La formation de troubles physico-chimiques pendant le stockage de boissons clarifiées préoccupe la filière cidricole. Ces troubles sont dus à des interactions entre différents constituants de la boisson, générant des agrégats visibles. Ce travail présente un double objectif : étudier la composition des troubles afin de déterminer les familles chimiques impliquées, puis étudier les mécanismes des interactions responsables de l’apparition de ces troubles. Pour cela, la composition des troubles a été analysée dans trois boissons cidricoles (cidre, jus de pomme et pommeau). Les résultats ont montré l’implication des composés phénoliques et ont conduit à l’hypothèse selon laquelle l’oxydation de ces composés jouerait un rôle prépondérant dans leur agrégation. Des protéines ont également été dosées en grandes concentrations dans des troubles de jus de pomme, suggérant leur implication dans leur formation.Ces hypothèses ont été vérifiées par deux approches en solutions modèles : en modèle pommeau et en modèle jus de pomme. Ces travaux ont mis en évidence des troubles de différentes natures en fonction de la boisson étudiée. D’une part, les troubles des cidres et des pommeaux s’expliqueraient essentiellement par l’auto-agrégation des procyanidines oligomères consécutive à leur oxydation. Les marqueurs moléculaires impliqués dans la formation de trouble réversible ont pu être identifiés. D’autre part, les troubles de certains jus de pomme, relativement pauvres en composés phénoliques et riches en protéines, seraient provoqués par la dénaturation de protéines de défense des plantes / Physico-chemical haze appearance during storage of clarified apple-based beverages is a concern for producers. These hazes are caused by interactions between several constituents of the beverage that lead to the formation of visible aggregates. This work had two main goals: analyze the composition of hazes in order to determine which families of compounds are responsible for their formation, and understand which mechanisms are involved. First, the composition of the haze gathered from three apple-based beverages (cider, apple juice and pommeau) was analyzed. The results revealed the implication of phenolic compounds and led to the hypothesis that their oxidation was probably one of the main factors responsible for haze formation. Proteins were found in quite large quantities in some apple juice hazes, which suggests their involvement in haze formation in this beverage.These two hypotheses have been verified using two model approaches: in a model pommeau and in a model apple juice. This work evidenced that different kinds of hazes exist in apple-based beverages. On the one hand, haze in pommeaux and ciders is mainly explained by procyanidin oligomers self-aggregation induced by their oxidation, with possible interactions with other beverage constituents. On the other hand, haze in some apple juices, which probably contain low polyphenol and high protein levels, is triggered by “Pathogenesis-Related Proteins” denaturation that lead to their self-aggregation, in interaction with oligomeric procyanidins.

Made available in DSpace on 2017-07-10T17:37:26Z (GMT). No. of bitstreams: 1 Sandra Luisa Toillier.pdf: 545173 bytes, checksum: 7cdf4b17689b7059b1098820885d4e6c (MD5) Previous issue date: 2008-08-01 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / Bean (Phaseolus vulgaris) is among one of the main Brazilian crops and can be affected by several diseases, such as common bacterial blight caused by Xanthomonas axonopodis pv. phaseoli. The study of this interaction can contribute to development of alternative methods to control this disease. The aim of this work was to verify the antimicrobial and resistance induction activities of Pycnoporus sanguineus extracts for the control of this bean disease. The in vitro assays was conducted at the Laboratory of Plant of the State University of West of Paraná (UNIOESTE) - Campus of Marechal Cândido Rondon - PR and were used aqueous extracts from basidiocarp, mycelium and culture filtrate of P. sanguineus in concentrations of 1, 5, 10, 15 and 20%, with water, acibenzolar-S-methyl (ASM - 125 mg a.i. L-1) and antibiotic (oxytetracycline 22.5 mg L-1 + streptomycin 225 mg L-1) as control treatments. For in vivo assays were evaluated disease severity and the activities of peroxidase, polyphenol oxidase, β-1,3 glucanase and phenylalanine ammonia-lyase, using extracts of mycelium, basidiocarp and culture filtrate of P. sanguineus at 5 and 10%, these tests being conducted in greenhouses in the area belonging to the Complex Biological Control and Protected Cultivation Mario Cesar Lopes of UNIOESTE, Campus of Marechal Cândido Rondon. In vitro was verified antibacterial activity only for culture filtrate in concentrations up 15% and for all concentrations of basidiocarp extract. In vivo the results indicated the potential of basidiocarps extracts from P. sanguineus for the control of Xanthomonas axonopodis pv. phaseoli in bean, which can occur either by direct antimicrobial activity and resistance induction involving the activation of enzymes studied / A cultura do feijão (Phaseolus vulgaris) está entre uma das principais da agricultura nacional e pode ser afetada por várias doenças, como o crestamento bacteriano comum causado por Xanthomonas axonopodis pv. phaseoli. O entendimento desse patossistema pode contribuir para que sejam desenvolvidos métodos alternativos para o controle desta doença. O objetivo deste trabalho foi verificar as atividades antibacteriana e indutora de resistência de extratos de Pycnoporus sanguineus para controle desta doença em feijoeiro. O estudo in vitro foi realizado no Laboratório de Fitopatologia da Universidade Estadual do Oeste do Paraná (UNIOESTE) Campus de Marechal Cândido Rondon PR e foram utilizados extratos aquosos de basidiocarpo, micélio e filtrado de cultura de P. sanguineus nas concentrações de 1, 5, 10, 15 e 20%, além das testemunhas, água, acibenzolar-S-metil (ASM - 125 mg i.a. L-1) e antibiótico (22,5 mg L-1 de oxitetraciclina + 225 mg L-1 de estreptomicina). Para o teste in vivo, foi realizada a avaliação de severidade e atividade de peroxidase, polifenoloxidase, β-1,3 glucanase e fenilalanina amônia-liase, com o uso de extrato de micélio, basidiocarpo e filtrado de cultura de P. sanguineus a 5 e 10%, sendo estes ensaios conduzidos em estufa na área pertencente ao Complexo de Controle Biológico e Cultivo Protegido Mário César Lopes da UNIOESTE, Campus de Marechal Cândido Rondon. In vitro verificou-se atividade antibacteriana apenas para o filtrado de cultura em concentrações acima de 15% e para o extrato de basidiocarpo em todas as concentrações avaliadas. In vivo, os resultados indicaram o potencial de extratos de basidiocarpos de P. sanguineus para o controle de Xanthomonas axonopodis pv. phaseoli em feijoeiro, o que pode ocorrer tanto por atividade antimicrobiana direta quanto por indução de resistência envolvendo a ativação das enzimas de defesa vegetal estudadas

Made available in DSpace on 2017-07-10T17:40:49Z (GMT). No. of bitstreams: 1 Cristiane_Claudia_Meinerz.pdf: 1140657 bytes, checksum: 515f23cafb30196ea1e4641ed2debc7d (MD5) Previous issue date: 2013-08-30 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / Induced resistance involves the activation of latent defense mechanisms existing in plants in response to treatment with biotic or abiotic agents. The use of plant extracts in order to induce resistance mechanisms is an attractive alternative to chemical control, however, these extracts may occur the presence of inducer, as well ass the presence of suppressor. This study aimed to evaluate biochemical, physiological and growth of sorghum (Sorghum bicolor L.) treated with extracts of Rosmarinus officinalis, Curcuma longa and Pycnoporus sanguineus. It was evaluated the induction of phytoalexins deoxiantocianidinas, peroxidase, polyphenol oxidase, phenylalanine ammonia lyase, β-1,3 glucanase and chitinase, carbon metabolism (photosynthesis and respiration) and growth of sorghum plants. Mesocotyls sorghum were treated with the fungal and plant extracts, and acibenzolar-S-methyl (ASM) (125 mg L-1 elicitor would like reference) and distilled water. Sorghum seedlings were treated with the inducers and at 24, 48, 96 and 144 hours samples were taken for analyzes of defense enzymes. Evaluations of gas exchange were conducted periodically over 7 days with three days interval of application of foliar treatments. Were determined rate of net CO2 assimilation (A, CO2 mol m-2s-1), stomatal conductance (gs mmol H2O m-2s-1), sweating (E, H2O mmol m-2s-1) efficiency Water use (U.S. mol m-2s-1) and intrinsic efficiency of the use of water (EIUA, mol m-2s-1). For growth analysis, carried out 120 days after sowing, the parameters evaluated were: fresh leaves and roots, dry weight of leaves and roots and root volume. The parameters for production analysis were the number of panicle, seed number, mean grain mass and total production. Biotic inducers rosemary, turmeric and Pycnoporus sanguineus induced phytoalexins in mesocotyls and biochemical activities in different cultivars of sorghum, may note that the results reveal an important target for the action of elicitors in these extracts. Overall, rosemary extract caused increase in peroxidase, polyphenol oxidase and phenylalanine ammonia lyase activities, the turmeric extract induced phenylalanine ammonia-lyase activity and the extract of P. sanguineus polyphenol oxidase and β-1,3 glucanase activities. ASM interfere with the metabolism of BRS 610 sorghum in relation to the efficiency of water use (U.S.) and intrinsic efficiency of water use (EIUA), without interfering with productivity, improving the quality of production and improving markedly the number of panicle, panicle, grain number and grain weight. The fungal and plant extracts did not affect these parameters. It was possible to induce defense mechanisms in sorghum by application of extracts of rosemary, turmeric and P. sanguineus, which may allow the obtention of new molecules, and the development of alternative methods for controlling plant diseases / A indução de resistência envolve a ativação de mecanismos de defesa latentes existentes nas plantas em resposta ao tratamento com agentes bióticos ou abióticos. A aplicação de extratos vegetais visando à indução de mecanismos de resistência é uma alternativa interessante ao controle químico, entretanto, nestes extratos pode ocorrer além da presença de indutores, a presença de supressores. Este trabalho teve por objetivo avaliar aspectos bioquímicos, fisiológico e de crescimento de sorgo (Sorghum bicolor L.) tratado com extratos de Rosmarinus officinalis, Curcuma longa e Pycnoporus sanguineus. Foram avaliados a indução de fitoalexinas deoxiantocianidinas, enzimas peroxidase, polifenoloxidase, fenilalanina amônia-liase, β-1,3 glucanase e quitinase, metabolismo de carbono (fotossíntese e respiração) e crescimento de plantas de sorgo. Mesocótilos de sorgo foram tratados com os extratos vegetais e fúngico, além de acibenzolar-S-metil (ASM) (125 mg L-1 do i.a. como elicitor de referência) e água destilada. Plântulas de sorgo foram tratadas com os indutores e às 24, 48, 96 e 144 horas foram retiradas amostras para as análises das enzimas de defesa. As avaliações de trocas gasosas foram realizadas periodicamente no período de 7 dias, com três dias de intervalo da aplicação dos tratamentos via foliar. Foram determinados: taxas de assimilação líquida de CO2 (A, μmol CO2 m-2s-1), condutância estomática (gs, mmol H2O m-2s-1), transpiração (E, mmol H2O m-2s-1), eficiência do uso de água (EUA, mol m-2s-1) e eficiência intrínseca do uso de água (EIUA, mol m-2s-1). Para análise de crescimento, realizada 120 dias após a semeadura, os parâmetros avaliados foram: massa fresca das folhas e raízes; massa seca das folhas e raízes e volume de raiz. Os parâmetros avaliados na análise de produção foram: número de panícula, número de sementes; massa média de grão e produção total. Os indutores bióticos alecrim, cúrcuma e Pycnoporus sanguineus induziram fitoalexinas em mesocótilos e atividades bioquímicas nas diferentes cultivares de sorgo, podendo ressaltar que os resultados revelam um importante alvo de ação de elicitores presentes nesses extratos. De forma geral, o extrato de alecrim causou incremento na atididade de peroxidase, polifenoloxidase e fenilalanina amônia-liase; o extrato de cúrcuma induziu a atividade de fenilalanina amônia-liase e o extrato de P. sanguineus as atividades de polifenoloxidase e β-1,3 glucanase. O indutor ASM interferiu no metabolismo da cultivar BRS 610 de sorgo em relação à eficiência do uso da água (EUA) e eficiência intrínseca do uso da água (EIUA), sem interferir na produtividade, melhorando a qualidade da produção e melhorando acentuadamente o número de panícula, massa de panículas, número de grãos e massa de grãos. Os extratos vegetais e fúngico não interferiram nesses parâmetros. Foi possível induzir mecanismos de defesa em sorgo pela aplicação dos extratos de alecrim, cúrcuma e P. sanguineus, o que pode permitir a obtenção de novas moléculas e o desenvolvimento de métodos alternativos para controle de doenças em plantas

Made available in DSpace on 2017-07-10T17:36:54Z (GMT). No. of bitstreams: 1 Omari_Dangelo_Forlin_Dildey.pdf: 1490554 bytes, checksum: d179837ea4985c559fc54f5e530aaca2 (MD5) Previous issue date: 2014-02-27 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / Biotic inducing agents such as Trichoderma spp. are alternative uses for the resistance induction by activating plants mechanisms of defense. This work aimed to study the interaction of Trichoderma spp different isolates in bean plant on anthracnose control by evaluating the effects on the induction of resistance. Three experiments were conduced, one in vitro and two under greenhouse conditions. In vitro the aggressiveness of 21 Trichoderma isolates was evaluated by the culture direct confrontation method against C. lindemuthianum. In the greenhouse the bean plant was grown and treated with Trichoderma spp. (in vivo assay 1), which was inoculated via seed treatment in sterilized soil. A second assay was conducted which received inoculation from the 21 isolates inoculated via colonized rice deposition in the furrow and foliar spray of exudates of the same isolates. Leaves and roots collections for the enzyme activity determination before the exudates application, after the exudates application and collects after the inoculation were carried out. Analysis of defense enzymes (peroxidase (POX), polyphenol oxidase (PPO), fenilalanina ammonia-lyase (PAL), and β-1,3-glucanase (β-GASE)) was held. To check the endophytic colonization ability of the isolates, fragments of bean plant s roots were sanitized and placed in Petri dishes containing PDA media. The net rate of CO2 assimilation, chlorophyll content and anthracnose s severity on bean plants were also evaluated. The data were subjected to variance analysis and compared by the Scott-Knott test (p <0.05). The isolates TI2, TI4, TLB3, TLB12 (T. harzianum); TI3, TM2 (T. virens); TLB6 (T. asperellum); TLB14, TLB17 (T. koningiopsis) were considered as very efficient and the isolates TI1, TM4, TLB9, TLB15 (T. virens); TLB2, TLB4, TOD1, TOD3 (T. harzianum); TOD2A, TOD2B (T. longibrachiatum) were considered as efficient in direct confrontation against C. lindemuthianum. The isolates of Trichoderma spp, TI2 and TI4 (T. harzianum), TM4, TLB2, TLB9, TLB15, TOD1 (T. virens); TLB17 (T. koningiopsis) colonized the bean plant's roots, giving the microorganism endophytic ability on bean plant s roots. Trichoderma isolates interfere with the bean plant metabolism, acting endophytically. The isolates TI1, TM1, TLB15 (T. virens); TI2, TLB3, TLB4, TLB12, TOD1, TOD3 (T. harzianum); TLB6 (T. asperellum); TLB14, TLB17 (T. koningiopsis); TOD2B (T longibrachiatum) suppressed the β-1,3-glucanase activity in the leaf tissue, being considered as suppressors for the induction of defense on bean plants. The defense enzymes activity on the resistance induction of the bean plant treated with Trichoderma spp. for protection against C. lindemuthianum showed no major changes. The isolates did not affect the net rate of CO2 assimilation and did not change the chlorophyll content. The incidence of anthracnose's severity on bean plants, showed no significant difference by treatment via seed/soil and foliar spray of the exudates on the interaction of Trichoderma spp. evaluated. However the management with Trichoderma spp. cannot be ruled out and requires further studies in this pathossystem / Agentes indutores bióticos como Trichoderma spp. são alternativas de utilização para a indução de resistência ativando mecanismos de defesa das plantas. Este trabalho teve como objetivo estudar a interação dos diferentes isolados de Trichoderma spp. em feijoeiro no controle da antracnose, avaliando os efeitos na indução de resistência. Foram conduzidos 3 experimentos, sendo um in vitro e dois em condições de casa de vegetação. In vitro foi avaliada a agressividade de 21 isolados de Trichoderma pelo método do confronto direto de cultura contra o C. lindemuthianum. Em casa de vegetação o feijoeiro foi cultivado e tratado com isolados de Trichoderma spp. (ensaio in vivo 1), os quais foram inoculados via tratamento de semente em solo esterilizado. Um segundo ensaio foi conduzido o qual recebeu inoculação dos 21 isolados inoculados via deposição de arroz colonizado no sulco e pulverização de exsudato via foliar dos mesmos isolados. Foram realizadas coletas de folhas e raízes para determinação da atividade enzimática antes da aplicação dos exsudatos, depois da aplicação dos exsudatos e coleta após a inoculação do patógeno. Foi realizada análise das enzimas de defesa (peroxidase (POX), polifenoloxidase (PFO), fenilalanina amônia-liase (FAL) e β-1,3-glucanase (β-GASE)). Para verificar a capacidade de colonização endofítica dos isolados, fragmentos de raízes do feijoeiro foram sanitizadas e dispostas em placas de Petri contendo meio BDA. Foi avaliada também a taxa de assimilação líquida de CO2, teor de clorofila e severidade de antracnose nas plantas de feijoeiro. Os dados foram submetidos à análise de variância e comparados pelo teste de Scott-Knott (p<0,05). Os isolados TI2, TI4, TLB3, TLB12 (T. harzianum); TI3, TM2 (T. virens); TLB6 (T. asperellum); TLB14, TLB17 (T. koningiopsis) foram considerados como muito eficientes e os isolados TI1, TM4, TLB9, TLB15 (T. virens); TLB2, TLB4, TOD1, TOD3 (T. harzianum); TOD2A, TOD2B (T longibrachiatum) foram considerado como eficientes no confronto direto contra C. lindemuthianum. Os isolados de Trichoderma spp., TI2 e TI4 (T. harzianum), TM4, TLB2, TLB9, TLB15, TOD1 (T. virens); TLB17 (T. koningiopsis) colonizaram as raízes de feijoeiro, conferindo a capacidade endofítica do microrganismo nas raízes do feijoeiro. Isolados de Trichoderma interferem no metabolismo do feijoeiro, atuando endofiticamente. Os isolados TI1, TM1, TLB15 (T. virens); TI2, TLB3, TLB4, TLB12, TOD1, TOD3 (T. harzianum); TLB6 (T. asperellum); TLB14, TLB17 (T. koningiopsis); TOD2B (T longibrachiatum) suprimiram a atividade de β-1,3-glucanase no tecido foliar, sendo considerados supressores para indução da defesa das plantas de feijoeiro. A atividade das enzimas de defesa na indução de resistência de feijoeiro tratado com Trichoderma spp. para proteção a C. lindemuthianum não demonstraram grandes alterações. Os isolados não interferiram na taxa de assimilação líquida de CO2 e não alteraram o teor de clorofila. A incidência de severidade de antracnose em plantas de feijoeiro, não apresentou diferença significativa por meio do tratamento via semente/solo e pulverização dos exsudato via foliar na interação dos isolados de Trichoderma spp. avaliados. Contudo o manejo com Trichoderma spp. não pode ser descartado e requer maiores estudos nesse patossitema

Made available in DSpace on 2017-07-10T17:37:44Z (GMT). No. of bitstreams: 1 Cristiane_Claudia_Meinerz.pdf: 977726 bytes, checksum: 47602589d307d4398cc6061bcd0d1eeb (MD5) Previous issue date: 2010-02-24 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / Induction of resistance involves the activation of plant defense mechanisms in response to treatment with biotic or abiotic elicitors. The application of plant extracts in order to induce resistance mechanisms is an interesting alternative to chemical control, however, besides the presence of inducers, can occur the presence of suppressors. This study aimed to partially purificate through gel filtration chromatography (GFC) and precipitation with ammonium sulfate (SA), compounds present in decoct of Adiantum capillus-veneris, capable to induce defense mechanisms in sorghum mesocotyls, including phytoalexins and peroxidase, polyphenoloxidase (PPO), phenylalanine ammonia-lyase (PAL) and chitinase. The decoct 1% was fractionated with concentrations of ammonium sulfate, 0-20%, 20-40%, 40-60%, 60-80% and 80-100% of SA and those fractions were subjected to GFC. We obtained nine protein peaks and one glucosic peak for decoct with molecular weights ranging from 0.61 to 0.01 KDa; to fraction 0-20% were obtained two protein and two glucosic peaks, with molecular weights lower than 0.01 KDa, and concentration of sugars ranging from 4.1 to 17.5 mg mL-1; to fraction 20-40% were obtained three protein peaks (0.98 to 111.5 KDa) and five glucosic peaks (11.3 to 73.7 mg mL-1); to fraction 40-60% were obtained two protein peaks (0.09 to 111.5 KDa) and two glucosic peaks (5.6 to 7.5 mg mL-1); to fraction 60-80% were obtained six protein peaks (lower than 0.02 KDa) and two glucosic peaks (16.5 to 51.3 mg mL-1); and to fraction 80-100% were obtained three protein peaks (lower than 0.09 KDa). Sorghum mesocotyl were treated with fractions from the GFC, and decoct, acibenzolar-S-methyl (ASM) (125 mg L-1 of a. i. as elicitor of reference) and sodium phosphate buffer 10 mM pH 6.0. After incubation of 96 h were measured the levels of phytoalexins in mesocotyls and the activity of defense-related enzymes in leaves. Treatment with peak II (0,09 KDa) induced phytoalexin 6.68% more than. Among the fractionn, 60-80% increased 76% compared to ASM. To peroxidase the peak IV (lower than 0,01 KDa) increased 21% the activity compared to control water, and 44% compared to ASM. For the fraction 0-20% the protein peak II (lower than 0,01 KDa) increased 39% the activity in relation to the fraction 0-20% and 19% in relation to decoct. The fraction, 80-100% increased 89% compared to, ASM. For the PPO the peak VI (lower than 0,01 KDa) from decoct decreased 88% the activity compared to ASM. For PAL the peak II (lower than 0,01 KDa) from fraction 0-20% was 91% higher than decoct. For chitinase 1% peak IV (lower than 0,01 KDa) from decoct was 68% higher than the ASM. It was possible to induce defense mechanisms in sorghum by the application of partially purified fractions from A. capillus-veneris, which can allow to obtain new molecules and development alternative methods to control plant diseases / A indução de resistência envolve a ativação de mecanismos de defesa latentes existentes nas plantas em resposta ao tratamento com agentes bióticos ou abióticos. A aplicação de extratos vegetais visando à indução de mecanismos de resistência é uma alternativa interessante ao controle químico, entretanto, nestes extratos pode ocorrer além da presença de indutores, a presença de supressores. Este trabalho teve por objetivo a purificação parcial, por meio de cromatografia de filtração em gel (CFG) e precipitação com sulfato de amônio (SA), de compostos presentes em decocto de avenca (Adiantum capillus-veneris), eficientes na indução de mecanismos de defesa em mesocótilos de sorgo, incluindo as fitoalexina deoxiantocianidinas e as proteínas peroxidase, polifenoloxidase, fenilalanina amônia-liase e quitinase, buscando selecionar frações potencialmente eficientes na indução de resistência em plantas. Decocto (EA 1%) de A. capillus-veneris foi fracionado com concentrações de sulfato de amônio de 0-20%, 20-40%, 40-60%, 60-80% e 80-100% e esses cortes foram submetidos à cromatografia de filtração em gel (CFG). Foram obtidos nove picos protéicos e um pico glicídico para EA 1% com massas moleculares variando de 0,61 à 0,01 KDa; no corte 0-20% foram obtidos dois picos protéicos e dois glicídicos, com massas moleculares menores que 0,01 KDa, e concentração de açúcares redutores variando de 4,1 a 17,5 µg mL-1; no corte 20-40% três picos protéicos (111,5 à 0,98 KDa) e cinco glicídicos (11,3 a 73,7 µg mL-1 de açúcares); no corte 40-60% dois picos protéicos (111,5 à 0,09 KDa) e dois glicídicos (5,6 a 7,5 µg mL-1); no corte 60-80% seis picos protéicos (menor que 0,02 KDa) e dois glicídicos (16,5 a 51,3 µg mL-1); e no corte 80-100% três picos protéicos (menor que 0,09 KDa). Mesocótilos de sorgo foram tratados com as frações provenientes da CFG, além do decocto a 1%, acibenzolar-S-metil (ASM) (125 mg. L-1 do i.a. como elicitor de referência) e tampão fosfato de sódio 10 mM pH 6,0, totalizando 42 tratamentos. Após incubação por um período de 96 h, avaliou-se dos teores de fitoalexinas nos mesocótilos e análises bioquímicas dos folíolos. O tratamento pico II (0,09 KDa) do EA 1% mostrou-se eficiente na indução de fitoalexinas, sendo superior em 6,68% ao ASM. Entre os cortes, 60-80% permitiu incremento de 76% em relação ao ASM. Para peroxidase o pico IV (menor que 0,01 KDa) do EA 1% incrementou 21% a atividade em relação a testemunha água e 44% ao ASM. Para os precipitados 0-20% o pico protéico II (menor que 0,01 KDa) promoveu incremento de 39% na atividade em relação ao corte 0-20% e 19% para o EA 1%. O precipitado 80-100% foi superior 89% ao ASM. Para polifenoloxidase o pico protéico VI (menor que 0,01 KDa) do EA1% reduziu 88% a atividade em relação ao ASM. Para fenilalanina amônia-liase o pico protéico II (menor que 0,01 KDa) do corte 0-20% foi 91% superior ao EA 1%. Para quitinase o pico protéico IV (menor que 0,01 KDa) do EA 1% foi 68% superior ao ASM. Foi possível induzir mecanismos de defesa em sorgo pela aplicação de frações parcialmente purificadas de A. capillus-veneris, o que pode permitir a obtenção de novas moléculas e o desenvolvimento de métodos alternativos para controle de doenças em plantas

Les phytoplasmes sont des bactéries phytopathogènes, sans paroi, qui appartenant à la classe des Mollicutes. Ils ne peuvent pas etre cultivés in vitro et sont limités à des tubes du phloème. Ils provoquent des centaines de maladies chez de nombreuses espèces végétales dans le monde entier, ce qui conduit à des pertes de récolte importantes. Les phytoplasmes sont transmis naturellement par des insectes suceurs de sève dans laquelle ils se multiplient. Ils induisent des symptômes graves, notamment le jaunissement, la croissance limitée, déclin, ainsi que des anomalies des fleurs et des fruits. L'infection par le phytoplasme du stolbur, en particulier, affect fortement la morphologie florale. Dans la tomate, deux isolats différents du phytoplasme du stolbur, nommé C et PO, induisent des symptômes différents. La tomate infectée par le phytoplasme du stolbur PO montrent des malformations florale telles que les sépales hypertrophiés, les pétales et les étamines avortées ce qui conduit à la stérilité. En revanche, la tomate infecté par le phytoplasme du stolbur C ont de petites feuilles de tomate en retrait, mais les fleurs presque normale, et produisent des fruits. Nous avons précédemment montré que SlDEF, un gène impliqué dans la formation des pétales est réprimé dans des plantes de tomate infectée par le stolbur phytoplasme PO. Toutefois, l'expression de son facteur de transcription, codée par le gène FA, est resté stable ou voir légèrement augmentée. Nous avons donc émis l'hypothèse que la répression de SlDEF pourrait être dû à une méthylation de l'ADN. Pour tester cette hypothèse, nous avons étudié l'expression des gènes de méthylases et de déméthylases. Ils étaient en général réprimés dans les tomates infectées par le phytoplasme du stolbur PO, ce qui était en accord avec l'hypothèse De plus, nous avons étudié les voies de défense activée chez les tomates infectées par le phytoplasme du stolbur. Pour se défendre, les plantes utilisées des molécules de signalisation comme l'acide salicylique (SA), l'acide jasmonique (JA) et d'éthylène (ET). Nous avons étudié l'expression de 21 gènes de défense dépendants SA / JA / ET, des gènes de biosynthèse et les facteurs de transcription chez les tomates infectées par les phytoplasmes du stolbur C et PO. Nous avons également étudié l'effet de la pré-activation des voies de SA et JA sur la production des symptômes. Nos résultats montrent clairement que les voies de défense ont été activées différemment dans les tomates infectés par le phytoplasme du stolbur C et PO. En effet, les voies de défense dépendantes de SA, ET et JA ont été activées chez les tomates infectées par le phytoplasme du stolbur C alors que seulement les voies dépendantes SA et ET ont été activés dans les tomates infectées par stolbur PO . En outre, la pré-activation de la voie de défense dépendante SA par l'application de BTH modifie légèrement l'évolution des symptômes de maladies causées par le phytoplasme du stolbur PO / Phytoplasma are cell wall-less, phytopathogenic bacteria belonging to the class Mollicutes. They have not been cultured in vitro and are restricted to the phloem sieve tubes. They cause hundreds of diseases in many plant species worldwide, resulting in important crop losses. Phytoplasmas are naturally transmitted by sap-sucking insects in which they multiply. They induce severe symptoms including yellowing, restricted growth, decline, as well as major flowers and fruits abnormalities.The stolbur phytoplasma infection, in particular, has been reported to strongly affect floral morphology. In tomato, two different isolates of stolbur phytoplasma, named C and PO, induce different symptoms. The stolbur PO phytoplasma-infected plants show abnormal flower development such as hypertrophied sepals, and aborted petals and stamens leading to sterility. In contrast, stolbur C phytoplasma-infected tomato have small indented leaves but nearly normal flowers, and produce fruits. We have previously shown that SlDEF, one gene involved in petal formation, was repressed in stolbur PO phytoplasma-infected tomato. However, the expression of its transcription factor, encoded by the gene FA, was unchanged or slightly up-regulated. So we hypothesized that SlDEF repression could be due to DNA methylation. To test this hypothesis, we studied the expression of DNA methylases and demethylases genes. They were in general down-regulated in stolbur PO infected tomato, which was in agreement with the hypothesis. However, the regulation of SlDEF expression could not be firmly correlated to the DNA methylation status of its promoter region. In addition, we studied the plant defense pathways activated in stolbur phytoplasma-infected tomato. To defend themselves, plants used signalling molecules like Salicylic acid (SA), Jasmonic acid (JA) and Ethylene (ET). We studied the expression of 21 SA/JA/ET regulated defense and biosynthesis genes including transcription factors in stolbur C and PO phytoplasma-infected tomato as compared to healthy ones. We also studied the effect of pre-activation of SA and JA mediated defense pathways on symptom production. Our results clearly showed that defense pathways were activated differently in stolbur C and PO phytoplasma-infected tomato. Indeed, SA ET and JA dependant pathways were activated in stolbur C-infected tomato while only SA and ET dependant pathways were activated in stolbur PO-infected plants. In addition, pre-activation of SA-dependent defense pathway by application of BTH slightly modify the evolution of disease symptoms caused by stolbur PO phytoplasma whereas no effect was observed after treatment with an analogue of JA.