• 제목/요약/키워드: DNA barcoding analysis

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Identification and Characterization of Microbial Community in the Coelomic Fluid of Earthworm (Aporrectodea molleri)

  • Yakkou, Lamia;Houida, Sofia;Dominguez, Jorge;Raouane, Mohammed;Amghar, Souad;Harti, Abdellatif El
    • 한국미생물·생명공학회지
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    • 제49권3호
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    • pp.391-402
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    • 2021
  • Earthworms play an important role in soil fertilization, interacting continually with microorganisms. This study aims to demonstrate the existence of beneficial microorganisms living in the earthworm's immune system, the coelomic fluid. To achieve this goal, a molecular identification technique was performed, using cytochrome c oxidase I (COI) barcoding to identify abundant endogenic earthworms inhabiting the temperate zone of Rabat, Morocco. Then, 16S rDNA and ITS sequencing techniques were adopted for bacteria and fungi, respectively. Biochemical analysis, showed the ability of bacteria to produce characteristic enzymes and utilize substrates. Qualitative screening of plant growth-promoting traits, including nitrogen fixation, phosphate and potassium solubilization, and indole acetic acid (IAA) production, was also performed. The result of mitochondrial COI barcoding allowed the identification of the earthworm species Aporrectodea molleri. Phenotypic and genotypic studies of the sixteen isolated bacteria and the two isolated fungi showed that they belong to the Pseudomonas, Aeromonas, Bacillus, Buttiauxella, Enterobacter, Pantoea, and Raoultella, and the Penicillium genera, respectively. Most of the isolated bacteria in the coelomic fluid showed the ability to produce β-glucosidase, β-glucosaminidase, Glutamyl-β-naphthylamidase, and aminopeptidase enzymes, utilizing substrates like aliphatic thiol, sorbitol, and fatty acid ester. Furthermore, three bacteria were able to fix nitrogen, solubilize phosphate and potassium, and produce IAA. This initial study demonstrated that despite the immune property of earthworms' coelomic fluid, it harbors beneficial microorganisms. Thus, the presence of resistant microorganisms in the earthworm's immune system highlights a possible selection process at the coelomic fluid level.

Molecular identification of the algal pathogen Pythium chondricola (Oomycetes) from Pyropia yezoensis (Rhodophyta) using ITS and cox1 markers

  • Lee, Soon Jeong;Hwang, Mi Sook;Park, Myoung Ae;Baek, Jae Min;Ha, Dong-Soo;Lee, Jee Eun;Lee, Sang-Rae
    • ALGAE
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    • 제30권3호
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    • pp.217-222
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    • 2015
  • Pythium species (Pythiales, Oomycetes) are well known as the algal pathogen that causes red rot disease in Pyropia / Porphyra species (Bangiales, Rhodophyta). Accurate species identification of the pathogen is important to finding a scientific solution for the disease and to clarify the host-parasite relationship. In Korea, only Pythium porphyrae has been reported from Pyropia species, with identifications based on culture and genetic analysis of the nuclear internal transcribed spacer (ITS) region. Recent fungal DNA barcoding studies have shown the low taxonomic resolution of the ITS region and suggested the mitochondrial cytochrome c oxidase subunit 1 (cox1) gene as an alternative molecular marker to identify Pythium species. In this study, we applied an analysis of both the ITS and cox1 regions to clarify the taxonomic relationships of Korean Pythium species. From the results, the two closely related Pythium species (P. chondricola and P. porphyrae) showed the same ITS sequence, while the cox1 marker successfully discriminated P. chondricola from P. porphyrae. This is the first report of the presence of P. chondricola from the infected blade of Pyropia yezoensis in Asia. This finding of the algal pathogen provides important information for identifying and determining the distribution of Pythium species. Further studies are also needed to confirm whether P. chondricola and P. porphyrae are coexisting as algal pathogens of Pyropia species in Korea.

Complete mitochondrial genome of Rotunda rotundapex Miyata & Kishida 1990 (Lepidoptera: Bombycidae), which was named as Bombyx shini Park & Sohn 2002

  • Park, Jeong Sun;Kim, Min Jee;Kim, Iksoo
    • International Journal of Industrial Entomology and Biomaterials
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    • 제44권2호
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    • pp.55-64
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    • 2022
  • Bombyx shini Park & Sohn, 2002 (Lepidoptera: Bombycidae), which was listed as an endemic species in South Korea has recently been renamed as the East Asian silk moth Rotunda rotundapex Miyata & Kishida, 1990 (Lepidoptera: Bombycidae). In this study, we sequenced the complete mitochondrial genome (mitogenome) of the R. rotundapex to announce genomic characteristics and to clarify its validity with a new name. The 15,294-bp long complete mitogenome comprises a typical set of genes [13 protein-coding genes (PCGs), 2 rRNA genes, and 22 tRNA genes] and one major noncoding, A + T-rich region, with an arrangement identical to that observed in most lepidopteran mitogenomes. The A/T content of the whole mitogenome was 79.22%; however, it varied among the regions/genes as follows: A + T-rich region, 91.62%; srRNA, 84.67%; lrRNA, 83.01%; tRNAs, 81.43%; and PCGs, 77.46%. Phylogenetic analyses of 35 species in the Bombycoidea superfamily showed the sister relationship between the families Sphingidae and Bombycidae s. str., with the higher nodal support [bootstrap support (BS) = 78%]. The Saturniidae was placed as the sister to the two families, but the nodal support for this relationship was low (BS = 53%). Current R. rotundapex was placed together with previously reported con-species with the highest nodal support, forming a separate clade from Bombyx, validating that B. shini can have a new genus name, Rotunda. However, the Korean R. rotundapex showed a substantial sequence divergence at 5.28% to that originated from an individual of type locality Taiwan in 1,459-bp of COI sequences. Considering such a high sequence divergence an additional study, which includes morphological and DNA barcoding data from further extensive distributional range maybe is needed for further robust taxonomic conclusion.

Complete Chloroplast Genome assembly and Annotation of Milk Thistle (Silybum marianum) and Phylogenetic Analysis

  • Hwajin Jung;Yedomon Ange Bovys Zoclanclounon;Jeongwoo Lee;Taeho Lee;Jeonggu Kim;Guhwang Park;Keunpyo Lee;Kwanghoon An;Jeehyoung Shim;Joonghyoun Chin;Suyoung Hong
    • 한국작물학회:학술대회논문집
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    • 한국작물학회 2022년도 추계학술대회
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    • pp.210-210
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    • 2022
  • Silybum marianum is an annual or biennial plant from the Asteraceae family. It can grow in low-nutrient soil and drought conditions, making it easy to cultivate. From the seed, a specialized plant metabolite called silymarin (flavonolignan complex) is produced and is known to alleviate the liver from hepatitis and toxins damages. To infer the phylogenetic placement of a Korean milk thistle, we conducted a chloroplast assembly and annotation following by a comparison with existing Chinese reference genome (NC_028027). The chloroplast genome structure was highly similar with an assembly size of 152,642 bp, an 153,202 bp for Korean and Chinese milk thistle respectively. Moreover, there were similarities at the gene level, coding sequence (n = 82), transfer RNA (n = 31) and ribosomal RNA (n = 4). From all coding sequences gene set, the phylogenetic tree inference placed the Korean cultivar into the milk thistle clade; corroborating the expected tree. Moreover, an investigation the tree based only on the ycf1 gene confirmed the same tree; suggesting that ycf1 gene is a potential marker for DNA barcoding and population diversity study in milk thistle genus. Overall, the provided data represents a valuable resource for population genomics and species-centered determination since several species have been reported in the Silybum genus.

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Dietary composition of two coexisting bat species, Myotis ikonnikovi and Plecotus ognevi, in the Mt. Jumbong forests, South Korea

  • Sungbae Joo;Injung An;Sun-Sook Kim
    • Journal of Ecology and Environment
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    • 제47권4호
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    • pp.168-176
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    • 2023
  • Background: Many insectivorous bats have flexible diets, and the difference in prey item consumption among species is one of the key mechanisms that allows for the avoidance of interspecies competition and promotes coexistence within a microhabitat. In Korea, of the 24 bat species that are known to be distributed, eight insectivorous bats use forest areas as both roosting and foraging sites. Here, we aimed to understand the resource partitioning and coexistence strategies between two bat species, Myotis ikonnikovi and Plecotus ognevi, cohabiting the Mt. Jumbong forests, by comparing the differences in dietary consumption based on habitat utilization. Results: Upon examining their dietary composition using the DNA meta-barcoding approach, we identified 403 prey items (amplicon sequence variants). A greater prey diversity including Lepidoptera, Diptera, Coleoptera, and Ephemeroptera, was detected from M. ikonnikovi, whereas most prey items identified from P. ognevi belonged to Lepidoptera. The diversity index of prey items was higher for M. ikonnikovi (H': 5.67, D: 0.995) than that for P. ognevi (H': 4.31, D: 0.985). Pianka's index value was 0.207, indicating little overlap in the dietary composition of these bat species. Our results suggest that M. ikonnikovi has a wider diet composition than P. ognevi. Conclusions: Based on the dietary analysis results, our results suggests the possibility of differences in foraging site preferences or microhabitat utilization between two bat species cohabiting the Mt. Jumbong. In addition, these differences may represent one of the important mechanism in reducing interspecific competition and enabling coexistence between the two bat species. We expected that our results will be valuable for understanding resource partitioning and the coexistence of bats inhabiting the Korean forests.

COI 염기서열 기반 백강잠 신속 감별용 SCAR marker 개발 - 백강잠 유전자 감별 - (Development SCAR marker for the rapid authenticaton of Batryticatus Bombyx based on COI Sequences)

  • 김욱진;양선규;노푸름;박인규;최고야;송준호;문병철
    • 대한본초학회지
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    • 제34권5호
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    • pp.13-20
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    • 2019
  • Objectives : To ensure the safety, quality and pharmacological efficacy of Batryticatus Bombyx, it is important to discriminate with adulterants. In Korean Herbal Pharmacopoeias (KHP), the authentic species of Batryticatus Bombyx is defined only Bombyx mori. Therefore, the aim of this study is establishment of PCR assay method using the sequence characterized amplified region (SCAR) marker based on COI DNA barcode for discriminating six species related to Batryticatus Bombyx. Methods : Seventeen samples of six species (Bombyx mori, Bombyx mandarina, Rhodinia fugax, Oberthueria caeca, Actias artemis, and Caligula japponica) were collected from different habitate and nucleotide sequences of cytochrome c oxidase subunit I(COI) barcode regions were analyzed by Sanger sequencing methods. To develop SCAR-based PCR assay method, we designed species-specific primers based on COI sequence variabilities and verified those specificities using 17 samples of six species as well as commercial herbal medicines. Results : In comparative multiple analysis of COI sequences, six species were distinguished by species-specific nucleotides at the species level. To develop rapid and reliable PCR assay method for genetic authentication of Batryticatus Bombyx, therefore, we designed species-specific SCAR primers based on these nucleotide sequences and confirmed those specificities. Using these SCAR primers, We also established simple conventional PCR assay method using these SCAR primers at the species level. Conclusions : The comparative analysis of COI sequences and SCAR-based PCR assay methods represented equal results for distinguishing authentic Batryticatus Bombyx and adulterations at the species level. Therefore, our results are expected protecting adulteration of herbal medicine Batryticatus Bombyx.

국내 온라인 유통 복어 제품의 종판별 및 표시사항 모니터링 연구 (Species Identification and Monitoring of Labeling Compliance for Commercial Pufferfish Products Sold in Korean On-line Markets)

  • 이지영;김건희;강태선
    • 한국식품위생안전성학회지
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    • 제38권6호
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    • pp.464-475
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    • 2023
  • 본 연구에서는 온라인 마켓에서 판매되는 50개 복어제품의 종판별 및 표시사항 일치여부 모니터링을 수행했다. 복어의 종판별을 위해 cytochrome c oxidase subunit I 및 cytochrome b 유전자의 염기서열을 분석하여 NCBI GenBank 데이터베이스에 등록되어있는 생물종의 염기서열과 비교 후 계통 분석을 수행했다. 참복, 흰점참복, 까치복, 복섬, 검복, 국매리복, 흑밀복 총 7종이 동정되었으며, 35개 제품(70%)에서 표시사항과의 불일치를 나타냈다. 12개의 제품(24%)에서는 식품공전에서 제시한 식용 가능한 복어 21종의 국명 대신 일반명(복어)을 사용하였다. 가공 정도별 불일치율은 다중가공 제품(n=9, 81.8%)이 단순가공 제품(n=26, 66.7%)보다 높은 비율을 보였으며, 원산지별 불일치율의 경우 중국산 제품(n=8, 80%)이 국산 제품(n=26, 66.7%)보다 높은 비율을 보였다. 시장명, 방언 등의 이름이 혼용되어 다수의 복어종을 밀복, 졸복으로 표시하였다. 이러한 분류체계의 어려움으로 인해 흰점참복, 국매리복과 같은 식용불가 복어가 식용 가능한 복어인 졸복으로 혼용되어 판매되는 것이 확인되었다. 따라서 식용 가능한 복어를 정확히 분류할 수 있는 방법의 개발이 필요하며, 수입 및 국내 유통 복어 제품의 주기적인 모니터링이 필요하다.

동해 근해에 출현하는 어류 자치어 분포 특성 (Distribution of Larval Fishes off the East Sea, Korea)

  • 최희찬;정해근;조정현;윤석현;오현주
    • 한국어류학회지
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    • 제34권3호
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    • pp.186-200
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    • 2022
  • 동해 근해에 출현하는 어류 자치어의 분포특성을 밝히기 위해 2018년 2월부터 12월까지 격월로 RN80 네트를 이용하여 자치어를 채집하였고, 해양 환경과 자치어 분포의 관계를 파악하기 위하여 수온, 염분 등 물리적 요인과 영양염, 엽록소-a 및 동물플랑크톤 현존량과 같은 화확 및 생물학적 요인들을 조사하였다. 분석결과 2018년 동해 근해에 출현한 자치어는 49개과 104개 분류군, 총 7,644.3개체/1,000 m3가 출현하였다. 이중 멸치가 전체 출현량의 76.2% 우점하였고, 앨퉁이가 15.0%로 그 뒤를 이었다. 그 외 까나리, 악어치가 각각 0.8%와 0.5%로 비교적 많은 출현량을 보였고, 고등어와 날붕장어가 0.3% 이상으로 위 6개 분류군이 전체 출현량의 93.2%를 차지하였다. 조사 해역의 월별 자치어 출현양상을 보면, 출현분류군수는 2월에 10개로 가장 적었고, 10월에 33개로 가장 다양한 분류군이 출현하였다. 출현량은 4월에 76.3개체/1,000 m3가 출현하여 연중 가장 적었고, 8월에 4,585.8개체/1,000 m3로 가장 많은 출현량을 보였으며, 월별로 유의한 차이를 보였다. 출현 자치어의 월별 공간 분포는 저수온기보다 고수온기에 연안 가까이에서 많이 출현하는 경향을 보였으며, 이는 출현 분류군 중 전체 출현량의 91.3%를 차지한 멸치와 앨퉁이의 월별 공간 분포와 유사하였다. 한편 자치어 전체 출현량과 해양 환경과의 상관관계는 2월에 수온 및 염분과 양의 상관관계를 보였고, 용존무기질소와 음의 상관관계를 보였다. 6월에는 용존무기질소와 음의 상관관계, 12월에는 엽록소-a와 양의 상관관계를 보였다. 최우점종인 멸치는 4월에 염분, 6월에 엽록소-a 8월에 용존무기질소와 유의한 상관관계를 보였다. ANOSIM 분석 결과, 자치어의 월별 종조성은 유의한 차이를 보였으며, 각 월별 dissimilarity는 주로 멸치와 앨퉁이의 출현량 변동에 의해 결정되었으나, 그 외 우점 분류군들의 종조성과 출현량 변화가 조사 시기별로 유의한 차이에 영향을 미쳤다. 정준대응분석 결과 자치어 군집의 분포가 저수온기에는 주로 수온, 고수온기에는 염분, 동물플랑크톤 건중량 및 영양염류 농도 등 비교적 다양한 환경요인에 영향을 받고 있었다.