The role of melanin in the grapevine trunk disease pathogen Lasiodiplodia gilanensis

Lasiodiplodia (Botryosphaeriaceae) includes fungi that are considered among the most aggressive to grapevine, capable of causing cankers and necrotic lesions which eventually lead to death of host plants. A common characteristic of this genus is the presence of melanin in conidia and mycelium. Melanin is produced by the oxidation of phenolic and/or indolic compounds. For some fungi, this pigment is an essential factor for pathogenicity. This study characterized the types and the roles of melanin produced by Lasiodiplodia gilanensis. Using specific melanin inhibitors, L. gilanensis was shown to synthesize DOPA-melanin, DHN-melanin, and pyomelanin. DOPA-melanin was shown to be involved in production of aerial mycelium and protection against enzymatic lysis and oxidative stress; DHN-melanin to be involved in ramification of mycelium when exposed to nutrient deficiency; and pyomelanin to be related with hyphae development. The fungus used tyrosine as a precursor of DOPAmelanin and as carbon and nitrogen sources, and produced melanin inside the piths of infected plants. Genes involved in melanin synthesis were conserved among the Botryosphaeriaceae, highlighting the importance of melanin in this family.EEA MendozaFil: Rangel Montoya, Edelweiss Airam. Centro de Investigación Científica y de Educación Superior de Ensenada (CICESE). Departamento de Microbiología; MéxicoFil: Paolinelli, Marcos. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Mendoza; ArgentinaFil: Paolinelli, Marcos. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina; ArgentinaFil: Rolshausen, Philippe. University of California Riverside. Department of Botany and Plant Sciences; Estados UnidosFil: Hernandez Martinez, Rufina. Centro de Investigación Científica y de Educación Superior de Ensenada (CICESE). Departamento de Microbiología; Méxic

Characterization of Lasiodiplodia species associated with grapevines in Mexico

Botryosphaeria dieback is one of the most prevalent grapevine trunk diseases (GTDs), and is caused by fungi in the Botryosphaeriaceae. Fungi invade grapevine vascular systems mainly through pruning wounds, and cause cankers and necrotic lesions, which lead to grapevine decline and death. Lasiodiplodia theobromae has been reported as a highly virulent pathogen of grapevine, and was previously reported in Mexican vineyards. The taxonomy of Lasiodiplodia was recently revised, adding new species, and some were reduced to synonymy. This study aimed to characterize Lasiodiplodia producing grapevine dieback symptoms in Sonora and Baja California, Mexico. Using the phylogenetic markers tef1-α and ITS regions, Lasiodiplodia brasiliensis, L. crassispora, L. exigua, and L. gilanensis were identified. Lasidiplodia exigua was the most prevalent species. Lasiodiplodia brasiliensis and L. gilanensis were very virulent to ‘Cabernet Sauvignon’ plants, while L. exigua and L. gilanensis were less virulent, and L. crassispora did not produce lesions at 2 months post-inoculation. The optimum temperature of the Lasiodiplodia spp. was 28°C, but all four species grew up to 37°C, and the isolates of L. exigua grew slowly at 40°C. This is the first report of the four of Lasiodiplodia species in vineyards of Mexico.EEA MendozaFil: Rangel Montoya, Edelweiss A. Centro de Investigación Científica y de Educación Superior de Ensenada (CICESE). Departamento de Microbiología; MéxicoFil: Paolinelli, Marcos. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Paolinelli, Marcos. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Mendoza; ArgentinaFil: Rolshausen, Philippe E. University of California, Riverside. Department of Botany and Plant Sciences; Estados UnidosFil: Valenzuela Solano, Cesar. Instituto Nacional de Investigaciones Forestales, Agrícolas y Pecuarias (INIFAP). Campo Experimental Costa de Ensenada; MéxicoFil.: Hernandez Martinez, Rufina. Centro de Investigación Científica y de Educación Superior de Ensenada (CICESE). Departamento de Microbiología; Méxic

Bioprospecting arsenite oxidizing bacteria in the soil of the Comarca Lagunera

Resumen El arsénico es uno de los metaloides más tóxicos presente en el ambiente y la exposición prolongada a este metal causa efectos crónicos en la salud. Por ello, la búsqueda de alternativas amigables con el medio ambiente, para el tratamiento de agua y suelos contaminados con arsénico es importante. En este trabajo se aislaron cepas bacterianas de suelos con presencia de arsénico en la Comarca Lagunera, para analizar aquellas con capacidad oxidante de arsenito. Las cepas 04-SP1qa y 14-SP1qh de metabolismo quimiolitoautotrófico y quimioheterotrófico, respectivamente, tuvieron mayor actividad de la enzima arsenito oxidasa. Las condiciones óptimas de crecimiento y la actividad enzimática de dichas cepas se investigaron. La cepa 04-SP1qa presentó actividad enzimática específica de 0.162 μmol·min- 1 ·mg-1, constante de Michaelis-Menten (Km) de 3.37 μM y velocidad máxima (Vmax) de 5.20 μM·min-1·mg-1 en condiciones óptimas de pH 8.0 y 40 °C. La cepa 14-SP1qh presentó actividad enzimática específica de 0.16 μmol·min-1·mg-1, Km de 3.70 μM y Vmax de 14.39 μM·min-1·mg-1 a pH 7.0 y 40 °C. Los resultados demostraron la presencia de bacterias oxidantes de arsenito con actividad enzimática en suelos de la Comarca Lagunera, identificando potencial para desarrollar nuevas tecnologías de biorremediación de aguas y suelos contaminados con arsénico en la región

Lasiodiplodia gilanensis used as model for understanding the pathogenicity of Botryosphaeriaceae

Lasiodiplodia spp. cause degenerative diseases, dieback and plant death in a wide range of woody plants. To understand the fungal behavior of the genus we are using Lasiodiplodia gilanensis as a model. A transcriptional study revealed that the fungus is capable of using the phenylpropanoid precursors and salicylic acid to avoid the host defense response of the plant. On another hand, several genes encoding enzymes involved in different melanin synthesis pathways; 3,4-dihydroxyphenylalanine (DOPA)-melanin, 1,8-dihydroxynaphthalene (DHN)-melanin and pyomelanin; were identified and their production evauated, concluding that the fungus use diferent types of melanin to overcome environmental stress. An in silico analysis shows the presence of those genes in all the available genomes of Botryosphaeriaceae in the GeneBank, evidencing the importance of the melanin in this family. L. gilanensis produces siderophores of catechol and hydroxamate-type as well as naturally esterified fatty acids that might have a role in plant growth regulation. A plethora of hydrolytic enzymes is also produced, including xylanases, ligninases, cellulases, pectinases, cutinases, and hemicelluloses. Another secondary metabolite produced is oxalic acid, reported as a pathogenicity factor in other fungi, which role is under evaluation. Microscopical observations showed that the fungus uses the starch deposited in the ray cells as carbon source, induces the production of suberin and phenolic compounds and colonizes the vascular cambium, ray parenchyma, and the vascular bundles. These studies extend our understanding of the pathogenicity of a widely distributed pathogenic fungus with our final goal to control the diseases it causes.Fil: Rangel Montoya Edelweiss. Consejo Nacional de Ciencia y Tecnología de México. Centro de Investigación Científica y de Educación Superior de Ensenada Baja California; MéxicoFil: Paolinelli, Marcos. Instituto Nacional de Tecnología Agropecuaria. Centro Regional Mendoza-San Juan. Estación Experimental Agropecuaria Mendoza; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Hernández Martínez Rufina. Consejo Nacional de Ciencia y Tecnología de México. Centro de Investigación Científica y de Educación Superior de Ensenada Baja California; México11th International Workshop on grapevine trunk diseasesPentictonCanadáOregon Wine Research Institut

Biocontrol of <i>Macrophomina phaseolina</i> Using <i>Bacillus amyloliquefaciens</i> Strains in Cowpea (<i>Vigna unguiculata</i> L.)

The fungus Macrophomina phaseolina is the causal agent of charcoal rot in many crops, such as strawberries and beans. Symptoms include stem and root rot and chlorotic foliage. This disease’s management is complicated because the pathogen forms resistant microsclerotia. This work aimed to obtain bacterial isolates for the biocontrol of M. phaseolina in arid regions. Two strains that grew well under low pH and high salinity, named BsA3MX and BsC11MX, were isolated and identified as B. amyloliquefaciens, based on their morphology and analysis of the 16S ribosomal RNA. Both strains inhibited M. phaseolina up to 66.8% in vitro through the combined action of volatile and diffusible compounds. Furthermore, they produce siderophores and indole-3-acetic acid (IAA), have ACC-deaminase activity, solubilize phosphate and zinc, and decrease microsclerotia germination. Moreover, in greenhouse assays using cowpea plants (Vigna unguiculata L.), strain BsA3MX reduced lesions caused by M. phaseolina and induced a significant increase in foliage and root biomass. Overall, these results suggest B. amyloliquefaciens BsA3MX and BsC11MX can be used as biological control agents against M. phaseolina in arid zones

IMPACTO DEL ARSÉNICO EN EL AMBIENTE Y SU TRANSFORMACIÓN POR MICROORGANISMOS

El arsénico (As) es uno de los metaloides más tóxicos presentes en el medio ambiente y la especiación de éste depende de diversos factores químicos, físicos y biológicos. La distribución y contaminación del arsénico se debe a procesos naturales y antropogénicos, y su problemática se debe a su fácil movilización en el ambiente. Las altas concentraciones de arsénico en agua y suelo se han convertido en un problema global, ya que las exposiciones prolongadas a este metaloide pueden causar daños crónicos a la salud. Dicha situación es particularmente importante en la Comarca Lagunera en México. Por lo tanto, se ha vuelto necesaria la búsqueda de nuevas estrategias para la remoción de este metaloide, entre las cuales, la biorremediación microbiana ha cobrado importancia como una alternativa amigable con el ambiente en la solución de este problema. Los microorganismos juegan un papel fundamental en la especiación del arsénico, ya que un gran número de estos tiene la capacidad de transformar el arsénico a pesar de su toxicidad puesto que han desarrollado diferentes mecanismos que les permiten utilizar el arsénico en su metabolismo, ya sea en su forma reducida de arsenito, o en su forma oxidada de arseniato mediante reacciones de óxido-reducción transformación enzimática, metilación, quelación, exclusión e inmovilización. Este trabajo, trata sobre el impacto que tiene la especiación de arsénico, especialmente del arsenito (As III), en el medio ambiente, el riesgo que representa la contaminación para la salud, así como los mecanismos bioquímicos microbianos para la oxidación del arsenito, disminuir su toxicidad y el desarrollo de estrategias limpias y amigables con el medio ambiente para la remoción de arsénico de aguas y suelos contaminados