Project description:In the frame of the European MELiSSA project, which aims to develop a closed biological life support system for forthcoming long term space exploration missions, the study of the alpha-proteobacterium Rhodospirillum rubrum S1H cultivated in space related environmental conditions has started. In the present work, the bacterium was grown using two different microgravity simulators, namely the Rotating Wall Vessel (RWV) and the Random Positioning Machine (RPM) and its response was evaluated at both the transcriptomic and proteomic levels using respectively a dedicated whole-genome microarray and high-througput proteomics. At the transcriptomic level, 13 genes were found significantly induced in the RWV samples and all 13 were included in the more pronounced response of 235 induced genes of R. rubrum S1H to the RPM cultivation. On the other hand, at the proteomic level, a few common proteins were found to be differentially expressed in RWV and RPM while the RWV appeared to induce a higher number of significantly regulated proteins. However, the transcriptomic and the proteomic approaches appeared to be complementary pointing out the likely interrelation between quorum sensing, cell pigmentation and cell aggregation in R. rubrum S1H. Future studies will aim to characterize this unknown quorum sensing regulon.
Project description:In the present study, the susceptibility of the purple pigmented photosynthetic alphaproteobacterium Rhodospirillum rubrum S1H to gamma irradiation was investigated and its molecular response was characterised by means of gene expression analysis. R. rubrum S1H appears to be about 4 times more sensitive than the model strain Escherichia coli MG1655 to cobalt-60 gamma irradiation. Whole genome response of R. rubrum to 25 Gy revealed the common expression of SOS response related genes in both rich and minimal media. Quantitative expression of the lexA gene was followed after various recovery time following gamma irradiation and showed differential gene expression pattern between minimal and rich medium. This work paves the way for forthcoming molecular studies on the effect of ionizing radiation on R. rubrum S1H and the other MELiSSA strains. Keywords: Rhodospirillum rubrum; ionizing radiation tolerance; microarray; quantitative PCR.
Project description:In the present study, the susceptibility of the purple pigmented photosynthetic alphaproteobacterium Rhodospirillum rubrum S1H to gamma irradiation was investigated and its molecular response was characterised by means of gene expression analysis. R. rubrum S1H appears to be about 4 times more sensitive than the model strain Escherichia coli MG1655 to cobalt-60 gamma irradiation. Whole genome response of R. rubrum to 25 Gy revealed the common expression of SOS response related genes in both rich and minimal media. Quantitative expression of the lexA gene was followed after various recovery time following gamma irradiation and showed differential gene expression pattern between minimal and rich medium. This work paves the way for forthcoming molecular studies on the effect of ionizing radiation on R. rubrum S1H and the other MELiSSA strains. Keywords: Rhodospirillum rubrum; ionizing radiation tolerance; microarray; quantitative PCR. Two-condition experiments. Comparing samples after exposure to gamma (Co-60) irradiation with a non-irradiated sample. At least biological duplicates. Each array contains 3 technical replicates.
Project description:In the context of the Micro-Ecological Life Support System Alternative (MELiSSA) project, the quorum sensing system (QS) of R. rubrum S1H has been shown to rely on acyl homoserine lactones (AHLs) as communication signals. In addition, previous studies in our lab have suggested that pigment content and photosynthetic membrane production could be under the control of QS. In the present study, the transcriptomic and proteomic profiles of a QS-deficient mutant (R. rubrum strain M68) that does not produce AHLs as the WT strain (R. rubrum S1H) were compared when cultivated in light anaerobic conditions using acetate as carbon source. Transcriptomic and proteomic approaches revealed that 330 genes and 217 proteins were differentially expressed in M68 compared to S1H, indicating that several operons were QS-regulated i.e. flagellar assembly, chemotaxis, photosynthesis, electron transport, stress proteins. These results showed the importance of a functional QS system in R. rubrum.
Project description:This SuperSeries is composed of the following subset Series: GSE14239: MESSAGE 2 space experiment with Rhodospirillum rubrum S1H GSE14241: BASE-A space experiment with Rhodospirillum rubrum S1H Refer to individual Series
Project description:Aiming at the development of a micropollutant biosensor in the frame of the Micro-Ecological Life Support System Alternative (MELiSSA), a pilot study was initiated to identify triclosan (TCS)-responsive biomarker genes in the MELiSSA carbon-mineralizing microorganism, Rhodospirillum rubrum S1H. TCS is a biocide, commonly found in human excrements and is considered an emerging pollutant in wastewater and the environment. Chronic exposure to MELiSSA-relevant concentrations (≥25 µg L-1) TCS caused a significant extension of the lag phase without affecting the growth rate. Analytical determination gave no indication of TCS biodegradation during the growth experiment and flow cytometric viability analyses revealed that TCS is only slightly toxic to R. rubrum. Through microarray analyses, the genetic mechanisms supporting the reversibility of TCS-induced inhibition were scrutinized. Hence, an extremely TCS-responsive cluster of four small adjacent genes was revealed, with up to 34-fold induction at 25 µg L-1. These four genes, for which the name micropollutant-upregulated factor (muf) was proposed; appear to be unique to R. rubrum and are shown for the first time to be involved in the response to stress. Moreover, numerous other systems that are associated with the proton-motive force were shown to be responsive to TCS, but never as highly upregulated as the muf genes. Hence, R. rubrum induced the phage shock protein operon (pspABC), numerous major facilitator efflux systems, cell envelope consolidation mechanisms, oxidative stress response, beta-oxidation, and carbonic anhydrase; while downregulating bacterial conjugation- and carboxysome synthesis genes. The muf genes and three efflux-related genes showed most potential as low-dose biomarkers. The two microarray experiments (10 and 25 µg l-1 Triclosan) were all performed in biological triplicate and containing three (in-slide) technical repeats. For all conditions, the Triclosan exposed sample (Cy5) was compared with the non-exposed solvent control (Cy3) to identify those genes that were differentially expressed upon Triclosan exposure.
Project description:Aiming at the development of a micropollutant biosensor in the frame of the Micro-Ecological Life Support System Alternative (MELiSSA), a pilot study was initiated to identify triclosan (TCS)-responsive biomarker genes in the MELiSSA carbon-mineralizing microorganism, Rhodospirillum rubrum S1H. TCS is a biocide, commonly found in human excrements and is considered an emerging pollutant in wastewater and the environment. Chronic exposure to MELiSSA-relevant concentrations (≥25 µg L-1) TCS caused a significant extension of the lag phase without affecting the growth rate. Analytical determination gave no indication of TCS biodegradation during the growth experiment and flow cytometric viability analyses revealed that TCS is only slightly toxic to R. rubrum. Through microarray analyses, the genetic mechanisms supporting the reversibility of TCS-induced inhibition were scrutinized. Hence, an extremely TCS-responsive cluster of four small adjacent genes was revealed, with up to 34-fold induction at 25 µg L-1. These four genes, for which the name micropollutant-upregulated factor (muf) was proposed; appear to be unique to R. rubrum and are shown for the first time to be involved in the response to stress. Moreover, numerous other systems that are associated with the proton-motive force were shown to be responsive to TCS, but never as highly upregulated as the muf genes. Hence, R. rubrum induced the phage shock protein operon (pspABC), numerous major facilitator efflux systems, cell envelope consolidation mechanisms, oxidative stress response, beta-oxidation, and carbonic anhydrase; while downregulating bacterial conjugation- and carboxysome synthesis genes. The muf genes and three efflux-related genes showed most potential as low-dose biomarkers.