Incompatibility of Vibrio fischeri Strains during Symbiosis Establishment Depends on Two Functionally Redundant <i>hcp</i> Genes

Abstract

Bacteria that have the capacity to fill the same niche will compete with one another for the space and resources available within an ecosystem. Such competition is heightened among different strains of the same bacterial species. Nevertheless, different strains often inhabit the same host. The molecular mechanisms that impact competition between different strains within the same host are poorly understood. To address this knowledge gap, the type VI secretion system (T6SS), which is a mechanism for bacteria to kill neighboring cells, was examined in the marine bacterium <i>Vibrio fischeri</i> Different strains of <i>V. fischeri</i> naturally colonize the light organ of the bobtail squid <i>Euprymna scolopes</i> The genome of FQ-A001, a T6SS-positive strain, features two <i>hcp</i> genes that are predicted to encode identical subunits of the T6SS. Coincubation assays showed that either <i>hcp</i> gene is sufficient for FQ-A001 to kill another strain via the T6SS <i>in vitro</i> Additionally, induction of <i>hcp</i> expression is sufficient to induce killing activity in an FQ-A001 mutant lacking both <i>hcp</i> genes. Squid colonization assays involving inocula of FQ-A001-derived strains mixed with ES114 revealed that both <i>hcp</i> genes must be deleted for FQ-A001 and ES114 to occupy the same space within the light organ. These experimental results provide insight into the genetic factors necessary for the T6SS of <i>V. fischeri</i> to function <i>in vivo</i>, thereby increasing understanding of the molecular mechanisms that impact strain diversity within a host.<b>IMPORTANCE</b> Different bacterial strains compete to occupy the same niche. The outcome of such competition can be affected by the type VI secretion system (T6SS), an intercellular killing mechanism of bacteria. Here an animal-bacterial symbiosis is used as a platform for study of the genetic factors that promote the T6SS-mediated killing of one strain by another. Identification of the molecular determinants of T6SS function <i>in vivo</i> contributes to the understanding of how different strains interact within a host.