• Title/Summary/Keyword: 바이러스 유전자 검정

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Cloning of Coat Protein Gene from Korean Isolate Potato Leafroll Virus (PLRV) and Introduction into Potato (Solanum tuberosum) (한국 분리주 감자 잎말림 바이러스 (PLRV) 외피 단백질 유전자의 클로닝 및 감자 내 도입)

  • Seo Hyo-Won;Yi Jung-Yoon;Park Young-Eun;Cho Ji-Hong;Hahm Young-Il;Cho Hyun-Mook
    • Journal of Plant Biotechnology
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    • v.32 no.4
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    • pp.243-250
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    • 2005
  • The coat protein gene (AF296280) of the Korean isolate Potato leafroll virus (PLRV) was cloned and the open reading frame (627 bp) was transformed into potato (Solanum tuberosum cv. Superior). Out of seventeen individual transgenic lines, five lines were identified to confer resistance to PLRV through the five generation's selection program in the greenhouse as well as isolated trial field. Successful introduction and genetic stability of coat protein gene in the genome of potato were confirmed by polymerase chain reaction (PCR), Southern blot hybridization and northern blot hybridization. Some of the transgenic lines were highly resistant to PLRV but did not show any resistance to less homologous Potato virus Y (PVY). Our results suggest that the resistance to PLRV is due to homology dependent gene silencing by sense strand coat protein gene. In addition, the results of field test through five generations showed that there were no significant differences comparing to nontransgenic potatoes in the morphological aspect of shoot as well as tuber, Ho remarkable differences were also observed in the major agronomic characters and yields except for the resistance to PLRV.

Analysis and Detection of Coast Protein Gene of Barley Yellow Mosaic Virus and Barley Mield Mosaic Virus by RT-PCR (RP-PCR을 이용한 보리누른모자이크바이러스 (BaYMV)와 보리마일드모자이크바이러스(BaMMV)의 외피단백질 유전자 검정 및 해석)

  • 이귀재
    • Korean Journal Plant Pathology
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    • v.14 no.4
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    • pp.314-318
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    • 1998
  • Using the reverse transcription polymerase chain reaction (RT-PCR), a rapid and sensitive assay method for the detection and identification of barley yellow mosaic virus (BaYMV) and barley mild mosaic virus (BaMMV) was adapted. Two units of primers from each virus were selected and used for the determination of two different viruses. PCR fragments of BaYMV (ca. 0.9kb) and BaMMV (ca. 0.8kb) were obtained from the designed method for the assay of BaYMV and BaMMV coat protein. PT-PCR fragments were cloned using vector pT7 Blue and the sequences of the selected clones were analyzed. coat protein of BaYMV and that of BaMMV consisted of 297 amino acids (891 nucleotides) and 251 amino acids (753 nucleotides), respectively. The snalysis of coat protein genes from these two viruses showed that 45.6% of nucleotides sequence ad 34.9% of amino acid in BaYMV were homologous to those in BaMMV.

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Screening of Tomato Spotted Wilt Virus Resistance in Tomato Accessions (토마토반점위조바이러스(TSWV) 저항성 토마토 유전자원 탐색)

  • Han, Jung-Heon;Choi, Hak-Soon;Lee, Jun-Dae;Kim, Jae-Deok;Lee, Won-Phil;Choi, Hong-Soo;Kim, Jung-Soo;Yoon, Jae-Bok
    • Horticultural Science & Technology
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    • v.30 no.2
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    • pp.171-177
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    • 2012
  • A total of 94 tomato accessions were evaluated for the resistance to $Tomato$ $spotted$ $wilt$ $virus$ (TSWV) using a Sw5-2 SCAR marker and bioassay. PCR products of the marker were approximately 574 bp, 500 bp, and 462 bp, among which the longest was linked to TSWV resistance allele of Sw5-b. This allele was only found in three accessions (09-438, 10-318, and 10-321) in which some individuals showed apparent recovery or stem necrosis symptom to a tomato isolate of TSWV-pb1. Thirty-five individuals (one per each accession) which were non-infected by ELISA were selected for further observation. Among these, 26 individuals that did not show any symptom at 5 months after inoculation were confirmed for viral infection by RT-PCR. TSWV-specific PCR amplicon was weakly detected in all 26 individuals including 'Eureta', a commercial F1 possessing the resistance allele of Sw5-b. The resistant genes in the selected individuals may play an important role for reducing the viral concentration in tissues of inoculated tomato plants and seems to be quantitatively controlled by several factors including Sw5-b gene.

Resistance Evaluation of Commmercial Tomato Cultivars against Tomato yellow leaf curl virus (토마토품종의 토마토황화잎말림바이러스병에 대한 저항성 평가)

  • Ko, Sug-Ju;Kim, Hyo-Jeong;Lee, Jin-Hee;Ma, Kyung-Cheol;Choi, Duck-Soo;Park, Young-Hoon;Choi, Seung-Kook;Kim, Mi Kyeong;Choi, Hong-Soo
    • Research in Plant Disease
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    • v.22 no.4
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    • pp.297-302
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    • 2016
  • Tomato yellow leaf curl virus (TYLCV) is a viral disease causing severe economic losses on tomato. Practical prevention of the TYLCV disease is to control tabacco whitefly (Bemisia tabaci) or to cultivate TYLCV-resistant tomato cultivars, because no agrochemical products are available to control TYLCV. In this study, TYLCV resistance of the commercial tomato cultivars were evaluated using the DNA markers tightly linked to TYLCV resistance genes Ty-1 and Ty-3 and infection with the TYLCV clones mediated by Agrobacterium. In marker genotyping, resistance alleles were detected from 4 oval type tomato cultivars (Titichal, TY tinny, TY saengsaeng II, TY sense Q). Four cheery type cultiavrs (TY endorphin, TY smartsama, Tiara TY, Olleh TY) and 6 round type cultivars (TY kingdom, TY ace, TY homerun, TY altorang, Dotaerang TY winner, Styx TY). The seedling bioassay indicated that tomato cultivars of the oval type and cherry type showed consistancy in marker genotype and phenotype while slight disease symptom was observed from some round type cultivras (TY ace, TY homerun, Styx TY) with resistance marker genotype. For fruit yields, TY tinny was greater than its control cultivar Titichal in oval types, TY smartsama was greater than its control Smile in cherry type, and TY ace and TY kingdom were greater than their control Dabok. These cutliavrs can be a good choice for high-yielding TYLCV-resistant tomato cultivars.

Detection of Cucumber green mottle mosaic virus in Bottle Gourd Seeds by RT-PCR (RT-PCR에 의한 박 종자의 오이녹반모자이크바이러스 검정)

  • Lee, Sook-Kyung;Song, Wan-Yeob;Kim, Hyung-Moo
    • Research in Plant Disease
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    • v.10 no.1
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    • pp.53-57
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    • 2004
  • Cucumber green mottle mosaic virus (CGMMV) was a major pathogen of watermelon and had affected seriously to watermelon production in Korea. Rapid and sensitive detection method of CGMMV associated with bottle gourd (Lagenafia siceraria) seeds was developed by using RT-PCR in this study. A pair of primeri Wmfl and Wmrl, specific for CGMMV was designed from coat protein gene sequences of CGMMV-W and used for amplifying 420 bp product in RT-PCR. To simplify the virus extraction procedure and reduce an inhibitor from the extract for the RT-PCR, some methods using ethanol precipitation, double filtration, polyethylene glycol precipitation and phenol/chloroform/isoamyl alcohol extraction procedure were compared and the phenol/chloroform/isoamyl alcohol extraction procedure was selected by its enhanced sensitivity. This detection method using the selected extraction step and the primers for RT-PCR could reliably detect an infected level of one CGMMV-infested seed in 1,000 seeds. This rapid and sensitive RT-PCR assay provides auseful tool for the specific detection of CGMMV in bottle gourd seed samples containing high levels of back-ground inhibitors.

Incidence and Occurrence Pattern of Viruses on Peppers Growing in Fields in Korea (국내 노지재배 고추의 바이러스 발생률 및 발병 현황)

  • Kwon, Sun-Jung;Cho, In-Sook;Yoon, Ju-Yeon;Chung, Bong-Nam
    • Research in Plant Disease
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    • v.24 no.1
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    • pp.66-74
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    • 2018
  • Field surveys to investigate the incidence and occurrence pattern of viruses in red pepper were conducted during 2015-2016 in Korea. A total of 424 samples in 2015 and 368 samples in 2016 were collected based on selection of plants showing symptoms from farmer's field from every June to September. Reverse transcription-Polymerase chain reaction was used to test all samples for the presence of one or more of following viruses: Cucumber mosaic virus (CMV), Broad bean wilt virus 2 (BBWV2), Tomato spotted wilt virus (TSWV), Beet western yellows virus (BWYV), Pepper mottle virus (PepMoV), Potato virus Y (PVY) and Pepper mild mottle virus (PMMoV). The average disease incidence was 91.7% in 2015 and 98% in 2016 and the all seven viruses were found although there were different kinds of regions. The percent virus incidence in collected samples during 2015 was as follows: CMV, 73.8%; BBWV2, 68.3%; BWYV, 46.9%; PMMoV, 14.6%; TSWV, 12.7%; PepMoV, 6.6% and PVY, 3.3%. For 2016, incidence was as follows: CMV, 73.3%; BBWV2, 71.4%; BWYV, 34.7%; TSWV, 27.9%, PMMoV, 19.2%; PepMoV, 13.5% and PVY, 3.5%. Mixed infections were prevalent over single infections and infection rate was 83% and 86.7% in 2015 and 2016, respectively.

An Effective Detection of Potato Virus Y Using RT-PCR Technique (RT-PCR 기법을 이용한 효과적인 감자바이러스 Y의 검정)

  • Joung, Young-Hee;Jeon, Jae-Heung;Choi, Kyung-Hwa;Kim, Hyun-Soon;Yi, Yong-Sub;Joung, Hyouk
    • Korean Journal Plant Pathology
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    • v.13 no.4
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    • pp.219-224
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    • 1997
  • A PT-PCR (reverse transcription-polymerase chain reaction) diagnostic method for potato virus Y (PVY) was developed using primer pair derived from conserved region of coat protein genes of several PVY strains, A 764 bp PCR product was detected from several lines of potato cv. Atlantic. We could prove that the 764 bp DNA fragment was indeed the PVY gene by sequencing analysis. PVY detection method using RT-PCR technique was about tuber tissue.

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Incidence of Virus Diseases in Major Cultivated Areas of Watermelon and Melon in Chungbuk Province (충북지역 주산지 수박, 멜론에서의 바이러스 발생현황)

  • Jong-Woo Han;Young-Uk Park;Cheol-Ku Youn;Seok-Ho Lee;Taek-Goo Jeong;Hong-Soo Choi;Mi-Kyeong Kim
    • Research in Plant Disease
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    • v.29 no.1
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    • pp.88-93
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    • 2023
  • To investigate the incidence status of viruses in major cultivated areas of watermelon and melon in Chungbuk Province, samples were collected from 2020 to 2021 in vinyl greenhouse of Jincheon and Eumseong and examined for virus infection using reverse transcription polymerase chain reaction. Of the six viruses on watermelon that was analyzed in this study, watermelons were infected with cucumber mosaic virus (CMV), watermelon mosaic virus (WMV), cucumber green mottle mosaic virus (CGMMV), and cucurbit aphid-borne yellows virus (CABYV). The incidence rate of CMV was 20.9-35.0%, WMV 0.4-15.8%, CGMMV 1.6-38.5%, and CABYV was 3.5-3.7% from 2020 to 2021. But strangely, there were no incidence of zucchini yellow mosaic virus and cucurbit chlorotic yellows virus (CCYV) during investigation. From this result, we knew the major virus was CGMMV on watermelon in Chungbuk Province. Molecular diagnosis assays of the two melon viruses, showed that melons were infected with CABYV and CCYV from 2020 to 2021. The incidence rate of CABYV was 53.9-92.2% and CCYV was 2.7-20.8%. The incidence of CABYV was high in melon cultivation of Jincheon and Eumseong, Chungbuk. Afterwards, it is necessary to establish a control management strategy for reduce the incidence of CABYV. Furthermore, we must pay attention that of CCYV even if the incidence was low.

Survey of the Routes and Incidence of Viral Infection of Tomato and Paprika Growing in Greenhouses in Cherwon Province, Korea During 2015-2017 (2015-2017 철원군 시설재배 과채류(토마토와 파프리카)의 바이러스병 발생 현황 및 감염경로 조사)

  • Kil, HyungBae;Kang, Minji;Choi, Won-Seok;Kim, Joong-Il;Phan, Mi Sa Vo;Im, JiHui;Kim, MeeKyoung;Park, Mi-Ri
    • Research in Plant Disease
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    • v.24 no.2
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    • pp.145-152
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    • 2018
  • During 2015-2017, we surveyed the incidence of viral infections of tomato and paprika growing in greenhouses in Cherwon province, Korea. In 2015 and 2016, we collected leaves and fruits from tomato and paprika plants growing in greenhouses. We detected viruses in the samples collected using specific primer sets for Broad bean wilt virus 2 (BBWV2), Cucumber mosaic virus (CMV), Pepper mild mottle virus (PMMoV), Pepper mottle mosaic virus (PepMoV), and Tomato spotted wilt virus (TSWV). We detected PMMoV, CMV, and TSWV in the samples, and CMV and TSWV were the most prevalent. For the prevention of future viral diseases, we then surveyed the routes of infection by these viruses in tomato and paprika plants growing in greenhouses in Cherwon province in 2017. Leaf and fruit samples were collected from seedlings and crops two and four months after transplanting into greenhouses. As a result, we found that TSWV was transferred from seedlings to plants, and outbreaks of the virus occurred at the early stage of cultivation. On the other hand, we found that CMV was a virus indigenous to the soil of some towns in Cherwon province, and thus outbreaks of this virus occurred at the middle stage of cultivation.

Characterization of Prunus necrotic ringspot virus Isolate from Peach in Korea (국내 복숭아에서 분리한 Prunus necrotic ringspot virus의 특성)

  • Kim, Hyun-Ran;Lee, Sin-Ho;Shin, Il-Sheob;Kim, Jeong-Hee;Cho, Kang-Hee;Heo, Seong;Kim, Jeong-Soo;Choi, Yong-Mun
    • Research in Plant Disease
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    • v.15 no.3
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    • pp.170-174
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    • 2009
  • In this paper, we report a characterization of Prunus necrotic ringspot virus (PNRSV) isolate. The virus was identified from 'Yumyeong' peach showing mild mosaic on leaves in commercial orchard of 'Umsung', Chungbuk province in Korea. The virus isolate produced ringspot symptom on the inoculated cotyledons and systemic mosaic and malformation on the upper leaves of Cucumis sativus. Systemic mottles were appeared in Chenopodium quinoa. When the buds of the virus infected stem were grafted on the healthy young Prunus persica GF305 seedlings, line pattern with mosaic appeared within 3 months. Isometric virus-like particles were found in parenchyma cells and plasmodesmata of C. sativus leaves inoculated mechanically with the virus. The cDNA fragments of PNRSV coat protein (CP) region, approximately 675bp, were synthesized from genomic RNA extracted from virus-infected leaves by RT-PCR using specific primer pairs. Partial nucleotide sequences of the CP regions were determined and analyzed with the known PNRSV. The CP gene of PNRSVKorea isolates showed 93.9~94.7% similarity to the 4 known PNRSV isolates.