Continuous culture of Variovorax paradoxus biofilms on borosilicate glass
Poster Number
14C
Format
Poster Presentation (Research Day, April 30)
Faculty Mentor Name
Paul Orwin
Faculty Mentor Department
Biological Sciences
Abstract/Artist Statement
Biofilm formation is the irreversible attachment and growth of a population of microorganisms on a surface. This process includes phenotypic adaptations that alter the organism’s cell morphology, physiology, and metabolism. A major goal of our research group is to understand the underlying genetic program that leads to this phenotypic shift.
Previous studies from the Orwin lab showed that Variovorax paradoxus EPS, a Gram-negative beta proteobacterium, expressed filamentous structures within the biofilm that were not observed when grown in a static liquid medium. Treatment of these biofilms with DNAse followed by staining with a mixture of Propidium Iodide and Syto 9 showed that the filaments were eliminated by this treatment, suggesting that extracellular DNA plays an important role in the formation of biofilms. Here we report results of continuous cultures of several strains of Variovorax which formed biofilms on glass chips in a chemostat. The morphology and density of the biofilms was observed using DAPI staining on an Echo Revolve epifluorescence microscope. RNA extraction was also attempted on these cultures, and different nutrient levels were evaluated to determine the optimal growth approach yield high density, phenotypically consistent biofilms.
The overall goal of this is to compare the RNA extracted from the biofilms to similar extractions from exponential growth to identify genes specifically upregulated in these continuous flow systems. Using this strategy we can identify global regulatory systems for controlling this complex developmental process.
Location
Information Commons, William Knox Holt Memorial Library and Learning Center
Start Date
30-4-2022 10:00 AM
End Date
30-4-2022 12:00 PM
Continuous culture of Variovorax paradoxus biofilms on borosilicate glass
Information Commons, William Knox Holt Memorial Library and Learning Center
Biofilm formation is the irreversible attachment and growth of a population of microorganisms on a surface. This process includes phenotypic adaptations that alter the organism’s cell morphology, physiology, and metabolism. A major goal of our research group is to understand the underlying genetic program that leads to this phenotypic shift.
Previous studies from the Orwin lab showed that Variovorax paradoxus EPS, a Gram-negative beta proteobacterium, expressed filamentous structures within the biofilm that were not observed when grown in a static liquid medium. Treatment of these biofilms with DNAse followed by staining with a mixture of Propidium Iodide and Syto 9 showed that the filaments were eliminated by this treatment, suggesting that extracellular DNA plays an important role in the formation of biofilms. Here we report results of continuous cultures of several strains of Variovorax which formed biofilms on glass chips in a chemostat. The morphology and density of the biofilms was observed using DAPI staining on an Echo Revolve epifluorescence microscope. RNA extraction was also attempted on these cultures, and different nutrient levels were evaluated to determine the optimal growth approach yield high density, phenotypically consistent biofilms.
The overall goal of this is to compare the RNA extracted from the biofilms to similar extractions from exponential growth to identify genes specifically upregulated in these continuous flow systems. Using this strategy we can identify global regulatory systems for controlling this complex developmental process.