L'utilisation de molécules stimulatrices de défense des plantes (SDP), également appelées inducteurs de résistance, constitute une alternative possible aux traitements fongicides conventionnels pour contrôler les maladies dues aux chanpignons phytopathogènes. Dans cette étude, nous avons mis en évidence que trois produits à caractère SDP (FSOV2, FSOV7 et FSOV10) protègent le blé (Triticum aestivum L.) contre la septoriose (Mycosphaerella graminicola, anamorphe Zymoseptoria tritici) lorsqu'ils sont utilisés de façon préventive, mais cette protection dépend fortement du cultivar considéré. Les cultivars Alixan, Premio et Altigo testés ici présentaient au départ des niveaux de résistance distincts à la septoriose. Les protections obtenues ne sont pas liées à un effet direct sur la germination des spores du champignon, mais à l'induction des mécanismes de défense chez le blé qui réduisent la nécrose foliaire et la sporulation du champignon. Ainsi, l'observation des différents stades du processus infectieux de M. graminicola en microscopie et le dosage des activités enzymatiques fongiques de dégradation des parois (CWDE) in planta révèlent que le niveau de protection induite varie avec le SDP appliqué et avec le cultivar traité. L'expression de neuf gènes impliqués dans différentes voies de défense, suivi par RTqPCR, et les activités enzymatiques peroxydase et phénylalanine ammonia lyase ont été mesurée au cours du temps, depuis le traitement par les SDP jusqu'à 5 jours après infection. Les résultats obtenus montrent, que les mécanismes de défense sont induits différemment en fonction du cultivar et en fonction du SDP appliqué. Ces résultats suggèrent que la réussite au champ des SDP est conditionnée de façon déterminante par le choix du couple SDP-cultivar. / The use of resistance inducers (RI) is a potential alternative to conventional fungicide treatments against plant fungal diseases. In the present study, we revealed that preventive applications of three RI conferred protection efficacies against M. graminicola, with protection levels varying with the wheat cultivar. Alixan, Premio and Altigo cultivars were previously known to exhibit distinct resistance levels to M. graminicola. The observed protections did not result from a direct effect on spore germination, but were related to the induction of wheat defense mechanisms. The induced resistances reduced foliar necrosis, as well as the sporulation level of the fungus. Microscopic observations of the infection process of M. graminicola and cell wall degrading enzymes (CWDE) activities measured in planta showed that the applied RI as well as the considered treated wheat cultivar influences the impact on the infection process and the protection efficacy. We investigated from the time of treatment until 5 days after inoculation plant peroxidase and phenylalanine ammonia lyase activities and the expression of nine genes involved in distinct defense pathways. Our results indicated that defense mechanisms are differently induced according both to the wheat cultivar and the RI. Therefore, the successful use of RI at the field level strongly depends on the RI-cultivar combination.

Bacillus subtilis Stämme vom FZB - FZB24, FZB37 und FZB38 des FZB Biotechnik, Berlin - und ihre Stoffwechselprodukte wurden als Agenzien für eine Resistenzinduktion gegen Schädiger in drei Wirtspflanzen-Erreger-Systemen, Vicia faba - Uromyces appendiculatus, Vicia faba - Aphis fabae und Triticum aestivum - Rhopalosiphum padi untersucht. Prä-inokulativ wurden Blätter und Saatgut der Pflanzen mit den Testsubstanzen behandelt. Zur Testung gelangten Bakterien-Kulturfiltrat (KF), KF-Zenrifugationsüberstände, die vegetativen Zellen und Sporen der B. subtilis Stämme. Nach lokaler Applikation der KF-Überstände von B. subtilis auf die Wirtspflanzenblätter konnte eine erhebliche Breite antibiotischer (entwicklungshemmender) Wirkungen auf A. fabae und R. padi beobachtet werden. Eine Untersuchung der freien Aminosäuren im Phloemsaft von Vicia faba zeigte, daß sich nach Befall von Aphis fabae speziell bei den zuvor mit bakteriellem Kulturfiltrat und KF-Überstand behandelten Pflanzen im Vergleich zu der nur mit Wasser behandelten Kontrolle, die Konzentration von neun Aminosäuren änderte. / In topical and systemic plant treatment, in three host–parasite systems, i.e. Vicia faba - Uromyces appendiculatus, Vicia faba - Aphis fabae and Triticum aestivum - Rhopalosiphum padi the culture filtrate and supernatant of Bacillus subtilis (FZB24, FZB37 and FZB38 from FZB Biotechnik Berlin) was shown to inhibit the development of urediospores produced by Uromyces appendiculatus. The performance of Aphis fabae and Rhopalosiphum padi was evaluated using life table tests where the aphids'' development time (tD), pre-reproduction time (td), relative growth rate (RGR) and intrinsic rate of natural increase (rm) were assessed. A wide range of antibiosis effects in Aphis fabae and Rhopalosiphum padi was observed when the supernatant of Bacillus subtilis was used as foliar topical treatment. The tested aphids presented longer development and pre-reproduction time; conversely a lower relative growth rate and intrinsic rate of natural increase was observed. The investigation of the free amino acids of the phloem sap of V. faba plants, showed the concentration of the amino acid serine remained unchanged in the supernatant induced plants in this study, which was interpreted as the major reason for the observed antibiosis effect on the tested aphids.

Os cogumelos Lentinula edodes (shiitake) e Agaricus blazei (cogumelo-do-sol) apresentam substâncias no corpo de frutificação (basidiocarpo) e no micélio com atividades antibióticas e imuno-regulatórias, havendo uma série de relatos sobre a atuação das mesmas no controle de doenças em animais. Em vegetais, não há informações sobre o efeito protetor do cogumelo-do-sol contra fitopatógenos. No caso de shiitake, embora pouco numerosos, os estudos mostraram o potencial do cogumelo para o controle de doenças de plantas, tais como a murcha bacteriana do tomateiro, a murcha de feijão-lima, além de doenças fúngicas em sorgo e da bacteriose do maracujazeiro. Os objetivos do presente trabalho foram o de avaliar o efeito de diferentes preparações obtidas a partir de L. edodes e de A. blazei em patossistemas agrícolas, visando o controle de moléstias de interesse econômico como a antracnose do pepineiro, a mancha bacteriana do tomateiro e o endurecimento dos frutos do maracujazeiro. Obtida a proteção, os estudos buscaram elucidar o modo de ação das preparações de interesse, bem como purificá-las parcialmente, na tentativa de se concentrar o princípio ativo. Em plantas de pepino, extratos aquosos de basidiocarpos, obtidos a partir de diferentes isolados dos cogumelos, reduziram a severidade da antracnose, na dependência da concentração do extrato. Os extratos não afetaram adversamente o agente causal da doença, Colletotrichum lagenarium, mas provocaram o acúmulo de peroxidases e quitinases nas folhas tratadas e sistemicamente. Utilizando-se precipitação fracionada do extrato aquoso bruto de basidiocarpos de shiitake com sulfato de amônio e cromatografia de troca aniônica, obteve-se uma fração de proteínas, apresentando massa molecular de 29 a 35 kDa, com atividade elicitora de peroxidases em cotilédones de pepino. Em plantas de tomate, o isolado ABL 99/28 de A. blazei foi quem, em média, conferiu maior proteção contra Xanthomonas vesicatoria, a qual foi dependente das concentrações de extrato do cogumelo e de células bacterianas empregadas nos testes. Novamente, o extrato aquoso de basidiocarpos do isolado efetivo não atuou diretamente sobre o patógeno, mas desencadeou o aumento na atividade de b-1,3- glucanases nas folhas tratadas, sugerindo que o mecanismo de ação em pepineiro e tomateiro envolveu a indução de resistência. Já no caso do maracujazeiro, os extratos de basidiocarpos, obtidos a partir de diferentes isolados de ambos os cogumelos, protegeram localmente plantas inoculadas mecanicamente com o Passion fruit woodiness vírus (PWV), por reduzirem a infectividade viral, o que foi comprovado em testes conduzidos com Chenopodium quinoa, hospedeiro de lesão local do vírus. Entretanto, não houve proteção sistêmica em plantas de maracujá, nos experimentos de inoculação mecânica, reduzindo as possibilidades do uso dos cogumelos para o controle dessa virose no campo. De forma geral, os resultados mostraram que os cogumelos L. edodes e A. blazei apresentam compostos que ativam as respostas de defesa em plantas e podem auxiliar no controle de doenças vegetais, dependendo da natureza do agente causal. / The mushrooms Lentinula edodes and Agaricus blazei have substances in the fruiting body and in the mycelia exhibiting antibiotic activity and others able to stimulate the immune system in animals. There are many reports about the performance of these substances in the control of animal diseases. In vegetables, there are no information about the protecting effect of the royal mushroom against plant pathogens. In the case of shiitake, although few in number, the studies showed the potential of the mushroom for the control of plant diseases, such as tomato bacterial wilt, sorghum leave spots and bacterial disease of the passion fruit plant. The objectives of the present work were to evaluate the effect of different preparations from L. edodes and A. blazei to control the diseases cucumber anthracnose, tomato bacterial spot and passion fruit woodiness. As the protection of the plants was obtained, the studies tried to elucidate the way of action of the preparations, as well as partially purify them, in an attempt to concentrate the active compound. In cucumber plants, fruiting body aqueous extracts, from different mushroom isolates, reduced anthracnose severity, depending upon the extract concentration. The extracts did not affect adversely the disease causal agent, Colletotrichum lagenarium, but induced the peroxidase and quitinase accumulation in the treated leaves and systemically. By using fractional precipitation of the shiitake fruiting body aqueous extracts with ammonium sulfate, and anion exchange chromatography, a protein fraction exhibiting molecular mass around 29 to 35 kDa and peroxidase elicitor activity in cucumber cotyledons was obtained. In tomato plants, the isolate ABL 99/28 of A. blazei was the one that, on average, gave higher protection against Xanthomonas vesicatoria, which was dependent upon the extract and bacterial cell concentrations. Again, the fruiting body aqueous extract of ABL 99/28 did not act directly onto the pathogen, but it caused an increase in b-1,3-glucanase activity in the treated leaves, suggesting that the mushroom action in cucumber and tomato plants involved the induced resistance. On the other hand, the fruiting body extracts, obtained from different isolates of both mushrooms, protected locally passion fruit plants inoculated mechanically with the Passion fruit woodiness virus (PWV) by reducing viral infectivity, what was proven through tests carried out with Chenopodium quinoa, a PWV local lesion host. However, there was no systemic protection in passion fruit plants against the virus in the experiments involving mechanical inoculation, reducing the possibilities of the mushroom use for the PWV control in the field. In a general way, the results showed that the mushrooms L. edodes and A. blazei have substances that activate the plant defense mechanisms and they show some potential in the control of vegetable diseases, depending upon the nature of the pathogen.

Embora não seja reconhecido como um nutriente essencial, o silício (Si) reduz a severidade de oídios em diversas espécies vegetais quando acumulado na parte aérea. Contudo, apesar deste efeito benéfico ser amplamente relatado na literatura, ainda pouco se sabe dos mecanismos envolvidos. Este estudo avaliou o efeito das aplicações foliar e radicular de silicato de potássio, uma importante fonte de Si solúvel, na severidade de oídio e na indução de mecanismos de defesa pós-infecção em meloeiro. Os efeitos na doença foram avaliados em plantas inoculadas artificialmente ou naturalmente mantidas em casa de vegetação através da mensuração da dinâmica da epidemia e de seus componentes. Os resultados indicaram que tanto aplicações foliares como radiculares de silicato de potássio reduziram a área abaixo da curva de progresso da doença em 65% e 73%, respectivamente, comparadas ao tratamento controle sem aplicação. Tal redução foi resultado da alteração dos seguintes componentes epidemiológicos: eficiência de infecção, taxa de expansão da colônia, área da colônia, produção de conídios por área colonizada e taxa de progresso da epidemia. Contudo, a aplicação radicular foi mais eficiente que a foliar em atrasar o início da epidemia e em reduzir todos os componentes epidemiológicos, exceto a eficiência de infecção. A maior eficiência do silicato de potássio quando aplicado via raiz decorreu da maior concentração foliar do Si, a qual induziu uma antecipação (efeito priming) e um aumento da intensidade de expressão de mecanismos de defesa, fenômenos que não foram detectados no tratamento foliar. A aplicação radicular resultou na alteração de enzimas envolvidas na produção e catabolismo de espécies reativas de oxigênio, onde foi verificado aumento na atividade das enzimas superóxido dismutases e redução na atividade das enzimas catalases, principalmente nas primeiras 96 horas após a inoculação. O efeito priming também foi observado para as enzimas peroxidases, 1,3(4)-glucanases e quitinases e para o acúmulo de compostos fenólicos. Neste tratamento também ocorreu aumento na concentração de lignina em resposta à inoculação, contudo a concentração de malondialdeído foi reduzida, indicando que o Si aplicado nesta forma aliviou o estresse oxidativo sobre os lipídeos das membranas celulares da planta. Por outro lado, quando o silicato de potássio foi aplicado via foliar apenas a deposição de lignina aumentou comparado ao tratamento controle. Assim, depreende-se que os mecanismos de atuação do Si no controle da doença quando aplicado nesta forma, são distintos e envolvem, segundo a literatura, a formação de uma barreira físicoquímica sobre a cutícula resultado da polimerização do silicato de potássio. Tomados em conjunto, os resultados deste estudo evidenciaram que o Si tem papel ativo na modulação do sistema de defesa da planta, mas que sua efetividade depende de sua presença na forma solúvel no interior da planta, obtido por meio de sua absorção via raiz. Não obstante, os efeitos do silicato de potássio aqui relatados permitem concluir que a incorporação do produto a um sistema de manejo integrado, principalmente se fornecido via sistema radicular, terá efeitos positivos tanto na redução da epidemia de oídio como no meio ambiente, neste caso em função da redução no uso de fungicidas. / Silicon (Si) is not considered an essential nutrient of plants, but when it accumulates in the shoots it reduces powdery mildew severity on several species. However, despite this beneficial effect being widely reported in the literature, the mechanisms involved are still little understood. In this study, the effects of both foliar and root applications of potassium silicate, an important soluble source of Si, on the severity of powdery mildew and on the induction of postinfection defense mechanisms of melon were evaluated. Treatment effects were assessed by measuring the epidemics dynamic and its components in both artificially and naturally inoculated plants kept in the greenhouse. Results showed that the area under the disease progress curve was reduced by 65% and 73%, respectively by foliar and root application of potassium silicate, compared to control plants that were not supplied with potassium silicate. This effect accrued from the reduction of the infection efficiency, colony expansion rate, colony area, conidia production per colonized area and the epidemic progress rate. However, root application was more effective than foliar application on both delaying the onset of the epidemic and reducing most epidemic components, except for the infection efficiency. The greater efficiency of potassium silicate in controlling the disease when supplied via roots by irrigation correlated with higher foliar concentrations of Si which induced both a priming effect and increased expression of key defense responses. These phenomena, however, were not observed when potassium silicate was sprayed on leaves. Root supply of Si altered the activity of enzymes involved in the production and catabolism of reactive oxygen species, resulting in the increased activity of superoxide dismutase and reduced activity of catalases mainly in the first 96 hours after inoculation. Root application also primed the activity of peroxidases, 1,3(4)-glucanases, chitinases and the accumulation of phenolic compounds. Lignin concentration also increased in response to inoculation while the concentration of malondialdeide reduced indicating that Si decreased the oxidation of lipids of the plant cell membranes. In contrast, when Si was sprayed on the leaves, only an increase in lignin deposition was observed compared to the control treatment. Thus, it appears that the mechanisms of powdery mildew control in this case are distinct and, according to literature data, could include the formation of a physical-chemical barrier on the cuticle as the result of potassium silicate polymerization. Taken together, the results showed that Si plays an active role in modulating the host defense system, but only when present in the soluble form inside the plant which can be achieved by root uptake. Nevertheless, the effects of potassium silicate reported here allowed concluding that the inclusion of this compound in an integrated management system, mainly if supplied via roots, will render positive effects both on reducing the disease in melon plants and on the environment, in this case through reducing the use of fungicides.

Submitted by (lucia.rodrigues@ufrpe.br) on 2017-03-17T14:39:22Z No. of bitstreams: 1 Janaina Cortez de Oliveira.pdf: 616667 bytes, checksum: 9d56953508b1cd4518224b8966b15c92 (MD5) / Made available in DSpace on 2017-03-17T14:39:22Z (GMT). No. of bitstreams: 1 Janaina Cortez de Oliveira.pdf: 616667 bytes, checksum: 9d56953508b1cd4518224b8966b15c92 (MD5) Previous issue date: 2010-10-22 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPES / The bacterial blight caused by Xanthomonas citri subsp. malvacearum (Xcm) is the most important bacterial disease of cotton in the Middle-West and Northeast Regions of Brazil. The variability of a population formed by 100 Xcm strains obtained from commercial fields in the states of Goiás, Mato Grosso, Mato Grosso do Sul and Bahia, Brazil, was analyzed based upon genetic and phenotypic characteristics. The enzymes amylase, lipase, cellulase and the polysaccharide levan were produced by all Xcm isolates while the caseinase was only produced by 20 strains. All strains induced hipersensitivity reaction in tomato leaves. The 100 strains were sensitive to copper oxychloride (1500 mg L-1), copper hidroxide (1614 mg L-1), oxytetracicline (600 mg L-1), oxytetracicline (90 mg L-1) + tribasic copper sulfate (1500 mg L-1) and oxytetracicline (76.6 mg L-1) + streptomycin sulfate (367.2 mg L-1), but resistant to kasugamycin (60 mg L-1). Among the eighteen tested antibiotics the strains presented variable reaction in relation to: nalidixic acid, amoxicillin, azithromycin, bacitracin, eritromicin, kanamycin, pefloxacin, tobramycin e vancomycin. Rep-PCR (REP, ERIC and BOX) showed the formation of five groups at 70% similarity level. Group I contained 98 strains, including the reference strain Xcm IBSBF1733, group II contained only two strains from Bahia and the other three groups included strains of Xanthomonas citri pv. anacardii, X. citri pv. mangiferaeindicae e X. axonopodis pv. spondiae used for comparison. This work also evaluated the effect of silicon (Si) on the bacterial blight of cotton control and the putative mechanisms involved in the resistance potentialized by this element were evaluated. Calcium silicate (CaSiO3) was incorporated into the soil at concentrations of 0.00, 0.25, 0.50, 1.50 and 3.00 g of SiO2 kg-1 soil 25 days before planting. Leaves of 33-day-old-plants were inoculated by infiltration with 0.5 mL suspension of Xcm (108 CFU ml-1). Components of resistance were evaluated every two days until 10 days after inoculation, at which point plant development and the accumulation of Si and Calcium (Ca) were also determined. H2O2 production and the activity of enzymes related to plant defenses were analyzed at 6, 12 and 24 hours after inoculation in plants +/-Si. In vitro inhibition of pathogen growth due to Si was also assessed. No significant difference was seen among the treatments regarding incubation period, disease incidence or inhibition of bacterial growth. However, with the application of 1.50 g of SiO2 kg-1 of soil, a reduction in severity (54.9%) was observed along with a reduction in the area under the disease progression curve (35.76%) and an increase in plant height (7.04%). There was no observable accumulation of Si in the cotton leaves. Levels of soluble proteins and H2O2 and activity of the enzymes SOD, APX, guaiacol peroxidase, PAL and b Glu were altered in the presence of Si (1.80 g SiO2 kg-1). Thus based on the genetic and phenotypic characteristics studied we conclude that the population of 100 Xcm strains presents low variability, and that the reduction of cotton bacterial blight severity mediated by Si is probably associated to the characteristic events of induced resistance observed. / A mancha-angular, causada por Xanthomonas citri subsp. malvacearum (Xcm), é a principal doença bacteriana do algodoeiro no Centro-Oeste e Nordeste do Brasil. A variabilidade de uma população de 100 isolados de Xcm, oriundos de plantas com sintomas típicos de mancha-angular em áreas produtoras de algodoeiro dos estados de Goiás, Mato Grosso, Mato Grosso do Sul e Bahia, Brasil, foi analisada com base em características fenotípicas e genéticas. As enzimas amilase, lipase e celulase e o polissacarídeo levana foram produzidos por todos os isolados de Xcm, enquanto a caseinase foi produzida apenas por 20 isolados. Todos os isolados induziram reação de hipersensibilidade em folhas de tomateiro. Os 100 isolados foram sensíveis a oxicloreto de cobre (1500 mg l-1), hidróxido de cobre (1614 mg l-1), oxitetraciclina (600 mg l-1), oxitetraciclina (90 mg l-1) + sulfato tribásico de cobre (1500 mg l-1) e oxitetraciclina (76,6 mg l-1) + sulfato de estreptomicina (367,2 mg l-1), e resistentes a casugamicina (60 mg l-1). Dentre os 18 antibióticos testados, os isolados bacterianos apresentaram reação variável a nove: ácido nalidíxico, amoxicilina, azitromicina, bacitracina, eritromicina, kanamicina, pefloxacina, tobramicina e vancomicina. Rep-PCR (REP, ERIC e BOX) revelou a formação de cinco grupos ao nível de 70% de similaridade, sendo o grupo I constituído de 98 isolados, incluindo o isolado referência de Xcm IBSBF1733, o grupo II, formado apenas por dois isolados oriundos da Bahia e os três outros grupos por isolados de X. citri pv. anacardii, X. citri pv. mangiferaeindicae e X. axonopodis pv. spondiae utilizados para comparação. Neste trabalho também foi avaliado o efeito do silício (Si) no controle da mancha-angular do algodoeiro, e os possíveis mecanismos envolvidos na resistência potencializada por este elemento. O silicato de cálcio (CaSiO3) foi incorporado ao solo nas doses de 0,00; 0,25; 0,50; 1,50 e 3,00 g de SiO2 kg-1 de solo 25 dias antes do plantio. Plantas com 33 dias tiveram as folhas inoculadas por infiltração com 0,5 mL de suspensão de Xcm (108 UFC ml-1). Os componentes da resistência foram avaliados a cada dois dias até 10 dias após a inoculação, quando o desenvolvimento da planta e o acúmulo de Si e Ca foram também determinados. Foram ainda analisadas a atividade de enzimas relacionadas à defesa vegetal e a produção de H2O2 às 6, 12 e 24 h após a inoculação em plantas +/-Si e a inibição in vitro do crescimento do patógeno. Não foi verificada diferença significativa entre os tratamentos quanto ao período de incubação, incidência da mancha-angular e inibição do crescimento da bactéria. Com a aplicação da dose 1,50 g SiO2 kg-1 de solo, observou-se redução da severidade (54,9%), da área abaixo da curva de progresso da doença (35,76%) e aumento na altura das plantas (7,04%). Não foi verificado acúmulo de Si nas folhas de algodoeiro. Níveis de proteínas solúveis e H2O2 e atividade das enzimas SOD, APX, POX, PAL e GLU foram alterados na presença do Si (1,80 g SiO2 kg-1). Com base nas características fenotípicas e genéticas analisadas, conclui-se que a população de 100 isolados de Xcm estudada apresenta baixa variabilidade e que a redução da severidade da mancha-angular do algodoeiro mediada por Si está provavelmente associada aos eventos característicos de indução de resistência observados.

This thesis is an investigation of detecting and quantifying airborne inoculum of Sclerotinia sclerotiorum (Lib.) de Bary to improve the Sclerotinia rot of carrot (SRC) forecast model. A quantitative polymerase chain reaction (qPCR) assay was developed to specifically detect and quantify DNA from airborne ascospores of S. sclerotiorum. The qPCR assay was evaluated on air samples collected using a Burkard Sampler, and showed that ascospores of S. sclerotiorum were specifically detected among a pool of foreign DNA. The concentration of detected ascospores was related to the observed incidence of SRC to suggest a preliminary threshold of 2 to 4 ascospores m-3 of air for SRC development. Evaluation of an Andersen Sampler, the blue plate test (BPT) and the qPCR assay showed that the latter two methods were equally effective in detecting and quantifying ascospores of S. sclerotiorum and consistently detected greater numbers of ascospores than an Andersen Sampler. Three days are required to confirm the presence of S. sclerotiorum using the BPT, while results from the qPCR assay can potentially provide results within five hours of air sampling. The choice of detection method depends on the available resources and need for a quick result. Analysis of data from nine years of air sampling using the BPT indicated that a single air sampling site is sufficient to detect ascospores when counts are low, increasing to two sites during periods when ascospores are detected near threshold levels and crop and environmental conditions are conducive to disease. Chitosan and canopy trimming were evaluated to manage SRC under field conditions. Chitosan reduced area under the disease progress curve (AUDPC) by 55 and 42% in 2009 and 2011, respectively, which was comparable to a standard fungicide. Trimming enhanced chitosan efficacy, reducing AUDPC by 88 and 82% in 2009 and 2011, respectively. Trimming as a stand-alone treatment reduced AUDPC by 66% in 2011. Under controlled environmental conditions, chitosan inconsistently enhanced defense responses against S. sclerotinia. The results show that chitosan has potential to be integrated into SRC management systems, particularly when combined with foliar trimming in years with moderate to high disease risk. / National Research Council of Canada; University of Guelph; Department of Plant Agriculture; Ontario Ministry of Agriculture, Food and Rural Affairs

Hormesis is a dose response phenomenon where low doses of a stress bring about a positive response in the organism undergoing treatment. UV-C hormesis has been known for over three decades and has a broad range of benefits on postharvest produce. Benefits include increased nutritional content, delayed chlorophyll degradation and disease resistance. The beneficial effects have been observed on many varieties of fresh produce including climacteric and non-climacteric fruit, tubers, salads and brassicas. The majority of previous studies have used low-intensity (LIUV) UV-C sources. LIUV sources require lengthy treatment times, which are in the region of 6 minutes for tomato fruit. This has, in part, prevented the commercial application of this technique. High-intensity, pulsed polychromatic light (HIPPL) sources, however, have recently been developed. HIPPL sources may have the potential to drastically reduce treatment times and increase their commercial viability. It was shown, here, that the use of HIPPL can control disease (reduce disease progression) caused by Botrytis cinerea and Penicillium expansum and also delay ripening on tomato fruit. Both disease control and delayed ripening were at similar levels for LIUV and HIPPL treatments on mature green fruit. The HIPPL treatments used in these studies can reduce treatment times for tomato fruit by 97.3%. Both HIPPL and LIUV treatments elicit local responses irrespective of the treatment orientation and tomato fruit, therefore, require full surface irradiation. Furthermore, UV-C in the HIPPL source is not required for disease control or delayed ripening. It does, however, contribute approximately 50% towards the total observed effects. Investigations into the mechanisms underpinning postharvest HIPPL and LIUV hormesis, on tomato fruit, identified that the expression of genes involved in plant hormone biosynthesis, defence, secondary metabolism and ripening were affected. This indicates that disease control is achieved through induced resistance. Changes to expression, following treatment, were highly similar for both HIPPL and LIUV treatments and were mediated by salicylic acid, jasmonic acid and ethylene. This may lead to broad range resistance against necrotrophic and biotrophic pathogens as well as abiotic stresses and herbivorous pests. Recently, the exposure of foliage to UV-C has been shown to induce resistance against B. cinerea on Arabidopsis thaliana. The horticultural applications of such treatments, however, have not been explored. Pre-harvest treatments of lettuce in the glasshouse showed variation in damage threshold and optimal treatment to control disease following LIUV and HIPPL treatment. Further sources of variation included the cultivar, pathogen of interest and the point that treatment was applied during the year. Using a controlled environment allowed seasonal variation to be mitigated and both HIPPL and LIUV treatments controlled disease against B. cinerea. For pre-harvest treatments to be a success in the glasshouse, further studies into how both biotic and abiotic factors influence treatment is required. To circumvent the problems associated with pre-harvest treatments and environmental variation in the glasshouse, LIUV seed treatments were performed on tomato. Control of B. cinerea was established with an approximately 10% reduction in incidence and disease progression with a 4 kJ/m2 treatment. When monitoring the effect of treatment on germination and early seedling development it was also identified that an 8 kJ/m2 treatment led to biostimulation of germination and root and shoot growth.

Submitted by Erika Demachki (erikademachki@gmail.com) on 2014-11-06T18:02:43Z No. of bitstreams: 2 Dissertação - Camila de Souza Queiroz - 2012.pdf: 913358 bytes, checksum: eee67ccbeb48729433c4d52ca1da5433 (MD5) license_rdf: 23148 bytes, checksum: 9da0b6dfac957114c6a7714714b86306 (MD5) / Approved for entry into archive by Erika Demachki (erikademachki@gmail.com) on 2014-11-06T18:03:27Z (GMT) No. of bitstreams: 2 Dissertação - Camila de Souza Queiroz - 2012.pdf: 913358 bytes, checksum: eee67ccbeb48729433c4d52ca1da5433 (MD5) license_rdf: 23148 bytes, checksum: 9da0b6dfac957114c6a7714714b86306 (MD5) / Made available in DSpace on 2014-11-06T18:03:27Z (GMT). No. of bitstreams: 2 Dissertação - Camila de Souza Queiroz - 2012.pdf: 913358 bytes, checksum: eee67ccbeb48729433c4d52ca1da5433 (MD5) license_rdf: 23148 bytes, checksum: 9da0b6dfac957114c6a7714714b86306 (MD5) Previous issue date: 2012-06-29 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPES / An alternative for managing the disease might be performing plant nutrition since mineral nutrients are directly involved in all mechanisms of the plant’ defense system as integral components of cells, membranes, enzymes and electron transport or by activation, inhibition and regulation of metabolism. In this context the study aimed at: 1- assessing the influence of soybean nutrition with micronutrients and Si in the severity of Asian soybean rust in field conditions and a greenhouse experiment; 2- evaluating the influence of soybean nutrition with different doses and ways of Ni application in the severity of Asian soybean rust in greenhouse conditions; 3- assessing fungal toxicity of the nickel sulfate for P. pachyrhizi spores in laboratory conditions. Results showed rust severity reduction in treatments with foliar application of Zn, Si and Ni, which promoted a reduction in the rate of the disease progression in the field. The grain yield was not affected by nutrition in the field; however, the treatments with Zn applications increased grain production in the greenhouse. On the other hand, the treatments with Si application reduced the disease severity although there was a grain yield reduction in the greenhouse. The Ni promoted a lower rust severity and there was an increase in the grain yield in treatments with applications of doses 20 to 40 g ha-1. Nevertheless, the rate of the progression of the disease was not reduced in the treatments with Ni. The treatments with fungicide reduced the severity of the disease, but the grain yield did not increase. In laboratory conditions the nickel sulfate reduced the germination in vitro of P. pachyrhizi spores. / Uma alternativa de manejo da ferrugem-asiática (Phakopsora pachyrhizi) na soja é a nutrição adequada das plantas, uma vez que os nutrientes minerais são diretamente envolvidos em mecanismos de defesa, como componentes integrais das células, das membranas, das enzimas e dos transportadores de elétrons ou como ativadores, inibidores e reguladores do metabolismo. Neste contexto, objetivou-se através deste trabalho avaliar a influência de micronutrientes (B, Zn, Mn e Cu), silício e níquel na severidade da ferrugem-asiática na cultura da soja em casa de vegetação, campo e laboratório. Os resultados mostraram redução da severidade da ferrugem nos tratamentos com aplicação foliar de Zn, Si e Ni os quais promoveram redução da taxa de progresso da doença no experimento de campo. A produção de grãos não foi influenciada pela nutrição no experimento em campo, entretanto, os tratamentos com Zn aplicado via foliar aumentaram a produção de grãos em casa-de-vegetação. Os tratamentos com aplicação de Si reduziram a severidade da doença, porém reduziram a produção de grãos em casa de vegetação. O sulfato de Ni promoveu menor severidade da ferrugem e incremento na produção de grãos nos tratamentos com aplicação de doses 20 – 40 g ha-1. No entanto, os tratamentos com Ni não reduziram a taxa de progresso da doença. Os tratamentos com fungicida reduziram a severidade da doença, porém não influenciaram a produção de grãos. Em laboratório, o sulfato de níquel reduziu a germinação in vitro dos esporos de P. pachyrhizi.

Submitted by Cláudia Bueno (claudiamoura18@gmail.com) on 2016-06-02T18:09:18Z No. of bitstreams: 2 Dissertação - Rafaela Araújo Guimarães - 2014.pdf: 1551849 bytes, checksum: b39011c17618c122fad17d142b0288ec (MD5) license_rdf: 23148 bytes, checksum: 9da0b6dfac957114c6a7714714b86306 (MD5) / Approved for entry into archive by Luciana Ferreira (lucgeral@gmail.com) on 2016-06-03T12:41:12Z (GMT) No. of bitstreams: 2 Dissertação - Rafaela Araújo Guimarães - 2014.pdf: 1551849 bytes, checksum: b39011c17618c122fad17d142b0288ec (MD5) license_rdf: 23148 bytes, checksum: 9da0b6dfac957114c6a7714714b86306 (MD5) / Made available in DSpace on 2016-06-03T12:41:12Z (GMT). No. of bitstreams: 2 Dissertação - Rafaela Araújo Guimarães - 2014.pdf: 1551849 bytes, checksum: b39011c17618c122fad17d142b0288ec (MD5) license_rdf: 23148 bytes, checksum: 9da0b6dfac957114c6a7714714b86306 (MD5) Previous issue date: 2014-08-21 / Conselho Nacional de Pesquisa e Desenvolvimento Científico e Tecnológico - CNPq / Sarocladium oryzae, the causal agent of rice sheath rot disease is described as antagonistic to rice pathogens. Rice blast is a major rice disease and is responsible for losses up to 100% in productivity. The disease control is done by integrated management, where the main practices are use of resistant cultivars and chemical control. The biocontrol agents or their metabolites may to be more practical to be including them as components in the management. The in objectives of this study a consist to evaluate of S. oryzae isolates for the morphological variability, genetics, biochemistry and antagonistic activity to rice pathogens; evaluate effect of S. oryzae filtrade on conidial germination and appressorium formation of M. oryzae; evaluate potential of conidia and filtrate S. oryzae in the suppression of leaf blast severity and quantify activity of enzymes involved in interaction M. oryzae x rice plant x S. oryzae. Isolates were characterized for color, texture, colony diameter, conidia size and hyphae thickness. In genetic studies, we used RAPD marker primers, and cerulenin production was quantified by HPLC. Antagonism in vitro was assessed by dual culture method. The effect of S. oryzae on conidial germination and appressorium formation of M. oryzae was evaluated using hydrophobic surface. Rice cultivar BRS Primavera, M. oryzae isolate (Py 10.900) and S. oryzae, isolate So 03, were utilized to study plant-pathogen-antagonist relationship. Plants were sprayed, with conidial suspension (CS), 3x105 conídios.mL-1 and culture filtrate (CF) 100% of S. oryzae, two days before inoculation with M. oryzae. S. oryzae isolates showed morphological variability, polymorphism in DNA. A majority of S. oryzae isolates (60%) ware able to produce cerulenin and over 55% were antagonistic to C. miyabeans, M. oryzae, M. albescens and T. cucumeris. The isolate So 29 showed largest inhibition zone. Filtrate of isolates So 03 and So 29 delayed conidia germination by 89.5% and inhibited appressoria formation of M. oryzae by 85%. CS reduced of leaf blast severity in 68.8% and CF in 75.5%. The enzymes β-1,3-glucanase and peroxidase exhibited maximum activity in plants sprayed with CF, when as the activity was high for SA and lipoxygenase in relation to CS treatment, compared to their respective controles, in the absence of M. oryzae. After inoculation with M. oryzae, the lipoxygenase and phenylalanine ammonia-lyase activity in both treatments CF and CS, showed differences compared to controls (plants inoculated with M. oryzae and water only). S. oryzae presented variability to characteristics evaluated and showed potential antagonism against to rice pathogens. Changes in enzyme activity indicate their role in induction resistance in plants in M. oryzae x rice x S. oryzae interaction. / Sarocladium oryzae, agente causal da podridão da bainha do arroz, é descrito como antagonista a patógenos de arroz. A brusone, principal doença do arroz é responsável por perdas de até 100% na produtividade. O controle desta doença é feito pelo uso do manejo integrado, onde as principais práticas são o uso de cultivares resistentes e o controle químico. O uso de agentes de biocontrole ou seus metabólitos pode ser mais uma prática a ser inserida ao manejo. Os objetivos deste estudo foram: avaliar isolados de S. oryzae quanto à variabilidade morfológica, genética, bioquímica e quanto à atividade antagônica aos patógenos de arroz; avaliar o efeito do filtrado de S. oryzae na germinação de conídios e na formação de apressórios de M. oryzae; avaliar o potencial da suspensão de conídios e do filtrado de S. oryzae na supressão da brusone foliar e quantificar a atividade das enzimas relacionadas à patogênese e do fitohormônio ácido salicílico (AS) envolvidos na interação M. oryzae x arroz x S. oryzae. Os isolados foram caracterizados quanto à cor, textura, diâmetro da colônia, tamanho dos conídios e espessura das hifas. No estudo genético, utilizou-se marcador RAPD (Random Amplified Polymorphic DNA) e a produção de cerulenina foi quantificada em HPLC. O antagonismo in vitro foi avaliado pelo método da cultura pareada. A ação de S. oryzae sobre a germinação de conídios e a formação de apressórios de M. oryzae foi avaliada em superfície hidrofóbica. Plantas de arroz da cultivar BRS Primavera, um isolado virulento de M. oryzae (Py 10.900) e o isolado So 03 de S. oryzae, foram avaliados no estabelecimento das relações planta-patógeno-antagonista. As plantas foram pulverizadas com S. oryzae, na forma de suspensão de conídios (SC) - 3x105 conídios.mL-1 e filtrado (FI) 100% concentrado, dois dias antes da inoculação com M. oryzae. Os isolados de S. oryzae apresentaram variabilidade morfológica e polimorfismo no DNA. A maioria dos isolados de S. oryzae (60%) foi capaz de produzir cerulenina e mais de 55% foram antagônicos a C. miyabeans, M. oryzae, M. albescens e T. cucumeris. O isolado So 29 apresentou o maior halo de inibição no pareamento. Os filtrados dos isolados So 03 e So 29 retardaram a germinação dos conídios em 89,5% e inibiram a formação dos apressórios de M. oryzae em 85%. A SC reduziu a severidade da brusone foliar em 68,8% e o FI em 75,5%. Os maiores valores de atividade enzimática específica em relação ao controle antes da presença de M. oryzae foram para β-1,3-glucanase e peroxidase no tratamento com FI, enquanto que, na SC foram lipoxigenase e AS. Depois da presença de M. oryzae a lipoxigenase e a fenilalanina-amônia liase apresentaram atividade tanto com FI quanto na SC, diferindo dos controles (plantas inoculadas com água e com M. oryzae, somente). S. oryzae apresentou variabilidade para as características avaliadas e potencial antagônico aos patógenos do arroz. As alterações na atividade enzimática indicam a indução de resistência em plantas na interação M. oryzae x arroz x S. oryzae.

Os cogumelos Lentinula edodes (shiitake) e Agaricus blazei (cogumelo-do-sol) apresentam substâncias no corpo de frutificação (basidiocarpo) e no micélio com atividades antibióticas e imuno-regulatórias, havendo uma série de relatos sobre a atuação das mesmas no controle de doenças em animais. Em vegetais, não há informações sobre o efeito protetor do cogumelo-do-sol contra fitopatógenos. No caso de shiitake, embora pouco numerosos, os estudos mostraram o potencial do cogumelo para o controle de doenças de plantas, tais como a murcha bacteriana do tomateiro, a murcha de feijão-lima, além de doenças fúngicas em sorgo e da bacteriose do maracujazeiro. Os objetivos do presente trabalho foram o de avaliar o efeito de diferentes preparações obtidas a partir de L. edodes e de A. blazei em patossistemas agrícolas, visando o controle de moléstias de interesse econômico como a antracnose do pepineiro, a mancha bacteriana do tomateiro e o endurecimento dos frutos do maracujazeiro. Obtida a proteção, os estudos buscaram elucidar o modo de ação das preparações de interesse, bem como purificá-las parcialmente, na tentativa de se concentrar o princípio ativo. Em plantas de pepino, extratos aquosos de basidiocarpos, obtidos a partir de diferentes isolados dos cogumelos, reduziram a severidade da antracnose, na dependência da concentração do extrato. Os extratos não afetaram adversamente o agente causal da doença, Colletotrichum lagenarium, mas provocaram o acúmulo de peroxidases e quitinases nas folhas tratadas e sistemicamente. Utilizando-se precipitação fracionada do extrato aquoso bruto de basidiocarpos de shiitake com sulfato de amônio e cromatografia de troca aniônica, obteve-se uma fração de proteínas, apresentando massa molecular de 29 a 35 kDa, com atividade elicitora de peroxidases em cotilédones de pepino. Em plantas de tomate, o isolado ABL 99/28 de A. blazei foi quem, em média, conferiu maior proteção contra Xanthomonas vesicatoria, a qual foi dependente das concentrações de extrato do cogumelo e de células bacterianas empregadas nos testes. Novamente, o extrato aquoso de basidiocarpos do isolado efetivo não atuou diretamente sobre o patógeno, mas desencadeou o aumento na atividade de b-1,3- glucanases nas folhas tratadas, sugerindo que o mecanismo de ação em pepineiro e tomateiro envolveu a indução de resistência. Já no caso do maracujazeiro, os extratos de basidiocarpos, obtidos a partir de diferentes isolados de ambos os cogumelos, protegeram localmente plantas inoculadas mecanicamente com o Passion fruit woodiness vírus (PWV), por reduzirem a infectividade viral, o que foi comprovado em testes conduzidos com Chenopodium quinoa, hospedeiro de lesão local do vírus. Entretanto, não houve proteção sistêmica em plantas de maracujá, nos experimentos de inoculação mecânica, reduzindo as possibilidades do uso dos cogumelos para o controle dessa virose no campo. De forma geral, os resultados mostraram que os cogumelos L. edodes e A. blazei apresentam compostos que ativam as respostas de defesa em plantas e podem auxiliar no controle de doenças vegetais, dependendo da natureza do agente causal. / The mushrooms Lentinula edodes and Agaricus blazei have substances in the fruiting body and in the mycelia exhibiting antibiotic activity and others able to stimulate the immune system in animals. There are many reports about the performance of these substances in the control of animal diseases. In vegetables, there are no information about the protecting effect of the royal mushroom against plant pathogens. In the case of shiitake, although few in number, the studies showed the potential of the mushroom for the control of plant diseases, such as tomato bacterial wilt, sorghum leave spots and bacterial disease of the passion fruit plant. The objectives of the present work were to evaluate the effect of different preparations from L. edodes and A. blazei to control the diseases cucumber anthracnose, tomato bacterial spot and passion fruit woodiness. As the protection of the plants was obtained, the studies tried to elucidate the way of action of the preparations, as well as partially purify them, in an attempt to concentrate the active compound. In cucumber plants, fruiting body aqueous extracts, from different mushroom isolates, reduced anthracnose severity, depending upon the extract concentration. The extracts did not affect adversely the disease causal agent, Colletotrichum lagenarium, but induced the peroxidase and quitinase accumulation in the treated leaves and systemically. By using fractional precipitation of the shiitake fruiting body aqueous extracts with ammonium sulfate, and anion exchange chromatography, a protein fraction exhibiting molecular mass around 29 to 35 kDa and peroxidase elicitor activity in cucumber cotyledons was obtained. In tomato plants, the isolate ABL 99/28 of A. blazei was the one that, on average, gave higher protection against Xanthomonas vesicatoria, which was dependent upon the extract and bacterial cell concentrations. Again, the fruiting body aqueous extract of ABL 99/28 did not act directly onto the pathogen, but it caused an increase in b-1,3-glucanase activity in the treated leaves, suggesting that the mushroom action in cucumber and tomato plants involved the induced resistance. On the other hand, the fruiting body extracts, obtained from different isolates of both mushrooms, protected locally passion fruit plants inoculated mechanically with the Passion fruit woodiness virus (PWV) by reducing viral infectivity, what was proven through tests carried out with Chenopodium quinoa, a PWV local lesion host. However, there was no systemic protection in passion fruit plants against the virus in the experiments involving mechanical inoculation, reducing the possibilities of the mushroom use for the PWV control in the field. In a general way, the results showed that the mushrooms L. edodes and A. blazei have substances that activate the plant defense mechanisms and they show some potential in the control of vegetable diseases, depending upon the nature of the pathogen.