TRANSGENIC GRAPEVINES EXPRESSING COAT PROTEIN GENE SEQUENCES OF GRAPEVINE FANLEAF VIRUS, ARABIS MOSAIC VIRUS, GRAPEVINE VIRUS A AND GRAPEVINE VIRUS B

<p style="text-align: justify;">Sanitary status of visually selected and nonselected indigenous grapevine cultivar 'Refosk' (Vitis vinifera L. cv. 'Refosk') was evaluated. Nine viruses were tested by ELISA: Nepoviruses Grapevine fanleaf virus (GFLV) and Arabis mosaic virus (ArMV), Closteroviruses Grapevine leafroll associated viruses 1 (GLRaV-1), 2 (GLRaV-2), 3 (GLRaV-3), and 6 (GLRaV-6), Grapevine fleck virus (GFkV) and Vitiviruses Grapevine virus A (GVA) and Grapevine virus B (GVB). All viruses except ArMV and GVB were detected. The vines of cv. ‘Refosk’ passing the visual selection were 50 % free of tested viruses in comparison to only 24 % virus-free vines of non-visually selected material. Visually selected plants revealed the highest incidence of virus GLRaV- 1 (37 %) and in addition to that a 15 % incidence of rugose wood (RW) disease symptoms.</p>

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

Ciência Rural ◽

10.1590/s0103-84782010001100001 ◽

2010 ◽

Vol 40 (11) ◽

pp. 2249-2255 ◽

Cited By ~ 9

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.

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

Fitopatologia Brasileira ◽

10.1590/s0100-41582002000300007 ◽

2002 ◽

Vol 27 (3) ◽

pp. 279-284 ◽

Cited By ~ 9

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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