• Title/Summary/Keyword: trnL

Search Result 94, Processing Time 0.025 seconds

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

  • Ryuk, Jin Ah;Lee, Hye Won;Ko, Byoung Seob
    • The Korea Journal of Herbology
    • /
    • v.27 no.6
    • /
    • 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.

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

  • Lee, Shin-Woo;Lee, Soo Jin;Kim, Yun-Hee
    • Journal of Life Science
    • /
    • v.29 no.5
    • /
    • pp.524-529
    • /
    • 2019
  • Medicinal plants resources are becoming important assets since their usages have been expanded to the development of functional foods for human health, cosmetics and pharmaceutical industries. However, their phylogenetic origins and names are different from each country and quite often they are mixed each other resulting in the confusion for consumers. Particularly when they are very similar based on their morphological characteristics and distributed, it is extremely difficult to differentiate their origins even by specialists. Therefore, identification of each plant species is important for standardizing herbal medicine. Thistle is a medicinal and perennial plant. Obtaining information about the genetic diversity of plant populations is highly important for conservation and germplasm utilization. Although thistle is an important medicinal plant species registered in South Korea, no molecular markers are currently available to distinguish from other similar species from different countries. In this study, we developed single nucleotide polymorphism (SNP) markers derived from chloroplast genomic sequences to identify distinct Korean-specific thistle species via high resolution melting (HRM) curve analyses. We performed molecular authentication of four different kinds of thistle species from different regions using DNA sequences in the trnL-F and matK chloroplast intergenic region. The SNP markers developed in this study are useful for rapidly identifying specific thistle species from different country.

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
    • Korean Journal of Plant Resources
    • /
    • v.28 no.6
    • /
    • 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
    • /
    • v.52 no.4
    • /
    • 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.

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

  • Kang, Kyung-Su;Kim, Min-Kyun
    • Journal of Plant Biotechnology
    • /
    • v.33 no.2
    • /
    • 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
    • /
    • v.2 no.2
    • /
    • 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
    • Korean Journal of Plant Resources
    • /
    • v.28 no.3
    • /
    • 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.

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

  • Byeon, Jihui
    • Proceedings of the Plant Resources Society of Korea Conference
    • /
    • 2018.10a
    • /
    • 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

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

  • JANG, Young-Jong;PARK, Boem Kyun;SON, Dong Chan;CHOI, Byoung-Hee
    • Korean Journal of Plant Taxonomy
    • /
    • v.52 no.2
    • /
    • pp.85-96
    • /
    • 2022
  • The plant "Hong-do-go-deul-ppae-gi" has been considered as Crepidiastrum × muratagenii, a hybrid between C. denticulatum and C. lanceolatum, based on its morphological traits and geographical distribution. To reveal the hybrid origin of Hong-do-go-deul-ppae-gi, we examined additional morphological traits of this plant and its putative parents (C. denticulatum, C. lanceolatum, C. platyphyllum) and analyzed one nuclear ribosomal internal transcribed spacer (ITS) region and four chloroplast regions (trnT-L, trnL-F, rpl16 intron, and rps16 intron). As a result of examining the morphological traits, putative hybrid individuals were classified into three types based on the habit, cauline leaf, outer phyllary, and achene beak traits. A molecular analysis found that the ITS sequences of Type 1 and Type 2 individuals showed additive species-specific sites of C. denticulatum and C. lanceolatum. Plastid sequences of Type 1 and Type 2 individuals showed C. denticulatum and C. lanceolatum sequences, respectively. However, Type 3 individuals had ITS and plastid sequences corresponding to C. denticulatum. Accordingly, Type 1 and Type 2 individuals not only share morphological traits with C. denticulatum and C. lanceolatum but also show additive species-specific sites for C. denticulatum and C. lanceolatum, and not C. platyphyllum, supporting its origin as a hybrid between C. denticulatum and C. lanceolatum. Type 3 had morphological traits similar to other hybrid types but was distinguished with respect to outer phyllaries and demonstrated some resemblance to C. denticulatum. In a molecular analysis, Type 3 was found to be identical with regard to the sequence of C. denticulatum and was judged to be an ecological variation of C. denticulatum.