Exploring the potential of a Functional Gene Array (FGA) for fluvial biofilms
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ABSTRACT:
PROVIDER: PRJNA128659 | ENA |
REPOSITORIES: ENA
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Project description:Microarrays offer a comprehensive tool for a system-wide analysis of cell functioning based on the simultaneous detection of the activity of thousands of genes. While transcriptome analyses are usually related with the study of gene expression and regulation within single species in laboratory experiments or environmental samples, in the present study, the possibility of studying gene expression in river biofilm communities was explored. To this aim a Functional Gene array (FGA), based on consensus sequences from genes of key physiological processes, was designed including 83 functional genes chosen to reflect several essential biochemical pathways and specific stress response pathways. Probes of these genes were designed from consensus sequences from up to 6 microalgal species (diatoms and chlorophytes). Furthermore, species specific probes were included resulting in 1556 unique oligonucleotide probes for 83 different genes. RNA extracted from Chlamydomonas reinhardtii, Scenedesmus vacuolatus and multi-species biofilms was then hybridized to oligonucleotide arrays at different hybridization temperatures (55, 60 and 65ºC). Signal intensity was affected by sequence divergence but results showed that hybridisation temperature of 55ºC allowed a good compromise between cross-hybridization and specificity. Due to the limited number of probes and available sequence information the designed FGAs is at present particularly useful for the comparison of similar biofilms exposed to different conditions in laboratory experimental studies.
2012-12-31 | GSE22496 | GEO
Project description:Microarrays offer a comprehensive tool for a system-wide analysis of cell functioning based on the simultaneous detection of the activity of thousands of genes. While transcriptome analyses are usually related with the study of gene expression and regulation within single species in laboratory experiments or environmental samples, in the present study, the possibility of studying gene expression in river biofilm communities was explored. To this aim a Functional Gene array (FGA), based on consensus sequences from genes of key physiological processes, was designed including 83 functional genes chosen to reflect several essential biochemical pathways and specific stress response pathways. Probes of these genes were designed from consensus sequences from up to 6 microalgal species (diatoms and chlorophytes). Furthermore, species specific probes were included resulting in 1556 unique oligonucleotide probes for 83 different genes. RNA extracted from Chlamydomonas reinhardtii, Scenedesmus vacuolatus and multi-species biofilms was then hybridized to oligonucleotide arrays at different hybridization temperatures (55, 60 and 65ºC). Signal intensity was affected by sequence divergence but results showed that hybridisation temperature of 55ºC allowed a good compromise between cross-hybridization and specificity. Due to the limited number of probes and available sequence information the designed FGAs is at present particularly useful for the comparison of similar biofilms exposed to different conditions in laboratory experimental studies. Nine samples were used, three were extracted from biofilm communities and hybridized at 55, 60 and 65 ºC, respectively; three were extracted from Scenedesmus vacuolatus and hybridized at 55, 60 and 65 ºC, respectively; and the three last ones were extracted from Chlamydomonas reinhardtii and hybridized at 55, 60 and 65 ºC
2012-12-31 | E-GEOD-22496 | biostudies-arrayexpress
Project description:Microtoming Coupled with Microarray Analysis to Evaluate Potential Differences in the Metabolic Status of Geobacter sulfurreducens at Different Depths in Anode Biofilms Differences in the Metabolic Status of Geobacter sulfurreducens at Different Depths in A Current Producing Biofilm Further insight into the metabolic status of cells within anode biofilms is essential for understanding the functioning of microbial fuel cells and developing strategies to optimize their power output. In order to further compare the metabolic status of cells growing close to the anode versus cells in the outer portion of the anode biofilm, mature anode biofilms were treated to stop turnover over of mRNA and then encased in resin which was sectioned into 100 nm shavings with a diamond knife and pooled into inner (0-20 µm from anode surface) and outer (30-60 µm) fractions. Whole genome DNA microarray analysis of RNA extracted from the shavings revealed that, at a 2-fold lower threshold, there were 146 genes that had significant (p<0.05), differences in transcript abundance between the inner and outer portions of the biofilm. Only 1 gene, GSU0093, a hypothetical ABC transporter, had significantly higher transcript abundances in the outer biofilm. Genes with lower transcript abundance in the outer biofilm included genes for ribosomal proteins and NADH dehydrogenase, suggesting that cells in the outer biofilm had lower metabolic rates. However, the differences in transcript abundance were relatively low (<3-fold) and the outer biofilm did not have significantly lower expression of the genes for TCA cycle enzymes which previous studies have demonstrated are sensitive indicators of changes in rates of metabolism in G. sulfurreducens. There also was no significant difference in the transcript levels for outer-surface cell components thought to be important in electron transfer in anode biofilms. Lower expression of genes involved in stress responses in the outer biofilm may reflect the development of low pH near the surface of the anode. The results of the metabolic staining and gene expression studies suggest that cells throughout the biofilm are metabolically active and can potentially contribute to current production. The microtoming/microarray strategy described here may be useful for evaluating gene expression with depth in a diversity of microbial biofilms. Three biological replicates were hybridized in triplicate on a coustom affimetrix tilling array using prokaryotic protocol (p69Affy, p75 Adobe) for labeling, hybridization and scanning.
2010-06-21 | E-GEOD-17591 | biostudies-arrayexpress
Project description:Microtoming Coupled with Microarray Analysis to Evaluate Potential Differences in the Metabolic Status of Geobacter sulfurreducens at Different Depths in Anode Biofilms Differences in the Metabolic Status of Geobacter sulfurreducens at Different Depths in A Current Producing Biofilm Further insight into the metabolic status of cells within anode biofilms is essential for understanding the functioning of microbial fuel cells and developing strategies to optimize their power output. In order to further compare the metabolic status of cells growing close to the anode versus cells in the outer portion of the anode biofilm, mature anode biofilms were treated to stop turnover over of mRNA and then encased in resin which was sectioned into 100 nm shavings with a diamond knife and pooled into inner (0-20 µm from anode surface) and outer (30-60 µm) fractions. Whole genome DNA microarray analysis of RNA extracted from the shavings revealed that, at a 2-fold lower threshold, there were 146 genes that had significant (p<0.05), differences in transcript abundance between the inner and outer portions of the biofilm. Only 1 gene, GSU0093, a hypothetical ABC transporter, had significantly higher transcript abundances in the outer biofilm. Genes with lower transcript abundance in the outer biofilm included genes for ribosomal proteins and NADH dehydrogenase, suggesting that cells in the outer biofilm had lower metabolic rates. However, the differences in transcript abundance were relatively low (<3-fold) and the outer biofilm did not have significantly lower expression of the genes for TCA cycle enzymes which previous studies have demonstrated are sensitive indicators of changes in rates of metabolism in G. sulfurreducens. There also was no significant difference in the transcript levels for outer-surface cell components thought to be important in electron transfer in anode biofilms. Lower expression of genes involved in stress responses in the outer biofilm may reflect the development of low pH near the surface of the anode. The results of the metabolic staining and gene expression studies suggest that cells throughout the biofilm are metabolically active and can potentially contribute to current production. The microtoming/microarray strategy described here may be useful for evaluating gene expression with depth in a diversity of microbial biofilms.
2010-06-22 | GSE17591 | GEO
Project description:Biofilms on microplastics – composition, genetic potential, and function examined by meta-proteogenomics
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Project description:In nature, bacteria reside in biofilms - multicellular differentiated communities held together by extracellular matrix. In this work, we identified a novel subpopulation essential for biofilm formation – mineral-forming cells in Bacillus subtilis biofilms. This subpopulation contains an intracellular calcium-accumulating niche, in which the formation of a calcium carbonate mineral is initiated. As the biofilm colony develops, this mineral grows in a controlled manner, forming a functional macrostructure that serves the entire community. Consistently, biofilm development is prevented by inhibition of calcium uptake. Taken together, our results provide a clear demonstration of the orchestrated production of calcite exoskeleton, critical to morphogenesis in simple prokaryotes. We expect future research exploring this newly discovered process to shed further light on mechanisms of bacterial development.
2022-03-24 | GSE199052 | GEO