In Situ Gene Expression in Mixed-Culture Biofilms: Evidence of Metabolic Interactions between Community Members

• DOI: 10.1128/aem.64.2.721-732.1998
• Publication Date: 2019-12-19T19:31:36Z
• Authors: Søren Møller; Claus Sternberg; Jens Bo Andersen; Bjarke Bak Christ...; Juan Luis Ramos; Michael Givskov; Søren Molin
• Keywords: Pseudomonas putida; Expression cassette

Microbial communities growing in laboratory-based flow chambers were investigated order to study compartmentalization of specific gene expression. Among the community members studied, focus was particular on Pseudomonas putida and a strain an Acinetobacter sp., genes studied are involved biodegradation toluene related aromatic compounds. The upper-pathway promoter (Pu) meta-pathway (Pm) from TOL plasmid fused independently coding for green fluorescent protein (GFP), expression these promoters P. putida, which dominant member. Biofilms cultured chambers, combination with scanning confocal laser microscopy allowed direct monitoring activity single-cell spatial resolution. Expression Pu homogeneously induced by benzyl alcohol both pure-culture biofilms, while Pm mixed but not biofilm. By sequentially adding members, induction shown be consequence metabolic interactions between species putida. Furthermore, fixed biofilm samples organism identity determined visualized at same time combining GFP situ hybridization fluorescence-labeled 16S rRNA targeting probes. This techniques is powerful approach investigating structure-function relationships microbial communities.