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Effective Application of DAS-ELISA for Detection of Grapevine Leafroll Associated Closterovirus-3 Using a Polyclonal Antiserum Developed from Recombinant Coat Protein

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Abstract

A polyclonal antiserum (As163) specific to grapevine leafroll associated closterovirus-3 (GLRaV-3) was developed using a recombinant coat protein expressed in E. coli from a cDNA clone identified after immunoscreening of a cDNA library. Specificity of the antiserum to GLRaV-3 was shown by Western blot and immunosorbent electron microscopy. With this antiserum, an effective double antibody sandwich enzyme-linked immunosorbent assay (DAS-ELISA) was developed for GLRaV-3 detection. To evaluate the sensitivity of the antiserum in DAS-ELISA for virus detection, different combinations of antibodies were compared. Although best results were obtained when As163 was used for coating and a monoclonal antibody (MabNY1.1) was used as an enzyme conjugate, good results were also obtained when As163 was used both for coating and as an enzyme conjugate. Using this As163–Mab system in DAS-ELISA, we confirmed the presence of GLRaV-3 in a diverse collection of leafroll infected vines.

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References

  • Agran MK, Di Terlizzi B, Boscia D, Minafra A, Savino V, Martelli GP and Askri F (1990) Occurrence of grapevine virus A (GVA) and other closteroviruses in Tunisian grapevines affected by leafroll disease. Vitis 29: 43–48

    Google Scholar 

  • Belli G, Fortusini A, Casati P, Belli L, Bianco PA and Prati S (1994) Transmission of a grapevine leafroll associated closterovirus by the scale insect Pulvinaria vitis L. Rivista di Patologia Vegetale 4: 105–108

    Google Scholar 

  • Boscia D, Greif C, Gugerli P, Martelli GP, Walter B and Gonsalves D (1995) Nomenclature of grapevine leafrollassociated putative closteroviruses. Vitis 34: 171–175

    Google Scholar 

  • Choueiri E, Boscia D, Digiaro M, Castellano MA and Martelli GP (1996) Some properties of a hitherto undescribed filamentous virus of the grapevine. Vitis 35: 1–3

    Google Scholar 

  • Clark MF and Adams AN (1977) Characteristics of the microplate method for enzyme-linked immunosorbent assay for the detection of plant viruses. J Gen Virol 34: 475–483

    PubMed  Google Scholar 

  • Credi R and Giunchedi L (1996) Grapevine leafroll-associated viruses and grapevine virusAin selected Vitis vinifera cultivars in northern Italy. Plant Pathology 45: 1110–1116

    Google Scholar 

  • Engelbrecht DJ and Kasdorf GGF (1990) Transmission of grapevine leafroll diseases and associated closteroviruses by the vine mealybug, Planococcus ficus. Phytophylactica 22: 341–346

    Google Scholar 

  • Englard S and Seifter S (1990) Precipitation techniques. Method Enzymol 182: 285–300

    Google Scholar 

  • Forsline PL, Hoch J, Lamboy WF, Hu JS, McFerson D, Golino D and Gonsalves D (1996) Comparative effectiveness of symptomatology and ELISA for detecting two isolates of grapevine leafroll on graft-inoculated Cabernet Franc. American J Enol Vitic 47: 239–243

    Google Scholar 

  • Golino DA (1992) The Davis grapevine virus collection. American J Enol Vitic 43: 200–205

    Google Scholar 

  • Golino DA, Sim ST, Gill R and Rowhani A (1998) Transmission studies of grapevine closteroviruses by four species of mealybugs. Phytopathology 88: S32.

    Google Scholar 

  • Golino DA, Sim ST and Rowhani A (1995) Transmission studies of grapevine leafroll associated virus and grapevine corky bark associated virus by the obscure mealybug. American J Enol Vitic 46: 408.

    Google Scholar 

  • Goszczynski DE, Kasdorf GGF, Pietersen G and Van Tonder H (1996) Grapevine leafroll-associated virus-2 (GLRaV-2)-mechanical transmission, purification, production and properties of antisera, detection by ELISA. S African J Enol Vitic 17: 15–26

    Google Scholar 

  • Hager DA and Burgess RR (1980) Elution of proteins from sodium dodecyl sulfate-polyacrylamide gels, removal of sodium dodecyl sulfate, and renaturation of enzymatic activity: results with sigma subunit of E. coli RNA polymerase, wheat germ DNA topoisomerase and other enzymes. Anal Biochem 109: 76–86

    PubMed  Google Scholar 

  • Hu JS, Gonsalves D, Boscia D and Namba S (1990a) Use of monoclonal antibodies to characterize grapevine leafroll associated closteroviruses. Phytopathology 80: 920–925

    Google Scholar 

  • Hu JS, Gonsalves D and Teliz D (1990b) Characterization of closterovirus-like particles associated with grapevine leafroll disease. J Phytopathol 128: 1–14

    Google Scholar 

  • Jelkmann W and Keim-Konrad R (1997) Immuno-capture polymerase chain reaction and plate-trapped ELISA for the detection of apple stem pitting virus. J Phytopathol 145: 499–503

    Google Scholar 

  • Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227: 680–685

    PubMed  Google Scholar 

  • Ling KS, Provvidenti R and Gonsalves D (1995) Detection of turnip mosaic virus isolates using an antiserum to coat protein breakdown products. Plant Dis 79: 809–812

    Google Scholar 

  • Ling KS, Zhu HY, Alvizo H, Hu JS, Drong RF, Slightom JL and Gonsalves D (1997) The coat protein gene of grapevine leafroll associated closterovirus-3: cloning, nucleotide sequencing and expression in transgenic plants. Arch Virol 142: 1101–1116

    PubMed  Google Scholar 

  • Ling KS, Zhu HY, Drong RF, Slightom JL and Gonsalves D (1998) Nucleotide sequence of the 30 terminal two-thirds of the grapevine leafroll-associated virus-3 genome reveals a typical monopartite closterovirus. J Gen Virol 79: 1299–1307

    PubMed  Google Scholar 

  • Milne R and Luisoni E (1977) Rapid immune electron microscopy of virus preparations. Meth Virol 6: 265–281

    Google Scholar 

  • Monis J and Bestwick RK (1997) Serological detection of grapevine associated closteroviruses in infected grapevine cultivars. Plant Dis 81: 802–808

    Google Scholar 

  • Namba S, Boscia D, Azzam O, Maixner M, Hu JS, Golino D and Gonsalves D (1991) Purification and properties of closterovirus-like particles associated with grapevine corky bark disease. Phytopathology 81: 964–970

    Google Scholar 

  • Nikolaeva OV, Karasev AV, Gumpf DJ, Lee RF and Garnsey SM (1995) Production of polyclonal antisera to the coat protein of citrus tristeza virus expressed in Escherichia coli: application for immunodiagnosis. Phytopathology 85: 691–694

    Google Scholar 

  • Petersen CL and Charles JG (1997) Transmission of grapevine leafroll-associated closteroviruses by Pseudococcus longispinus and P. calceolariae. Plant Pathol 46: 509–515

    Google Scholar 

  • Purcifull DE and Batchelor DL (1977) Immunodiffusion tests with sodium dodecyl sulfate (SDS) treated plant viruses and plant virus inclusions. Fla Agric Exp Stn Bull 788

  • Rosciglione B and Gugerli P (1989) Transmission of grapevine leafroll disease and an associated closterovirus to healthy grapevine by the mealybug Planococcus ficus (Abstract). Phytoparasitica 17: 63

    Google Scholar 

  • Rubinson E, Galiakparov N, Radian S, Sela I, Tanne E and Gafny R (1997) Serological detection of grapevine virus A using antiserum to a nonstructural protein, the putative movement protein. Phytopathology 87: 1041–1045

    Google Scholar 

  • Tanne E, Ben DY and Raccah B (1989) Transmission of closterovirus-like particles by mealybugs (Abstract). Phytoparasitica 17: 63–64

    Google Scholar 

  • Vaira AM, Vecchiati M, Masenga V and Accotto GP (1996) A polyclonal antiserum against a recombinant viral protein combines specificity with versatility. J Virol Meth 56: 209–219

    Google Scholar 

  • Walter B and Etienne L (1987) Detection of the grapevine fanleaf viruses away from the period of vegetation. J Phytopathol 120: 355–364

    Google Scholar 

  • Zee F, Gonsalves D, Goheen A, Kim KS, Pool R and Lee RF (1987) Cytopathology of leafroll-diseased grapevines and the purification and serology of associated closteroviruslike particles. Phytopathology 77: 1427–1434

    Google Scholar 

  • Zhu HY, Ling KS, Goszcyski DE, McFerson J and Gonsalves D (1998) Nucleotide sequence and genome organization of grapevine leafroll associated closterovirus-2 is similar to beet yellows virus, the Closteovirus type member. J Gen Virol 79: 1289–1298

    PubMed  Google Scholar 

  • Zimmermann D, Bass P, Legin R and Walter B (1990a) Characterization and serological detection of four closterovirus-like particles associated with leafroll disease on grapevine. J Phytopathol 130: 205–218

    Google Scholar 

  • Zimmermann D, Sommermeyer G, Walter B and Van RMHV (1990b) Production and characterization of monoclonal antibodies specific to closterovirus-like particles associated with grapevine leafroll disease. J Phytopathol 130: 277–288

    Google Scholar 

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Author notes

  1. Kai-Shu Ling

    Present address: AgriVitis, Inc., 2191 San Juan-Hollister Road, San Juan Bautista, CA, 95045, USA

Authors and Affiliations

  1. Department of Plant Pathology, NYS Agricultural Experimental Station, Cornell University, Geneva, NY, 14456, USA

    Kai-Shu Ling, Hai-Ying Zhu & Zhao-Yuan Jiang

  2. Department of Plant Pathology, NYS Agricultural Experimental Station, Cornell University, Geneva, NY, 14456, USA;

    Dennis Gonsalves

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  1. Kai-Shu Ling
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  2. Hai-Ying Zhu
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  3. Zhao-Yuan Jiang
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  4. Dennis Gonsalves
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Ling, KS., Zhu, HY., Jiang, ZY. et al. Effective Application of DAS-ELISA for Detection of Grapevine Leafroll Associated Closterovirus-3 Using a Polyclonal Antiserum Developed from Recombinant Coat Protein. European Journal of Plant Pathology 106, 301–309 (2000). https://doi.org/10.1023/A:1008713319294

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