A simple and rapid technique for the authentication of the ginseng cultivar, Yunpoong, using an SNP marker in a large sample of ginseng leaves |
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Authors: | Sun Hua Wang Hong-Tao Kwon Woo-Saeng Kim Yeon-Ju In Jun-Gyo Yang Deok-Chun |
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Affiliation: | a Department of Oriental Medicinal Material & Processing, College of Life Science, Kyung Hee University, 1 Seocheon-dong, Giheung-gu, Yongin-si, Gyeonggi-do 446-701, Republic of Koreab The State Key Laboratory of Plant Cell and Chromosome Engineering, National Centre for Plant Gene Research, Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Beichen West Road 1, Chaoyang District, Beijing 100101, China |
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Abstract: | 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. |
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Keywords: | SNP, Single nucleotide polymorphism EST, Expressed sequence tag GAPDH, Glyceraldehyde 3-phosphate dehydrogenase PCR, Polymerase chain reaction ISSR, Inter-simple sequence repeat PCR-RFLP, Polymerase chain reactions of restriction fragment length polymorphisms ARMS-PCR, Amplification refractory mutation system-PCR EST-derived PCR, Expressed sequence tag derived PCR InDel, Insertions and deletions |
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