RESUMO
The bacterial pathogen Pseudomonas syringae pv tomato DC3000 suppresses plant innate immunity with effector proteins injected by a type III secretion system (T3SS). The cysteine protease effector HopN1, which reduces the ability of DC3000 to elicit programmed cell death in non-host tobacco, was found to also suppress the production of defence-associated reactive oxygen species (ROS) and callose when delivered by Pseudomonas fluorescens heterologously expressing a P. syringae T3SS. Purified His(6) -tagged HopN1 was used to identify tomato PsbQ, a member of the oxygen evolving complex of photosystem II (PSII), as an interacting protein. HopN1 localized to chloroplasts and both degraded PsbQ and inhibited PSII activity in chloroplast preparations, whereas a HopN1(D299A) non-catalytic mutant lost these abilities. Gene silencing of NtPsbQ in tobacco compromised ROS production and programmed cell death by DC3000. Our data reveal PsbQ as a contributor to plant immunity responses and a target for pathogen suppression.
Assuntos
Cisteína Proteases/metabolismo , Interações Hospedeiro-Patógeno , Fotossíntese , Complexo de Proteína do Fotossistema II/metabolismo , Pseudomonas syringae/enzimologia , Solanum lycopersicum/microbiologia , Fatores de Virulência/metabolismo , Apoptose , Proteínas de Bactérias/metabolismo , Evasão da Resposta Imune , Imunidade Inata , Solanum lycopersicum/imunologia , Doenças das Plantas/imunologia , Doenças das Plantas/microbiologia , Mapeamento de Interação de Proteínas , Pseudomonas fluorescens/genética , Pseudomonas fluorescens/patogenicidade , Pseudomonas syringae/patogenicidade , Espécies Reativas de Oxigênio/antagonistas & inibidores , Espécies Reativas de Oxigênio/metabolismoRESUMO
Antimicrobial peptides constitute an important factor in the defense of plants against pathogens, and bacterial resistance to these peptides have previously been shown to be an important virulence factor in Dickeya dadantii, the causal agent of soft-rot disease of vegetables. In order to understand the bacterial response to antimicrobial peptides, a transcriptional microarray analysis was performed upon treatment with sub-lethal concentration of thionins, a widespread plant peptide. In all, 36 genes were found to be overexpressed, and were classified according to their deduced function as i) transcriptional regulators, ii) transport, and iii) modification of the bacterial membrane. One gene encoding a uricase was found to be repressed. The majority of these genes are known to be under the control of the PhoP/PhoQ system. Five genes representing the different functions induced were selected for further analysis. The results obtained indicate that the presence of antimicrobial peptides induces a complex response which includes peptide-specific elements and general stress-response elements contributing differentially to the virulence in different hosts.
Assuntos
Peptídeos Catiônicos Antimicrobianos/farmacologia , Enterobacteriaceae/genética , Enterobacteriaceae/metabolismo , Regulação Bacteriana da Expressão Gênica/efeitos dos fármacos , Peptídeos Catiônicos Antimicrobianos/metabolismo , Enterobacteriaceae/patogenicidade , Perfilação da Expressão Gênica , Genoma Bacteriano , Mutação , Peptídeos Cíclicos , Plantas/metabolismo , Regiões Promotoras Genéticas , Ligação Proteica , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Transcriptoma , VirulênciaRESUMO
Plasmids are key vehicles of horizontal gene transfer and contribute greatly to bacterial genome plasticity. In this work, we studied a group of plasmids from enterobacteria that encode phylogenetically related mobilization functions that populate the previously non-described MOBQ 4 relaxase family. These plasmids encode two transfer genes: mobA coding for the MOBQ 4 relaxase; and mobC, which is non-essential but enhances the plasmid mobilization frequency. The origin of transfer is located between these two divergently transcribed mob genes. We found that MPFI conjugative plasmids were the most efficient helpers for MOBQ 4 conjugative dissemination among clinically relevant enterobacteria. While highly similar in their mobilization module, two sub-groups with unrelated replicons (Rep_3 and ColE2) can be distinguished in this plasmid family. These subgroups can stably coexist (are compatible) and transfer independently, despite origin-of-transfer cross-recognition by their relaxases. Specific discrimination among their highly similar oriT sequences is guaranteed by the preferential cis activity of the MOBQ 4 relaxases. Such a strategy would be biologically relevant in a scenario of co-residence of non-divergent elements to favor self-dissemination.