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. 2010 Jan;12(1):74-82.
doi: 10.1111/j.1462-2920.2009.02043.x. Epub 2009 Aug 27.

Contrasting ability to take up leucine and thymidine among freshwater bacterial groups: implications for bacterial production measurements

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Free PMC article

Contrasting ability to take up leucine and thymidine among freshwater bacterial groups: implications for bacterial production measurements

María Teresa Pérez et al. Environ Microbiol. 2010 Jan.
Free PMC article

Abstract

We examined the ability of different freshwater bacterial groups to take up leucine and thymidine in two lakes. Utilization of both substrates by freshwater bacteria was examined at the community level by looking at bulk incorporation rates and at the single-cell level by combining fluorescent in situ hybridization and signal amplification by catalysed reporter deposition with microautoradiography. Our results showed that leucine was taken up by 70-80% of Bacteria-positive cells, whereas only 15-43% of Bacteria-positive cells were able to take up thymidine. When a saturating substrate concentration in combination with a short incubation was used, 80-90% of Betaproteobacteria and 67-79% of Actinobacteria were positive for leucine uptake, whereas thymidine was taken up by < 10% of Betaproteobacteria and by < 1% of the R-BT subgroup that dominated this bacterial group. Bacterial abundance was a good predictor of the relative contribution of bacterial groups to leucine uptake, whereas when thymidine was used Actinobacteria represented the large majority (> 80%) of the cells taking up this substrate. Increasing the substrate concentration to 100 nM did not affect the percentage of R-BT cells taking up leucine (> 90% even at low concentrations), but moderately increased the fraction of thymidine-positive R-BT cells to a maximum of 35% of the hybridized cells. Our results show that even at very high concentrations, thymidine is not taken up by all, otherwise active, bacterial cells.

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Figures

Fig. 1
Fig. 1
Percentage of cells active for Leu (solid bars) and TdR (open bars) uptake within the most abundant bacterial groups: EUB (Bacteria; A), BETA (Betaproteobacteria; B), R-BT (subgroup of Betaproteobacteria; C), HGC (Actinobacteria; D), ALFA (Alphaproteobacteria; E) and CF (Cytophaga-like cells; F). GKS 1 and GKS 8.5 represent samples collected in September 2006 in GKS at 1 and 8.5 m depth respectively. SOS 06 and SOS 07 represent samples collected in SOS at 1 m depth in September 2006 and August 2007 respectively. Values are mean of three replicates ± 1 SD.
Fig. 2
Fig. 2
Relative contribution of Betaproteobacteria (BETA), Actinobacteria (HGC), Cytophaga-like bacteria (CF) and Alphaproteobacteria (ALFA) to Leu (A) or TdR (B) uptake plotted against their relative abundance among Bacteria. Values are mean of three replicates ± 1 SD. The diagonal line represents a 1:1 relationship.
Fig. 3
Fig. 3
Fractions of Bacteria (probe EUB I-III), Betaproteobacteria (probe BET42a) and R-BT cells (probe R-BT065) positive for Leu (left) and TdR (right) uptake at different substrate concentrations. Dots and lines show the corresponding bulk incorporation rates of Leu and TdR by the whole bacterial assemblage.

References

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