• 제목/요약/키워드: trnL

검색결과 94건 처리시간 0.022초

Discrimination of Lonicera japonica and Lonicera confusa using chemical analysis and genetic marker

  • Ryuk, Jin Ah;Lee, Hye Won;Ko, Byoung Seob
    • 대한본초학회지
    • /
    • 제27권6호
    • /
    • pp.15-21
    • /
    • 2012
  • Objective : Lonicera japonica THUNB. a traditional herbal medicine, has been commonly used anti-inflammatory disease. It has been very complicated with respect to its sources on the market. The significant selection of medicine depends on its origin. However, it is difficult to discrimination criteria for confirming L. japonica authenticity using the senses. This study was performed to determine the discriminant analysis of L. japonica and L. confusa. Methods : The identification of L. japonica and L. confusa were performed by the classification and identification committee of the national center for standardization of herbal medicines. And we examined its differences using HPLC and genetic marker analysis. Results : The analytical pattern of High Performance Liquid Chromatography was determined from the corresponding peak curves ((E)-aldosecologanin, chlorogenic acid, luteolin 7-O-glucoside, sweroside). For L. japonica, additional unknown peaks were detected at 13.8 min, 20.6 min, and 36.9 min. And, we developed genetic marker using the the tRNA-Leu gene, trnL-trnF intergenic spacer and tRNA-Phe region of chloroplast DNA. By the method, 164 bp PCR product amplified from L. confusa was distinguished into L. japonica and L. confusa efficiently. Conclusion : Base on these results, two techniques provide effective approaches to distinguish L. japonica from L. confusa.

엉겅퀴의 엽록체 TrnL-F와 Matk 영역 염기서열의 HRM 분석을 통한 특이적 SNP 분자마커의 개발 (Development of Specific SNP Molecular Marker from Thistle in the DNA Sequences of Chloroplast TrnL-F and Matk Region Using HRM Analysis)

  • 이신우;이수진;김윤희
    • 생명과학회지
    • /
    • 제29권5호
    • /
    • pp.524-529
    • /
    • 2019
  • 엉겅퀴는 대표적인 다년생의 약용식물이다. 최근 국제적 추세에 따라 자국의 유전자원의 발굴, 보존 등이 강화 됨에 따라 인접국가와 국내 자생 엉겅퀴 계통을 판별 할 수 있는 기준 설정에 관한 연구의 필요성이 대두되고 있지만, 분자생물학적 판별 기술의 개발은 아직 미흡한 실정이다. 본 연구에서는 국내 토종과 해외 유래 엉겅퀴종의 기원을 판별하기 위해 엽록체에 존재하는 trnL-trnF와 MatK 유전자단편에서 SNP를 이용한 판별 프라이머를 확보하였으며 이를 보완하여 보다 신속하게 판별하기 위하여 HRM 분석 기술을 이용한 판별 마커와 그 조건을 확립하였다. 그러므로, 본 연구에서 개발된 SNP 마커는 다양한 지역 또는 국가에서 서식하는 엉겅퀴 종들의 신속한 확인을 위해 매우 유용하게 이용될 것으로 생각된다.

Molecular Identification of Reynoutria japonica Houtt. and R. sachalinensis (F. Schmidt) Nakai Using SNP Sites

  • Park, Hana;Yoon, Chang Young;Kim, Jin Sook;Kim, Joo-Hwan
    • 한국자원식물학회지
    • /
    • 제28권6호
    • /
    • pp.743-751
    • /
    • 2015
  • Reynoutria japonica and R. sachalinensis have been used as medicinal resources in Korea. However, it is difficult to identify and determine these medicinal herbs correctly because they are usually customized and purchased as the fragmented rhizomes types. To develop molecular markers for distinguishing two species, we analyzed and compared the chloroplast DNA sequences of seven loci (atpB, matK, ccD-psaI, atpF-H, trnL-trnF, psbK-I and rpl32-trnL). Among them, we found two effective SNPs in psbK-I region for R. japonica and atpF-H region for R. sachalinensis. Based on these SNP sites, we designed the new R. japonica- specific primer which is able to amplify 300 bp fragment in psbK-I region. A similar strategy was applied for the atpF-H region of R. sachalinensis. These molecular markers would be successfully applied to recognize R. japonica and R. sachalinensis.

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
    • /
    • 제52권4호
    • /
    • pp.343-362
    • /
    • 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.

벼 색소체 형질전환을 이용한 글리포세이트 저항성 유전자 cp4-epsps의 발현 (Expression of the Glyphosate Resistant Gene, cp4-epsps, through Plastid Transformation in Rice (Oryza sativa L.))

  • 강경수;김민균
    • Journal of Plant Biotechnology
    • /
    • 제33권2호
    • /
    • pp.75-84
    • /
    • 2006
  • Heteroplasmic rice plastid transformant was generated using suspension cells as bombardment materials. PCR analyses confirmed incorporation of aadA and cp4-epsps genes into the rice plastid genome by homologous recombination events via the flanking sequences of the trnI and trnA. Transplastomic calli were actively proliferated when cultured on AAM2 medium supplemented with various concentrations (500-3000 mg/L) of streptomycin in dark condition, and transplastomic suspension cells showed resistance to nonselective herbicide, glyphosate. Through 'agarose pie selection' method, heteroplastomic calli, containing considerably high level of transplastome and expressing the CP4 EPSPS protein, were obtained. They were further regenerated to green shoots with healthy roots.

Molecular characterization of reciprocal crosses of Aerides vandarum and Vanda stangeana (Orchidaceae) at the protocorm stage

  • Kishor, Rajkumar;Devi, H.S.;Jeyaram, K.;Singh, M.R.K.
    • Plant Biotechnology Reports
    • /
    • 제2권2호
    • /
    • pp.145-152
    • /
    • 2008
  • Aerides vandarum and Vanda stangeana are two rare and endangered vandaceous orchids with immense floricultural traits. The intergeneric hybrids were synthesized by performing reciprocal crosses between them. In vitro germination response of the immature hybrid embryos was found to be best on half-strength Murashige and Skoog medium supplemented with 20% (v/v) coconut water/liquid endosperm from tender coconut. Determination of hybridity was made as early as the immature seeds or embryos germinated in vitro, using randomly amplified polymorphic DNA (RAPD) markers. Out of 15 arbitrarily chosen decamer RAPD primers, two were found to be useful in amplification of polymorphic bands specific to the parental species and their presence in the reciprocal crosses. However, a decisive profile that can identify the reciprocal crosses could not be provided by RAPD. Amplification of the trnL-F non-coding regions of chloroplast DNA of the parent species and hybrids aided easy identification of the reciprocal crosses from the fact that maternal inheritance of chloroplast DNA held true for these intergeneric hybrids. Subsequent restriction digestion of the polymerase chain reaction (PCR) amplified trnL-F non-coding regions of chloroplast DNA also consolidated the finding. Such PCR-based molecular markers could be used for early determination of hybridity and easy identification of the reciprocal crosses.

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
    • /
    • 제28권3호
    • /
    • pp.341-349
    • /
    • 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.

감국(Dendranthema indicum (L.) Des Moul.) 및 산국(D. boreale (Makino) Ling ex Kitam.)의 종판별 분자마커 개발 (Development of molecular marker for species authentication of Dendranthema indicum (L.) Des Moul. and D. boreale (Makino) Ling ex Kitam.)

  • 변지희
    • 한국자원식물학회:학술대회논문집
    • /
    • 한국자원식물학회 2018년도 추계학술대회
    • /
    • pp.66-66
    • /
    • 2018
  • 국화과(Compositae) 다년생 초본인 산국속(Dendranthema)은 국내 약 13여종이 자생하는 것으로 알려져 있으며, 이 중 감국(D. indicum (L.) Des Moul.)과 산국(D. boreale (Makino) Ling ex Kitam.), 구절초(D. zawadskii var. latilobum (Maxim.) Kitam.)가 주로 차 또는 한약재 등의 원료로 이용되고 있다. 차로 이용되는 꽃은 산국이 감국에 비해 상대적으로 작아서 구분이 가능하지만 시중에는 건조된 형태로 가공 유통되므로 육안으로 구분이 쉽지 않고, 산국 유래 제품들은 국내에서 감국 또는 국화로 혼용해서 표기되어 유통되고 있어 그 기원을 명확히 정립할 필요가 있다. 이에 본 연구는 감국과 산국의 분자유전학적 판별을 위해 DNA 바코드 후보 유전자를 활용하여 염기서열분석으로 확보된 SNP 및 InDel 정보를 바탕으로 CAPS 마커를 개발하고자 수행되었다. 감국과 산국 모두 trnL-trnF intergenic spacer 구간에서 약 1kb의 PCR 산물이 확인되었고, 이들 염기서열에서 분석한 2 SNP 및 3 InDel을 대상으로 CAPS 마커 개발을 위한 제한효소 사이트를 탐색하였다. Gap을 포함한 774bp (감국/산국=A/G) 위치의 SNP에서 BstUI(GC^GC)처리로 CAPS 마커로 전환 가능함이 확인되었고, 이에 감국과 산국의 PCR 산물에 제한효소를 처리한 결과, 제한효소 인식 사이트가 존재하는 산국에서 두 개의 DNA 단편이 확인되었다. 위 결과는 다양한 형태로 가공 유통되는 감국과 산국의 판별을 위한 마커로 활용될 수 있으며, 본 연구에 활용된 기술은 추후 건강기능식품 개발을 위한 원료표준화 확립 연구에 유용할 것으로 판단된다.

  • PDF

홍도고들빼기의 형태 다양성 및 잡종 기원의 분자 증거 (Morphological and molecular evidence of the hybrid origin of Crepidiastrum ×muratagenii in Korea)

  • 장영종;박범균;손동찬;최병희
    • 식물분류학회지
    • /
    • 제52권2호
    • /
    • pp.85-96
    • /
    • 2022
  • 홍도고들빼기는 이전 연구에서 형태적 특성과 지리적 분포를 바탕으로 이고들빼기와 갯고들빼기의 잡종인 Crepidiastrum ×muratagenii로 제안된 바 있지만, 이에 대한 분자적 증거를 제시하지 못하였다. 본 연구는 홍도고들빼기의 잡종 기원을 밝히기 위하여 홍도고들빼기와 그 근연종의 추가적인 형태적 형질을 관찰하였으며, 핵리보솜 internal transcribed spacer (ITS) 구간과 엽록체 구간(trnT-L, trnL-F, rpl16 intron, rps16 intron)의 염기서열을 비교·분석하였다. 형태적 특성을 검토한 결과, 잡종형은 생육형, 줄기잎, 외총포편, 수과의 특성을 바탕으로 세 가지 유형으로 구분되었다. 분자 분석 결과, Type 1형과 Type 2형은 ITS 구간의 종식 별부위에서 혼성화가 관찰되었으며, 엽록체 구간에서는 Type 1형은 이고들빼기, Type 2형은 갯고들빼기 서열이 각각 관찰되었다. Type 3형은 ITS와 엽록체 구간 모두 이고들빼기와 동일한 서열을 보였다. Type 1형과 Type 2형은 이고들빼기와 갯고들빼기의 형태가 혼합되어 나타날 뿐 아니라, 분자 분석에서도 절영풀이 아닌, 갯고들빼기와 이고들빼기의 종식별부위에서 혼성화가 관찰되어 이고들빼기와 갯고들빼기의 잡종임을 지지하였다. 그러나 Type 3형은 형태적 형질이 다른 잡종형과 유사하나 외총포편이 이고들빼기와 유사한 점에서 구분되며, 분자 분석에서도 이고들빼기 서열과 동일하여, 이고들빼기의 생태변이로 판단되었다.