Abstract
We previously demonstrated that in a metasystem consisting of Arabidopsis seedlings growing in liquid medium (in 96 well plates) even microbes considered to be innocuous such as laboratory strains of E. coli and B. subtilis can cause potent damage to the host. We further posited that such environment-induced adaptations are brought about by ‘system status changes’ (rewiring of pre-existing cellular signaling networks and components) of the host and the microbe, and that prolongation of such a situation could lead to the emergence of pathogenic states in real-life. Here, using this infection model, we show that the master regulator GacA of the human opportunistic pathogen P. aeruginosa (strain PA14) is dispensable for pathogenesis, as evidenced by three independent read-outs. The gene expression profile of the host after infection with wild type PA14 or the gacA mutant are also identical. GacA normally acts upstream of the quorum sensing regulatory circuit (that includes the regulator LasR) that controls a subset of virulence factors. Double mutants in gacA and lasR behave similar to the lasR mutant, as seen by abrogation of a characteristic cell type specific host cell damage caused by PA14 or the gacA mutant. This indicates that a previously unrecognized regulatory mechanism is operative under these conditions upstream of LasR. In addition, the detrimental effect of PA14 on Arabidopsis seedlings is resistant to high concentrations of the aminoglycoside antibiotic gentamicin. These data suggest that the Arabidopsis seedling infection system could be used to identify anti-infectives with potentially novel modes of action.
Highlights
New and re-emerging microbial diseases are a serious threat to human and agricultural health
We previously reported that PA14 causes near complete abrogation of a constitutively expressed luciferase reporter gene in Arabidopsis seedlings by five days after infection [7]
As had been shown with PA14 earlier [7], Sytox GreenH staining in PA14DgacA-infected leaves was characteristically present in most stomatal guard cells and unexpectedly the staining was evenly distributed throughout these cells (Fig. 1B)
Summary
New and re-emerging microbial diseases are a serious threat to human and agricultural health. We posited that ‘system status changes’ during the early stages of certain environmentally-induced host-microbe interactions (rewiring of pre-existing cellular signaling networks and components) may underlie the ability of a microbe to establish a new pathogenic interaction with a host, which from the host’s perspective can be viewed as an environmentally-induced ‘‘maladaptaion’’. Continued existence of such states could lead to emergence of new infectious agents [7]. Zoonotic infections, hygiene related emergence of infectious agents, and niche alteration induced diseases by indigenous microbes are likely to emerge in this manner
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