• Title/Summary/Keyword: SPECIES IDENTIFICATION

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Molecular Identification of Thrips in Two Medicinal Crops, Cnidium officinale Makino and Ligusticum chuanxiong Hort (일천궁과 토천궁에서 발생하는 총채벌레류의 분자동정)

  • Jung, Chung Ryul;Jeong, Dae Hui;Park, Hong Woo;Kim, Hyun Jun;Jeon, Kwon Seok;Yoon, Jung Beom
    • Korean Journal of Medicinal Crop Science
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    • v.27 no.1
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    • pp.17-23
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    • 2019
  • Background: Cnidium officinale Makino and Ligusticum chuanxiong Hort. are important medicinal crops in Korea. However, there is insufficient information on the identification of thrips, which attack these plants. Until now, one species of thrips has been recorded as a main pest. Methods and Results: To identify the thrips emerging in C. officinale Makino and L. chuanxiong Hort., these plants were independently cultivated in two local areas. Thirty individuals of each plant species were selected randomly and surveyed for the presence of thrips. After confirming the existence of thrips, 100 thrips individuals were collected from each crop using the beating method. To identify thrips species, we performed PCR-restriction fragment length polymorphism (RFLP)-based analysis using ITS2 primer sets. Six thrips species were identified: western flower thrips (Frankliniella occidentalis), flower thrips (F. intonsa), onion thrips (Thrips tabaci), chrysanthemum thrips (T. nigropilosus), chilli thrips (Scirtothrips dorsalis), and grass thrips (Anaphothrips obscurus). The proportion of these species differed between the host plant species. Conclusions: Six thrips species were major pests of two medicinal crops. Integrated pest management is required to control these thrips species, and will enhance the yield and quality of C. officinale and L. chuanxiong.

Animal species identification by co-amplification of hypervariable region 1 (HV1) and cytochrome b in mitochondrial DNA

  • Lim, Si Keun;Park, Ki Won
    • Analytical Science and Technology
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    • v.18 no.3
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    • pp.257-262
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    • 2005
  • Mitochondrial DNA (mt DNA) sequence analysis has been a useful tool for species identification of animals and human individuals. Two hypervariable regions (HV1 and HV2) in control region of mitochondrial genome were analyzed for human individual identification. In case of animal species identification, several genes on mt DNA such as cytochrome b (cytb), RNAs, cytochrome oxidases (CO) were used. In this study, co-amplification of HV1 and cytb was carried out in order to check the contamination of animal DNA and to verify the human DNA. The primer sets used in PCR were H15997/L16236 for HV1 and H14724/L15149 for cytb. PCR products for HV1 and cytb were 239 bp and 425 bp, respectively. The appearance of two bands on agarose gel implied the DNA came from human, however the single band of cytb gene represented the non-human animal DNA.

Detection and Identification of $\beta$-lactamase, Enterotoxin and Other Exotoxins Genes of Staphylococcus aureus by PCR

  • Yoon, Y.H.;Kim, K.I.
    • Asian-Australasian Journal of Animal Sciences
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    • v.16 no.3
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    • pp.425-429
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    • 2003
  • Staphylococcus aureus is a major pathogen for cattle, causing various forms of subclinical and clinical mastitis and could be a causative agent of food poisoning, it produces various superantigenic exotoxins which have a great public health significance. A total of 72 S. aureus clinical isolates from dairy farms located in Kyunggi Province Korea were examined for the species identification by biochemical method, and for the detection of $\beta$-lactamase, enterotoxin and other exotoxins genes by PCR. The results of species identification by biochemical method agreed with those of PCR done with species specific primer STA-AU. $\beta$-lactamase is an enzyme closely associated with the resistance to antibiotic penicillin, which is an important means of treatment of mastitis, all the isolates were positive for the presence of genes encoding $\beta$-lactamase, which were reproduced in penicillin susceptibility disc assay. Six types of toxin genes, Staphylococcal enterotoxin (SE)A, SEB, SEC, SEE, toxic shock syndrome toxin (TSST-1) and exfoliative toxin A (ET A) were detected in 72 isolates by PCR associated genotypic method in this study, none of the isolates carried the genes for enterotoxin D (SED) and exfoliative toxin B (ETB). The occurrence rate of exotoxin genes rated as 12.5%, and the precision of the PCR identification results has been confirmed using the reference strains.

Identification Based on Computational Analysis of rpoB Sequence of Bacillus anthracis and Closely Related Species (Bacillus anthracis와 그 유연종의 rpoB 유전자 컴퓨터 분석을 통한 동정)

  • Kim, Kyu-Kwang;Kim, Han-Bok
    • Korean Journal of Microbiology
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    • v.44 no.4
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    • pp.333-338
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    • 2008
  • Computational analysis of partial rpoB gene sequence (777 bp) was done in this study to identify B. anthracis and its closely related species B. cereus and B. thuringiensis. Sequence data including 17 B. anthracis strains, 9 B. cereus strains, and 7 B. thuringiensis strains were obtained by searching databases. Those sequences were aligned and used for other computational analysis. B. anthracis strains were identificated by in silico restriction enzyme digestion. B. cereus and B. thuringiensis were not segregated by this method. Those sequencing and BLAST search were required to distinguish the two. In actual identification tests, B. anthracis strains could be identified by PCR-RFLP, and B. cereus and B. thuringiensis strains were distinguished by BLAST search with reliable e-value. In this study fast and accurate method for identifying three Bacillus species, and flow chart of identification were developed.

Development of a Species Identification Method for the Egg and Fry of the Three Korean Bitterling Fishes (Pisces: Acheilognathinae) using RFLP (Restriction Fragment Length Polymorphism) Markers (제한절편 길이 다형성(RFLP) 분자마커를 이용한 납자루아과 담수어류 3종의 난과 치어 종 동정 기법 개발)

  • Choi, Hee-kyu;Lee, Hyuk Je
    • Korean Journal of Environmental Biology
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    • v.36 no.3
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    • pp.352-358
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    • 2018
  • This study aimed to develop a species identification method for the egg and fry of the three Korean bitterling fishes (Pisces: Acheilognathinae), including Acheilognathus signifer, Acheilognathus yamatsutae and Rhodeus uyekii based on the PCR-based Restriction Fragment Length Polymorphism (RFLP) markers. We conducted a field survey on the Deokchicheon River from the North Han River basin, where the three Acheilognathinae species co-occur, and also analyzed the existing sequence dataset available from the GenBank. We found coexistence of the three species at the study site. The egg and fry were obtained from the host mussels (Unio douglasiae sinuolatus) by hand from May to June 2015 and in May 2017. To develop PCR-based RFLP markers for species identification of the three Acheilognathinae fish species, restriction enzymes pinpointing species-specific single nucleotide variation (SNV) sites in mitochondrial DNA COI (cytochrome oxidase I) and cyt b (cytochrome b) genes were determined. Genomic DNA was extracted from the egg and fry and RFLP experiments were carried out using restriction enzymes Apal I, Stu I and EcoR V for A. signifer, A. yamatsutae and R. uyekii, respectively. Consequently, unambiguous discrimination of the three species was possible, as could be seen in DNA band patterns from gel electrophoresis. Our developed PCR-based RFLP markers will be useful for the determination of the three species for the young and would assist in studying the spawning patterns and reproductive ecology of Acheilognathinae fishes. Furthermore, we believe the obtained information will be of importance for future maintenance, management and conservation of these natural and endangered species.

Rapid and exact molecular identification of the PSP (paralytic shellfish poisoning) producing dinoflagellate genus Alexandrium

  • Kim, Choong-jae;Kim, Sook-Yang;Kim, Kui-Young;Kang, Young-Sil;Kim, Hak-Gyoon;Kim, Chang-Hoon
    • Proceedings of the Korean Aquaculture Society Conference
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    • 2003.10a
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    • pp.132-133
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    • 2003
  • The marine dinoflagellate genus Alexandrium comprise PSP producing A. acatenella, A. angustitabuzatum, A. catenella, A. fundyense, A. minutum, A. ostenfezdii, A. tamiyavanichii and A. tamarense. In monitoring toxic Alexandrium, rapid and exact species identification is one of the significant prerequisite work, however we have suffered confusion of species definition in Alexandrium. To surmount this problem, we chose DNA probing, which has long been used as an alternative for conventional identification methods, primarily relying on morphological approaches using microscope in microbial field. Oligonucleotide DNA probes targeting rRNA or rDNA have been commonly used in diverse studies to detect and enumerate cells concerned as a culture-indetendent powerful tool. Despite of the massive literature on the HAB species containing Alexandrium, application of DNA probing for species identification and detection has been limited to a few documents. DNA probes of toxic A. tamarense, A. catenella and A. tamiyavanichii, and non-toxic A. affine, A. fraterculus, A. insuetum and A. pseudogonyaulax were designed from LSU rDNA D1-D2, and applied to whole cell-FISH. Each DNA probes reacted only the targeted Alexandrium cells with very high species-specificity within Alexandrium. The probes could detect each targeted cells obtained from the natural sea water samples without cross-reactivity. Labeling intensity varied in the growth stage, this showed that the contents of probe-targeted cellular rRNA decreased with reduced growth rate. Double probe TAMID2S1 achieved approximately two times higher fluorescent intensity than that with single probe TAMID2. This double probe did not cross-react with any kinds of microorganisms in the natural sea waters. Therefore we can say that in whole-cell FISH procedure this double DNA probe successfully labeled targeted A. tamiyavanichii without cross-reaction with congeners and diverse natural bio-communities.

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Fiber Identification via the TISS and DELTA Systems (TISS system 및 DELTA system에 의한 섬유식별)

  • 전수경
    • Journal of the Korea Furniture Society
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    • v.10 no.1
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    • pp.1-12
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    • 1999
  • Of the vast number of plant taxa in the world, the wood is one of the most useful resources. It is important to identify the fibers of wood and pulp for the plant taxonomy and for the uses, but we do not have enough information on them, on them, especially for the computerizd data. The fiber identification is one of the difficult tasks. In addition to the plant taxonomy and the fiber-using industries, such identification is also important in many other fields, including education. document examiners, etc. For these purpose, the fibers should be exactly distinguished. The TISS system I have programed to identify various woods would also be useful in the identification of fibers by the genus and species in the features of unknown samples and in searching the features of a species based on its scientific name. Such searching programs are being developed in many other countries with a view to searching for the species name by using the features of the cells of the woody materials. With the survey of all the available literature, the features of the fibers of 124 species both of softwood and hardwood were examined under the electron and optical microscopies. Each species were coded and carded by the feature, and the databases were built. The microscopic were inputted into a personal computer program called and by a slide film scanner. The new computer program called TISS 2 was developed using C computer language. Korean language fonts were added to the TISS 2. The TISS 2 can be in adding and searching a image of fiber features both of a known fiber and an unknown fiber. The databases were corded for the DELTA system with was developed by Dallwitz and Paine in Australia, 1986.

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Molecular Sexing and Species Identification of the Processed Meat and Sausages of Horse, Cattle and Pig

  • Kim, Yoo-Kyung;Kang, Yong-Jun;Kang, Geun-Ho;Seong, Pil-Nam;Kim, Jin-Hyoung;Park, Beom-Young;Cho, Sang-Rae;Jeong, Dong Kee;Oh, Hong-Shik;Cho, In-Cheol;Han, Sang-Hyun
    • Journal of Embryo Transfer
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    • v.31 no.1
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    • pp.61-64
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    • 2016
  • We developed a polymerase chain reaction (PCR)-based molecular method for sexing and identification using sexual dimorphism between the Zinc Finger-X and -Y (ZFX-ZFY) gene and polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) for mitochondrial DNA (mtDNA) cytochrome B (CYTB) gene in meat pieces and commercial sausages from animals of different origins. Sexual dimorphism based on the presence or absence of SINE-like sequence between ZFX and ZFY genes showed distinguishable band patterns between male and female DNA samples and were easily detected by PCR analyses. Male DNA had two PCR products appearing as distinct two bands (ZFX and ZFY), and female DNA had a single band (ZFX). Molecular identification was carried out using PCR-RFLP of CYTB gene, and showed clear species classification results. The results yielded identical information on the sexes and the species of the meat samples collected from providers without any records. The analyses for DNA isolated from commercial sausage showed that pig was the major source but several sausages originated from chicken and Atlantic cod. Applying this PCR-based molecular method was useful and yielded clear sex information and identified the species of various tissue samples originating from livestock.

An External and Micromorphological Identification for Pharbitidis Semen and its Congeneric Species (외부 및 미세형태 비교를 통한 견우자(牽牛子) 기원종 및 동속이종(同屬異種) 감별)

  • Song, Jun-Ho;Yang, Sungyu;Choi, Goya;Moon, Byeong Cheol
    • The Korea Journal of Herbology
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    • v.33 no.4
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    • pp.43-51
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    • 2018
  • Objectives : Pharbitidis Semen, the seeds of Ipomoea nil (L.) Roth or I. purpurea (L.) Roth, is well-known traditional herbal medicine in Korea. But it is often marketed as a different seed or mixtures of its closely related species. Thus, the present study aims to provide external and micromorphological characters and identification key by using stereoscope (ST) and scanning electron microscope (SEM) for discriminating authentic of Pharbitidis Semen. Methods : A discrimination on external morphological characteristics of sepals, fruits, seeds, and hilum, testa cell micromorphology in the original plants and its congeneric species was carried out using digital calipers, ST, and SEM. Results : Number of valves (degree of apex of each valve), number of seeds per locule, hairy in capsules and size, luster, density of hairy, hilum shape in seeds and shape of cell, anticlinal, periclinal wall in testa may have high discriminative value. The seeds of Ipomoea nil as an original plant of Pharbitidis Semen were distinguished from other species by the relative larger in size, ovoid-trigonous in shape, mostly flabellate or triangular to trapezoid in outline (c.s.), dull, and puberulent in surface and thicken anticlinal wall. Conclusions : On the basis of the results, an identification key of Pharbitidis Semen and closely related species is provided. Our observations suggest that the combination of morphological characters and other studied results could be helpful in the successfully identified authentic herbal medicines. Moreover, micromorphological characters using SEM could be useful for discriminating authentic medicines.

A study on the fine structure of marine diatoms in Korean coastal waters: Genus Thalassiosira 5

  • Park, Joon-Sang;Lee, Jin-Hwan
    • ALGAE
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    • v.25 no.3
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    • pp.121-131
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    • 2010
  • Thalassiosira species were collected from October 2007 to January 2009 in an attempt to better understand species diversity of the genus Thalassiosira in Korean coastal waters. A total of 5 Thalassiosira species (T. concaviuscula, T. oceanica, T. partheneia, T. simonsenii and T. nanolineata) were identified here. Most species in this study were of small size, and 5 species were recorded for the first time in Korean coastal waters. Using a scanning electron microscope (SEM), we described distinctive characteristics of fine structure that proved to be important diagnostic characteristics for the identification of each species. The most important diagnostic characteristics for Thalassiosira species identification were the marginal strutted processes, the position of labiate processes, and the areolation. The differential characteristics of the species studied were: T. concaviuscula has a double layered external tubes on the marginal strutted processes; T. oceanica shows marginal ridges that are interlinked between the marginal strutted processes; the valve face of T. partheneia is fairly convex and its labiate process is positioned midway between two strutted processes; T. simonsenii is characterized by two labiate processes and somewhat coarse areolae; and, T. nanolineata has several central strutted processes and linear areolation.