• Title/Summary/Keyword: seed-borne virus

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Immunocapture RT-PCR for Detection of Seed-borne Viruses on Cucurbitaceae Crops (Immunocapture RT-PCR을 이용한 박과작물 종자전염 바이러스의 검출)

  • Lee, Hyok-In;Kim, Jung-Hee;Yea, Mi-Chi
    • Research in Plant Disease
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    • v.16 no.2
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    • pp.121-124
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    • 2010
  • Immunocapture reverse transcription polymerase chain reaction (IC-RT-PCR) was applied to the detection of Cucumber green mottle mosaic virus (CGMMV), Kyuri green mottle mosaic virus (KGMMV), and Zucchini green mottle mosaic virus (ZGMMV) on Cucurbitaceae crops. These seed-borne tobamoviruses were accurately detected from the infected leaves and seeds by IC-RT-PCR. This method was estimated to be about 100 times more sensitive than ELISA, and also it allowed the direct confirmation of ELISA results by using the captured antigens from a completed ELISA microwell. This convenient and reliable method could be used routinely for large-scale field surveys or seed tests of Cucurbitaceae crops.

Development of PCR Diagnosis System for Plant Quarantine Seed-borne Wheat Streak Mosaic Virus (식물검역 종자전염 Wheat Streak Mosaic Virus의 PCR 검사시스템 개발)

  • Lee, Siwon;Kang, Eun-Ha;Chu, Yeon-Mee;Shin, Yong-Gil;Ahn, Tae-Young
    • Korean Journal of Microbiology
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    • v.49 no.2
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    • pp.112-117
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    • 2013
  • Wheat streak mosaic virus (WSMV), a member of the genus Tritimovirus in Potyviridae, severely impacts wheat and corn seed worldwide, but has yet to be detected in Korea, and hence, every effort should be made to prevent its introduction. To prevent WSMV from entering the country, it is necessary to prepare a specific, sensitive, simple, and fast detection method for routine application to plant quarantine procedures. For this reason, a two-step diagnosis system consisting of RT-PCR and nested PCR is being used for WSMV detection. In addition, a novel positive control was developed for use with the system. WSMV has been detected in seed sweet corn from Japan and seed wheat from USA by a two-step diagnosis system, the details of which are described in this study. After sequence analysis, similarities of 80.6 and 100.0% with other isolates were determined by BLAST. They showed the same topology, which was classified as 4 genotypes by various phylogenetic trees, using a poly protein encoding sequence amplification. In this analysis, WSMV-JSweet-corn2868 (JX845574) is classified as clade B, while WSMV-Uwheat1944-1 (KC754959) and WSMV-Uwheat1944-2 (KC754960) belong to clade D.

Characteristics of a Lettuce mosaic virus Isolate Infecting Lettuce in Korea

  • Lim, Seungmo;Zhao, Fumei;Yoo, Ran Hee;Igori, Davaajargal;Lee, Su-Heon;Lim, Hyoun-Sub;Moon, Jae Sun
    • The Plant Pathology Journal
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    • v.30 no.2
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    • pp.183-187
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    • 2014
  • Lettuce mosaic virus (LMV) causes disease of plants in the family Asteraceae, especially lettuce crops. LMV isolates have previously been clustered in three main groups, LMV-Yar, LMV-Greek and LMV-RoW. The first two groups, LMV-Yar and LMV-Greek, have similar characteristics such as no seed-borne transmission and non-resistance-breaking. The latter one, LMV-RoW, comprising a large percentage of the LMV isolates contains two large subgroups, LMV-Common and LMV-Most. To date, however, no Korean LMV isolate has been classified and characterized. In this study, LMV-Muju, the Korean LMV isolate, was isolated from lettuce showing pale green and mottle symptoms, and its complete genome sequence was determined. Classification method of LMV isolates based on nucleotide sequence divergence of the NIb-CP junction showed that LMV-Muju was categorized as LMV-Common. LMV-Muju was more similar to LMV-O (LMV-Common subgroup) than to LMV-E (LMV-RoW group but not LMV-Common subgroup) even in the amino acid domains of HC-Pro associated with pathogenicity, and in the CI and VPg regions related to ability to overcome resistance. Taken together, LMV-Muju belongs to the LMV-Common subgroup, and is expected to be a seed-borne, non-resistance-breaking isolate. According to our analysis, all other LMV isolates not previously assigned to a subgroup were also included in the LMV-RoW group.

One-step Multiplex RT-PCR Method for Simultaneous Detection of Seed Transmissible Bacterium and Virus Occurring on Brassicaceae Crop Seeds (십자화과 작물 종자에서 종자전염 세균 및 바이러스 동시 검출을 위한 One-step Multiplex RT-PCR 방법)

  • Jeong, Kyu-Sik;Soh, Eun-Hee
    • Research in Plant Disease
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    • v.17 no.1
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    • pp.52-58
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    • 2011
  • The aim of this research was to develop specific and sensitive PCR-based procedures for simultaneous detection of economically important plant pathogenic bacteria and seed borne virus in commercial Brassicaceae crop seeds, Xanthomonns campestris pv. campestris (Xcc) and Lettuce Mosaic Virus (LMV). Bacterial and virus diseases of Brassicaceae leaves are responsible for heavy losses. PCR with arbitral primers: selection of specific primers, performance of PCR with specific primers and determination of the threshold level for pathogens detection. To detect simultaneously the Xcc and LMV in commercial Brassicaceae crop seeds (lettuce, kohlrabi, radish, chinese cabbage and cabbage), two pairs of specific primer (LMV-F/R, Xcc-F/R) were synthesized by using primer-blast program (http://www.ncbi.nlm.nih.gov/tools/primer-blast/). The multiplex PCR for the two pathogens in Brassicaceae crop seeds could detect specifically without interference among primers and/or cDNA of other plant pathogens. The pathogen detection limit was determined at 1 ng of RNA extracted from pathogens. In the total PCR results for pathogen detection using commercial kohlrabi (10 varieties), lettuce (50 varieties), radish (20 varieties), chinese cabbage (20 varieties) and cabbage (20 varieties), LMV and Xcc were detected from 39 and 2 varieties, respectively. In the PCR result of lettuce, LMV and Xcc were simultaneously detected in 8 varieties.

Incidence of Watermelon Mosaic Virus in Cucurbits (박과 작물에 발생하는 Watermelon Mosaic Virus에 관한 연구)

  • Lee Soon Hyung;Lee Key Woon
    • Korean journal of applied entomology
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    • v.20 no.4 s.49
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    • pp.191-195
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    • 1981
  • Cucurbits including pumpkin (Cucurbita pepo), gourd (Lagenariaa siceraria), cucumber (Cucumis sativus), melon(Cucumis melo) and watermelon(Cucurbita anguria) were diseased with mosaic symptoms. The causal virus was identified as watermelon mosaic virus(WMV). The WMV was transmitted by Myzus persicae Sulzer, and no seed borne virus was found. The virus caused large local lesions on the inoculated leaves of the Chenopodium amaranticolor and mosaic symptom on the upper leaves of Cucumis melo, Cucumis sativus, Lagenaria siceraria, Cucurbita anguria and Cucurbita pepo. There were no symptoms on the inoculated leaves of the Nicotiana tabacum var. Bright yellow, Nicotiana glutinosa, Vigna unguiculata. Petunia hybrida and Datura stramonium. Thermal inactivation point was $55\~65^{\circ}C$, dilution end point was $10^{-4}\;10^{-5}$ and longevity in vitro of the virus was $7\~8$ days. The virus showed positive reaction against watermelon mosaic virus antiserum in microprecipitin tests. The virus particles were flexuous rods in size of 750 nm.

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Aphid Over-wintering Host Plants and Seasonal Transmission Rates of Potato Leafroll Virus by Aphids in the Highland Fields of Korea (고랭지 감자밭의 진딧물 월동기주 및 감자잎말림바이러스(PLRV) 보독진딧물의 시기별 변동)

  • Kwon, Min;Kim, Juil;Kim, Changseok;Lee, Yeonggyu
    • Korean journal of applied entomology
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    • v.57 no.4
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    • pp.415-423
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    • 2018
  • Aphid is a typical vector that transfers various kinds of viruses to potatoes. Therefore, it is very important to control aphids moving into potato fields. We investigated the seasonal movement pattern of aphids and its virus transmission rates mainly in the three seed potato production regions at highland in Gangwon-do, Korea. In addition, we identified the aphid species with over-wintering eggs collected from barks or twigs of total 57 tree species around potato fields in winter season. The peak time of summer and winter migration of aphid was at the mid-June and the early October, respectively. A 2.8% of total aphid trapped in yellow water-pan trap was turned out PLRV-borne, and the virus transmission rate was 15.4% by Myzus persicae and 9.1% by Macrosiphum euphorbiae. PLRV-borne aphids started to flow in from the late May, and virus transmission rate of aphid trapped in mid-June was the highest with 10.4%. Totally 14 species of aphid eggs wintered in the 17 species of trees including Acer pictum subsp. mono and Acer pseudosieboldianum at the 11 sites. In particular, because it is not certain that Betula platyphylla var. japonica and Yamatocallis hirayamae do transmit potato virus, but they over-wintered in host plants distributed over a wide area, further research on transmission ability is necessary.

Identification of Bean Common Mosaic Virus Obtained from Seeds of Phaseolus vulgaris (강낭콩에서 종자전염된 Bean Common Mosaic Virus의 분류동정에 관한 연구)

  • Choi Y.M.;Lee S.H.;Park J.S.;Kim J.S.
    • Korean journal of applied entomology
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    • v.23 no.1 s.58
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    • pp.15-21
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    • 1984
  • The virus infecting French bean (Phaseolus vulgaris L.) was identified as Bean Common Mosaic Virus(BCMV) based on the host range, symptomatology, serology, morphology of virus particles and inclusion bodies. Isolates of BCMV were obtained from seeds of P. vulgaris collected at Suweon, Jangsu and Jinju in Korea. French bean produced vein clearing, mosaic, stunting and leaf curling. Symptom of Chenopodium quinoa was local lesions on the inoculated leaves, not on the upper leaves. The electron micrograph of the virus from French bean was flexuous approximately 750nm in length. Cylindrical and pinwheel cytoplasmic inclusion bodies were observed in French bean leaf infected by BCMV. BCMV from the French bean was transmitted through seed and green peach aphid, Myzus persicae. The thermal inactivation point was $55\~60^{\circ}C$, dilution end point was $10^{-3}\~10^{-5}$ and longevity in vitro was $2\~3$ days for BCMV from French bean. The isolates of BCMV reacted positively against BCMV antiserum. The extract of BCMV infected bean leaves, Azukibean mosaic virus (AZMV) and Cowpea aphid borne mosaic virus(CaMV) also reacted with BCMV antiserum, however, BCMV and CaMV showed the spur in agar gel diffusion test.

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Detection of Soybean mosaic virus by Reverse Transcription Loop-mediated Isothermal Amplification (Reverse transcription Loop-mediated isothermal amplification을 이용한 Soybean mosaic virus의 진단)

  • Lee, Yeong-Hoon;Bae, Dae-Hyeon;Kim, Bong-Sub;Yoon, Young-Nam;Bae, Soon-Do;Kim, Hyun-Joo;Mainali, Bishwo P.;Park, In-Hee;Lee, Su-Heon;Kang, Hang-Won
    • Research in Plant Disease
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    • v.21 no.4
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    • pp.315-320
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    • 2015
  • Soybean mosaic virus (SMV) is a prevalent pathogen that causes significant yield reduction in soybean production worldwide. SMV belongs to potyvirus and causes typical symptoms such as mild mosaic, mosaic and necrosis. SMV is seed-borne and also transmitted by aphid. Eleven SMV strains, G1 to G7, G5H, G6H, G7H, and G7a were reported in soybean varieties in Korea. A reverse transcription loop-mediated isothermal amplification (RT-LAMP) method allowed one-step detection of gene amplification by simple procedure and needed only a simple incubator for isothermal template. This RT-LAMP method allowed direct detection of RNA from virus-infected plants without thermal cycling and gel electrophoresis. In this study, we designed RT-LAMP primers named SML-F3/B3/FIP/BIP from coat protein gene sequence of SMV. After the reaction of RT-LAMP, products were identified by electrophoresis and with the detective fluorescent dye, SYBR Green I under daylight and UV light. Optimal reaction condition was at $58^{\circ}C$ for 60 min and the primers of RT-LAMP showed the specificity for nine SMV strains tested in this study.

Bacteriophages: A New Weapon for the Control of Bacterial Blight Disease in Rice Caused by Xanthomonas oryzae

  • Ranjani, Pandurangan;Gowthami, Yaram;Gnanamanickam, Samuel S;Palani, Perumal
    • Microbiology and Biotechnology Letters
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    • v.46 no.4
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    • pp.346-359
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    • 2018
  • Xanthomonas oryzae, a bacterial pathogen causing leaf blight disease (BLB) in rice, can cause widespread disease and has caused epidemics globally, resulting in severe crop losses of 50% in Asia. The pathogen is seed-borne and is transmitted through seeds. Thus, control of BLB requires the elimination of the pathogen from seeds. Concern about environment-friendly organic production has spurred improvements in a variety of biological disease control methods, including the use of bacteriophages, against bacterial plant pathogens. The present study explored the potential of bacteriophages isolated from diseased plant leaves and soil samples in killing the bacterial pathogen in rice seeds. Eight different phages were isolated and evaluated for their bacteriolytic activity against different pathogenic X. oryzae strains. Of these, a phage designated ${\varphi}XOF4$ killed all the pathogenic X. oryzae strains and showed the broadest host range. Transmission electron microscopy of ${\varphi}XOF4$ revealed it to be a tailed phage with an icosahedral head. The virus was assigned to the family Siphoviridae, order Caudovirales. Seedlings raised from the seeds treated with $1{\times}10^8pfu/ml$ of ${\varphi}XOF4$ phage displayed reduced incidence of BLB disease and complete bacterial growth inhibition. The findings indicate the potential of the ${\varphi}XOF4$ phage as a potential biological control agent against BLB disease in rice.