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SRX13191397: GSM5699191: Free-living cells with F. serratus rep 2; Zobellia galactanivorans; RNA-Seq
1 ILLUMINA (NextSeq 2000) run: 26.2M spots, 2G bases, 809.8Mb downloads

External Id: GSM5699191_r1
Submitted by: Marine Glycobiology group, UMR8227, CNRS-UPMC, Station Biologique de Roscoff
Study: Unraveling the metabolic mechanisms of pioneer bacteria during the utilization of fresh macroalgae
show Abstracthide Abstract
We cultivated the flavobacterium Zobellia galactanivorans DsijT with fresh brown macroalgae with distinct chemical compositions. Its capacity to use macroalgae as the sole carbon source via the secretion of extracellular enzymes, leading to extensive tissue damages, highlights a sharing pioneer degrader behavior. RNA-seq transcriptome analysis revealed a metabolic shift toward the utilization of brown algal polysaccharides during tissue degradation. A subset of genes was specifically induced in cells grown with intact algae compared to purified polysaccharides. It notably includes genes involved in protection against oxidative burst, type IX secretion system proteins and novel uncharacterized Polysaccharides Utilization Loci (PULs). Comparative growth experiments and genomics between Zobellia members brought out putative genetic determinants of the pioneer behavior of Z. galactanivorans, whose in vitro role could be further characterized. This work constitutes the first investigation of the metabolic mechanisms of bacteria mediating fresh macroalgae breakdown, and will help unravel the role of marine microbes in the fate of macroalgal biomass. Overall design: RNAseq profiles of Zobellia galactanivorans DsijT when grown with different carbon sources. Profiles of free-living cells were assessed with Laminaria digitata, Fucus serratus, Ascophyllum nodosum, Maltose, Alginate or Fucose-containing sulfated polysaccharides (FCSP) as the sole carbon source. Profiles of algae-attached cells were assessed with Laminaria digitata and Fucus serratus as the sole carbon source. Experiments were done in triplicates for each condition (except for free-living cells with F. serratus and algae-attached cells where n=2). 21 samples in total. Maltose condition was used as a control.
Sample: Free-living cells with F. serratus rep 2
SAMN23384253 • SRS11119489 • All experiments • All runs
Library:
Name: GSM5699191
Instrument: NextSeq 2000
Strategy: RNA-Seq
Source: TRANSCRIPTOMIC
Selection: cDNA
Layout: PAIRED
Construction protocol: Free-living bacterial cells were lysed by adding 400 μl of lysis buffer (4 M guanidine thiocyanate, 25 mM sodium acetate pH 5.2, 5 g.l-1 N-laurylsarcosinate) and 500 μl of phenol and incubated 5 min at 65 °C. RNA was extracted using phenol-chloroform. Extracts were treated 1 h at 37 °C with 2 units of Turbo DNAse and purified on mini-columns NucleoSpin RNA Clean-up following the manufacturer's instructions. RNA was eluted in 50 μl of nuclease-free water. RNA from algae-attached bacteria was extracted as follows. Two algal pieces were immersed in killing buffer, vortexed and placed 7 min in an ultrasonic bath to detach bacteria from the algal surface. Algae were removed and cell pellets resuspended in lysis buffer. RNA extraction and DNAse treatment were performed as described above for the free-living bacteria. DNAse were inactivated using DNAse inactivation reagent following the manufacturer instructions. Librairies were performed by the Plateforme de Séquençage I2BC (UMR9198, CNRS, Gif-sur-Yvette) on a NextSeq instrument (Illumina) using the NextSeq 500/550 High Output Kit v2 (75 cycles) which included a Ribo-Zero ribosomal RNA depletion step.
Runs: 1 run, 26.2M spots, 2G bases, 809.8Mb
Run# of Spots# of BasesSizePublished
SRR1700118926,236,0642G809.8Mb2022-07-27

ID:
18051083

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