scholarly journals First Detection of Rupestris stem pitting associated virus Particles by Antibody to a Recombinant Coat Protein

Plant Disease ◽  
2003 ◽  
Vol 87 (5) ◽  
pp. 510-514 ◽  
Author(s):  
Natasa Petrovic ◽  
Baozhong Meng ◽  
Maja Ravnikar ◽  
Irena Mavric ◽  
Dennis Gonsalves
Keyword(s):  
Coat Protein ◽  
Polyclonal Antiserum ◽  
Enzyme Linked Immunosorbent Assay ◽  
Grapevine Virus ◽  
Virus Particles ◽  
Grapevine Virus A ◽  
Grapevine Virus B ◽  
Rupestris Stem Pitting ◽  
Recombinant Coat Protein ◽  
Stem Pitting

Rupestris stem pitting associated virus (RSPaV), a member of the genus Foveavirus, is associated with the Rupestris stem pitting component of the Rugose wood (RW) disease complex of grapevines. Heretofore, particles of RSPaV have not been visualized. In this work, flexuous rod particles approximately 723 nm in length were detected in the sap of infected grapevines by immunosorbent electron microscopy (ISEM), using a polyclonal antiserum produced to a recombinant coat protein of RSPaV. Particles of RSPaV were detected in tissue culture-, greenhouse-, and field-grown grapevines infected with RSPaV, but not in healthy control plants. Detection of virus particles by ISEM corresponded with detection of RSPaV by Western blot, enzyme-linked immunosorbent assay, and reverse transcription-polymerase chain reaction. Virus particles were decorated with the antibodies specific to RSPaV but not with antibodies to Grapevine virus A or Grapevine virus B, two other viruses believed to be associated with RW. This definitive identification of RSPaV particles will help define the etiology of RW.

scholarly journals Incidência de vírus em videiras no Nordeste brasileiro e caracterização molecular parcial de isolados virais locais

2015 ◽  
Vol 45 (3) ◽  
pp. 379-385 ◽  
Author(s):  
Aricléia de Moraes Catarino ◽  
Thor Vinícius Martins Fajardo ◽  
Gilvan Pio-Ribeiro ◽  
Marcelo Eiras ◽  
Osmar Nickel
Keyword(s):  
Grapevine Virus ◽  
Rt Pcr ◽  
Grapevine Fanleaf Virus ◽  
Grapevine Virus A ◽  
Grapevine Virus B ◽  
Rupestris Stem Pitting ◽  
Stem Pitting

Os objetivos deste trabalho foram identificar as espécies virais presentes em vinhedos comerciais de duas regiões do Nordeste do Brasil e realizar a caracterização molecular parcial de isolados de três espécies virais. A diagnose foi realizada por meio de RT-PCR em tempo real para a detecção de Grapevine rupestris stem pitting-associated virus (GRSPaV), Grapevine virus A (GVA), Grapevine virus B (GVB), Grapevine leafroll-associated virus 2, 3 e 4 (GLRaV-2, -3 e -4), Grapevine fleck virus (GFkV), Grapevine rupestris vein feathering virus (GRVFV) e Grapevine fanleaf virus (GFLV). Exceto para GFLV, os vírus avaliados estão amplamente disseminados nas áreas amostradas, frequentemente em altas incidências e em infecções múltiplas, de até 98% e 76,4%, na Zona da Mata e no Vale do São Francisco, respectivamente. Isolados locais de GVA, GVB e GLRaV-3 foram parcialmente caracterizados com base na sequência completa de nucleotídeos do gene da proteína capsidial e apresentaram alta porcentagem de identidade de nucleotídeos com outros isolados brasileiros: 91,2% (GVA), 99,8% (GVB) e 99,7% (GLRaV-3)

scholarly journals Antiserum to Recombinant Virus Coat Protein Detects Rupestris stem pitting associated virus in Grapevines

Plant Disease ◽  
2003 ◽  
Vol 87 (5) ◽  
pp. 515-522 ◽  
Author(s):  
Baozhong Meng ◽  
Rino Credi ◽  
Natasa Petrovic ◽  
Irma Tomazic ◽  
Dennis Gonsalves
Keyword(s):  
Coat Protein ◽  
Virus Disease ◽  
Biological Indicator ◽  
Polyclonal Antiserum ◽  
Enzyme Linked Immunosorbent Assay ◽  
Vitis Rupestris ◽  
Biological Indexing ◽  
Virus Coat ◽  
Rupestris Stem Pitting ◽  
Stem Pitting

Rupestris stem pitting (RSP) is the most widespread virus disease of grapevines. The genome of Rupestris stem pitting associated virus (RSPaV), the putative causal agent of RSP, was recently sequenced. Until recently, the only method to diagnose RSP was biological indexing on woody indicator plants, a process that takes 2 to 3 years to complete. This study reports on the production of a polyclonal antiserum to a recombinant coat protein of RSPaV. The antiserum was used effectively to detect RSPaV from various genotypes and tissues of grapevines by Western blot and indirect enzyme-linked immunosorbent assay. Virus antigens were consistently detected in the cambium of dormant canes and in actively growing leaves of grapevines. Moreover, plants of Vitis rupestris ‘St. George’, the standard biological indicator for RSP, tested positive for RSPaV. The serological methods developed in this study are advantageous as compared with biological indexing because they are more rapid, less expensive, as reliable, and more suitable for assays of a large number of samples.

scholarly journals Desempenho agronômico de videiras com e sem sintomas de viroses, e comparação molecular de isolados virais

2015 ◽  
Vol 50 (7) ◽  
pp. 541-550 ◽  
Author(s):  
Monique Bezerra Nascimento ◽  
Thor Vinícius Martins Fajardo ◽  
Marcelo Eiras ◽  
Ana Beatriz Costa Czermainski ◽  
Osmar Nickel ◽  
...  
Keyword(s):  
Grapevine Virus ◽  
Rt Pcr ◽  
Grapevine Virus A ◽  
Grapevine Virus B ◽  
Rupestris Stem Pitting ◽  
Stem Pitting

Resumo: O objetivo deste trabalho foi avaliar os efeitos de viroses em videiras sintomáticas e assintomáticas sobre as variáveis agronômicas relacionadas ao vigor das plantas e à qualidade enológica da uva, e comparar os isolados virais obtidos nessas duas condições. Realizaram-se dois experimentos com quatro cultivares. Todas as plantas foram indexadas, por meio da reação em cadeia da polimerase via transcrição reversa (RT-PCR) em tempo real, quanto à provável ocorrência dos seguintes vírus: Grapevine virus A (GVA), Grapevine virus B (GVB), Grapevine virus D (GVD), Grapevine leafroll-associated virus (GLRaV-1 ao -4, GLRaV-4 estirpe 5), Grapevine rupestris stem pitting-associated virus (GRSPaV) e Grapevine fleck virus (GFkV). As variáveis avaliadas foram: número de gemas brotadas e não brotadas, número de ramos com ou sem cachos, número total de gemas, número de cachos, massa de cachos frescos, massa total de bagas, massa do engaço, número de bagas por cacho, massa média de baga, sólidos solúveis totais, acidez total titulável, pH, massa de ramos podados ou diâmetros do tronco do porta-enxerto e da copa. Os efeitos negativos foram mais pronunciados nas plantas com sintomas de viroses; no entanto, constatou-se frequentemente que plantas sem sintomas também estavam infectadas. A análise molecular de GRSPaV, GVA e GLRaV-2, isolados de plantas sintomáticas e assintomáticas, resultou em alta percentagem de identidade de nucleotídeos entre isolados homólogos.

scholarly journals A Survey of Virginia Vineyards Revealed High Incidences of Grapevine Rupestris Stem Pitting-Associated Virus, Grapevine Red Blotch Virus, and Two Mealybug Species

2019 ◽  
Vol 20 (4) ◽  
pp. 207-214 ◽  
Author(s):  
Taylor Jones ◽  
Mizuho Nita
Keyword(s):  
Virus Disease ◽  
Grapevine Virus ◽  
Factors Affecting ◽  
Grapevine Virus A ◽  
Grapevine Virus B ◽  
Grapevine Pinot Gris Virus ◽  
Potential Factors ◽  
Pseudococcus Viburni ◽  
Rupestris Stem Pitting ◽  
Stem Pitting

We investigated the prevalence of viruses infecting grapevines in Virginia, identity of disease vectors, and potential factors affecting virus incidence. Tested viruses were grapevine leafroll-associated virus (GLRaV-1 and -4), grapevine fleck virus (GFkV), grapevine virus A (GVA), grapevine virus B (GVB), grapevine rupestris stem pitting-associated virus (GRSPaV), tomato ringspot virus (ToRSV), grapevine vein clearing virus (GVCV), grapevine red blotch virus (GRBV), and grapevine Pinot gris virus (GPGV). We documented wide distributions of GRSPaV (54%) and GRBV (24%) and common occurrences of grape (Pseudococcus maritimus) and Gill’s (Ferrisia gilli) mealybugs among vineyards. This is the first report of GLRaV-1, GLRaV-4, GVA, GVB, GRSPaV, and obscure mealybug (Pseudococcus viburni) in Virginia. We also documented significant association (P ≤ 0.05) of the presence of mealybugs and GVA and GVB. With younger vines, significantly lower incidences were found for viruses that were listed (i.e., tested for a certification) by the Foundation Planting Service’s and the National Clean Plant Network’s grape programs. On the other hand, there was a lack of the age effect on incidence of GRSPaV and GRBV, which were not listed until recently. These results suggest the importance of clean plant material and vector management for grapevine virus disease management in Virginia.

scholarly journals Detecção e identificação molecular de vírus associados a videiras sintomáticas e assintomáticas

2010 ◽  
Vol 40 (11) ◽  
pp. 2249-2255 ◽  
Author(s):  
Marcos Fernando Basso ◽  
Thor Vinícius Martins Fajardo ◽  
Marcelo Eiras ◽  
Ricardo Antônio Ayub ◽  
Osmar Nickel
Keyword(s):  
Mosaic Virus ◽  
Grapevine Virus ◽  
Rt Pcr ◽  
Grapevine Fanleaf Virus ◽  
Arabis Mosaic Virus ◽  
Grapevine Virus A ◽  
Grapevine Virus B ◽  
Para Determinar ◽  
Rupestris Stem Pitting ◽  
Stem Pitting

A propagação vegetativa da videira favorece infecções virais múltiplas, com expressão diferencial de sintomas em função da combinação da cultivar ou espécie da hospedeira com a espécie viral. Os objetivos deste trabalho foram detectar e identificar as espécies virais presentes em duas espécies/cultivares de videira: uma sintomática e outra assintomática. DsRNA de ambas as amostras foi submetido à RT-PCR com 17 pares de oligonucleotídeos específicos para a detecção de Grapevine virus A (GVA), Grapevine virus B (GVB), Grapevine virus D (GVD), Grapevine fleck virus (GFkV), Grapevine fanleaf virus (GFLV), Arabis mosaic virus (ArMV), Grapevine chrome mosaic virus (GCMV), Rupestris stem pitting-associated virus (RSPaV), Grapevine leafroll-associated virus 1 a 4 (GLRaV-1 a -4), além de três pares de oligonucleotídeos degenerados. Pelo menos um fragmento amplificado, por par de oligonucleotídeos, foi clonado e sequenciado. Plantas sintomáticas e assintomáticas mostraram infecções múltiplas por RSPaV, GLRaV-2 e/ou GLRaV-3. As sequências de nucleotídeos obtidas para sete isolados de RSPaV, três de GLRaV-2 e dois de GLRaV-3 apresentaram identidades superiores a 90% com espécies homólogas e permitiram a definição das possíveis estirpes presentes nas amostras infectadas. Esses resultados evidenciam a necessidade da diagnose viral baseada em testes específicos para determinar a condição sanitária da videira.

Prevalence and genetic diversity of viruses associated with rugose wood complex in Greek vineyards

Plant Disease ◽  
2021 ◽  
Author(s):  
Chrysoula Orfanidou ◽  
Kalliopi Moraki ◽  
Polina Panailidou ◽  
Leonidas Lotos ◽  
Asimina T Katsiani ◽  
...  
Keyword(s):  
Phylogenetic Trees ◽  
First Record ◽  
Grapevine Virus ◽  
Multiple Introductions ◽  
Worldwide Distribution ◽  
Grapevine Virus A ◽  
Transmission Modes ◽  
Virus Incidence ◽  
Grapevine Virus B ◽  
Rupestris Stem Pitting

Rugose wood is one of the most important disease syndromes of grapevine and it has been associated with at least three viruses: grapevine rupestris stem pitting associated virus (GRSPaV), grapevine virus A (GVA) and grapevine virus B (GVB). All three viruses show a worldwide distribution pattern, and their genetic composition has been the focus of extensive research over the past years. Despite their first record in Greece almost 20 years ago, there is a lack of knowledge on the distribution and genetic variability of their populations in Greek vineyards. In this context, we investigated the distribution of GRSPaV, GVA and GVB in rootstocks, self-rooted and grafted grapevine cultivars, originating from different geographic regions that are representing important viticultural areas of Greece. Three new RT-PCR assays were developed for the reliable detection of GRSPaV, GVA and GVB. Our results indicated that GVA is the most prevalent in Greek vineyards, followed by GRSPaV and GVB. However, virus incidence differed among self-rooted and grafted grapevine cultivars or rootstocks tested. Selected isolates from each virus were further molecularly characterized to determine their phylogenetic relationships. All three viruses exhibited high nucleotide diversity, which was depicted in the constructed phylogenetic trees. Isolates from Greece were placed in various phylogroups, reinforcing the scenario of multiple introductions of GVA, GVB and GRSPaV in Greece and highlighting the effect of different transmission modes in the evolutionary course of the three viruses.

scholarly journals Predominance and Diversity of GLRaV-3 in Native Vines of Mediterranean Croatia

Plants ◽  
2020 ◽  
Vol 10 (1) ◽  
pp. 17
Author(s):  
Katarina Hančević ◽  
Pasquale Saldarelli ◽  
Mate Čarija ◽  
Silvija Černi ◽  
Goran Zdunić ◽  
...  
Keyword(s):  
Single Strand Conformation Polymorphism ◽  
Phylogenetic Group ◽  
Enzyme Linked Immunosorbent Assay ◽  
Phylogenetic Groups ◽  
Arabis Mosaic Virus ◽  
Grapevine Virus A ◽  
Group I ◽  
Grapevine Pinot Gris Virus ◽  
Virus Genomes ◽  
Rupestris Stem Pitting

Sixteen grapevine cultivars from Mediterranean Croatia were surveyed for the presence of 10 of the most economically important grapevine viruses. The presence of Grapevine fanleaf virus (GFLV), Arabis mosaic virus (ArMV), Grapevine leafroll associated virus-1, -2, and -3 (GLRaV-1; GLRaV-2 and GLRaV-3), Grapevine virus A (GVA) and B (GVB), Grapevine fleck virus (GFkV), Grapevine rupestris stem pitting associated virus (GRSPaV), and Grapevine Pinot gris virus (GPGV) were tested by reverse transcription polymerase chain reaction (RT-PCR) and enzyme-linked immunosorbent assay (ELISA). All 71 analyzed clones were positive for the presence of one or more viruses. The most abundant one, detected in almost 95% of samples was GLRaV-3. In most of cases it was reported in mixed infections with GVA, GRSPaV, and GPGV. Virus genomes of GLRaV-3 infected vines were further characterized molecularly in order to determine their genetic diversity. Different genomic variants of heat shock 70 protein homologue (HSP70h) were identified by single-strand conformation polymorphism (SSCP) and sequenced. Sequence analysis confirmed their clustering into phylogenetic group I and/or phylogenetic group II. This study emphasizes the wide virus heterogenicity in Mediterranean vines and the predominant presence of GLRaV-3 phylogenetic groups I and II, either individually or in combination.

scholarly journals Detection and coat protein gene characterization of an isolate of Grapevine virus B from corky bark-affected grapevines in Southern Brazil

2002 ◽  
Vol 27 (3) ◽  
pp. 279-284 ◽  
Author(s):  
OSMAR NICKEL ◽  
THOR V. M. FAJARDO ◽  
FRANCISCO J. L. ARAGÃO ◽  
CÉSAR M. CHAGAS ◽  
GILMAR B. KUHN
Keyword(s):  
Coat Protein ◽  
Coat Protein Gene ◽  
Protein Gene ◽  
Amino Acid Sequences ◽  
Grapevine Virus ◽  
Leaf Extracts ◽  
Western Blots ◽  
Gene Characterization ◽  
Gene Coding ◽  
Grapevine Virus B

An isolate of Grapevine virus B (GVB), obtained by indexing Vitis labrusca and V. vinifera grapevines on the indicator LN33, was transmitted mechanically to several Nicotiana species. The virus was partially purified from N. cavicola and the coat protein estimated at 23 kDa by SDS-PAGE. In negatively stained leaf extracts of experimentally inoculated N. cavicola and N. occidentalis, flexuous particles with cross banding were observed, predominantly measuring 750-770 x 12 nm, with a modal length of 760 nm. Decoration indicated a clear, positive reaction against AS-GVB. In DAS-ELISA, GVB was detected in N. cavicola and grapevine extracts, and Western blots showed homologous and cross reaction of GVB and GVA antisera with GVB coat protein. Using specific primers for GVB, a fragment of 594 bp, comprising the coat protein gene coding for 197 amino acids, was amplified by RT-PCR with viral RNA extracted from GVB-infected N. occidentalis. The nucleotide and the deduced amino acid sequences of the coat protein gene showed high identities with Italian and Japanese isolates of GVB.

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