RESUMO
The genetic diversity of Cymbidium mosaic virus (CymMV, family Flexiviridae) was assessed by analysing the nucleotide sequences of coat protein (CP) and partial RNA-dependent RNA polymerase (RdRp) genes. Thirty CymMV sequences from vanilla isolates, obtained in this work by direct sequencing of RT-PCR products, were compared to the sequences from ornamental orchid isolates available in GenBank. The CymMV population exhibited overall low genetic diversity (pi=0.054 and pi=0.053 for CP and RdRp genes, respectively). Phylogenetic analyses of the 85 CP and 37 RdRp sequences revealed the segregation of the isolates into two congruent monophyletic clusters; however these two subgroups did not cluster in amino sequence analysis because most of the nt mutations were synonymous. Nevertheless, the subgrouping was confirmed by highly significant Kst tests for the CP and RdRp genes. Analysis of population genetic parameters and distribution of synonymous and nonsynonymous mutations revealed that both genes were under negative selection with no recombination events. These results suggested that the CymMV isolates found in cultivated orchids worldwide have a dual origin and are expanding as if following bottlenecks.
Assuntos
Proteínas do Capsídeo/genética , Variação Genética , Potexvirus/classificação , Potexvirus/genética , RNA Polimerase Dependente de RNA/genética , Sequência de Bases , Dados de Sequência Molecular , Orchidaceae/virologia , Filogenia , Doenças das Plantas/virologia , Potexvirus/isolamento & purificação , RNA Viral/genética , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Análise de Sequência de DNA , Homologia de Sequência de Aminoácidos , Homologia de Sequência do Ácido Nucleico , Vanilla/virologiaRESUMO
Sequence was determined for the coat protein (CP) gene and 3' non-translated region (3'NTR) of two vanilla mosaic virus (VanMV) isolates from Vanilla tahitensis, respectively from the Cook Islands (VanMV-CI) and French Polynesia (VanMV-FP). Both viruses displayed distinctive features in the N-terminal region of their CPs; for VanMV-CI, a 16-amino-acid deletion including the aphid transmission-related DAG motif, and for VanMV-FP, a stretch of GTN repeats that putatively belongs to the class of natively unfolded proteins. VanMV-FP CP also has a novel DVG motif in place of the DAG motif, and an uncommon Q//V protease cleavage site. The sequences were compared to a range of Dasheen mosaic virus (DsMV) strains and to potyviruses infecting orchids. Identity was low to DsMV strains across the entire CP coding region and across the 3'NTR, but high across the CP core and the CI-6K2-NIa region. In accordance with current ICTV criteria for species demarcation within the family Potyviridae, VanMV-CI and VanMV-FP are strains of DsMV that exclusively infect vanilla.
Assuntos
Doenças das Plantas/virologia , Potyvirus/classificação , Potyvirus/genética , Vanilla/virologia , Regiões 3' não Traduzidas/genética , Motivos de Aminoácidos/genética , Sequência de Aminoácidos , Proteínas do Capsídeo/genética , Dados de Sequência Molecular , Filogenia , Polimorfismo Genético , Polinésia , Potyvirus/isolamento & purificação , Alinhamento de Sequência , Análise de Sequência de DNA , Deleção de Sequência , Homologia de SequênciaRESUMO
A survey was carried out to identify the viruses infecting vanilla in French Polynesia and to assess their incidence. Virus identification was based on enzyme-linked immunosorbent assay (ELISA) and, for potyviruses, on the sequence of part of the coat protein and inoculation assays. Between 1998 and 1999, 3,610 vanilla plants from 49 plots in the Society Islands were indexed. Cymbidium mosaic virus (CymMV) was detected in 500 vines from 10 plots in the Leeward Islands. The data suggest that this virus has spread widely since its first detection in French Polynesia in 1986, most likely through the dissemination of symptomless infected cuttings. Viruses belonging to the Potyvirus genus were found in 674 plants from 27 plots in the four islands surveyed. Three distinct potyviruses have been identified: (i) Vanilla mosaic virus, (ii) Watermelon mosaic virus, and (iii) and a virus related to Bean common mosaic virus. The symptoms induced on Vanilla tahitensis by the three potyviruses can be differentiated from each other and from those due to CymMV. A significant proportion of the plants surveyed (97/476) were symptomatic but tested negative by ELISA for CymMV and the Potyvirus group. Odontoglossum ringspot virus was not detected in any sample tested.