Memoria para optar al Título Profesional de: Ingeniero Agrónomo / El control químico para la pudrición gris es la principal herramienta para reducir su incidencia que puede ser lograda por medio de diversos compuestos fungicidas. El fludioxonil, ciprodinil y fenhexamid son moléculas con actividad botriticida y fueron desarrolladas a partir de los años 90. Sin embargo su eficacia se ha visto fuertemente afectada por la aparición de resistencia debido a su uso reiterado. Por otro lado algunos ingredientes activos utilizados en determinadas concentraciones suelen alterar el metabolismo de las plantas. El objetivo de este estudio fue evaluar el efecto de dos fungicidas aplicados al follaje de plantas jóvenes de Vitis vinifera, variedades Thompson Seedless y Chardonay sobre la actividad fotosintética. Las mediciones fueron realizadas 5 veces a lo largo del día durante 5 días. Se evaluaron los parámetros de fotoinhibición de la fotosíntesis, contenido de clorofilas, tasa de asimilación de CO2 y permeabilidad relativa de membranas. Para ello se realizaron dos ensayos consistentes en 4 tratamientos más un testigo (T1= ciprodinil (375g/kg) & fludioxonil (250g/kg), 60gr/hL; T2= ciprodinil (375g/kg) & fludioxonil (250g/kg), 120gr/hL agua; T3= fenhexamid (500g/L) 72,5cc/hL; T4= fenhexamid (500g/L) 145cc/hL agua). Los resultados no mostraron diferencias significativas entre tratamientos a lo largo del ensayo, indicando que los fungicidas no alteraron los parámetros fisiológicos registrados de las plantas. / Chemical control for gray mold is the principal tool to reduce its occurrence and it can be achieved with a number of different fungicide compounds. Fludioxonil, ciprodinil and fenhexamid are molecules with botriticide activity and were developed from the ’90s. However their efficacy has been seen to be markedly affected by the development of resistance due to repeated use. Moreover some active ingredients used in certain concentrations often alter plants’ metabolism. The aim of this study was to assess the effects of two fungicides applied to the leaves of young plants from Vitis vinifera varieties Thompson Seedless and Chardonnay on their photosynthetic activity. The measurements were carried out five times a day for five days. The variables that were tested were photoinhibition of photosynthesis; Chlorophylls content; Carbon dioxide assimilation and Membrane permeability. Two experiments were performed, consisting of four treatments and a control (T1= ciprodinil (375g/kg) & fludioxonil (250g/kg), 60gr/hL; T2= ciprodinil (375g/kg) & fludioxonil (250g/kg), 120gr/hL agua; T3= fenhexamid (500g/L) 72,5cc/hL; T4= fenhexamid (500g/L) 145cc/hL agua). The results did not show differences between plants subject to the different treatments over the course of the experiments, suggesting that fungicides do not affect the plants physiology.

Memoria para optar al título profesional de: Ingeniero Agrónomo Mención: Sanidad Vegetal / La pudrición gris, causada por el hongo Botrytis cinerea, corresponde al principal problema fitopatológico que enfrentan los productores de uva de mesa, puesto que limita su producción y exportación al desarrollarse pudriciones incluso en almacenaje. Para su control, se integran diferentes medidas entre las cuales el uso de fungicidas específicos es la base de los programas utilizados. Actualmente, la creciente preocupación por la presencia de residuos de fungicidas en la fruta y el riesgo ambiental asociado a su uso, además del desarrollo de resistencias en Botrytis a estas moléculas, han complicado su empleo como estrategia de control. Ante el cual se han aplicado métodos alternativos como la intensificación de prácticas culturales que disminuyan las condiciones predisponentes de la enfermedad, junto a herramientas naturales que incluyen el uso de extractos de cítricos y controladores biológicos como Trichoderma. Por otro lado, recientemente se ha elaborado un biofungicida con la bacteria antagonista Serratia plymuthica cepa CCGG2742, un controlador biológico de B. cinerea, que en este ensayo se ha probado su sensibilidad frente a fungicidas de uso común en vid, con el objeto de conocer su compatibilidad en un posible programa de control que integre estas estrategias. Se evaluaron los ingredientes activos boscalid, ciprodinil + fludioxonil, fenhexamida, iprodione, kresoxim methyl, pyrimethanil, tebuconazole y extracto de cítrico en concentraciones que fluctuaron entre 0 y 5.000 µg i.a. ∙ mL-1 y se calculó la EC50. Los valores de EC50 obtenidos fueron: 6,00∙104 ; 3,18∙107 ; 7,04∙1012 y 1,38∙1018 µg i.a. ∙ mL-1 , para fenhexamida, extracto de cítrico, tebuconazole y boscalid respectivamente, mientras que con iprodione, kresoxim methyl, pyrimethanil y la mezcla de c+f, no se obtuvieron valores de EC50 positivos, ni indicios de inhibición in vitro. De acuerdo a estos resultados, el uso de dosis comerciales de fungicidas no altera el desarrollo de la cepa CCGG2742. / Gray mold induced by Botrytis cinerea, is the most important disease of table grapes that causes significant losses to grape growers. Its ability to attack in the orchard and its ability to develop under conditions prevailing during storage, shipment and marketing make its control a challenge. Control programs have relied mainly on chemical strategies using specific fungicides. However, the growing public concern about fungicide residues in fruit, and the environmental risk associated to fungicide use, in addition to pathogen’s resistance development, have created a complicated situation for the continuous use of fungicides. Therefore, alternative methods have been developed for non-chemical control, such as cultural practices, and the use of natural products including citrus extracts and biological controls agents, as Trichoderma. Serratia plymuthica, antagonistic to B. cinerea, is a new biocontrol agent, and in this study the sensitivity of the strain CCGG2742 to fungicides commonly used in vineyards was tested, in order to know their compatibility for possible control programs that integrate these tools. Boscalid, ciprodinil + fludioxonil, fenhexamida, iprodione, kresoxim methyl, pyrimethanil, tebuconazole and citric extract were tested at concentrations ranging between 0 and 5,000 µg i.a. ∙ mL-1 . EC50 values obtained ranged between 5.48∙104 and 1.38∙1018 µg i.a. ∙ mL-1 . According to these results, commercial rates of the fungicides tested do not affect the development of S. plymuthica strain CCGG2742.

Submitted by VANESSA VOIGT PALARETTI (van_voigt@yahoo.com.br) on 2018-06-14T01:09:15Z No. of bitstreams: 1 TESE.vanessapalaretti.pdf: 4088105 bytes, checksum: 442eb1364fba0bb29cde74e7b9f5b3f1 (MD5) / Approved for entry into archive by Neli Silvia Pereira null (nelisps@fcav.unesp.br) on 2018-06-14T12:10:14Z (GMT) No. of bitstreams: 1 palaretti_vv_dr_jabo_int.pdf: 4088105 bytes, checksum: 442eb1364fba0bb29cde74e7b9f5b3f1 (MD5) / Made available in DSpace on 2018-06-14T12:10:14Z (GMT). No. of bitstreams: 1 palaretti_vv_dr_jabo_int.pdf: 4088105 bytes, checksum: 442eb1364fba0bb29cde74e7b9f5b3f1 (MD5) Previous issue date: 2018-05-04 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / As doenças fúngicas são as principais causas de perdas na pós-colheita do tomate, e são controladas por manejo cultural e fungicidas aplicados na pré-colheita. O tratamento hiperbárico na pós-colheita é um métodos físico, com potencial uso comercial na redução da deteriorização por fungos. Neste trabalho, a eficiência das pressões hiperbáricas para controlar o desenvolvimento das podridões de Botrytis cinerea e Alternaria sp. in vitro e em tomates ‘Débora’ foram avaliadas, bem como a atividade das enzimas antioxidantes e as relacionadas a defesa do vegetal. Os experimentos foram realizados em um sistema de pressão hiperbárica do Laboratório de Tecnologia Pós-colheita da FCAV-UNESP, Câmpus de Jaboticabal, utilizando as pressões de 100 (controle), 200, 400, 600 e 800 kPa à temperatura de 23±1°C, durante 3 e 6 dias. No ensaio in vitro as placas com os fungos permaneceram nas pressões e depois foram incubadas até que a colônia do controle atingisse a borda. No experimento in vivo, tomates inoculados com os fungos foram mantidos por 3 e 6 dias sob o tratamento nas pressões, com posterior armazenamento por mais 2 e 4 dias em condição de ambiente (23 °C, 50% UR, 100 kPa) para simular a comercialização. Avaliou-se o crescimento micelial, a taxa de germinação dos conídios dos fungos, incidência e severidade das podridões nos frutos, o metabolismo antioxidante (teor de licopeno e polifenóis totais) e a atividade das enzimas relacionadas à defesa dos tomates (peroxidase (POD), fenilalanina-amônia-liase (PAL), β 1,3 glucanase (GLU), superóxido dismutase (SOD) e catalase (CAT). As pressões hiperbáricas não apresentaram efeito fungistático sobre os patógenos, entretanto, houve redução no crescimento micelial e na taxa de germinação de B. cinerea após 3 dias na pressão de 800 kPa. As pressões elevadas de 600 e 800 kPa durante 3 dias reduziram a incidência (25%) e a severidade do mofo cinzento (68%), após 4 dias de armazenamento (3+4 dias). Enquanto a incidência e severidade da mancha de Alternaria (Alternaria sp.) não tiveram redução expressiva em ambos os períodos. Na pressão de 800 kPa, os frutos inoculados com B. cinerea e Alternaria sp. apresentaram um retardo no amadurecimento, evidenciado pelo menor conteúdo de licopeno e menor atividade da enzima POD, a qual está relacionada à senescência dos frutos. As pressões elevadas (600 e 800 kPa) promoveram a atividade das enzimas PAL, GLU e CAT dos tomates inoculados de ambos os patógenos por 3 dias, mantendo tendência similar após as condições de ambiente. Os frutos tratados com as pressões elevadas apresentaram incremento no conteúdo de polifenóis totais em relação ao controle, durante os períodos de armazenamento. As pressões não influenciaram a atividade da enzima SOD dos tomates, após os períodos de tratamentos. As pressões hiperbáricas de 600 e 800 kPa foram eficientes na redução de doenças na pós-colheita de tomates e também podem ter induzido a atividade de enzimas envolvidas na resistência às doenças. / Fungal rots are one of the major causes of postharvest losses in tomatoes, and they are controlled by cultural management and fungicidal applied in preharvest. The postharvest hyperbaric treatment is a physical method that it have potential to be used in commercial for reducing fungal decay. In this work, the efficiency of hyperbaric pressures to control rot development of Botrytis cinerea and Alternaria sp. in vitro and ‘Debora’ tomatoes were evaluated, also the activity of antioxidant and defense-related enzymes.The experiments were carried out in a hyperbaric pressure system at FCAV-UNESP Postharvest Technology Laboratory, Campus de Jaboticabal, using the pressures of 100 (control), 200, 400, 600 and 800 kPa at a temperature of 23 ± 1 ° C for 3 and 6 days. In vitro assay the Petri dishes with the fungi were kept at pressures after they were incubated until the control colony reached the edge. In vivo assay, tomatoes inoculated with the fungus were kept at 3 and 6 days under the treatment pressures, after they were stored for more 2 and 4 days at ambient conditions (23 °C, 50% RH, 100 kPa) to simulate the commercialization. It was evaluated the mycelial growth, germination rate of fungal conidia, incidence and severity of fruit rot, antioxidant metabolism (lycopene content and total polyphenols) and activity of defence-related enzymes (eroxidase (POD), phenylalanine ammonia-lyase (PAL), β 1,3 glucanase (GLU), superoxide dismutase (SOD) and catalase (CAT)). The hyperbaric pressures had no fungistatic effect on the pathogens, however, there was a reduction in mycelial growth and in the germination rate of B. cinerea after 3 days at 800 kPa. High pressures of 600 and 800 kPa for 3 days reduced the incidence (25%) and gray mold severity (68%) after 4 days of the storage (3 + 4 days). While the incidence and severity of black rot (Alternaria sp.) didn´t decrease significantly in both periods. At the pressure of 800 kPa, the fruits inoculated with B. cinerea and Alternaria sp. demonstrated a delay in ripening, as evidenced by the lower lycopene content and lower activity of the POD enzyme, which is related to fruit senescence. The high pressures (600 and 800 kPa) promoted the activity of the PAL, GLU and CAT enzymes of the tomatoes inoculated from both pathogens for 3 days, maintaining a similar trend after the ambient conditions. The fruits treated with high pressures demonstrated an increase in the total polyphenols content in relation to the control during storage periods. The pressures didn´t influence the activity of the SOD enzyme of the tomatoes, after the periods of treatments. Hyperbaric pressures of 600 and 800 kPa were efficient in reducing diseases in postharvest tomatoes and they must have induced the activity of enzymes involved in resistance to diseases.

Diversos estudos vêm demonstrando o potencial de plantas medicinais no controle de fitopatógenos, tanto por sua ação fungistática direta, quanto pela capacidade de induzir a defesa das plantas, indicando a presença de moléculas com características elicitoras. Nesse sentido foram desenvolvidos três experimentos no ano de 2009 e 2010, na Universidade Tecnológica Federal do Paraná – Campus de Dois Vizinhos, com objetivos de avaliar o potencial de preparados a base de calêndula (Calendula officinalis L.) na indução de fitoalexinas em cotilédones de soja, na indução de resistência em frutos de morango e o efeito fungistático sobre Botrytis cinerea in vitro. O delineamento experimental utilizado para os experimentos foi inteiramente casualizado com 15 tratamentos resultantes da combinação de três formas de extração (extrato alcoólico, infusão e maceração) e cinco concentrações (zero; 1,25; 2,5; 5 e 10%), sendo a concentração zero água destilada, em arranjo fatorial (3 x 5) com 4 repetições. O primeiro experimento avaliou o potencial fungistático dos preparados sobre B. cinerea in vitro. Em placas de Petri® foram adicionados no meio de cultura BDA (Batata Dextrose Ágar) os preparados nas diferentes concentrações. Após a solidificação, um orifício de 8 mm foi realizado no centro da placa e introduzindo-se 2μL da suspensão de conídios de B. cinerea. As placas foram mantidas em câmara de crescimento durante 7 dias a 25ºC, e no sétimo dia mediu-se o diâmetro do halo de crescimento do fungo. O segundo experimento avaliou a indução de fitoalexinas em cotilédones de soja em resposta aos preparados à base de calêndula. Sementes de soja foram semeadas em areia autoclavada e mantidas em temperatura ambiente por 10 dias. Em seguida os cotilédones das plântulas foram removidos e na face abaxial destes foram aplicados os tratamentos. Após seguir os procedimentos metodológicos da técnica de extração, obteve-se via espectrofotometria a quantificação da fitoalexina gliceolina. O terceiro experimento avaliou o efeito dos preparados a base de C. officinalis sobre os parâmetros físico-químicos e bioquímicos de frutos de morango relacionados com a indução de resistência. Os frutos foram acondicionados em bandejas plásticas e pulverizados com os diferentes tratamentos. Após 6 horas da pulverização dos preparados foi pulverizada solução contendo cerca 104 conídios do fungo B. cinerea. As avaliações foram realizadas após 3 dias da implantação do experimento. Os parâmetros avaliados foram: perda de massa, incidência de podridões, acidez titulável, firmeza de polpa e sólidos solúveis totais, e as bioquímicas foram: açúcares totais, antocianinas, flavonóides e atividade da enzima fenilalanina amônia-liase (FAL). Os resultados demonstraram que a maceração em todas as suas concentrações inibiu o crescimento do fungo B. cinerea in vitro, sendo que a partir de 2,5% observou-se inibição total. O tratamento com infusão na sua maior concentração (10%) também apresentou resposta positiva na inibição do crescimento de B. cinerea. Os preparados de C. officinalis apresentaram capacidade de indução das fitoalexinas gliceolinas em cotilédones de soja. Na aplicação dos preparados em pós-colheita de morangos, não ocorreu influência dos tratamentos sobre perda de massa, incidência de podridões, firmeza de polpa, acidez titulável, sólidos solúveis totais, açúcares totais e antocianina. Para flavonóides o extrato alcoólico em todas as concentrações e a infusão a partir de 5% estimularam sua produção. A atividade da enzima FAL foi estimulada pela aplicação dos extratos. / Several studies have demonstrated the potential of medicinal plants in the pathogens control, through its direct fungistatic action or its ability to induce plant defense, it suggesting the presence of molecules with characteristics elicitors. Three experiments were carried out on 2009 and 2010, in the Universidade Tecnológica Federal do Paraná, Dois Vizinhos city (PR – Brazil). The aim of this work was to evaluate prepared caledula (Calendula officinalis L.) potential for phytoalexins induction in the soybean cotyledons, resistance induction in the strawberry fruits and fungistatic effect of Botrytis cinerea in vitro. The experimental design was completely randomized with 15 treatments resulting from the combination of three forms of extraction (alcohol extract, infusion and maceration) and five concentrations (zero, 1.25, 2.5, 5 and 10%), with zero concentration distilled water in a factorial (3 x 5) with four replications the distilled water was used as control. The first experiment evaluated the potential fungistatic preparations in the B. cinerea control. The preparations with the different concentrations were put in Petri dishes with BDA culture media. After the media solidification, a hole of 8 mm was punched in the dishes Petri center and in this hole was introduced 2μL of spore B. cinerea suspension. The Petri dishes were kept in growth chamber for 7 days at 25°C. In the seventh day, the halo diameter fungus growth was evaluated. The second experiment evaluated the phytoalexins induction in the soybean cotyledons soybean as result of C. officinalis derivatives. Soybean seeds were sown in sterilized sand and it were kept at room temperature during 10 days. Then, the seedlings cotyledons were removed and on the abaxial part it was applied the treatments. After methodological procedures of extraction technique, it was obtained the phytoalexins glyceolin quantification by for spectrophotometry. The third study evaluated the C. officinalis solutions effect on the strawberry fruits physic-chemical and biochemical characteristics. The fruits were packed in plastic trays and it sprayed with different treatments. After six hours of the treatments spraying, it was sprayed a solution containing about 10-4 spore of B. cinerea fungus. After 3 days, it was realized the evaluations. The mass loss, rots incidence, titratable acidity, flesh firmness, total soluble solids, totals sugars, anthocyanins, flavonoids and activity of phenylalanine-amonialiase (PAL) were evaluated. The results showed that the maceration with all over the concentrations used, it inhibited of the B. cinerea fungus in vitro growth. Concentrations higher that 2.5%, presented total inhibition ability. Treatment with infusion using the highest concentration (10%) also showed positive response for B. cinerea growth inhibiting. The C. officinalis solutions presented induction capacity of the phytoalexins glyceolin in soybean cotyledons. It was observed that the solutions applied during the strawberry postharvest didn’t influence in the physico-chemical characteristics (mass loss, rots incidence, titratable acidity, flesh firmness and total soluble solids). In the biochemical analyzes for totals sugars and anthocyanins wasn’t observed statistical difference among the treatments. The alcoholic extract all over the concentrations and the infusion with concentrations higher than 5% stimulated the flavonoids productions. The PAL enzyme activity was stimulated by the application of extracts.

Diversos estudos vêm demonstrando o potencial de plantas medicinais no controle de fitopatógenos, tanto por sua ação fungistática direta, quanto pela capacidade de induzir a defesa das plantas, indicando a presença de moléculas com características elicitoras. Nesse sentido foram desenvolvidos três experimentos no ano de 2009 e 2010, na Universidade Tecnológica Federal do Paraná – Campus de Dois Vizinhos, com objetivos de avaliar o potencial de preparados a base de calêndula (Calendula officinalis L.) na indução de fitoalexinas em cotilédones de soja, na indução de resistência em frutos de morango e o efeito fungistático sobre Botrytis cinerea in vitro. O delineamento experimental utilizado para os experimentos foi inteiramente casualizado com 15 tratamentos resultantes da combinação de três formas de extração (extrato alcoólico, infusão e maceração) e cinco concentrações (zero; 1,25; 2,5; 5 e 10%), sendo a concentração zero água destilada, em arranjo fatorial (3 x 5) com 4 repetições. O primeiro experimento avaliou o potencial fungistático dos preparados sobre B. cinerea in vitro. Em placas de Petri® foram adicionados no meio de cultura BDA (Batata Dextrose Ágar) os preparados nas diferentes concentrações. Após a solidificação, um orifício de 8 mm foi realizado no centro da placa e introduzindo-se 2μL da suspensão de conídios de B. cinerea. As placas foram mantidas em câmara de crescimento durante 7 dias a 25ºC, e no sétimo dia mediu-se o diâmetro do halo de crescimento do fungo. O segundo experimento avaliou a indução de fitoalexinas em cotilédones de soja em resposta aos preparados à base de calêndula. Sementes de soja foram semeadas em areia autoclavada e mantidas em temperatura ambiente por 10 dias. Em seguida os cotilédones das plântulas foram removidos e na face abaxial destes foram aplicados os tratamentos. Após seguir os procedimentos metodológicos da técnica de extração, obteve-se via espectrofotometria a quantificação da fitoalexina gliceolina. O terceiro experimento avaliou o efeito dos preparados a base de C. officinalis sobre os parâmetros físico-químicos e bioquímicos de frutos de morango relacionados com a indução de resistência. Os frutos foram acondicionados em bandejas plásticas e pulverizados com os diferentes tratamentos. Após 6 horas da pulverização dos preparados foi pulverizada solução contendo cerca 104 conídios do fungo B. cinerea. As avaliações foram realizadas após 3 dias da implantação do experimento. Os parâmetros avaliados foram: perda de massa, incidência de podridões, acidez titulável, firmeza de polpa e sólidos solúveis totais, e as bioquímicas foram: açúcares totais, antocianinas, flavonóides e atividade da enzima fenilalanina amônia-liase (FAL). Os resultados demonstraram que a maceração em todas as suas concentrações inibiu o crescimento do fungo B. cinerea in vitro, sendo que a partir de 2,5% observou-se inibição total. O tratamento com infusão na sua maior concentração (10%) também apresentou resposta positiva na inibição do crescimento de B. cinerea. Os preparados de C. officinalis apresentaram capacidade de indução das fitoalexinas gliceolinas em cotilédones de soja. Na aplicação dos preparados em pós-colheita de morangos, não ocorreu influência dos tratamentos sobre perda de massa, incidência de podridões, firmeza de polpa, acidez titulável, sólidos solúveis totais, açúcares totais e antocianina. Para flavonóides o extrato alcoólico em todas as concentrações e a infusão a partir de 5% estimularam sua produção. A atividade da enzima FAL foi estimulada pela aplicação dos extratos. / Several studies have demonstrated the potential of medicinal plants in the pathogens control, through its direct fungistatic action or its ability to induce plant defense, it suggesting the presence of molecules with characteristics elicitors. Three experiments were carried out on 2009 and 2010, in the Universidade Tecnológica Federal do Paraná, Dois Vizinhos city (PR – Brazil). The aim of this work was to evaluate prepared caledula (Calendula officinalis L.) potential for phytoalexins induction in the soybean cotyledons, resistance induction in the strawberry fruits and fungistatic effect of Botrytis cinerea in vitro. The experimental design was completely randomized with 15 treatments resulting from the combination of three forms of extraction (alcohol extract, infusion and maceration) and five concentrations (zero, 1.25, 2.5, 5 and 10%), with zero concentration distilled water in a factorial (3 x 5) with four replications the distilled water was used as control. The first experiment evaluated the potential fungistatic preparations in the B. cinerea control. The preparations with the different concentrations were put in Petri dishes with BDA culture media. After the media solidification, a hole of 8 mm was punched in the dishes Petri center and in this hole was introduced 2μL of spore B. cinerea suspension. The Petri dishes were kept in growth chamber for 7 days at 25°C. In the seventh day, the halo diameter fungus growth was evaluated. The second experiment evaluated the phytoalexins induction in the soybean cotyledons soybean as result of C. officinalis derivatives. Soybean seeds were sown in sterilized sand and it were kept at room temperature during 10 days. Then, the seedlings cotyledons were removed and on the abaxial part it was applied the treatments. After methodological procedures of extraction technique, it was obtained the phytoalexins glyceolin quantification by for spectrophotometry. The third study evaluated the C. officinalis solutions effect on the strawberry fruits physic-chemical and biochemical characteristics. The fruits were packed in plastic trays and it sprayed with different treatments. After six hours of the treatments spraying, it was sprayed a solution containing about 10-4 spore of B. cinerea fungus. After 3 days, it was realized the evaluations. The mass loss, rots incidence, titratable acidity, flesh firmness, total soluble solids, totals sugars, anthocyanins, flavonoids and activity of phenylalanine-amonialiase (PAL) were evaluated. The results showed that the maceration with all over the concentrations used, it inhibited of the B. cinerea fungus in vitro growth. Concentrations higher that 2.5%, presented total inhibition ability. Treatment with infusion using the highest concentration (10%) also showed positive response for B. cinerea growth inhibiting. The C. officinalis solutions presented induction capacity of the phytoalexins glyceolin in soybean cotyledons. It was observed that the solutions applied during the strawberry postharvest didn’t influence in the physico-chemical characteristics (mass loss, rots incidence, titratable acidity, flesh firmness and total soluble solids). In the biochemical analyzes for totals sugars and anthocyanins wasn’t observed statistical difference among the treatments. The alcoholic extract all over the concentrations and the infusion with concentrations higher than 5% stimulated the flavonoids productions. The PAL enzyme activity was stimulated by the application of extracts.

Submitted by Marco Antônio de Ramos Chagas (mchagas@ufv.br) on 2018-09-19T17:34:45Z No. of bitstreams: 1 texto completo.pdf: 672894 bytes, checksum: 1244927f20e5eed71f3171916884e265 (MD5) / Made available in DSpace on 2018-09-19T17:34:45Z (GMT). No. of bitstreams: 1 texto completo.pdf: 672894 bytes, checksum: 1244927f20e5eed71f3171916884e265 (MD5) Previous issue date: 2016-02-24 / Conselho Nacional de Desenvolvimento Científico e Tecnológico / Botrytis cinerea, o agente etiológico do mofo cinzento, é um patógeno importante em morangueiros. Clonostachys rosea é efetivo no biocontrole dessa doença em diversas culturas, em condições controladas e de campo. O antagonista pode, ainda, promover o crescimento, aumentar a produção e induzir respostas de defesa nas plantas. Objetivando-se investigar os efeitos da aplicação do antagonista na população do patógeno e no metabolismo da planta, estudou-se a interação entre C. rosea, B. cinerea e morangueiros com base na dinâmica populacional dos fungos e no perfil metabólico do hospedeiro. Avaliou-se a colonização foliar de ambos os fungos e a sobrevivência epifítica de C. rosea em folhas, em câmara de crescimento e em cultivos comerciais. O antagonista se estabeleceu e reduziu a colonização por B. cinerea, principalmente quando aplicado semanalmente. A população de C. rosea diminuiu com o tempo após a aplicação, e a intensidade de colonização pelo patógeno foi inversamente proporcional à do antagonista. Em câmara de crescimento, a população epifítica do antagonista reduziu-se substancialmente aos 3 dias da aplicação. Em condições de campo, onde o intervalo mínimo entre a última aplicação e coleta de folhas foi de 7 dias, não foi possível quantificar a população epifítica do antagonista. Comparou-se o perfil metabólico de morangueiros tratados com C. rosea em intervalos de 7, 14 ou 28 dias ao de morangueiros não tratados, e verificou-se que a aplicação semanal alterou o perfil metabólico. Aumentos significativos nos níveis de citrato e succinato, compostos intermediários do ciclo de Krebs, de alguns fitoesteróis, fitoestrogênios, cafeína e piceatanol foram observados. Tais alterações podem estar relacionadas à promoção de crescimento e ativação do sistema de defesa. Concluiu-se que C. rosea afetou a dinâmica populacional de B. cinerea em tecidos assintomáticos e alterou o perfil metabólico de morangueiros. Esses resultados subsidiam estudos subsequentes para elucidar o modo de ação de agentes de biocontrole. / Botrytis cinerea, the causal agent of gray mold, is an important pathogen of strawberry plants. Clonostachys rosea is effective in the biocontrol of gray mold in several crops, in growth chamber or field conditions. The antagonist may also promote growth, increase the production and induce defense responses of plants. Aiming to investigate the effects of antagonist application on pathogen population and plant metabolism, we studied the interaction of C. rosea, B. cinerea and strawberry plants based on the population dynamics of both fungi and metabolic profile of the host. We assessed leaf colonization of both fungi and the epiphytic survival of C. rosea on leaves, under growth chamber and in commercial cultivations. The antagonist established in leaves and reduced the colonization by B. cinerea, mainly when applied weekly. The population of C. rosea decreased over time after the application and tissue colonization by the pathogen was inversely proportional to the colonization by the antagonist. Under growth chamber the antagonist population was substantially reduced 3 days after applied. Under field conditions, which the minimal interval between last application and the collection of leaves was 7 days, it was not possible to quantify the epiphytic population of the antagonist. We compared the metabolic profiles of strawberry plants treated with C. rosea at intervals of 7, 14, or 28 days with the profiles of untreated plants, and found that weekly applications altered the metabolic profiles. There was a significant increase in the levels of citrate and succinate, intermediate compounds in the Krebs cycle, some phytosterols, phytoestrogens, caffeine, and piceatannol. These changes may be related to growth promotion and activation of plant defense system. In conclusion, C. rosea affected the population dynamics of B. cinerea in symptomless tissues and changed the metabolic profile of strawberry plants. These results can support subsequent studies aiming to elucidate the action mode of biocontrol agents.

Philosophiae Doctor - PhD / We hypothesize that South African medicinal plants contain compounds that can act in synergism with synthetic antifungal compounds. Four fungicides - Sporekill™, Rovral™, Terminator™ and Teldor™ at doses 0.1, 0.2, 0.4 and 0.8 mL L-1 and plant species Galenia africana, Elytropappus rhinocerotis and Tulbaghia violacea were tested aloneand in different combinations for their potency (efficacy) on radial growth inhibition of Botrytis cinerea strains on potato dextrose plates. Four doses of plant extract for each of the respective plant species were used. A total of 48 combinations were tested for each strain. Mixtures of plant extracts were far more effective in controlling strains compared to the individual components alone, representing significant levels of in vitro synergistic interactions. Combinations of these components represent an attractive future prospect for the development of new management strategies for controlling B. cinerea. Since the in vitro tests of these mixtures showed inhibitory activity, the mixtures were tested for activity in assays on Granny Smith apples. In vitro tests can be used to screen mixtures to obtain information on their inhibitory activity on a pathogen, however, the environmental conditions of the fruit and the ability of the pathogen to grow into the fruit cannot be simulated in vivo. A series of two-fold doses of medicinal plant extracts were combined with fungicides to conduct decay inhibition studies. The incidence of gray mold was significantly reduced by mixtures of plant extracts and fungicides. Under conditions similar to those in commercial storage, a drench treatment with G. africana and Rovral™ significantly (p=0.05) inhibit gray mold on the apples and was more effective than the plant extract and fungicide alone. The treatments exerted synergistic effects and were markedly better than the components applied alone. The wound colonization assay was used for optimal decay control. In a drench, much higher volumes of the treatments are used to ensure that the components of the suspension are deposited evenly over the entire fruit surface. Drenching of fruit to apply other chemicals is an established practise in the pome (fleshy) fruit industry, and simplifies the commercial application of the mixtures, as no additional infrastructure at commercial packing houses will be required. This approach not only makes it possible to reduce fungicide concentrations while maintaining adequate decay control, but also ensures a reduction of the chemical residue on the fruit. / South Africa

Philosophiae Doctor - PhD / We hypothesize that South African medicinal plants contain compounds that can act in synergism with synthetic antifungal compounds. Four fungicides - Sporekill™, Rovral™, Terminator™ and Teldor™ at doses 0.1, 0.2, 0.4 and 0.8 mL L-1 and plant species Galenia africana, Elytropappus rhinocerotis and Tulbaghia violacea were tested alone and in different combinations for their potency (efficacy) on radial growth inhibition of Botrytis cinerea strains on potato dextrose plates. Four doses of plant extract for each of the respective plant species were used. A total of 48 combinations were tested for each strain. Mixtures of plant extracts were far more effective in controlling strains compared to the individual components alone, representing significant levels of in vitro synergistic interactions. Combinations of these components represent an attractive future prospect for the development of new management strategies for controlling B. cinerea. Since the in vitro tests of these mixtures showed inhibitory activity, the mixtures were tested for activity in assays on Granny Smith apples. In vitro tests can be used to screen mixtures to obtain information on their inhibitory activity on a pathogen, however, the environmental conditions of the fruit and the ability of the pathogen to grow into the fruit cannot be simulated in vivo. A series of two-fold doses of medicinal plant extracts were combined with fungicides to conduct decay inhibition studies. The incidence of gray mold was significantly reduced by mixtures of plant extracts and fungicides. Under conditions similar to those in commercial storage, a drench treatment with G. Africana and Rovral™ significantly (p=0.05) inhibit gray mold on the apples and was more effective than the plant extract and fungicide alone. The treatments exerted synergistic effects and were markedly better than the components applied alone. The wound colonization assay was used for optimal decay control. In a drench, much higher volumes of the treatments are used to ensure that the components of the suspension are deposited evenly over the entire fruit surface

The circadian clock regulates a wide range of biological processes, allowing plants to be prepared for predictable daily diurnal changes in environmental cues such as light and temperature. Recent studies have suggested that the circadian clock may also control plant immunity. The exact nature of the interaction between the circadian clock and plant pathogens remains unknown. Our focus in this study is on the elucidation of the role of the biological clock in plant immunity against the necrotrophic pathogen to Botrytis cinerea. In order to do this we tested the level of susceptibility to B. cinerea in Arabidopsis thaliana wild type and transgenic plants: toc1, cca1/lhy, cca1/toc1, lhy/toc1, cca1/lhy/toc1, GLK1 OE, GLK2 OE, glk1, glk2, and glk1/glk2. We demonstrated that the time of infection plays a role in susceptibility to B. cinerea. Specifically, we found that plants are more susceptible to infection in the subjective morning. We also found that genetic mutations in core clock components or in GLK genes leads to changes in susceptibility to B. cinerea. Our data suggests that clock genes are not solely responsible for plant immune responses to B. cinerea but rather the ways in which the biological clock system regulates outcome pathways. Furthermore, when we entrain the biological clock by changing the photoperiod (day length) in normal earth conditions LD 24h and SD 24h, we observed that short day plants had higher susceptibility to B. cinerea than long day plants. In addition, when we entrain the biological clock in different photoperiods, the LD 30h photoperiod plants displayed similar responses as those in the SD 24h photoperiod. The data indicates that day length is not responsible for the control of plant immunity; it is the ability of light to entrain the biological clock that is important. Together, the data strongly support the conclusion that the circadian clock plays a role in plant defense regulation.

Orientador: Marta Cristina Teixeira Duarte / Dissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Engenharia de Alimentos / Made available in DSpace on 2018-08-27T13:08:18Z (GMT). No. of bitstreams: 1 Almeida_GustavoSteffende_M.pdf: 767493 bytes, checksum: c194394bcc80ba1852de47976dba16ad (MD5) Previous issue date: 2015 / Resumo: A cultura do morango é de grande importância no contexto agrícola mundial, com uma produção anual que supera 4,5 milhões de toneladas. Porém, as características desta fruta a tornam altamente susceptível a fatores físicos e biológicos de deterioração, tornando-a altamente perecível e acarretando altos níveis de perdas no campo e na pós-colheita. Dentre os fatores biológicos tem destaque a ação de fungos fitopatogênicos, dentre os quais o 'Botrytis cinerea' e o' Colletotrichum gloeosporioides' tem papel principal. Dentre as alternativas naturais para controle destes fitopatógenos, os óleos essenciais de plantas medicinais e aromáticas vêm sendo investigados. Neste contexto, no presente trabalho a atividade antifúngica de óleos essenciais de nove espécies de 'Citrus' e de 'Cymbopogon citratus' foi avaliada in vitro contra 'B. cinerea' e' C. gloeosporioides'. Os óleos de 'Citrus sinensis' e 'C. citratus' foram os mais ativos in vitro (MIC ¿ 0,25 e 0,06 mg.mL-1, respectivamente) e, portanto, selecionados para os ensaios com morangos in natura, visando manter sua qualidade pós-colheita. Os ensaios foram conduzidos sob duas condições ambientais, ou seja, sob condições ambiente de armazenamento (25 ºC) e em armazenamento refrigerado (10 ºC). Aplicados na concentração 1,0 mg.mL-1 , o óleo essencial de 'C. sinensis' mostrou-se mais eficaz sob condições ambiente de armazenamento, e a mistura dos dois óleos em partes iguais em armazenamento refrigerado. Em ambas as condições, melhores resultados foram obtidos através da aplicação por contato direto. Os óleos essenciais foram ainda incorporados a um filme de revestimento composto por matriz de metil-celulose e testados in vitro quanto ao seu potencial de inibição de 'B. cinerea'. Nestas condições, o óleo essencial de 'C. sinensis' mostrou-se o mais eficiente. Através dos resultados foi possível concluir que os óleos essenciais são uma opção viável para o controle de fitopatógenos em morango e para manutenção de sua qualidade pós-colheita / Abstract: The strawberry crop has great importance in agricultural world wide context, with an annual production exceeding 4.5 million tons. However, the characteristics of this fruit make it highly susceptible to physical and biological damage, making it easily perishable and causing losses in field and postharvest. Among the biological factors, the action of phytopathogenic fungi such as 'Botrytis cinerea' and "Colletotrichum gloeosporioides" have a main role. Natural alternatives have been proposed in order to replace synthetic products currently used in control of these plant pathogens, of which the essential oils from medicinal and aromatic plants have demonstrated good potential. In this context, the present study evaluated the in vitro antifungal activity of the essential oils from nine 'Citrus' species and from' Cymbopogon citratus' against 'B. cinerea' and 'C. gloeosporioides'. 'Citrus sinensi's and 'C. citratus' essential oils showed highest activities (MIC values 0.25 mg.mL-1 and 0.06 mg.mL-1, respectively) and were selected for the assays with in natura strawberries aiming at maintaining its postharvest quality. The tests were conducted under two simulated environmental conditions, ie, environment conditions of storage (25 ºC) and refrigerated storage (10 ºC). The crude essential oil of 'C. sinensis' (1.0 mg.mL-1) was the more effective in environmental conditions of storage, and the mixture of the two oils (1:1 ¿ 1.0 mg.mL-1) in refrigerated storage. In both conditions, best results were obtained when the oils were applied by direct contact. The essential oils were also incorporated into a coating film consisting of methyl cellulose matrix and tested to their 'in vitro' inhibitory potential against 'B. cinerea'. Accordingly, the essential oil of 'C. sinensis' proved to be the most effective as antifungal. The results allowed conclude that essential oils are a viable option for controlling phytopathogenic fungi in strawberry and for continuing postharvest quality / Mestrado / Ciência de Alimentos / Mestre em Ciência de Alimentos