Papers by Cindy Deloney-Marino
<p>Composition of media used in this work.</p
<p><i>V</i>. <i>fischeri</i> strains and plasmids used in this stud... more <p><i>V</i>. <i>fischeri</i> strains and plasmids used in this study.</p
<p>KV4366 was grown in LBS and spotted onto LBS(-Tris) (A) or LBS(-Tris) with 425 mM NaCl (... more <p>KV4366 was grown in LBS and spotted onto LBS(-Tris) (A) or LBS(-Tris) with 425 mM NaCl (B), 35 mM MgSO<sub>4</sub> (C), 5 mM CaCl<sub>2</sub> (D), or LBS-Opt (E). Images were captured at the same magnification at the indicated times, and asterisks indicate the time at which wrinkling became visible. At the last time point colonies were disturbed with a toothpick. Images in parts A-D are selections from Figs <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0169521#pone.0169521.g002" target="_blank">2</a>, <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0169521#pone.0169521.g004" target="_blank">4</a>, <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0169521#pone.0169521.g005" target="_blank">5</a> and <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0169521#pone.0169521.g006" target="_blank">6</a>. To measure <i>syp</i> locus transcription, Δ<i>sypA</i> mutant strains carrying a P<sub><i>sypA-lacZ</i></sub> transcriptional reporter and a plasmid overproducing RscS (pKG11/Δ<i>sypA</i>) or vector control (pKV69/Δ<i>sypA</i>) were grown in LBS(-Tris) or LBS-Opt at 24°C for 24 h and assayed for β-galactosidase activity (F). Error bars represent the standard deviation of at least three biological replicates. *, <i>P</i><0.05. To assess the effect of LBS-OPT on growth, KV4366 was grown with shaking at 24°C (blue circles) and 28°C (red triangles) in LBS(-Tris) (solid line) or LBS-Opt (dotted line), and the absorbance at 600 nm was measured each hour (G).</p
<p>KV4366 was grown in LBS and spotted onto medium containing 1.0% tryptone, 0.5% yeast ext... more <p>KV4366 was grown in LBS and spotted onto medium containing 1.0% tryptone, 0.5% yeast extract, 342 mM NaCl, and 0–10 mM CaCl<sub>2</sub>. Colonies were imaged at the same magnification at the indicated times and disturbed with a toothpick at the last time point. Asterisks indicate the first time point at which wrinkling became visible.</p
<p>KV4366 was grown in LBS and spotted onto medium containing 1.0% tryptone, 0.5% yeast ext... more <p>KV4366 was grown in LBS and spotted onto medium containing 1.0% tryptone, 0.5% yeast extract, and 210–500 mM NaCl as indicated. Colonies were imaged at the same magnification at the indicated time points and disturbed with a toothpick at the last time point. Asterisks indicate the first time point at which wrinkling became visible.</p
<p>KV4366 was grown in LBS and spotted onto medium containing only 342 mM NaCl and tryptone... more <p>KV4366 was grown in LBS and spotted onto medium containing only 342 mM NaCl and tryptone (A) or yeast extract (B) at amounts ranging from 0.25–1.0%, as indicated. Images were captured at the same magnification after 24 or 48 h, and at the last time point colonies were disturbed with a toothpick.</p
<p>KV4366 was grown in LBS and diluted to an OD<sub>600</sub> of 0.3 in LBS(-Tr... more <p>KV4366 was grown in LBS and diluted to an OD<sub>600</sub> of 0.3 in LBS(-Tris) and LBS(-Tris) with 425 mM NaCl, 35 mM MgSO4, or 5 mM CaCl2, and LBS-Opt. Triplicate cultures were grown statically in the center wells of a 24-well plate at 24°C and imaged after 20, 24, and 48 h. One sample of the triple cultures was disrupted with a toothpick at each of the three time points; for each time point, images of the undisrupted and disrupted pellicles are presented from left to right.</p
<p>Cultures of ES114 (wild-type, non-biofilm induced) and KV4366 (RscS-overproducing, biofi... more <p>Cultures of ES114 (wild-type, non-biofilm induced) and KV4366 (RscS-overproducing, biofilm-induced) were grown in LBS and spotted onto LBS or SWT. Images were captured at the same magnification following growth for 48 h.</p
for cheR of Vibrio fischeri in the Vibrio–squid symbiosis
The bacterium Vibrio fischeri requires bacterial motility to initiate colonization of the Hawaiia... more The bacterium Vibrio fischeri requires bacterial motility to initiate colonization of the Hawaiian squid Euprymna scolopes. Once colonized, however, the bacterial population becomes largely unflagellated. To understand environmental influences on V. fischeri motility, we investigated migration of this organism in tryptone-based soft agar media supplemented with different salts. We found that optimal migration required divalent cations and, in particular, Mg 2؉. At concentrations naturally present in seawater, Mg 2؉ improved migration without altering the growth rate of the cells. Transmission electron microscopy and Western blot experiments suggested that Mg 2؉ addition enhanced flagellation, at least in part through an effect on the steady-state levels of flagellin protein.
PLOS ONE, 2017
Vibrio fischeri, a marine bacterium and symbiont of the Hawaiian bobtail squid Euprymna scolopes,... more Vibrio fischeri, a marine bacterium and symbiont of the Hawaiian bobtail squid Euprymna scolopes, depends on biofilm formation for successful colonization of the squid's symbiotic light organ. Here, we investigated if culture conditions, such as nutrient and salt availability, affect biofilm formation by V. fischeri by testing the formation of wrinkled colonies on solid media. We found that V. fischeri forms colonies with more substantial wrinkling when grown on the nutrient-dense LBS medium containing NaCl relative to those formed on the more nutrient-poor, seawater-salt containing SWT medium. The presence of both tryptone and yeast extract was necessary for the production of "normal" wrinkled colonies; when grown on tryptone alone, the colonies displayed a divoting phenotype and were attached to the agar surface. We also found that the type and concentration of specific seawater salts influenced the timing of biofilm formation. Of the conditions assayed, wrinkled colony formation occurred earliest in LBS(-Tris) media containing 425 mM NaCl, 35 mM MgSO 4 , and 5 mM CaCl 2. Pellicle formation, another measure of biofilm development, was also enhanced in these growth conditions. Therefore, both nutrient and salt availability contribute to V. fischeri biofilm formation. While growth was unaffected, these optimized conditions resulted in increased syp locus expression as measured by a P sypA-lacZ transcriptional reporter. We anticipate these studies will help us understand how the natural environment of V. fischeri affects its ability to form biofilms and, ultimately, colonize E. scolopes.
Canadian Journal of Microbiology, 2012
Molecular Microbiology, 2006
Successful colonization of a eukaryotic host by a microbe involves complex microbe-microbe and mi... more Successful colonization of a eukaryotic host by a microbe involves complex microbe-microbe and microbe-host interactions. Previously, we identified in Vibrio fischeri a putative sensor kinase, RscS, required for initiating symbiotic colonization of its squid host Euprymna scolopes. Here, we analysed the role of rscS by isolating an allele, rscS1, with increased activity. Multicopy rscS1 activated transcription of genes within the recently identified symbiosis polysaccharide (syp) cluster. Wild-type cells carrying rscS1 induced aggregation phenotypes in culture, including the formation of pellicles and wrinkled colonies, in a syp-dependent manner. Colonies formed by rscS1-expressing cells produced a matrix not found in control colonies and largely lost in an rscS1-expressing sypN mutant. Finally, multicopy rscS1 provided a colonization advantage over control cells and substantially enhanced the ability of wildtype cells to aggregate on the surface of the symbiotic organ of E. scolopes; this latter phenotype similarly depended upon an intact syp locus. These results suggest that transcription induced by RscSmediated signal transduction plays a key role in colonization at the aggregation stage by modifying the cell surface and increasing the ability of the cells to adhere to one another and/or to squid-secreted mucus.
Journal of Bacteriology, 2005
The bacterium Vibrio fischeri requires bacterial motility to initiate colonization of the Hawaiia... more The bacterium Vibrio fischeri requires bacterial motility to initiate colonization of the Hawaiian squid Euprymna scolopes . Once colonized, however, the bacterial population becomes largely unflagellated. To understand environmental influences on V. fischeri motility, we investigated migration of this organism in tryptone-based soft agar media supplemented with different salts. We found that optimal migration required divalent cations and, in particular, Mg 2+ . At concentrations naturally present in seawater, Mg 2+ improved migration without altering the growth rate of the cells. Transmission electron microscopy and Western blot experiments suggested that Mg 2+ addition enhanced flagellation, at least in part through an effect on the steady-state levels of flagellin protein.
Canadian Journal of Microbiology, 2012
Upon hatching, the Hawaiian squid Euprymna scolopes is rapidly colonized by its symbiotic partner... more Upon hatching, the Hawaiian squid Euprymna scolopes is rapidly colonized by its symbiotic partner, the bioluminescent marine bacterium Vibrio fischeri . Vibrio fischeri cells present in the seawater enter the light organ of juvenile squid in a process that requires bacterial motility. In this study, we investigated the role chemotaxis may play in establishing this symbiotic colonization. Previously, we reported that V. fischeri migrates toward numerous attractants, including N-acetylneuraminic acid (NANA), a component of squid mucus. However, whether or not migration toward an attractant such as squid-derived NANA helps the bacterium to localize toward the light organ is unknown. When tested for the ability to colonize juvenile squid, a V. fischeri chemotaxis mutant defective for the methyltransferase CheR was outcompeted by the wild-type strain in co-inoculation experiments, even when the mutant was present in fourfold excess. Our results suggest that the ability to perform chemota...
Applied and Environmental Microbiology, 2003
Newlyhatched juveniles of the Hawaiian squid Euprymna scolopes rapidly become colonized by the bi... more Newlyhatched juveniles of the Hawaiian squid Euprymna scolopes rapidly become colonized by the bioluminescent marine bacterium Vibrio fischeri . Motility is required to establish the symbiotic colonization, but the role of chemotaxis is unknown. In this study we analyzed chemotaxis of V. fischeri to a number of potential attractants. The bacterium migrated toward serine and most sugars tested. V. fischeri also exhibited the unusual ability to migrate to nucleosides and nucleotides as well as to N -acetylneuraminic acid, a component of squid mucus.
<p>KV4366 was grown in LBS and spotted onto medium containing 1.0% tryptone, 0.5% yeast ext... more <p>KV4366 was grown in LBS and spotted onto medium containing 1.0% tryptone, 0.5% yeast extract, 342 mM NaCl, and 0–100 mM MgSO<sub>4</sub>. Colonies were imaged at the same magnification at the indicated times and disturbed with a toothpick at the last time point. Asterisks indicate the first time point at which wrinkling became visible.</p
Typescript. Thesis (M.S.)--Eastern Washington University, 1996. Vita. Includes bibliographical re... more Typescript. Thesis (M.S.)--Eastern Washington University, 1996. Vita. Includes bibliographical references (leaves 36-38).
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Papers by Cindy Deloney-Marino