• Title/Summary/Keyword: DNA-based Identification

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Non-Invasive Sex Determination of Asiatic Black Bear (Ursus thibetanus) via Sex-Specific Amplification of the Amelogenin Gene

  • Baek-Jun Kim
    • Proceedings of the National Institute of Ecology of the Republic of Korea
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    • v.4 no.4
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    • pp.154-158
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    • 2023
  • The Asiatic black bear, Ursus thibetanus, is among the most threatened or endangered species in Asia. For its conservation and management, sex identification of U. thibetanus using non-invasive samples (e.g., hair and/or feces) is potentially valuable. In this study, a non-invasive molecular method for sex identification of U. thibetanus samples collected from various countries was first utilized, and it was based on polymerase chain reaction (PCR) amplification of the amelogenin gene via PCRs. Thirty-three bear DNA samples, extracted not only from blood (n=9) but also from hair (n=18) and feces (n=6), were used. We performed sex-specific PCR amplifications of the amelogenin gene using a primer set, SE47 and SE48. The primer set could successfully amplify a single X-specific band for females and both X- and Y-specific bands for males from all blood (100%) and hair (100%) samples. In addition, the primer set could distinguish the sex of bears in four out of a total of six fecal samples (approximately 67%). This study's findings suggest that this molecular method can be applied to sex identification of Asiatic black bears from various Asian regions using non-invasive samples, such as hair and feces.

Molecular Authentication of Schisandrae Fructus and Analysis of Phylogenetic Relationship based on nrDNA-ITS sequences (nrDNA-ITS 분자마커를 이용한 오미자(五味子) 종 감별 및 기원분석 -ITS 염기서열을 이용한 오미자(五味子) 감별-)

  • Moon, Byeong-Cheol;Ji, Yun-Ui;Seo, Hyeong-Seok;Lee, A-Young;Chun, Jin-Mi;Kim, Ho-Kyoung
    • The Korea Journal of Herbology
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    • v.25 no.4
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    • pp.47-54
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    • 2010
  • Objectives : The original plant species of Schisandrae Fructus (O-mi-ja) is prescribed as Schisandra chinensis $B_{AILL.}$, in Korea, but S. chinensis $B_{AILL.}$ and S. sphenanthera $R_{EHD.}$ et $W_{ILS.}$ in China. Moreover, fruit of several other species in genus Schisandra also have been used as the same herbal medicines. To develop a reliable method for correct identification of Schisandrae Fructus and to evaluate the phylogenetic relationship of S. chinensis and its related species, we analyzed internal transcribed spacer (ITS) sequences of nuclear ribosomal DNA (nrDNA). Methods : Twenty-four plant samples of three Schisandra species and one Kadsura species, S. chinensis $B_{AILL.}$, S. spenanthera $R_{EHD.}$ et $W_{ILS.}$, S. nigra $M_{ax.}$ and Kadsura japonica $D_{UNAL}$ were collected from each different native habitate and farm in Korea and China. The nrDNA-ITS region of each samples were amplified using ITS1 and ITS4 primer and nucleotide sequences were determined after sub-cloning into the pGEM-Teasy vector. Authentic marker nucleotides were estimated by the analysis of ClastalW based on the entire nrDNA-ITS sequence. Results : In comparative analysis of the nrDNA-ITS sequences, we found specific nucleotide sequences including indels (insertions and deletions) and substitutions to distinguish C. chinensis, S. spenanthera, S. nigra, and K. japonica. These sequence differences at corresponding positions are avaliable nucleotide markers to determine the botanical origin of O-mi-ja. Moreover, we evaluated the phylogenetic relationship of four plant species by the analysis of nrDNA-ITS sequences. Conclusions : These marker nucleotides would be useful to identify the official herbal medicines by the providing of definitive information that can identify each plant species and distinguish it from unauthentic adulterants for O-mi-ja.

Identification of Phellinus linteus by Morphological Characteristics and Molecular Analysis (형태적.분자생물학적 방법에 의한 Phellinus linteus의 동정에 관한 연구)

  • Kim, Sang-Hee;Kim, Soo-Ho;Sung, Jae-Mo;Harrington, Thomas C.
    • The Korean Journal of Mycology
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    • v.27 no.5 s.92
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    • pp.337-340
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    • 1999
  • The context and upper surface of Phellinus basidiocarp become blackened, rimose and woody. The basidiocarp is sessile, dimidiate and elongate. The basidiospores are pigmented and ovoid to globose. Hymenial setae are $17{\sim}35{\times}6{\sim}8{\mu}m$. Nineteen isolates of Phellinus species, including Phellinus linteus, were used for sequencing of the internal transcribed spacer (ITS) region of the nuclear rDNA. Based on these sequence data, specific primers were designed for identification of Phellinus linteus isolates in Korea. The specific primers were within the ITS1 and ITS2 regions and were nested within the universal primers flanking the spacer regions. A total of four primers (the universal primers ITS-1F and ITS-4, and the specific primers PL-F and PL-R) were used for detection of Phellinus linteus collected in Korea. The length of the four amplification products of Phellinus linteus DNA were 800 bp (ITS-1F/ITS-4), two bands of about 720 bp (ITS-1F/PL-R and PL-F/ITS-4), and 610 bp (PL-F/PL-R). Among 23 isolates of Phellinus species collected in Korea, Thirteen isolates were identified as Phellinus linteus based on the presence of the four bands. The other species produced only the single ITS-1F/ITS-4 product.

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Bird accidents in Southern Mongolia: a case study of bird electrocution (몽골 남부지역의 야생조류 사고: 감전사를 중심으로)

  • Ganbold, Onolragchaa;Bing, Gi-Chang;Purevee, Erdenetushig;Munkhbayar, Munkhbaatar;Choi, Won-Suk;Jargalsaikhan, Ariunbold;Paik, In-Hwan;Purevdorj, Zoljargal;Jargal, Namsrai;Paek, Woon-Kee
    • Korean Journal of Ornithology
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    • v.25 no.2
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    • pp.94-100
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    • 2018
  • Bird electrocution on contact with electricity lines is well reported and is quite common in Mongolian open areas. We visited 15-kV electricity distribution pole lines in a Southern Mongolian semi-desert region three times in 2017, in April, July, and September, to assess their risks to birds. The carcasses of 45 electrocuted birds representing 12 species were identified from 250 poles (overall mortality rate of 1.12% every 10 km). The majority of these electrocuted birds were endangered Saker Falcon (n=11) and common Black Kite (n=11). The lacks of roosting or perching sites in our study sites (indeed other open areas in Mongolia) may resulted for such high rate bird electrocution. A 751-base pair (bp) cytochrome c oxidase I (COI) gene was amplified for DNA sequence-based identification of carcasses that were difficult to identify directly. Our findings revealed the high electrocution risk for birds in Mongolian open areas, especially for the birds of prey, with relatively low-efficiency electrocution mitigation approaches. The findings also indicate that there is a need for better understanding of the risk of bird electrocution, particularly in the open areas. This will contribute to the conservation of endangered species.

Identification of Orchid Mycorrhizal Fungi Isolated from Epipactis thunbergii in Korea (닭의난초(Epipactis thunbergii)에 공생하는 난 균근균의 분리 및 동정)

  • Han, Han-Kyeol;Chung, Jae-Min;Cho, Yong-Chan;Kim, Dae-Shin;Eom, Ahn-Heum
    • The Korean Journal of Mycology
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    • v.41 no.1
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    • pp.9-13
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    • 2013
  • In this study, roots of Epipactis thunbergii were collected from Chujado on the north of Jeju-do. Six fungal isolates were isolated from surface-sterilized roots of the orchid and classified with groups based on morphological characteristics. Fungal DNA was extracted from each isolate and amplified ITS region using ITS1-OF/ITS4-OF primer pair. Three species of orchid mycorrhizal fungi were identified as Tulasnella calospora, Tulasnella sp. and Sebacina sp. based on molecular and morphological characteristics.

PCR-Based Assay for Rapid and Specific Detection of the New Xanthomonas oryzae pv. oryzae K3a Race Using an AFLP-Derived Marker

  • Song, Eun-Sung;Kim, Song-Yi;Noh, Tae-Hwan;Cho, Heejung;Chae, Soo-Cheon;Lee, Byoung-Moo
    • Journal of Microbiology and Biotechnology
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    • v.24 no.6
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    • pp.732-739
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    • 2014
  • We describe the development of a polymerase chain reaction method for the rapid, precise, and specific detection of the Xanthomonas oryzae pv. oryzae (Xoo) K3a race, the bacterial blight pathogen of rice. The specific primer set was designed to amplify a genomic locus derived from an amplified fragment length polymorphism specific for the K3a race. The 1,024 bp amplicon was generated from the DNA of 13 isolates of Xoo K3a races out of 119 isolates of other races, pathovars, and Xanthomonas species. The assay does not require isolated bacterial cells or DNA extraction. Moreover, the pathogen was quickly detected in rice leaf 2 days after inoculation with bacteria and at a distance of 8 cm from the rice leaf 5 days later. The results suggest that this PCR-based assay will be a useful and powerful tool for the detection and identification of the Xoo K3a race in rice plants as well as for early diagnosis of infection in paddy fields.

Unrecorded Fungal Species Isolated from Indoor Air in the Log Bed- and Sawdust Media-based Mushroom Cultivation Houses (원목 및 톱밥배지 버섯 재배사 내 실내 공기서 분리한 미기록 진균 보고)

  • Ahn, Geum Ran;Kim, Ji Eun;Kim, Jun Young;Kim, Seong Hwan
    • The Korean Journal of Mycology
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    • v.46 no.4
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    • pp.495-503
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    • 2018
  • Oak mushroom is cultivated using logs and sawdust media as substrates. In this study, fungi were isolated during a monitoring of indoor air in the oak mushroom cultivation houses located in Cheongyang-gun of Chungnam, Geoje-gun of Gyeongnam, Gumi-si of Gyeongbuk, Jangheung-gun of Jeonnam and Yeoju-si of Gyeongggi-do. Identification of the fungi based on morphology and molecular analysis of the internal transcribed spacer region and 28S rDNA, translation elongation factor translation elongation factor 1 a gene, and ${\beta}-tubulin$ gene revealed that six fungi, Cenangium acuum, Neopestalotiopsis surinamensis, Metarhizium marquandii, Periconia macrospinosa, Trichoderma petersenii, and Trichoderma paratroviride that have not been recorded previously in Korea.

Mitochondrial sequence based characterization and morphometric assessment of Diara buffalo population

  • Singh, Karan Veer;Purohit, Hitesh;Singh, Ramesh Kumar
    • Animal Bioscience
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    • v.35 no.7
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    • pp.949-954
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    • 2022
  • Objective: The present study is aimed at phenotypic characterization and mitochondrial d-loop analysis of indigenous "Diara" buffalo population, which are mostly confined to the villages on the South and North Gangetic marshy plains in the Bihar state of India. These buffaloes are well adapted and are best suited for ploughing and puddling the wet fields meant for paddy cultivation. Methods: Biometric data on 172 buffaloes were collected using a standard flexible tape measure. Animals are medium in size; the typical morphometric features are long head with a broad forehead and moderately long and erect ears. Genomic DNA was isolated from unrelated animals. The mtDNA d-loop 358-bp sequence data was generated and compared with 338 sequences belonging to riverine and swamp buffaloes. Results: Based on the mitochondrial d-loop analysis the Diara buffaloes were grouped along with the haplotypes reported for riverine buffalo. Sequence analysis revealed the presence of 7 mitochondrial D loop haplotypes with haplotype diversity of 0.9643. Five of the haplotypes were shared with established swamp breeds and with Buffalo population of Orissa in India. Conclusion: Morphometric analyses clearly shows distinguishing features like long and broad forehead which may be useful in identification. The germplasm of Diara buffalo is much adapted to the marshy banks of river Ganga and its tributaries. It constitutes a valuable genetic resource which needs to be conserved on priority basis.

Species Identification and Molecular Phylogenetic Position of Korean Damselfishes (Pomacentridae: Chrominae) Based on DNA Bioinformation (DNA 생물정보를 이용한 한국산 자리돔과 어류의 분류 및 분자계통학적 위치)

  • Koh, Jeong Rack;Park, Yung Chul
    • Korean Journal of Ichthyology
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    • v.19 no.4
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    • pp.274-285
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    • 2007
  • The subfamily Chrominae of damselfishes (Teleostei: Pomacentridae) includes the genus Chromis and Dascyllus. They are found throughout the tropical oceans and form a major component of coral reef communities. There are 5 species of the Chrominae currently recognized in Korea. This study was conducted to infer phylogenetic position of two Korean Chromis species and one Dascyllus species within general category of their each genus in worldwide level. This study also includes one species of Japanese Dascyllus. In the phylogenetic analysis, the Japanese D. aruanus grouped with D. aruanus previously reported from French Polynesia. Korean Chromis fumea grouped with Australian C. nitida and the p-distance value between the two species is relatively very low (0.047). Korean C. notatus grouped together with C. flavomaculata (New Caledonia). In the sequence analysis of some Korean and Japanese damselfishes, there was no sequence variation between D. melanurus (Jeju, Korea) and D. melanurus (Indo-Pacific), but the sequences of the two populations were different in only one nucleotide sites from that of D. melanurus in Indonesian Archipelago. The sequences of Dascyllus aruanus (Japan) were different in two nucleotide sites from it in French Polynesia. There were high difference between the sequences of two Korean species, Chromis fumea and Korean C. notatus. The variations among mitochondrial cytochrome b sequences indicate that the gene sequence could be used as DNA barcode for identification of local populations of D. aruaus and D. melanurus as well as species level.

Identification and Biochemical Characterization of a New Xylan-degrading Streptomyces atrovirens Subspecies WJ-2 Isolated from Soil of Jeju Island in Korea (제주도 토양으로부터 자일란 분해 Streptomyces atrovirens subspecies WJ-2 동정 및 효소의 생화학적 특성 규명)

  • Kim, Da Som;Bae, Chang Hwan;Yeo, Joo Hong;Chi, Won-Jae
    • Microbiology and Biotechnology Letters
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    • v.44 no.4
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    • pp.512-521
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    • 2016
  • A bacterial strain was isolated from a soil sample collected on Jeju Island, Korea. The strain, designated WJ-2, exhibited a high xylanase activity, whereas cellulase activity was not detected. The 16S rRNA gene sequence of WJ-2 was highly similar to type strains of the genus Streptomyces. A neighbor-joining phylogenetic tree based on 16S rRNA gene sequences showed that strain WJ-2 is phylogenetically related to Streptomyces atrovirens. Furthermore, DNA-DNA hybridization analysis confirmed that strain WJ-2 is a novel subspecies of Streptomyces atrovirens. The genomic DNA G+C content was 73.98 mol% and the major fatty acid present was anteiso-C15:0 (36.19%). The growth and xylanase production of strain WJ-2 were significantly enhanced by using soytone and xylan as nitrogen and carbon sources, respectively. Crude enzyme preparations from the culture broth of strain WJ-2 exhibited maximal total xylanase activities at pH 7.0 and $55^{\circ}C$. Thin-layer chromatography analysis revealed that the crude enzyme degrades beechwood xylan to yield xylobiose and xylotriose as the principal hydrolyzed end products.