Caracterização funcional e envolvimento do operon htrA-XAC3983 na patogenicidade e virulência de Xanthomonas citri subsp. citri
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Data
2019-07-30
Autores
Silva, Ana Carolina Buzinari da [UNESP]
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Universidade Estadual Paulista (Unesp)
Resumo
O cancro cítrico tem como agente causal a bactéria Xanthomonas citri subsp. citri (Xac), que afeta todas as espécies de citros economicamente importantes. O sequenciamento do genoma do isolado 306 desta bactéria (Xac 306) revelou que 37,4 % dos genes não tinham uma função predita. Assim, a genômica funcional da interação Xac-hospedeiro tem um papel importante na elucidação da função dos genes e dos mecanismos envolvidos na patogenicidade e virulência dessa bactéria. Os genes XAC3981, XAC3982 e XAC3983 fazem parte deste contexto de genes codificadores de proteínas sem função conhecida (hipotéticas). As proteínas codificadas por estes três genes são conservadas em bactérias da família Xanthomonadaceae. Em um estudo prévio, o gene XAC3981 da Xac 306 foi interrompido por inserção do transposon Tn5 e esse mutante apresentou ausência de sintomas quando inoculado em plantas cítricas, sugerindo que possui um papel importante na patogenicidade de Xac. No presente estudo demonstrou-se que os genes XAC3981-3983, juntamente com o gene htrA (XAC3980), constituem um operon funcional de Xac 306. O presente estudo avaliou também os efeitos de mutações por deleção em algumas regiões do operon htrA-3983. Um mutante foi produzido por deleção de 84% da sequência codificadora do gene XAC3982 (ΔXAC3982) e outros três foram obtidos pela deleção das regiões promotoras desse operon. O mutante ΔXAC3982, quando comparado com a estirpe selvagem, alterou o fenótipo em genótipos resistentes e suscetíveis de citros e produziu menos biofilme in vitro. Observou-se que os promotores preditos, quando analisados isoladamente, não tem efeito sobre a virulência total dessa bactéria e não interferem na transcrição deste operon. Os resultados obtidos sugerem que o operon, denominado neste estudo como operon htrA-XAC3983, está relacionado com a virulência e patogenicidade de Xac.
The bacterium Xanthomonas citri subsp. citri (Xac) is the causal agent of citrus canker, a disease that affects all economically important citrus species. The genome sequencing of the 306 isolate of this bacterium (Xac 306) revealed that 37.4% of the genes had no predicted function. Thus, the functional genomics of the Xac-host interaction play an important role in the elucidation of the function of the genes and in the mechanisms involved in the pathogenicity and virulence of this bacterium. The genes XAC3981, XAC3982 and XAC3983 are part of this context of protein coding genes with unknown function (hypothetical). The proteins encoded by these three genes are conserved and present exclusively in bacteria of the family Xanthomonadaceae. In a previous study, the ORF XAC3981 from Xac306 was disrupted by insertion of the Tn5 transposon and the mutant showed no symptoms when inoculated in citrus plants, suggesting that it plays an important role in Xac pathogenicity. In the present study, genes XAC3981-3983, together with gene htrA (XAC3980), have been shown to constitute a functional operon of Xac 306. The present study also evaluated the effects of deletion mutations in some regions of XAC3980-3983 operon. One mutant was produced by deleting 84% of the coding sequence of the gene XAC3982(ΔXAC3982) and another three were obtained by deleting the promoter regions of that operon. The ΔXAC3982 mutant, when compared to the wild type strain, altered the phenotype in resistant citrus genotypes and produced less biofilm in vitro. It has been observed that the predicted promoters, when analyzed alone, have no effect on the total virulence of this bacterium and do not interfere in the transcription of this operon. The results presented here suggest that the operon, termed as operon htrA-XAC3983 in this study, is related to the virulence and pathogenicity of Xac.
The bacterium Xanthomonas citri subsp. citri (Xac) is the causal agent of citrus canker, a disease that affects all economically important citrus species. The genome sequencing of the 306 isolate of this bacterium (Xac 306) revealed that 37.4% of the genes had no predicted function. Thus, the functional genomics of the Xac-host interaction play an important role in the elucidation of the function of the genes and in the mechanisms involved in the pathogenicity and virulence of this bacterium. The genes XAC3981, XAC3982 and XAC3983 are part of this context of protein coding genes with unknown function (hypothetical). The proteins encoded by these three genes are conserved and present exclusively in bacteria of the family Xanthomonadaceae. In a previous study, the ORF XAC3981 from Xac306 was disrupted by insertion of the Tn5 transposon and the mutant showed no symptoms when inoculated in citrus plants, suggesting that it plays an important role in Xac pathogenicity. In the present study, genes XAC3981-3983, together with gene htrA (XAC3980), have been shown to constitute a functional operon of Xac 306. The present study also evaluated the effects of deletion mutations in some regions of XAC3980-3983 operon. One mutant was produced by deleting 84% of the coding sequence of the gene XAC3982(ΔXAC3982) and another three were obtained by deleting the promoter regions of that operon. The ΔXAC3982 mutant, when compared to the wild type strain, altered the phenotype in resistant citrus genotypes and produced less biofilm in vitro. It has been observed that the predicted promoters, when analyzed alone, have no effect on the total virulence of this bacterium and do not interfere in the transcription of this operon. The results presented here suggest that the operon, termed as operon htrA-XAC3983 in this study, is related to the virulence and pathogenicity of Xac.
Descrição
Palavras-chave
Cancro (Fitopatologia), Microorganismos fitopatogênicos, Biologia molecular