RESUMO: O objetivo deste estudo foi identificar molecularmente diferentes espécies de Aeromonas isoladas de tambaqui (Colossoma macropomum) de cultivo do norte do Brasil e avaliar seu potencial patogênico pela presença de genes de virulência. A partir da extração do DNA bacteriano, foi realizada a PCR (reação em cadeia da polimerase) dos primers 16S rDNA, aerA (enterotoxina citolítica), ast (enterotoxina citotóxica) e act (enterotoxina citotóxica). Dos 24 isolados avaliados, oito amplificaram o gene ast, um amplificou o gene act, mas o gene areA não foi amplificado em nenhum isolado. O sequenciamento do primer 16S rRNA revelou uma predominância da espécie Aeromonas jandaei (92%). Aeromonas taiwanensis (4%), pela primeira vez isolada em peixes no Brasil, e Aeromonas hydrophila (4%) apareceram cada uma como apenas um isolado. Os resultados mostram que 32% das Aeromonas isoladas de tambaqui de cultivo apresentam considerável potencial patogênico para danos sistêmicos, uma vez que os primers de PCR selecionados estão codificando os genes de virulência mais comuns em Aeromonas com alta intensidade patogênica.

ABSTRACT: The aim of this study was to molecularly identify different species of Aeromonas isolated from farmed tambaqui (Colossoma macropomum) from North Brazil, and evaluate their pathogenic potential by the presence of virulence genes. From the extraction of bacterial DNA, PCR (polymerase chain reaction) of the primers 16S rDNA, aerA (cytolytic enterotoxin), ast (cytotoxic enterotoxin) and act (cytotoxic enterotoxin) were performed. Of 24 isolates evaluated, eight amplified the ast gene, one amplified the act gene, but the areA gene was not amplified in any isolate. Sequencing of the 16S rRNA primer revealed a predominance of Aeromonas jandaei specie (92%). Aeromonas taiwanensis (4%), for the first time isolated from fish in Brazil, and Aeromonas hydrophila (4%) each appeared as just one isolate. Results showed that 32% of Aeromonas isolated from farmed tambaqui have considerable pathogenic potential for systemic damage, since the selected PCR primers are encoding the most common virulence genes in Aeromonas with high pathogenic intensity.

RESUMO O uso de bactérias na promoção do crescimento e no controle biológico de doenças em plantas pode minimizar a contaminação ambiental causada pela aplicação indiscriminada de pesticidas e fertilizantes químicos. Objetivamos avaliar a promoção do crescimento e o controle biológico de Corynespora cassiicola em mudas de tomate mediadas por bactérias benéficas isoladas de solo amazônico não-rizosférico contendo diferentes dosagens de biocarvão, e identificar a quais grupos de bactérias os isolados pertencem. Obtivemos 200 isolados de parcelas experimentais contendo doses de biocarvão de 0, 40, 80 e 120 t ha-1. Destes, 53 foram selecionados por testes de colonização radicular. Com base nos parâmetros de promoção do crescimento, 25 isolados foram selecionados, identificados através de análise molecular e avaliados para produção de ácido indolacético (AIA), solubilização de fosfato e controle biológico. A melhor dose de biocarvão para a formação de colônias foi 40 t ha-1, e um modelo de regressão indicou 34 t ha-1como dose ótima. A produção de AIA foi observada em 18 (75%) isolados e dois (8%) isolados foram capazes de solubilizar fosfato. A eficiência na promoção do crescimento das raízes foi de até 125%, e a porcentagem de proteção das plantas variou de 50,3 a 59,0%. A caracterização molecular indicou que as bactérias utilizadas nesse estudo pertencem aos gêneros Bacillus e Lysinibacillus.

ABSTRACT The use of bacteria in growth promotion and biological control of plant diseases can minimize environmental contamination caused by the indiscriminate use of pesticides and chemical fertilizers. We aimed to evaluate growth promotion and biological control of Corynespora cassiicola in tomato seedlings mediated by beneficial bacteria isolated from a non-rhizospheric Amazon soil containing different amounts of biochar, and to identify to which groups of bacteria the strains belong. We obtained 200 strains of bacteria from experimental plots containing biochar doses of 0, 40, 80 and 120 t ha-1. Of these, 53 strains were selected by root colonization tests. Based on growth promotion parameters, 25 strains were screened, identified by molecular characterization and evaluated for indoleacetic acid (IAA) production, phosphate solubilization and biological control. The best dose of biochar for colony formation was 40 t ha-1, and a regression model indicated 34 t ha-1 as the optimal dose. The production of IAA was observed in 18 (75%) strains, and two (8%) strains were able to solubilize phosphate. The efficiency in root growth promotion was up to 125%, and the percentage of plant protection ranged from 50 to 59%. Molecular characterization showed that the bacteria used in this study belong to the genera Bacillus and Lysinibacillus.

Black Sigatoka caused by Mycosphaerella fijiensis is a foliar disease that affects banana plants and large amounts of fungicides are required to prevent crop losses. Intensive applications of single-site fungicides can select for fungicide-resistant isolates. The objective of this study was to assess the sensitivity of 60 isolates of M. fijiensis to commonly used fungicides. Using two different protocols, microtiter and Petri plate tests, the effective concentration at which mycelium growth is reduced by 50% (EC50) was determined for thiophanatemethyl, tebuconazole, chlorothalonil and mancozeb. Additionally, partial sequences of the cytochrome b gene were obtained for 46 isolates to detect the G143A mutation, commonly associated with strobilurin resistance. The EC50 values for tebuconazole and thiophanate-methyl ranged from 0.02 to 1.39 and from 0.008 to 8.22 µg mL-1, respectively. For chlorothalonil, the lowest and highest EC50 values were 0.39 µg mL-1 and 53.7 µg mL-1, respectively. For mancozeb, approximately 50% of the isolates had EC50 values greater than 1000 µg mL-1. No mutation was found in the isolates assayed for strobilurin resistance. There was no correlation between sensitivity levels to any fungicide and geographic region. Low EC50 values were estimated for most fungicides but, some isolates had high EC50 values for mancozeb.

The objective of this study was to identify RAPD and SSR markers associated with a resistant allele for angular leaf spot (Phaeoisariopsis griseola) from the line 'ESAL 550', derived from the Andean 'Jalo EEP 558' cultivar, to assist selection of resistant genotypes. The resistant line 'ESAL 550' and the susceptible cultivar 'Carioca MG' were crossed to generate F1 and F2 populations. One hundred and twenty F2:3 families were evaluated. The DNA of the 12 most resistant families was bulked and the same was done with the DNA of the 10 most susceptible, generating two contrasting bulks. One RAPD and one SSR marker was found to be linked in coupling phase to the resistant allele. The SSR marker was amplified by the primer PV-atct001(282C), and its distance from the resistant allele was 7.6 cM. This is the most useful marker for indirect selection of resistant plants in segregating populations. The RAPD marker was amplified by the primer OPP07(857C) linked in coupling phase to the resistant allele, and distant 24.4 cM. Therefore, this RAPD marker is not so useful in assisting selection because it is too far from the resistant allele.