CHARACTERIZATION of RALSTONIA SOLANACEARUM STRAINS FROM BRAZIL USING MOLECULAR METHODS and PATHOGENICITY TESTS

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Avaliação da agressividade e caracterização genética de linhagens de Ralstonia Solanacearum isoladas de diferentes plantas hospedeiras

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Enxertia no controle da murcha bacteriana na atividade de enzimas e produção em tomateiro

Pós-graduação em Agronomia (Horticultura) - FCA

Resposta enzimática, fisiológica e produtiva do tomateiro e desempenho de porta enxertos resistentes à murcha bacteriana

Pós-graduação em Agronomia (Horticultura) - FCA

Risposte di resistenza a batteri fitopatogeni di importanti specie coltivate indotte da molecole segnale di diversa natura

A modern management of crop protection should be based on integrated control programmes, including the use of environmentally safe products. Antagonistic/beneficial bacteria and resistance inducers may have a great potential in the prophylaxis of diseases caused by common and quarantine pathogens. This work was carried out to confirm the ability of the known strain IPV-BO G19 (Pseudomonas fluorescens) against fire blight (Erwinia amylovora), as well as to evaluate their efficacy against southern bacterial wilt of tomato (Ralstonia solanacearum) and grapevine crown gall (Agrobacterium vitis). A virulent strain of R. solanacearum race 3 was inhibited by the antagonist on plate. When the pathogen was inoculated 48 h after their application to the root apparatus of tomato plants grown in a climatic chamber, bacterial wilt progression rate was clearly reduced. Moreover the defence response evoked by IPV-BO G19 was studied in tomato plants by monitoring the transcription of genes codifying for three PRs as PR-1a, PR-4, PR-5 and for an intracellular chitinase using multiplex RT-PCR and Real Time RT-PCR. In two field trials during 2005 and 2006, the strain IPV-BO G19 was compared with biofungicides and some abiotic elicitors to protect actively growing shoots of pear scions against fire blight. In both trials, IPV-BO G19 plus Na-alginate gave a high level of protection, three weeks after wound inoculation with E. amylovora. In pear leaf tissues treated with the antagonistic strain IPV-BO G19, catalase, superoxyde dismutase and peroxidise activity was evaluated as markers of induced resistance. The IPV-BO G19 strain was compared with other bioagents and resistance inducers to prevent grapevine crown gall under glasshouse and vineyard conditions.

Organic Practices for the Production of Muskmelon

Cucurbit crops, especially muskmelon and cucumber, attract cucumber beetles, which vector bacterial wilt: Erwinia tracheiphila, causing significant crop losses. High beetle densities are associated with high bacterial wilt incidence, which usually occurs during the first stages of plant establishment. Growers of organic muskmelon need effective ways to manage the cucumber beetle/bacterial wilt complex.

Detección de Ralstonia solanacearum en Solanum tuberosum L. en el Estado de Sonora, México

Ralstonia solanacearum (Rs) produce la enfermedad cuarentenaria denominada marchitez bacteriana en papa. México es un país importador de semilla de Estados Unidos de América y Canadá, aspecto significante para provocar una eventual introducción de esta enfermedad en áreas con amplias extensiones de papa. Sonora es una región importante en relación con la producción de este cultivo. Por lo anteriormente expuesto, se realizó la presente investigación, teniendo como objetivos: a) la producción de antisuero para la bacteria Rs; b) diagnosticar Rs en tubérculos de importación que se utilizan para siembra, y en tubérculos de procedencia mexicana para consumo humano, que son utilizados como semilla; c) la detección de la bacteria durante el desarrollo vegetativo de lotes de papa en Sonora, México. Se analizó tubérculo semilla de importación, de consumo humano, plantas de papa, hojas y tubérculos de producción; los métodos de detección utilizados fueron medios de cultivos específicos, ELISA, antisuero producido y pruebas de patogenicidad. Los resultados mostraron positiva la presencia de Rs en tubérculos de consumo; en tubérculos de importación y en etapas vegetativas fue negativa. Cada prueba de detección por separado no debe ser utilizada como método único; la presencia de Rs representa un riesgo de eventual manifestación de la enfermedad, por lo que es necesario que las áreas productoras realicen actividades de control preventivo fitosanitario.

Identification of the causal agent of pistachio dieback in Australia

Symptoms associated with pistachio dieback in Australia include decline (little or no current season growth), xylem staining in shoots two or more years old, trunk mu and limb lesions (often covered by black, superficial fungal growth), excessive exudation of resin, dieback and death of the tree. Bacteria belonging to the genus Xanthomonas have been suggested as the causal agent. To confirm the constant association between these bacteria and the disease syndrome, the absence of other pathogens and the identity of the pathogen, we performed a series of isolations and pathogenicity tests. The only microorganism consistently isolated from diseased tissue was a bacterium that produced yellow, mucoid colonies and displayed morphological and cultural characteristics typical of the genus Xanthomonas. Database comparisons of the fatty acid and whole-cell protein profiles of five representative pistachio isolates indicated that they all belonged to X. translucens, but it was not possible to allocate the isolates to pathovar. Pathogenicity tests on cereals and grasses supported this identification. However, Koch's postulates have been only partially fulfilled because not all symptoms associated with pistachio dieback were reproduced on inoculated two-year-old pistachio trees. While discolouration was observed, dieback, excessive resinous exudate and trunk and limb lesions were not produced; expression of these symptoms may be delayed, and long-term monitoring of a small number of inoculated trees is in progress.

Pseudomonas fluorescentes provenientes da rizosfera de solanáceas no controle de Ralstonia solanacearum em tomateiro

Dissertação (mestrado)—Universidade de Brasília, Instituto de Ciências Biológicas, Departamento de Fitopatologia, Programa de Pós-Graduação em Fitopatologia, 2015.

Formulation and delivery of the bacterial antagonist Bacillus subtilis for management of lentil vascular wilt caused by Fusarium oxysporum f. sp lentis

Different formulations of Bacillus subtilis were prepared using standard laboratory protocols. Bacillus subtilis survived in glucose and talc powders at 8.6 and 7.8 log(10) CFU/g, respectively, for 1 year of storage at room temperature compared with 3.5 log(10) CFU/g on a peat formulation. Glasshouse experiments using soil and seed treatments were conducted to test the efficacy of B. subtilis for protecting lentil against the wilt disease caused by Fusariumoxysporum f. sp. lentis. Seed treatments with formulations of B. subtilis on glucose, talc and peat significantly enhanced its biocontrol activity against Fusarium compared with a treatment in which spores were applied directly to seed. The formulations decreased disease severity by reducing colonization of plants by the pathogen, promoting their growth and increased the dry weight of lentil plants. Of these treatments the glucose and talc-based powder formulations were more effective than the peat formulation and the spore application without a carrier. It was shown that the B. subtilis spores applied with glucose were viable for longer than those applied with other carriers. Seed treatment with these formulated spores is an effective delivery system that can provide a conducive environment for B. subtilis to suppress vascular wilt disease on lentil and has the potential for utilization in commercial field application.

Antagonistic potential of bacterial isolates associated with entomopathogenic nematodes against tomato wilt caused by Fusarium oxysporum f.sp., Lycopersici under greenhouse conditions

Three concentrations of Xenorhabdus nematophila and Xenorhabdus spp., (4x10(5,) 4x10(6,) 4x10(7) cells/ml) were evaluated in the laboratory and in pot experiments to test their antagonistic effects on Fusarium oxysporum f.sp., lycopersici. All concentrations effectively inhibited its growth on agar plates. In soil under greenhouse conditions treatments with each bacterium at 4x10(7) cells/ml reduced the disease incidence of tomato by up to 40.38 and 47.54% respectively and there were significant increases of plant biomass by 198 and 211% respectively. The rhizosphere population of Fusarium oxysporum f.sp., lycopersici was reduced by 97%. The Xenorhabdus spp., was comparatively more effective than X. nematophila.

Evaluation of pyraclostrobin and acibenzolar-S-methyl on common bacterial blight of snap bean

Assays were done under greenhouse conditions in order to evaluate the effect of pyraclostrobin (0.0375, 0.0750 and 0.150 mL.L-1) and acibenzolar-S-methyl (ASM) (0.025 g.L-1) in common bacterial blight on leaves of snap beans cultivar Braganca. These chemicals were sprayed at three different times: five days before; five days before + five days after; and five days after leaf inoculation with an isolate of Xanthomonas axonopodis pv. phaseoli. They were determinate the levels of polyphenoloxidase, peroxidase and total soluble proteins on inoculated and non-inoculated leaves of snap beans sprayed with pyraclostrobin (0.075 g.L-1) and ASM (0.025 g.L-1). All concentration of pyraclostrobin and ASM reduced the area under the disease progress curve (AUDPC) on leaves of snap beans, and the least AUDPC value was observed when this products were sprayed five days before + five days after inoculation. Higher levels of polyphenoloxidase, peroxidase and the total soluble proteins were observed on leaves sprayed with pyraclostrobin or ASM.

Resistance to Fusarium wilt in common bean cultivars and lines in pre-commercial stage.

The objective of this study was to identify common bean cultivars with resistance to Fusarium wilt.

Bacterial community associated to the pine wilt disease insect vectors Monochamus galloprovincialis and Monochamus alternatus

Monochamus beetles are the dispersing vectors of the nematode Bursaphelenchus xylophilus, the causative agent of pine wilt disease (PWD). PWD inflicts significant damages in Eurasian pine forests. Symbiotic microorganisms have a large influence in insect survival. The aim of this study was to characterize the bacterial community associated to PWD vectors in Europe and East Asia using a culture-independent approach. Twenty-three Monochamus galloprovincialiswere collected in Portugal (two different locations); twelve Monochamus alternatus were collected in Japan. DNA was extracted from the insects’ tracheas for 16S rDNA analysis through denaturing gradient gel electrophoresis and barcoded pyrosequencing. Enterobacteriales, Pseudomonadales, Vibrionales and Oceanospirilales were present in all samples. Enterobacteriaceae was represented by 52.2% of the total number of reads. Twenty-three OTUs were present in all locations. Significant differences existed between the microbiomes of the two insect species while for M. galloprovincialis there were no significant differences between samples from different Portuguese locations. This study presents a detailed description of the bacterial community colonizing the Monochamus insects’ tracheas. Several of the identified bacterial groups were described previously in association with pine trees and B. xylophilus, and their previously described functions suggest that they may play a relevant role in PWD.

Survey of Viruses Affecting Faba Bean in Six Arab Countries

A field survey of faba bean (Vicia [aba L.) for viruses in six Arab countries showed the presence of nine viruses. Bean leaf roll virus (BLRV), bean yellow mosaic virus (BYMV), broad bean mottle virus (BBMV) and to a lesser extent broad bean stain virus (BBSV) were the most common. When testing with ELISA 789 samples with symptoms suggestive of virus infection collected from Egypt, Lebanon, Morocco, Sudan, Syria and Tunisia, BBMV was detected in 203 samples, BBSV in 151, broad bean true mosaic virus (BBTMV) in 7, broad bean wilt virus (BBWV) in 47, BYMV in 314, cucumber mosaic virus (CMV) in 96, pea enation mosaic virus (PEMV) in 31, and pea seed-borne mosaic virus (PSbMV) in 49 samples. Identity of selected field isolates was confirmed by electron microscopy and host reaction studies. In a yield experiment, infection with BYMV, BBMV and BBSV 11 weeks after sowing (pre-flowering) led to 81, 54 and 84% yield loss, respectively. Inoculation with the same viruses 15 weeks after sowing (flowering) and 20 weeks after sowing (pod setting) led to 56, 84 and 18%, and 39, 37 and 18% yield loss, respectively.