Abstract

Quorum sensing (QS) in vitro controls production of plant cell wall degrading enzymes (PCWDEs) and other virulence factors in the soft rotting enterobacterial plant pathogen Pectobacterium atrosepticum (Pba). Here, we demonstrate the genome-wide regulatory role of QS in vivo during the Pba–potato interaction, using a Pba-specific microarray. We show that 26% of the Pba genome exhibited differential transcription in a QS (expI-) mutant, compared to the wild-type, suggesting that QS may make a greater contribution to pathogenesis than previously thought. We identify novel components of the QS regulon, including the Type I and II secretion systems, which are involved in the secretion of PCWDEs; a novel Type VI secretion system (T6SS) and its predicted substrates Hcp and VgrG; more than 70 known or putative regulators, some of which have been demonstrated to control pathogenesis and, remarkably, the Type III secretion system and associated effector proteins, and coronafacoyl-amide conjugates, both of which play roles in the manipulation of plant defences. We show that the T6SS and a novel potential regulator, VirS, are required for full virulence in Pba, and propose a model placing QS at the apex of a regulatory hierarchy controlling the later stages of disease progression in Pba. Our findings indicate that QS is a master regulator of phytopathogenesis, controlling multiple other regulators that, in turn, co-ordinately regulate genes associated with manipulation of host defences in concert with the destructive arsenal of PCWDEs that manifest the soft rot disease phenotype.

Highlights

  • Quorum sensing (QS) is a population density-dependent regulatory mechanism, utilising freely diffusible chemical signal molecules, which controls a wide range of phenotypes in many different bacteria [1]

  • We developed a whole genome microarray for Pba strain SCRI1043 (Pba1043) and report its use to study gene expression from an expI mutant of Pba1043 grown in planta, to determine global effects of QS on gene regulation during potato infection, with particular emphasis on the relationship between plant cell wall degrading enzymes (PCWDEs) and possible stealth mechanisms

  • As QS is acyl-homoserine lactone (AHL) concentration-dependent, its impact is likely to be greatest towards the latter stages of infection, where large quantities of PCWDEs are induced to attack plant cells and the characteristic soft rot disease symptoms occur [4]

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Summary

Introduction

Quorum sensing (QS) is a population density-dependent regulatory mechanism, utilising freely diffusible chemical signal molecules, which controls a wide range of phenotypes in many different bacteria [1]. These pathogens cause disease primarily through the coordinate and prolific production of a variety of plant cell wall degrading enzymes (PCWDEs), which are secreted to the extracellular environment through the Type I (protease) and Type II (pectinases and cellulases) secretion systems [4]. They possess a Type III secretion system (T3SS) with cognate effector (DspA/E) and helper/harpin proteins (HrpN/HrpW), which is required for full virulence [5]. Different strains of pectobacteria possess up to three homologues of LuxR [16] including: VirR, which plays a central role in the repression of QS-regulated

Author Summary
A Mutation in expI Reduces Virulence and OHHL Production
Conclusions
Findings
Materials and Methods
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