Caracterização fenotípica e funcional de mutantes da bactéria fitopatogênica Xanthomonas citri subsp. citri
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Data
2009-11-06
Autores
Ferreira, Cristiano Barbalho [UNESP]
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Universidade Estadual Paulista (Unesp)
Resumo
Dentro da comercialização de frutas, a citricultura é a mais importante. Representa para muitos países, dentre eles, os EUA e o Brasil, uma importante atividade econômica. Porém, esta atividade, vem sofrendo com inúmeras doenças e/ou pragas como a doença do Cancro Cítrico Asiático causada pela bactéria Xanthomonas citri subsp. citri (X. citri). Deste modo, com o objetivo do estudo do genoma funcional da X. citri, um banco de mutantes deste microorganismo foi obtido por meio de inserção aleatória do transposon Tn5, nas quais foram selecionados 53 mutantes que apresentaram, de forma superficial, alterações fenotípica em relação à X. citri selvagem. Para uma avaliação mais precisa, eles passaram por uma nova confirmação de suas alterações fenotípicas, onde foram inoculados em folhas de Citrus sinensis (Laranjeira pêra-Rio) e Citrus limonia (limoeiro cravo) e monitorados durante 16 dias, e aqueles que apresentaram as maiores alterações em relação à selvagem, tiveram confeccionadas para si curvas de crescimento in vivo. Conseguiu-se, desta forma, avaliar quantitativamente o processo patogênico, relacionar seus sintomas com dados numéricos e ainda descobrir detalhes até então não conhecidos. O mapeamento, dos respectivos loci mutados, foi realizado por seqüenciamento de DNA a partir do transposon, demonstrando que a metodologia empregada para a obtenção dos mutantes foi eficiente, conseguiu também revelar diversas proteína ainda hipotéticas, e outras, até então, não considerados como implicados no processo patogênico, como, uma Integrase, Fe-S oxidoredutase, Helicase IV, Receptor Dependente de Ton-B, entre outros
Concerning the commercialization of fruits, the citrus production is the most important. It represents for many countries, amongst them, U.S.A. and Brazil, an important economic activity. However, this activity has been suffering with many illnesses and/or plagues as the illness from the Asiatic citrus canker caused by the Xanthomonas citri subsp. citri bacterium (X. citri). In this way, from a bank of mutants of X. citri, gotten by means of random insertion of commercial one derived from transposon Tn5, had been selected 53 mutants that had presented, of superficial form, some type of phenotype alteration in relation to the wild X. citri. To a more necessary evaluation, each one of them passed for a new confirmation of its phenotype alterations, where they had been inoculated in leafs of Citrus sinensis ('Pera' sweet orange) and Citrus limonia ('Rangpur' lime) and monitored during 16 days, and those that had presented the biggest alterations in relation to the savage, had confectioned for itself in planta growth curves. We obtain, in such a way, to evaluate quantitatively the pathogenic process, to relate its symptoms with numerical data and still to discover not known details until then. The mapping of respective locus mutated was carried through by sequencing of DNA from transposon, demonstrating that the methodology used for the attainment of the mutants was efficient and still to disclose diverse genes still hypothetical, and others, until then, not considered as implied in the pathogenic process, as, Integrase, Fe-S oxidoredutase, Helicase IV, TonB-dependent receptor, among others
Concerning the commercialization of fruits, the citrus production is the most important. It represents for many countries, amongst them, U.S.A. and Brazil, an important economic activity. However, this activity has been suffering with many illnesses and/or plagues as the illness from the Asiatic citrus canker caused by the Xanthomonas citri subsp. citri bacterium (X. citri). In this way, from a bank of mutants of X. citri, gotten by means of random insertion of commercial one derived from transposon Tn5, had been selected 53 mutants that had presented, of superficial form, some type of phenotype alteration in relation to the wild X. citri. To a more necessary evaluation, each one of them passed for a new confirmation of its phenotype alterations, where they had been inoculated in leafs of Citrus sinensis ('Pera' sweet orange) and Citrus limonia ('Rangpur' lime) and monitored during 16 days, and those that had presented the biggest alterations in relation to the savage, had confectioned for itself in planta growth curves. We obtain, in such a way, to evaluate quantitatively the pathogenic process, to relate its symptoms with numerical data and still to discover not known details until then. The mapping of respective locus mutated was carried through by sequencing of DNA from transposon, demonstrating that the methodology used for the attainment of the mutants was efficient and still to disclose diverse genes still hypothetical, and others, until then, not considered as implied in the pathogenic process, as, Integrase, Fe-S oxidoredutase, Helicase IV, TonB-dependent receptor, among others
Descrição
Palavras-chave
Xanthomonas, Cítricos, Genoma funcional, PGPB, Plant pathogenic bacteria, Citrus canker, Growth curves in plant, Functional genomics, Pathogen - host interaction
Como citar
FERREIRA, Cristiano Barbalho. Caracterização fenotípica e funcional de mutantes da bactéria fitopatogênica Xanthomonas citri subsp. citri. 2009. viii, 76 f. Dissertação (mestrado) - Universidade Estadual Paulista, Faculdade de Ciências Agrárias e Veterinárias, 2009.