O fungo Sclerotinia sclerotiorum, agente causal do mofo branco, encontra-se disseminado em todo o país e afeta mais de 400 espécies de plantas, dentre elas a cultura da soja, causando danos consideráveis no rendimento. A transmissão do patógeno ocorre por meio de sementes contendo micélio dormente ou pela presença de escleródios no lote, os quais podem permanecer viáveis por um longo período, dificultando o manejo da doença. Este trabalho teve como objetivos verificar a viabilidade do patógeno em sementes durante o armazenamento e seu efeito sobre o poder germinativo; comparar tamanhos de amostras de sementes visando à detecção do fungo; e avaliar a sensibilidade in vitro de S. sclerotiorum a fungicidas. Para avaliação da sobrevivência do patógeno, sementes de soja foram inoculadas, artificialmente, com três isolados de S. sclerotiorum, e armazenadas em duas condições, câmara fria e seca (20 ºC e 45% UR) e ambiente não controlado, durante oito meses. A cada dois meses foram realizados testes de sanidade pelo meio de cultura ágar-bromofenol (Neon), utilizando-se 4 repetições de 100 sementes, e testes de germinação, com 4 repetições de 50 sementes, pelo método do rolo de papel. No experimento de amostragem foram avaliadas dez amostras de sementes de soja pelo método do meio Neon, comparando-se quatro tamanhos de amostra: 400, 800, 1600 e 2400 sementes. Para avaliação da sensibilidade in vitro de S. sclerotiorum a fungicidas, foram utilizados 13 ingredientes ativos, em 7 doses e 5 repetições. Os valores da concentração inibitória de 50% (CI50) e 100% (CI100) foram estimados com base na percentagem de inibição do crescimento micelial. Além disso, realizaram-se a quantificação e pesagem dos escleródios formados após 15 dias de incubação. A presença do patógeno S. sclerotiorum, afetou negativamente a germinação das sementes, com redução média de 32% em relação à testemunha. Verificou-se diferença entre os isolados com relação à agressividade. O patógeno sobreviveu nas sementes em ambos os ambientes de armazenamento, porém, em ambiente controlado a redução na geminação das sementes foi de 14% e na incidência do fungo de 16%, enquanto que, em laboratório, esta redução foi de 23% na germinação e de 62% na incidência. Não houve diferença estatística com relação à quantidade de sementes amostradas na detecção de S. sclerotiorum, entretanto, constatou-se maior frequência de ocorrência do patógeno à medida que o número de sementes amostrado foi maior. Todos os fungicidas foram capazes de reduzir o crescimento micelial de S. sclerotiorum, bem como a produção de escleródios, sendo que os ingredientes ativos piraclostrobina, carbendazim, tiabendazol e tiofanato metílico, além de apresentarem CI50 inferior a 1 mg L-1, também inibiram totalmente a formação de escleródios em concentrações inferiores a 5 mg L-1. / The fungus Sclerotinia sclerotiorum, the causal agent of white mold is spread throughout the country and affects more than 400 species of plants, including soybean, causing considerable decrease in yield. The pathogen transmission occurs through seeds with dormant mycelium or by the presence of sclerotia in the lot, which can remain viable for a longer period, making hard the disease management. This study aimed to verify the viability of the pathogen in seeds during the storage period and its effect on germinative power; compare sizes of seed samples to determine the presence of the fungus; and to evaluate the in vitro sensibility of S. sclerotiorum to fungicides. To determine the pathogen survival, soybean seeds were inoculated artificially with three isolates of S. sclerotiorum, and stored into two conditions, cold and dry chamber (20 °C and 45% RU) and in uncontrolled environment, for eight months. Every two months health tests were performed by culture media bromophenol agar (Neon), using 4 repetition of 100 seeds, and germination tests, with 4 repetition of 50 seeds, by the method of the paper roll. During the experiment of sampling, ten samples of soybean seeds were evaluated by the method of media Neon, comparing four sample sizes: 400, 800, 1600 and 2400 seeds. To evaluate the in vitro sensibility of S. Sclerotinia to fungicides were used 13 active ingredients, in 7 doses and 5 repetitions. The values of the 50% inhibitory concentration (IC50) and 100% (IC100) were estimated based on the percentage inhibition of the mycelial growth. In addition, it was performed the quantification and weighing of sclerotia formed after 15 days of incubation. The pathogen S. sclerotiorum in soybean seeds affected negatively the germination of seeds, with an average reduction of 32% compared to the control. There was difference between the isolates in relation to aggressiveness. The pathogen survived in seeds in both storage environments, however, under controlled environment, the decrease in the germination of the seeds was 14% and in its incidence was 16%, while in laboratory this reduction was 23% in the germination and 62% in the incidence. There was no statistical difference in the quantity of seeds sampled in the detection of S. sclerotiorum, however, a higher frequency of pathogen occurrence was verified as the number of sampled seeds was enlarged. All fungicides were able to reduce the mycelial growth of S. sclerotiorum and sclerotia production, and the active ingredients pyraclostrobin, carbendazim, thiabendazole and thiophanate methyl, besides having IC50 of less than 1 mg L-1 also completely inhibited the formation of sclerotia in concentrations less than 5 mg L-1.

O controle da podridão parda do pessegueiro (Monilinia fructicola) é realizado essencialmente por aplicações preventivas de fungicidas, em sua maioria sistêmicos com modo de ação específico. O uso contínuo desses fungicidas pode levar à seleção de indivíduos resistentes ao ingrediente ativo e, consequentemente, à perda da eficiência do controle nos pomares. A evolução da população de isolados resistentes ao longo do tempo, na ausência do fungicida, pode ser prevista com estudos de habilidade competitiva entre isolados resistentes e sensíveis. Portanto, a quantificação da adaptabilidade (fitness) e da estabilidade, necessárias para caracterizar os isolados encontrados em campo, auxiliarão nas estratégias antirresistência. Embora a importância da determinação da adaptabilidade de isolados resistentes seja reconhecida, há poucos métodos descritos para a análise da habilidade competitiva. Assim, esse trabalho verificou a adaptabilidade, a estabilidade e a habilidade competitiva de isolados de M. fructicola com diferentes níveis de sensibilidade à azoxistrobina em experimentos in vitro e em frutos de pêssegos (in natura e enlatado) utilizando metodologias de avaliação diversas. A caracterização dos isolados foi realizada in vitro, in vivo e molecularmente. Foram avaliados o diâmetro da colônia e a esporulação in vitro; o período de incubação e de latência, a incidência e a severidade da doença e a esporulação in vivo dos isolados separadamente. Os dados de esporulação in vitro foram submetidos ao teste de Kruskal-Wallis e os dados de crescimento micelial ao teste de Tukey com 5% de probabilidade. Para a análise dos dados in vivo, utilizou-se a análise multivariada. Dez transferências em meio de cultura sem fungicida foram realizadas para o estudo da estabilidade de dois isolados resistentes (SP08435 e SP09839) e um sensível (PR09638). A esporulação, a porcentagem de germinação de conídios e o diâmetro da colônia dos isolados em meio de cultura com fungicida foram avaliados em quatro transferências (inicial, terceira, sexta e última transferência). A habilidade competitiva foi verificada pela evolução da porcentagem de indivíduos resistentes no tempo entre um isolado resistente (SP09839) e um sensível à azoxistrobina (PR09638). Modelos lineares ou modelos lineares generalizados (GLM) foram utilizados para descrever o progresso da proporção do isolado resistente ao longo do tempo. Os isolados resistentes exibiram esporulação in vitro e taxa de crescimento micelial semelhantes aos sensíveis, além de apresentarem-se no mesmo grupo de similaridade que os isolados sensíveis. Nenhuma mutação no cyt b foi associada à resistência de M. fructicola à azoxistrobina (G143A, G137R, e F129L) e em todos os isolados foi observado o íntron antes do códon 143. Mesmo após 10 transferências em meio de cultura sem fungicida, os isolados resistentes à azoxistrobina não reduziram sua sensibilidade nem sua capacidade de esporulação e crescimento fora do pêssego. Independentemente da metodologia utilizada para avaliar a habilidade competitiva, a resistência à azoxistrobina não correspondeu a um custo adaptativo para M. fructicola, pois a proporção do isolado resistente manteve-se inalterada ao longo das transferências. O uso de pêssego enlatado é recomendado na substituição de frutos in natura para experimentos de habilidade competitiva. / Control of brown rot in peaches (caused by Monilinia fructicola) is mainly by preventive applications of fungicides, mostly single-target site. Repeated use of these fungicides can, however, select for resistant pathogen populations, which may result in failure of disease control. The evolution of resistant isolates in a population over time can be predicted with competitive ability assays between sensitive and resistant isolates. Therefore, studies quantifying pathogen fitness and stability are necessary to characterize field isolates and improve anti-resistant strategies. However, there are limited reports concerning the methodology used in competitive ability assays. This study determined the fitness of M. fructicola isolates with different levels of sensitivity to azoxystrobin in trials in vitro and in vivo (in natura and canned peaches) with different monocyclic components. Isolates characterization were carried out in vitro, in vivo and molecularly. Mycelial growth rate and sporulation were estimated in vitro, and disease incidence, severity, lesion sporulation, incubation and latent period were evaluated in peach fruit for sensitive and resistant isolates. Sporulation in vitro data were analyzed by Kruskal-Wallis test and mycelial growth rate with the Tukey test at 5% probability. For the in vivo experiments, multivariate analysis were carried out. Ten consecutive transfers on non-amended media were conducted to determine the stability of isolates (two resistant and one sensitive). Sporulation, percent spore germination and colony diameter on azoxystrobin amended media were also evaluated four times (the first, the third, the sixth and the last transfer). Competitive ability was investigated by testing the percentage increase of resistant individuals five generations after inoculation with a mix of azoxystrobin sensitive and resistant isolates. The competitive ability of sensitive and resistant isolates were compared with different variables as colony-forming unit and spore germination. Linear or generalized linear models (GLM) were adjusted to compare the progress of resistant isolates over time. All resistant isolates had in vitro sporulation and mycelial growth rate similar to sensitive isolates. All isolates sequenced presented an intron close to codon 143 and did not have any of the point mutations commonly associated with resistance to QoI fungicides (G143A, G137R, and F129L). After 10 transfers in PDA, colony diameter of resistant isolates was not reduced in azoxystrobin-amended media with 1 μg.ml-1 nor was their sporulation in vitro For all parameters used to measure competitive ability, no fitness cost was associated to M. fructicola isolates resistant to azoxystrobin. Canned peach is a viable option for studies aiming to determine fitness cost of resistant isolates.

A utilização de fungicidas multissítios, como é o caso do mancozebe (Mz), tem crescido bastante nas últimas safras com o intuito de evitar a resistência de fungos a fungicidas sítio-específico, além de aumentar sua eficiência no controle de doenças. No entanto, além do efeito fungicida, em áreas que receberam a aplicação destes produtos tem sido observados efeitos fisiológicos nas plantas, que vão desde a redução do nível de estresse até o aumento no teor de clorofila, o que repercute na produtividade. Associado a isto, o ácido salicílico também se torna uma estratégia importante, visto que este hormônio promove redução do nível de estresse, além de atuar na indução de resistência contra doenças. No entanto, pouco se sabe sobre os efeitos fisiológicos que este produtos isolados e em associação podem causar na cultura do feijoeiro comum. Portanto, o objetivo do trabalho foi avaliar o efeito da aplicação de Mz e ácido salicílico em características fisiológicas, componentes de produção e produtividade da cultura do feijoeiro comum. O ensaio foi conduzido em área comercial de produção de feijão irrigado em Patos de Minas-MG entre os meses de abril e agosto de 2017. Foi adotado delineamento em blocos casualizados em esquema fatorial 5 x 2 constituído por cinco doses de ácido salicílico (0, 50, 100, 200 e 400 mg ha-1) associados ou não a aplicação de Mz na dose de 400 g ha-1 e cinco repetições. Para cada tratamento, foram realizadas três aplicações, sendo estas aos 22, 37 e 52 dias após a semeadura. Foram realizadas avaliações de massa de matéria seca de vagens, produtividade, atividade da nitrato redutase, urease, enzimas antioxidantes (SOD, POD e CAT), teor de prolina, peróxido de hidrogênio e peroxidação de lipídios. A aplicação de Mz proporcionou o aumento da atividade da SOD, sendo assim também ocorreu o incremento POD e da CAT, consequentemente reduzindo o peróxido de hidrogênio em folhas de feijoeiro, o que manteve baixo os níveis de peroxidação lipídica. Além disso, os níveis de prolina caíram significativamente após a aplicação de Mz. Quando se utilizou o Mz associado com o ácido salicílico, se observou que ocorreu o aumento da atividade da SOD, atingindo o seu pico para a dose de 200 mg ha-1. Além disso o ácido salicílico promoveu o incremento de prolina quando associado ao Mz, em relação ao tratamento com Mz isolado. Nesta situação a prolina foi reponsável pela eliminação de H2O2, mantendo os níveis das enzimas SOD e CAT reduzidos. Com os baixos níveis de peróxido de hidrogênio a peroxidação lipídica reduziu significativamente. As aplicações de Mz refletiram na maior massa seca de vagens e implicando na maior produtividade do feijoeiro, ao passo que o ácido salicílico proporcionou o aumento de produtividade, porém dependente da dose utilizada que apresentou o melhor resultado na dose de 200 mg ha-1. / The use of multisite fungicides, such as mancozeb (Mz), has grown in the last few seasons in order to avoid fungal resistance to specific-site fungicides, in addition to increasing its efficiency in disease control. However, in addition to the fungicidal effect, in areas that have received the application of these products, physiological effects have been observed in the plants, ranging from the reduction of the stress level to the increase in chlorophyll content, which has an impact on productivity. Associated with this, salicylic acid also becomes an important strategy, since this hormone promotes reduction of stress level, besides acting in the induction of resistance against diseases. However, knowledge about physiological effects is well limited on what isolated and associated products can cause on the common bean crop. Therefore, the objective of this work was to evaluate the effect of Mz and salicylic acid applications on physiological characteristics, production components and productivity of the common bean crop. The experiment was carried out in a commercial area of irrigated common bean production in \'Patos de Minas\'-MG between April and August 2017. A randomized complete block design was used in a 5 x 2 factorial scheme consisting of five doses of salicylic acid (0, 50, 100, 200 and 400 mg ha-1) with or without the application of Mz at a dose of 400 g ha-1 and five replications. For each treatment, three applications were performed, being these at 22, 37 and 52 days after sowing. Nitrate reductase activity, urease, antioxidant enzymes (SOD, POD and CAT), proline content, hydrogen peroxide and lipid peroxidation were evaluated. The application of Mz provided the increase of the SOD activity, thus also the POD and CAT increase, consequently reducing the hydrogen peroxide in leaves of common bean, which kept the levels of lipid peroxidation low. In addition, proline levels dropped significantly after application of Mz. When Mz associated with salicylic acid was used, it was observed that the increase in SOD activity occurred, reaching its peak at the 200 mg ha-1 dose. In addition, salicylic acid promoted the increase of proline when associated with Mz, in relation to the treatment with Mz alone. In this situation proline was responsible for the elimination of H2O2, keeping the levels of the SOD and CAT enzymes reduced. With the low levels of hydrogen peroxide the lipid peroxidation reduced significantly. The Mz applications reflected in the higher dry mass of pods and implying a higher yield of common bean, while salicylic acid provided the increase in productivity, however, depending on the dose used, which presented the best result at 200 mg ha-1.

Os objetivos deste trabalho foram detectar e identificar os fungos presentes em amostras de sementes de ipê-amarelo (Tabebuia serratifolia), ipê-roxo (Tabebuia impetiginosa), aroeira-pimenteira (Schinus terebinthifolius) e aroeira-salsa (Schinus molle), coletadas em diferentes localidades (Piracicaba-SP, Mogi-Guaçu-SP, Rio das Pedras-SP, Ijaci-MG, Itumirim-MG e Lavras-MG) uma parte das amostras foram submetidas à assepsia e a outra não; verificar o efeito desses fungos na germinação dessas sementes; avaliar a transmissão de fungos das sementes para as plântulas; avaliar a patogenicidade de Pestalotiopsis sp. em mudas de aroeira-pimenteira e comparar o efeito de diferentes fungicidas (captam, carbendazim+tiram e carboxim+tiram) na incidência dos fungos e na germinação. Constatou-se que os fungos mais frequentes e em maior incidência encontrados associados às sementes, tanto de ipê-amarelo quanto de ipê-roxo, foram Cladosporium sp., Alternaria alternata, Epicoccum sp., Phoma sp., Geotrichum sp., Penicillium sp., Trichothecium sp., Phomopsis sp., Drechslera sp., Aspergillus spp., Curvularia sp. e Fusarium spp. De maneira geral, a assepsia proporcionou uma redução drástica de todos os fungos, em ambas espécies. Não houve diferença significativa na porcentagem de plântulas normais, entre as amostras, porém o tratamento com hipoclorito de sódio, reduziu a germinação em 64%. Na transmissão observou-se, em média, 17% e 10% de plântulas com sintomas, nas amostras sem assepsia e com assepsia, respectivamente. Os fungos mais freqüentes transmitidos pelas sementes de ipê foram: Alternaria alternata., Fusarium spp., Aspergillus spp., Phoma sp. e Phomopsis sp. Em relação às espécies de aroeira, os fungos quantificados foram Cladosporium sp., Alternaria alternata, Aspergillus spp., Pestalotiopsis sp., Penicillium sp., Fusarium spp., Epicoccum sp., Nigrospora sp., Curvularia sp., Drechslera sp., Trichoderma sp., Myrothecium sp. e Phoma sp. A assepsia reduziu ou manteve a incidência dos fungos, exceto para Pestalotiopsis sp. e Aspergillus spp. que aumentaram em algumas amostras de sementes de aroeira-pimenteira. Não houve diferença estatística da germinação das com e sem assepsia e entre amostras; para aroeira-salsa não verificou-se germinação. Foi confirmada a transmissão, principalmente dos fungos Cladosporium sp., Aspergillus spp. e Pestalotiopsis sp. Em mudas de aroeira-pimenteira verificou-se a patogenicidade de Pestalotiopsis sp. No tratamento de sementes com fungicidas, todos, de uma maneira geral, mostram resultados satisfatórios no controle de todos os fungos detectados, tanto para ipê quanto para aroeira. Captam apresentou resultado satisfatório principalmente em sementes de ipê, onde não foi verificado efeito fitotóxico em relação aos demais fungicidas comparados. Porém, o uso de copolímero de poliéter e silicone, um espalhante adesivo utilizado para facilitar a distribuição do produto, interferiu de forma negativa nos resultados de germinação de sementes de ipê; para aroeira não foi verificado este efeito. Torna-se necessário comparar diferentes doses e produtos para não só controlar fungos associados às sementes mas evitar problemas durante a germinação das mesmas e garantir a produção de mudas sadias e vigorosas em viveiros. / The objectives of this work were to detect and to identify the fungi present in samples of native forest seeds of ipê-amarelo (Tabebuia serratifolia), ipê-roxo (Tabebuia impetiginosa), aroeira-pimenteira (Schinus terebinthifolius) and aroeira-salsa (Schinus molle), collected at different places (Piracicaba-SP, Mogí-Guaçu-SP, Rio das Pedras-SP, Ijaci-MG, Itunirim-MG and Lavras-MG), submiting some parts of the asepsis and the other no; to verify the effect of these fungi in the germination of these seeds; to evaluate the transmission of seed fungi associated to seedlings; to evaluate the patogenicity of Pestalotiopsis sp. in aroeira-pimenteira seedlings and to compare the effect of different fungicides (captan, carbendaxim+tiram and carboxim+tiram) in the fungi incidence and germination. The results showed that the most incident and frequent fungi found associated to the seeds of ipê-amarelo and ipê-roxo were Cladosporium sp., Alternaria alternata, Epicoccum sp., Phoma sp., Geotrichum sp., Penicillium sp., Trichothecium sp., Phomopsis sp., Drechslera sp., Aspergillus spp., Curvularia sp. and Fusarium spp. In general, the asepsis provided a drastic reduction of all fungi, in both species. There was no significantive difference in the normal seedling percentage, independent of the sample, however the treatment with sodium hipoclorite, in the asepsis, reduced the germination in 64%. In the transmission was observed, on average, 17% and 10% of seedlings with symptoms, in the samples without asepsis and with asepsis, respectively. The most frequent fungi transmitted by ipê seeds were: Alternaria alternata., Fusarium spp., Aspergillus spp., Phoma sp. e Phomopsis sp.. In relation to aroeira species, the quantified fungi were: Cladosporium sp., Alternaria alternata, Aspergillus spp., Pestalotiopsis sp., Penicillium sp., Fusarium spp., Epicoccum sp., Nigrospora sp., Curvularia sp., Drechslera sp., Trichoderma sp., Myrothecium sp. e Phoma sp.. The The asepsis reduced or maintained the fungi incidence, except for Pestalotiopsis sp. and Aspergillus spp. That increased in some aroeira-pimenteira seeds. The germination results showed that there was no statistical difference in relation to seeds with and without asepsis and among samples. The transmission was confirmed, mainly of the fungi Cladosporium sp., Aspergillus spp. e Pestalotiopsis sp.. In aroeira-pimenteira seedlings the patogenicity of Pestalotiospsis sp. was verified. In the seeds treatment with fungicides, all, in a general way, showed satisfactory results in the control of all detected fungi, as much for ipê as for aroeira. Captan presented satisfactory result principally in ipê seeds, where fitotoxic effect had not been verified in relation to the other compared fungicides. However, the use of copolímero of polyeter and silicon, an adhesive dispersed, used to facilitate the product distribution, negatively interfered on the germination results; for aroeira this effect had not been verified in the germination. To compare different doses, products not only to control fungi associated to the seeds but to avoid problems during the germination of the same ones and to guarantee the production of healthy seedlings and vigorous in nurseries becomes necessary.

O aumento do consumo de agrotóxicos, em geral nos países em desenvolvimento cuja base econômica é a agricultura, como no Brasil (primeiro no ranking mundial), tem sido relacionado à expansão da degradação dos ecossistemas e à perda da biodiversidade, principalmente das espécies não alvo, incluindo ainda os riscos inerentes à saúde humana. Além disso, a ausência de políticas e programas mais efetivos relacionados à regulação e uso de agrotóxicos no Brasil, bem como o estabelecimento de normas restritivas baseadas em resultados gerados em países de clima temperado, tornam-se cada vez mais preocupantes, pois podem subestimar os reais riscos da contaminação ambiental por agrotóxicos em ecossistemas tropicais. Nesse contexto, o presente estudo teve como objetivo avaliar os efeitos diretos e indiretos do agrotóxico pyrimethanil, um fungicida utilizado em diversas culturas de climas temperado e tropical, sobre a comunidade aquática tropical, considerando experimentos laboratoriais e o uso de modelos ecossistêmicos para avaliação destes efeitos. Para atender aos objetivos propostos foram realizados testes de toxicidade com as espécies Pseudokirchneriella subcapitata (alga), Ceriodaphnia silvestrii e Daphnia similis (zooplâncton) e Danio rerio (peixes), além de um experimento em mesocosmos, monitorado em longo prazo (durante 15 dias antes e 366 dias após a contaminação), avaliando as variações físicas e químicas, bem como as respostas das comunidades zooplanctônicas e fitoplanctônicas frente à contaminação por pyrimethanil, em uma concentração média de 1,36 mg.L-1. Nos testes de toxicidade, as CEs obtidas foram de 0,67 mg.L-1 (crônica) e 3,26 mg.L-1 (aguda) para C. silvestrii; de 3,65 mg.L-1 (agudo) para D. similis; de 9,65 mg.L-1 para P. subcapitata e para D. rerio os valores obtidos para imobilidade foram de 10,45 mg.L-1 (96h) e 13,77 mg.L-1(48h) e para a mortalidade foram de 27,45 mg.L-1 (96h) e 32,17 mg.L-1 (48h). Pelos resultados dos experimentos em mesocosmos verificaram-se efeitos negativos diretos sobre o zooplâncton (redução da densidade e alteração na composição de espécies) e indiretos sobre o fitoplâncton, decorrentes da diminuição da pressão de pastagem, além do desaparecimento de algumas espécies mais sensíveis. Os efeitos tóxicos observados foram além do período em que foi observada a degradação completa do pyrimethanil, demonstrando o alto impacto do agrotóxico em ecossistemas aquáticos e a complexidade da dinâmica desse tipo de contaminação em ecossistemas naturais. O design experimental revelou-se adequado, gerando informações importantes sobre os impactos dos agrotóxicos nos ecossistemas aquáticos. A partir dos resultados obtidos sugere-se que a utilização de espécies nativas de regiões tropicais seja ampliada nos testes de toxicidade laboratoriais, sendo fundamental que os mesmos sejam complementados com os experimentos em modelos ecossistêmicos, o que permite uma avaliação dos efeitos diretos e indiretos dos contaminantes em nível de população e comunidade, bem como uma melhor compreensão destes efeitos nas funções e serviços ecossistêmicos. / The increase in the consumption of pesticides, mainly in developing countries whose economic base is agriculture, as Brazil (first in the world ranking), has been associated to the expansion of ecosystem degradation and biodiversity losses, especially of non-target species, also including the risks to human health. In addition, the absence of policies and effective programs related to the regulation and use of pesticides in Brazil, and the establishment of restrictive standards based on results generated in temperate countries, make it becomes increasingly worrisome because it may underestimate the real risks of environmental contamination by pesticides in tropical ecosystems. In this context, the present study aim to evaluate the direct and indirect effects of the pesticide pyrimethanil, a fungicide used in many temperate and tropical crops, upon a tropical aquatic community, considering laboratory experiments and the use of ecosystem models to evaluate these effects. To achieve the proposed objectives toxicity tests were performed with the species Pseudokirchneriella subcapitata (algae), Ceriodaphnia silvestrii and Daphnia similis (zooplankton) and Danio rerio (fish), as well as an experiment in mesocosms, monitored for a long-term (15 days before and 366 days after infection), evaluating the physical and chemical variations, and also the responses of phytoplankton and zooplankton communities across the pyrimethanil contamination in an average concentration of 1.36 mg.L-1. In toxicity tests, the ECs obtained were 0.67 mg.L-1 (chronic) and 3.26 mg L-1 (acute) for C. silvestrii; 3,65 mg.L-1 (acute) for D. similis; 9.65 mg.L-1 for P. subcapitata; and the values obtained for D. rerio immobility were 10.45 mg.L-1 (96h) and 13.77 mg.L-1 (48h) and mortality were 27.45 mg.L-1 (96h) and 32.17 mg.L-1 (48h). The results of the experiments in mesocosms showed direct negative effects on zooplankton (reduced density and change in species composition) and indirect effects on phytoplankton, resulted from the decrease in grazing pressure in addition to the exclusion of some sensitive species. Even after the complete degradation of pyrimethanil, toxic effects were observed, demonstrating the high impact of this pesticide on aquatic ecosystems and the complexity of the dynamics of this type of contamination in natural ecosystems. The design of the experiment revealed to be appropriate for the study, generating important information about the impacts of pesticides on aquatic ecosystems. From the results, it is suggested an increase in the use of native tropical species in the toxicity laboratory tests. Also, this process should be complemented with experiments in ecosystem models, allowing the evaluation of the direct and indirect effects of contaminants in the population and in community level as well as a better understanding of these effects on ecosystem functions and services.

Made available in DSpace on 2014-08-20T13:44:37Z (GMT). No. of bitstreams: 1 dissertacao_clarissa_da_silva.pdf: 333325 bytes, checksum: 487ed9f3127ccb664dcea12fed6e7838 (MD5) Previous issue date: 2007-03-15 / This work was conducted to evaluate the quality of rice seeds treated whit fungicide in different moisture contents during of stored period. Were used two rice seeds lots, cv EL PASO 144, produced in Pelotas/RS during 04/05. For the formularization of fungicide, were additioned 10mL (1%), 20mL (2%) e 30mL (3%) of water in mixed with fungicide Carboxin/Thiram (300mL/ 100Kg of seeds). Same water percentages, without addition of the fungicide, besides one control, had constituted the seven treatments. The seeds were stored in hermetic recipient during eight months period. The quality of seeds was submitted to moisture determination, standard germination, vigor and health. Biochemical evaluation of the seeds was obtained by the technique of electrophoresis of enzymatic systems, being evaluated the activity of the enzymes Acid Phosphates, Alcohol Dehydrogenize, Glutamate Oxalacetate Transaminase, Esterase and a- amylase. The statistic model was of randomizes completely blocks with three replications. The benefic effect of fungicide on physiological quality are evident immediately after the treatment of seeds. The results indicate the germination and vigor reduction of treated seeds increase after 60 days of storage. The fungicide used in treatments of seeds is efficient in reduction of the fungi associates on the rice seeds. Wasn t possible to identify the effect of fungicide in the expression of enzymes of viable seeds. / Este trabalho foi conduzido com objetivo avaliar a qualidade de sementes de arroz tratadas com fungicida em diferentes graus de umidade durante o período de armazenamento. Foram utilizados dois lotes de sementes da cultivar EL PASO 144, de diferentes graus de umidade e sanidade, produzidos na região de Pelotas, RS, na safra 04/05. Para a formulação da calda fungicida foram adicionados 10mL(1%), 20mL (2%), e 30mL (3%) de água em mistura com fungicida Carboxin/Thiram (300mL/100Kg de sementes). Mesmas percentagens de água, sem adição do fungicida, além de uma testemunha, constituíram os sete tratamentos. Após, foram embaladas em recipientes herméticos, durante o período de oito meses de armazenamento. A qualidade das sementes foi monitorada através do teste para determinação do grau de umidade, germinação, vigor e sanidade. Avaliações bioquímicas das sementes, através da técnica de eletroforese de sistemas enzimáticos, determinou-se a atividade das enzimas Fosfatase Ácida, Álcool Desidrogenase, Glutamato Oxalacetato Transaminase, Esterase e Alfa -amilase. O modelo estatístico utilizado foi o completamente casualizado, com três repetições. Os efeitos benéficos do tratamento fungicida sobre a qualidade fisiológica são evidentes logo após o tratamento das sementes. Os resultados obtidos demonstram decréscimo na porcentagem de germinação e vigor das sementes tratadas, intensificado a partir do 60º dia de armazenamento. O fungicida utilizado no tratamento de sementes é eficiente na redução da incidência de fungos associados à semente. Não foi possível detectar o efeito do fungicida na expressão das enzimas das sementes viáveis.

The global human population is expected to exceed 9 billion individuals by 2050, putting greater strain on the natural resources needed to sustain such a population. To feed this many people, some expect agricultural production will have to double and agrochemical use will have to increase anywhere from two- to five-fold relative to the turn of the century. Although industrial agriculture has provided many benefits to society, it has caused declines in biodiversity, both directly (e.g., through conversion of habitat) and indirectly (e.g., through contamination of adjacent natural habitats). Agricultural activity has also been linked to increased prevalence and intensity of trematode infections in wildlife and humans - directly by increasing available aquatic habitat for the snail intermediate hosts of trematode parasites and indirectly by altering the biological composition of aquatic habitats in ways that increase snail density. While the effects of single agrochemical contaminants on aquatic communities and trematode disease risk have been examined, agrochemical pollution typically occurs as mixtures of multiple chemical types in surface waters and the effects of mixtures on aquatic communities have received less attention. Moreover, given the high number of chemicals approved for agricultural use, the number of potential combinations of agrochemicals renders testing all possible combinations implausible. Thus, there is a critical need to develop better risk assessment tools in the face of this complexity. I developed and tested a theoretical framework that posits that the net effects of agrochemical mixtures on aquatic communities can be predicted by integrating knowledge of each functional group's 1) sensitivity to the chemicals (direct effects), 2) reproductive rates (recovery rates), 3) interaction strength with other functional groups (indirect effects), and 4) links to ecosystem properties. I conducted a freshwater mesocosm experiment to quantify community- and ecosystem-level responses to pairwise mixtures of four major agrochemical types (fertilizer, herbicide, insecticide, and fungicide) and single chemical treatments. The responses of biodiversity and ecosystem properties to agrochemicals alone and in mixtures were indeed predictable. Moreover, these results show that community ecology theory holds promise for predicting the effects of contaminant mixtures and offer recommendations on which types of agrochemicals to apply together and separately to reduce their impacts on aquatic ecosystems. I extended this framework to test if the direct effects of pesticides can be predicted by chemical class and/or mode of action. I performed standard toxicity trials on two invertebrate predators of snails (crayfish and giant water bugs) exposed to six insecticides belonging to two chemical classes (organophosphates and pyrethroids) to determine if environmental risk can be generalized to either insecticide class or insecticide exposure. Survival analyses demonstrated that insecticide class accounted for 55.7% and 91.1% of explained variance in crayfish and water bug survival, respectively. Simulated environmental exposures using US EPA software suggested that organophosphate insecticides present relatively low risk (as defined by the US EPA) to both crayfish and water bugs, while pyrethroid insecticides present consistently high risk to crayfish but not to water bugs, where only λ-cyhalothrin produced consistently high-risk exposure scenarios. Thus, risk to non-target organisms is well predicted by pesticide class. Furthermore, identifying insecticides that pose low risk to aquatic macroarthropods might help meet increased demands for food while mitigating against potential negative effects on ecosystem functions. Because evidence from field data and manipulated experiments demonstrated both top-down and bottom-up effects of agrochemical pollution that increased snail densities and trematode infections in wildlife, I conducted an additional agrochemical mixture experiment with freshwater communities containing the snail hosts of schistosomiasis, which has also been linked to agriculture. As expected, top-down and bottom-up effects of insecticide, herbicide, and fertilizer exposure indirectly increased snail densities, individually and as mixtures. Agrochemical exposure and snail density together accounted for 88% of the variation in the density of infected snails. Thus, agrochemical pollution has great potential to increase human exposure to schistosome parasites, and underscores the importance of identifying low-risk alternative pesticides. A subsequent mesocosm experiment with the same six insecticides used previously in laboratory trials confirmed that insecticide exposure indirectly mediates the densities of snail hosts that can transmit schistosomiasis through the direct effects of insecticides on crayfish mortality. Importantly, crayfish mortality in semi-natural mesocosm trials closely matched mortality from controlled laboratory trials. Thus, standard laboratory toxicity tests can be a useful tool for identifying alternative insecticides that might pose lower environmental risks to important predators that regulate snail densities. Identifying practices or agrochemicals that minimize this risk is critical to sustainably improving human health in schistosome-endemic regions. The theoretical framework presented here demonstrates the feasibility of predicting the effects of contaminant mixtures and highlights consistent effects of major agrochemical types (e.g. fertilizers, insecticides, etc.) on freshwater aquatic community composition. Furthermore, the strong top-down effects of invertebrate snail predators highlight that managing for high snail predator densities in might be a particularly effective strategy for reducing the burden of schistosomiasis in tropical countries.

Azoxystrobin (AZY), a systemic broad-spectrum fungicide, is applied on crops to control soil-borne pathogenic fungi. This study aimed to determine the effects of AZY on non-target plant species and Glomus intraradices Schenck & Smith, an arbuscular mycorrhizal fungus (AMF) associated with plants' roots. We hypothesized that AZY negatively affects AMF viability; and that, if the plants were dependent on this symbiosis, AZY exerts an indirect detrimental effect on plant growth. To test this, three mycotrophic (Phalaris arundinacea L., Solidago canadense L., Geum canadense Jacq.) and one non-mycotrophic (Chenopodium album L.) native plant species were subjected to five AZY doses with or without AMF. Plants were grown for 60 days in a greenhouse, in individual pots, (4 plants X 2 AMF X 5 AZY X 6 replicates), and mesocosms (1 mes. X 2 AMF X 5 AZY X 6 replicates), and harvested 30 days after spraying, and dry mass was taken. Fresh root samples were used for microscopic assessment of AMF colonization. The results from the individual pot experiment show that the effects of AZY on biomass varied across plant species. AZY led to a significant increase in shoot and root mass of P. arundinacea, and a decrease in shoot mass of AMF inoculated G. canadense. The presence of AMF resulted in a significant increase in root and shoot mass of P. arundinacea, and an increase in root mass of S. canadense and shoot mass of C. album. In the mesocosm experiment AZY did not have a significant effect on the measured parameters, although the presence of AMF significantly increased root, shoot, and total dry mass of G. canadense and P. arundinacea. Conversely, AMF significantly decreased shoot and total dry mass of S. canadense. The results suggest that both direct and indirect effects should be taken into account when assessing the impact of pesticides on non-target plant species.

La perception et l’adaptation à l’environnement sont des processus indispensables pour la survie des organismes vivants. Le champignon phytopathogène Botrytis cinerea peut ainsi percevoir différents types de signaux qu’ils soient chimiques ou physiques. La voie de signalisation de la MAPK Sak1 est impliquée dans l’adaptation au stress osmotique, oxydatif et pariétal, mais aussi dans la sporulation et le pouvoir pathogène en régulant la pénétration de la plante et le développement des nécroses. Afin d’approfondir les connaissances existantes sur la voie de Sak1, nous avons réalisé des études globales basées sur des techniques de protéomique et phosphoprotéomique. L’analyse de protéomique comparative entre la souche sauvage et les mutants de signalisation ∆bos1 et ∆sak1 a notamment mis en évidence que la MAPK Sak1 régule l’abondance de protéines impliquées dans la voie des protéines G et la voie calcique. Cette connexion avec les protéines G a été confirmée par une baisse de la concentration en AMPc chez le mutant ∆sak1. L’utilisation du fludioxonil comme signal de l’activation de la MAPK Sak1 pour l’analyse par phosphoprotéomique a mis en évidence des modifications de l’état de phosphorylation de protéines. Parmi ces protéines différentiellement phosphorylées, la présence de PKAR (sous-unité régulatrice de la protéine kinase A) et du facteur de transcription CRZ1, indiquent respectivement une action sur la voie via protéines G et la voie calcique, validant les résultats obtenus par protéomique. Le phosphoprotéome a révélé une « phosducin-like protein », PhnA. Sa caractérisation fonctionnelle montre son rôle dans l’adaptation aux stress, la sporulation et la germination, ainsi que dans le pouvoir pathogène mettant ainsi en évidence un nouveau facteur de pathogénicité chez B. cinerea. Notre étude a permis de révéler des interactions entre Sak1 et d’autres voies de signalisation non suspectées, agissant aussi bien sur la production de certains composants (régulations transcriptionnelles et traductionnelles) que sur la phosphorylation (modifications post-traductionnelles). Nos résultats constitueront la base de nouvelles recherches pour compléter nos connaissances sur ces interactions impliquant l’adaptation au stress et la pathogénie de B. cinerea. / Perception and adaptation to the environment are essential processes for the survival of living organisms. The phytopathogenic fungus Botrytis cinerea can thus perceive different types of signals, whether they are chemical or physical. The signalling pathway of the Sak1 MAPK is involved in the adaptation to osmotic, oxidative and cell wall stress, but also in sporulation and pathogenicity by regulating plant penetration and necrosis development. In order to deepen existing knowledge of the Sak1 pathway, we have carried out global studies based on proteomics and phosphoproteomics techniques. A comparative proteomics analysis between the wild type and the signalling mutants ∆bos1 and ∆sak1 showed, among others, that Sak1 regulates the abundance of proteins involved in the G-protein pathway and calcium pathway. This connection with G-proteins was confirmed by a decrease in cAMP concentration in the ∆sak1 mutant. Using fludioxonil as signal for the activation of Sak1 for a phosphoproteomic analysis revealed changes in the state of protein phosphorylation. Among these differentially phosphorylated proteins, the presence of PKAR (regulatory subunit of protein kinase A) and the transcription factor CRZ1, indicates an action on the G-protein and calcium pathway respectively, validating the results obtained by proteomics. Phosphoproteomics revealed a phosducin-like protein, PhnA. Its functional characterization reveals its role in stress adaptation, sporulation and germination, as well as in pathogenicity, thus demonstrating a new pathogenicity factor in B. cinerea. Our study revealed interactions between Sak1 and other unsuspected signalling pathways, affecting both the production of certain components (transcriptional and translational regulations) and phosphorylation (post-translational modifications). Our results will create the basis for new research questions to complement our understanding of these interactions involving adaptation to stress and pathogenesis of B. cinerea.

‘CL151’ a Clearfield long-grain rice cultivar, was released in 2008, based upon excellent agronomic characteristics, including excellent yield and moderately resistant rating for lodging. Further experience has indicated it’s susceptibility to lodging. Lodging can reduce harvest efficiency, yield, and cause grain quality loss. The purpose of this research was to evaluate multiple management practices such as nitrogen rates and timings, seeding rate, trinexapac-ethyl application, potassium, and fungicide application to mitigate lodging for CL151. The results of this research indicate that N rate and application timing largely influence lodging incidence and grain yield. Fungicide application decreased, but did not eliminate lodging, while potassium application did not impact lodging or grain yield. Seeding rate also has a significant impact on grain yield and lodging incidence. Trinexapac-ethyl also decreased lodging incidence, but negatively impacted grain yield which was most evident when applied at 48 g ha-1 and applied at PD +14d growth stage.