RESUMO
Three strains of Pseudomonas aeruginosa were isolated: wild-type (WT, NO4) showed normal quorum sensing (QS), whereas QSD3 and QSD7 were QS-deficient (QSD) containing limited N-butyryl homoserine lactone (C4-HSL). The autoinducer activity produced by NO4 was found to be at least 50-fold higher than those by the QSD3 and the QSD7 strains. The QSDs produced lower levels of phenazine compounds (pyocyanin), siderophores (pyoverdine) and biosurfactants (rhamnolipids) than NO4. Therefore, the swarming motility and the swimming motility of the QSD3 and the QSD7 strains also decreased. Treatment with exogenous C4-HSL completely restored rhamnolipid production in both QSDs, suggesting that the biosynthesis of C4-HSL is defective. However, the biofilm production of the QSDs reached much higher levels than those of wild-types (NO4 and P. aeruginosa PAO1). And both QSD strains were more resistant than wild-type cell (NO4) against kanamycin and tobramycin. The RpoS gene, which function is related with QS, is point-nonsense mutated in QSD3 strain. But eleven QS-related genes in QSD3 were not mutated, compared to those of PAO1, which carries intact QS genes and is used as a positive control. This study is helpful in the development of novel approaches in the treatment of P. aeruginosa infections.
Assuntos
Pseudomonas aeruginosa/genética , Pseudomonas aeruginosa/metabolismo , Percepção de Quorum/genética , Percepção de Quorum/fisiologia , 4-Butirolactona/análogos & derivados , 4-Butirolactona/metabolismo , Proteínas de Bactérias/genética , Proteínas de Bactérias/fisiologia , Biofilmes/crescimento & desenvolvimento , DNA Bacteriano/genética , Farmacorresistência Bacteriana Múltipla , Regulação Bacteriana da Expressão Gênica , Genes Bacterianos/genética , Glicolipídeos/metabolismo , Canamicina/farmacologia , Oligopeptídeos/metabolismo , Infecções por Pseudomonas , Pseudomonas aeruginosa/efeitos dos fármacos , Piocianina/metabolismo , Análise de Sequência , Análise de Sequência de DNA , Fator sigma/genética , Fator sigma/fisiologia , Tobramicina/farmacologia , Fatores de Virulência/genéticaRESUMO
In a previous report, the pepper receptor-like kinase 1 (CaRLK1) gene was shown to be responsible for negatively regulating plant cell death caused by pathogens via accumulation of superoxide anions. Here, we examined whether this gene also plays a role in regulating cell death under abiotic stress. The total concentrations of free amino acids in CaRLK1-overexpressed cells (RLKox) increased by twofold compared with those of the wild-type Nicotiana tabacum BY-2 cells. Additionally, alanine and pyruvate concentrations increased by approximately threefold. These accumulations were associated with both the expression levels of the isocitrate lyase (ICL) and malate synthase genes and their specific activities, which were preferentially up-regulated in the RLKox cells. The expression levels of ethylene biosynthetic genes (ACC synthase and ACC oxidase) were suppressed, but those of both the metallothionein and lesion simulating disease 1 genes increased in the RLKox cells during submergence-induced hypoxia. The specific activity of catalase, which is involved in protecting ICL from reactive oxygen species, was also induced threefold in the RLKox cells. The primary roots of the transgenic plants that were exposed to hypoxic conditions grew at similar rates to those in normal conditions. We propose that CaRLK1 maintains a persistent hypoxia-resistant phenotype.