Abstract

Symbiotic bacteria use diverse strategies to compete for host colonization sites. However, little is known about the environmental cues that modulate interbacterial competition as they transition between free-living and host-associated lifestyles. We used the mutualistic relationship between Eupyrmna scolopes squid and Vibrio fischeri bacteria to investigate how intraspecific competition is regulated as symbionts move from the seawater to a host-like environment. We recently reported that V. fischeri uses a type VI secretion system (T6SS) for intraspecific competition during host colonization. Here, we investigated how environmental viscosity impacts T6SS-mediated competition by using a liquid hydrogel medium that mimics the viscous host environment. Our data demonstrate that although the T6SS is functionally inactive when cells are grown under low-viscosity liquid conditions similar to those found in seawater, exposure to a host-like high-viscosity hydrogel enhances T6SS expression and sheath formation, activates T6SS-mediated killing in as little as 30 min, and promotes the coaggregation of competing genotypes. Finally, the use of mass spectrometry-based proteomics revealed insights into how cells may prepare for T6SS competition during this habitat transition. These findings, which establish the use of a new hydrogel culture condition for studying T6SS interactions, indicate that V. fischeri rapidly responds to the physical environment to activate the competitive mechanisms used during host colonization.IMPORTANCE Bacteria often engage in interference competition to gain access to an ecological niche, such as a host. However, little is known about how the physical environment experienced by free-living or host-associated bacteria influences such competition. We used the bioluminescent squid symbiont Vibrio fischeri to study how environmental viscosity impacts bacterial competition. Our results suggest that upon transition from a planktonic environment to a host-like environment, V. fischeri cells activate their type VI secretion system, a contact-dependent interbacterial nanoweapon, to eliminate natural competitors. This work shows that competitor cells form aggregates under host-like conditions, thereby facilitating the contact required for killing, and reveals how V. fischeri regulates a key competitive mechanism in response to the physical environment.

Highlights

  • Symbiotic bacteria use diverse strategies to compete for host colonization sites

  • We previously showed that the V. fischeri genome encodes a strainspecific T6SS on chromosome II (T6SS2) that is used for interstrain competition during the initial colonization of the squid host [2]

  • We sought to manipulate the viscosity of the liquid medium for use in our in vitro competition assay to better reflect the physical environments encountered by V. fischeri during habitat transition

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Summary

Introduction

Symbiotic bacteria use diverse strategies to compete for host colonization sites. little is known about the environmental cues that modulate interbacterial competition as they transition between free-living and host-associated lifestyles. The use of mass spectrometry-based proteomics revealed insights into how cells may prepare for T6SS competition during this habitat transition These findings, which establish the use of a new hydrogel culture condition for studying T6SS interactions, indicate that V. fischeri rapidly responds to the physical environment to activate the competitive mechanisms used during host colonization. The genomes of many host-associated bacteria encode diverse strategies for interbacterial competition, including the type VI secretion system (T6SS) [1,2,3,4,5,6], which is a contact-dependent killing mechanism This protein export machinery has been identified in commensals [2, 7,8,9], pathogens [1, 3,4,5,6, 10, 11], and beneficial symbionts [2, 7] and is predicted to critically impact bacterial fitness in the host [12,13,14,15,16]. Despite the prevalence of T6SSs in host-associated bacteria, few studies have investigated how the host’s physical environment impacts T6SS function [25]

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