Detección del virus de la leprosis de los cítricos tipo citoplasmático en los Llanos Orientales de Colombia.

Durante los años 2004 y 2005 se observaron síntomas de la leprosis de los cítricos en los departamentos de Meta y Casanare. Para confirmar la causa de esta sintomatología, se colectaron hojas de naranja 'Valencia' con lesiones típicas en municipios de Casanare (Yopal y Aguazul) y Meta (Guamal, Villavicencio y Cumaral). Las muestras fueron procesadas identificar el virus por medio de microscopía electrónica de transmisión (MET) y transcripción reversa - reacción en cadena de la polimerasa (RT-PCR). Diez de las 21 muestras analizadas por MET resultaron positivas para el virus de la leprosis (Citrus Leprosis Virus, CiLV-C). En algunas muestras se observaron partículas baciliformes características del CiLV-C en el retículo endoplasmático y formas irregularles de viroplasmas en el citoplasma. Catorce de las 32 muestras procesadas por RT-PCR resultaron positivas al CiLV-C. Los análisis RT-PCR amplificaron fragmentos de cADN del tamaño esperado para las muestras colectadas en el Meta. Una de las ampliaciones fue secuenciada (GenBank accessión No. DQ272491) y la secuencia obtenida guarda identidad del 98 por ciento con la secuencia brasilera aislada de nucleótidos para CiLV-C (GenBank accessión No. AY289190.1). La secuencia obtenida y los primers fueron entregados al Instituto Colombiano Agropecuario -ICA- para el diagnóstico del virus CiLV-C en Colombia. Entre los ácaros recolectados en las plantas afectadas se identificó a Brevipalpus phoenicis (Geijskes) reconocido como principal vector de CiLV-

Levantamento preliminar de viroses de plantas cultivadas nos arredores de Manaus, Am

Um levantamento preliminar de viroses de plantas cultivadas nos arredores de Manaus foi feito em março de 1977. Diversas propriedades particulares e campos experimentais foram inspecionados, anotando-se as plantas exibindo sintomatologia suspeita. A verificação da sua possível etiologia viral foi feita através de testes de transmissão, serologia e exames electrono microscópicos. Foram constatadas diversas viroses em tomateiro, pimentão, quiabeiro, feijoeiro, cucurbitáceas, soja, feijão-de-asa, milho. etc. Como regra geral, ao contrário do que sucede no Sul do País, as viroses transmitidas por afídeos foram de ocorrência rara, presumivelmente porque as condições climáticas da região seriam desfavoráveis à biologia desses insetos

Identificação e purificação de um vírus-de-granulose em lagartas-do-cartucho-do-milho

A virus was found infecting larvae of fall armyworm, Spodoptera frugiperda (Lepidoptera: Noctuidae) (Smith, 1797), in Sete Lagoas, MG, Brazil. Virus extracts from infected larvae were able to cause up to 100% mortality in three-day old larvae feeding in artificial diet. The identification of the virus, made by electron microscopy, showed that it was a granulosis virus, belonging to the genus Baculovirus and is composed of enveloped nucleocapsids which are occluded in proteinaceous inclusion bodies. Purifications by differential and sucrose gradient centrifugations yielded 22 mg of inclusion bodies per infected larva. Due to its pathogenicity, large number of inclusion bodies per infected larvae and simplicity of purification procedures, the identified virus shows a great potential to be used as a bioinsecticide in the biological control of the fall armyworm. Foi constatada a presença de um vírus-de-granulose (VG) infectando lagartas-do-cartucho-do-milho, Spodoptera frugiperda (Lepidoptera: Noctuidae) (Smith, 1797), VGSf, na região de Sete Lagoas, MG. Extratos de lagartas infectadas com vírus mostraram ser patogênicos, chegando a causar até 100% de mortalidade em lagartas de três dias de idade, criadas artificialmente em laboratório. A identificação do vírus foi feita através de microscopia eletrônica, e os resultados mostraram tratar-se de um vírus-de-granulose, o qual pertence ao género Baculovirus e caracteriza-se por apresentar suas partículas oclusas individualmente em uma cápsula de proteína, formando estruturas características chamadas "corpos de inclusão" (CIs). A purificação do vírus, feita através de centrifugações diferenciais e em gradientes de sacarose, mostrou ser possível obter cerca de 22 mg de CIs do vírus por lagarta infectada. Dada a sua patogenicidade, grande quantidade de CIs por lagarta infectada e facilidade de purificação, o vírus em estudo apresenta um grande potencial para ser utilizado como bioinseticida no controle da lagarta-do-cartucho.

Citrus leprosis virus C infection results in hypersensitive-like response, suppression of the JA/ET plant defense pathway and promotion of the colonization of its mite vector

Leprosis is a serious disease of citrus caused by Citrus leprosis virus C (CiLV-C, genus Cilevirus) whose transmission is mediated by false spider mites of the genus Brevipalpus. CiLV-C infection does not systemically spread in any of its known host plants, thus remaining restricted to local lesions around the feeding sites of viruliferous mites. To get insight into this unusual pathosystem, we evaluated the expression profiles of genes involved in defense mechanisms of Arabidopsis thaliana and Citrus sinensis upon infestation with non-viruliferous and viruliferous mites by using reverse-transcription qPCR. These results were analyzed together with the production of reactive oxygen species (ROS) and the appearance of dead cells as assessed by histochemical assays. After interaction with non-viruliferous mites, plants locally accumulated ROS and triggered the salicylic acid (SA) and jasmonate/ethylene (JA/ET) pathways. ERE branch of the JA/ET pathways was highly activated. In contrast, JA pathway genes were markedly suppressed upon the CiLV-C infection mediated by viruliferous mites. Viral infection also intensified the ROS burst and cell death, and enhanced the expression of genes involved in the RNA silencing mechanism and SA pathway. After 13 days of infestation of two sets of Arabidopsis plants with non-viruliferous and viruliferous mites, the number of mites in the CiLV-C infected Arabidopsis plants was significantly higher than in those infested with the non-viruliferous ones. Oviposition of the viruliferous mites occurred preferentially in the CiLV-C infected leaves. Based on these results, we postulated the first model of plant/Brevipalpus mite/cilevirus interaction in which cells surrounding the feeding sites of viruliferous mites typify the outcome of a hypersensitive-like response, whereas viral infection induces changes in the behavior of its vector7CONSELHO NACIONAL DE DESENVOLVIMENTO CIENTÍFICO E TECNOLÓGICO - CNPQFUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULO - FAPESP573848/08-4; 375843/2012-42012/18771- 0; 2014/00366-8; 2008/57909-2; 2014/08458-

Alta incidência de Tomato chlorotic spot virus em jiló no estado de São Paulo

Em processament

Ocorrência do pepper yellow mosaic virus e cucumber mosaic virus em Capsicum chinense no estado do Amazonas, Brasil

The habanero chilli pepper, Capsicum chinense is an important crop in the Amazon Basin, mainly grown by small-scale producers. Capsicum chinense plants in an experimental field in the northern Brazilian state of Amazonas were found exhibiting characteristic symptoms of viral infection. Leaf sap from symptomatic plants examined under a transmission electron microscope revealed the presence of elongated flexuous particles and isometric particles. Using molecular assays, the viruses were identified as pepper yellow mosaic virus (PepYMV) and cucumber mosaic virus (CMV). Aphids, identified as Aphis gossypii, were found colonizing the C. chinense plants in the field and may be the vector for both PepYMV and CMV. We report the first occurrence of these viruses infecting C. chinense in the state of Amazonas.A pimenta-de-cheiro, Capsicum chinense é uma cultura importante na Bacia Amazônica, cultivada principalmente por pequenos produtores. Plantas de C. chinense em um campo experimental localizado no norte do estado brasileiro do Amazonas, foram encontradas apresentando sintomas característicos de infecção viral. Extratos de amostras de folhas sintomáticas examinados ao microscópio eletrônico de transmissão revelaram a presença de partículas alongadas e flexuosas e de partículas isométricas. Análises moleculares permitiram identificar a presença do pepper yellow mosaic virus (PepYMV) e do cucumber mosaic virus (CMV). Pulgões, identificados como Aphis gossypii foram encontrados colonizando pimenteiras-de-cheiro neste campo experimental e podem representar o provável vetor de PepYMV e CMV. Este trabalho relata a primeira ocorrência desses vírus infectando C. chinense no estado do Amazonas

A betabaculovirus encoding a gp64 homolog

Abstract\ud \ud Background\ud A betabaculovirus (DisaGV) was isolated from Diatraea saccharalis (Lepidoptera: Crambidae), one of the most important insect pests of the sugarcane and other monocot cultures in Brazil.\ud \ud \ud Results\ud The complete genome sequence of DisaGV was determined using the 454-pyrosequencing method. The genome was 98,392 bp long, which makes it the smallest lepidopteran-infecting baculovirus sequenced to date. It had a G + C content of 29.7 % encoding 125 putative open reading frames (ORF). All the 37 baculovirus core genes and a set of 19 betabaculovirus-specific genes were found. A group of 13 putative genes was not found in any other baculovirus genome sequenced so far. A phylogenetic analysis indicated that DisaGV is a member of Betabaculovirus genus and that it is a sister group to a cluster formed by ChocGV, ErelGV, PiraGV isolates, ClanGV, CaLGV, CpGV, CrleGV, AdorGV, PhopGV and EpapGV. Surprisingly, we found in the DisaGV genome a G protein-coupled receptor related to lepidopteran and other insect virus genes and a gp64 homolog, which is likely a product of horizontal gene transfer from Group 1 alphabaculoviruses.\ud \ud \ud Conclusion\ud DisaGV represents a distinct lineage of the genus Betabaculovirus. It is closely related to the CpGV-related group and presents the smallest genome in size so far. Remarkably, we found a homolog of gp64, which was reported solely in group 1 alphabaculovirus genomes so far.BTP received a scholarship from CAPES through the Graduate Program in\ud Biotechnology from the University of São Paulo and Butantan Institute. This project was partially funded by CNPq grant 481741/2008-9. We thank Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) and Fundação de Apoio à Pesquisa do Distrito Federal (FAPDF)