• Title/Summary/Keyword: Species-specific PCR primer

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Spread of Cyst Nematodes in Highland Chinese Cabbage Field in Gangwon-do (강원도 고랭지배추 재배지에서 씨스트선충의 분포 확산)

  • Kwon, Soon-Bae;Park, Dong-Kwon;Won, Heon-Seop;Moon, Youn-Gi;Lee, Jae-Hong;Kim, Yong-Bog;Choi, Byoung-Gon;Seo, Hyun-Taek;Ko, Hyoung-Rai;Lee, Jae-Kook;Lee, Dong Woon
    • Korean journal of applied entomology
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    • v.57 no.4
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    • pp.339-345
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    • 2018
  • The sugar beet cyst nematode (SBCN), Heterodera schachtii first detected in Taebaek, Gangwon-do in 2011, is one of the major plant parasitic nematodes that cause economic damage to the Chinese cabbage in highland regions. In addition, the distribution of clover cyst nematode (CCN), H. trifolii was confirmed in the highland Chinese cabbage cultivated regions in 2017. In order to investigate the spread of cyst nematodes, this study has been conducted since 2013 in the highland Chinese cabbage cultivation area. In addition, in 2017, the Real-Time PCR technique with the species-specific primer was used to investigate those two cyst nematodes and the soybean cyst nematode (SCN), H. glycines which is known for its distribution in Korea, focusing on the main production regions of highland Chinese cabbage cultivation. The number of infected fields in the Chinese cabbage plantation in highland increased every year to confirm distribution in Taebaek, Samcheok, Jeongseon and Gangneung in 2017, and the cumulative number of infection fields reached 245 by 2017. Of the 41 possible cyst nematode samples for PCR analysis, 61% were CCN, only 9.8% of the SBCN and 29.3% of the SCN were identified. Therefore, some of the previously known SBCN or CCN discoveries are likely to have been infected with SCN. It is believed that the CCN needs to be controlled in the future as CCN have been found to be dominant species in the highland Chinese cabbage plantation regions.

Characterization of Three Korean Isolates of Malva Vein Clearing Virus from Curled Mallow (Malva verticillata) (아욱에서 분리한 Malva Vein Clearing Virus 분리주의 특성)

  • Kwak, Hae-Ryun;Kim, Ji-Gwang;Kim, Jeong-Eun;Choi, Hyeon-Yong;Choi, Hong-Soo;Kim, Mikyeong
    • Research in Plant Disease
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    • v.26 no.4
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    • pp.283-288
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    • 2020
  • In September 2017, vein clearing and yellowing symptoms resembling those caused by viruses were observed on leaves of Malva verticillata in Chungnam, Korea. Nucleic acids were extracted from leaves of five symptomatic plants and tested by reverse transcription polymerase chain reaction using four virus specific primer pairs including malva vein clearing virus (MVCV). Amplicons of the expected size (600 bp) were obtained from total RNA of all samples using the MVCV-specific primers. To confirm the presence of MVCV in symptomatic plants, the DNA fragments from three samples were purified, and directly sequenced. BLAST analysis revealed that it shared the highest nucleotide identity (99%) with a MVCV isolate from tomato (Mexico). The virus isolates obtained from the third re-inoculated Chenopodium was designated as Cm1-5. Tissue from Cm1, Cm3, and Cm5 isolates was mechanically sap inoculated into 23 indicator plants. Cm3 isolate induced chlorotic local and mosaic symptoms in Althaea rosea. Phylogenetic analysis based on coat protein gene of 19 MVCV isolates from 6 different countries and plant species, did not correlated with either the geographical origin of the isolates, or pathogenicity. To our knowledge, this study first reports the natural occurrence of MVCV on M. verticillata in Korea and characterization of three Korean isolates of MVCV.

DETECTION OF METHICILLIN OR VANCOMYCIN-RESISTANT STAPHYLOCOCCUS AUREUS FROM DENTAL HOSPITAL (치과병원 진료실 내에서 메티실린 또는 반코마이신 저항성 Staphylococcus aureus의 검출)

  • Min, Jung-Hee;Park, Soon-Nang;Hwang, Ho-Keel;Min, Jung-Beum;Kim, Hwa-Sook;Kook, Joong-ki
    • Restorative Dentistry and Endodontics
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    • v.32 no.2
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    • pp.102-110
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    • 2007
  • The purpose of this study was to obtain the basic information for the improvement of dental environment by investigating the presence of methicillin- or vancomycin-resistant Staphylococcus aureus (MRSA or VRSA) isolated from dental health care workers (DHCWs) and environment of the Chosun University Dental Hospital (CUDH) and a private dental clinic (control group). Staphylococcus aureus (S. aureus) was isolated from anterior nares of 42 DHCWS and 38 sites, unit chairss, x-ray devices, computers, etc., at 10 departments of the CUDH and 20 DHCWs and 11 sites at the private dental clinic. S. aureus was isolated on mannitol salt agar plate and confirmed by PCR with S. aureus species-specific primer. Antimicrobial susceptibility test of clinical isolates of S. aureus against several antibiotics including methicillin (oxacillin) was performed by investigating minimum inhibitory concentration (MIC) using broth microdilution assay. In addition, PCR was performed to detect the methicillin- or vancomycin-resistant gene. The data showed that one strain of S. aureus was isolated from DHCWs of the CUDH and three strains of S. aureus was isolated from 3 samples of the private dental clinic, respectively. All of the isolates from the CUDH and the private dental clinic had resistance to penicillin G, amoxicillin and vancomycin and susceptibility to oxacillin and ciprofloxacin. The S. aureus strains were already obtained the resistance to penicillin G and amoxicillin. These results suggest that two dental clinics were under relatively safe environment.

Screening and Characterization of LTR Retrotransposons in the genomic DNA of Pleurotus eryngii (큰느타리버섯 유전체내 LTR Retrotransposon 유전자 탐색 및 특성연구)

  • Kim, Sinil;Le, Quy Vang;Kim, Sun-Mi;Ro, Hyeon-Su
    • The Korean Journal of Mycology
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    • v.42 no.1
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    • pp.50-56
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    • 2014
  • Transposable elements (TEs) are mobile DNA elements that often cause mutations in genes and alterations in the chromosome structure. In order to identify and characterize transposable elements (TEs) in Pleurotus eryngii, a TE-enriched library was constructed using two sets of TE-specific degenerated primers, which target conserved sequences of RT and RVE domains in fungal LTR retrotransposons. A total of 256 clones were randomly chosen from the library and their insert sequences were determined. Comparative investigation of the insert sequences with those in repeat element database, Repbase, revealed that 71 of them were found to be TE-related fragments with significant similarity to LTR retrotransposons from other species. Among the TE sequences, the 70 TEs were Gypsy-type LTR retrotransposons, including 20 of MarY1 from Tricholoma matsutake, 26 of Gypsy-8_SLL from Serpula lacrymans, and 16 of RMER17D_MM from mouse, whereas a single sequence, Copia-48-PTR, was found as only Copia-type LTR retrotransposon. Southern blot analysis of the HindIII-digested P. eryngii genomic DNA showed that the retrotransposon sequences similar to MarY1 and Gypsy-8_SLL were contained as high as 14 and 18 copies per genome, respectively, whereas other retrotransposons were remained low. Moreover, both of the two Gypsy retrotransposons were expressed in full length mRNA as shown by Northern blot analysis, suggesting that they were functionally active retrotransposons.

Expression of Cytochrome P450 Aromatase Genes during Sex Differentiation in Korean Rockfish, Sebastes schlegeli (조피볼락, Sebastes schlegeli의 성분화 기간 중 Cytochrome P450 Aromatase 유전자의 발현)

  • Lee, Chan-Hee;Kwon, Joon-Yeong
    • Development and Reproduction
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    • v.11 no.3
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    • pp.195-203
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    • 2007
  • Sex determination and sex differentiation are influenced by genotype in many gonochoristic fish. Cytochrome P450 aromatase (CYP19) is the terminal enzyme in steridogenic pathway that converts androgens into estrogens. In this study, partial fragments of aromatase genes (ovarian aromatase, P450aromA and brain aromatase, P450aromB) were cloned and sequenced in Korean rockfish (Sebastes schlegeli), and gene specific primers were designed based on their sequences. Using these primers, aromatase gene expression during sex differentiation was investigated by RT-PCR. Expression of these aromatase genes were detected both in the head and body parts at 35 dab (days after birth). The number of fish that expressed the aromatase genes decreased at 52 dab, implying down-regulation of these genes. However, these genes were expressed at 59 dab in almost all fish studied here. The expression patterns of both genes are similar throughout the investigated period except for 45 dab where the expression of P450aromB was detected in more fish than that of P450aromA both in the head and body parts. Timing of sex differentiation in this species has been shown to be at around $50{\sim}65$ dab by histological analysis. However, the results from this study suggest that sex differentiation of rockfish may take place $1{\sim}2$ weeks earlier than the period proposed previously. The results also suggest that the mechanism of sex differentiation in viviparous fish may be similar to that in oviparous fish in terms of the importance of aromatase action during the critical period.

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Identification of a New Potyvirus, Keunjorong mosaic virus in Cynanchum wilfordii and C. auriculatum (큰조롱과 넓은잎 큰조롱에서 신종 포티바이러스(큰조롱모자이크바이러스)의 동정)

  • Lee, Joo-Hee;Park, Seok-Jin;Nam, Moon;Kim, Min-Ja;Lee, Jae-Bong;Sohn, Hyoung-Rac;Choi, Hong-Soo;Kim, Jeong-Soo;Lee, Jun-Seong;Moon, Jae-Sun;Lee, Su-Heon
    • Research in Plant Disease
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    • v.16 no.3
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    • pp.238-246
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    • 2010
  • In 2006 fall, a preliminary survey of viruses in two important medicinal plants, Cynanchum wilfordii and C. auriculatum, was conducted on the experimental fields at the Agricultural Research and Extension Services of Chungbuk province in Korea. On each experimental fields, percentage of virus infection was ranged from 20 to 80%, and especially an average of disease incidence propagated by roots was twice higher than that by seeds. The various symptoms were observed in Cynanchum spp. plants, such as mosaic, mottle, necrosis, yellowing, chlorotic spot and malformation etc. In electron microscopic examination of crude sap extracts, filamentous rod particles with 390-730 nm were observed in most samples. The virus particles were purified from the leaves of C. wilfordii with typical mosaic symptom, and the viral RNA was extracted from this sample containing 430-845 nm long filamentous rod. To identify the viruses, reverse transcription followed by PCR with random primers was carried out. The putative sequences of P3 and coat protein of potyvirus were obtained. From a BLAST of the two sequences, they showed 26-38% and 62-72% identities to potyviruses, respectively. In SDS-PAGE analysis, the subunit of coat protein was approximately 30.3 kDa, close to the coat protein of potyvirus. In bioassay with 21 species in 7 families, Chenopodium quinoa showed local lesion on inoculated leave and chlorotic spot on upper leave, but the others were not infected. RT-PCR detection using specific primer of C. wilfordii and C. auriculatum samples, all of 24 samples with virus symptom was positive, and five out of seven samples without virus symptom were also positive. On the basis of these data, the virus could be considered as a new member of potyvirus. We suggested that the name of the virus was Keunjorong mosaic virus (KjMV) after the common Korean name of C. wilfordii.

Microbiological Quality of Raw and Cooked Foods in Middle and High School Food Service Establishments (서울시 일부 중.고등학교의 급식용 식재료 및 조리식품의 미생물학적 품질)

  • Kim, Myung-Hee;Shin, Weon-Sun
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.37 no.10
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    • pp.1343-1356
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    • 2008
  • The evaluation of microbiological quality for school food samples collected from 19 selected middle and high schools located in Seoul was undertaken. Eighty-nine food samples consisting of 38 non-pretreated vegetables, 13 pre-washed and cut vegetables, 9 meats and poultry, 3 fish and shellfish, 7 dried fish, and shellfish and 20 processed foods were collected. Aerobic plate count, total coliforms, and Escherichia coli (E. coli ) were detected using $Petrifilm^{TM}$, and the food-borne pathogens were screened by multiplex PCR with species-specific primer sets. Sequentially, the quantitative and confirmative test of the food-borne pathogens were carried out with the selective media and biochemical kits. The contamination of coliform counts was observed on the pre-washed vegetables ($3.4{\sim}4.3\;log\;CFU/g$) and meats ($2.2{\sim}4.3\;log\;CFU/g$). Also, the cooked foods were heavily contaminated with coliform, ranging from 1.0 to $5.5\;log\;CFU/g$. E. coli counts were found in 16 raw and cooked food samples, exceeding the microbiological standards for the guideline of safety management for school foods. Through PCR detection, B acillus cereus was detected in 32 raw and cooked foods, and quantitatively found in pre-washed carrot, radish, and pan-broiled dried shrimp and filefish ranging from $2.3{\sim}3.6\;log\;CFU/g$, respectively. E. coli O157:H7 was detected on frozen pork sample and was confirmed with API kit. Campylobacter jejuni was found in 3 ready-to-eat type vegetables. Vibrio parahaemolyticus were found in 4 pre-washed vegetables and 2 cooked foods, indicating unsatisfactory quality based upon the microbiological standards of ready-to-eat vegetables and cooked foods by Korea Food and Drug Administration. Salmonella spp. was detected in frozen chicken sample and confirmed by API kit and latex antisera agglutination.