• Title/Summary/Keyword: trnL

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DNA Analysis of Ginseng Using PCR-aided RFLP Technology (PCR-aided RFLP기술을 이용한 인삼의 DNA분석)

  • Yang, Deok-Chun;Kim, Moo-Sung
    • Journal of Ginseng Research
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    • v.27 no.3
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    • pp.146-150
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    • 2003
  • This study was carried out to obtain basic information on breeding using PCR-aided RFLP technology which can identify the variation inter- and intra-species of ginseng in the level of DNA. It was intended to investigate banding pattern on psbA and rbeL genes of chloroplast DNA in ginseng after treating with restriction enzymes. To isolate psbA and rbcL genes of chloroplast, both psbA-N, psbA-C primer and rbcL-N, PX-1 primer were used. As a result, 1,008 bp band of psbA gene and 1,336 bp band of rbcL gene were appeared, which was optimal and expected molecular weight. In addition, primers to isolate atpB, rpoB, trnL, and trnF genes were used, resulting in the expected 1366, 900, 1500 and 1008 bp bands. Genes of psbA and rbcL isolated by PCR were cut by restriction enzymes, Sau3A, TaqI, AluI, HaeIII, and RFLP pattern was investigated. KG line and other species of ginseng were cut by TaqI treatment, and bands were located in 800 bp. The treatment treated by AluI also showed the same 800 bp band in KG line and other species. In HaeIII treatment, 500 bp of faint bands were shown in case of KG line, whereas any bands were not observed in other species. All chloroplast genes formed bands by PCR amplification. However, it was not evident to distinguish intra-or inter-species of ginseng after restriction enzyme treatment. Therefore, more restriction enzyme treatment or sequence comparison method should be considered for further experiment.

Improved plastid transformation efficiency in Scoparia dulcis L.

  • Kota, Srinivas;Hao, Qiang;Narra, Muralikrishna;Anumula, Vaishnavi;Rao, A.V;Hu, Zanmin;Abbagani, Sadanandam
    • Journal of Plant Biotechnology
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    • v.46 no.4
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    • pp.323-330
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    • 2019
  • The high expression level of industrial and metabolically important proteins in plants can be achieved by plastid transformation. The CaIA vector, a Capsicum-specific vector harboring aadA (spectinomycin resistance), is a selectable marker controlled by the PsbA promoter, and the terminator is flanked by the trnA and trnI regions of the inverted repeat (IR) region of the plastid. The CaIA vector can introduce foreign genes into the IR region of the plastid genome. The biolistic method was used for chloroplast transformation in Scoparia dulcis with leaf explants followed by antibiotic selection on regeneration medium. Transplastomes were successfully screened, and the transformation efficiency of 3 transgenic lines from 25 bombarded leaf explants was determined. Transplastomic lines were evaluated by PCR and Southern blotting for the confirmation of aadA insertion and its integration into the chloroplast genome. Seeds collected from transplastomes were analyzed on spectinomycin medium with wild types to determine genetic stability. The increased chloroplast transformation efficiency (3 transplastomic lines from 25 bombarded explants) would be useful for expressing therapeutically and industrially important genes in Scoparia dulcis L.

Molecular Phylogenetic Studies of Korean Hydrocotyle L. (한국산 피막이속(Hydrocotyle L.) 식물의 분자계통학적 연구)

  • Choi, Kyoung-Su;Park, Seon-Joo
    • Korean Journal of Plant Resources
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    • v.25 no.4
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    • pp.490-497
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    • 2012
  • Phylogenetic analyses were conducted to evaluate relationships of 5 taxa of Korean Hydrocotyle, H. spp. found in the Ulleung island including one outgroup (Centella asiatica). The molecular phylogenetic methods based on nuclear ribosomal DNA ITS region and cpDNA trnH-psbA region. Centella asiatica was used outgroup for analysis. As the result, genus Korean Hydrocotyle were grouped by 94% bootstrap value. Korean Hydrocotyle was grouped by four clades; Clade I-H. maritima, H. sibthorpides and H. yabei clade Clade II-H. nepalensis clade clade III-H. ramiflora clade clade IV-H. spp. clade. H. maritima, H. sibthorpides and H. yabei was not distinguished, seperately. H. spp. was distinctly distinguished other Korean Hydrocotyle.

Taxonomic Review of the Genus Echinochloa in Korea (I): Inferred from Sequences of cpDNA and nrDNA

  • Lee, Jeongran;Kim, Chang-Seok;Lee, In-Yong
    • Weed & Turfgrass Science
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    • v.3 no.3
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    • pp.183-189
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    • 2014
  • The genus Echinochloa (L.) P. Beauv. comprised of approximately 30-40 species in the tropical and warm temperate regions of the world, including numerous interspecific and intraspecific types which make the genus difficult to identify. As an attempt to identify the species within the genus easier, the taxonomy of the genus Echinochloa, Poaceae in Korea was reviewed on the basis of sequencing data derived from nuclear ribosomal DNA internal transcribed spacer (ITS) and external transcribe spacer and chloroplast DNA trnL intron, trnL-F intergenic spacer and matK regions using a total of 46 accessions representing all the species in Korea. The results of maximum parsimony found separate lineage comprised of E. colona and E. frumentaceae which are not Korean species, but no resolution within Korean Echinochloa species, supporting the suggestion of Yamaguchi group that E. crus-galli, E. oryzoides, and E. esculenta should be considered to belong to the same species. However, the relationship between these three species and the other species, i.e. E. oryzicola should be better understood with more detail studies.

Application for Identification of Food Raw Materials by PCR using Universal Primer (일반 프라이머를 이용한 PCR의 식품원료 진위 판별에 적용)

  • Park, Yong-Chjun;Jin, Sang-Ook;Lim, Ji-Young;Kim, Kyu-Heon;Lee, Jae-Hwang;Cho, Tae-Yong;Lee, Hwa-Jung;Han, Sang-Bae;Lee, Sang-Jae;Lee, Kwang-Ho;Yoon, Hae-Seong
    • Journal of Food Hygiene and Safety
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    • v.27 no.3
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    • pp.317-324
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    • 2012
  • In order to determine an authenticity of food ingredient, we used DNA barcode method by universal primers. For identification of animal food ingredients, LCO1490/HCO2198 and VF2/FISH R2 designed for amplifying cytochrome c oxidase subunit1 (CO1) region and L14724/H15915 for cytochrome b (cyt b) region on mitochondrial DNA were used. Livestock (cow, pig, goat, sheep, a horse and deer) was amplified by LCO1490/HCO 2198, VF2/FISH R2 and L14724/H15915 primers. Poultry (chicken, duck, turkey and ostrich) was amplified by LCO1490/HCO 2198 and VF2/FISH R2 primers. But, Fishes (walleye pollack, herring, codfish, blue codfish, trout, tuna and rockfish) were only amplified by VF2/FISH R2 primers. For plant food ingredients, 3 types of primers (trnH/psbA, rpoB 1F/4R and rbcL 1F/724R) have been used an intergenic spacer, a RNA polymerase beta subunit and a ribulose bisphosphate carboxylase region on plastid, respectively. Garlic, onion, radish, green tea and spinach were amplified by trnH/psbA, rpoB 1F/4R and rbcL 1F/724R. The PCR product sizes were same by rpoB 1F/4R and rbcL 1F/724R but, the PCR product size using trnH/psbA primer was different with others for plants each. We established PCR condition and universal primer selection for 17 item's raw materials for foods and determine base sequences aim to PCR products in this study. This study can apply to determine an authenticity of foods through making an comparison between databases and base sequences in gene bank. Therefore, DNA barcode method using universal primers can be a useful for species identification techniques not only raw materials but also processed foods that are difficult to analyze by chemical analysis.

DNA Yield and PCR Success Rate of the Establishment Time of Wood Annual Ring: A Case Study of Korean Red Pine (Pinus densiflora) (목재의 나이테 생성 시기에 따른 DNA 추출 수율 및 PCR 성공률: 소나무(Pinus densiflora) 목재의 사례)

  • So Hyeon Kim;Byeong-Ju Lee;Ji Young Ahn;Jei-Wan Lee;Hyun-Mi Lee;Soo Hyung Eo
    • Journal of Korean Society of Forest Science
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    • v.112 no.4
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    • pp.554-560
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    • 2023
  • To prevent illegal timber distribution, DNA markers have been used to identify the species and origin. However, extracting high-quality DNA from timber is difficult because of its physical and chemical properties. In this study, we investigated whether the age of timber tissue influences the yield of DNA extraction and the success rate of polymerase chain reaction (PCR) to understand the relationship between the establishment time of the wood annual ring and the extracted DNA concentration (ng/μl), purity (A260/A280), and PCR success rate (%) from pinewood, a major Korean domestic species. According to the results, it was observed that as the distance from the cambium increased, indicating that the tissue was older, the concentration and purity of the extracted DNA decreased significantly. For the trnM-trnV (285 bp) and rpoC1 (298 bp) regions, the PCR success rate was 100%. However, for the rbcL (1.3 kb) region, the PCR success rate was 66.67%. Moreover, PCR amplification of the rbcL region failed at all points older than 30 years. Thus, it is deduced that as time passes, along with the decay of timber cells, DNA is degraded, leading to a decrease in DNA concentration, purity, and PCR success rate. The results of this study are expected to be beneficial for future applications, such as the species identification of timber, providing valuable insights and potential utilization in this field.

Taxonomic Review on Hydrangea macrophylla (Thunb.) Ser. and H. serrata f. acuminata (Siebold & Zucc.) E.H.Wilson (산수국과 수국의 분류학적 재검토)

  • Kang, Shin-Ho;Chung, Kyong-Sook
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2019.10a
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    • pp.22-22
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    • 2019
  • 수국속(Hydrangea L.)은 APG IV분류체계에 따르면 국화군(Asterids), 층층나무목(Cornales), 수국과(Hydrangeaceae)에 속하는 식물로서 동아시아, 북아메리카 동부 지역을 중심으로 분포하며 전 세계에 23여종이 알려져 있다. 한국산 수국속은 Nakai가 H. hortensia var. acuminata의 분포를 기록함으로써 시작되었다. 이후 Nakai는 산수국을 H. acuminata로 재처리, 다시 H. serrata var. acuminata로 변경하였다. 한편 산수국은 분류학적 위치에 대한 이견이 있다. Wilson은 수국과 산수국을 가까운 유연관계를 가지는 독립된 분류군으로 분류하였지만, Makino는 산수국을 수국의 아종인 H. marcrophylla subsp. serrta로 기재하는 등 산수국과 수국의 동정과 계급설정에 혼란이 있는 가운데, 최근 천연물 hydrangenol을 활용한 건강기능식품 개발에서 사용가능 재료의 한계를 설정하기 위한 산수국과 수국에 대한 종 설정 연구도 요구되고 있다. 따라서 본 연구에서는 nrDNA ITS, trnL-F, trnC-ycf6의 3개 구간의 DNA sequence와 NCBI (National Center for Biotechnology Information)를 참고한 분자생물학적 분석결과와 외부형태학적 재검토 결과를 통하여 H. marcrophylla를 종(Species)으로서의 분류학적 위치 지지하며 또한 수국과 산수국을 가까운 유연관계를 가지는 독립된 분류군으로 분류한 의견을 지지하는 결과를 도출하였다. 향후 hydrangenol 활용의 기초정보로 활용되기를 기대한다.

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Identification of three independent fern gametophytes and Hymenophyllum wrightii f. serratum from Korea based on molecular data

  • LEE, Chang Shook;LEE, Kanghyup;HWANG, Youngsim
    • Korean Journal of Plant Taxonomy
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    • v.50 no.4
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    • pp.403-412
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    • 2020
  • Colonies of three independent gametophytes (one that is filamentous and two that are ribbon-like) without sporophytes occur in Gyeonggi-do, Gangwon-do, Gyeongsang-do, and Jeju-do, Korea. They have a moss-like appearance at first sight, with tiny plantlets and gemmae, and grow in cool, shaded, relatively deep dint places of large rocks, such as the small caves in high mountains, close to valleys. The gametophytes were identified based on morphological and molecular data by chloroplast DNA (cpDNA) sequence data (rbcL, rps4 gene and rps4-trnS intergenic spacer). Here, rbcL, rps4 gene and rps4-trnS intergenic spacer data of one independent gametophyte distributed in Korea have the same morphology, DNA sequence and monophyletic group as Crepidomanes intricatum from the eastern United States. They also share the same cpDNA data with Crepidomanes schmidtianum recently reported from Korea. The other independent gametophyte should be Hymenophyllum wrightii based on cpDNA data. The last one was presumed to be Pleurosoriopsis makinoi based on molecular data. The taxonomic status was confirmed to be the forma of Hymenophyllum wrightii through a revision of Hymenophyllum wrightii f. serratum based on molecular data.

Phylogenetic relationships of Korean campanulaceae based on chloroplast DNA sequences (엽록체 DNA 염기서열 분석을 이용한 한국산 초롱꽃과 (Campanulaceae)의 계통유연관계)

  • Kim, Kyung-Ah;Yoo, Ki-Oug
    • Korean Journal of Plant Taxonomy
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    • v.42 no.4
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    • pp.282-293
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    • 2012
  • Phylogenetic studies were conducted to evaluate the taxonomic relationships among 28 taxa, including 2 outgroups of Korean Campanulaceae, using atpB, atpB-rbcL, atpF-H, matK, rbcL, rpl16, rpoC1 and trnL-F regions sequences in chloroplast DNA. The combined analyses of eight chloroplast DNA regions suggest that Codonopsis and Platycodon basally branches within the phylogenetic tree; Wahlenbergia distinguished an independent clade; Campanula forms a clade; Peracarpa and Asyneuma clade is a sister to the Adenophora-Hanabusaya clade; Hanabusaya is placed within the section Remotiflorae of Adenophora; Adenophora form a clade. Our present results support the generic level, although discordance remained at the infrageneric groups such as section and series based on morphological characteristics in the genus Adenophora.

The Study of DNA markers to identify of Allium sativum L. (한약재 마늘(Allium sativum L.)의 식별을 위한 유전자 감식연구)

  • Son, OGyeong;Seo, Bu-II;Lee, Seon-Ha;Park, Seon-Joo
    • The Korea Journal of Herbology
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    • v.29 no.1
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    • pp.27-33
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    • 2014
  • Objectives : This study was carried out to identify DNA markers of "Allium sativum" be circulated from Korea and China, which is difficult to discriminate from morphological characters because of fragmental materials of bulb. That is, all these studies focused on the discrimination of Allium sativum L. But these day, Chinese A. sativum was in circulated Korean A. sativum in Korean medicine markets. Therefore, the purpose of our study was to develop molecular markers for discrimination between Korean A. sativum and imports from China. Methods : Materials were collected randomly from a markets in Korea and China and be analyzed with matK, ndhF and trnL-F regions of chloroplast DNA (cpDNA). We collected 45 A. sativum individuals from Korean and Chinese medicine markets, in 2013. Results : As a results, matK and ndhF regions of cpDNA was shown to be identify, Species that grow from warm place and cold place can divide as five SNP (Single nucleotide polymorphisms) markers in matK and ndhF genes. Also, in trnL-F regions, found one SNP that can divide Korean A. sativum and Chinese A. sativum. Conclusions : From the analysis of matK and ndhF regions of cpDNA, we presumed that three markers of cpDNA were found by useful marker that can distinguish Korean, Chinese, Warm place type, and Cold place type. Individual differences of Korean and Chinese was thought that appear in geographical difference and genetic difference by environment for long hour even if same species.