Use of Cross‐Reactive Antibodies to Detect Members of the <i>Potyviridae</i>

Richter, Johannes; Rabenstein, Frank; Proll, E.; Vetten, H. J.

doi:10.1111/j.1439-0434.1995.tb04554.x

Cited by 20 publications

(9 citation statements)

References 30 publications

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“…To rule out the possibility that an aphid-borne potyvirus was contaminating the CVYV cultures, three series of experiments were conducted. CVYV-infected cucumber and melon plant extracts were tested in double-antibody sandwich (DAS)-ELISA with antisera raised against the five most common potyviruses encountered in cucurbits in the Mediterranean region (Lecoq et al, 1998) In addition, CVYV-infected plant extracts did not react in antigen-trapped indirect ELISA with monoclonal antibody PTY 1, which detects most aphid-transmitted potyviruses but which does not detect potyvirids transmitted by other vectors with the exception of Ryegrass mosaic virus (RGMV) (Jordan, 1992 ;Richter et al, 1995) (data not shown). Finally, attempts to transmit CVYV-Isr and CVYV-Jor by the aphid Myzus persicae by the non-persistent mode were unsuccessful (data not shown).…”

Section: Cucumber Vein Yellowing Virus (Cvyv) Is Widespread In Cucurbmentioning

confidence: 99%

Cytological and molecular evidence that the whitefly-transmitted Cucumber vein yellowing virus is a tentative member of the family Potyviridae

Lecoq¹,

Desbiez²,

Delécolle³

et al. 2000

Journal of General Virology

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Cucumber vein yellowing virus (CVYV) is widespread in cucurbits in the Middle East. CVYV has filamentous particles and is transmitted byBemisia tabaci by the semi-persistent mode. It has not yet been assigned to a specific genus or family. Ultramicroscopic observations revealed numerous cylindrical cytoplasmic inclusions in melon and cucumber cells infected by CVYV isolates from Israel and Jordan. Depending on the section orientation, the inclusions appeared as pinwheels or as bundles. In addition, a 1n9 kb DNA fragment was amplified by RT-PCR from CVYV-infected plant extracts using primers designed to detect all potyvirids. Sequence comparisons with the amplified fragment indicated that CVYV is more closely related to Sweet potato mild mottle virus than to any other virus in the family Potyviridae. These results suggest that CVYV can be considered as a tentative new member of the genus Ipomovirus, family Potyviridae.Cucumber vein yellowing virus (CVYV) causes a severe disease of cucumbers and other cucurbits in the Oriental Mediterranean Basin (Cohen & Nitzany, 1960 ;Harpaz & Cohen, 1965 ;Al-Musa et al., 1985 ;Yilmaz et al., 1989). CVYV is reported to have rod-shaped particles, 740-800 nm long with a diameter of 15-18 nm (Sela et al., 1980). The virus is readily transmitted mechanically and by the whitefly Bemisia tabaci by the semi-persistent mode (Harpaz & Cohen, 1965 ;Mansour & Al-Musa, 1993). Due to the instability of CVYV particles, difficulties have been encountered in obtaining purified virus preparations (Sela et al., 1980 ;Mansour & Hadidi, 1999) ; therefore, very little is known of the biophysical Author for correspondence : Herve! Lecoq.Fax j33 4 32 72 28 42. e-mail Herve.Lecoq!avignon.inra.frThe GenBank accession number of the sequence reported in this paper is AF233429. and biochemical properties of CVYV and the virus is still unclassified (Brunt et al., 1996 ;Lecoq et al., 1998). However, Sela et al. (1980) have reported that the CVYV coat protein (CP) has a molecular mass of 39 kDa and that the viral nucleic acid is a double-stranded DNA, suggesting that CVYV could be a member of a new virus family.In order to investigate the taxonomic position of CVYV further, cytopathological, serological and molecular studies were conducted. Cucumber (Cucumis sativus cv. Beit Alfa) and melon (Cucumis melo cv. Ve! drantais) plants were inoculated mechanically with the CVYV type strain from Israel (CVYVIsr) or with a CVYV isolate from Jordan (CVYV-Jor) and were kept subsequently in independent screened cages maintained in an insect-proof greenhouse. The two strains induced similar vein-clearing symptoms in cucumber and melon, but CVYVJor caused a more severe stunting in cucumber.For cytological studies, leaf pieces 1 mm across were collected from symptomatic young leaves 3-4 weeks after inoculation and similar samples from healthy plants were used as controls. Samples were fixed with glutaraldehyde, post-fixed with osmium tetroxide and embedded in araldite CY212 (Agar Scientific) (Dele! colle, 1978). Thin sectio...

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Section: Cucumber Vein Yellowing Virus (Cvyv) Is Widespread In Cucurbmentioning

confidence: 99%

Cytological and molecular evidence that the whitefly-transmitted Cucumber vein yellowing virus is a tentative member of the family Potyviridae

Lecoq¹,

Desbiez²,

Delécolle³

et al. 2000

Journal of General Virology

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“…data) and recently published 3' sequence information from a Danish isolate of RgMV, suggest that mite-transmitted RgMV is more closely related to viruses of the aphid-transmitted potyvirus genus (Schubert et al 1995). This assertion is supported by the serological reaction of RgMV to a number of general potyvirus antisera including PTY1 (Richter et al 1995). PTY1 reacts with mostpotyviruses, but with the exception of RgMV, does not react with rymo-, bymo-, or ipomoviruses (Richter et al 1995).…”

Section: Introductionmentioning

confidence: 88%

“…This assertion is supported by the serological reaction of RgMV to a number of general potyvirus antisera including PTY1 (Richter et al 1995). PTY1 reacts with mostpotyviruses, but with the exception of RgMV, does not react with rymo-, bymo-, or ipomoviruses (Richter et al 1995).…”

Section: Introductionmentioning

confidence: 91%

Distribution of ryegrass mosaic virus in New Zealand perennial ryegrass pastures as determined by ELISA and RT‐PCR

Webster

Forster

Sinclair

et al. 1996

New Zealand Journal of Agricultural Research

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Ryegrass mosaic virus (RgMV) is a mite-transmitted member of the Potyviridae that can seriously affect yield and persistence of ryegrass in pastures. To determine the incidence and distribution of RgMV in New Zealand, perennial ryegrass pastures were surveyed by indirect ELISA. RgMV was detected in all seven mature ( 4 years) North Island pastures and 60% of 20 mature South Island pastures, indicating that RgMV is established throughout New Zealand. A reverse transcriptasepolymerase chain reaction (RT-PCR) assay was developed to confirm the identity of RgMV. This assay was 200 times more sensitive than the indirect ELISA. A possible correlation between numbers of the eriophyid mites and the incidence of RgMV was observed in South Island pastures. Our results suggest that mite transmission may be more significant than ryegrass cultivar, pasture age, endophyte infection, or mechanical transmission in defining the incidence and distribution of RgMV. Options for the control of RgMV in New Zealand pastures are discussed.

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“…First, we attempted to purify virus particles from infected plants for use as immunogens and we then screened for MAbs that recognized several potyviruses, similar to previous reports of MAbs prepared against Johnsongrass mosaic virus (JGMV) (42), Tulip breaking virus (43), TEV (44,45), and Peanut stripe virus (PStV) (46). Unfortunately, it was difficult to purify enough virus particles from calla lily to produce high-quality antiserum or antibody for detection, most likely due to the mucilage in the plant tissues (17,47).…”

Section: Discussionmentioning

confidence: 99%

A Strategy for Generating a Broad-Spectrum Monoclonal Antibody and Soluble Single-Chain Variable Fragments against Plant Potyviruses

Liu

Lin

et al. 2015

Appl Environ Microbiol

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Potyviruses are major pathogens that often cause mixed infection in calla lilies. To reduce the time and cost of virus indexing, a detection method for the simultaneous targeting of multiple potyviruses was developed by generating a broad-spectrum monoclonal antibody (MAb) for detecting the greatest possible number of potyviruses. The conserved 121-amino-acid core regions of the capsid proteins of Dasheen mosaic potyvirus (DsMV), Konjak mosaic potyvirus (KoMV), and Zantedeschia mild mosaic potyvirus (ZaMMV) were sequentially concatenated and expressed as a recombinant protein for immunization. After hybridoma cell fusion and selection, one stable cell line that secreted a group-specific antibody, named C4 MAb, was selected. In the reaction spectrum test, the C4 MAb detected at least 14 potyviruses by indirect enzyme-linked immunosorbent assay (I-ELISA) and Western blot analysis. Furthermore, the variable regions of the heavy (V H ) and light (V L ) chains of the C4 MAb were separately cloned and constructed as single-chain variable fragments (scFvs) for expression in Escherichia coli. Moreover, the pectate lyase E (PelE) signal peptide of Erwinia chrysanthemi S3-1 was added to promote the secretion of C4 scFvs into the medium. According to Western blot analysis and I-ELISA, the soluble C4 scFv (V L -V H ) fragment showed a binding specificity similar to that of the C4 MAb. Our results demonstrate that a recombinant protein derived from fusion of the conserved regions of viral proteins has the potential to produce a broad-spectrum MAb against a large group of viruses and that the PelE signal peptide can improve the secretion of scFvs in E. coli. The genus Potyvirus, one of the largest groups of plant viruses, includes 158 species, many of which are economically important, such as Potato virus Y (PVY) and Plum pox virus (1, 2). Most potyviruses infect various host plants and are spread worldwide by aphid vectors and mechanical inoculation (3). Potyviruses have a single-stranded, positive-sense RNA genome of approximately 10 kb in length that encodes a polyprotein translated from a large open reading frame (ORF) (3) and a trans-frame movement-related protein (P3N-PIPO) encoded from the 5=-terminal portion of P3 and the PIPO cistrons (4, 5). During proteolytic processing, the large polyprotein can be separated into 10 mature proteins that are involved in virus replication, movement, and virion assembly (3). Potyviral capsid proteins (CPs) not only form the viral particle but also participate in cell-to-cell and systemic movement (6, 7). The variable N-and C-terminal regions of the CPs exposed on the virion surface are required for long-distance transport, are involved in aphid transmission (DAG motif), and may also assist in cell-to-cell spreading (7-10), whereas the conserved core regions of CPs are necessary for virion assembly and the cell-to-cell movement of potyviruses (6, 7).In Taiwan, the viral disease caused by potyviruses is one of the limiting factors for production of calla lilies. At least five potyvirus...

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Use of Cross‐Reactive Antibodies to Detect Members of the Potyviridae

Cited by 20 publications

References 30 publications

Cytological and molecular evidence that the whitefly-transmitted Cucumber vein yellowing virus is a tentative member of the family Potyviridae

Cytological and molecular evidence that the whitefly-transmitted Cucumber vein yellowing virus is a tentative member of the family Potyviridae

Distribution of ryegrass mosaic virus in New Zealand perennial ryegrass pastures as determined by ELISA and RT‐PCR

A Strategy for Generating a Broad-Spectrum Monoclonal Antibody and Soluble Single-Chain Variable Fragments against Plant Potyviruses

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