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Abstract
Panax ginseng is one of the most valuable medicinal plants in the Orient. The low level of genetic variation has limited the application of molecular markers for cultivar authentication and marker-assisted selection in cultivated ginseng. To exploit DNA polymorphism within ginseng cultivars, ginseng expressed sequence tags (ESTs) were searched against the potential intron polymorphism (PIP) database to predict the positions of introns. Intron-flanking primers were then designed in conserved exon regions and used to amplify across the more variable introns. Sequencing results showed that single nucleotide polymorphisms (SNPs), as well as indels, were detected in four EST-derived introns, and SNP markers specific to “Gopoong” and “K-1” were first reported in this study. Based on cultivar-specific SNP sites, allele-specific polymerase chain reaction (PCR) was conducted and proved to be effective for the authentication of ginseng cultivars. Additionally, the combination of a simple NaOH-Tris DNA isolation method and real-time allele-specific PCR assay enabled the high throughput selection of cultivars from ginseng fields. The established real-time allele-specific PCR assay should be applied to molecular authentication and marker assisted selection of P. ginseng cultivars, and the EST intron-targeting strategy will provide a potential approach for marker development in species without whole genomic DNA sequence information.
Highlights
Panax ginseng is one of the most valuable medicinal plants in the Orient
The results suggest that expressed sequence tags (ESTs) intron-targeting strategy could meet the needs of marker development for ginseng cultivars
Together with the single nucleotide polymorphisms (SNPs) markers we developed for “Gumpoong”, “Chunpoong”, and “Yunpoong” in previous studies, our established real-time allele-specific polymerase chain reaction (PCR) method can provide an effective authentication system, as well as a pure seed supply system, for P. ginseng cultivars
Summary
Panax ginseng is one of the most valuable medicinal plants in the Orient. The pharmacological effects of P. ginseng have been studied extensively, including improved brain function, enhanced immune system function, pain-alleviating effects, anti-tumor activity, regulated blood pressure, as well as anti-oxidative and anti-aging effects [1]. Ten cultivars including “Yunpoong”, “Gopoong”, “Sunpoong”, “Gumpoong”, “Chunpoong”, “K-1”, “Sunwon”, “Sunweon”, “Sunhyang”, and “Chungsun” are widely cultivated because they have superior agricultural traits [2]. These cultivars show higher yields and qualities, they are frequently mixed-cultivated with local landraces in ginseng fields, mainly due to the lack of a reliable method for authentication of ginseng cultivars. Ginseng products made from different cultivars and mixed seeds are sold in the market This impedes the management of ginseng cultivation, and affects the quality control of ginseng products
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