Department of Oriental Medicinal Material & Processing, Kyung Hee University, Yongin-si, Republic of Korea.
Gene. 2011 Nov 1;487(1):75-9. doi: 10.1016/j.gene.2011.05.021. Epub 2011 Aug 9.
Yunpoong is an important Korean ginseng (Panax ginseng C. A. Meyer) cultivar, but no molecular marker has been available to identify Yunpoong from other cultivars. In this study, we developed a single nucleotide polymorphism (SNP) marker for Yunpoong based on analysis of expressed sequence tags (ESTs) in an exon region of the glyceraldehyde 3-phosphate dehydrogenase (GAPDH) gene. This SNP marker had high specificity to authenticate Yunpoong in twelve different main ginseng cultivars. For application of the molecular marker, a rapid identification method was established based on the NaOH-Tris method and real-time polymerase chain reaction (PCR) in order to ensure more efficiency in the cultivar selection. The biggest feature of the NaOH-Tris method was that it made the extraction of DNA very simple and rapid in young leaf tissues. We only spent 1 min to extract DNA and directly used it to do PCR. In this report, the conventional DNA extraction method was used to develop molecular marker process, and the NaOH-Tris method was applied in screening large numbers of cultivars. Moreover, the greatest advantage of the real-time PCR compared with traditional PCR, is time saving and high efficiency. Thus, this strategy provides a rapid and reliable method for the specific identification of Yunpoong in a large number of samples.
云鹏是一种重要的高丽参（Panax ginseng C. A. Meyer）品种，但目前还没有分子标记物可用于将其与其他品种区分开来。本研究基于甘油醛 3-磷酸脱氢酶（GAPDH）基因外显子区的表达序列标签（EST）分析，为云鹏开发了一个单核苷酸多态性（SNP）标记。该 SNP 标记在 12 种不同的主要人参品种中对云鹏具有高度特异性。为了应用该分子标记，我们基于 NaOH-Tris 法和实时聚合酶链反应（PCR）建立了一种快速鉴定方法，以确保在品种选择方面更加高效。NaOH-Tris 法的最大特点是，它使得幼叶组织中的 DNA 提取非常简单和快速。我们仅花费 1 分钟即可提取 DNA，并直接将其用于 PCR。在本报告中，常规 DNA 提取方法用于开发分子标记过程，NaOH-Tris 法用于筛选大量品种。此外，与传统 PCR 相比，实时 PCR 的最大优势是节省时间和提高效率。因此，该策略为大量样品中云鹏的特异性鉴定提供了一种快速可靠的方法。
