Project description:Diel cycle is of enormous biological importance in that it imposes temporal structure on ecosystem productivity. In the world oceans, microorganisms form complex communities that carry out about half of photosynthesis and the bulk of life-sustaining nutrient cycling. Within these natural microbial assemblages, photoautotrophs, such as Cyanobacteria, display diel rhythmicity in gene expression. To what extent autotrophs and heterotrophs are impacted by light and dark oscillations and how this collectively influences community structure and functionality remains poorly documented. In this study, we compared eight day/night metaproteome profiles of Cyanobacteria and both free-living and attached bacterial fractions from picoplanktonic communities sampled over two consecutive days from the surface north-west Mediterranean Sea. Our results showed similar taxonomic structure in both free-living and particle-attached bacteria, dominated by Alphaproteobacteria and Gammaproteobacteria. Temporal rhythmicity in protein expression was observed in both Synechococcales and Rhodobacterales in light-dependent processes such as photosynthesis or UV-stress response. Other biological processes, such as phosphorus or amino acid metabolisms, were also found to cycle in phototrophs. In contrast, proteins from the ubiquitous Pelagibacterales remained stable independently of the day/night oscillations. This work integrated for the first time diel comparative metaproteomics on both free-living and particle attached bacterial fractions in coastal oligotrophic environment. Our findings demonstrated a taxa-specific response to diel cycle with a more controlled protein regulation for phototrophs. This study provided additional evidences that timekeeping mechanisms might be widespread among Bacteria, broadening our knowledge on diel microbial assemblage dynamics.
2020-03-06 | PXD017373 | iProX
Project description:particle-attached and free-living bacteria kongsfjorden
| PRJNA488164 | ENA
Project description:The free-living and particle-attached bacterial communities in seawater of Yangtze River Estuary
Project description:Microarrays are useful tools for detecting and quantifying specific functional and phylogenetic genes in natural microbial communities. In order to track uncultivated microbial genotypes and their close relatives in an environmental context, we designed and implemented a “genome proxy” microarray that targets microbial genome fragments recovered directly from the environment. Fragments consisted of sequenced clones from large-insert genomic libraries from microbial communities in Monterey Bay, the Hawaii Ocean Time-series station ALOHA, and Antarctic coastal waters. In a prototype array, we designed probe sets to thirteen of the sequenced genome fragments and to genomic regions of the cultivated cyanobacterium Prochlorococcus MED4. Each probe set consisted of multiple 70-mers, each targeting an individual ORF, and distributed along each ~40-160kbp contiguous genomic region. The targeted organisms or clones, and close relatives, were hybridized to the array both as pure DNA mixtures and as additions of cells to a background of coastal seawater. This prototype array correctly identified the presence or absence of the target organisms and their relatives in laboratory mixes, with negligible cross-hybridization to organisms having ≤~75% genomic identity. In addition, the array correctly identified target cells added to a background of environmental DNA, with a limit of detection of ~0.1% of the community, corresponding to ~10^3 cells/ml in these samples. Signal correlated to cell concentration with an R2 of 1.0 across six orders of magnitude. In addition the array could track a related strain (at 86% genomic identity to that targeted) with a linearity of R2=0.9999 and a limit of detection of ~1% of the community. Closely related genotypes were distinguishable by differing hybridization patterns across each probe set. This array’s multiple-probe, “genome-proxy” approach and consequent ability to track both target genotypes and their close relatives is important for the array’s environmental application given the recent discoveries of considerable intra-population diversity within marine microbial communities. Keywords: target addition experiment, proof-of-concept for GPL6012