Submitted by Helena Bejio (helena.bejio@unioeste.br) on 2017-11-23T17:51:39Z No. of bitstreams: 2 Eloisa Lorenzetti 2017.pdf: 1638897 bytes, checksum: 37d0d636adf093d3df43c201d58eeb5b (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) / Made available in DSpace on 2017-11-23T17:51:39Z (GMT). No. of bitstreams: 2 Eloisa Lorenzetti 2017.pdf: 1638897 bytes, checksum: 37d0d636adf093d3df43c201d58eeb5b (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Previous issue date: 2017-02-10 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPES / Several works using extracts obtained from medicinal plants have demonstrated direct and potential antimicrobial action to induce the accumulation of secondary metabolites, which are very important as plant defense mechanisms against pathogens. The objective of this work was to develop an alternative method using rosemary extract (Rosmarinus officinalis L.) at concentrations 0%, 1%, 2.5% and 5%, to verify the antimicrobial activity against Macrophomina phaseolina, control of charcoal rot in soybean stem and determine the activity of peroxidase, polyphenol oxidase, phenylalanine ammonia-lyase (PAL) and protein content on soybean plants. We performed in vitro assays in order to analyze mycelial growth of the fungus, fungus mass and number of micro-sclerotia, greenhouse tests to evaluate the area under the disease progress curve (AUDPC), and biochemical analyzes to verify a possible induction of resistance in the treated plants. For the determination of the defense enzymes, soybean plants were treated with the extracts and inoculated with M. phaseolina. Samples were collected at 0, 36, 72, 120, 168, 216 and 264 h after treatment. The rosemary extract reduced by 44% and 74% the fungal growth on solid and liquid media, respectively, while for the micro-sclerotia the reduction was of 61%. For the AUDPC in the first and in second assays, there were a reduction of 53% and 56% respectively in the disease. For the samples collected at the base of steam, peroxidase had the highest concentrations of the extract with two peaks of induction. The polyphenol oxidase activity increased from 36 to 120 hours after treatment with extract at 5%. PAL activity was induced only with 5% extract, with increases 83% and 130% higher in times 168 and 216 hours after treatment, respectively. For enzymes activities in root, peroxidase again showed two peaks for increase at 5% concentration, polyphenol oxidase was 426% higher at 216 hours after treatment and PAL showed an increase of 340% at 216 hours after treatment with 5% extract. These results indicate potential of rosemary extract in the control of charcoal rot in soybean and in the induction of plant defense enzymes in soybean. / Diversos trabalhos utilizando extratos obtidos de plantas medicinais têm demonstrado ação antimicrobiana direta e potencial de induzir o acúmulo de metabólitos secundários, sendo estes muito importantes nos principais mecanismos de defesa das plantas aos patógenos. O objetivo deste trabalho foi desenvolver um método alternativo através da utilização do extrato de alecrim (Rosmarinus officinalis L.), nas concentrações 0%, 1%, 2,5% e 5%, para verificar a atividade antimicrobiana contra Macrophomina phaseolina, controlar a podridão cinzenta da haste em soja e determinar as atividades de peroxidase, polifenoloxidase, fenilalanina amônia-liase e teor de proteína de plantas de soja tratadas. Foram realizados ensaios in vitro a fim de analisar o crescimento micelial do fungo, a massa do fungo e o número de microescleródios, ensaios in vivo em casa de vegetação para avaliar a área abaixo da curva de progresso da doença (AACPD), além de análises bioquímicas para verificar uma possível indução de resistência nas plantas tratadas. Para determinação das enzimas de defesa, plantas de soja foram tratadas com os extratos e inoculadas com M. phaseolina, sendo retiradas amostras para análise nos tempos 0, 36, 72, 120, 168, 216 e 264 h após o tratamento. O extrato de alecrim reduziu em até 44% e 74% o crescimento fúngico em meios sólido e líquido, respectivamente, enquanto que para micro-escleródios a redução foi de 61%. Para AACPD, no primeiro e no segundo ensaios houve redução de 53% e 56% respectivamente. Nas amostras retiradas do colo das plantas, para peroxidase, as concentrações mais elevadas do extrato proporcionaram dois picos de indução. Houve constante incremento na atividade de polifenoloxidase desde 36 até 120 h após o tratamento para a concentração 5% do extrato de alecrim. Para FAL apenas a concentração 5% promoveu incremento 83% e 130% maior nos tempos 168 e 216 h após o tratamento, respectivamente. Para as atividades na raiz, a peroxidase novamente apresentou dois picos de incremento para a concentração 5% do extrato, a polifenoloxidase foi 426% maior na concentração 5% às 216 h após o tratamento e a atividade de FAL apresentou incremento de 340% no tempo 216 h após o tratamento com 5% do extrato. Estes resultados indicam potencial do extrato de alecrim no controle de podridão cinzenta da haste em soja e na indução da atividade de enzimas de defesa em colo e raiz de soja.

The production of strawberries can be severely limited by soilborne plant pathogens, insects and weeds. Macrophomina phaseolina is a problematic soilborne fungal pathogen in California strawberry production inciting the disease Macrophomina crown rot. When established, the pathogen can cause extensive plant decline and mortality. Host resistance will be a critical tool for managing this disease and guiding breeding programs in the post methyl bromide era. Evaluation of host resistance in strawberry germplasm to M. phaseolina was evaluated through phenotypic assessments of disease incidence. A total of 90 strawberry cultivars and elite selections were included in a replicated field trial conducted in artificially inoculated soils to assess host resistance. Significant differences in levels of resistance and susceptibility were observed among genotypes tested in this trial. The five most resistant strawberry genotypes from highest to lowest in percent plant mortality were: UC-R, UC-G, UC-V, Manresa, and Osceola. The five most susceptible strawberry genotypes with the highest percent mortality in ranking order from highest to lowest were: UC-J, Ruby June, Festival, UC-Y, and UC-A. Of the genotypes tested in this trial UC-V, Manresa and Osceola could be characterized as highly resistant, but no complete resistance was observed. An additional study was conducted to correlate host symptom expression with the extent of pathogen colonization in different strawberry tissues, and to determine if resistant germplasm can contribute to secondary inoculum production in the field. An established qPCR method was utilized to quantify M. phaseolina colonization of strawberry tissues. There were significant effects for cultivar (P < 0.0001) as well as a significant two-way interaction of cultivar x sample time (P= 0.0083) on the concentration of M. phaseolina DNA detected in strawberry tissues. Expression of the resistant phenotype in strawberry cultivars was associated with limited plant colonization by M. phaseolina. The extent of colonization of a specific cultivar by M. phaseolina was dependent on the sample time after inoculation with the pathogen. In addition, the roots and crowns of a specific strawberry cultivar were equally colonized on a per plant tissue weight basis, but this provides only speculation into the mechanisms conferring host resistance. A third study was conducted to integrate host resistance of strawberry genotypes with the use of organic amendments in effort to mutually enhance the efficacy of each factor for the control of Macrophomina crown rot. Artificially inoculated potting substrate was amended with Brassica juncea mustard seed meal at a rate of 4.94 tons ha-1(MSM), and anaerobic soil disinfestation utilizing rice bran at a rate of 22.24 tons ha-1 (ASD) were compared to a non-amended (UTC) and steam controls. The soil assay indicated that the ASD and steam treatments were able to reduce the CFU g-1 potting substrate of M. phaseolina by 99.7-100%. In addition, there were significant effects of soil treatment on the fresh biomass of weed seedlings recovered from the potting substrate. However, disease severity and host colonization of multiple strawberry cultivars by M. phaseolina was not reduced when grown in the treated potting substrate. The effect of strawberry cultivar on the extent of pathogen colonization was highly significant (P < 0.0001), in which cultivars characterized as resistant from phenotypic screenings possessed lower concentrations of M. phaseolina DNA. The suppression of M. phaseolina in response to organic amendments was limited but this study supports findings from previous experiments that genotype specific host resistance minimizes host colonization and reduces the production of secondary inoculum.

Submitted by Johnny Rodrigues (johnnyrodrigues@ufcg.edu.br) on 2018-05-03T18:31:57Z No. of bitstreams: 1 ANDRESSA LACERDA NÓBREGA UGULINO - DISSERTAÇÃO PPGSA ACADÊMICO 2017..pdf: 676544 bytes, checksum: 0dd2e41c1011a60929ca0702449f6287 (MD5) / Made available in DSpace on 2018-05-03T18:31:57Z (GMT). No. of bitstreams: 1 ANDRESSA LACERDA NÓBREGA UGULINO - DISSERTAÇÃO PPGSA ACADÊMICO 2017..pdf: 676544 bytes, checksum: 0dd2e41c1011a60929ca0702449f6287 (MD5) Previous issue date: 2017 / O fungo Macrophomina phaseolina (Tassi.). Goid é o agente etiológico de uma das principais doenças na cultura do feijão-caupi, vulgarmente conhecida como podridão cinzenta do caule. Em decorrência da falta de produtos fitossanitários registrados para o controle desta doença um aumento na busca por métodos de controle alternativo é cada vez mais comum. O objetivo do presente trabalho foi avaliar o efeito de óleos essenciais sobre o crescimento micelial do fungo M. phaseolina. Os óleos testados foram de hortelã (Mentha sp.), eucalipto (Eucalyptus spp.), copaíba ( Copaifera sp.) e alecrim-da-chapada (Lippia gracilis), nas concentrações 0,4; 0,6; 0,8 e 1,0%. Os óleos foram incorporados ao meio de cultura do tipo BDA e posteriormente vertidos em placas de Petri de 90x15mm, após a solidificação do meio, discos de 8mm contendo o micélio do patógeno foram repicados para as placas com os respectivos tratamentos. O experimento foi conduzido no delineamento experimental inteiramente casualizado (DIC) em arranjo fatorial 4 x 4 + 1 +1, sendo dezesseis tratamentos mais uma testemunha negativa e uma testemunha positiva que constou da aplicação suplementar no meio (BDA) do fungicida Sportak 450 EC (Procloraz), e cinco repetições, totalizando 90 parcelas experimentais. Posteriormente todas as placas foram transferidas para incubadora do tipo B.O.D. (biochemical oxygen demand) a 27 ± 2 ºC. Foram realizadas medições diárias do diâmetro da colônia em dois sentidos perpendiculares até que tomasse toda a superfície do meio de cultura de uma das placas. Os dados obtidos foram interpretados por meio de análise de variância não paramétrica e as médias foram comparadas pelo teste de Kruskal-wallis, a 5% de probabilidade de erro e Scott Knott a 5% de probabilidade nos casos em que existiram diferenças significativas e normalidade dos dados. Os óleos essenciais de hortelã e alecrimda-chapada apresentaram melhores resultados quanto a inibição do crescimento micelial, enquanto que os óleos de eucalipto e copaíba apesar de potencialmente promissores, apresentaram inibição intermediária do crescimento micelial do fungo. / Macrophomina phaseolina (Tassi.). Goid is the etiological agent of one of the major diseases in cowpea bean crop, commonly known as gray rot stem. Due to the lack of registered phytosanitary products to control this disease, the search for alternative control methods is increasingly common. The objective of this study was to evaluate the effect of essential oils on the mycelial growth of M. phaseolina. The oils tested were Mentha sp.,Eucalyptus spp., Copaifera sp., and Lippia gracilis at concentrations of 0.4; 0.6; 0.8 and 1.0%. The work was conducted in Phytopathology Laboratory of the Federal University of Campina Grande. Daily measurements of the colony diameter were performed in two perpendicular directions until it filled the entire surface of the culture medium of one of the plates. The experiment was conducted in a completely randomized experimental design in a factorial arrangement 4 x 4 + 1+1, with sixteen treatments plus one negative control and one positive control, which consisted of the supplemental application in the medium (BDA) of the fungicide Sportak 450 EC (Procloraz), and five replications, totaling 90 experimental plots. The data were interpreted through non-parametric analysis of variance and the means were compared by the Kruskal-wallis test, with a 5% probability of error and Scott Knott at 5% probability in cases where there were significant differences and data normality. The Mentha sp. essential oil and Lippia gracilis showed better results in inhibiting mycelial growth, while the Eucalyptus essential oil and Copaifera, although potentially promising, showed intermediate inhibition of fungal mycelial growth.