Vibrio fischeri: Laboratory Cultivation, Storage, and Common Phenotypic Assays

Abstract

Vibrio fischeri is a non-pathogenic organism related to pathogenic Vibrio species that can be readily grown and stored with common laboratory equipment. In this article, protocols for routine growth, storage, and phenotypic assessment of V. fischeri as well as recipes for useful media are included. Specifically, this article describes procedures and considerations for growth of this microbe in complex and minimal media. It also describes assays for biofilm formation, motility, and bioluminescence, three commonly assessed phenotypes of Vibrio fischeri.

Quorum Sensing and Cyclic di-GMP Exert Control Over Motility of Vibrio fischeri KB2B1

“…For luminescence experiments, cells were grown overnight in SWT [0.5% tryptone, 0.3% yeast extract, and 70% artificial seawater (approximately 35 mM of magnesium sulfate, 7 mM of calcium chloride, 7 mM of potassium chloride, and 210 mM of sodium chloride); Yip et al, 2005 ] then subcultured into SWTO (SWT containing an additional 2% sodium chloride; Bose et al, 2007 ). For transformations, Tris-minimal medium (TMM) ( Christensen and Visick, 2020 ) was used. Polyvinylpyrrolidone (PVP) was used in a subset of luminescence experiments to increase viscosity and had an average molecular weight of ∼55,000 (MilliporeSigma, Burlington, MA, United States).…”

Genetic Manipulation of Vibrio fischeri

“…V. fischeri is a Gram‐negative symbiont of the Hawaiian bobtail squid, E. scolopes (McFall‐Ngai, 2014; Stabb & Visick, 2013). In the laboratory, V. fischeri has been studied with respect to phenotypes that impact symbiosis, including but not limited to biofilm formation, motility, and luminescence (for procedures to perform these assays, see Christensen & Visick, 2020). Identification of the genetic determinants that shape this symbiosis has been greatly enhanced by the expansion of the genetic toolbox for this organism.…”

“…There are many available isolates of V. fischeri with distinct phenotypes, and genetic manipulation of these strains can uncover the determinants of these phenotypes. V. fischeri strain ES114 (ATCC 700601; Boettcher & Ruby, 1990) has been commonly used as a parent strain for genetic manipulation and phenotypic characterization, and thus there are established protocols for growing and performing phenotypic characterization of this strain and its mutant derivatives (Christensen & Visick, 2020). As such, the protocols described herein primarily consider this strain.…”

“…Vibrio fischeri is a Gram‐negative, comma‐shaped, bioluminescent marine bacterium (Allen & Baumann, 1971; Lyell et al., 2017). It serves as a model system for studying a variety of traits, including light production and biofilm formation as well as the regulatory processes that control these energy‐intensive phenotypes (Christensen & Visick, 2020; Tischler, Hodge‐Hanson, & Visick, 2019). Both light production and biofilm formation are linked to the symbiotic lifestyle that V. fischeri shares with its host, the Hawaiian bobtail squid Euprymna scolopes (Mandel & Dunn, 2016; Stabb & Visick, 2013).…”

“…V. fischeri has historically been considered a genetically tractable organism, but manipulation of this organism has greatly improved within the last ∼10 years, resulting in a comprehensive genetic toolbox with optimized protocols (Christensen & Visick, 2020). Consequently, V. fischeri is now a powerful model system for dissecting genetic pathways.…”