Andréa Bittencourt Moura

Bioassay for selection of biocontroller bacteria against bean common blight (Xanthomonas axonopodis pv. phaseoli)

Tem-se dado muita enfase ao controle biologico mediante selecao de microorganismos. Porem, para se avaliar o potencial de biocontroladores de forma massal e em pequeno intervalo de tempo e necessario desenvolver um bioensaio eficiente. Bacterias de diferentes sitios, num total de 760 isolados, foram selecionadas para compatibilidade com Rhizobium leguminosarum bv. phaseoli estirpes SEMIA 4077 e SEMIA 4080, onde 596 isolados foram inefetivos contra ambos rizobios. Sementes de feijao foram imersas em suspensao de cada um destes isolados sendo agitadas por 5 horas a 10oC, plantadas em solo nao esterelizado, sendo as plantas mantidas em casa de vegetacao. Apos o desenvolvimento das folhas cotiledonares e folhas primarias, estas foram retiradas e avaliadas por bioensaio para o controle de Xanthomonas axonopodis pv. phaseoli (XAP). Nas folhas cotiledonares, observou-se que o isolado DFs093, proporcionou 100% de controle, DFs041 e DFs1297 propiciaram, 90% e DFs490, DFs769, DFs831, DFs842 e DFs843 proporcionaram 80% de controle. Nas folhas primarias, o isolado, DFs482 propiciou 100% e os isolados DFs080, DFs348, DFs513, DFs622, DFs769, DFs842 e DFs912 proporcionaram 80% de controle para XAP.

Microbiolização de sementes para o controle da mancha-parda e da escaldadura em arroz irrigado

O potencial dos isolados DFs185 (Pseudomonas synxatha), DFs223 (Pseudomonas fluorescens), DFs306 (nao 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 suspensao (107 a 108 UFC/mL) de cada um dos isolados, agitadas por 30 min a 10oC e semeadas em vasos, sendo utilizado o delineamento inteiramente casualizado com quatro repeticoes. Foram utilizadas duas testemunhas: uma tratada com solucao salina e outra com solucao salina mais fungicida (Carboxin+Thiran). Realizaram-se dois ensaios para cada um dos patogenos, inoculados nas plantas por aspersao, e avaliados quanto a severidade aos 7, 14 e 21 dias apos a inoculacao calculando a area abaixo da curva de progresso da doenca. Para mancha-parda, no primeiro ensaio, o melhor desempenho foi proporcionado pelos isolados DFs185, DFs223 e DFs306 que apresentaram 80, 86 e 70% de reducao da severidade. No segundo ensaio, o isolado DFs306 foi o mais eficiente deles, com reducao na severidade da doenca chegando a 74% em relacao a testemunha, sendo que os isolados DFs223, DFs185 e DFs306 proporcionaram incremento na massa de graos 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 doenca em 64 e 60%, respectivamente sem incrementar a producao.

Potencial da microbiolização de sementes de arroz com rizobactérias para o biocontrole do nematoide das galhas

The 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.

Influência da microbiolização de sementes de feijão sobre a transmissão de Colletrichum Lindemuthianum (Saac e Magn)

Isolates of Pseudomonas veronii (DFs513), Bacillus spp. (DFs093 and DFs348), Bacillus cereus (DFs769), Rodhococcus fascians (DFs843 and DFs912) and Pseudomonas fluorescens (DFs831 and DFs842), selected for Xanthomonas axonopodis pv. phaseoli control, and a combination of some of these bacteria isolates, were evaluated for possible influence on Colletotrichum lindemuthianum transmission from naturally infected and/or infested bean seeds to seedlings. In the first trial, using the paper roll method, seeds were sown in eight replications of 25 seeds that were incubated at 20 ± 2°C. Germination percentage and pathogen incidence were evaluated. In the second trial, pathogen transmission to seedlings was evaluated in sterile substratum and incubated for 10 days. Daily emerged seedlings, pathogen incidence, leaf and root dry mass were determined. An isolate of C. lindemuthianum was used against the biocontroller isolates to evaluate in vitro antibiosis. All of the treatments reduced C. lindemuthianum transmission to seedlings in at least one of the trials. However, isolate DFs912 (Rhodococcus fascians) was shown to be important and presented reductions in transmission that varied from 40 to 67% in all of the experiments. All the treatments promoted leaf and root mass increase on total number or on the number per plant. Results in vitro showed that isolates DFs831, DFs842, DFs843 and DFs912 produced antibiotic compounds.

Biocontrole da queima-das-bainhas e do nematoide-das-galhas e promoção de crescimento de plantas de arroz por rizobactérias

The objective of this work was to evaluate the compatibility between previously selected biocontroller rhizobacteria and the effect of their combination upon sheath blight (Rhizoctonia solani), root-knot nematode (Meloidogyne graminicola) and growth promotion in rice plants. The compatibility between these bacteria was determined by the overlay antibiosis. The effect of isolate combinations was evaluated by seed microbiolization of the rice cultivar El Paso L144 with rhizobacteria suspensions of DFs185 (Pseudomonas synxantha), DFs223 (P. fluorescens), DFs306 (unidentified), DFs416 and DFs418 (Bacillus sp.). These rhizobacteria were used alone or combined in arrangements of two, three and four isolates. The DFs223 isolate was not combined because it is incompatible with the other bacteria. Five rhizobacteria combinations (DFs185/418, DFs306/416, DFs306/418, DFs416/418, DFs185/306/418) and the isolate DFs306 stood out for reducing M. graminicola reproduction and promoting plant growth. Combination DFs185/306 presented the best results for the control of R. solani and for growth promotion. The DFs306/416 combination provided the best results for the control of the two diseases.

Biological 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 (A540=0.5) of each isolate under agitation at 10oC 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.

Potencial fisiológico de sementes de arroz tratadas com rizobactérias ou tiametoxam

Development of new technologies, aiming at increasing productivity in different crops, involves constant research on the effectiveness and application of these techniques in seed treatment. In this study, it was aimed at evaluating physiological potential of rice seeds treated with plant growth-promoting rhizobacteria (PGPR) (strains DFs185, DFs223, DFs306, DFs416), or with two dosages of the insecticide thiamethoxam. The variables assessed were: germination (G); first count of germination (FCG); cold test (CT); length of seedlings aerial parts (LAP), root system (LRS), and total length (TL); emergence speed index (ESI); emergence (E), at 14 days; and dry phytomass (DP). Treatments have had a positive effect on percentage of G, FCG and E. The strain DFs185 has promoted increase in percentage emergence, for five of the six lots assessed. The variables: LAP; LRS; TL; ESI; and DP have undergone low or none influence of treatments; and there has been no toxic effect of rhizobacteria or insecticide thiamethoxam. In the cold test, a negative effect of treatments has been detected. Seed treatment with rhizobacteria, as well as with thiamethoxam, improve quality of low quality rice seeds. The strain DFs185 is promising for treating rice seeds, once it stimulates seed germination and emergence.

Fruit yield and bacterial wilt symptoms on eggplant genotypes grown in soil infested with Ralstonia solanacearum

Fruit yield and bacterial wilt symptoms of eggplant genotypes CNPH006, CNPH171, CNPH658, CNPH778, CNPH783, and CNPH785 were quantified in soil naturally infested with Ralstonia solanacearum (race 1, biovar 3) in Brasilia, Brazil. Except for CNPH778, all genotypes developed at least one typical wilted plant. Besides wilt, other symptoms considered for assessing levels of tolerance/resistance among genotypes were plant death (CNPH006, CNPH171, CNPH658 and CNPH783), leaf chlorosis (CNPH785) and plant dwarfing (all genotypes). The occurrence of bacterial ooze on cut stems was observed in all plants grown in infested beds, including those that did not exhibit wilt symptoms. Plants grown simultaneously in noninfested beds of the same dimensions and soil characteristics allowed the comparison of yield losses due to the pathogen. The genotypes were grouped according to their ability to maintain fruit production in the infested area. CNPH785 was the most tolerant genotype, with no significant loss due to the disease, followed by CNPH783, CNPH778 and CNPH171, with mean losses of 19.3%, 11.4%, and 10.1%, respectively. The genotypes CNPH658 and CNPH006 were the most susceptible, with average losses of 99.53 and 99.32%, respectively.

Bacterial selection for biological control of plant disease: criterion determination and validation

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.

AFLP analysis of Xanthomonas axonopodis and X. arboricola strains used in xanthan production studies reveal high levels of polymorphism

Amplified fragment length polymorphism (AFLP) was used to analyze the genetic diversity of 14 strains of Xanthomonas arboricola pv. pruni and seven strains of X. axonopodis pv. phaseoli, which are used in xanthan production studies. Relationships identified by the AFLP profiles were assessed for xanthan production capacity, geographical location and host plant. Strains were isolated from 10 different geographic regions in South and Southeast States in Brazil. Data were analyzed for genetic similarity using the Dice coefficient and subjected to UPGMA cluster analysis. A total of 128 AFLP fragments were generated from four primer combinations: EcoRI+C/MseI+0, EcoRI+A/MseI+0, EcoRI+G/MseI+T and EcoRI+G/MseI+A. Of these, 96.1% were polymorphic. X. axonopodis pv. phaseoli (SD = 0.27) was shown to be more polymorphic than X. arboricola pv. pruni (SD = 0.58). All 14 pathovar pruni strains were included in a single main group (SD = 0.58), while the pathovar phaseoli strains were divided into three separate groups, with one group containing five strains (SD = 0.38) and two isolated groups (SD = 0.31 and 0.27) composed of only one strain each. Species were distinguished by three and eight specific AFLP markers present in the pathovar phaseoli and the pathovar pruni, respectively. For the unique strain without xanthan production capacity (X. axonopodis pv. phaseoli str. 48), nine specific AFLP bands were found. There was no evidence that geographic area or host plant influenced genetic heterogeneity. Correlations between AFLP patterns and xanthan production capacity were found in some strains, but were not consistent enough to establish a relationship.