• The Plant Pathology Journal
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• v.18 no.5
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• pp.244-250
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• 2002

Grapevine leafroll-associated 3 virus (GLRaV-3) and Grapevine fanleaf virus (GFLV) are important viral diseases of grapevine in the world. In this study, the most reliable woody indicator plants were selected for virus indexing. Two grapevines, LN33 (Couderc 1613x vitis berlandieri) and Vitis riparia Gloire, were selected for CLRaV-3 and CFLV graft indexing, respectively. The specific characteristics of Closterovirus isolated from grapevines cultivated in Korea were identified. filamentous virus-like particles only existed in the phloem parenchyma cell. In particular, the vesiculation of mitochondria was observed. This mitochondrial vesicu-lation was considered to be one of the most reliable cytopathic features of Closterovirus. During observation of GFLV-infected Chenopodium quinoa sections, virus-like particles arranged consistently were found forming several layers in cytoplasm. Moreover, virus-like particles in tubules were observed and were associated with plasmodesmata in cytoplasm. This is the first report on cytopathological characteristics of Closterovirus and Nepovirus identified from grapevines in Korea.

• The Plant Pathology Journal
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• v.32 no.5
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• pp.452-459
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• 2016

The genomic region of Grapevine fanleaf virus (GFLV) encoding the movement protein (MP) was cloned into pET21a and transformed into Escherichia coli strain BL21 (DE3) to express the protein. Induction was made with a wide range of isopropyl-${\beta}$-D-thiogalactopyranoside (IPTG) concentrations (1, 1.5, and 2 mM) each for duration of 4, 6, or 16 h. However, the highest expression level was achieved with 1 mM IPTG for 4 h. Identity of the expressed protein was confirmed by sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) followed by Western blotting. The expressed 41 kDa protein was purified under denaturing condition by affinity chromatography, reconfirmed by Western blotting and plate-trapped antigen enzyme-linked immunosorbent assay (PTA-ELISA) before being used as a recombinant antigen to raise polyclonal antibodies in rabbits. Purified anti-GFLV MP immunoglobulines (IgGs) and conjugated IgGs detected the expressed MP and GFLV virions in infected grapevines when used in PTA-ELISA, double antibody sandwich-ELISA, and Western blotting. This is the first report on the production of anti-GFLV MP polyclonal antibodies and application for the virus detection.

• Research in Plant Disease
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• v.19 no.3
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• pp.220-225
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• 2013

For quarantine purpose, we developed the RT- and nested PCR module of Tomato black ring virus (TBRV), Arabis mosaic virus (ArMV), Cherry leafroll virus (CLRV) and Grapevine fanleaf virus (GFLV). The PCR modules, developed in this study make diagnosis more convenient and speedy because of same PCR condition. And also, the methods are more accurate because it can check whether the result is contamination or not using the mutation-positive control. We discard or return the 27 cases of Nepovirus infection seed by employing the module past 3 years. This study provides a rapid and useful method for detection of four quarantine plant viruses.