15 research outputs found
Caracterização morfológica, bioquímica e proteomica da embriogênese zigótica e somática de goiabeira serrana (Acca sellowiana (O. Berg.) Burret)
Tese (doutorado) - Universidade Federal de Santa Catarina. Centro de Ciências Agrárias. Programa de Pós-graduação em Recursos Genéticos VegetaisNesta seção, estudaram-se as proteínas expressas em embriões somáticos de A. sellowiana em diferentes estádios de desenvolvimento. O objetivo foi detectar e identificar proteínas diferencialmente expressas durante os diferentes estádios de desenvolvimento do embrião somático. Usando a alta resolução do gel bidimensional de poliacrilamida (2-DE) observaram-se diferenças entre as proteínas expressas nos estádios iniciais e similaridade entre as proteínas expressas nos últimos estádios. Foram detectadas 29, 32, 51, 61 e 57 proteínas para o estádio globular, cordiforme, torpedo, pré-cotiledonar e cotiledonar, respectivamente. Das 230 proteínas visualizadas por 2-DE, 62 foram identificadas por MALDI TOF/MS. Quatro proteínas foram expressas unicamente no estádio globular, uma no estádio cordiforme, duas no estádio torpedo e quatro no estádio pré-cotiledonar. As proteínas identificadas nos diferentes estádios da embriogênese somática foram agrupadas nas seguintes categorias: metabolismo de carboidratos, biossíntese de purinas, divisão celular, metabolismo secundário, reserva, chaperonas, formação e transporte celular. Os estudos de proteoma com esta espécie são inéditos e permitem avançar no conhecimento do metabolismo embrionário desta espécie, bem como possibilitam uma adequação dos protocolos de embriogênese somática visando sua propagação massal e conservação
Padrões de acúmulo de proteínas e carboidratos durante a embriogênese somática de Acca sellowiana
The aim of this work was to quantify the protein, starch and total sugars levels during histodifferentiation and development of somatic embryos of Acca sellowiana Berg. For histological observations, the samples were dehydrated in a battery of ethanol, embedded in historesin and stained with toluidine blue (morphology), coomassie blue (protein bodies) and periodic acid-Schiff (starch). Proteins were extracted using a buffer solution, precipitated using ethanol and quantified using the Bradford reagent. Total sugars were extracted using a methanol-chloroform-water (12:5:3) solution and quantified by a reaction with anthrone at 0.2%. Starch was extracted using a 30% perchloric acid solution and quantified by a reaction with anthrone at 0.2%. During the somatic embryogenesis' in vitro morphogenesis and differentiation processes, the total protein levels decreased and the soluble sugars levels increased during the first 30 days in culture and remained stable until the 120th day. On the other hand, total protein levels increased according to the progression in the developmental stages of the somatic embryos. The levels of total sugars and starch increased in the heart and cotyledonary stages, and decreased in the torpedo and pre-cotyledonary stages. These compounds play a central role in the development of somatic embryos of Acca sellowiana. © 2009 Embrapa Informação Tecnológica.O objetivo deste trabalho foi quantificar os teores de proteína, amido e açúcares totais durante a histodiferenciação e desenvolvimento dos embriões somáticos em Acca sellowiana Berg. Para as observações histológicas, as amostras foram desidratadas em uma bateria de etanol, emblocadas em historesina e coradas com azul de toluidina (morfologia), azul de coomassie (corpos proteicos) e reativo ácido periódico de Schiff (amido). As proteínas foram extraídas usando uma solução tampão, precipitadas usando etanol e quantificadas por meio do reativo de Bradford. Os açúcares totais foram extraídos usando uma solução metanol-clorofórmioágua (12:5:3) e quantificados pela reação com antrona a 0,2%. O amido foi extraído usando uma solução de ácido perclórico a 30% e quantificado pela reação com antrona a 0,2%. Durante a diferenciação e morfogênese in vitro da embriogênese somática, os teores de proteínas totais decresceram e os açúcares solúveis aumentaram durante os 30 primeiros dias em cultura e permaneceram constantes até os 120 dias. Por outro lado, os teores das proteínas totais apresentaram incremento de acordo com a progressão nos estádios de desenvolvimento dos embriões somáticos. Os teores de açúcares totais e de amido aumentaram nos estádios cordiforme e cotiledonar e diminuíram nos estádios torpedo e pré-cotiledonar. Esses compostos exercem papel central no desenvolvimento de embriões somáticos de Acca sellowiana.Fil: Cangahuala-Inocente, Gabriela Claudia. Universidade Federal de Santa Catarina; BrasilFil: Steiner, Neusa. Universidade Federal de Santa Catarina; BrasilFil: Maldonado, Sara Beatriz. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Biodiversidad y Biología Experimental y Aplicada. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Biodiversidad y Biología Experimental y Aplicada; ArgentinaFil: Guerra, Miguel Pedro. Universidade Federal de Santa Catarina; Brasi
Padrões de acúmulo de proteínas e carboidratos durante a embriogênese somática de Acca sellowiana
The aim of this work was to quantify the protein, starch and total sugars levels during histodifferentiation and development of somatic embryos of Acca sellowiana Berg. For histological observations, the samples were dehydrated in a battery of ethanol, embedded in historesin and stained with toluidine blue (morphology), coomassie blue (protein bodies) and periodic acid–Schiff (starch). Proteins were extracted using a buffer solution, precipitated using ethanol and quantified using the Bradford reagent. Total sugars were extracted using a methanol-chloroform-water (12:5:3) solution and quantified by a reaction with anthrone at 0.2%. Starch was extracted using a 30% perchloric acid solution and quantified by a reaction with anthrone at 0.2%. During the somatic embryogenesis’ in vitro morphogenesis and differentiation processes, the total protein levels decreased and the soluble sugars levels increased during the first 30 days in culture and remained stable until the 120th day. On the other hand, total protein levels increased according to the progression in the developmental stages of the somatic embryos. The levels of total sugars and starch increased in the heart and cotyledonary stages, and decreased in the torpedo and pre-cotyledonary stages. These compounds play a central role in the development of somatic embryos of Acca sellowiana.O objetivo deste trabalho foi quantificar os teores de proteína, amido e açúcares totais durante a histodiferenciação e desenvolvimento dos embriões somáticos em Acca sellowiana Berg. Para as observações histológicas, as amostras foram desidratadas em uma bateria de etanol, emblocadas em historesina e coradas com azul de toluidina (morfologia), azul de coomassie (corpos proteicos) e reativo ácido periódico de Schiff (amido). As proteínas foram extraídas usando uma solução tampão, precipitadas usando etanol e quantificadas por meio do reativo de Bradford. Os açúcares totais foram extraídos usando uma solução metanol-clorofórmio-água (12:5:3) e quantificados pela reação com antrona a 0,2%. O amido foi extraído usando uma solução de ácido perclórico a 30% e quantificado pela reação com antrona a 0,2%. Durante a diferenciação e morfogênese in vitro da embriogênese somática, os teores de proteínas totais decresceram e os açúcares solúveis aumentaram durante os 30 primeiros dias em cultura e permaneceram constantes até os 120 dias. Por outro lado, os teores das proteínas totais apresentaram incremento de acordo com a progressão nos estádios de desenvolvimento dos embriões somáticos. Os teores de açúcares totais e de amido aumentaram nos estádios cordiforme e cotiledonar e diminuíram nos estádios torpedo e pré‑cotiledonar. Esses compostos exercem papel central no desenvolvimento de embriões somáticos de Acca sellowiana
Root Proteomic Analysis of Grapevine Rootstocks Inoculated with Rhizophagus irregularis and Fusarium oxysporum f. sp. herbemontis
Root Proteomic Analysis of Grapevine Rootstocks Inoculated with Rhizophagus irregularis and Fusarium oxysporum f. sp. herbemontis
Grapevine decline and death caused by the pathogenic fungus Fusarium oxysporum f. sp. herbemontis is among the main phytosanitary problem for viticulture in southern Brazil. The eradication of infected plants is presently the most common procedure for disease control in vineyards. Inoculation with arbuscular mycorrhizal fungi is an option to reduce or neutralize the negative impacts of soil pathogenic microorganisms, but the mechanisms of plant response involved in this process are not yet completely elucidated. In order to better understand these mechanisms, an experiment was carried out to identify proteins related to plant defence induced by the mycorrhizal fungus after infection with the pathogenic fungus. We used the grapevine rootstocks SO4 and R110 (susceptible and resistant to the pathogenic fungus, respectively) inoculated or not inoculated with the mycorrhizal fungus Rhizophagus irregularis, and inoculated or not inoculated with Fusarium oxysporum f. sp. herbemontis. Growth of the rootstocks’ shoot and root and presence of pathogenic symptoms were evaluated. The protein profiles of roots were characterized by two-dimensional electrophoresis and proteins were identified using mass spectrometry. The grapevine rootstocks inoculated with R. irregularis had higher biomass production and lower level of pathogenic symptoms. The R110 rootstock differentially accumulated 73 proteins, while SO4 accumulated 59 proteins. Nine plant-defence proteins were expressed by SO4 rootstock, and six were expressed by R110 rootstock plants. The results confirm the effect of mycorrhizal fungi in plant growth promotion and their potential for biological control against soil pathogenic fungus. Protein expression is dependent on rootstock characteristics and on the combination of plant material with the fungi
Root Proteomic Analysis of Grapevine Rootstocks Inoculated with Rhizophagus irregularis and Fusarium oxysporum f. sp. herbemontis
ABSTRACT Grapevine decline and death caused by the pathogenic fungus Fusarium oxysporum f. sp. herbemontis is among the main phytosanitary problem for viticulture in southern Brazil. The eradication of infected plants is presently the most common procedure for disease control in vineyards. Inoculation with arbuscular mycorrhizal fungi is an option to reduce or neutralize the negative impacts of soil pathogenic microorganisms, but the mechanisms of plant response involved in this process are not yet completely elucidated. In order to better understand these mechanisms, an experiment was carried out to identify proteins related to plant defence induced by the mycorrhizal fungus after infection with the pathogenic fungus. We used the grapevine rootstocks SO4 and R110 (susceptible and resistant to the pathogenic fungus, respectively) inoculated or not inoculated with the mycorrhizal fungus Rhizophagus irregularis, and inoculated or not inoculated with Fusarium oxysporum f. sp. herbemontis. Growth of the rootstocks’ shoot and root and presence of pathogenic symptoms were evaluated. The protein profiles of roots were characterized by two-dimensional electrophoresis and proteins were identified using mass spectrometry. The grapevine rootstocks inoculated with R. irregularis had higher biomass production and lower level of pathogenic symptoms. The R110 rootstock differentially accumulated 73 proteins, while SO4 accumulated 59 proteins. Nine plant-defence proteins were expressed by SO4 rootstock, and six were expressed by R110 rootstock plants. The results confirm the effect of mycorrhizal fungi in plant growth promotion and their potential for biological control against soil pathogenic fungus. Protein expression is dependent on rootstock characteristics and on the combination of plant material with the fungi