Division of Forest Genetic Resources, Korea Forest Research Institute, Suwon, 441-847, Korea.
Mol Biol Rep. 2012 Jan;39(1):729-36. doi: 10.1007/s11033-011-0792-4. Epub 2011 May 15.
Cleaved amplified polymorphic sequence (CAPS) marker system using mitochondrial consensus primers was applied for molecular identification of Korean ginseng cultivars (Panax ginseng). Initially, a total of 34 primers were tested to six Korean ginseng cultivars and two foreign Panax species, P. quinquefolius and P. notoginseng. In the polymerase chain reaction (PCR) amplification results, four primers (mt7, mt11, mt13, and mt18) generated co-dominant polymorphic banding patterns discriminating the Korean ginseng cultivars from P. quinquefolius and P. notoginseng. In the CAPS analysis results, the majority of the cleaved PCR products also yielded additional latent polymorphisms between the Korean ginseng cultivars and two foreign Panax species. Specific latent CAPS polymorphisms for cultivar Gopoong and Chunpoong were detected from internal region amplified with mt9 primer by treating HinfI and Tsp509I endonucleases, respectively. Sequencing analysis revealed that the length of amplified region of Korean ginseng cultivars was 2,179 bp, and those of P. quinquefolius and P. notoginseng were 2,178 and 2,185 bp, respectively. Blast search revealed that the amplified region was a mitochondrial cytochrome oxidase subunit 2 (cox2) gene intron II region. Nineteen single nucleotide polymorphisms (SNP) including each specific SNP for Gopoong and Chunpoong, and three insertion and deletion (InDel) polymorphisms were detected by sequence alignment. The CAPS markers developed in this study, which are specific to Gopoong and Chunpoong, and between the Korean ginseng cultivars and two foreign Panax species, will serve as a practical and reliable tool for their identification, purity maintenance, and selection of candidate lines and cultivars.
采用线粒体通用引物的切割扩增多态性序列 (CAPS) 标记系统被应用于朝鲜人参品种的分子鉴定。最初，对 6 个朝鲜人参品种和 2 个外国人参属植物，即西洋参和三七，共测试了 34 个引物。在聚合酶链反应 (PCR) 扩增结果中，4 个引物 (mt7、mt11、mt13 和 mt18) 产生了区分朝鲜人参品种与西洋参和三七的共显性多态性带型。在 CAPS 分析结果中，大多数切割的 PCR 产物在朝鲜人参品种和两个外国人参属植物之间也产生了额外的潜在多态性。通过分别用 HindIII 和 Tsp509I 内切酶处理 mt9 引物扩增的内部区域，检测到 Gopoong 和 Chunpoong 品种的特异性潜在 CAPS 多态性。测序分析表明，朝鲜人参品种的扩增区域长度为 2179 bp，西洋参和三七的扩增区域长度分别为 2178 和 2185 bp。Blast 搜索表明，扩增区域是线粒体细胞色素氧化酶亚单位 2 (cox2) 基因内含子 II 区。通过序列比对，共检测到 19 个单核苷酸多态性 (SNP)，包括每个 Gopoong 和 Chunpoong 的特异性 SNP，以及 3 个插入和缺失 (InDel) 多态性。本研究开发的特异性针对 Gopoong 和 Chunpoong 以及朝鲜人参品种与两个外国人参属植物的 CAPS 标记将成为其鉴定、纯度保持以及候选品系和品种选择的实用可靠工具。
