Metabolomics,Unknown,Transcriptomics,Genomics,Proteomics

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Effect of 3,OC12-HSL on the transcriptome of the ectomycorrhizal fungus Laccaria bicolor S238N

ABSTRACT: Bacteria use small diffusible molecules to communicate and to coordinate population response by a cell density-depend mechanism named quorum sensing (QS). If such a cross-talk was first described between bacterial species, it was recently evidenced that bacterial QS signal molecules can also be sensed by eukaryotic organisms thus playing as inter-kingdom signalling molecules. Although ectomycorrhizal fungi interact physically and metabolically with myriads of bacterial species in soils, nothing is known about the ability of ectomycorrhiza-associated bacteria to produce QS molecules and the potential impact of these molecules in the interaction. While investigating the potential effect of these molecules on the physiology of the ectomycorrhizal fungus, we found that 3O,C12-HSL also induced an alteration of the transcriptome of L. bicolor S238N. This study used the microarray design as follows: cDNA libraries from pure culture of Laccaria bicolor S238N mycelium and from three stages of L. bicolor S238N sporocarp development (Lb2 library: stipes and caps of 5–10 mm growing sporocarps; Lb3 library: caps of 30– 40 mm mature sporocarps), collected under Douglas ﬁr seedlings grown in a glasshouse, were constructed in the λTriplEx2 vector. The cDNA inserts from bacterial clones were PCR-ampliﬁed and 4992 cDNAs were arrayed from 384-well microtitre plates on Nylon membranes. Three independent biological replicates were performed for the treatment (3,OC12-HSL) and for the control (ethyl acetate)

ORGANISM(S): Laccaria bicolor S238N-H82

SUBMITTER: Francis Martin

PROVIDER: E-GEOD-18635 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Project description:Quorum-sensing (QS) is a cell-cell communication system that controls gene expression in many bacterial species, mediated by diffusible signal molecules. While the intracellular regulatory mechanisms of QS are often well-understood, the functional roles of QS remain controversial. In particular, the use of multiple signals by many bacterial species poses a serious challenge to current functional theories. Here we address this challenge by showing that bacteria can use multiple QS signals to infer both their social (density) and physical (mass-transfer) environment. Analytical and evolutionary simulation models show that the detection of and response to complex social/physical contrasts requires multiple signals with distinct half-lives and combinatorial (non-additive) responses to signal concentrations. We test these predictions using the opportunistic pathogen Pseudomonas aeruginosa, and demonstrate significant differences in signal decay between its two primary signal molecules as well as diverse combinatorial responses to dual signal inputs. QS is associated with the control of secreted factors, and we show that secretome genes are preferentially controlled by synergistic M-bM-^@M-^XAND-gateM-bM-^@M-^Y responses to multiple signal inputs, ensuring the effective expression of secreted factors in high density and low mass-transfer environments. Our results support a novel functional hypothesis for the use of multiple signals and, more generally, show that bacteria are capable of combinatorial communication. The two primary signal molecules of P. aeruginosa are the homoserine lactones N-(3-oxododecanoyl)-L-homoserine lactone (3-oxo-C12-HSL) and N-butyryl-homoserine lactone (C4-HSL). Effects of the different signal molecules was assessed using a double QS synthase mutant of Pseudomonas aeruginosa PAO1 lasI/rhlI grown at 37M-BM-0C in 25 ml LB broth and 250 ml flasks with shaking at 200 r.p.m. in four treatments, each with a replicate: (a) no addition; (b) 3-oxo- C12-HSL; (c) C4-HSL; and (d) both 3-oxo-C12-HSL and C4-HSL.

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Effect of 3,OC12-HSL on the transcriptome of the ectomycorrhizal fungus Laccaria bicolor S238N

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