Molecular and biological characterization of a putative new sobemovirus infecting Physalis peruviana.

Physalis peruviana is a perennial solanaceous plant that has recently been established as a commercial crop in Brazil. This work reports the near-complete genome sequence, particle morphology, and plant host responses to a putative new sobemovirus, named "physalis rugose mosaic virus". The virus, characterized by isometric particles of ca. 30 nm in diameter, causes foliar symptoms of mosaic, malformation and blistering, accompanied by stunting. The near-complete genome sequence comprises 4175 nucleotides and contains five open reading frames that are similar to those of other sobemoviruses. In addition to P. peruviana, the new virus systemically infected Capsicum annuum, Nicotiana tabacum and Solanum lycopersicum by mechanical inoculation. Thus, this virus may cause disease in these crops in the field.

Citrus Psorosis Virus Movement Protein Contains an Aspartic Protease Required for Autocleavage and the Formation of Tubule-Like Structures at Plasmodesmata.

Plant virus cell-to-cell movement is an essential step in viral infections. This process is facilitated by specific virus-encoded movement proteins (MPs), which manipulate the cell wall channels between neighboring cells known as plasmodesmata (PD). Citrus psorosis virus (CPsV) infection in sweet orange involves the formation of tubule-like structures within PD, suggesting that CPsV belongs to "tubule-forming" viruses that encode MPs able to assemble a hollow tubule extending between cells to allow virus movement. Consistent with this hypothesis, we show that the MP of CPsV (MPCPsV) indeed forms tubule-like structures at PD upon transient expression in Nicotiana benthamiana leaves. Tubule formation by MPCPsV depends on its cleavage capacity, mediated by a specific aspartic protease motif present in its primary sequence. A single amino acid mutation in this motif abolishes MPCPsV cleavage, alters the subcellular localization of the protein, and negatively affects its activity in facilitating virus movement. The amino-terminal 34-kDa cleavage product (34KCPsV), but not the 20-kDa fragment (20KCPsV), supports virus movement. Moreover, similar to tubule-forming MPs of other viruses, MPCPsV (and also the 34KCPsV cleavage product) can homooligomerize, interact with PD-located protein 1 (PDLP1), and assemble tubule-like structures at PD by a mechanism dependent on the secretory pathway. 20KCPsV retains the protease activity and is able to cleave a cleavage-deficient MPCPsV in trans Altogether, these results demonstrate that CPsV movement depends on the autolytic cleavage of MPCPsV by an aspartic protease activity, which removes the 20KCPsV protease and thereby releases the 34KCPsV protein for PDLP1-dependent tubule formation at PD.IMPORTANCE Infection by citrus psorosis virus (CPsV) involves a self-cleaving aspartic protease activity within the viral movement protein (MP), which results in the production of two peptides, termed 34KCPsV and 20KCPsV, that carry the MP and viral protease activities, respectively. The underlying protease motif within the MP is also found in the MPs of other members of the Aspiviridae family, suggesting that protease-mediated protein processing represents a conserved mechanism of protein expression in this virus family. The results also demonstrate that CPsV and potentially other ophioviruses move by a tubule-guided mechanism. Although several viruses from different genera were shown to use this mechanism for cell-to-cell movement, our results also demonstrate that this mechanism is controlled by posttranslational protein cleavage. Moreover, given that tubule formation and virus movement could be inhibited by a mutation in the protease motif, targeting the protease activity for inactivation could represent an important approach for ophiovirus control.

Serial analysis of gene expression (SAGE) of host-plant resistance to the cassava mosaic disease (CMD).

Cassava mosaic disease (CMD) is a viral disease of the important tropical staple crop cassava (Manihot esculenta) and preferred management involves use of host-plant resistance. The best available resistance is controlled by a single dominant gene. Serial analysis of gene expression (SAGE) was used to analyze the gene expression pattern in a bulk of 40 each of CMD resistant and susceptible genotypes drawn from a gene mapping progeny. Messenger RNA used for the SAGE analysis came from plants that were exposed to heavy disease pressure over a period of 2 years in the field. A total of 12,786 tags were studied, divided into 5733 and 7053 tags from the resistant and susceptible genotypes, respectively. Tag annotation was by PCR amplification using the tag sequence as sense primer and 4000 cassava ESTs generated from the bulk of CMD resistant genotypes. Annotation of more than 30 differentially expressed tags revealed several genes expressed during systemic acquired resistance (SAR) in plants and other genes involved in cell-to-cell and cytoplasm-to-nucleus virus trafficking. Differential expression of the most abundantly expressed tag, corresponding to a beta-tubulin gene, was confirmed by Northern Analysis. RFLP analysis of the tags in the parents and bulks of the CMD mapping progeny revealed only one tag, a WRKY transcription factor, associated with the region bearing the dominant CMD gene.

Factors favoring houseplant container infestation with Aedes aegypti larve in Marilia, Sao Paulo, Brazil / Factores que favorecen la infestación de recipientes con plantas domésticas por larvas de Aedes aegypti en Marília, São Paulo, Brasil

Desde 1985, después de la reinfestación de mosquitos Aedes aegypti (L.) en el Estado de São Paulo, se encontró que ­a pesar de la diseminación de mensajes educativos destinados a su control­ las macetas y floreros son un hábitat importante de las larvas. Los objetivos de este estudio fueron identificar las macetas y floreros como hábitats de larvas de mosquitos en relación con su número y grado de infestación, los tipos de plantas involucradas y la localización específica de las larvas; investigar los nombres locales de las plantas domésticas y examinar los factores que afectan al cumplimiento de las medidas de control. Los resultados mostraron un promedio de más de cuatro posibles hábitats de larvas relacionados con plantas por cada sitio, de los cuales solo 0,4% tenían el vector. Los recipientes para plantas representaron 31% de todos los receptáculos con larvas de A. aegypti. A pesar de que de una muestra de 126 personas entrevistadas enumeraron 105 nombres de plantas domésticas, 49% de las plantas asociadas con larvas fueron solo de dos tipos: helechos y la planta ornamental Dieffenbachia avoena. La evidente falta de voluntad de la población para aceptar las medidas de control contra A. aegypti en lo que respecta a plantas domésticas parece deberse a la poca frecuencia relativa de A. aegypti en los recipientes domésticos, la falta de confianza en los programas de control como fuente idónea de información sobre plantas y la opinión de que las medidas de control recomendadas son dañinas para las plantas. Actualmente se está planeando una intervención para el control del dengue en la que se usará material educativo que trata específicamente sobre las plantas cuyos recipientes albergan con mayor frecuencia las larvas de A. aegypti. También se hará uso de fuentes de información botánica con mayor credibilidad en materia de plantas. Además, se formulará un plan diferente con un enfoque más atractivo desde el punto de vista de la salud de las plantas para incrementar su posibilidad de aceptación.

Desde 1985, después de la reinfestación de mosquitos Aedes aegypti (L.) en el Estado de São Paulo, se encontró que ­a pesar de la diseminación de mensajes educativos destinados a su control­ las macetas y floreros son un hábitat importante de las larvas. Los objetivos de este estudio fueron identificar las macetas y floreros como hábitats de larvas de mosquitos en relación con su número y grado de infestación, los tipos de plantas involucradas y la localización específica de las larvas; investigar los nombres locales de las plantas domésticas y examinar los factores que afectan al cumplimiento de las medidas de control. Los resultados mostraron un promedio de más de cuatro posibles hábitats de larvas relacionados con plantas por cada sitio, de los cuales solo 0,4% tenían el vector. Los recipientes para plantas representaron 31% de todos los receptáculos con larvas de A. aegypti. A pesar de que de una muestra de 126 personas entrevistadas enumeraron 105 nombres de plantas domésticas, 49% de las plantas asociadas con larvas fueron solo de dos tipos: helechos y la planta ornamental Dieffenbachia avoena. La evidente falta de voluntad de la población para aceptar las medidas de control contra A. aegypti en lo que respecta a plantas domésticas parece deberse a la poca frecuencia relativa de A. aegypti en los recipientes domésticos, la falta de confianza en los programas de control como fuente idónea de información sobre plantas y la opinión de que las medidas de control recomendadas son dañinas para las plantas. Actualmente se está planeando una intervención para el control del dengue en la que se usará material educativo que trata específicamente sobre las plantas cuyos recipientes albergan con mayor frecuencia las larvas de A. aegypti. También se hará uso de fuentes de información botánica con mayor credibilidad en materia de plantas. Además, se formulará un plan diferente con un enfoque más atractivo desde el punto de vista de la salud de las plantas para incrementar su posibilidad de aceptación.

Antiviral activity of S-adenosylhomocysteine hydrolase inhibitors against plant viruses.

Three SAH hydrolase inhibitors, (RS)-3-adenin-9-yl-2-hydroxypropanoic acid (isobutyl ester) [(RS)-AHPA]; (RS)-9-(2,3-dihydroxypropyl)adenine [(RS)-DHPA] and the carbocyclic analog of 3-deazaadenosine (C-c3Ado) were evaluated for their inhibitory activity against tobacco mosaic virus (TMV) and potato virus X (PVX). Using the local lesion assay and ELISA, we demonstrated that all three compounds inhibit the replication of TMV and PVX. Whereas the three compounds proved about equally active against PVX, (RS)-AHPA was the most effective against TMV. (RS)-AHPA and C-c3Ado induced chlorosis in Nicotiana tabacum leaf discs. They also caused a substantial reduction in the growth of the main root of Phaseolus vulgaris. (RS)-DHPA was less phytotoxic than its two congeners.