Abstract
Eight isolates of Grapevine virus A (GVA), which induced different symptoms in leaves of Nicotiana benthamiana, were recovered from various grapevines. The dsRNA patterns of two isolates, which consistently induced mild vein clearing (referred here as mild isolates of GVA) were similar, but different from those of other isolates of GVA. Analysis based on overall nucleotide (nt) sequence identity in the 3′ terminal part of the GVA genome, comprising part of ORF3 (putative movement protein, MP), entire ORF4 (capsid protein, CP), entire ORF5 and part of 3′ UTR, revealed that GVA isolates separate into three groups (I, II, III), sharing 91.0–99.8% nt sequence identity within groups and 78.0–89.3% nt sequence identity between groups. Mild isolates of the virus were group III and shared only 78.0–79.6% nt sequence identity with the other isolates. The comparison of predicted amino acid sequences for MP and CP revealed many amino acid alterations, revealing distinct local net charges of these proteins for mild isolates of the virus. Based on both conserved and divergent nt regions in the CP and ORF5, oligonucleotide primers were designed for the simultaneous RT-PCR detection of all GVA isolates and for the specific detection of the most divergent virus variants represented here by mild isolates of the virus.
Similar content being viewed by others
References
Ayllon MA, Lopez C, Navas-Castillo J, Garnsey SM, Guerri J, Flores R and Moreno P (2001) Polymorphism of the 5' terminal region of Citrus tristeza virus (CTV) RNA: Incidence of three sequence types in isolates of different origin and pathogenicity. Archives of Virology 146: 27-40
Banerjee N, Wang J-Y and Zaitlin M (1995) A single nucleotide change in the coat protein gene of tobacco mosaic virus is involved in the induction of severe chlorosis. Virology 207: 234-239
Bol JF (1999) Alfalfa mosaic virus and ilarviruses: Involvement of coat protein in multiple steps of the replication cycle. Journal of General Virology 80: 1089-1102
Boscia D, Minafra A and Martelli GP (1997) Filamentous viruses of grapevine: Putative trichoviruses and capilloviruses. In: Monette PL (ed) Filamentous Viruses of Woody Plants (pp 19-28) Research Signpost, Trivandrum, India
Chevalier S, Grief C, Clauzel JM, Walter B and Fritsch C (1995) Use of immunocapture-polymerase chain reaction for the detection of grapevine virus A in kober stem grooving-infected grapevines. Journal of Phytopathology 143: 369-373
Choueiri E, Digiaro M and Savino V (1997) Further evidence that grapevine virusAis the agent of kober stem grooving. Extended abstracts 12th Meeting ICVG, Lisbon, Portugal, 29 September-2 October (pp 39-40)
Conti M, Milne RG, Luisoni E and Boccardo C (1980) A closterovirus from a stem-pitting affected grapevine. Phytopathology 70: 394-399
De Meyer J, Gaudin M, Bourquin L, Jakab G, Malnoe P and Gugerli P (2000) New primers for the molecular identification and detection of grapevine virus A (GVA). In: Extended abstracts 13th Meeting ICVG, Adelaide, 12-17 March (pp 138-140)
Garau R, Prota VA, Piredda R, Boscia D and Prota U (1994) On the possible relationship between Kober stem grooving and grapevine virus A. Vitis 33: 161-163
Goszczynski DE, Kasdorf GGF and Pietersen G (1996) Western blot reveal that grapevine virus A and grapevine virus B are serologically related. Journal of Phytopathology 144: 581-583
Goszczynski DE and Jooste AEC (2002) The application of single-strand conformation polymorphism (SSCP) technique for the analysis of molecular heterogeneity of grapevine virus A. Vitis 41: 77-82
Hu JS, Gonsalves D and Teliz D (1990) Characterization of closterovirus-like particles associated with grapevine leafroll disease. Journal of Phytopathology 128: 133-135
Martelli GP, Minafra A and Sardarelli P (1997) Vitivirus, a new genus of plant viruses. Archives of Virology 142: 1929-1932
Minafra A, Saldarelli P and Martelli GP (1997) Grapevine virus A: Nucleotide sequence, genome organization, and relationship in the Trichovirus genus. Archives of Virology 142: 417-423
Minafra A (2000) Rugose wood of grapevines. In: Extended abstracts 13th Meeting ICVG, Adelaide, Australia, 12-17 March (pp 30-34)
Monette PL, Godkin SE and James D (1990) Mechanical sap transmission of a closterovirus from in vitro shoot tip cultures of a leafroll-affected grapevine to Nicotiana benthamiana. Vitis 29: 49-55
Moreno P, Guerri J, Ballester-Olmos JF, Albiach R and Martinez ME (1993) Separation and interference of strains from a citrus tristeza virus isolate evidenced by biological activity and double-stranded RNA (dsRNA) analysis. Plant Pathology 42: 35-41
Moreno IM, Bernal JJ, de Blas BG, Rodriguez-Cerezo E and Garcia-Arenal F (1997) The expression level of the 3a movement protein determines differences in severity of symptoms between two strains of tomato aspermy cucumovirus. Molecular Plant-Microbe Interactions 10: 171-179
Valverde RA, Nementh ST and Jordan RL (1990) Analysis of double-stranded RNA for plant virus diagnosis. Plant Disease 74: 255-258
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Goszczynski, D., Jooste, A. Identification of divergent variants of Grapevine virus A. European Journal of Plant Pathology 109, 397–403 (2003). https://doi.org/10.1023/A:1023555018700
Issue Date:
DOI: https://doi.org/10.1023/A:1023555018700