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• W1926309145 abstract "Brugmansia suaveolens mottle virus (BsMoV) was isolated from a Brugmansia suaveolens plant (known as angel trumpet) in the collection of medicinal plants of the Instituto Agronomico, Campinas-SP, Brazil, being the isolate named Bs-Campinas. A previous study showed that the isolate BsCampinas was transmitted by aphids and induced visible symptom on some Solanaceous plants and two Chenopodium species under the experimental conditions, showing typical potyvirus particles and inclusions in symptomatic leaves, when observed by transmission electron microscopy. Only part of the virus genome was known, a region comprising the 3' terminal region of the genome, which distinguished it sufficiently from other viruses to be classified as a new potvvirus species. The virus showed characteristics that were distinct from other known potyviruses, such as the high symptom severity in many plants and the ability to infect tomato plants, a crop of high economic and social importance in Brazil. These findings showed that it can be an excellent target for the study of genes related to pathogenicity and for the future development of viral vectors. Therefore, the objective of this study was the molecular characterization of the isolate Bs-Campinas through the determination of its complete genomic sequence and its phylogenetic analysis, when compared with different potyviruses previously reported in the world. For this purpose, some techniques were attempted to enable the rescue of the viral genome and subsequent determination of its sequence. Total RNA was extracted and cDNA synthesis was done by 3 'RACE using universal primers for potyviruses, resulting in the amplification of a fragment of ca. 8,3 kb. However, the cloning of this fragment resulted in clones that presented an internal deletion in this fragment. Therefore, a second fragment using specific primers designed based on the sequence ends of the deleted clones, was amplified by PCR and cloned in order to rescue inner region of the genome by generating a fragment of approximately 4.7 kb. Specific primers were also designed based on the 5‟ end region of the 8,3 kb clone to enable cloning of the 5‟ terminal region of the genome by 5' RACE, a fragment of 1.8 kb. PCR amplifications resulted in three cDNA fragments, comprising the complete genome of BsMoV. The RNA genome consisted of 9870 nt without a poly-A tail, encoding a putative typical potyviral polyprotein of 3090 aa. Analysis of the amino acid sequence allowed the deduction of the cleavage sites, as well as the identification of conserved sequences and motifs within the sequence of each protein, in which the BsCampinas isolate displayed typical potyvirus genome strategy. Phylogenetic analysis, as well as comparisons with other species of the Potyvirus genus obtained on online databases revealed that this BsMoV sequence shared 63.7% nucleotide sequence with Pepper mottle virus (PepMoV), which was the best matched potyvirus sequence and, thus the most closely related species. Since this identity value is below the threshold of 76% nt identity presently used to discriminate Potyvirus species, it was confirmed that Brugmansia suaveolens mottle virus represents a new Potyvirus species." @default.
• W1926309145 created "2016-06-24" @default.
• W1926309145 creator A5049361397 @default.
• W1926309145 date "2010-10-28" @default.
• W1926309145 modified "2023-09-26" @default.
• W1926309145 title "Determinação da sequência do genoma completo do potyvirus Brugmansia suaveolens mottle virus" @default.
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• W1926309145 hasPublicationYear "2010" @default.
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