• 제목/요약/키워드: Molecular Authentication

검색결과 48건 처리시간 0.025초

범용성 DNA 바코드(matK, rbcL) 분석을 통한 독활(獨活) 유전자 감별용 Marker Nucleotide 발굴 (Identification of Marker Nucleotides for the Molecular Authentication of Araliae Continentalis Radix Based on the Analysis of Universal DNA Barcode, matK and rbcL, Sequences)

  • 김욱진;양선규;최고야;문병철
    • 대한본초학회지
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    • 제31권5호
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    • pp.15-23
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    • 2016
  • Objectives : Araliae Continentalis Radix and Angelicae Pubescentis Radix have been used as the same medicinal name Korean and Chinese traditional medicines, respectively. The authentic Araliae Continentalis Radix is described only the root of Aralia continentalis in the Korean Pharmarcopoeia. However, the dried root of Angelica biserrata, Levisticum officinale, or Heracleum moellendorffii also has been distributed adulterants of Araliae Continentalis Radix. To develop a reliable method for identifying Araliae Continentalis Radix from adulterants, we carried out the analyses of universal DNA barcode sequences.Methods : Four plants species were collected from different habitate and nucleotide sequences of matK and rbcL were analyzed. The species-specific sequences and phylogenetic relationship were estimated using entire sequences of two DNA barcodes, respectively.Results : In comparative analysis of matK sequences, we were identified 104 positions of marker nucleotide for Ar. continentalis, 3 for An. biserrata, 4 for L. officinale and 8 for H. moellendorffii enough to distinguish individual species, respectively. Furthermore, we obtained marker nucleotides in rbcL at 42 positions for Ar. continentalis, 5 for An. biserrata and 2 for H. moellendorffii, but not for L. officinale. The phylogenetic tree of matK and rbcL were showed that all samples were clustered into four groups constituting homogeneous clades within the species.Conclusions : We confirmed that species-specific marker nucleotides of matK sequence provides distinct genetic information enough to identify four species. Therefore, we suggest that matK gene is useful DNA barcode for discriminating authentic Araliae Continentalis Radix from inauthentic adulterants.

Molecular Authentication of Magnoliae Flos Using Robust SNP Marker Base on trnL-F and ndhF Region

  • Kim, Min-Kyeoung;Noh, Jong-Hun;Yan, Deok-Chun;Lee, Sanghun;Lee, Hee-Nyeong;Jin, Chi-Gyu
    • 한국자원식물학회지
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    • 제28권3호
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    • pp.341-349
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    • 2015
  • Magnoliae Flos (Sini in Korean) is one of the most important oriental medicinal plants. In the Korean Herbal Pharmacopeia, the bud of the all species in Manolia denudate and Manolia genus were regarded as the botanical sources for ‘Sini’. Most the dried bud of Manolia denudata, Manolia biondii and Manolia sprengeri were used as ‘Xin-yi’ in China. Therefore, the purpose of this study was to determine and compare the ‘Magnolia’ species, four species including Manolia denudata, M. biondii, M. liliiflora and M. Kobus were analysis of sequencing data revealed DNA polymorphisms. The based on tRNA coding leucine/phenylalanine (trnL-F) and NADH-plastoquinone oxidoreductase subunit 5 (ndhF) sequences in chloroplast DNA. For the identification of ‘Magnolia’ species, polymerase chain reaction (PCR) analysis of chloroplast DNA regions such as ndhF have proven an appropriate method. A single nucleotide polymorphism (SNP) has been identified between genuine “Sini” and their fraudulent and misuse. Specific PCR primers were designed from this polymorphic site within the sequence data, and were used to detect true plants via multiplex PCR.

Rheological Properties of Dandelion Root Concentrates by Extraction Solvents

  • Lee, Ok-Hwan;Kang, Suk-Nam;Lee, Boo-Yong
    • Food Science and Biotechnology
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    • 제15권1호
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    • pp.33-38
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    • 2006
  • This study was performed to provide basic rheological data of dandelion root concentrates in order to predict their processing aptitude and usefulness as functional foods material. The hot water and 70% ethanol extracts of dandelion root were concentrated at 5, 20, and 50 Brix, and their static viscosity, dynamic viscosity, and Arrhenius plots were investigated. Almost all hot water concentrates showed the typical flow properties of a pseudoplastic fluid, but evaluation using the power law model indicated that the 70% ethanol concentrates showed a flow behavior close to a Newtonian fluid. The apparent viscosity of hot water and 70% ethanol concentrates decreased with increasing temperature. Yield stresses of hot water and 70% ethanol concentrates by Herschel-Bulkley model application were in the range of 0.026 - 1.368 Pa and 0.022 - 0.238 Pa, respectively. The effect of temperature and concentration on the apparent viscosity was examined by Arrhenius equation. The activation energies of hot water and 70% ethanol concentrates were in the range of $8.762-23.778{\times}10^3\;J/mol{\cdot}kg$ and $3.217-20.384{\times}10^3\;J/mol{\cdot}kg$ with increasing concentration, respectively. Storage (G') and loss (G") moduli were generally increased with increasing frequency. For the 70% ethanol concentrates, G" predominated over G' at all applied frequencies and so they showed the typical flow behavior of a low molecular solution. However, for the hot water concentrates, G' predominated over G" at more than 1.9 rad/sec (cross-over point) and so they showed the typical flow behavior of a macromolecular solution.

현삼속 식물의 종판별을 위한 Mitochondrial DNA의 염기서열 및 PCR-RFLP 분석 (Molecular Authentication of Scrophularia herbs by PCR-RFLP Based on rpl-5 Region of Mitochondrial DNA)

  • 이정훈;조익현;이제완;박춘근;방경환;김홍식;박충범
    • 한국약용작물학회지
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    • 제18권3호
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    • pp.173-179
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    • 2010
  • This study describes an efficient approach to the development of DNA markers for use in distinguishing the Scrophularia species that have been used as useful medicinal crops. In order to distinguish Scrophularia species, DNA sequences of rpl-5 region in mitochondrial DNA of Scrophularia species were analysed for detecting sequence variations, and the PCR-RFLP method was applied for developing practicable DNA marker patterns. Several DNA variations were detected by the sequence comparison of rpl-5 region among Scrophularia species. Genetic relationship analysis of Scrophularia species was carried out based on these DNA variations. DNA variations of rpl-5 region were revealed that it was significantly efficient in genetic relationship analysis of Scrophularia species. In addition, Scrophularia species tested in this study were completely discriminated by four polymorphic genotypes by PCR-RFLP combined with Tsp509 I (^AATT) restriction enzyme. Our results suggested that DNA sequence variations of rpl-5 region were sufficiently useful for genetic relationship analysis of Scrophularia species. Polymorphic genotypes by PCR-RFLP using the Tsp509 I enzyme will be useful for discrimination of Scrophularia species as a practicable DNA markers.

ITS 염기서열 기반 방기 신속 감별용 SCAR marker 개발 (Development SCAR marker for the rapid authenticaton of Sinomeni Caulis et Rhizoma based on ITS Sequences)

  • 김욱진;노수민;최고야;문병철
    • 대한본초학회지
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    • 제37권4호
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    • pp.9-16
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    • 2022
  • Objectives : In the Korean Pharmacopoeia 12th edition (KP 12) and the Korean Herbal Pharmacopoeia (KHP), two authentic herbal medicines are described, namely Bang-gi (Cheong-pung-deung) and Mok-bang-gi, respectively. In China, Bun-bang-gi is also used as herbal medicine. This study was conducted to develop a molecular authentication tool for distinguishing the three herbal medicine used as Bang-gi, which are Sinomeni Caulis et Rhizoma (Rhizome of Sinomenium acutum), Stephaniae Tetrandrae Radix (Root of Stephania terandra), and Cocculi Radix (Root of Cocculus trilobus). Methods : Twelve samples of three species (four samples of S. acutum, five samples of S. tetrandra, and three samples of C. trilobus) were collected from different habitats. The sequences of internal transcribed spacer (ITS) regions were obtained and comparatively analyzed to design the species-specific sequence characterized amplified region (SCAR) primers. The specificity of each pair of SCAR primers that amplified species-specific amplicon was evaluated for establishing the singleplex and multiplex PCR assay tools. Results : The singleplex SCAR markers show discriminability in C. acutum, S. tetrandra, and C. trilobus. These SCAR markers were also efficiently authenticated three species in the multiplex SCAR amplification using single PCR reaction. Furthermore, these PCR assay methods were applicable to authenticate dried herbal medicines distributed in the markets. Conclusions : The SCAR markers and PCR assay tools help discriminate the three herbal medicines used as Bang-gi at the species levels and provide a reliable genetic method to prevent the inauthentic distribution of these herbal medicines.

Microsatellite 마커를 이용한 딸기 품종의 DNA Profile Database 구축 (Construction of DNA Profile Data Base of Strawberry Cultivars Using Microsatellite Markers)

  • 홍지화;최근진;권용삼
    • 원예과학기술지
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    • 제32권6호
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    • pp.853-863
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    • 2014
  • 국내외에서 재배되고 있는 딸기 100품종의 DNA profile 데이터베이스를 구축하기 위하여 다형성이 높은 microsatellite 마커의 선정과 유전적 유사도 분석을 통한 품종식별력 검정 등에 대한 연구를 수행하였다. 딸기 21품종을 274개의 microsatellite 마커로 검정하여 반복 재현성이 높은 25개의 다형성이 높은 마커를 선정하였다. 이들 마커와 국내외에서 재배되고 있는 딸기 100품종을 검정하였을 때 마커당 평균 대립유전자수는 7.50개로 나타났고, 3-13개까지 다양한 분포를 나타내었다. PIC 값은 마커의 유전자형에 따라 0.333-0.841 범위에 속하였으며 평균값도 0.706으로 높게 나타났다. Microsatellite 마커의 대립유전자를 이용하여 딸기 100 품종에 대한 계통도를 작성하였을 때 품종 육성의 계보 및 육성 지역에 따라 7개의 그룹으로 크게 나누어졌으며 2품종을 제외한 98품종이 microsatellite 마커의 유전자형에 의해 식별이 되는 것으로 나타났다. 본 연구에서 얻어진 딸기 품종별 DNA profile 데이터베이스는 품종보호 출원 품종의 재배심사 및 품종진위성과 관련된 종자분쟁을 해결하는 수단으로 유용하게 활용될 수 있을 것이다.

The Complete Chloroplast Genome Sequence and Intra-Species Diversity of Rhus chinensis

  • Kim, Inseo;Park, Jee Young;Lee, Yun Sun;Joh, Ho Jun;Kang, Shin Jae;Murukarthick, Jayakodi;Lee, Hyun Oh;Hur, Young-Jin;Kim, Yong;Kim, Kyung Hoon;Lee, Sang-Choon;Yang, Tae-Jin
    • Plant Breeding and Biotechnology
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    • 제5권3호
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    • pp.243-251
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    • 2017
  • Rhus chinensis is a shrub widely distributed in Asia. It has been used for traditional medicine and ecological restoration. Here, we report the complete chloroplast genome sequence of two R. chinensis genotypes collected from China and Korea. The assembled chloroplast genome of Chinese R. chinensis is 149,094 bp long, consisting of a large single copy (97,246 bp), a small single copy (18,644 bp) and a pair of inverted repeats (16,602 bp). Gene annotation revealed 77 protein coding genes, 30 tRNA genes, and 4 rRNA genes. A phylogenomic analysis of the chloroplast genomes with 11 known complete chloroplast genomes clarified the relationship of R. chinensis with the other plant species in the Sapindales order. A comparative chloroplast genome analysis identified 170 SNPs and 85 InDels at intra-species level of R. chinensis between Chinese and Korean collections. Based on the sequence diversity between Korea and Chinese R. chinensis plants, we developed three DNA markers useful for genetic diversity and authentication system. The chloroplast genome information obtained in this study will contribute to enriching genetic resources and conservation of endemic Rhus species.

RAPD 분석을 통한 대황(大黃)과 종대황(種大黃) 감별용 SCAR 유전자 마커 개발 (Development of SCAR Markers for the Discrimination of Rhei Radix et Rhizoma and Rhei Undulatai Rhizoma based on the RAPD)

  • 문병철;이영미;천진미;이아영;윤태숙;전명숙;추병길;김호경
    • 대한본초학회지
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    • 제24권4호
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    • pp.115-120
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    • 2009
  • Objectives : Due to the morphological similarity and frequent occurrence of intermediate forms as well as morphological variations of aerial part, the correct identification between Rhei Radix et Rhizoma and Rhei Undulatai Rhizoma is very difficult. To develop a reliable method for correct identification and improving the quality standards of Rhei Radix et Rhizoma and Rhei Undulatai Rhizoma, we analyzed RAPD and developed SCAR marker. Methods : To amplify target DNA at the genomic level, 32 Operon 10-mer random primers were applied with four Rheum species, R. officinale, R. palmatum, R. tanguticum and R. undulatum. The nucleotide sequences were determined and species-specific primers were prepared depending on the species-specific RAPD amplicons after subcloned into the pGEM-Teasy vector. To develop the SCAR markers, species-specific PCR amplification and multiplex-PCR were carried out using the single species-specific primer pairs and combinations of them, respectively. Results : We used RAPD analysis of four Rheum plant species to obtain several species-specific RAPD amplicons. From nucleotide sequences of these RAPD amplicons, we developed two SCAR markers that amplified 314 bp and 390 bp DNA fragments in only R. undulatum but not in R. officinale, R. palmatum, R. tanguticum and R. undulatum, for distinguishing Rhei Undulatai Rhizoma and Rhei Radix et Rhizoma. Furthermore, we established SCAR markers for the simultaneous discrimination of the three species within a single reaction by using multiplex-PCR. Conclusions : These genetic markers can be used for the efficient discrimination of plants species and commercial herbal medicines between Rhei Undulatai Rhizoma and Rhei Radix et Rhizoma, to ultimately prevent indiscriminate distribution and prescription of these herbal medicines.