Submitted by LARISSA NOGUEIRA DE SOUZA null (larissaiftm@gmail.com) on 2017-05-24T19:31:43Z No. of bitstreams: 1 Dissertacao_Larissa_Nogueira_de_Souza.pdf: 3902345 bytes, checksum: fc3833c63f2f779042077b0ab45fb745 (MD5) / Approved for entry into archive by Luiz Galeffi (luizgaleffi@gmail.com) on 2017-05-24T20:56:09Z (GMT) No. of bitstreams: 1 souza_ln_me_jabo.pdf: 3902345 bytes, checksum: fc3833c63f2f779042077b0ab45fb745 (MD5) / Made available in DSpace on 2017-05-24T20:56:09Z (GMT). No. of bitstreams: 1 souza_ln_me_jabo.pdf: 3902345 bytes, checksum: fc3833c63f2f779042077b0ab45fb745 (MD5) Previous issue date: 2017-02-24 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / O clima tropical brasileiro propicia o desenvolvimento de inúmeras doenças fúngicas nas culturas produzidas, incluindo as fruteiras tropicais. O Brasil é um grande produtor de frutas, incluindo goiaba e manga, que são inclusive exportadas para outros países. Além das doenças conhecidas que incidem sobre essas culturas, sintomas atípicos têm sido observados em condições de campo, sem relação específica com patógenos já identificados. Assim, este trabalho teve por objetivo isolar fungos de tecidos lesionados de frutos, folhas e ramos de goiabeiras, assim como de folhas de mangueiras para estabelecer suas relações parasitárias e identificá-los molecularmente. Foram obtidos 25 isolados de goiabeiras de três municípios e dois isolados de mangueiras de um município do Estado de São Paulo. Os isolados tiveram sua região ITS rDNA (Internal Transcribed Spacer) e a região parcial do gene TEF1 (translation elongation factor 1-alpha) sequenciadas, o que proporcionou a identificação acurada dos isolados, em gênero. Os isolados de goiabeiras foram identificados como Neopestalotiopsis spp., sendo esse o primeiro relato em goiabeiras, e os isolados de mangueiras foram agrupados juntamente com as espécies Pseudopestalotiopsis theae e Pestalotiopsis trachicarpicola, sendo este também o primeiro relato dessas espécies em mangueiras. Os isolados de goiabeiras e de mangueiras foram inoculados em folhas destacadas de goiabeira ‘Paluma’, em condições de laboratório, com ferimento e sem ferimento. Em adição, os isolados oriundos de mangueiras foram inoculados em folhas destacadas de mangueiras ‘Tommy Atkins’. Os isolados causaram sintomas característicos daqueles dos quais provieram, sendo com maior severidade quando em folhas previamente feridas. No entanto, os isolados de Neopestalotiopsis spp. também foram capazes de ocasionar sintomas em folhas intactas. Concluiu-se que novos gêneros de fungos estão associados a lesões em mangueiras e goiabeiras no Brasil, os quais apresentam falta de especificidade de tecidos e hospedeiros, representando novo risco à sanidade das plantas. / The Brazilian tropical climate promotes the development of numerous fungal diseases in the crops produced in the country, including tropical fruit crops. Brazil is a major producer of fruits, including guava and mango, which are even exported to other countries. Besides the known diseases that affect these crops, atypical symptoms have been observed under field conditions, with no specific relation to pathogens already identified. Thus, this work aimed to isolate fungi from such lesions of fruits, leaves and branches of guava plants, as well as from leaves of mango plants to identify them molecularly and prove their symptomatic association with such crops. We obtained 25 isolates from guava plants located in three municipalities and two isolates from mango plants located in one municipality of São Paulo state. These isolates were submitted to sequencing of ITS rDNA region (Internal Transcribed Spacer) and the partial TEF1 gene (translation elongation factor 1-alpha), which provided the accurate identification of the isolates in genus. The isolates from guava plants were identified as Neopestalotiopsis spp. It is the first report of this genus in guava. Moreover, the isolates from mango plants were grouped with the species Pseudopestalotiopsis theae and Pestalotiopsis trachicarpicola. It is also the first report of these species in mango trees. Guava and mango isolates were inoculated on 'Paluma' guava leaves under laboratory conditions, with injury and without injury. In addition, the isolates from mango plants were inoculated on detached leaves of 'Tommy Atkins' mango. The isolates were able to cause symptoms similar to those from which they came in greater severity in wounded leaves. However, the isolates of Neopestalotiopsis spp. were also capable of causing symptoms on intact leaves. Thus, it was concluded that new genotypes of fungi are associated with lesions in mango and guava plants in Brazil, which present lack of tissue and host specificity, representing a new risk to the orchards sanity.

La verticilliose est causée par des agents pathogènes telluriques du genre Verticillium. Elle est, depuis sa découverte dans les années 50, maladie majeure du tournesol en Argentine où des sources de résistances ont été identifiées. En France, c’est une maladie de plus en plus fréquente, observée chaque année sur de nouvelles zones de production. Elle suscite désormais des efforts dans la recherche de moyens de lutte sur ce territoire. Ce travail s’est concentré sur l’étude des interactions Tournesol - agents causals de la verticilliose à deux niveaux d’observation : celui du système de culture (français vs. argentin) et celui de l’individu. L’objectif est d’apporter des connaissances sur l’agent causal et sur le déterminisme moléculaire dans la résistance à la verticilliose du tournesol afin de développer un modèle de criblage de résistances à grande échelle. L’étude de la maladie dans les systèmes de culture a permis de mettre en évidence l’existence d’une différence significative de la réponse du tournesol à Verticillium entre la France et l’Argentine. L’étude moléculaire des pathogènes vasculaires in planta, échantillonnés dans les 2 systèmes de culture, a permis de confirmer l’implication majeure de V. dahliae dans la verticilliose du tournesol. En conditions contrôlées, une étude comparative de la pathogénicité de plusieurs isolats de V. dahliae (de la tomate, du coton, du sol) sur le tournesol a mis en évidence que seul l’isolat 85S, isolé à partir du tournesol, est capable de le coloniser et de provoquer des symptômes. L’étude du génome de l’isolat 85S révèle que cet isolat n’appartient à aucune branche existante de l’arbre phylogénétique ; il forme un groupe per se, associé aux isolats non défoliant du coton mais infectant la tomate. L’hypothèse de la spécificité de la réponse induite dans l’hypocotyle et les feuilles du tournesol par certains isolats de V. dahliae a été confirmée en étudiant la cinétique de l’expression de 9 gènes associés à la défense, 5 semaines après inoculation. Le tournesol met uniquement en place son système de défense en réponse à l’infection par 85S. La réponse semble induite ; la colonisation n’est pas systémique, la biomasse fongique n’a pas été détectée dans l’hypocotyle et les feuilles de l’hybride asymptomatique. L’ensemble de ces travaux a conduit au développement d’un modèle pour le criblage de résistances à Verticillium chez le tournesol. Celui-ci répond aux contraintes liées à la diversité du pathogène dans les méga-environnements, conséquences de pressions de sélection différentielles. / Verticillium wilt is caused by soil-borne fungi of the genus Verticillium. From its discovery in the 50’s, sunflower Verticillium wilt is a major disease in Argentina where sources of resistances have been identified. Since few years, the disease occurs more frequently in France raising concerns on sources of resistances discoveries regarding its spread to other sunflower French production areas. This work focus on the study of Sunflower- causals agent of Verticillium wilt interaction at 2 levels of observation: at the cultural system (French vs Argentinian) and at the plant individual scale. The aim is to provide identification of the causal agents and knowledge on the molecular determinism of sunflower resistances to implement high-throughput plant phenotyping approach. Disease symptom studies within the cultural systems reveal a significant difference in the phenotypic expression of sunflower against Verticillium pressure according to the location in France or in Argentina. Molecular studies of isolates in planta, from naturally infested field in cultural systems reveal the major implication of V. dahliae in the sunflower Verticillium wilt disease. In controlled conditions, comparative studies of V. dahliae isolates pathogenicity (isolated from cotton, tomato and soil) on sunflower reveal that only 85S, isolated from sunflower is able to colonize and provoke symptoms. Genomic studies of 85S isolates reveal that the isolate did not belong to any branch of the current phylogenetic tree; 85S makes a 'per se' group within the cotton non-defoliating but tomato infecting strains. Specificity of induced responses in sunflower hypocotyl and leaves by only some of V. dahliae isolates have been confirmed through a gene expression kinetic analysis of 9 defenses related genes on 5 weeks post inoculation. Sunflower genotype is responding only to the 85S isolate. Resistance seems to be induced; colonization is not systemic as pathogen biomass has been detected but not quantify in symptomless cultivar. Our study finally leads to the implementation of Verticillium resistances screening model on the sunflower with respect to the constraints related to the pathogen diversity from the different environments, according to differential selection pressure.