Análisis filogenético de aislamientos de Groundnut ringspot virus desde maní e identificación de posibles trips vectores asociados al cultivo de maní en la Argentina

Groundnut ringspot virus (GRSV), genus Tospovirus, is a thrips-transmitted virus infecting peanuts (Arachis hypogaea L.) in Córdoba province, Argentina. Fourteen viral isolates were recovered from Tospovirus-like symptomatic plants from different peanut fields. Viral isolates as GRSV were identified by serological and molecular tests. Nucleotide and derived amino acid sequence analyses of the nucleocapsid (N) gene indicated a high degree of identity between the GRSV peanut isolates, indicating that there is no molecular variability in the N gene of the GRSV that infects peanuts in the cropping area of Córdoba. In this study, we determined the presence of thrips species in the crop, which can potentially transmit the virus. Thrips were observed in all the evaluated peanut fields. Frankliniella schultzei was the most frequently identified species followed by Caliothrips phaseoli and Frankliniella occidentalis. This work reports the presence of F. schultzei and F. occidentalis in peanuts in Argentina for the first time. These results along with the high degree of similarity between the GRSV peanut isolates suggest that the virus could be transmitted by F. schultzei, which has been cited as its most efficient vector.Groundnut ringspot virus (GRSV, género Tospovirus) es un virus que infecta naturalmente el cultivo de maní (Arachis hypogaea L.) en la región productora de Córdoba, Argentina. En distintas localidades de la provincia, se colectaron 14 aislamientos virales provenientes de maníes que manifestaban síntomas característicos de Tospovirus. Todos los aislamientos virales fueron identificados como GRSV mediante pruebas serológicas y moleculares. El análisis de las secuencias nucleotídicas y de amino ácidos deducidas del gen de la nucleoproteína (N) reveló un alto grado de identidad entre los 14 aislamientos, indicando que no existe variabilidad molecular en el gen N del GRSV que infecta maní en la provincia de Córdoba. En este estudio se determinó la presencia de trips en el cultivo que pueden potencialmente transmitir la enfermedad. Estos insectos fueron observados colonizando maní en todos los lotes evaluados. La especie identificada con mayor frecuencia fue Frankliniella schultzei, seguida de Caliothrips phaseoli y Frankliniella occidentalis. Este es el primer reporte de F. schultzei y F. occidentalis afectando maní en Argentina. Estos resultados, junto con el elevado grado de similitud encontrado entre los distintos aislamientos de GRSV, sugieren que el virus puede ser transmitido por F. schultzei, citado como el vector más eficiente del GRSV

Recognition and blocking of innate immunity cells by Candida albicans chitin

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Sequence diversity in coat protein of SCMV infecting maize and sorghum in Brazil.

The 'maize common mosaic', caused by potyvirus, is among the major virus diseases of this crop in Brazil. Although there were evidences indicating Sugarcane mosaic virus (SCMV) as the most common potyvirus species in maize (Zea mays L.) in Brazil, information about those species that infect sorghum plants [Sorghum bicolor(L.) Moench] are few. Leaves showing characteristic mosaic symptoms were collected from maize and sorghum and used in serological and sequencing analysis of the coat protein (CP) gene for potyvirus species identification. Amino acid (aa) analysis of the CP N-terminal sequence of our samples showed a different repeated sequence, a higher content of the dipeptide GT, and a 15 aa longer than the majority of the SCMV sequences used for comparisons. The Brazilian maize and sorghum potyviruses formed a monophyletic group, suggesting that they can be classified within a new SCMV strain. Studies using potyvirus CP gene sequencing from Brazilian sorghum potyvirus have been reported for the first time

Análise do N-terminal da proteína capsidial de SCMV infectando milho e sorgo no Brasil.

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Sequenciamento do gene da capa protéica da SCMV causando sintomas de mosaico em milho, sorgo e cana de açúcar no Brasil.

xSuplemento. Edição dos Resumos do XXXIX Congresso Brasileiro de Fitopatologia, Salvador, BA, ago. 2006

The Rewiring of Ubiquitination Targets in a Pathogenic Yeast Promotes Metabolic Flexibility, Host Colonization and Virulence

Funding: This work was funded by the European Research Council [http://erc.europa.eu/], AJPB (STRIFE Advanced Grant; C-2009-AdG-249793). The work was also supported by: the Wellcome Trust [www.wellcome.ac.uk], AJPB (080088, 097377); the UK Biotechnology and Biological Research Council [www.bbsrc.ac.uk], AJPB (BB/F00513X/1, BB/K017365/1); the CNPq-Brazil [http://cnpq.br], GMA (Science without Borders fellowship 202976/2014-9); and the National Centre for the Replacement, Refinement and Reduction of Animals in Research [www.nc3rs.org.uk], DMM (NC/K000306/1). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. Acknowledgments We thank Dr. Elizabeth Johnson (Mycology Reference Laboratory, Bristol) for providing strains, and the Aberdeen Proteomics facility for the biotyping of S. cerevisiae clinical isolates, and to Euroscarf for providing S. cerevisiae strains and plasmids. We are grateful to our Microscopy Facility in the Institute of Medical Sciences for their expert help with the electron microscopy, and to our friends in the Aberdeen Fungal Group for insightful discussions.Peer reviewedPublisher PD

Stimulation of Chitin Synthesis Rescues Candida albicans from Echinocandins

Echinocandins are a new generation of novel antifungal agent that inhibit cell wall β(1,3)-glucan synthesis and are normally cidal for the human pathogen Candida albicans. Treatment of C. albicans with low levels of echinocandins stimulated chitin synthase (CHS) gene expression, increased Chs activity, elevated chitin content and reduced efficacy of these drugs. Elevation of chitin synthesis was mediated via the PKC, HOG, and Ca2+-calcineurin signalling pathways. Stimulation of Chs2p and Chs8p by activators of these pathways enabled cells to survive otherwise lethal concentrations of echinocandins, even in the absence of Chs3p and the normally essential Chs1p, which synthesize the chitinous septal ring and primary septum of the fungus. Under such conditions, a novel proximally offset septum was synthesized that restored the capacity for cell division, sustained the viability of the cell, and abrogated morphological and growth defects associated with echinocandin treatment and the chs mutations. These findings anticipate potential resistance mechanisms to echinocandins. However, echinocandins and chitin synthase inhibitors synergized strongly, highlighting the potential for combination therapies with greatly enhanced cidal activity

Eosinophils Are Recruited in Response to Chitin Exposure and Enhance Th2-Mediated Immune Pathology in Aspergillus fumigatus Infection

In patients infected with the fungus Aspergillus fumigatus, Th1 responses are considered protective, while Th2 responses are associated with increased morbidity and mortality. How host-pathogen interactions influence the development of these protective or detrimental immune responses is not clear. We compared lung immune responses to conidia from two fungal isolates that expressed different levels of the fungal cell wall component chitin. We observed that repeated aspirations of the high-chitin-expressing isolate Af5517 induced increased airway eosinophilia in the lungs of recipient mice compared to the level of eosinophilia induced by isolate Af293. CD4+ T cells in the bronchoalveolar lavage fluid (BALF) of Af5517-aspirated mice displayed decreased gamma interferon secretion and increased interleukin-4 transcription. In addition, repeated aspirations of Af5517 induced lung transcription of the Th2-associated chemokines CCL11 (eotaxin-1) and CCL22 (macrophage-derived chemokine). Eosinophil recruitment in response to conidial aspiration was correlated with the level of chitin exposure during germination and was decreased by constitutive lung chitinase expression. Moreover, eosinophil-deficient mice subjected to multiple aspirations of Af5517 prior to neutrophil depletion and infection exhibited decreased morbidity and fungal burden compared to the levels of morbidity and fungal burden found in wild-type mice. These results suggest that exposure of chitin in germinating conidia promotes eosinophil recruitment and ultimately induces Th2-skewed immune responses after repeated aspiration. Furthermore, our results suggest that eosinophils should be examined as a potential therapeutic target in patients that mount poorly protective Th2 responses to A. fumigatus infection

Participation of Candida albicans transcription factor Rlm1 in cell wall biogenesis and virulence

Candida albicans cell wall is important for growth and interaction with the environment. RLM1 is one of the putative transcription factors involved in the cell wall integrity pathway, which plays an important role in the maintenance of the cell wall integrity. In this work we investigated the involvement of RLM1 in the cell wall biogenesis and in virulence. Newly constructed C. albicans Δ/Δrlm1 mutants showed typical cell wall weakening phenotypes, such as hypersensitivity to Congo Red, Calcofluor White, and caspofungin (phenotype reverted in the presence of sorbitol), confirming the involvement of RLM1 in the cell wall integrity. Additionally, the cell wall of C. albicans Δ/Δrlm1 showed a significant increase in chitin (213%) and reduction in mannans (60%), in comparison with the wild-type, results that are consistent with cell wall remodelling. Microarray analysis in the absence of any stress showed that deletion of RLM1 in C. albicans significantly down-regulated genes involved in carbohydrate catabolism such as DAK2, GLK4, NHT1 and TPS1, up-regulated genes involved in the utilization of alternative carbon sources, like AGP2, SOU1, SAP6, CIT1 or GAL4, and genes involved in cell adhesion like ECE1, ALS1, ALS3, HWP1 or RBT1. In agreement with the microarray results adhesion assays showed an increased amount of adhering cells and total biomass in the mutant strain, in comparison with the wild-type. C. albicans mutant Δ/Δrlm1 strain was also found to be less virulent than the wild-type and complemented strains in the murine model of disseminated candidiasis. Overall, we showed that in the absence of RLM1 the modifications in the cell wall composition alter yeast interaction with the environment, with consequences in adhesion ability and virulence. The gene expression findings suggest that this gene participates in the cell wall biogenesis, with the mutant rearranging its metabolic pathways to allow the use of alternative carbon sources.This work was supported by CBMA (Centre of Molecular and Environmental Biology) through the FCT (Fundacao para a Ciencia e Tecnologia) project PEst-C/BIA/UI4050/2011. Yolanda Delgado-Silva was supported by an ALbAN scholarship (No E07D400922PE), and Alexandra Correia by SFRH/BD/31354/2006 fellowship. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript

Taxonomy of the order Mononegavirales: update 2016

In 2016, the order Mononegavirales was emended through the addition of two new families (Mymonaviridae and Sunviridae), the elevation of the paramyxoviral subfamily Pneumovirinae to family status (Pneumoviridae), the addition of five free-floating genera (Anphevirus, Arlivirus, Chengtivirus, Crustavirus, and Wastrivirus), and several other changes at the genus and species levels. This article presents the updated taxonomy of the order Mononegavirales as now accepted by the International Committee on Taxonomy of Viruses (ICTV)