Submitted by Mario BC (mario@bc.ufrpe.br) on 2016-11-28T12:59:54Z No. of bitstreams: 1 Dyana de Albuquerque Tenorio.pdf: 1359840 bytes, checksum: 2d9414b172e6c51ef868267286f6ef2b (MD5) / Made available in DSpace on 2016-11-28T12:59:54Z (GMT). No. of bitstreams: 1 Dyana de Albuquerque Tenorio.pdf: 1359840 bytes, checksum: 2d9414b172e6c51ef868267286f6ef2b (MD5) Previous issue date: 2015-02-25 / The damping-off and stem rot caused by the soil pathogen Rhizoctonia solani, is considered one of the most important diseases of common bean (Phaseolus vulgaris) and cowpea (Vigna unguiculata), being one of the factors that contribute to the low productivity of these crops. This study aimed to characterize isolates of the causal agent of damping-off and stem rotof beans in the rural area of Pernambuco State, using microsatellite markers and phylogenetic analysis; and evaluate the use of yeasts in the control of the causal agent of damping-off and stem rot of beans. In bean growing areas of the southern region of Pernambuco, there were collections of plants with symptoms of damping-off and stem rot. Thirty-four fungal isolates were identified as Rhizoctonia. In the phylogenetic analysis, the sequences of ITS rDNA region of isolates were similar to at least three anastomosis groups (AG4 HG-I, AG3-PT and AG4 HG-III). In microsatellite analysis, five sets of primers (TC_AG3_1, TC01, TC06, TC11 and TC_AG3_09) amplified a total of 13 bands, showing genetic variation. Seventy R. solani were evaluated for aggressiveness, being the isolate CMM-3643 selected to the biological control trials. Seventy yeast isolates were obtained from apparently asymptomatic bean plants. From these, were chosen only isolates where the disease severity index was ≤ 25%. Experimental tests showed that the three yeast isolates (C6A, FVF10 (R1), FVC10) were selected as promising biocontrol agents of damping-off and stem rot of cowpea (cv. IPA-207), being efficient in controlling the disease caused by isolated CMM -3643. Increases in the levels of POX and CAT enzymes by treating the seeds with FVF10 yeast (R1), as well as, the absence of antibiosis mechanisms suggests that the biocontrol mechanism in question is the induction of disease resistance. In view of the results it is concluded that the AG4 HG-I is the primary anastomosis group causing damping-off and stem rot of beans in the São João municipality, Pernambuco State. In addition, this is the first report of damping-off and stem rot symptoms reduction in cowpea seedlings due to the antagonistic action of yeasts against R. solani. / A rizoctoniose, causada pelo fungo de solo Rhizoctonia solani, é considerada uma das doenças mais importantes do feijoeiro-comum (Phaseolus vulgaris) e do feijoeiro-caupi (Vigna unguiculata), sendo um dos fatores responsáveis pela baixa produtividade da cultura. O presente trabalho teve como objetivos: caracterizar isolados do agente causal da rizoctoniose do feijoeiro na região do agreste de Pernambuco, utilizando marcadores microssatélites e análise filogenética; e avaliar o uso de leveduras no controle do agente causal da rizoctoniose do feijoeiro. Em áreas de cultivo do feijoeiro da região do agreste meridional de Pernambuco, realizaram-se coletas de plantas com sintomas da rizoctoniose. Sendo obtidos 34 isolados fúngicos identificados como Rhizoctonia. Na análise filogenética, as sequências gênicas da região ITS do rDNA destes isolados foram similares a pelo menos três grupos de anastomose (AG4 HG-I, AG3-PT e AG4 HG-III). Na análise de microssatélites, cinco conjuntos de iniciadores (TC_AG3_1, TC01, TC06, TC_AG3_09 e TC11) amplificaram um total de 13 bandas, observando-se variação genética. Setenta isolados de R. solani foram avaliados quanto a agressividade, sendo selecionado o isolado CMM-3643 para o ensaio de controle biológico. Foram obtidos 70 isolados de leveduras de plantas de feijoeiro aparentemente assintomáticas, destes foram escolhidos os que proporcionaram índice de severidade da doença ≤ 25%. Ensaios experimentais demonstraram que os três isolados de leveduras (C6A, FVF10 (R1), FVC10) selecionados foram promissores como agentes de biocontrole da rizoctoniose do feijoeiro-caupi (cv. IPA-207), apresentando eficiência no controle da doença causada pelo isolado CMM-3643. Aumentos nos níveis das enzimas POX e CAT pelo tratamento das sementes com a levedura FVF10 (R1), assim como a ausência de mecanismos de antibiose, sugere que o mecanismo de biocontrole em questão é a indução de resistência da planta. Com base nas avaliações realizadas concluiu-se que o AG4 HG-I é o principal grupo de anastomose da rizoctoniose do feijoeiro no município de São João. Além disso, este é o primeiro relato da redução dos sintomas da rizoctoniose em plântulas de feijoeiro-caupi pela ação antagônica de leveduras sobre R. solani.

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Orientador: Paulo Cezar Ceresini / Resumo: Rhizoctonia solani AG-1 IA é um patógeno habitante do solo que causa doenças em pastagens e plantas cultivadas, em vários estados brasileiros. Para penetrar e colonizar os tecidos do hospedeiro, o microrganismo utiliza enzimas que podem ser de interesse biotecnológico e industrial. O objetivo do presente trabalho foi produzir e caracterizar a α-amilase secretada por R. solani AG-1 IA utilizando produtos e resíduos agroindustriais como substratos para fermentação. Para todos os ensaios foi adotado o processo de cultivo em estado sólio (CES) à temperatura de 25 °C e umidade inicial de 60%. Inicialmente, nove isolados de R. solani provenientes de diferentes hospedeiros (arroz, braquiária e soja) foram comparados quanto à produção amilolítica. As maiores atividades enzimáticas foram proporcionadas pelos isolados TO_022 e MT_SO85, procedentes da cultura da soja. Na etapa seguinte, foi avaliado o efeito do substrato sobre a produção enzimática. Farelo de trigo (12,98 U mL-1; TO_022), farelo de soja (3,97 U mL-1; MT_SO85) e braquiária lavada (4,95 U mL-1; TO_022) induziram eficientemente a produção amilolítica. O perfil de produção enzimática no farelo de trigo, melhor substrato avaliado no cultivo, indicou o tempo de 216 h como o mais apropriado para a obtenção de α-amilases pelo isolado TO_022 (22,14 U mL-1), e 240 h para o isolado MT_SO85 (22,84 U mL-1). Para caracterização físico-química da enzima foi utilizado o extrato enzimático bruto do isolado TO_022. A α-amilase de R. sola... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: Rhizoctonia solani AG-1 IA is a soil-borne pathogen that causes diseases in pastures and cultivated plants in several Brazilian states. To penetrate and colonize host tissues, the microorganism uses enzymes that may be of biotechnological and industrial interest. The aim of the present work was to produce and characterize the α-amylase secreted by R. solani AG-1 IA using agroindustrial products and residues as substrates for fermentation. For all the tests, the solid state cultivation process (SSC) at 25 °C and initial humidity of 60% was adopted. Initially, nine isolates of R. solani from different hosts (rice, Brachiaria grass and soybean) were compared for amylolytic production. The major enzymatic activities were provided by the isolates TO_022 and MT_SO85, from the soybean crop. In the next step, the effect of the substrate on the enzymatic production was evaluated. Wheat bran (12.98 U mL-1; TO_022), soybean meal (3.97 U mL-1, MT_SO85) and washed ruzigrass (4.95 U mL-1; TO_022) efficiently induced amylolytic production. The enzymatic production profile in wheat bran, the best substrate evaluated in the culture, indicated the time of 216 h as the most appropriate to obtain α-amylases by the TO_022 isolate (22.14 U mL-1), and 240 h for the isolated MT_SO85 (22.84 U mL-1). For the physicochemical characterization of the enzyme, the crude enzymatic extract of the TO_022 isolate was used. The α-amylase of R. solani TO_022 exhibited maximum activity at 55 °C and pH 5.5, and ma... (Complete abstract click electronic access below) / Mestre

Currently, 74% of cotton acres in the United States are planted with glyphosatetolerant varieties. The average glyphosate-tolerant cotton crop is treated with glyphosate 2.1 times each year in addition to other herbicides, insecticides, and fungicides. The primary objectives of this research were to: 1) describe the influence of glyphosate and pesticides commonly applied at or near the time of cotton planting on soil microbial activity and biomass; 2) study the effect of glyphosate on fluometuron degradation; 3) evaluate the response of Rhizoctonia solani to glyphosate and fluometuron; 4) study changes in glyphosate metabolism that occur as a result of repeated glyphosate applications; and 5) define shifts in the soil microbial community. Additionally, methods for accelerated solvent extraction (ASE) of fluometuron from soils were developed. In one experiment, the addition of glyphosate reduced C-mineralization in soils treated with fluometuron, aldicarb, or mefenoxam + PCNB formulations. However, in a second experiment, C-mineralization increased when glyphosate was applied with fluometuron relative to fluometuron applied alone. Accelerated solvent extraction was used in experiments which demonstrated that application of glyphosate with fluometuron increased the rate of fluometuron degradation in soil relative to fluometuron alone. When glyphosate was added to minimal medium, degradation of fluometuron by R. solani was reduced and less fungal biomass was produced. The total amount of 14C-glyphosate mineralized was reduced when glyphosate was applied 5 times relative to 1, 2, 3, or 4 times. Incorporation of 14Cglyphosate residues into soil microbial biomass was greater following five glyphosate applications than one application 3 and 7 days after application (DAA). Soil fatty acid methyl ester (FAME) profiles were altered by five glyphosate applications relative to one application. Additionally, FAMEs common to gram-negative bacteria were present in higher concentrations following five applications relative to 1, 2, 3, or 4 applications both 7 and 14 DAA. These studies indicated that: 1) glyphosate altered the soil microbial response to other pesticides; 2) fluometuron-degrading microorganisms in soil responded differently to glyphosate; 3) changes in the dissipation or distribution of glyphosate following repeated glyphosate applications were associated with changes in the structural diversity of the soil microbial community.

The increasing concerns about chemical pesticides that are environmentally hazardous and the continuous development of resistance by palhogens to chemical pesticides have led to this study. Many studies have shown that some Gram-negative bacteria, such as Pseudomonas flouresens, control plant diseases and promote plant growth. In this study Gram positive bacteria, Bacillus sp., were chosen because of their ability to produce endospores. Endospores can be used in stable, dry formulations. The advantage of using endospores is their ability to survive harsh conditions such as droughts and high temperatures, which give a long shelf life to the biological control agent. Bacillus isolates were recovered from the rhizosphere of 12 different crops, and were subsequently screened in vitro for their antimicrobial activity. Of 130 isolates, 87 exhibited antimicrobial activity against the test organisms: Rhizoctonia solani, Pythium sp., Phytophthora cinnamoni, Fusarium sp., and single representatives of Gram negative and Gram positive bacteria, namely, Erwinia carotovora and Staphylococcus aureus respectively. The Bacillus isolates B77, B81 and B69 inhibited all the test organisms investigated, which suggests that they produced broad spectrum antimicrobial compounds or more than one antimicrobial compound. Of the isolates that showed antimicrobial activity, 78 of them did not inhibit Trichoderma harzianum K D, which is a registered biological control agent; indicating their potential for combined application. Selected Bacillus isolates were tested for the biological control of R. solani under greenhouse conditions in wheat, cabbage, tomato, maize, and cucumber seedlings. Bacillus isolates were applied as seed treatments, and the inoculated seeds were planted in R. solani infested speedling trays. Shoot dry weight measurement of seedlings indicated that 12 out of 19 Bacillus isolates showed significantly different shoot dry weight in wheat whereas all the isolates tested in tomato and cucumber gave significantly different shoot dry weight. No significantly different shoot dry weight was obtained for maize or cabbage. Seed emergence findings indicated that none of the Bacillus isolates gave significantly different emergence percentage on wheat, cabbage, tomato, and maize but all of them showed significantly different emergence percentage on cucumber. The results indicate that both the pathogen and the biological control agents exhibited varying levels of specificity on each crop tested. The biological control potential of the best Bacillus isolates was tested on bean and maize crops in the field. Green bean and maize seeds were coated with the selected Bacillus isolates and then sown under field conditions. For each isolate, four replicate treatment plots were established, with and without a R. solani inoculum. Percentage emergence, plant survival levels to harvesting and yield of maize cobs and green beans pods were measured. For all parameters measured the positive and negative controls were not significantly different thereby rendering the results for the entire field study inconclusive. However, Bacillus isolates B77, BII, R5 and R7 improved green bean pod yield and Bacillus Isolate B8I increased maize yield, indicating their potentials as plant growth promoting rhizobacteria (PGPR). / Thesis (M.Sc.)-University of Natal, Pietermaritzburg, 2003.

Made available in DSpace on 2014-06-11T19:28:28Z (GMT). No. of bitstreams: 0 Previous issue date: 2009-02-17Bitstream added on 2014-06-13T20:18:13Z : No. of bitstreams: 1 gomes_dp_me_jabo.pdf: 375333 bytes, checksum: d2044e94be55c12e13ab4e37ad5eb7ea (MD5) / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / O objetivo da pesquisa foi determinar a influência de fatores abióticos e bióticos sobre a ocorrência de “damping-off” de pré e de pós – emergência, além de outros danos nas sementes e plântulas, quando da semeadura de milho e feijão. Foram escolhidos dois lotes de milho e feijão, safra 2007/2008, provenientes das cultivares D 766 e Carioca, respectivamente, de acordo com sua distinção em alto e baixo vigor, com baixa incidência fúngica e ausência de Rhizoctonia solani. Sementes desinfestadas foram infectadas com o fungo em meio de cultura BDA com restrição hídrica (acrescido de manitol a -1,0 MPa). Nos tratamentos testemunha foi utilizado o mesmo meio de cultura, porém, sem a presença do fungo. Para cada tratamento as sementes foram sobrepostas nos meios de cultura por 36 horas para o milho e por 20 horas para o feijão. Para a infestação direta do fungo em solo, foram utilizadas sementes de sorgo esterilizadas. O experimento foi conduzido em duas épocas distintas (caracterizadas como fria e quente). Para as duas épocas, as parcelas experimentais foram constituídas de vasos plásticos (capacidade de 4,5 L) contendo terra esterilizada dispostos segundo a casualização dos tratamentos em casa de vegetação. Os tratamentos foram constituídos a partir de três disponibilidades hídricas no solo contido em vaso (30 %, 50 % e 70 % de retenção de água), dois níveis de vigor da semente (alto e baixo vigor) com e sem a inoculação de Rhizoctonia solani, através de inoculação na própria semente do hospedeiro e da infestação no solo através de sementes de sorgo esterilizadas. Foram feitas quatro repetições, cada uma contendo dois vasos, totalizando oito vasos para cada tratamento. / The objective of the research was to determine the influence of biotic and abiotic factors on the occurrence of damping-off of pre and post - emergence, and others problems to the seeds and seedlings, when the sowing of corn and bean. They chose two plots of corn and bean, growing season 2007/2008, from the cultivars Carioca and D 766 respectively, according to his distinction in high and low vigour, low fungi incidence and absence of Rhizoctonia solani. Disinfested seeds were infected with the fungus in culture medium without BDA and water restriction (mannitol, -1.0 MPa). In control treatments was the same growing medium, but without the presence of the fungus. For each treatment the seeds were superimposed in culture media for 36 hours for the corn and bean for 20 hours. For the direct addition of the fungus in soil, we used the technique of sterile sorghum seeds. The experiment was conducted in two different seasons (hot and cold). For two growing seasons the plots were established in plastic pots (capacity 4.5 L) containing sterilized soil prepared according to the randomization of treatments in a greenhouse. The treatments were made from three water availability in the soil contained in vase (30 %, 50 % and 70 % retention of water), two levels of vigour of the seed (high and low vigour) with and without inoculation of Rhizoctonia solani by of inoculation in the seed of the host and soil addition with sterile sorghum seeds. Were four replicates, each containing two vessels, totaling eight pots for each treatment. Twenty days after inoculation, for the cold season, and fifteen days for the hot season was to assess the severity, damping-off of pre and post-emergence, seedling length, dry matter weight and percentage of seedlings healthy.