Abstract

Burkholderia glumae is a motile plant pathogenic bacterium that has multiple polar flagella and one LuxR/LuxI-type quorum sensing (QS) system, TofR/TofI. A QS-dependent transcriptional regulator, QsmR, activates flagellar master regulator flhDC genes. FlhDC subsequently activates flagellar gene expression in B. glumae at 37°C. Here, we confirm that the interplay between QS and temperature is critical for normal polar flagellar morphogenesis in B. glumae. In the wild-type bacterium, flagellar gene expression and flagellar number were greater at 28°C compared to 37°C. The QS-dependent flhC gene was significantly expressed at 28°C in two QS-defective (tofI::Ω and qsmR::Ω) mutants. Thus, flagella were present in both tofI::Ω and qsmR::Ω mutants at 28°C, but were absent at 37°C. Most tofI::Ω and qsmR::Ω mutant cells possessed polar or nonpolar flagella at 28°C. Nonpolarly flagellated cells processing flagella around cell surface of both tofI::Ω and qsmR::Ω mutants exhibited tumbling and spinning movements. The flhF gene encoding GTPase involved in regulating the correct placement of flagella in other bacteria was expressed in QS mutants in a FlhDC-dependent manner at 28°C. However, FlhF was mislocalized in QS mutants, and was associated with nonpolar flagellar formation in QS mutants at 28°C. These results indicate that QS-independent expression of flagellar genes at 28°C allows flagellar biogenesis, but is not sufficient for normal polar flagellar morphogenesis in B. glumae. Our findings demonstrate that QS functions together with temperature to control flagellar morphogenesis in B. glumae.

Highlights

  • Structural and physiological adaptations of bacteria facilitate their survival under various stressful environmental conditions

  • Flagellar number depends on temperature in the wildtype BGR1 To determine whether temperature is involved in B. glumae flagellar formation, the flagellar numbers of individual cells in the wild-type BGR1 were counted on Transmission electron microscopy (TEM) images

  • To determine whether the expression of flagellar genes is more elevated at 28uC compared to 37uC, we first measured the expression of the flagellar master regulator flhC gene in the wildtype BGR1, the two QS-defective (tofI)::V mutant BGS2, and the qsmR::V mutant BGS9 at 28uC

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Summary

Introduction

Structural and physiological adaptations of bacteria facilitate their survival under various stressful environmental conditions. Such adaptations often require the coordination of multiple regulatory systems to control the expression of related genes [1]. Three types of flagellar morphology (polar, peritrichous, and lateral) are determined by a combination of bacterial genetic factors and certain environmental conditions [7,8,9]. Pseudomonas aeruginosa and Vibrio cholerae possess single polar flagella, whereas Escherichia coli and Salmonella enterica have peritrichous flagella [7], and Selenomonas ruminantium has lateral flagella [8]. Vibrio parahaemolyticus has two flagellar systems, one for a single polar flagellum and one for lateral flagella, with these systems being activated by different growth conditions [9]

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