• Korean Journal of Plant Resources
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• v.27 no.6
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• pp.680-686
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• 2014

Acanthopanacis Cortex has been used for oriental medicinal purposes in Asian countries especially in Korea and China. In the Korean Pharmacopeia, the cortexes of the dried roots, stems and branches of all species in Eleutherococcus and Eleutherococcus sessiliflorus are known as 'Ogapi'. Mostly the cortexes of E. gracilistylus roots and E.senticosus roots were used as 'Ogapi' in China and Japan, respectively. Therefore, the purpose of this study was to determine and compare the molecular authentication of Korean 'Ogapi' by using the ribosomal internal transcribed spacer (ITS) region. The ITS region has the highest possibility of effective and successful identification for the widest variety of molecular authentication. The ITS region was targeted for molecular analysis with Single nucleotide polymorphisms (SNPs) specific for morphologically similar to E. gracilistylus, E. senticosus, E. sessiliflorus from their adulterant, moreover, E. sieboldianus were detected within sequence data. Thus, based on these SNP sites, specific primers were designed and multiplex PCR analysis were conducted for molecular authentication of four plants (E. gracilistylus, E. senticosus, E. sessiliflorus, and E. sieboldianus). The findings of results indicated that ITS region might be established multiplex-PCR analysis systems and hence were proved to be an effective tools for molecular evaluation and comparison of 'Ogapi' with other plants.

• The Korea Journal of Herbology
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• v.29 no.6
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• pp.35-43
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• 2014

Objectives : Official Arisaematis Rhizoma is described only three species, Arisaema amurnse, Arisaema erubescens, and Arisaema heterophyllum, in national Pharmacopoeia. However, other Arisaema species, Arisaema ringens, Arisaema takesimense and Arisaema serratum, also have been distributed as an inauthentic Arisaematis Rhizoma in the herbal market. To develop a reliable molecular authentication method for Arisaematis Rhizoma in species level, we analyzed DNA barcode regions using six Arisaema species. Methods : Thirty-eight samples of six Arisaema plants species (A. amurense, A. amurense f. serratum, A. heterophyllum, A. takesimense, and A. serratum) were collected from different habitate and nucleotide sequences of DNA barcode regions (rDNA-ITS, matK, and rbcL gene) were analyzed after PCR amplification. The species-specific sequences and phylogenetic relations were estimated using entire sequences of three DNA barcodes based on the analysis of ClastalW and UPGMA, respectively. Results : The comparative analysis of DNA barcode sequences were revealed inter-species specific nucleotides to distinguish the medicinal plant of Arisaema Rhizoma in species levels excluding between A. amurense and its subspecies (A. amurense f. serratum) and A. takesimense and A. serratum, respectively. However, we obtained sequence differences enough to discriminate authentic and inauthentic Arisaematis Rhizoma. Therefore, we suggest that these SNP type molecular genetic markers were an reliable method avaliable to identify official herbal medicines. Conclusions : These marker nucleotides could be useful to identify the official herbal medicines by providing definitive information that can identify original medicinal plant and distinguish from inauthentic adulterants and substitutes.

• Journal of Life Science
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• v.14 no.3
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• pp.425-428
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• 2004

Molecular authentication and genetic polymorphism of Korean ginseng cultivars and accessions were investigated using ISSR (inter-simple sequence repeat amplification) markers. Five primers among 56 produced clear and reproducible DNA fragments among seven cultivars and accessions. A total of 43 bands ranging from 250 bp to 1,700 bp from five primers were scored. Average number of bands per primer was 8.6 and only nine bands were polymorphic across the six Panax ginseng from Korea. Especially Chunpoong cultivar exhibited the highest level of polymorphism, whereas other accessions did not showed almost any polymorphism. Consequently, these ISSR markers will be available to differentiate Chunpoong cultivar from other major Korean ginseng cultivars and accessions, such as Yunpoong, Hwangsukjong and Jakyungjong, at the DNA level.

• The Korea Journal of Herbology
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• v.21 no.3
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• pp.91-95
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• 2006

Objectives : This study was performed to determine if an antler could be identified as one of the Cervus species by phylogenetic analysis, which was used to assess genetic authentication. Methods : The DNAs of an antler were extracted, amplified by PCR, and sequenced. The DNAs of an antler were identified by Phylogenetic analysis. Phylogenetic analysis was made using MEGA software (Molecular Evolutionary Genetics Analysis, 3.1) Results : By phylogenetic analysis an antler was identified as Cervus elaphus nelsoni not as Cervus elaphus sibericus. This work showed that authentication can efficiently be performed by phylogenetic analysis. Conclusion : These results suggest that phylogenetic analysis might be able to provide the authentication of Cervus species.

• Plant Biotechnology Reports
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• v.4 no.1
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• pp.1-7
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• 2010

Definitive identification of original plant species is important for standardizing herbal medicine. The herbal medicines Cynanchi Wilfordii Radix (Baekshuoh in Korean and Beishuwu in Chinese) and Polygoni Multiflori Radix (Hashuoh in Korean and Heshuwu in Chinese) are often misidentified in the Korean herbal market due to morphological similarities and similar names. Therefore, we developed a reliable molecular marker for the identification of Cynanchi Wilfordii Radix and Polygoni Multiflori Radix. We used random amplified polymorphic DNA (RAPD) analysis of three plant species, Polygoni multiflorum, Cynanchum wilfordii, and Cynanchum auriculatum, to obtain several species-specific RAPD amplicons. From nucleotide sequences of these RAPD amplicons, we developed six sequence characterized amplification region (SCAR) markers for distinguishing Polygoni Multiflori Radix and Cynanchi Wilfordii Radix. Furthermore, we established SCAR markers for the simultaneous discrimination of the three species within a single reaction by using multiplex-PCR. These SCAR markers can be used for efficient and rapid authentication of these closely related species, and will be useful for preventing the distribution of adulterants.

• The Korea Journal of Herbology
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• v.28 no.6
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• pp.53-58
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• 2013

Objectives : Pinelliae Tuber has been used as a typical unauthentic herbal medicines. Due to the morphological similarity between Pinelliae Tuber and adulterants, the correct authentication is very difficult. Therefore, we introduced DNA barcode to establish a powerful tool for the authentication of Pinelliae Tuner from adulterants. Methods : To obtain DNA barcode regions, genomic DNA was extracted from nineteen specimens of Pinellia ternata, Pinellia pedatisecta, Pinellia tripartita, and Typhonium flagelliforme, and matK and rbcL genes were amplified. For identification of species specific sequences and analysis phylogenetic relationship, a comparative analysis were performed by the ClastalW and UPGMA based on entire sequences of matK and rbcL genes, respectively. Results : In comparison of two DNA barcode sequences, we elucidated the phylogenetic relationship showing distinct four groups depending on species and identified 40 and 20 species specific nucleotides enough to distinguish each species from matK and rbcL gene, respectively. The sequence differences at the corresponding positions were avaliable genetic marker nulceotides to discriminate the correct species among analyzed four species. These results indicated that phylogentic and comparative analysis of matK and rbcL genes are useful genetic markers to authenticate Pinelliae Tubers. Conclusions : The marker nucleotides enough to distinguish P. ternata, P. tripatrita, P. peditisecta, and T. flagelliform, were observed at 40 positions in matK gene and 20 positions in rbcL gene sequence, respectively. These differences can be used to authenticate Pinelliae Tuber from adulterants as well as discriminate each four species.

• Korean Journal of Plant Resources
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• v.26 no.3
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• pp.397-402
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• 2013

Morus Folium (Sang-yeop in Korean) is one of the most important Oriental medicinal plants. In Korea, both M. alba and M. cathayana are regarded as the botanical sources for Morus Folium. In order to discriminate M. alba and M. cathayana from their adulterant, M. tricuspidata, mitochondrial NADH dehydrogenase subunit 7 (nad7) intron 2 region was targeted for molecular analysis with universal primers. DNA polymorphisms, including SNP sites, insertions, and deletions, were detected among these three species sequencing data. Based on these DNA polymorphisms, specific primers were designed for the three species respectively. Multiplex PCR was conducted for molecular authentication of M. alba, M. cathayana, and M. tricuspidata with specific primers. The present results indicate that it is possible to identify Morus Folium from its adulterant using mitochondrial nad7 intron 2 region. The established multiplex-PCR system was proved to be effective for identification of Morus Folium. The results indicate that mitochondrial introns can be used for inter-specific polymorphic study, and the described method can be applied for molecular identification of medicinal materials.

• Natural Product Sciences
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• v.21 no.3
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• pp.185-191
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• 2015

Comparative molecular analysis has been frequently adopted for the authentication of herbal medicines as well as the identification of botanical origins. Roots and rhizomes of the family Umbelliferae have been used as traditional herbal medicines to relieve various symptoms such as inflammation, neuralgia and paralysis in countries of East Asia. Since most herbal medicines of the Umbelliferae roots or rhizomes are generally supplied in the form of dried slices, morphological examination does not often provide sufficient evidence to identify the botanical origin. Using species-specific probes developed by the comparative analysis of nrDNA ITS sequences, a DNA chip was developed to identify herbal medicines for 13 species in the Umbelliferae. The developed DNA Chip proves its potential as a rapid, sensitive and effective tool for authenticating herbal medicines in future.

• 한국약용작물학회:학술대회논문집
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• 2010.10a
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• pp.213-214
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• 2010

• 한국약용작물학회:학술대회논문집
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• 2010.10a
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• pp.215-216
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• 2010