Heterotrophic Prokaryotes Communities Research Articles

Effects of Phosphorus Limitation on the Bioavailability of DOM Released by Marine Heterotrophic Prokaryotes.

Heterotrophic prokaryotes (HP) contribute largely to dissolved organic matter (DOM) processing in the ocean, but they also release diverse organic substances. The bioavailability of DOM released by HP under varying environmental conditions has not been fully elucidated. In this study, we investigated the bioavailability of DOM released by a single bacterial strain (Sphingopyxis alaskensis) and 2 natural HP communities grown under P-replete and P-limited conditions. The released DOM (HP-DOM) was used as a substrate for natural HP communities at a coastal site in the Northwestern Mediterranean Sea. We followed changes in HP growth, enzymatic activity, diversity, and community composition together with the consumption of HP-DOM fluorescence (FDOM). HP-DOM produced under P-replete and P-limited conditions promoted significant growth in all incubations. No clear differences in HP-DOM lability released under P-repletion and P-limitation were evidenced based on the HP growth, and P-limitation was not demonstrated to decrease HP-DOM lability. However, HP-DOM supported the growth of diverse HP communities, and P-driven differences in HP-DOM quality were selected for different indicator taxa in the degrading communities. The humic-like fluorescence, commonly considered recalcitrant, was consumed during the incubations when this peak was initially dominating the FDOM pool, and this consumption coincided with higher alkaline phosphatase activity. Taken together, our findings emphasize that HP-DOM lability is dependent on both DOM quality, which is shaped by P availability, and the composition of the consumer community.

Selective enrichment on a wide polysaccharide spectrum allowed isolation of novel metabolic and taxonomic groups of haloarchaea from hypersaline lakes.

Extremely halophilic archaea (haloarchaea) of the class Halobacteria is a dominant group of aerobic heterotrophic prokaryotic communities in salt-saturated habitats, such as salt lakes and solar salterns. Most of the pure cultures of haloarchaea were enriched, isolated, and cultivated on rich soluble substrates such as amino acids, peptides or simple sugars. So far, the evidences on the capability of haloarchaea to use different polysaccharides as growth substrates remained scarce. However, it is becoming increasingly obvious that these archaea can also actively participate in mineralization of complex biopolymers, in particular cellulose and chitin-two dominant biomass polysaccharides on the planet. Here we used an array of commercially available homo- and heteropolysaccharides to enrich hydrolytic haloarchaea from hypersaline salt lakes with neutral pH and from alkaline soda lakes. This resulted in isolation of a range of halo- and natrono-archaea, respectively, belonging to already described taxa as well as several new genus-level lineages. In some cases, the isolates enriched with different polysaccharides happened to be closely related, thus representing generalistic ecotype, while the others were narrow specialists. In general, soda lakes yielded a broader range of polysaccharide-utilizing specialists in comparison to neutral salt lakes. The results demonstrated a significant diversity of halo(natrono)archaea with a previously unrecognized potential for utilization of a broad range of natural polysaccharides in hypersaline habitats.

Phosphorus limitation determines the quality of dissolved organic matter released by marine heterotrophic prokaryotes

AbstractWe determined phosphorus (P) limitation effect on the quantity and quality of dissolved organic matter (DOM) released by heterotrophic prokaryotes (HP). We grew two single‐bacterial strains from different lifestyles, the copiotrophic Photobacterium angustum and the oligotrophic Sphingopyxis alaskensis, and natural HP communities collected in fall and spring from the Mediterranean Sea, on glucose under two treatments: P‐replete versus P‐limiting. DOM release by HP comprised up to 30% of the initial carbon provided for growth. P‐availability influenced carbon allocation to different cellular processes (respiration vs. growth), but did not significantly affect the net quantity of DOM released by HP. However, using fluorescence spectroscopy, we demonstrated an effect of P‐limitation on DOM quality, with a predominance of humic‐like compounds under P‐limitation but protein‐like compounds under P‐repletion. Our results suggest that P‐limitation could determine the fate of HP‐derived DOM in the ocean, thus affecting the microbial carbon pump.

Prokaryotic diversity and activity in contrasting productivity regimes in late summer in the Kerguelen region (Southern Ocean)

Natural iron (Fe) fertilization sustains phytoplankton blooms above the Kerguelen plateau (Indian sector of the Southern Ocean) within otherwise low productive off-plateau waters. In early spring and summer, these diatom-dominated blooms are associated with distinct heterotrophic prokaryotic communities, but whether a structuring effect extends to the post-bloom period has thus far not been investigated. To address this question, we carried out a detailed study of the prokaryotic community composition in the region of Kerguelen Island during late Austral summer (18 February to 27 March 2018; MOBYDICK project). Concentrations of chlorophyll a were seasonally low above the plateau (0.27–0.58 μg Chl a L −1 ) and in a similar range to those at the 3 off-plateau sites investigated (0.14–0.34 μg Chl a L −1 ), but we observed an accumulation of dissolved organic carbon and the build-up of heterotrophic prokaryotic biomass in Kerguelen plateau waters. Illumina sequencing of the 16S rRNA gene revealed that the total (DNA-based) and potentially active (RNA-based) prokaryotic communities were structured according to on- and off-plateau sites in the wind-mixed surface layer, in both the free-living (<0.8 μm size fraction) and particle-attached (>0.8 μm size fraction) fractions. The Amplicon Sequence Variants (ASV) with significantly higher relative abundances in on-plateau surface waters as compared to off-plateau waters belonged to Halieaceae OM60 group, several Flavobacteriaceae , such as the NS5 marine group, Aurantivirga and Ulvibacter , Rhodobacteraceae Loktanella , Saprospiraceae , and the Cryomorphaceae NS10 marine group. ASVs with higher relative abundances in off-plateau waters belonged to the Flavobacteriaceae Formosa , the Rhodobacteraceae Planktomarina and the SAR11 clade. We discuss the potential abiotic and biotic drivers of community composition in late Austral summer and the ecological roles of abundant prokaryotic taxa in Kerguelen plateau waters.

Dissolved organic matter released by two marine heterotrophic bacterial strains and its bioavailability for natural prokaryotic communities.

Marine heterotrophic prokaryotes (HP) play a key role in organic matter processing in the ocean; however, the view of HP as dissolved organic matter (DOM) sources remains underexplored. In this study, we quantified and optically characterized the DOM produced by two single marine bacterial strains. We then tested the availability of these DOM sources to in situ Mediterranean Sea HP communities. Two bacterial strains were used: Photobacterium angustum (a copiotrophic gammaproteobacterium) and Sphingopyxis alaskensis (an oligotrophic alphaproteobacterium). When cultivated on glucose as the sole carbon source, the two strains released from 7% to 23% of initial glucose as bacterial derived DOM (B-DOM), the quality of which (as enrichment in humic or protein-like substances) differed between strains. B-DOM induced significant growth and carbon consumption of natural HP communities, suggesting that it was partly labile. However, B-DOM consistently promoted lower prokaryotic growth efficiencies than in situ DOM. In addition, B-DOM changed HP exoenzymatic activities, enhancing aminopeptidase activity when degrading P. angustum DOM, and alkaline phosphatase activity when using S. alaskensis DOM, and promoted differences in HP diversity and composition. DOM produced by HP affects in situ prokaryotic metabolism and diversity, thus changing the pathways for DOM cycling (e.g. respiration over biomass production) in the ocean.

Significance of Viral Activity for Regulating Heterotrophic Prokaryote Community Dynamics along a Meridional Gradient of Stratification in the Northeast Atlantic Ocean.

How microbial populations interact influences the availability and flux of organic carbon in the ocean. Understanding how these interactions vary over broad spatial scales is therefore a fundamental aim of microbial oceanography. In this study, we assessed variations in the abundances, production, virus and grazing induced mortality of heterotrophic prokaryotes during summer along a meridional gradient in stratification in the North Atlantic Ocean. Heterotrophic prokaryote abundance and activity varied with phytoplankton biomass, while the relative distribution of prokaryotic subpopulations (ratio of high nucleic acid fluorescent (HNA) and low nucleic acid fluorescent (LNA) cells) was significantly correlated to phytoplankton mortality mode (i.e., viral lysis to grazing rate ratio). Virus-mediate morality was the primary loss process regulating the heterotrophic prokaryotic communities (average 55% of the total mortality), which may be attributed to the strong top-down regulation of the bacterivorous protozoans. Host availability, encounter rate, and HNA:LNA were important factors regulating viral dynamics. Conversely, the abundance and activity of bacterivorous protozoans were largely regulated by temperature and turbulence. The ratio of total microbial mediated mortality to total available prokaryote carbon reveals that over the latitudinal gradient the heterotrophic prokaryote community gradually moved from a near steady state system regulated by high turnover in subtropical region to net heterotrophic production in the temperate region.

Diel dynamics and coupling of heterotrophic prokaryotes and dissolved organic matter in epipelagic and mesopelagic waters of the central Red Sea.

The ecological status of an ecosystem can be approached by the taxa present but also by the size of individual organisms. In aquatic ecosystems, flow cytometry (FC) allows to study the individual size spectra and broad community composition through the evaluation of cytometric categories. The Red Sea represents a warm oligotrophic environment with a strong diel signal of vertically migrating mesopelagic fish, which feed at night at the surface and release dissolved organic carbon (DOC) at depth during day-time. However, knowledge about how these conditions affect the dynamics of heterotrophic prokaryotes (HP) and their coupling with DOC is lacking. Here, we analyzed a high frequency sampling over 24 h to identify the community structure and compositional changes of HP in the epipelagic and mesopelagic layers of the central Red Sea. Our results show marked vertical and diel changes in HP communities in both layers. Specifically, the relative contribution of high nucleic acid content cells was remarkably linked to changes in DOC concentration and properties. The patterns observed were likely associated to the diel vertical migration of mesopelagic fish. These findings reveal that the structure of microbial communities in warm oligotrophic environments may be more dynamic than previously thought.

Microbial weeds in hypersaline habitats: the enigma of the weed-like Haloferax mediterranei.

Heterotrophic prokaryotic communities that inhabit saltern crystallizer ponds are typically dominated by two species, the archaeon Haloquadratum walsbyi and the bacterium Salinibacter ruber, regardless of location. These organisms behave as 'microbial weeds' as defined by Cray etal. (Microb Biotechnol 6: 453-492, 2013) that possess the biological traits required to dominate the microbiology of these open habitats. Here, we discuss the enigma of the less abundant Haloferax mediterranei, an archaeon that grows faster than any other, comparable extreme halophile. It has a wide window for salt tolerance, can grow on simple as well as on complex substrates and degrade polymeric substances, has different modes of anaerobic growth, can accumulate storage polymers, produces gas vesicles, and excretes halocins capable of killing other Archaea. Therefore, Hfx.mediterranei is apparently more qualified as a 'microbial weed' than Haloquadratum and Salinibacter. However, the former differs because it produces carotenoid pigments only in the lower salinity range and lacks energy-generating retinal-based, light-driven ion pumps such as bacteriorhodopsin and halorhodopsin. We discuss these observations in relation to microbial weed biology in, and the open-habitat ecology of, hypersaline systems.

Seasonal patterns of viral, microbial and planktonic communities in Sidi Salem: a freshwater reservoir (North of Tunisia)

We investigated the distribution and dynamics of viruses, prokaryotes and small eukaryotic phytoplankton in Sidi Salem freshwater reservoir (Northern Tunisia). Samples were collected from the deepest station at different depths throughout the water column for 2 years (February 2009 to January 2011). The reservoir was characterized by seasonal alternations of thermal stratification and homothermy. Among the different microbial communities counted using flow cytometry (FCM), picocyanobacteria constituted an important autotrophic component since they were always present and their highest concentration reached 3.02 and 2.65×105 cells.mL−1 in March 2009 and June 2010, respectively. The heterotrophic prokaryotic communities (represented mainly by bacteria) were characterized by a clear separation between two subgroups referred to as high-DNA and low-DNA content populations, and the highest concentrations of heterotrophic bacteria (i.e., 3.8×107 cells.mL−1) were recorded in spring 2009. Several viral groups referred to as virus-like particles (VLP) groups 1, 2 and 3 could also be discriminated using FCM. VLP1 and VLP2 displayed a significant correlation with the heterotrophic bacteria (r=0.80 and 0.78, P<0.001) but seem to be independent from picocyanobacteria and/or chlorophyll a, suggesting these VLPs were mainly bacteriophages. At last, the virus to prokaryotic ratio could be high, especially in summer (mean=22, max=487), suggesting a strong coupling between bacteria and viruses, at least at certain periods of the year.

Making a living while starving in the dark: metagenomic insights into the energy dynamics of a carbonate cave

Carbonate caves represent subterranean ecosystems that are largely devoid of phototrophic primary production. In semiarid and arid regions, allochthonous organic carbon inputs entering caves with vadose-zone drip water are minimal, creating highly oligotrophic conditions; however, past research indicates that carbonate speleothem surfaces in these caves support diverse, predominantly heterotrophic prokaryotic communities. The current study applied a metagenomic approach to elucidate the community structure and potential energy dynamics of microbial communities, colonizing speleothem surfaces in Kartchner Caverns, a carbonate cave in semiarid, southeastern Arizona, USA. Manual inspection of a speleothem metagenome revealed a community genetically adapted to low-nutrient conditions with indications that a nitrogen-based primary production strategy is probable, including contributions from both Archaea and Bacteria. Genes for all six known CO2-fixation pathways were detected in the metagenome and RuBisCo genes representative of the Calvin-Benson-Bassham cycle were over-represented in Kartchner speleothem metagenomes relative to bulk soil, rhizosphere soil and deep-ocean communities. Intriguingly, quantitative PCR found Archaea to be significantly more abundant in the cave communities than in soils above the cave. MEtaGenome ANalyzer (MEGAN) analysis of speleothem metagenome sequence reads found Thaumarchaeota to be the third most abundant phylum in the community, and identified taxonomic associations to this phylum for indicator genes representative of multiple CO2-fixation pathways. The results revealed that this oligotrophic subterranean environment supports a unique chemoautotrophic microbial community with potentially novel nutrient cycling strategies. These strategies may provide key insights into other ecosystems dominated by oligotrophy, including aphotic subsurface soils or aquifers and photic systems such as arid deserts.

Lack of P-limitation of phytoplankton and heterotrophic prokaryotes in surface waters of three anticyclonic eddies in the stratified Mediterranean Sea

Abstract. We investigated the identity of the limiting nutrient of the pelagic microbial food web in the Mediterranean Sea using nutrient manipulated microcosms during summer 2008. Experiments were carried out with surface waters at the center of anticyclonic eddies in the Western Basin, the Ionian Basin, and the Levantine Basin. In situ, the ratio of N to P was always higher in both dissolved and particulate organic fractions compared to the Redfield ratio, suggesting a relative P-starvation. In each experiment, four different treatments in triplicates (addition of ammonium, phosphate, a combination of both, and the unamended control) were employed and chemical and biological parameters monitored throughout a 3–4 day incubation. Temporal changes of turnover time of phosphate and ATP, and alkaline phosphatase activity during the incubation suggested that the phytoplankton and heterotrophic prokaryotes (Hprok) communities were not P-limited at the sites. Furthermore, statistical comparison among treatments at the end of the incubation did not support a hypothesis of P-limitation at the three study sites. In contrast, primary production was consistently limited by N, and Hprok growth was not limited by N nor P in the Western Basin, but N-limited in the Ionian Basin, and N and P co-limited in the Levantine Basin. Our results demonstrated the gap between biogeochemical features (an apparent P-starved status) and biological responses (no apparent P-limitation). We question the general notion that Mediterranean surface waters are limited by P alone during the stratified period.