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KMID : 1145720150030020073
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Korean Herbal Medicine Informatics
2015 Volume.3 No. 2 p.73 ~ p.80
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Identification of Marker Nucleotides for the Molecular Authentication of Paeoniae Radix Based on the Analysis of psbA-trnH Sequences
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Kim Wook-Jin
Ji Yun-Ui
Moon Byeong-Cheol
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Abstract
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Purpose : The original plant species of Paeoniae Radix is differentially described in the national pharmacopoeia of Korea, China and Japan. To develop the reliable method for the authentication of four Paeonia species, P. lactiflora, P. japonica, Paeonia veitchii and P. suffruticosa, we analyzed psbA-trnH DNA barcode region and identified species-specific marker nucleotides.
Materials and Methods : Plant materials were collected from different geographic regions in Korea and China, and psbA-trnH DNA barcode region were amplified using psbA3-f and trnHf-05 primer set. The DNA sequences were analyzed and aligned using the Bio-Edit program. The multiple alignment of each samples were also analyzed with the Bio-Edit program in order to calculate the genetic distance among the samples using unweight pair group method with arithmetic averages (UPGMA) anlysis.
Results : PCR amplification rate of 12 samples used in this study were 100% for psbA- trnH region. In comparison of psbA-trnH sequences among four Paeonia species, we identified 1, 2, 2, and 16 species specific nucleotides enough to distinguish each species, respectively. These species-specific sequences could provide useful genetic marker nucleotides to discriminate the precise species among the analyzed four Paeonia species.
Conclusion : We analyzed psbA-trnH region to confirm the availability of molecular authentication for Paeoninae Radix on species levels using four Paeonia species. As a result, we amplified PCR products of psbA-trnH region from all of the 12 samples and obtained total 21 marker nucleotides enough to distinguish each four Paeonia species. These marker nucleotides are useful genetic tool for the standardization of Paeonae Radix.
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KEYWORD
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Marker Nucleotide, Molecular Authentication, Paeonia species, Paeoniae Radix, psbA-trnH
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