Expressão quantitativa de genes de Phytophthora parasitica e de citros durante a interação.

Abstract

The gummosis, mainly caused by the oomycete Phytophthora parasitica, is one of the most serious diseases affecting citrus crops worldwide. During the interaction, plants induce signaling cascades in order to induce defense responses. However, P. parasitica secrets effector proteins capable of modulating these host responses in order to promote the infection. In Citrus genus, commercially important species are susceptible to infection by this pathogen and the gummosis resistance is achieved in Poncirus trifoliata citrus species. Considering the lack of information on citrus-P. parasitica pathosystem, this study aimed to analyze, through RT-qPCR, the quantitative expression of P. parasitica effector and citrus defense genes during citrus-P. parasitica susceptible and resistant interactions, with Citrus sunki and P. trifoliata, respectively. As results, P. parasitica was able to recognize among susceptible or resistant host and selectively synthesize which effectors and in that intensity should be expressed. Of the 17 studied effector genes, 10 showed quantitative relative differential expression at significance level induced in P. parasitica after inoculation in trifoliate orange roots, being 06 apoplastics and 04 cytosolics. The expression profiles for the 17 effector genes in P. parasitica had two maximum peaks of expression, that are indicative of de novo synthesis of these genes along the time points of interaction, showing transcript accumulation earlier on P. trifoliata (at 6 h.a.i.) and later on C. sunki (at 96 h.a.i.). High levels of the effector gene expression in P. parasitica induced by C. sunki at 96 h.a.i. must match the necrotrophic phase of life of this oomycete, consequently due to their successful penetration into the susceptible plant cells and pathogen biomass accumulation. The presence of intracellular hyphae in cortex of C. sunki roots was abundantly visualized in the micrographs at 96 h.a.i., which may occur as a result of the plant susceptibility to the pathogen. Six hierarchical groups of co-regulated genes were formed from the expression profiles of the 17 effector genes in P. parasitica, which are grouped differently according to interact with C. sunki or P. trifoliata, indicating that the pathogen was able to recognize between susceptible or resistant host and selectively synthesize which effectors and in that intensity should be segregated. The roots of C. sunki expressed 10 components of the cascade resistance mediated by SA in response not successful to P. parasitica infection. The suppression by P. parasitica of the expression of 05 genes of the cascade resistance mediated by SA was found in P. trifoliata roots, and must indicate pathogen attempts to circumvent with the immunity of the plant. However, P. trifoliata resistance to P. parasitica should not use genes involved in the resistance cascade mediated by SA, but instead PR-5 and callose synthase genes, involving biochemical and estructural barriers. In conclusion, this study provides a new insight into the understanding of the effectors of modulation process of P. parasitica in susceptible and resistant interactions and how these hosts respond through interaction.