Cette étude visait à suivre l’abondance relative d’une combinaison de quatre souches de Fusarium graminearum au cours du processus d’inoculation artificielle de parcelles d’orge. Trois amplicons présentant des polymorphismes (SSR, SNP) spécifiques ont été examinés par pyroséquençage 454 sur des échantillons récoltés à tous les stades (souches pures, mélange des souches, inoculum frais, inoculum au champ, grains d’orges infectés). Seuls les marqueurs SNP ont permis d’obtenir une signature unique à chaque souche et ont permis d’estimer l'abondance des souches dans chaque échantillon. Cette analyse a révélé qu’il était difficile d’obtenir des mélanges parfaitement égaux des souches et que les souches se développaient de manière inégale sur les grains de maïs servant de support à l’inoculum. Au champ, ces différences se sont largement maintenues, tandis que des profils d’abondance distinctifs sont apparus sur les trois sites d’essai chez les grains d’orge infectés, mais sans différences significatives entre les cultivars d’orge. / The aim of this study was to track the evolution of a combination of four Fusarium graminearum strains within a mixed inoculum and the ensuing stages of artificial inoculation on barley cultivars. Three amplicons containing specific polymorphisms (SSR, SNP) were examined by 454 pyrosequencing on samples collected at all stages of the process (pure strains, strain mixture, primary inoculum, inoculum in the field and infected barley kernels). Only SNP markers provided unique and accurate signatures for each strain. This analysis revealed that it was difficult to obtain perfectly equal mixtures of strains and that these strains differ in their development on corn kernels used as a growth medium for the inoculum. In the field, these differences remained, while distinctive abundance profiles were detected in infected barley kernels samples at the three nurseries; however, no significant differences between barley cultivars were observed.

Mycotoxins are toxic secondary metabolites produced by fungi that are a threat to the health of humans and domestic animals. The most important mycotoxin in the U.S. is deoxynivalenol (DON), which causes symptoms such as vomiting, feed refusal, and weight loss in farm animals. The fungus Fusarium graminearum produces DON in staple crops such as wheat, barley, and corn. It is estimated that the economic losses associated with DON contamination alone exceed $650 million per year in the U.S. New strategies are needed to mitigate DON and improve food safety in the U.S. The overall goal of my research is to discover and employ microorganisms and enzymes to detoxify DON. The specific objectives are to: (1) discover and characterize microorganisms that detoxify DON, (2) use a cell free protein synthesis (CFPS) system to study enzymes that modify DON, (3) engineer yeast to detoxify DON with a metabolic engineering strategy, and (4) deliver a high school unit to teach high school students about mycotoxins in food. In Objective 1, two mixed cultures were identified from environmental samples that converted DON into the less toxic 3-keto-deoxynivalenol (3-keto-DON). In Objective 2, a CFPS system was used to express three known acetyltransferase genes to convert DON to 3-acetyl-DON (3-A-DON). In Objective 3, we identified a potential DON transporter from a library of randomly amplified fragments from the genomes of mixed cultures of microbes isolated from the environment. In Objective 4, we developed and delivered a unique high school unit to educate high school students about potential mycotoxins in food and feed products. The work presented here represents new and improved methods for mitigating mycotoxin contamination in the United States. / Ph. D.

Made available in DSpace on 2017-07-10T17:37:37Z (GMT). No. of bitstreams: 1 Josiel_Pereira_Nascimento.pdf: 909698 bytes, checksum: 3cb3ef0ef8bdf27d1c63a9fb56af8240 (MD5) Previous issue date: 2009-08-29 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / The non tillage planting and the cultivation of safrinha (short season) corn are factors that may provide alterations in the microclimate and in the biology of agro ecosystem, increasing the populations of fungus like Gibberella zeae, which is the etiologic agent of one of the maize ear rots. This work had the objective to identify inoculation methods that can be useful in the election and in the characterization of genotypes tolerant to G. zeae. The experiment was conducted in Toledo/PR during the 2008 safrinha season in a randomized complete block design with five replications, on a factorial scheme where hybrids were the first factor (2B710; 30F45; DKB 214; 30F98; DKB 390; GAC 9020; 30F80; 30R50 and 30K73) and the inoculation methods were the second factor. The hybrids had been grouped as susceptible (2B710; 30F45; DKB 214; 30K73), tolerant (DKB 390; GAC 9020; 30F80; 30R50) and moderately susceptible to giberela (30F98). The 12 evaluated methods consisted of simulating the fungus natural infection on the silk or on the damage caused by insects in the lateral of the ears, and also varying by the absence of covering on the ear (humid chamber), by covering the ear for 24 hours and by covering the ear for 48. The methods with inoculation of 2 mL in the lateral of the ear without covering (3), with covering for 24 hours (7) and with covering for 48 hours (11) had shown efficiency in the characterization of tolerant and susceptible hybrids and could be used on a large scale in the identification and selection of maize varieties with tolerance to ear rot caused by G. zeae. As a result, the seed industry can provide superior materials and the farmers will reduce their losses due to kernel quality. Moreover, the whole society will consume better maize products, with less mycotoxins / O plantio direto e o cultivo de milho safrinha são fatores que podem proporcionar alterações no microclima e na biologia do agroecossistema, aumentando as populações de fungos como Gibberella zeae, agente etiológico da doença denominada giberela ou podridão-vermelha-da-espiga. Esse trabalho teve como objetivo identificar métodos de inoculação que auxiliassem na seleção e na caracterização de genótipos tolerantes à G. zeae. O experimento foi conduzido no município de Toledo/PR durante a safrinha de 2008. O delineamento experimental foi em blocos ao acaso com cinco repetições, em esquema fatorial tendo como primeiro fator os híbridos (2B710; 30F45; DKB 214; 30F98; DKB 390; AG 9020; 30F80; 30R50 e 30K73) e como segundo fator os métodos de inoculação. Os híbridos foram agrupados como suscetíveis (2B710; 30F45; DKB 214; 30K73), tolerantes (DKB 390; AG 9020; 30F80; 30R50) e moderadamente suscetível a giberela (30F98). Os 12 métodos avaliados consistiam em simular a infecção natural do fungo pelo estigma ou por danos causados por insetos nas laterais das espigas, variando ainda em ausência de cobertura (câmara úmida), cobertura por 24 horas e por 48 horas. Os métodos com inoculação de 2 mL na lateral da espiga sem cobertura (3), com cobertura por 24 horas (7) e com cobertura por 48 horas (11) se mostraram eficientes na caracterização de híbridos tolerantes e suscetíveis e poderão ser empregados em larga escala na identificação e seleção de variedades de milho tolerantes à doença. Mais do que isso, a sociedade poderá consumir produtos de milho com qualidade superior, com menos micotoxinas

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Submitted by Claudia Rocha (claudia.rocha@udesc.br) on 2018-02-28T16:34:26Z No. of bitstreams: 1 PGPV16MA215.pdf: 500238 bytes, checksum: d01d8d01d1ca49176f3a0bdc5c42b7e5 (MD5) / Made available in DSpace on 2018-02-28T16:34:26Z (GMT). No. of bitstreams: 1 PGPV16MA215.pdf: 500238 bytes, checksum: d01d8d01d1ca49176f3a0bdc5c42b7e5 (MD5) Previous issue date: 2016-08-19 / PROMOP / Gibberella ear rot (GER) caused by Fusarium graminearum reduces the yield and quality of maize grain, interfering in the presence of damaged grains (DG). The disease control is hampered by unavailability of resistance hybrids and effectiveness of cultural practices to reduce fungal inoculum sources. The objective of this study was to evaluate the effect of preventative and curative fungicide application during maize silking on the GER control and the effect on grain yield (GY) and incidence of DG. The study was conducted in experimental area of Santa Catarina State University, in Lages, Brazil in two sowing dates in 2014/15 growing seasons, using the hybrid P32R48YH. Two F. graminearum isolates, one from Minas Gerais (MEMR0004) and other from Paraná state (MEMR0005) were inoculated in the maize ears five days after silking. Eight fungicides belonging to the chemical group of triazoles, triazolinthione, strobilurins, and benzimidazole, individually or in combination were tested. Maize plants inoculated with fungus without fungicide applicaton were used as experiment control. An application with different fungicides was carried out on tassels with pressurized spray with flow rate of 200 L ha-1, 48 hours before inoculation (preventive action) or 48 hours after inoculation (curative action). There was homoscedasticity between sowing dates and the data were analyzed together. Differences statistics were not found between treatments to GER severity and DG. Preventative application of fungicides (mean values) to MG isolate reduce GER severity and increased GY differing from curative application. Isolated PR showed higher percentages of DG differing from the isolated MG. In PR isolated curative application of fungicides affected the GY with a significant difference between treatments. The fungicides trifloxystrobin + prothioconazol and azoxystrobin + cyproconazol + carbendazim showed stability in control of GER keeping the lower severity to 46%. These fungicidal beyond carbendazim used alone reduced the DG incidence. The tebuconazol + trifloxystrobin and azoxystrobin + cyproconazol + carbendazim in preventive application maintained GY higher than control, in curative were tebuconazol, trifloxystrobin + prothioconazol and methyl thiophanate. The fungicides application on corn silking is a strategy control that can be used in the integrated management of GER to maintain hybrid potential production and grain quality due to reduction in the percentage of DG / A podridão da espiga de giberela (PEG), causada pelo fungo Fusarium graminearum, reduz o rendimento e a qualidade de grãos de milho, interferindo na presença de grãos avariados (GA). O controle da doença é dificultado pela indisponibilidade de híbridos resistentes e pela baixa eficácia de práticas culturais em reduzir as fontes de inóculo do fungo. O objetivo deste trabalho foi avaliar o efeito da aplicação preventiva e curativa de fungicidas no espigamento do milho no controle da PEG e na influência sobre rendimento de grãos (RG) e incidência de GA. O estudo foi conduzido na área experimental da Universidade do Estado de Santa Catarina, no município de Lages, Brasil em duas épocas de semeadura na safra 2014/15, utilizando o híbrido P32R48YH. Dois isolados de F. graminearum um proveniente de Minas Gerais (MEMR0004) e outro do Paraná (MEMR0005) foram inoculados nas espigas cinco dias após emissão dos estilos-estigmas. Oito fungicidas pertencentes aos grupos químicos dos triazois, triazolintiona, estrobilurinas e benzimidazóis isolados ou em mistura foram testados. Plantas de milho inoculadas sem aplicação de fungicidas foram utilizadas como controle experimental. Uma aplicação dos diferentes fungicidas foi realizada sobre os pendões com auxílio de pulverizador pressurizado com vazão de 200 L ha-1, 48 horas antes da inoculação (ação preventiva) e 48 horas depois da inoculação (ação curativa). Houve homocedasticidade entre as épocas de semeadura sendo os dados analisado de forma conjunta. Não houve diferença estatística na severidade da PEG e GA entre os tratamentos. Aplicação preventiva de fungicidas (valores médios) no isolado MG reduziu a severidade da PEG e aumentou o RG diferindo da aplicação curativa. O isolado PR apresentou maiores percentuais de GA diferindo do isolado MG. No isolado PR a aplicação curativa de fungicidas afetou o RG havendo diferença significativa entre os tratamentos. Os fungicidas trifloxistrobina + protioconazole e azoxistrobina + ciproconazole + carbendazim apresentaram estabilidade no controle da PEG mantendo a severidade inferior a 46%. Esses fungicida além do carbendazim utilizado isoladamente reduziram a incidência de GA. Os fungicidas tebuconazole + trifloxistrobina e azoxistrobina + ciproconazole + carbendazim aplicados de forma preventiva mantiveram o RG superior ao controle, em aplicações curativas foram tebuconazol, trifloxistrobina + protioconazole e tiofanato metílico. A aplicação de fungicidas no espigamento do milho é uma estratégia de controle que pode ser utilizada no manejo integrado da podridão da espiga de giberela com a finalidade de manter o potencial produtivo do hibrido e a qualidade de grãos devido a redução do percentual de GA

La fusariose des épis est l’une des principales maladies des céréales, majoritairement causée par le champignon pathogène et toxinogène, Fusarium graminearum (Fg). Lors son développement in planta, le champignon produit des mycotoxines dommageables pour la santé humaine et animale, dont le déoxynivalénol (DON). De nombreux loci à effet quantitatif sur la résistance à Fg ont été identifiés chez le blé tendre. Certains d’entre eux ont été corrélés à la capacité à détoxifier le DON, en particulier par glucosylation sous l’action d’UDP-glucosyltransférases (UGT). Une UGT d’orge impliquée dans la conjugaison du DON a été identifiée en système hétérologue. Brachypodium distachyon (Bd) a récemment émergé comme modèle d’étude pour les céréales. Ce travail à l’aide d’approches transcriptomique et métabolomique a mis en évidence que lors de l’interaction avec Fg, Bd met en place des réponses macroscopiques, moléculaires et métaboliques similaires à celles connues chez le blé et l’orge. La recherche d’UGTs candidates capables de conjuguer le DON en DON-3-O-glucoside (D3G) chez Bd a permis l’identification d’un candidat. L’analyse fonctionnelle du gène correspondant a été conduite par des approches de mutagenèse et de surexpression. Ceci a montré une sensibilité accrue des lignées mutantes à la toxine et à l’agent pathogène. A l’inverse les lignées surexpresseurs ont montré une tolérance et résistance quantitative à la toxine et l’agent pathogène. Ces résultats ont été corrélés par la détection in planta de DON et D3G, dans des proportions variables selon les lignées. Ces résultats démontrent le rôle majeur que joue la glucosylation du DON dans l’établissement de la résistance observée chez Bd en réponse à Fg. / Fusarium head blight is a major cereal disease, mostly caused by the pathogenic and toxin-producing fungus, Fusarium graminearum (Fg). During its development in planta, the fungus produces mycotoxins harmful to human and animal health, including deoxynivalenol (DON). Many quantitative trait loci exhibiting an effect on resistance to Fg have been identified in wheat. Some of them were correlated with the ability to detoxify DON, particularly by glucosylation by UDP-glycosyltransferases (UGT). A barley UGT involved in the conjugation of DON was identified in a heterologous system. Brachypodium distachyon (Bd) has recently emerged as a model species for cereals. Using transcriptomic and metabolomic approaches, we show that when interacting with Fg, Bd implements macroscopic, molecular and metabolic responses similar to those known in wheat and barley. The search for UGT candidates able to conjugate DON into DON-3-O-glucoside (D3G) in Bd resulted in the identification of the Bradi5g03300 gene. Functional analyses of this gene showed increased sensitivity of the mutant lines to the toxin and to the pathogen. Conversely the overexpressor lines showed a tolerance to the toxin and quantitative resistance to Fg. These results were correlated with the detection of differential amounts of DON and D3G in the different lines. These results demonstrate the important role of DON glucosylation in the resistance establishment of Bd observed in response to Fg.

Master of Science / Department of Plant Pathology / Robert L. Bowden / John F. Leslie / Fusarium graminearum Schwabe sensu lato (teleomorph: Gibberella zeae (Schwein.) Petch), a homothallic ascomycete fungus, is the causal agent of Fusarium head blight (FHB) of wheat (Triticum aestivum L.), barley (Hordeum vulgare L.), and other small grains. FHB occurs worldwide and serious outbreaks have been reported in North America, South America, Asia, and Europe. According to the phylogenetic species concept (PSC), F. graminearum is composed of at least 15 phylogenetic lineages known as the Fusarium graminearum species complex. Although F. graminearum is homothallic, some members of different phylogenetic lineages are known to intercross in the laboratory. It has been suggested that F. graminearum sensu lato fits the biological species concept (BSC). According to the BSC, “species are groups of actually or potentially interbreeding natural populations, which are reproductively isolated from other such groups”. Previous reports of intercrossing were qualitative, so the degree of reproductive isolation, if any, is not clear. Since intrinsic reproductive isolation is the key criterion to identify species by the BSC, more detailed quantitative information is needed. Chromosome rearrangements between fungal strains may reduce fertility in sexual crosses through the production of genetically inviable recombinant progeny. As such, rearrangements can be important postzygotic reproductive barriers between species. Following methods used in Neurospora crassa, ascospore tetrads were analyzed for patterns of ascospore viability. Crosses were made with three lineage 7 (F. graminearum sensu stricto according to PSC) strains as female. Each female was a MAT1-2 knockout mutant that rendered it obligately heterothallic. Males were several members of lineages 6 (F. asiaticum according to PSC) and lineage 7. Crosses with lineage 7 males formed complete asci with 8 ascospores indicating that their genomes are isosequential with the testers. Crosses with one strain from lineage 6 with two known inversions produced asci containing 8, 6, and 4 ascospores, consistent with it not being isosequential. However, three other strains of lineage 6 appeared to be isosequential with the testers. Therefore, chromosome rearrangements did not appear to be common to strains of lineage 6 and probably do not contribute significantly to reproductive isolation of lineage 6 and lineage 7. Interlineage fertility studies with the three lineage 7 tester strains were performed to quantify interlineage fertility parameters including the total number of ascospores produced, perithecial density, and perithecium internal development scores. All lineage 7 female testers successfully crossed to all 23 male strains from lineages 1 to 9. For total ascospore production, one female tester crossed equally well with all lineages and the other two testers showed statistically significant differences for a few lineages. For perithecial density, there was a significantly lower density with all three testers when crossed with lineage 6, but the other lineages were not statistically different from lineage 7. For perithecial development, there was large variation for every lineage. Therefore, in the crosses with reduced fertility, the reduction can be attributed to a postzygotic effect since mature perithecia and asci developed. All of the tested lineages of the Fusarium graminearum species complex can produce viable progeny with F. graminearum lineage 7, which was the taxonomic type of the original species before it was split into phylogenetic species. There are a few examples of reduced fertility with two lineage 7 testers, the remaining tester crossed equally well with all lineages. Therefore members of lineages 1-9 all should be considered members of Fusarium graminearum according to the BSC. The existing female testers could be used to identify members of the F. graminearum clade by performing test crosses in the laboratory. The PSC and BSC species concepts do not agree for this group of fungi. This disagreement indicates that the F. graminearum species complex is in the early stages of speciation. The lack of intrinsic reproductive barriers supports the hypothesis that these lineages have developed in geographic isolation. As the lineages have apparently been brought together through global trade, interlineage hybrids have been reported in the field. The discrepancy between PSC and BSC will eventually be resolved by whether the lineages fuse or remain separate in nature. Even if the lineages remain separate, this study demonstrates the potential for gene flow between lineage 7 and lineages 1 through 9.

A cultura de trigo (Triticum aestivum L.) é a segunda maior do mundo e o Brasil ocupa o segundo lugar de produção na América do sul. Entretanto, a produtividade desta cultura pode ser limitada devido à ocorrência de doenças como a brusone, causada pelo fungo Pyricularia grisea e a doença denominada giberela, causada pelo fungo Fusarium graminearum Populações bacterianas associadas à rizosfera de trigo podem apresentar potencial como agentes de controle biológico de diferentes fitopatógenos. Neste contexto, esta pesquisa foi direcionada ao estudo da composição da comunidade bacteriana rizosférica do trigo e a busca por micro-organismos com potencial para o controle biológico da brusone e da giberela. Assim, para melhor compreensão das comunidades associadas ao trigo, foram realizadas coletas em duas regiões diferentes no Brasil, sendo possível a obtenção de 606 estirpes entre bactérias e actinobactérias da rizosfera do trigo e de solo de cultivo da mesma cultura. Destas, 16 apresentaram, em testes in vitro, potencial antagonista diante dos fungos fitopatogênicos Pyricularia grisea e Fusarium graminearum com diferentes porcentagens de inibição. Dez dos isolados selecionados apresentaram similaridade com a família Streptomycetaceae, porém, quatro linhagens necessitam de estudos mais detalhados, pois a similaridade foi baixa, podendo indicar uma espécie ainda não descrita; quatro linhagens demonstraram similaridade com a família Bacillaceae e dois com a família Paenibacillaceae. Na avaliação de produção de metabólitos secundários com efeito inibitório, apenas dez apresentam potencial, porém estudos mais detalhados se fazem necessários para a confirmação deste mecanismo. A análise de diversidade bacteriana demonstrou uma maior abundância do filo Actinobacteria, seguido pelo filo Proteobacteria e Acidobacteria em ambas as áreas amostradas, entretanto, o filo Acidobacteria foi o que demonstrou a maior variação entre as classes presentes nas diferentes regiões estudadas, indicando uma seleção da comunidade de acordo com a variedade do cultivar e o estádio de desenvolvimento do vegetal. A comunidade bacteriana de trigo apresenta micro-organismos com potencial para a inibição dos fungos causadores da brusone e da giberela, porém o efeito destas linhagens deve ser melhor investigado em condições de campo. A compreensão das comunidades bacterianas associadas ao trigo pode se apresentar como uma importante ferramenta para direcionar a busca por antagonistas. / Wheat (Triticum aestivum) is the second largest crop in the world and Brazil is in the second position in the ranking of production in South America. However, its productivity can be limited due to the occurrence of diseases like wheat blast, caused by the fungus Pyricularia grisea and the disease called Fusarium head blight (FHB), caused by the fungus Fusarium graminearum. Bacterial populations associated to wheat rhizosphere may have potential to act as biological control agents of different plant pathogens. In this context, this research aimed to look at wheat rhizosphere bacterial community and the pursuit of microorganisms with potential for the biological control of wheat blast and FHB. Given this, in order to study wheat bacterial communities, data collection was carried out in two different regions in Brazil, returning 606 bacterial and actinomycetes isolates from wheat rhizosphere and bulk soil. Among these,, 16 strains revealed antagonistic potential against both plant pathogens Pyricularia grisea and Fusarium graminearum, with different percentages of inhibition. Ten strains were selected out of the 16 and showed similarity with the family Streptomycetaceae, whereas four of them displayed a low similarity, requiring a deeper analysis and might indicate new species. Four isolates showed similarity with the family Bacillaceae and two with the family Paenibacillaceae. On the assessment of production of secondary metabolites with inhibitory effects, only ten strains were positive, but more detailed studies are necessary to confirm this mechanism. The analysis of bacterial diversity revealed a larger abundance of the phylum Actinobacteria, followed by the phylum Proteobacteria and Acidobacteria in both areas, however, the phylum Acidobacteria revealed more variation among its classes when both araes were compared, indicating a selection of the community according to the cultivar and the developmental stage. Wheat bacterial community presents microorganism with inhibition potential against fungi responsible for wheat blast and FHB, yet the effect of such strains should be investigated closely under field conditions. The understanding of bacterial communities associated to wheat may be seen as an important tool to help in the search for antagonists.

A cultura de trigo (Triticum aestivum L.) é a segunda maior do mundo e o Brasil ocupa o segundo lugar de produção na América do sul. Entretanto, a produtividade desta cultura pode ser limitada devido à ocorrência de doenças como a brusone, causada pelo fungo Pyricularia grisea e a doença denominada giberela, causada pelo fungo Fusarium graminearum Populações bacterianas associadas à rizosfera de trigo podem apresentar potencial como agentes de controle biológico de diferentes fitopatógenos. Neste contexto, esta pesquisa foi direcionada ao estudo da composição da comunidade bacteriana rizosférica do trigo e a busca por micro-organismos com potencial para o controle biológico da brusone e da giberela. Assim, para melhor compreensão das comunidades associadas ao trigo, foram realizadas coletas em duas regiões diferentes no Brasil, sendo possível a obtenção de 606 estirpes entre bactérias e actinobactérias da rizosfera do trigo e de solo de cultivo da mesma cultura. Destas, 16 apresentaram, em testes in vitro, potencial antagonista diante dos fungos fitopatogênicos Pyricularia grisea e Fusarium graminearum com diferentes porcentagens de inibição. Dez dos isolados selecionados apresentaram similaridade com a família Streptomycetaceae, porém, quatro linhagens necessitam de estudos mais detalhados, pois a similaridade foi baixa, podendo indicar uma espécie ainda não descrita; quatro linhagens demonstraram similaridade com a família Bacillaceae e dois com a família Paenibacillaceae. Na avaliação de produção de metabólitos secundários com efeito inibitório, apenas dez apresentam potencial, porém estudos mais detalhados se fazem necessários para a confirmação deste mecanismo. A análise de diversidade bacteriana demonstrou uma maior abundância do filo Actinobacteria, seguido pelo filo Proteobacteria e Acidobacteria em ambas as áreas amostradas, entretanto, o filo Acidobacteria foi o que demonstrou a maior variação entre as classes presentes nas diferentes regiões estudadas, indicando uma seleção da comunidade de acordo com a variedade do cultivar e o estádio de desenvolvimento do vegetal. A comunidade bacteriana de trigo apresenta micro-organismos com potencial para a inibição dos fungos causadores da brusone e da giberela, porém o efeito destas linhagens deve ser melhor investigado em condições de campo. A compreensão das comunidades bacterianas associadas ao trigo pode se apresentar como uma importante ferramenta para direcionar a busca por antagonistas. / Wheat (Triticum aestivum) is the second largest crop in the world and Brazil is in the second position in the ranking of production in South America. However, its productivity can be limited due to the occurrence of diseases like wheat blast, caused by the fungus Pyricularia grisea and the disease called Fusarium head blight (FHB), caused by the fungus Fusarium graminearum. Bacterial populations associated to wheat rhizosphere may have potential to act as biological control agents of different plant pathogens. In this context, this research aimed to look at wheat rhizosphere bacterial community and the pursuit of microorganisms with potential for the biological control of wheat blast and FHB. Given this, in order to study wheat bacterial communities, data collection was carried out in two different regions in Brazil, returning 606 bacterial and actinomycetes isolates from wheat rhizosphere and bulk soil. Among these,, 16 strains revealed antagonistic potential against both plant pathogens Pyricularia grisea and Fusarium graminearum, with different percentages of inhibition. Ten strains were selected out of the 16 and showed similarity with the family Streptomycetaceae, whereas four of them displayed a low similarity, requiring a deeper analysis and might indicate new species. Four isolates showed similarity with the family Bacillaceae and two with the family Paenibacillaceae. On the assessment of production of secondary metabolites with inhibitory effects, only ten strains were positive, but more detailed studies are necessary to confirm this mechanism. The analysis of bacterial diversity revealed a larger abundance of the phylum Actinobacteria, followed by the phylum Proteobacteria and Acidobacteria in both areas, however, the phylum Acidobacteria revealed more variation among its classes when both araes were compared, indicating a selection of the community according to the cultivar and the developmental stage. Wheat bacterial community presents microorganism with inhibition potential against fungi responsible for wheat blast and FHB, yet the effect of such strains should be investigated closely under field conditions. The understanding of bacterial communities associated to wheat may be seen as an important tool to help in the search for antagonists.

F. graminearum est l’une des espèces prédominantes en tant que pathogène des céréales en Europe et dans les régions tempérées du monde. Cette moisissure appartient au genre Fusarium qui est l’un des genres qui produit le plus de métabolites secondaires, dont de nombreuses mycotoxines. Cependant, les connaissances sur son génome laissent présager que F. graminearum produit une très grande variété de métabolites qui sont encore inconnus. C’est pourquoi l’objectif de cette thèse a été de caractériser le métabolome de cette moisissure en identifiant de nouveaux métabolites ainsi qu’en étudiant leur toxicité. Pour cela, une méthode analytique combinant la spectrométrie de masse de haute résolution et le marquage des métabolites secondaires avec des isotopes stables a été développée. Celle‐ci à tout d’abord était validée avec le métabolome bien connu d’Aspergillus fumigatus avant d’être appliquée au métabolome de F. graminearum. Cette méthode a permis d’identifier 37 nouveaux métabolites, dont les fusaristatines A, C et D qui on été détectées pour la première fois chez F. graminearum. Le deoxynivalenol (DON) est une mycotoxine majeure produite par F. graminearum, c’est pourquoi sa toxicité a été comparée in vivo sur un modèle porcin à celle d’un mélange de fusariotoxines. Les résultats de cette étude ont montré une toxicité bien différente du mélange par rapport au DON seul, que ce soit par rapport à la performance des animaux ou à leur réponse immunitaire. Ceci suggère que certaines des fusariotoxines présentes dans le mélange auraient des répercutions biologiques et donc que leur toxicité reste encore à être élucidée. De plus, l’importance de la caractérisation de la toxicité du métabolome de F. graminearum a été soulignée par les résultats obtenus dans une seconde étude, qui ont révélés une possible implication du DON dans les maladies inflammatoires chroniques de l’intestin par l’intermédiaire des lymphocytes Th17. / F. graminearum is one of the most predominant cereal pathogen in Europe and temperate regions of the world. It belongs to the genera Fusarium, which is one of the most important in terms of the production of secondary metabolites, among which stand several mycotoxins. However, recent advances on the study of its genome suggest that only a small fraction of the secondary metabolites produced by F. graminearum have been identified so far. Therefore, the main objective of this thesis was to characterize the metabolome of this fungus by identifying new metabolites and by analyzing their toxicity. First, an analytical method combining high resolution mass spectrometry and isotopically labeled secondary metabolites was developed. The method was validated with the well known metabolome of Aspergillus fumigatus before it was applied to F. graminearum. As a result, 37 new metabolites were identified such as fusaristatin A, C and D, which were detected for the first time in the metabolome of F. graminearum. Deoxynivalenol (DON) is one of the major mycotoxins produced by this fungus. Therefore, its toxicity was compared to that of a blend of fusariotoxins in vivo in a pig model. The results of this study showed different effects of the blend compared to DON alone on animal performances and immune response. This suggests that some of the fusariotoxins present in the blend could be biologically active and their toxicity should be further investigated. Toxicity of the metabolome of F. graminearum has also been highlighted by the results of a second study which showed a possible implication of DON on the appearance of inflammatory bowel diseases by the action of Th17 lymphocytes.