱基对的扩增子，在“ Haryejosaeng”中，与其他萨摩普通话品种有所区别。选定的SNP标记已通过HRM使用HRM特异性引物进行了验证。用P1 / P5和P2 / P5进行的多重PCR还鉴定出了'Haryejosaeng'，这是从一个种植了17个不同萨摩uma普通品种的农场获得的。确定了特定的SNP分子标记，以通过多重PCR准确鉴定'Haryejosaeng'品种，以节省与向萨摩uma普通话提供给其育种者的时间和成本。