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Previous issue date: 2015-06-18 / Sclerotinia sclerotiorum, the causal agent of white mold, is a plant pathogenic fungus that has a characteristic feature, production of sclerotia, resistance structures that can be viable for up to 10 years in the soil, with ability to initiate a new cycle of infection under favorable conditions. The disease control methods are based on integrated management practices, including biological and chemical control. The proteomic study of this interaction may be a strategy for studying this pathosystem, however, the small amount of mycelium produced during the infectious process in vivo greatly limits the study of this pathosystem using this strategy. The main goals of this work were the development and validation of a culture medium that would partially mimic the host and allow the production of large amounts of micelium for in vitro studies of this pathosystem. For this purpose, a protocol was established for the production of culture media prepared with leaf extract of the hosts. These media were inoculated with sclerotia of the monosporic isolate SS 200 of S. sclerotiorum and the differential expression of effectors and candidate effectors of virulence of the fungus was evaluated by qPCR. The results showed that a large amount of micelia grew in the media and effector genes and candidate effector genes were induced in these media. These results indicate that the proposed culture media can be used to study the in vitro interaction between S. sclerotiorum and several plant hosts and that they can be useful especially in proteomic studies. / Sclerotinia sclerotiorum, agente causal do mofo branco, ?? um fungo fitopatog??nico que possui uma caracter??stica marcante, a produ????o de escler??dios, estruturas de resist??ncia que podem ser vi??veis por at?? 10 anos no solo, com capacidade de iniciar um novo ciclo de infec????o em condi????es favor??veis. Os m??todos de controle da doen??a baseiam-se em pr??ticas de manejo integrado, incluindo controle biol??gico e qu??mico. O estudo prote??mico desta intera????o pode ser uma estrat??gia para o estudo deste patossistema, entretanto, a pequena quantidade de mic??lio produzido pelo pat??geno durante o processo infeccioso in vivo limita bastante o estudo deste patossistema utilizando esta estrat??gia. Os objetivos deste trabalho foram o desenvolvimento e valida????o de um meio de cultura que mimetizasse parcialmente o hospedeiro e permitisse a produ????o de grande quantidade de massa micelial para estudos in vitro desse patossistema. Para tal, foi estabelecido um protocolo para a produ????o de meios de cultura, preparados a partir de extratos foliares dos hospedeiros. Estes meios foram inoculados com escler??dios do isolado monosp??rico de S. sclerotiorum SS 200 e a express??o diferencial de genes efetores e candidatos a efetores de virul??ncia do fungo foi avaliada por qPCR. Os resultados mostraram que em todos os meios testados houve crescimento de grande quantidade de massa micelial e que os meios foram capazes de induzir a express??o de genes efetores e candidatos a efetores. Portanto, os resultados indicam que os meios propostos podem ser usados no estudo da intera????o in vitro de S. sclerotiorum com v??rios hospedeiros, especialmente em estudos prote??micos.
| Identifer | oai:union.ndltd.org:IBICT/oai:bdtd.ucb.br:tede/2133 |
| Date | 18 June 2015 |
| Creators | Maximiano, Mariana Rocha |
| Contributors | Barros, Everaldo Gon??alves de, Dias, Simoni Campos |
| Publisher | Universidade Cat??lica de Bras??lia, Programa Strictu Sensu em Ci??ncias Gen??micas e Biotecnologia, UCB, Brasil, Escola de Sa??de e Medicina |
| Source Sets | IBICT Brazilian ETDs |
| Language | Portuguese |
| Detected Language | English |
| Type | info:eu-repo/semantics/publishedVersion, info:eu-repo/semantics/masterThesis |
| Format | application/pdf |
| Source | reponame:Biblioteca Digital de Teses e Dissertações da UCB, instname:Universidade Católica de Brasília, instacron:UCB |
| Rights | info:eu-repo/semantics/openAccess |
| Relation | -2827197273900952156, 500, 500, 600, -6392058866414562720, -3439178843068202161 |
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