Cyclic AMP plays important roles in different physiological processes, including plant defence responses. However, as little information is known on plant enzymes responsible for cAMP production/degradation, studies of cAMP functions have relied, to date, on non-specific pharmacological approaches. We therefore developed a more reliable approach, producing transgenic Arabidopsis thaliana lines overexpressing the 'cAMP-sponge' (cAS), a genetic tool that specifically buffers cAMP levels. In response to an avirulent strain of Pseudomonas syringae pv. tomato (PstAvrB), cAS plants showed a higher bacterial growth and a reduced hypersensitive cell death in comparison with wild-type (WT) plants. The low cAMP availability after pathogen infection delayed cytosolic calcium elevation, as well as hydrogen peroxide increase and induction of redox systems. The proteomic analysis, performed 24 h post-infection, indicated that a core of 49 proteins was modulated in both genotypes, while 16 and 42 proteins were uniquely modulated in WT and cAS lines, respectively. The involvement of these proteins in the impairment of defence response in cAS plants is discussed in this paper. Moreover, in silico analysis revealed that the promoter regions of the genes coding for proteins uniquely accumulating in WT plants shared the CGCG motif, a target of the calcium-calmodulinbinding transcription factor AtSR1 (Arabidopsis thaliana signal responsive1). Therefore, following pathogen perception, the low free cAMP content, altering timing and levels of defence signals, and likely acting in part through the mis-regulation of AtSR1 activity, affected the speed and strength of the immune response. Role of cAMP in plant immune response 597Generation of Arabidopsis cAS-mCherry plants expressing the NES-YC3.6 probeThe Arabidopsis Col-0 cAS-transgenic lines were crossed with the Col-0 pUBQ10-NES-YC3.6 line reported in Krebs et al. (2012). Seeds from cross-pollinated flowers were surface sterilized by vapour-
This study aimed to evaluate the biocontrol potential of bacteria isolated from different plant species and soils. The production of compounds related to phytopathogen biocontrol and/or promotion of plant growth in bacterial isolates was evaluated by measuring the production of antimicrobial compounds (ammonia and antibiosis) and hydrolytic enzymes (amylases, lipases, proteases, and chitinases) and phosphate solubilization. Of the 1219 bacterial isolates, 92% produced one or more of the eight compounds evaluated, but only 1% of the isolates produced all the compounds. Proteolytic activity was most frequently observed among the bacterial isolates. Among the compounds which often determine the success of biocontrol, 43% produced compounds which inhibit mycelial growth of Monilinia fructicola, but only 11% hydrolyzed chitin. Bacteria from different plant species (rhizosphere or phylloplane) exhibited differences in the ability to produce the compounds evaluated. Most bacterial isolates with biocontrol potential were isolated from rhizospheric soil. The most efficient bacteria (producing at least five compounds related to phytopathogen biocontrol and/or plant growth), 86 in total, were evaluated for their biocontrol potential by observing their ability to kill juvenile Mesocriconema xenoplax. Thus, we clearly observed that bacteria that produced more compounds related to phytopathogen biocontrol and/or plant growth had a higher efficacy for nematode biocontrol, which validated the selection strategy used.
INTRODUÇÃODentre as principais medidas preconizadas para o controle de doenças na cultura do arroz estão o uso de cultivares resistentes e a aplicação de fungicidas (Nunes et al., 2004). A resistência genética é uma medida altamente desejável devido ao baixo custo e à alta eficiência. No entanto, cultivares de arroz, com níveis aceitáveis de resistência, muitas vezes, não estão disponíveis para comercialização ou têm sua vida útil reduzida devido à ocorrência da quebra de resistência em virtude do surgimento de novas raças de patógenos (Cornélio et al., 2003). Existem fungicidas RESUMOO potencial dos isolados DFs185 (Pseudomonas synxatha), DFs223 (Pseudomonas fluorescens), DFs306 (não identificado), DFs416, DFs418 e DFs419 (Bacillus sp.), DFs422 (Bacillus subtilis) e DFs471 (Stenotrophomonas maltophilia) foi avaliado para o controle da mancha-parda (Bipolaris oryzae) e da escaldadura (Gerlachia oryzae) do arroz. Para tanto, sementes da cultivar El Passo L144 foram imersas em suspensão (10 7 a 10 8 UFC/mL) de cada um dos isolados, agitadas por 30 min a 10°C e semeadas em vasos, sendo utilizado o delineamento inteiramente casualizado com quatro repetições. Foram utilizadas duas testemunhas: uma tratada com solução salina e outra com solução salina mais fungicida (Carboxin+Thiran). Realizaram-se dois ensaios para cada um dos patógenos, inoculados nas plantas por aspersão, e avaliados quanto à severidade aos 7, 14 e 21 dias após a inoculação calculando a área abaixo da curva de progresso da doença. Para mancha-parda, no primeiro ensaio, o melhor desempenho foi proporcionado pelos isolados DFs185, DFs223 e DFs306 que apresentaram 80, 86 e 70% de redução da severidade. No segundo ensaio, o isolado DFs306 foi o mais eficiente deles, com redução na severidade da doença chegando a 74% em relação à testemunha, sendo que os isolados DFs223, DFs185 e DFs306 proporcionaram incremento na massa de grãos em 74, 70 e 59%, respectivamente. Para o controle da escaldadura, no primeiro ensaio, os isolados que apresentaram melhores resultados foram DFs416 e DFs418 com 65 e 59% de controle, respectivamente. No segundo ensaio, se destacaram os isolados DFs416 e DFs185 reduzindo a severidade da doença em 64 e 60%, respectivamente sem incrementar a produção. Palavras-chave: Bipolaris oryzae, Gerlachia oryzae, Oryza sativa, controle biológico. ABSTRACT Seed microbiolization for the control of rice brown spot and leaf scaldThe potential of isolates DFs185 (Pseudomonas synxatha), DFs223 (Pseudomonas fluorescens), DFs306 (unidentified), DFs416, DFs418 and DFs419 (Bacillus sp.), DFs422 (Bacillus subtilis) and DFs471 (Stenotrophomonas maltophilia) was evaluated for the control of rice brown spot (Bipolaris oryzae) and leaf scald (Gerlachia oryzae). Rice seeds cv. El Passo L144 were immersed in a suspension (10 7 a 10 8 CFU/mL) of each of the isolates, agitated for 30 min at 10°C and sowed in pots, in complete randomized design with four replications. Two controls were used: one with seeds treated with saline solution, and the other with saline...
Emphasis has been given on selection of micro-organism for biological control. However, in order to evaluate the biological control potential of a great number of micro-organisms in a small period of time it is necessary to develop an efficient bioassay. Seven hundred and sixty bacterial isolates from different habitats, were selected for compatibility with Rhizobium leguminosarum bv. phaseoli (SEMIA 4077 e SEMIA 4080). Among them 596 isolates were ineffective against both rhizobia. Bean seeds immersed in suspension of each one of these isolates were agitated for 5 hours at 10 o C and sowed in non-sterilized soil. The plants were kept in greenhouse. After the development of cotyledonary and primary leaves, these were removed and bioassayed for Xanthomonas axonopodis pv. phaseoli (XAP) control. In the cotyledonary leaves, it was observed that the isolate DFs093 offered 100% control, DFs041 and DFs1297 offered 90% and DFs490, DFs769, DFs831, DFs842 and DFs843 offered 80% control. In the primary leaves, the DFs482 isolated offered 100% and the DFs080, DFs348, DFs513, DFs622, DFs769, DFs842 and DFs912 offered 80% of XAP control.
Potential of microbiolization of rice seeds with rhizobacteria for root-knot nematode biocontrolThe potential of eight rhizobacteria that effectively control fungal diseases in rice [DFs185 (Pseudomonas synxantha), DFs223 (P. fluorescens), DFs306 (not identified), DFs416, DFs418 e DFs419 (Bacillus sp.), DFs422 (Bacillus subtilis), DFs471 (Stenotrophomonas malthophilia)] was evaluated for the control of Meloidogyne graminicola in vitro and in irrigated rice in the greenhouse. The rhizobacteria DFs185, DFs223, DFs306, DFs416 and DFs419 exceeded in reducing hatching and in increasing the mortality of M. graminicola J2. All rhizobacteria were able to produce at least one compound associated with the biocontrol of nematodes (chitinasis, phosphatases, lipases, proteinasis and siderophores). The number of eggs and galls of M. graminicola in rice plants was reduced compared to plants from seeds not microbiolized. The nematode reproduction factor was reduced on average by 29%, highlighting rhizobacteria DFs185 and DFs223 that reduced 35%. This result is significant because the rhizobacteria also control fungal diseases.
Poly (3-hydroxybutyrate) (P(3HB)) is the most studied thermoplastic biopolymer belonging to the polyhydroxyalkanoate (PHA) family, the main features of which include rapid biodegradability and biocompatibility. The bioplastic recovery process is an important step during production and can directly influence the characteristics of PHAs. However, more efficient methods for the production of P(3HB) are necessary to make it economically viable. The aim of the present study was to improve the standard, chloroformbased, extraction step for the recovery of P(3HB). The polymer was produced using a Ralstonia solanacearum strain. The following parameters were improved in the recovery process: heating time, separation method (filtration or liquid-liquid phase separation), biomass state (fresh or dry cell concentrate) and the solvent:biomass ratio. By improving the chemical extraction of P(3HB) we recovered 98% of the cumulative polymer and reduced the heating time by 75%. Furthermore, we improved the separation process and developed an extraction solution that was faster and more economical.
RESUMOObjetivou-se avaliar o efeito de oito isolados bacterianos pré-selecionados de Pseudomonas synxatha, P. fluorescens, Bacillus subtilis, Bacillus sp. e Stenotrophomonas malthophilia no controle da queima-das-bainhas do arroz, causada por Rhizoctonia solani. Sementes da cultivar El Passo L144 foram imersas em suspensão (A 540 =0,5) de cada um dos isolados e agitadas por 30 min a 10�C. Sementes imersas somente em solução salina e em salina mais fungicida (Carboxin + Thiran) foram utilizadas como testemunhas. Foram semeadas 10 sementes por vaso, em quatro repetições, dispostas em delineamento completamente casualizado. Foram realizados três ensaios, sendo que no primeiro foi possível selecionar três isolados como promissores, com reduções na severidade da doença atingindo 50, 33,3 e 16,7 %, respectivamente. Estes isolados foram utilizados nos ensaios posteriores, instalados em casa de vegetação e conduzidos até o ponto de colheita, onde foi possível observar o efeito biocontrolador propiciado, principalmente, pelo isolado de P. fluorescens DFs223, com reduções significativas na severidade da doença chegando a 88 e 91,7% no segundo e terceiro ensaios respectivamente. Em ambos os ensaios, houve incremento tanto do número de panículas quanto do número de perfilhos e da massa seca de raízes em até 42,8, 81,2 e 113% respectivamente, nas plantas tratadas com o isolado de P. fluorescens DFs223.Palavras-chave adicionais: Rhizoctonia solani, Oryza sativa, biocontrole, rizobactérias. ABSTRACTBiological control of rice sheath blight by microbiolization of seeds with antagonist bacteria Eight pre-selected bacterial strains were evaluated for control of rice sheath blight caused by Rhizoctonia solani, Pseudomonas synxatha, P. fluorescens, Bacillus subtilis, Bacillus sp. and Stenotrophomonas malthopilia. Rice seeds cv. El Passo L144 were treated by immersion in a bacterial suspension (A 540 =0.5) of each isolate under agitation at 10�C for 30 min. Seeds immersed in saline solution and in saline plus fungicide (Carboxin + Thiran) were used as control. Ten seeds per pot were used, in four replications, in a completely randomized design. Three assays were performed, and in the first it was possible to select the three strains which reduced the severity of the disease, reaching 50, 33.3 and 16.7%, respectively. These isolates were used in the two last assays, which were maintained in greenhouse until the harvesting stage, where it was possible to observe the biocontrol effect, mainly by the strain of P. fluorescens DFs223, with significant reduction in the disease severity, reaching 88 and 91.7% in the second and third assays, respectively. There was an increase in the number of panicles, as well as in the number of tillers and dry mass of roots, reaching to 42.8, 81.2 and 113 % respectively, in plants treated with this isolate in both assays.Additional keywords: Rhizoctonia solani, Oryza sativa, biocontrol, rhizobacteria. INTRODUÇÃOA cultura do arroz irrigado (Oryza sativa L.) está sujeita ao ataque de várias doenças. Em quase...
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