Fitopatologia - Artigos
URI permanente para esta coleçãohttps://locus.ufv.br/handle/123456789/11741
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Item Molecular characterization of two brazilian isolates of Lettuce mosaic virus with distinct biological properties(Fitopatologia Brasileira, 2001-06) Krause- Sakate, Renate; Mello, Raquel N.; Pavan, Marcelo A.; Zambolim, Eunize M.; Carvalho, Murilo G.; Gall, Olivier Le; Zerbini, F. Murilo; Krause-Sakate, RenateThe coat protein genes of two field isolates of Lettuce mosaic virus (LMV) from São Paulo State, previously characterized based on their virulence on lettuce (Lactuca sativa) differential cultivars as belonging to pathotypes II (isolate AF198, unable to infect cultivars possessing the genes mo11 or mo12) and IV (isolate AF199, which breaks the resistance conferred by mo11 or mo12), were cloned and sequenced. Comparisons of the nucleotide sequences from European, Middle-Eastern, North American, and the two Brazilian isolates did not distinguish strains, because homologies were always greater than 95%. However, phylogenetic analysis indicated that the Brazilian isolate AF198 clusters with isolates LMV-R and LMV-0 (pathotype II, from the United States and France, respectively). Isolate AF199 clustered with two isolates (LMV-Aud and LMV-13) from France. These isolates are also closely related to isolates from Chile, although a common origin is not proposed. Independent mutation events may be occurring in different parts of the world, leading to the emergence of distinct LMV strains capable of overcoming the resistance genes mo11 or mo12.Item Variabilidade genética de isolados de badnavírus infectando inhame (Dioscorea spp.) no Nordeste do Brasil(Tropical Plant Pathology, 2013-07) Lima, Joyce Silva; Lima, Alison T. M.; Castillo- Urquiza, Gloria P.; Silva, Sarah J. C.; Assunção, Iraildes P.; Michereff, Sami J.; Zerbini, F. Murilo; Lima, Gaus S. A.Viroses causadas por vírus do gênero Badnavirus são responsáveis por grandes prejuízos à cultura do inhame no Nordeste brasileiro. O conhecimento da variabilidade destes patógenos pode fornecer informações importantes sobre seu potencial evolutivo, permitindo a elaboração de melhores estratégias de manejo da doença. A análise de 425 amostras foliares de inhame coletadas em três estados do Nordeste brasileiro, em 2010, revelou uma alta incidência (93,3%) de badnaviroses. Para avaliar a variabilidade genética dos badnavírus infectando inhame, um fragmento de 579 nucleotídeos correspondente à região codificante da transcriptase reversa (RT)/RNaseH dos isolados amostrados foi amplificada por PCR e sequenciada. A análise filogenética das sequências de nucleotídeos revelou que os isolados dividem-se em dois grupos. Um é altamente relacionado com Dioscorea bacilliform AL virus (DBALV), enquanto o outro forma um clado altamente divergente dentro do gênero Badnavirus. Os isolados de DBALV apresentam 70-98% de identidade nucleotídica entre si e foram detectados em todas as áreas avaliadas e em D. alata e D. cayennensis, as duas espécies de inhame mais cultivadas no Nordeste. Os isolados do outro grupo compartilham 47-58% de identidade com isolados de DBALV e 78-95% entre si e foram encontrados apenas em D. alata na Paraíba.Item Caracterização de um isolado do Bean rugose mosaic virus (BRMV) de Minas Gerais e estimativa de perdas em feijoeiro em infecção simples ou em conjunto com o BCMV(Fitopatologia Brasileira, 2006-09) Maia, Felipe G. M.; Carvalho, Murilo G.; Pinto, Cleide M. F.; Zerbini, F. Murilo; Castillo-Urquiza, Gloria P.Plantas de feijão-vagem do cultivar Novirex apresentando mosaico e enrolamento de vagens, sem deformação foliar evidente, foram coletadas em 2002 em Cordisburgo, MG. Estudos preliminares identificaram o vírus como um isolado do Bean rugose mosaic virus (BRMV). Este trabalho relata a caracterização do isolado, por meio de produção e avaliação de anti-soro, determinação da gama de hospedeiros, estudo da transmissão do vírus por besouros crisomelídeos e estimativa de perdas em feijoeiro como resultado de infecção isolada ou em conjunto com o Bean common mosaic virus (BCMV). O roteiro adotado para purificação possibilitou a obtenção de vírus purificado em rendimento satisfatório para a produção de anti-soro. A titulação dos anti-soros foi realizada por ELISA indireto, obtendo-se reações positivas com a diluição máxima testada (1:70.000). Das 22 espécies vegetais utilizadas na gama de hospedeiros, foram infectadas plantas de Chenopodium quinoa e alguns cultivares de feijão e soja, conforme esperado para o BRMV. O isolado de BRMV foi transmitido pelo besouro crisomelídeo Cerotoma arcuata a uma taxa de 33,3%. A infecção simples de feijão 'Ouro Negro' e de feijão-vagem 'Novirex' levou a uma redução do peso das vagens por planta de 3,4% e 84,9%, respectivamente. Infecção mista do BRMV com o BCMV levou a uma redução do peso de vagens por planta de até 70,1% para 'Novirex' e de até 90,8% para 'Ouro Negro'.Item Efeitos na fotossíntese e área foliar de cultivares de alface inoculadas mecanicamente com patótipos do Lettuce mosaic virus e Lettuce mottle virus(Fitopatologia Brasileira, 2004-01) Jadão, Adriana S.; Pavan, Marcelo A.; Krause-Sakate, Renate; Zerbini, F. MuriloLevantamentos realizados no estado de São Paulo indicaram a ocorrência isolada e em infecções mistas do Lettuce mosaic virus (LMV) e do Lettuce mottle virus (LeMoV) em plantas de alface (Lactuca sativa). O presente trabalho teve como objetivo estudar os efeitos da infecção isolada e mista entre o LMV (patótipos II e IV) e o LeMoV, em cultivares de alface suscetível (White Boston) e tolerante (Elisa - gene mol1) ao LMV patótipo II. As plantas foram inoculadas via extrato vegetal tamponado com isolados de LMV-II, LMV-IV e LeMoV separadamente e em diferentes combinações, com intervalo de 24 h ou simultaneamente com os dois vírus. As plantas infetadas foram analisadas utilizando-se hospedeiras diferenciais para o LMV e o LeMoV, e no caso do LMV pelo teste sorológico de PTA-ELISA. Nas avaliações de peso fresco e seco, área foliar e teor de clorofila, observou-se que a cultivar White Boston foi a mais afetada por ambos os vírus. As infecções mistas e isoladas na cultivar Elisa causaram efeitos semelhantes, provavelmente devido a presença do gene mo11 de tolerância ao LMV-II. O isolado LMV-IV foi considerado o mais agressivo nestas cultivares quando comparado ao LMV-II e o LeMoV.Item Genetic diversity of begomoviruses infecting soybean, bean and associated weeds in Northwestern Argentina(Fitopatologia Brasileira, 2006-07) Rodríguez-Pardina, Patricia E.; Zerbini, F. Murilo; Ducasse, Daniel A.The subtropical Northwestern region of Argentina (provinces of Tucumán, Salta, Jujuy, Santiago del Estero and Catamarca) suffers from a high incidence of the whitefly Bemisia tabaci, and the detection of begomoviruses is also common. The Northwest is the main bean-growing region of the country, and approximately 10% of Argentina's soybean crop is grown in this area. We have used a PCR-based assay to establish the identity and genetic diversity of begomoviruses associated with bean and soybean crops in Northwestern Argentina. Universal begomovirus primers were used to direct the amplification of a fragment encompassing the 5' portion of the capsid protein gene. Amplified fragments were cloned, sequenced and subjected to phylogenetic analysis to determine the sequence identity to known begomoviruses. The data indicated the presence of four distinct begomoviruses, all related to other New World begomoviruses. The prevalent virus, which was present in 94% of bean and soybean samples and also in two weed species, is closely related to Sida mottle virus (SiMoV). A virus with high sequence identity with Bean golden mosaic virus (BGMV) was found in beans. The two remaining viruses displayed less than 89% identity with other known begomoviruses, indicating that they may constitute novel species. One of these putative novel viruses was detected in bean, soybean and tomato samples.Item Revisiting the classification of curtoviruses based on genome-wide pairwise identity(Archives of Virology, 2014-01-25) Zerbini, F. Murilo; Varsani, Arvind; Martin, Darren P.; Navas-Castillo, Jesús; Moriones, Enrique; Hernández-Zepeda, Cecilia; Idris, Ali; Brown, Judith K.Members of the genus Curtovirus (family Geminiviridae) are important pathogens of many wild and cultivated plant species. Until recently, relatively few full curtovirus genomes have been characterised. However, with the 19 full genome sequences now available in public databases, we revisit the proposed curtovirus species and strain classification criteria. Using pairwise identities coupled with phylogenetic evidence, revised species and strain demarcation guidelines have been instituted. Specifically, we have established 77 % genome-wide pairwise identity as a species demarcation threshold and 94 % genome-wide pairwise identity as a strain demarcation threshold. Hence, whereas curtovirus sequences with >77 % genome-wide pairwise identity would be classified as belonging to the same species, those sharing >94 % identity would be classified as belonging to the same strain. We provide step-by-step guidelines to facilitate the classification of newly discovered curtovirus full genome sequences and a set of defined criteria for naming new species and strains. The revision yields three curtovirus species: Beet curly top virus (BCTV), Spinach severe surly top virus (SpSCTV) and Horseradish curly top virus (HrCTV).Item Molecular mapping of the viral determinants of systemic wilting induced by a Lettuce mosaic virus (LMV) isolate in some lettuce cultivars(Virus Research, 2005-05) Zerbini, F. Murilo; Krause-Sakate, Renate; Redondo, Elise; Richard-Forget, Florence; Jadão, Adriana Salomão; German-Retana, Sylvie; Pavan, Marcelo Agenor; Candresse, Thierry; Gall, Olivier Le; Houvenaghel, Marie ChristineThe isolate AF199 of Lettuce mosaic virus (LMV, genus Potyvirus) causes local lesions followed by systemic wilting and plant death in the lettuce cultivars Ithaca and Vanguard 75. Analysis of the phenotype of virus chimeras revealed that a region within the P1 protein coding region (nucleotides 112–386 in the viral genome) and/or another one within the CI protein coding region (nucleotides 5496–5855) are sufficient together to cause the lethal wilting in Ithaca, but not in Vanguard 75. This indicates that the determinants of this particular symptom are different in these two lettuce cultivars. The wilting phenotype was not directly correlated with differences in the deduced amino acid sequence of these two regions. Furthermore, transient expression of the LMV-AF199 proteins, separately or in combination, did not induce local necrosis or any other visible reaction in the plants. Together, these results suggest that the systemic wilting reaction might be due to RNA rather than protein sequences.Item Establishment of three new genera in the family Geminiviridae: Becurtovirus, Eragrovirus and Turncurtovirus(Archives of Virology, 2014-03-22) Zerbini, F. Murilo; Varsani, Arvind; Navas-Castillo, Jesús; Moriones, Enrique; Hernández-Zepeda, Cecilia; Idris, Ali; Brown, Judith K.; Martin, Darren P.The family Geminiviridae includes plant-infecting circular single-stranded DNA viruses that have geminate particle morphology. Members of this family infect both monocotyledonous and dicotyledonous plants and have a nearly global distribution. With the advent of new molecular tools and low-cost sequencing, there has been a significant increase in the discovery of new geminiviruses in various cultivated and non-cultivated plants. In this communication, we highlight the establishment of three new genera (Becurtovirus, Eragrovirus and Turncurtovirus) to accommodate various recently discovered geminiviruses that are highly divergent and, in some cases, have unique genome architectures. The genus Becurtovirus has two viral species, Beet curly top Iran virus (28 isolates; leafhopper vector Circulifer haematoceps) and Spinach curly top Arizona virus (1 isolate; unknown vector), whereas the genera Eragrovirus and Turncurtovirus each have a single assigned species: Eragrostis curvula streak virus (6 isolates; unknown vector) and Turnip curly top virus (20 isolates; leafhopper vector Circulifer haematoceps), respectively. Based on analysis of all of the genome sequences available in public databases for each of the three new genera, we provide guidelines and protocols for species and strain classification within these three new genera.Item A genome-wide pairwise-identity-based proposal for the classification of viruses in the genus Mastrevirus (family Geminiviridae)(Archives of Virology, 2013-01-23) Zerbini, F. Murilo; Muhire, Brejnev; Martin, Darren P.; Brown, Judith K.; Navas-Castillo, Jesús; Moriones, Enrique; Rivera-Bustamante, Rafael; Malathi, V. G.; Briddon, Rob W.; Varsani, ArvindRecent advances in the ease with which the genomes of small circular single-stranded DNA viruses can be amplified, cloned, and sequenced have greatly accelerated the rate at which full genome sequences of mastreviruses (family Geminiviridae, genus Mastrevirus) are being deposited in public sequence databases. Although guidelines currently exist for species-level classification of newly determined, complete mastrevirus genome sequences, these are difficult to apply to large sequence datasets and are permissive enough that, effectively, a high degree of leeway exists for the proposal of new species and strains. The lack of a standardised and rigorous method for testing whether a new genome sequence deserves such a classification is resulting in increasing numbers of questionable mastrevirus species proposals. Importantly, the recommended sequence alignment and pairwise identity calculation protocols of the current guidelines could easily be modified to make the classification of newly determined mastrevirus genome sequences significantly more objective. Here, we propose modified versions of these protocols that should substantially minimise the degree of classification inconsistency that is permissible under the current system. To facilitate the objective application of these guidelines for mastrevirus species demarcation, we additionally present a user-friendly computer program, SDT (species demarcation tool), for calculating and graphically displaying pairwise genome identity scores. We apply SDT to the 939 full genome sequences of mastreviruses that were publically available in May 2012, and based on the distribution of pairwise identity scores yielded by our protocol, we propose mastrevirus species and strain demarcation thresholds of >78 % and >94 % identity, respectively.Item Revision of Begomovirus taxonomy based on pairwise sequence comparisons(Archives of Virology, 2015-04-18) Zerbini, F. Murilo; Brown, Judith K.; Navas-Castillo, Jesús; Moriones, Enrique; Ramos-Sobrinho, Roberto; Silva, José C. F.; Fiallo-Olivé, Elvira; Briddon, Rob W.; Hernández-Zepeda, Cecilia; Idris, Ali; Malathi, V. G.; Martin, Darren P.; Rivera-Bustamante, Rafael; Ueda, Shigenori; Varsani, ArvindViruses of the genus Begomovirus (family Geminiviridae) are emergent pathogens of crops throughout the tropical and subtropical regions of the world. By virtue of having a small DNA genome that is easily cloned, and due to the recent innovations in cloning and low-cost sequencing, there has been a dramatic increase in the number of available begomovirus genome sequences. Even so, most of the available sequences have been obtained from cultivated plants and are likely a small and phylogenetically unrepresentative sample of begomovirus diversity, a factor constraining taxonomic decisions such as the establishment of operationally useful species demarcation criteria. In addition, problems in assigning new viruses to established species have highlighted shortcomings in the previously recommended mechanism of species demarcation. Based on the analysis of 3,123 full-length begomovirus genome (or DNA-A component) sequences available in public databases as of December 2012, a set of revised guidelines for the classification and nomenclature of begomoviruses are proposed. The guidelines primarily consider a) genus-level biological characteristics and b) results obtained using a standardized classification tool, Sequence Demarcation Tool, which performs pairwise sequence alignments and identity calculations. These guidelines are consistent with the recently published recommendations for the genera Mastrevirus and Curtovirus of the family Geminiviridae. Genome-wide pairwise identities of 91 % and 94 % are proposed as the demarcation threshold for begomoviruses belonging to different species and strains, respectively. Procedures and guidelines are outlined for resolving conflicts that may arise when assigning species and strains to categories wherever the pairwise identity falls on or very near the demarcation threshold value.