RESUMO
Total RNAs from a symptomless tomato plant were subjected to next-generation sequencing (NGS) analysis, revealing the presence of a single viral agent - potato virus Y (PVY). The analysis of determined full-length genome sequence assigned the PVY SL16 isolate to the recombinant PVY-N-Wi strain group. A series of primers targeting the four main recombinant junction (RJ) sites were used for characterization of additional 5 tomato PVY isolates recovered in Western Slovakia. Based on the partial sequences, the isolates could be classified as belonging to PVY-N-Wi and PVY-NTNa strain groups. Interestingly, both these distinct recombinant PVY types were identified in mixed infection in one tomato sample (SL31). Our results further reinforce the data on the complexity of PVY infection and confirm the recombination as a significant evolutionary process shaping the PVY diversity.
Assuntos
Doenças das Plantas/virologia , Potyvirus/genética , Potyvirus/isolamento & purificação , Recombinação Genética , Solanum lycopersicum/virologia , Proteínas do Capsídeo/genética , Primers do DNA/genética , Genoma Viral , Filogenia , Potyvirus/classificação , RNA Viral/genética , Análise de Sequência de DNA , EslováquiaRESUMO
Although grapevine leafroll-associated virus 1 (GLRaV-1) is one of the most important agents of the grapevine leafroll disease, the data about its molecular variability are scarce. In order to assess the GLRaV-1 diversity in Slovakia, the sequence of a genome region encoding the central part of the capsid protein (CP) gene was determined from 18 GLRaV-1 isolates. Despite the fact that the analysis targeted a relatively short portion of the genome, comparison of obtained sequences has revealed the nucleotide identities between Slovak isolates ranging from 83.0-100%. Phylogenetic analysis indicated the presence of two distinct molecular groups of GLRaV-1 in Slovakia.
Assuntos
Closteroviridae/genética , Closteroviridae/isolamento & purificação , Genoma Viral , Doenças das Plantas/virologia , Vitis/virologia , Sequência de Aminoácidos , Proteínas do Capsídeo/química , Proteínas do Capsídeo/genética , Closteroviridae/química , Closteroviridae/classificação , Dados de Sequência Molecular , Filogenia , Alinhamento de Sequência , Eslováquia , Vitis/classificaçãoRESUMO
UNLABELLED: Plum pox virus (PPV) is the causal agent of Sharka, considered to be the most detrimental viral disease of Prunus spp. worldwide. So far, several PPV strains have been recognized, three of them (PPV-D, PPV-M, and PPV-Rec) having shown serious economic impact in the European area. Infectious cDNA clones of plant RNA viruses are excellent tools for functional studies of viral genomes. Preparation and use of PPV-D and PPV-M infectious clones have been previously reported. Here we describe the construction of an infectious cDNA clone of the strain PPV-Rec (isolate BOR-3) by the strategy involving the subsequent exchanges of homologous BOR-3 genome parts in the backbone of the previously prepared PPV-D infectious construct. The infectivity of each intermediate chimeric cDNA as well as that of the final construct (pIC-PPV-Rec) was confirmed by biolistic transfection of Nicotiana benthamiana plants. Complete sequence of the cloned viral BOR-3 cDNA revealed 0.14% of difference at the nucleotide level compared to original BOR-3 sequence, resulting in four amino acid changes. This slight inequality was related to the population heterogeneity of the initial BOR-3 isolate; no difference in the amino acid sequence resulted from the cloning steps performed. KEYWORDS: inter-strain chimera; biolistics; genome sequence.
Assuntos
DNA Complementar , Vírus Eruptivo da Ameixa , Clonagem Molecular , Genoma Viral , Doenças das Plantas/virologia , Vírus Eruptivo da Ameixa/genética , Prunus/virologia , Prunus domesticaRESUMO
Simple, fast, low-cost, and efficient procedure for DNA delivery to the cell nuclei of whole plants was developed. The procedure was optimized for the Plum pox virus (PPV) and its host Nicotiana benthamiana. It is based on the leaf bombardment with tungsten microparticles with bound DNA using common air gun. The procedure did not require special equipment and source of driving gas. The transfection efficiency obtained by the newly developed procedure was close to 100%, but this level dropped with the age of bombarded plants.
Assuntos
Biolística/métodos , DNA Complementar/genética , Nicotiana/genética , Vírus Eruptivo da Ameixa/genética , Transfecção/métodos , Biolística/instrumentação , DNA Complementar/metabolismo , Regulação da Expressão Gênica de Plantas , Doenças das Plantas/virologia , Plantas Geneticamente Modificadas/genética , Plantas Geneticamente Modificadas/virologia , Vírus Eruptivo da Ameixa/fisiologia , Nicotiana/virologia , Transfecção/instrumentaçãoRESUMO
Cucurbit aphid-borne yellows virus (CABYV) causes yellowing symptoms in cucurbit crops worldwide. In this work, the sequences for the coat protein (CP) gene for CABYV isolates from Iran and Slovakia are reported for the first time. Three Iranian isolates shared 96.2-99.3% and the Slovak isolate 98.9% of nucleotide identity in comparison with the reference French CABYV-N isolate. Phylogenetic analysis showed that the sequence of CP gene of examined CABYV isolates differed only slightly. Relatively close relationship and sequence similarity of geographically distant CABYV isolates could reflect the epidemic outbreak and rapid expansion of the virus throughout the world due to the host and vector abundance.