• 제목/요약/키워드: Molecular authentication

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Molecular Authentication of Acanthopanacis Cortex by Multiplex-PCR Analysis Tools

  • Kim, Min-Kyeoung;Jang, Gyu-Hwan;Yang, Deok-Chun;Lee, Sanghun;Lee, Hee-Nyeong;Jin, Chi-Gyu
    • 한국자원식물학회지
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    • 제27권6호
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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.

천남성(天南星) 유전자 감별을 위한 DNA 바코드 분석 및 Marker Nucleotide 발굴 (Identification of Marker Nucleotides for the Molecular Authentication of Arisaematis Rhizoma Based on the DNA Barcode Sequences)

  • 김욱진;이영미;지윤의;강영민;최고야;김호경;문병철
    • 대한본초학회지
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    • 제29권6호
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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.

ISSRs 마크에 의한 고려 인삼의 분자적 인증과 유전적 다형현상 (Molecular Authentication and Genetic Polymorphism of Korean Ginseng (Panax ginseng C. A. Meyer) by Inter-Simple Sequence Repeats (ISSRs) Markers)

  • Bang, Kyong-Hwan;Lee, Sung-Woo;Hyun, Dong-Yun;Cho, Joon-Hyeong;Cha, Seon-Woo;Seong, Nak-Sul;Huh, Man-Kyu
    • 생명과학회지
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    • 제14권3호
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    • pp.425-428
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    • 2004
  • ISSR마크를 사용하여 고려 인삼의 품종 및 계통간 분자적 인증과 유전적 다형현상을 조사하였다. 56개의 ISSR 프라이머 중 5개가 일곱 품종 및 계통간 명확하고 재현성이 높은 DNA분절을 나타내는 최적 프라이머로 선택되었다. 전체 43밴드는 250 bp - 1,700 bp의 분자량을 가지며 프라이머당 8.6개의 밴드를 나타내었다. 고려 인삼에서 다형현상 정도는 20.9%였다. 특히 천풍 품종이 가장 높은 다형현상을 나타낸 반면 다른 품종은 거의 다형현상을 나타내지 않았다. 결론적으로 DNA수준에서 ISSR마크로 천풍이 다른 고려 인삼의 품종 및 계통인 연풍, 황숙종, 자경종과 구분에 이용될 수 있음이 판명되었다.

Ensuring Consumer Safety: Molecular Authentication of Eurycoma longifolia Derivative Products in the Wood Science and Technology Industry

  • Arida SUSILOWATI;Henti Hendalastuti RACHMAT;Kusumadewi Sri YULITA;Asep HIDAYAT;Susila SUSILA;Nawwall ARROFAHA;Irsyad KAMAL;Fifi Gus DWIYANTI
    • Journal of the Korean Wood Science and Technology
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    • 제52권4호
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    • pp.343-362
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    • 2024
  • Eurycoma longifolia (pasak bumi) is a popular medicinal plant in Indonesia and is widely used in various products. Its high economic value has caused illegal harvesting and product falsification. Using molecular techniques, the authentication and traceability of E. longifolia derivatives can be controlled to ensure consumer safety. Therefore, this study aimed to authenticate the products and derivatives of E. longifolia (pasak bumi) produced, marketed, and consumed in Indonesia using molecular identification techniques. Genomic DNA from 37 leaf samples collected from the Sumatran mainland and the Riau Islands and six E. longifolia products were amplified and sequenced using trnL-trnF and internal transcribed spacer (ITS) regions. The results revealed that all leaf samples were indeed E. longifolia based on the markers used, with the six products, only the herbal tea product (sample code TCPB) was most likely derived from E. longifolia based on the two regions, suggesting that not all products labelled as E. longifolia in the market are authentic. The results also indicated that several other plants species are used as substitutes or adulterants, including Simaba spp., Simarouba spp., Homalolepis spp., Vernonia gigantea, Elephantopus scaber, Gymnanthemum amygdalinum, Cyanthillium spp., Potentilla lineata, Ailanthus altissima, Geijera paniculata, Hannoa chlorantha, and Dalbergia spp. Klebsiella pneumoniae bacteria were also identified in this study on the outer wooden cup of E. longifolia products. Therefore, this molecular approach is effective in identifying the authenticity of E. longifolia products, with trnL-trnF and ITS as the recommended DNA markers.

Cervus 종의 Phylogenetic analysis에 의한 판별 (Authentication of Cervus Species by Phylogenetic analysis)

  • 서정철;김민정;이찬;김명규;이정수;최강덕;임강현
    • 대한본초학회지
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    • 제21권3호
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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.

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Rapid molecular authentication of three medicinal plant species, Cynanchum wilfordii, Cynanchum auriculatum, and Polygonum multiflorum (Fallopia multiflorum), by the development of RAPD-derived SCAR markers and multiplex-PCR

  • Moon, Byeong-Cheol;Choo, Byung-Kil;Cheon, Myeong-Sook;Yoon, Tae-Sook;Ji, Yun-Ui;Kim, Bo-Bae;Lee, A-Young;Kim, Ho-Kyoung
    • Plant Biotechnology Reports
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    • 제4권1호
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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.

matK와 rbcL DNA 바코드 분석을 통한 반하(半夏) 및 반하(半夏) 유사 한약재 유전자 감별 (Molecular Authentication of Pinelliae Tuber from its adulterants by the analysis of DNA barcodes, matK and rbcL genes)

  • 이영미;문병철;지윤의;김욱진;김호경
    • 대한본초학회지
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    • 제28권6호
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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.

Molecular Authentication of Morus Folium Using Mitochondrial nad7 Intron 2 Region

  • Jin, Chi-Gyu;Kim, Min-Kyeung;Kim, Jin-Young;Sun, Myung-Suk;Kwon, Woo-Saeng;Yang, Deok-Chun
    • 한국자원식물학회지
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    • 제26권3호
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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.

The Application of DNA Chip Technology to Identify Herbal Medicines: an Example from the Family Umbelliferae

  • Kim, Pil-Ho;Park, Jisoo;Kim, Yeong Shik;Suh, Youngbae
    • Natural Product Sciences
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    • 제21권3호
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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.