Virginia ranks third in fresh market staked tomato (Lycopersicon esculentum Mill.) production with approximately 1,659 hectares on the Eastern Shore. Estimated annual gross value is $30,800,000. Copper and chlorothalonil have long been considered essential to control bacterial and fungal diseases in fresh market tomatoes. High rates of these fungicides on tomatoes grown under plastic mulch have led to concerns about their potential adverse effect on water quality in estuaries adjacent to fields. The development of new fungicides, such as azoxystrobin and acibenzolar-S-methyl, which have more favorable environmental fate characteristics and are used at much lower rates, may provide viable alternatives to copper and chlorothalonil. Using a disease forecasting system, such as Tomcast, may reduce the number of applications of fungicides. The research objectives of this study were to reduce the amount of copper and chlorothalonil used in fresh market tomato production and to evaluate the effectiveness of the Tomcast disease forecasting system for controlling fungal leaf diseases on the Eastern Shore. Field studies compared copper to acibenzolar-S- methyl for bacterial diseases caused by Xanthomonas campestris pv. vesicatoria and Pseudomonas syringae pv. tomato. Research plots were established in a randomized complete block design with four replications in grower fields and at the Eastern Shore Agricultural Research & Extension Center. Acibenzolar proved to be as effective as the standard copper bactericides in controlling bacterial spot. Acibenzolar provided better control than the standard copper bactericides when bacterial speck was the target disease. Azoxystrobin application alternated with maneb was evaluated as a replacement for chlorothalonil. Azoxystrobin and Tomcast were studied as tools to reduce chlorothalonil use for control of Alternaria solani. Tomcast can reduce the number of applications by 40-70 % per year and provide adequate control of early blight. Azoxystrobin provides better control of early blight than chlorothalonil. Use of these new, more environmentally compatible, plant-protection products , along with the Tomcast disease forecasting system, can significantly reduce or eliminate the need to use copper and chlorothalonil for tomato disease management and therefore eliminate them as potential pollutants of the Chesapeake Bay and Atlantic Ocean estuaries. / Master of Science

In 2007 and 2008 more than 100 million dollars of fresh market tomatoes were grown in Virginia, with the majority of production occurring on the Eastern Shore of Virginia (ESV), according to the National Agricultural Statistics Service. Bacterial wilt of tomato, caused by Ralstonia solanacearum (Smith) and Yabucchi et al., is the most devastating disease of tomato on the ESV. Four 'observational trials' were conducted on the ESV over three growing seasons to determine the temporal and spatial distribution of this disease in commercial tomato fields. Plants were assessed at approximately one-week intervals throughout the growing seasons and the incidence of bacterial wilt for each individual plant was recorded. A steady increase in both disease incidence and clustered distribution of the disease within rows was observed as the growing season progressed. Positive correlations between disease incidence and percentage of rows exhibiting a significant clustered distribution occurred in all trials, which indicated an increase in clustered distribution as disease incidence increased. Research trials were conducted over three years, beginning in the summer of 2007, to investigate the effects of tomato bacterial wilt resistant cultivars on the ESV. In 2008 and 2009, the selective, systemic compound which induces host plant resistance, acibenzolar-S-methyl (ASM) was incorporated into resistant cultivar trials. Results from the 2007 trial revealed significant resistance in some of the breeding lines, CRA 66 and PI 126408. The 2008 and 2009 trials revealed that ASM was not effective at reducing levels of bacterial wilt. Grafted transplants in the spring trials of 2008 and 2009 had varied results in resistance and yield. Results revealed the tomato cultivar BHN 669 was an excellent resistant cultivar with promising yield potential and fruit quality. / Master of Science

Tobacco SABP2, a 29. kDa protein catalyzes the conversion of methyl salicylic acid (MeSA) into salicylic acid (SA) to induce SAR. Pretreatment of plants with acibenzolar-. S-methyl (ASM), a functional analog of salicylic acid induces systemic acquired resistance (SAR). Data presented in this paper suggest that SABP2 catalyzes the conversion of ASM into acibenzolar to induce SAR. Transgenic SABP2-silenced tobacco plants when treated with ASM, fail to express PR-1 proteins and do not induce robust SAR expression. When treated with acibenzolar, full SAR is induced in SABP2-silenced plants. These results show that functional SABP2 is required for ASM-mediated induction of resistance.

Espécies de Phytophthora tem se destacado ao longo da história devido ao seu potencial destrutivo, se iniciando com a devastadora P. infestans na Irlanda e se estende até os dias de hoje com P. nicotianae em citros e P. plurivora em faia. Uma característica importante deste grupo de patógenos é que as medidas de controle da doença se baseiam na prevenção da entrada do patógeno na área visto que, uma vez instalado, o produtor precisa conviver com o mesmo, pois não se dispõem de métodos efetivos de controle. Neste sentido, a busca por métodos de controle torna-se primordial. O endofítico radicular Piriformospora indica, tem-se destacado em vários patossistemas devido a sua habilidade de induzir resistência contra patógenos, aumentar a tolerância à estresses abióticos e promover o crescimento de plantas. Taxtomina A, produzida por Streptomyces scabies, é capaz de ativar mecanismos de defesa de plantas, os quais são efetivos contra agentes patogênicos. Objetivou-se com este trabalho avaliar o efeito de P. indica e da taxtomina A sobre P. nicotianae em citros e P. plurivora em faia. Ambos foram avaliados quanto ao seu efeito direto sobre os patógenos em questão. O indutor de defesa vegetal Bion® foi utilizado em alguns ensaios para fins de comparação. Plântulas de citros e faia foram tratadas com concentrações crescentes de taxtomina e parâmetros fisiológicos, bioquímicos e de controle da doença foram avaliados. Taxtomina A não apresenta efeito direto sobre os patógenos avaliados. Os dados de incidência da doença em plântulas de faia tratadas com taxtomina A nas concentrações de 10, 25, 50 e 100 μg se mostraram consistentes com a quantidade de DNA do patógeno no sistema radicular, demonstrando que, aparentemente, a toxina induziu suscetibilidade nas plântulas de faia. Em citros, para os parâmetros fisiológicos e bioquímicos avaliados, em linhas gerais, a taxtomina A nas concentrações de 50 e 100 μg demonstrou potencial de aplicação no patossistema citros - P. nicotianae. Quando avaliada a mortalidade de plantas inoculadas com o patógeno e tratadas com taxtomina, bem como, quando quantificado o DNA do oomiceto no sistema radicular, as referidas concentrações também apresentaram os melhores desempenhos. Plântulas das mesmas espécies foram submetidas a inoculação com P. indica, sendo avaliados os efeitos na promoção de crescimento, na atividade de enzimas e de genes relacionados ao processo de defesa, bem como, no controle da doença. Não foi observado efeito direto do endofítico radicular sobre os patógenos avaliados. Quando plântulas de citros foram inoculadas com P. indica e depois com P. nicotianae, não foi observada promoção de crescimento e controle da doença. As análises histológicas e moleculares demonstraram a presença do endofítico no sistema radicular de plântulas de citros e faia. Análises bioquímicas revelaram apenas aumentos pontuais no teor de proteínas e na atividade da β-1,3-glucanase e da peroxidase no tratamento com P. indica + P. nicotianae. Os genes PR-1.4, PR-1.8, PR-β-glucosidase e Hsp70 foram induzidos em plântulas inoculadas com P. indica e com o patógeno, bem como no tratamento com Bion® e patógeno, porém em menor magnitude. O endofítico P. indica ativa o sistema de defesa de plântulas de citros, no entanto, os mecanismos ativados não são efetivos para o controle da doença na interação citros - P. nicotianae. / Phytophthora species has been important throughout history because of its destructive potential, starting with the devastating P. infestans in potatos in Ireland and extending to the present day with Phytophthora nicotianae on citrus and Phytophthora plurivora on beech. An important feature of this group of pathogens is that the disease control measures are based upon prevention of pathogen entry into the area since, once installed, producers have to live with it, because they do not have effective methods of control. Accordingly, the search for control methods becomes essential. The root endophytic fungus Piriformospora indica has been shown to induce resistance against pathogens, increase tolerance to abiotic stresses and promote the growth of plants. On the other hand, the phytotoxin thaxtomin A, produced by Streptomyces scabies, is able to activate plant defense mechanisms, which are effective against pathogens. Thus, the objective of this study was to evaluate the effect of P. indica and thaxtomin A on the control of P. nicotianae on citrus and P. plurivora on beech. Both were assessed for their direct effect on these pathogens. The plant defense inducer Bion® was used in some tests for comparison. Besides that, seedlings of citrus or beech were treated with P. indica or increasing concentrations of thaxtomin and physiological / biochemical parameters as well as those related to induction of resistance and disease control were evaluated. The results showed that thaxtomin A did no exhibit any direct effect on the pathogens. The incidence of disease on beech seedlings treated with thaxtomin A, in concentrations of 10, 25, 50 and 100 ug, were consistent with the amount of DNA of the pathogen in the root tissue, demonstrating that, apparently, the toxin induced susceptibility in the seedlings. In citrus, for the physiological and biochemical parameters evaluated, in general, the thaxtomin in the concentrations of 50 and 100 ug showed potential for application in the citrus pathosystem - P. nicotianae. When the mortality of the seedlings treated with thaxtomin and inoculated with the pathogen as well as the oomycete DNA amount in the root system were evaluated these concentrations also had the best performances. As mentioned above, seedlings of beech or citrus were also inoculated with the endophyte and its effect on growth promotion, enzyme activities, the expression of genes related to defense process and on disease control was measured. There was no direct effect of the root endophyte on the pathogens. When citrus seedlings were inoculated with P. indica and then with P. nicotianae, there was no growth promotion or disease control. Histological and molecular analysis showed the presence of the endophytic fungus inside the root system of citrus and beech seedlings. Biochemical analyzes revealed only occasional increases in protein content and the activities of β-1,3-glucanase and peroxidase in the treatment with P. indica + P. nicotianae in citrus seedlings. The genes PR-1.4, PR- 1.8, PR-β-glucosidase and Hsp70 were induced in seedlings inoculated with P. indica and challenged with the pathogen as well as in the treatment with the resistance inducer Bion® plus pathogen, but to a lesser magnitude. Finally, the endophytic P. indica is able to active defense system in citrus, however, the activated mechanisms seems not to be effective in controlling the disease in the interaction citrus - P. nicotianae.

Submitted by (lucia.rodrigues@ufrpe.br) on 2017-02-17T13:39:56Z No. of bitstreams: 1 Camila Cristina Lage de Andrade.pdf: 845467 bytes, checksum: ef8d7e269cd9b79e14fc8dbcd17e8edd (MD5) / Made available in DSpace on 2017-02-17T13:39:56Z (GMT). No. of bitstreams: 1 Camila Cristina Lage de Andrade.pdf: 845467 bytes, checksum: ef8d7e269cd9b79e14fc8dbcd17e8edd (MD5) Previous issue date: 2012-02-16 / Conselho Nacional de Pesquisa e Desenvolvimento Científico e Tecnológico - CNPq / The bacterial speck caused by Pseudomonas syringae pv. tomato (Pst) (Okabe) Young, Dye & Wilkie has economic relevance to the tomato for industry in Brazil. The use of products that potentiate and/or induce plant defenses is an alternative that meets the requirements of integrated disease management. This study aimed to evaluate the effect of silicon (Si) and chemical inducers in some components of tomato resistance to bacterial speck and activity of enzymes involved in plant defense. In the first study, tomato plants were grown in: soil without calcium silicate (control) (T1), soil without calcium silicate and plants sprayed with Supa Silica® (SS) (2 mL/L SS) (T2) and soil with calcium silicate (0.38 g) (T3), being inoculated by spraying with a suspension of the pathogen. Our research evaluated the incubation period (IP), number of lesions (NL) per plant, severity estimated by the software QUANT 1.0 (SEQ) and leaf concentration of Si, as well as the activity of enzymes peroxidases (POX), polyphenoloxidases (PPO) β-1,3-glucanases (GLU), phenylalanine ammonia lyases (PAL), lipoxygenases (LOX) and concentration of malondialdehyde (MDA). It was assessed the effect of SS in the Pst growth in vitro. There was no significant difference between treatments for the IP and the foliar concentration of Si. No significant differences were found for the NL per plant and SEQ between T3 and control. T2 significantly reduced NL in 46.8 and 45.1% and SEQ in 61.5 and 56.2% when compared with control and T3, respectively. There was negative linear response of SS doses on the Pst growth in vitro. The activity of POX, PPO and GLU was significantly higher in T2 and T3. The activity of FAL and LOX was significantly higher in T3. The MDA concentration was significantly higher in T2 compared to control, in non-inoculated plants and at 7 days after inoculation (d.a.i.), being significantly lower at 10 d.a.i. In the second study, three experiments were conducted to evaluate the effect of jasmonic acid (JA; 0.1 mM), ethephon (ET, 0.5 mM) and acibenzolar-S-methyl (Bion®, ASM; 300 mg/L) sprayed 48 h before inoculation with Pst, evaluating the IP and NL per plant, and the activity of enzymes POX, PPO, GLU and LOX. It was also assessed the effect of JA and ET in the growth of Pst in vitro. Only in experiment 3 incubation period increased significantly by one day in the plants sprayed with ASM compared with control. In all experiments, NL per plant was significantly reduced by JA, ET e ASM in relation to the control reaching values of 38.9, 45.3 and 68.1%, respectively, in experiment 2. The growth of Pst in vitro was not significantly influenced. In some evaluation times JA has significantly raised the activity of POX, PPO and GLU; ASM has elevated the activity of PPO, GLU and LOX, while ET only significantly raised the activity of GLU and LOX compared to the control. The results of these studies suggest that spraying tomato plants with SS and the inducers JA, ET and ASM affected some components of tomato resistance to bacterial speck, and activated the enzymes POX, PPO, GLU, PAL and LOX, involved in defense responses of tomato plants to Pst. / A pinta bacteriana causada por Pseudomonas syringae pv. tomato (Pst) (Okabe) Young, Dye & Wilkie tem importância econômica para a cultura do tomateiro industrial no Brasil. O uso de agentes potencializadores e/ou indutores das defesas das plantas é uma alternativa que atende aos requisitos do manejo integrado de doenças. Este estudo avaliou o efeito do silício (Si) e de indutores químicos em alguns componentes de resistência do tomateiro à pinta bacteriana e na atividade de enzimas envolvidas na defesa das plantas. No primeiro estudo plantas de tomateiro foram cultivadas em: solo sem silicato de cálcio (controle) (T1), solo sem silicato de cálcio e plantas pulverizadas com Supa Sílica® (SS) (2 mL/L de SS) (T2) e solo com silicato de cálcio (0,38 g) (T3), sendo inoculadas por pulverização com suspensão do patógeno. Foram avaliados o período de incubação (PI), número de lesões (NL) por planta, severidade estimada pelo software QUANT 1.0 (SEQ) e concentração foliar de Si, bem como a atividade das enzimas peroxidases (POX), polifenoloxidases (PFO), β-1,3-glucanases (GLU), fenilalanina amônia-liases (FAL), lipoxigenases (LOX) e concentração de aldeído malônico (MDA). Também foi avaliado o efeito do SS no crescimento de Pst in vitro. Não houve diferença significativa entre os tratamentos para o PI e para a concentração foliar de Si. Não houve diferença significativa para o NL por planta e a SEQ entre o tratamento T3 e o controle. O tratamento T2 reduziu significativamente o NL 46,8 e 45,1% e a SEQ 61,5 e 56,2%, em relação ao controle e ao tratamento T3, respectivamente. Houve resposta linear negativa das doses de SS no crescimento da Pst in vitro. A atividade das POX, PFO e GLU foi significativamente maior nos tratamentos T2 e T3. A atividade das FAL e das LOX foi significativamente maior no T3. A concentração de MDA foi significativamente maior no tratamento T2 em relação ao controle, nas plantas não inoculadas com Pst e aos 7 dias após inoculação (d.a.i.); sendo significativamente menor aos 10 d.a.i. No segundo estudo, três experimentos foram realizados para avaliar o efeito dos indutores ácido jasmônico (AJ; 0,1 mM), ethephon (ET; 0,5 mM) e acibenzolar-S-metil (Bion®, ASM; 300 mg/L) pulverizados 48 h antes da inoculação da Pst, avaliando-se o PI e o NL por planta, além da atividade das enzimas POX, PFO, GLU e LOX. Também foi avaliado o efeito do AJ e do ET no crescimento da Pst in vitro. Apenas no experimento 3 o PI aumentou significativamente em 1 dia nas plantas pulverizadas com ASM em relação ao controle. O NL por planta foi significativamente reduzido pelos tratamentos AJ, ET e ASM em relação ao controle para todos os experimentos, atingindo valores de 38,9, 45,3 e 68,1%, respectivamente, no experimento 2. O crescimento da Pst in vitro não foi influenciado significativamente pelo AJ ou ET. Em determinadas épocas de avaliação, o AJ elevou significativamente a atividade das POX, PFO e GLU; ASM elevou a atividade das PFO, GLU e LOX e o ET elevou a atividade das GLU e LOX em relação ao controle. Os resultados desses estudos evidenciam que a pulverização com SS e com indutores AJ, ET e ASM afetaram alguns dos componentes de resistência do tomateiro à pinta bacteriana, além de potencializar as enzimas POX, PFO, GLU, FAL e LOX, relacionadas com a defesa das plantas em resposta à Pst.

CoordenaÃÃo de AperfeiÃoamento de NÃvel Superior / O melÃo tem grande importÃncia para a economia brasileira, sendo sua produÃÃo exportada principalmente para os paÃses da UniÃo EuropÃia. Assim, à fundamental o controle de doenÃas pÃs-colheita deste fruto. O Fusarium pallidoroseum à um importante fitopatÃgeno que provoca podridÃes no melÃo. Esta doenÃa representa um obstÃculo sÃrio em sua comercializaÃÃo. Este trabalho avaliou os efeitos do BTH, um anÃlogo estrutural e funcional do Ãcido salicÃlico, nas respostas bioquÃmicas da defesa do melÃo desafiado com o F. pallidoroseum e do meloeiro nÃo desafiado. Doze horas depois de colhidos, os melÃes foram mergulhados em soluÃÃes de BTH (concentraÃÃes de 0,5, 1,0, e 2,0 mM de ingrediente ativo) e, depois 60 horas, foram inoculados com o fungo. Amostras dos frutos (2 cm diÃmetro x 1 cm de profundidade), prÃximos ao local de infecÃÃo, foram retiradas em 3, 7 e 10 dias apÃs inoculaÃÃo, pesadas e armazenadas à -84 ÂC atà serem utilizadas. Plantas do melÃo de 8 dias foram borrifadas com 300 ÂL de BTH nas concentraÃÃes de 0,3, 0,5, e 1,0 mM. As folhas secundÃrias foram colhidas em 2, 4, 6, 8, 10, 12, e 14 dias apÃs tratamento. AlÃm da avaliaÃÃo do sintoma nos frutos, extratos totais do fruto e das folhas secundÃrias foram preparados com tampÃo acetato 50 mM, pH 5.2, contendo 150 mM de NaCl, e os Ãndices de proteÃna e as atividades enzimÃticas da peroxidase (POX), da fenilalanina amÃnia liase (PAL), da β-1,3-glucanase (GLU), da peroxidase do ascorbato (APX) e da superoxide dismutase (SOD) foram medidos. Observou-se que BTH nÃo reduziu significativamente a incidÃncia e a severidade da podridÃo causada pelo patÃgeno. Nem 2 mM de BTH modificou significativamente as atividades de enzimas relacionadas a defesa do fruto. Ao contrÃrio, nas plantas do melÃo, BTH aumentou as atividades da POX, GLU e da SOD, mas nÃo modificou a PAL e APX foi inibida. Estes resultados sugerem que BTH nÃo trabalhou como um indutor de defesas bioquÃmicas no melÃo, mas induziu respostas de defesa nas plantas. ConseqÃentemente, sugere-se que BTH poderia ser usado como uma estratÃgia tecnolÃgica para a proteÃÃo de frutas do melÃo contra a podridÃo causada pelo F. pallidoroseum atravÃs da induÃÃo das respostas bioquÃmicas de defesa da prÃpria planta, que, provavelmente, estarà transferindo estas caracterÃsticas aos frutos. Entretanto, esta hipÃtese que està sendo proposta necessita ser avaliada. / Melon fruit constitutes one of the main segments of the Brazilian economy. Its production is exported particularly to countries in the European Union. Thus it is fundamental the control of postharvest diseases of melon. Fusarium pallidoroseum is an important phytopathogen which provokes rot in melon fruits. This disease represents a serious obstacle in its commercialization as a foreign commodity. In this present work the effects of BTH, a structural and functional analogue of salicylic acid, on the biochemical defense responses of melon fruits challenged with F. pallidoroseum and of unchallenged melon plants were assessed. Twelve hours after harvesting melon fruits were immersed in BTH (0.5, 1.0, and 2.0 mM concentrations of active ingredient) and 60 hours later inoculated with the fungus. Fruit cuts (2 cm diameter x 1 cm deep), close to the inoculation sites, were excised at 3, 7, and 10 days after fungal inoculation, weighed and kept at -84 ○C until used. Eight day-old melon plants were sprayed with 300 ÂL BTH at 0.3, 0.5, and 1.0 mM concentrations. Secondary leaves were harvested at 2, 4, 6, 8, 10, 12, and 14 hours after sprayings. Besides to symptom evaluation in melon fruits, crude extracts from the fruit cuts and leaves were prepared with 50 mM acetate buffer, pH 5.2, containing 150 mM NaCl, and the protein contents and enzymatic activities of peroxidase (POX), phenylalanine ammonia lyase (PAL), β-1,3-glucanase (GLU), ascorbate peroxidase (APX), and superoxide dismutase (SOD) were measured. It was observed that BTH did not reduce significantly the incidence and severity of the rot caused by the pathogen. Neither 2 mM BTH significantly modify the activities of defense-related enzymes in melon fruits. Contrary, in the melon plants, BTH increased the activities of POX, GLU, and SOD, but did not modify PAL and further inhibited APX. These results suggest that BTH did not work as an inductor of biochemical defenses in melon fruits, but it induced defense responses in the melon plants. Therefore it is suggested that BTH could be used as a technological strategy for protection of melon fruits from the rot caused by F. pallidoroseum by means of induction of biochemical defense responses of the melon plant itself which will likely be transferring these traits to its fruits. However, this hypothesis that is being proposed needs to be assessed.

Espécies de Phytophthora tem se destacado ao longo da história devido ao seu potencial destrutivo, se iniciando com a devastadora P. infestans na Irlanda e se estende até os dias de hoje com P. nicotianae em citros e P. plurivora em faia. Uma característica importante deste grupo de patógenos é que as medidas de controle da doença se baseiam na prevenção da entrada do patógeno na área visto que, uma vez instalado, o produtor precisa conviver com o mesmo, pois não se dispõem de métodos efetivos de controle. Neste sentido, a busca por métodos de controle torna-se primordial. O endofítico radicular Piriformospora indica, tem-se destacado em vários patossistemas devido a sua habilidade de induzir resistência contra patógenos, aumentar a tolerância à estresses abióticos e promover o crescimento de plantas. Taxtomina A, produzida por Streptomyces scabies, é capaz de ativar mecanismos de defesa de plantas, os quais são efetivos contra agentes patogênicos. Objetivou-se com este trabalho avaliar o efeito de P. indica e da taxtomina A sobre P. nicotianae em citros e P. plurivora em faia. Ambos foram avaliados quanto ao seu efeito direto sobre os patógenos em questão. O indutor de defesa vegetal Bion® foi utilizado em alguns ensaios para fins de comparação. Plântulas de citros e faia foram tratadas com concentrações crescentes de taxtomina e parâmetros fisiológicos, bioquímicos e de controle da doença foram avaliados. Taxtomina A não apresenta efeito direto sobre os patógenos avaliados. Os dados de incidência da doença em plântulas de faia tratadas com taxtomina A nas concentrações de 10, 25, 50 e 100 μg se mostraram consistentes com a quantidade de DNA do patógeno no sistema radicular, demonstrando que, aparentemente, a toxina induziu suscetibilidade nas plântulas de faia. Em citros, para os parâmetros fisiológicos e bioquímicos avaliados, em linhas gerais, a taxtomina A nas concentrações de 50 e 100 μg demonstrou potencial de aplicação no patossistema citros - P. nicotianae. Quando avaliada a mortalidade de plantas inoculadas com o patógeno e tratadas com taxtomina, bem como, quando quantificado o DNA do oomiceto no sistema radicular, as referidas concentrações também apresentaram os melhores desempenhos. Plântulas das mesmas espécies foram submetidas a inoculação com P. indica, sendo avaliados os efeitos na promoção de crescimento, na atividade de enzimas e de genes relacionados ao processo de defesa, bem como, no controle da doença. Não foi observado efeito direto do endofítico radicular sobre os patógenos avaliados. Quando plântulas de citros foram inoculadas com P. indica e depois com P. nicotianae, não foi observada promoção de crescimento e controle da doença. As análises histológicas e moleculares demonstraram a presença do endofítico no sistema radicular de plântulas de citros e faia. Análises bioquímicas revelaram apenas aumentos pontuais no teor de proteínas e na atividade da β-1,3-glucanase e da peroxidase no tratamento com P. indica + P. nicotianae. Os genes PR-1.4, PR-1.8, PR-β-glucosidase e Hsp70 foram induzidos em plântulas inoculadas com P. indica e com o patógeno, bem como no tratamento com Bion® e patógeno, porém em menor magnitude. O endofítico P. indica ativa o sistema de defesa de plântulas de citros, no entanto, os mecanismos ativados não são efetivos para o controle da doença na interação citros - P. nicotianae. / Phytophthora species has been important throughout history because of its destructive potential, starting with the devastating P. infestans in potatos in Ireland and extending to the present day with Phytophthora nicotianae on citrus and Phytophthora plurivora on beech. An important feature of this group of pathogens is that the disease control measures are based upon prevention of pathogen entry into the area since, once installed, producers have to live with it, because they do not have effective methods of control. Accordingly, the search for control methods becomes essential. The root endophytic fungus Piriformospora indica has been shown to induce resistance against pathogens, increase tolerance to abiotic stresses and promote the growth of plants. On the other hand, the phytotoxin thaxtomin A, produced by Streptomyces scabies, is able to activate plant defense mechanisms, which are effective against pathogens. Thus, the objective of this study was to evaluate the effect of P. indica and thaxtomin A on the control of P. nicotianae on citrus and P. plurivora on beech. Both were assessed for their direct effect on these pathogens. The plant defense inducer Bion® was used in some tests for comparison. Besides that, seedlings of citrus or beech were treated with P. indica or increasing concentrations of thaxtomin and physiological / biochemical parameters as well as those related to induction of resistance and disease control were evaluated. The results showed that thaxtomin A did no exhibit any direct effect on the pathogens. The incidence of disease on beech seedlings treated with thaxtomin A, in concentrations of 10, 25, 50 and 100 ug, were consistent with the amount of DNA of the pathogen in the root tissue, demonstrating that, apparently, the toxin induced susceptibility in the seedlings. In citrus, for the physiological and biochemical parameters evaluated, in general, the thaxtomin in the concentrations of 50 and 100 ug showed potential for application in the citrus pathosystem - P. nicotianae. When the mortality of the seedlings treated with thaxtomin and inoculated with the pathogen as well as the oomycete DNA amount in the root system were evaluated these concentrations also had the best performances. As mentioned above, seedlings of beech or citrus were also inoculated with the endophyte and its effect on growth promotion, enzyme activities, the expression of genes related to defense process and on disease control was measured. There was no direct effect of the root endophyte on the pathogens. When citrus seedlings were inoculated with P. indica and then with P. nicotianae, there was no growth promotion or disease control. Histological and molecular analysis showed the presence of the endophytic fungus inside the root system of citrus and beech seedlings. Biochemical analyzes revealed only occasional increases in protein content and the activities of β-1,3-glucanase and peroxidase in the treatment with P. indica + P. nicotianae in citrus seedlings. The genes PR-1.4, PR- 1.8, PR-β-glucosidase and Hsp70 were induced in seedlings inoculated with P. indica and challenged with the pathogen as well as in the treatment with the resistance inducer Bion® plus pathogen, but to a lesser magnitude. Finally, the endophytic P. indica is able to active defense system in citrus, however, the activated mechanisms seems not to be effective in controlling the disease in the interaction citrus - P. nicotianae.

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / The objectives of this work were divided in two experiments phases. In the first phase, it was studied the presence of trans-resveratrol in temperate climate fruits and the variation of their concentration in these fruits during storage period, as well the amount of total phenols and total anthocyanins. It was evaluated total phenols, total anthocyanins and trans-resveratrol, before and after cold storage and under controlled atmosphere; and after five days shelf-life at 20°C in Bruno kiwifruit; Araza and Yvahe strawberries; Black mulberry; Galaxi , Gala and Fuji apples; persimmon Fuyu ; Bleugem blueberry, Isabel , Merlot and Niagara Pink grapes. It was detected trans-resveratrol in natura in flesh Bruno kiwifruit; Araza and Yvahe strawberries; Black mulberry. In other fruits, it was detected transresveratrol in natura in the skin. The cold storage stimulated trans-resveratrol synthesis in Bleugem blueberry, and Isabel and Merlot grapes. In the second phase, it was evaluated the induction of trans-resveratrol synthesis and its possible action as phytoalexin in Gala and Fuji apples stored in controlled atmosphere were evaluated. Experiments were with the application of abiotic elicitors: ultraviolet irradiation, phosphite and acibenzolar-S-methyl, applied before controlled atmosphere storage, and intermittent ozone applied during storage of to Gala and Fuji apples. Rot was controlled in another experiment with the same elicitors. In fruit bark it was analyzed trans-resveratrol; total phenols and total anthocyanins, as well as the diameter of injury after inoculation of Penicillium sp. The abiotic elicitors were induced in 'Fuji' apples trans-resveratrol synthesis in the following sequence: Acibenzolar-Smethyl > phosphite ³ UV-C irradiation ³ ozone, but it wasn t effective on 'Gala' apple. There was no correlation between the syntheses of trans-resveratrol and rotting control, with exception of phosphite was control of rot in 'Gala'. / Os objetivos deste trabalho foram alcançados em duas etapas. Na primeira etapa, o objetivo foi avaliar a presença de trans-resveratrol em frutos de clima temperado e a sua variação de concentração nesses frutos durante o período de armazenamento, assim como a evolução dos polifenóis totais e antocianinas totais. Avaliou-se o conteúdo de polifenóis totais, antocianinas totais e trans-resveratrol antes e após o armazenamento refrigerado e em atmosfera controlada, e após cinco dias de exposição a 20°C em kiwi Bruno , morangos Araza e Yvahe , amora (Morus nigra), maçãs Galaxi , Gala e Fuji , caqui Fuyu , mirtilo Bleugem , uvas Isabel , Merlot e Niágara Rosada . Detectou-se o trans-resveratrol in natura em polpa de kiwi Bruno , morangos Araza e Yvahe e amora Preta . Nos demais fruto, detectou-se in natura trans-resveratrol na casca. O armazenamento refrigerado estimulou a síntese de trans-resveratrol em mirtilo Bleugem , uvas Isabel e Merlot . Na segunda etapa, o objetivo foi avaliar a indução da síntese de trans-resveratrol e sua possível ação como fitoalexina em maçãs Gala e Fuji , armazenadas em atmosfera controlada. Realizaram-se experimentos com aplicação de elicitores abióticos: irradiação ultravioleta, fosfito e acibenzolar-S-metil, aplicados antes do armazenamento em atmosfera controlada, e ozônio aplicado de forma intermitente durante o armazenamento de maçãs Gala e Fuji . Realizou-se o controle de podridão em outro experimento com os mesmos elicitores. Na casca dos frutos analisaram-se trans-resveratrol, polifenóis totais e antocianinas totais, e o diâmetro de lesão após inoculação por Penicillium sp. Os elicitores abióticos induziram a síntese de trans-resveratrol em maçã Fuji na seguinte ordem: Acibenzolar-S-metil > Fosfito ³ irradiação UV-C ³ ozônio, mas não apresentaram efeito em maçãs Gala , durante o armazenamento em atmosfera controlada. Os elicitores abióticos não apresentaram efeito no controle de podridão, com exceção do fosfito que controlaram a podridão em maçã Gala . Não houve correlação entre a síntese de trans-resveratrol e o controle de podridão.

Low temperature injury is among the greatest challenges facing golf courses with ultradwarf bermudagrass (UDB) (Cynodon dactylon (L.) Pers. x C. transvaalensis Burtt-Davy) putting greens in Virginia. This research focused on the impact of turf covers, fungicide programming, core aeration, and trinexapac-ethyl (TE) on UDB cold tolerance, winter quality, and cold de-acclimation (CD). Our results indicate that the use of turf covers significantly increased UDB canopy and soil temperatures when air temperatures were below -3.9°C. Air gaps under covers and the use of double turf covers increased soil and canopy temperatures compared to single covers alone in some instances, but results were inconsistent. Late fall and early winter fungicide applications of chlorothalonil and azoxystrobin improved UDB quality throughout winter dormancy and spring green up. The addition of a pigmented phosphonate significantly improved winter and spring UDB quality. The addition of acibenzolar-S-methyl to fungicide programs did not improve winter UDB quality or spring green up. Summer core aeration programs were evaluated for their impact on spring green up, turfgrass quality, surface firmness, and moisture retention. Spring UDB green up was improved incrementally as surface disruption increased. Treatments with 20%, 15%, and 10% surface disruption produced higher color vs treatments with lower surface disruption. Surface firmness and volumetric water content of UDB were impacted by construction method but were not significantly impacted by core aeration programs. Field research revealed that 'fall only' and 'fall and winter' TE applications improved UDB quality but only 'fall and winter' delayed UDB premature CD in early spring when UDB can be susceptible to low temperature injury. Growth chamber studies evaluated the impact of TE on UDB cold tolerance to -9.4°C x time duration. Regression analysis predicted a 50% mortality exposure point for UDB under TE treatments of 9.84 hours at -9.4°C (r2=0.836) compared to 11.38 hours at -9.4°C (r2=0.671) for non-treated UDB during cold acclimation. Winter and spring scenarios resulted in delayed CD under TE but no differences in cold tolerance when exposed to -9.4°C. Together, these results increase our understanding of the impact of management practices on UDB winter quality, CD, and low temperature injury. / Doctor of Philosophy / Ultradwarf bermudagrass putting greens are commonly found on golf courses in warm climates. These grasses thrive in heat and humidity but are susceptible to injury or death when exposed to cold temperatures. This research is focused on evaluating management practices that may impact bermudagrass' susceptibility to injury from cold temperature exposure. The cultural practices evaluated include turf covers, fungicide programming, core aeration, and the use of plant growth regulators to manipulate the turfgrasses own self defense mechanisms. Our results show that the use of turf covers significantly increased putting green canopy and soil temperatures when air temperatures were below -3.9°C. Air gaps under covers and the use of double turf covers increased soil and canopy temperatures compared to single covers alone in some instances, but results were inconsistent. Late fall and early winter fungicide applications of commonly-used fungicides improved putting green quality throughout winter dormancy and spring green up. The addition of a green-pigmented phosphonate fungicide significantly improved winter and spring putting green quality. The addition of a plant defense activator, acibenzolar-S-methyl to fungicide programs did not improve winter quality or spring green up. Summer core aeration programs were evaluated for their impact on spring green up, turfgrass quality, surface firmness, and moisture retention. Spring green up was improved incrementally as surface disruption increased. Treatments with 20%, 15%, and 10% surface disruption produced higher color vs treatments with lower surface disruption. Surface firmness and soil moisture content of the putting greens were impacted by construction method but were not significantly impacted by core aeration programs. Field research revealed that 'fall only' and 'fall and winter' plant growth regulator applications improved ultradwarf bermudagrass quality but only 'fall and winter' delayed premature green-up in early spring when the turfgrass can be susceptible to low temperature injury. Growth chamber studies revealed that plants treated with the growth regulator, trinexapac-ethyl were more sensitive to low-temperature exposure than non-treated plants. Together, these results increase our understanding of the impact of management practices on UDB winter quality, CD, and low temperature injury.

Submitted by Mario BC (mario@bc.ufrpe.br) on 2017-03-16T14:18:00Z No. of bitstreams: 1 Ariele Carneiro de Andrade.pdf: 1066650 bytes, checksum: 26d95d8f20aec5260de9adb89d2195cd (MD5) / Made available in DSpace on 2017-03-16T14:18:00Z (GMT). No. of bitstreams: 1 Ariele Carneiro de Andrade.pdf: 1066650 bytes, checksum: 26d95d8f20aec5260de9adb89d2195cd (MD5) Previous issue date: 2015-07-15 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPES / The bell pepper (Capsicum annum L.) crop is one of the most important vegetables grown in Brazil, and has been showing significant growth. However, this culture can be affected by a number of diseases that cause economic damage and anthracnose, still represent a serious obstacle to production, both in the field and at home-de-vegetation. Anthracnose in Capsicum annum can be caused by several species of fungi of the genus Colletotrichum, but in Brazil, the majority of reports identify Colletotrichum gloeosporioides as the main causative agent of the disease. Infections characterized in lesions aerial parts of the plants such fruits, stems and leaves. The control method of the infections is used in fungicidal applications, however currently search for more sustainable alternatives. Thus the use of resistance inducers has been outstanding and can be applied preventively, by acting on the physiological and biochemical processes of plants. Given the above, the objective of this study was to investigate the efficacy of acibenzolar-S-methyl inducer for the control of anthracnose in pepper plants as well as the activation of enzymes related to pathogenicity and they peroxidase, polifeniloxidase, catalase, ascorbate peroxidase and β-1,3-glucanase.. The study was conducted in a greenhouse in Garanhuns-PE municipality, and the treatments were arranged in subdivided portion, in which the portion consisted of collection times and the subplot of different inducer of doses, with four replications. The plants were treated with the treatments as follows: 0.15g L-1; 0.30g L-1; 0.45g L-1, 0.60g L-1 acibenzolar-S-methyl (Syngenta®) and control (distilled water only). The results were analyzed by the software program SAEG 5.0 to 5% probability. The peroxidase activity was not significant for analysis of variance at 5% probability, however the dose 0.45g L-1 promoted greater activity. For peroxidase although not significant ascorbate activity to analysis of variance, the dose 0.15g L-1 showed increased activity of this enzyme is superior to the control. The increase in catalase activity was significant when the plants were treated with the highest dose of 0.60g L-1 inducer. On the eighth day after inoculation was higher by polyphenol oxidase activity, standing out from the others collection times. The dose 0.60g L -1 promoted greater activity of β-1,3-glucanase enzyme. When the plants medium subjected to the application of acibenzolar-S-methyl there was a reduction in the severity as the doses increased, thereby showing that prior treatment with resistance inducer consists of an efficient method for control of anthracnose disease. / A cultura do pimentão (Capsicum annum L.) é uma das mais importantes hortaliças cultivadas no Brasil. Entretanto essa cultura pode ser acometida por uma série de doenças que acarretam danos econômicos e a antracnose, ainda representa um sério entrave à produção, tanto em campo quanto em casa-de-vegetação. A antracnose nesta olerícola pode ser causada por várias espécies de fungos do gênero Colletotrichum, mas no Brasil, a maioria dos relatos identificam Colletotrichum gloeosporioides como o principal agente causal da doença. As infecções se caracterizam por lesões nas partes aéreas das plantas como nos frutos, caules e folhas. O método de controle das infecções mais utilizado consiste em aplicações de fungicidas, no entanto atualmente se busca por alternativas mais sustentáveis. Desse modo o uso de indutores de resistência vem se destacando podendo ser aplicado de forma preventiva, por atuar nos processos fisiológicos e bioquímicos das plantas. Diante do exposto, o objetivo deste estudo foi verificar a eficácia do indutor acibenzolar-S-metil para o controle da antracnose nas plantas de pimentão bem como a ativação de enzimas relacionadas à patogenicidade sendo elas peroxidase, polifeniloxidase, catalase, ascorbato peroxidase e β-glucanase. O estudo foi conduzido em casa de vegetação no município de Garanhuns-PE. Os tratamentos foram dispostos em parcela subdivida, na qual a parcela consistiu das épocas de coleta e a subparcela das diferentes doses do indutor, com quatros repetições. As plantas receberam os tratamentos da seguinte forma: 0,15g L-1, 0,30g L-1, 0,45g L-1, 0,60g L-1 do acibenzolar-S-metil (Syngenta®) e Controle (somente água destilada). Os resultados obtidos foram analisados pelo emprego do programa software SAEG 5.0 a 5% de probabilidade. A atividade peroxidase não foi significativa para análise de variância a 5% de probabilidade, no entanto a dose 0,45g L-1 promoveu maior atividade. Para a ascorbato peroxidase embora não significativo para análise de variância, a dose 0,15g L-1 apresentou maior atividade desta enzima sendo superior ao controle. O aumento na atividade catalase foi significativo quando as plantas foram tratadas com a maior dose do indutor 0,60g L-1. No oitavo dia após a inoculação houve maior atividade da enzima polifenoloxidase, destacando-se das demais épocas de coletas. A dose 0,60g L-1 promoveu maior atividade da β-1,3-glucanase. Quando as plantas foram submetidas à aplicação do acibenzolar-S-metil verificou-se uma redução da severidade à medida que as doses aumentavam, mostrando com isso que o tratamento prévio com indutor de resistência consiste em um método eficiente para o controle da doença antracnose.