Strong seasonal differences of bacterial polysaccharide utilization in the North Sea over an annual cycle

Marine heterotrophic bacteria contribute considerably to global carbon cycling, in part by utilizing phytoplankton-derived polysaccharides. The patterns and rates of two different polysaccharide utilization modes – extracellular hydrolysis and selfish uptake – have previously been found to change during spring phytoplankton bloom events. Here we investigated seasonal changes in bacterial utilization of three polysaccharides, laminarin, xylan and chondroitin sulfate. Strong seasonal differences were apparent in mode and speed of polysaccharide utilization, as well as in bacterial community compositions. Compared to the winter month of February, during the spring bloom in May, polysaccharide utilization was detected earlier in the incubations and a higher portion of all bacteria took up laminarin selfishly. Highest polysaccharide utilization was measured in June and September, mediated by bacterial communities that were significantly different from spring assemblages. Extensive selfish laminarin uptake, for example, was detectible within a few hours in June, while extracellular hydrolysis of chondroitin was dominant in September. In addition to the well-known Bacteroidota and Gammaproteobacteria clades, the numerically minor verrucomicrobial clade Pedosphaeraceae could be identified as a rapid laminarin utilizer. In summary, polysaccharide utilization proved highly variable over the seasons, both in mode and speed, and also by the bacterial clades involved.     

An Inhibitor-Based Method To Measure Initial Decomposition of Naturally Occurring Polysaccharides in Sediments

A method that can be used to measure the initial decomposition rates of polysaccharides in sediment samples was developed. It uses toluene to specifically inhibit microbial uptake of carbohydrates without affecting extracellular hydrolysis of polysaccharides. Accumulating carbohydrates were determined by high-performance liquid chromatography. Field-sampled litter from the common reed (Phragmites australis), which contains cellulose and arabinoxylan as its main polysaccharides, was used as a model system. Toluene concentrations of between 1 and 10% resulted in the accumulation of similar amounts of monomeric carbohydrates, which was linear over time for most neutral sugars. Toluene (3%) did not have an effect on extracellular enzyme activities, and microbial sugar uptake was completely inhibited, as demonstrated with (sup14)C-labelled xylose and glucose. Experiments with enhancement cultures and fixed reed litter suggested that enzymatic hydrolysis of polysaccharides in reed litter was the main source of glucose, xylose, arabinose, and galactose accumulation. In contrast, the accumulation of high amounts of the alditols mannitol and glucitol was probably caused by lysis of the microbial population in toluene-treated reed litter. Glucose accumulated at rates of 1.3 and 0.10 (mu)mol (middot) g of dry matter content(sup-1) (middot) h(sup-1) under aerobic and anaerobic conditions, respectively, whereas xylose accumulation rates were only 10% of the glucose accumulation rates.     

The cellulosome concept as an efficient microbial strategy for the degradation of insoluble polysaccharides.

The cellulosome is an extracellular supramolecular machine that can efficiently degrade crystalline cellulosic substrates and associated plant cell wall polysaccharides. The cellulosome arrangement can also promote adhesion to the insoluble substrate, thus providing individual microbial cells with a direct competitive advantage in the utilization of the soluble hydrolysis products.     

Temperature Characteristics of Photosynthetic and Heterotrophic Activities: Seasonal Variations in Temperate Microbial Plankton

The seasonal variation in temperature characteristics of photosynthetic and heterotrophic activities in the microbial plankton of Bedford Basin, Nova Scotia, was investigated. Measurements were made of the photosynthetic uptake of [¹⁴C]bicarbonate and its incorporation into cellular protein as well as the heterotrophic uptake of ³H-labeled amino acids and their incorporation into cellular protein. Activity-temperature curves were analyzed objectively by nonlinear estimation of parameters from various mathematical models. Over the seasonal cycle, the cardinal temperatures and a parameter formally equivalent to the thermodynamic enthalpy of activation for most of the four processes measured were positively correlated with the water temperature. The temperature sensitivity of metabolic activity (i.e., change in activity per unit change in temperature) was indexed by the tangent to the activity-temperature curves. When this index was expressed in dimensionless form by normalization to the scaling factor of the activity-temperature curves, the resulting relative temperature sensitivity, evaluated at the prevailing temperature, proved to be statistically invariant throughout the year. During the height of the spring bloom, the water temperature (−0.3°C) was not so low as to inhibit metabolic activity of either the phytoplankton or the bacterioplankton. The evidence suggests that heterotrophic utilization of products is not suppressed during the spring phytoplankton bloom.     

海洋细菌CAZymes的研究进展

多糖是大型藻类、浮游植物和微生物的主要成分,多糖降解产物是海洋有机物的主要来源.碳水化合物活性酶(carbohydrate-active enzymes,CAZymes)是负责糖类化合物降解、修饰及生成糖苷键的功能酶系,是糖类物质代谢通路中的基本功能单元.多数异养细菌都具备一套完善的碳水化合物活性酶编码系统,它们是参与...     

Community development and modes of phosphorus utilization in a late summer ecosystem in the central Gulf of Finland, the Baltic Sea

The development of a filamentous, nitrogen-fixing cyanobacterial bloom was followed during July–August 1990 in a stratified basin in the central Gulf of Finland, Baltic Sea. Hydrography, dissolved inorganic, particulate and total nutrients, chlorophyll a, alkaline phosphatase activity, ³²PO₄-uptake and phytoplankton species were measured. The study period was characterized by wind-induced mixing events, followed by marked nutrient pulses and plankton community responses. Phosphate uptake was highest throughout the study period in the size fraction dominated by bacteria and picocyanobacteria (< 2 µm) and the proportion of uptake in the size fraction 2–10 µm remained low (2–6%). Higher phosphate turnover times were observed in a community showing signs of enhanced heterotrophic activity. The bloom of filamentous, nitrogen-fixing cyanobacteria Aphanizomenon flos-aquae was promoted by a nutrient pulse with an inorganic nutrient ratio (DIN:DIP) of 15. The results show that the quality, frequency and magnitude of the physically forced nutrient pulses have an important role in determining the relative share of the different modes of phosphorus utilization and hence in determining the cyanobacterial bloom intensity and species composition in the Baltic Sea.     

Differences in the substrate spectrum of extracellular enzymes in shallow lakes of differing trophic status

Organic matter fluxes and food web interactions in lakes depend on the abilities of heterotrophic microbial communities to access and degrade organic matter, a process that begins with extracellular hydrolysis of high molecular weight substrates. In order to determine whether patterns of enzymatic hydrolysis vary among shallow lakes of different trophic status, we investigated the hydrolysis of six specific organic macromolecules (polysaccharides) in the spring and late summer in four adjacent shallow lakes of eutrophic, oligotrophic, and dystrophic status in coastal North Carolina, USA. The spectrum of enzyme activities detected differed strongly between lakes, with all six polysaccharides hydrolyzed in West Mattamuskeet in May, while only two substrates were hydrolyzed in Lake Phelps in August/September. Differences in the spectrum of enzyme activities, and therefore the capabilities of heterotrophic microbial communities, were likely driven by variations among lakes in primary productivity patterns, sediment–water interactions, and/or water chemistry. Our data represent a first step towards a better understanding of carbon substrate availability and differences in carbon cycling pathways in shallow lakes of different trophic status.     

Sequential bioavailability of sedimentary organic matter to heterotrophic bacteria

Aquatic sediments harbour diverse microbial communities that mediate organic matter degradation and influence biogeochemical cycles. The pool of bioavailable carbon continuously changes as a result of abiotic processes and microbial activity. It remains unclear how microbial communities respond to heterogeneous organic matrices and how this ultimately affects heterotrophic respiration. To explore the relationships between the degradation of mixed carbon substrates and microbial activity, we incubated batches of organic‐rich sediments in a novel bioreactor (IsoCaRB) that permitted continuous observations of CO₂ production rates, as well as sequential sampling of isotopic signatures (δ¹³C, Δ¹⁴C), microbial community structure and diversity, and extracellular enzyme activity. Our results indicated that lower molecular weight (MW), labile, phytoplankton‐derived compounds were degraded first, followed by petroleum‐derived exogenous pollutants, and finally by higher MW polymeric plant material. This shift in utilization coincided with a community succession and increased extracellular enzyme activities. Thus, sequential utilization of different carbon pools induced changes at both the community and cellular level, shifting community composition, enzyme activity, respiration rates, and residual organic matter reactivity. Our results provide novel insight into the accessibility of sedimentary organic matter and demonstrate how bioavailability of natural organic substrates may affect the function and composition of heterotrophic bacterial populations.     

Seasonal microbial processes in a high-latitude fjord (Kongsfjorden, Svalbard): I. Heterotrophic bacteria, picoplankton and nanoflagellates

Temporal dynamics of the microbial food web in the Barents Sea and adjacent water masses in the European Arctic are to a large extent unknown. Seasonal variation in stocks and production rates of heterotrophic bacteria and phototrophic and heterotrophic picoplankton and nanoflagellates was investigated in the upper 50 m of the high-latitude Kongsfjorden, Svalbard, during six field campaigns between March and December 2006. Heterotrophic bacteria, picoplankton and nanoflagellates contributed to ecosystem structure and function in all seasons. Activity within the microbial food web peaked during spring bloom in April, parallel to low abundances of mesozooplankton. In the nutrient-limited post-bloom scenario, an efficient microbial loop, fuelled by dissolved organic carbon from abundant mesozooplankton feeding on phytoplankton and protozooplankton, facilitated maximum integrated primary production rates. A tight microbial food web consisting of heterotrophic bacteria and phototrophic and heterotrophic picoplankton and nanoflagellates was found in the stratified water masses encountered in July and September. Microbial stocks and rates were low but persistent under winter conditions. Seasonal comparisons of microbial biomass and production revealed that structure and function of the microbial food web were fundamentally different during the spring bloom when compared with other seasons. While the microbial food web was in a regenerative mode most of the time, during the spring bloom, a microbial transfer mode represented a trophic link for organic carbon in time and space. The microbial food web’s ability to fill different functional roles in periods dominated by new and regenerated production may enhance the ecological flexibility of pelagic ecosystems in the present era of climate change.     

Qualitative importance of the microbial loop and plankton community structure in a eutrophic lake during a bloom of cyanobacteria

Plankton community structure and major pools and fluxes of carbon were observed before and after culmination of a bloom of cyanobacteria in eutrophic Frederiksborg Slotssø, Denmark. Biomass changes of heterotrophic nanoflagellates, ciliates, microzooplankton (50 to 140 μm), and macrozooplankton (larger than 140 μm) were compared to phytoplankton and bacterial production as well as micro- and macrozooplankton ingestion rates of phytoplankton and bacteria. The carbon budget was used as a means to examine causal relationships in the plankton community. Phytoplankton biomass decreased and algae smaller than 20 μm replacedAphanizomenon after the culmination of cyanobacteria. Bacterial net production peaked shortly after the culmination of the bloom (510 μg C liter^?1 d^?1 and decreased thereafter to a level of approximately 124 μg C liter^?1 d^?1. Phytoplankton extracellular release of organic carbon accounted for only 4–9% of bacterial carbon demand. Cyclopoid copepods and small-sized cladocerans started to grow after the culmination, but food limitation probably controlled the biomass after the collapse of the bloom. Grazing of micro- and macrozooplankton were estimated from in situ experiments using labeled bacteria and algae. Macrozooplankton grazed 22% of bacterial net production during the bloom and 86% after the bloom, while microzooplankton (nauplii, rotifers and ciliates larger than 50 μm) ingested low amounts of bacteria and removed 10–16% of bacterial carbon. Both macro-and microzooplankton grazed algae smaller than 20 μm, although they did not control algal biomass. From calculated clearance rates it was found that heterotrophic nanoflagellates (40–440 ml^?1) grazed 3–4% of the bacterial production, while ciliates smaller than 50 μm removed 19–39% of bacterial production, supporting the idea that ciliates are an important link between bacteria and higher trophic levels. During and after the bloom ofAphanizomenon, major fluxes of carbon between bacteria, ciliates and crustaceans were observed, and heterotrophic nanoflagellates played a minor role in the pelagic food web.     

Impact of warming on phyto‐bacterioplankton coupling and bacterial community composition in experimental mesocosms

Global warming is assumed to alter the trophic interactions and carbon flow patterns of aquatic food webs. The impact of temperature on phyto‐bacterioplankton coupling and bacterial community composition (BCC) was the focus of the present study, in which an indoor mesocosm experiment with natural plankton communities from the western Baltic Sea was conducted. A 6°C increase in water temperature resulted, as predicted, in tighter coupling between the diatom‐dominated phytoplankton and heterotrophic bacteria, accompanied by a strong increase in carbon flow into bacterioplankton during the phytoplankton bloom phase. Suppressed bacterial development at cold in situ temperatures probably reflected lowered bacterial production and grazing by protists, as the latter were less affected by low temperatures. BCC was strongly influenced by the phytoplankton bloom stage and to a lesser extent by temperature. Under both temperature regimes, Gammaproteobacteria clearly dominated during the phytoplankton peak, with Glaciecola sp. as the single most abundant taxon. However, warming induced the appearance of additional bacterial taxa belonging to Betaproteobacteria and Bacteroidetes. Our results show that warming during an early phytoplankton bloom causes a shift towards a more heterotrophic system, with the appearance of new bacterial taxa suggesting a potential for utilization of a broader substrate spectrum.     

Two species of culturable bacteria associated with degradation of brown algae Fucus evanescens

The heterotrophic microbial enrichment community established during degradation of brown algae Fucus evanescens was characterized. A two-species bacterial community of marine culturable gamma-proteobacteria consisted of Pseudoalteromonas and Halomonas. The first member of the community, Pseudoalteromonas sp., was highly metabolically active, had bacteriolytic and hemolytic activities, produced proteinases (gelatinase and caseinase), lipases, DNases, and fucoidanhydrolases, laminaranases, alginases, pustulanases, beta-glucosidases, beta-galactosidases, beta-N-acetylglucosaminidases, and beta-xylosidases. The second member of the community, Halomonas marina, produced only caseinase and DNase, and it did not hydrolyze algal polysaccharides. Both members of the studied bacterial community utilized a range of easily assimilable monosaccharides and other low molecular weight organic substances. The results provide an evidence of the complex metabolic interrelations between two members of this culturable community. One of them Pseudoalteromonas sp., most likely plays the major role in the initial stages of algal degradation; the other one, H. marina, resistant to the bacteriolytic activity of the former, is able to utilize the products of degradation of polysaccharides.     

Relationship between succinate transport and production of extracellular poly(3-hydroxybutyrate) depolymerase in Pseudomonas lemoignei

The relationship between extracellular poly(3-hydroxybutyrate) (PHB) depolymerase synthesis and the unusual properties of a succinate uptake system was investigated in Pseudomonas lemoignei. Growth on and uptake of succinate were highly pH dependent, with optima at pH 5.6. Above pH 7, growth on and uptake of succinate were strongly reduced with concomitant derepression of PHB depolymerase synthesis. The specific succinate uptake rates were saturable by high concentrations of succinate, and maximal transport rates of 110 nmol/mg of cell protein per min were determined between pH 5.6 and 6. 8. The apparent KS0.5 values increased with increasing pH from 0.2 mM succinate at pH 5.6 to more than 10 mM succinate at pH 7.6. The uptake of [14C]succinate was strongly inhibited by several monocarboxylates. Dicarboxylates also inhibited the uptake of succinate but only at pH values near the dissociation constant of the second carboxylate function (pKa2). We conclude that the succinate carrier is specific for the monocarboxylate forms of various carboxylic acids and is not able to utilize the dicarboxylic forms. The inability to take up succinate2- accounts for the carbon starvation of P. lemoignei observed during growth on succinate at pH values above 7. As a consequence the bacteria produce high levels of extracellular PHB depolymerase activity in an effort to escape carbon starvation by utilization of PHB hydrolysis products.     

Uptake and turnover of acetate in hypersaline environments

Abstract: Acetate uptake and turnover rates were determined for the heterotrophic community in hypersaline environments (saltern crystallizer ponds, the Dead Sea) dominated by halpphilic Archaea. Acetate was formed from glycerol, which is potentially the major available carbon source for natural communities of halophilic Archaea. Values of [ K _t+ S _n] (the sum of the substrate affinity and the substrate concentration present in situ) for acetate measured in saltern crystallizer ponds were around 4.5–11.5 μM, while in the Dead Sea during a Dunaliella bloom values up to 12.8 μM were found. Maximal theoretical rates ( V _max) of acetate uptake in saltern crystallizer ponds were 12–56 nmol l⁻¹ h⁻¹, with estimated turnover times for acetate ( T _t) between 127–730 h at 35°C. V _max values measured in the Dead Sea were between 0.8 and 12.8 nmol l⁻¹ h⁻¹, with turnover times in the range of 320–2190 h. V _max values for acetate were much lower than those for glycerol. Comparisons with pure cultures of halophilic Archaea grown under different conditions showed that the natural communities were not adapted for preferential use of acetate. Both in natural brines and in pure cultures of halophilic Archaea, acetate incorporation rates rapidly decreased above the optimum pH value, probably since acetate enters the cell only in its unionized form. The low affinity for acetate, together with low potential utilization rates result in the long acetate turnover times, which explains the accumulation of acetate observed when low concentrations of glycerol are supplied as a nutrient to natural communities of halophilic Archaea.     

Vertical and seasonal variations of inorganic carbon allocation into macromolecules by phytoplankton population in a brown-colored and a clear-water lake

The aim of this study was to asses vertical and seasonal variations of inorganic carbon allocation into macromolecules by phytoplankton population in an humic and acidic lake (Lake Vassivière) and in a clearwater lake (Lake Pavin). Biochemical fractionation was done by consecutive differential extractions in order to separate proteins, polysaccharides, lipids and low molecular weight compounds (LMW) by virtue of their relative solubilities in different extraction solvents.Independent of depth and season, the principal photosynthetic end products were polysaccharides followed by proteins, LMW and lipids. However, inorganic carbon allocation into macromolecules varied, in these two lakes, with depth and with the taxonomic composition of phytoplankton. Carbon allocation into polysaccharides decreased with increasing depth, especially in the brown-colored humic lake, and Diatoms, showed high C incorporation into polysaccharides.     

Nutritional relationships among microorganisms in an epilithic biofilm community

Previous studies of an epilithic algal-bacterial community in a pristine mountain stream suggested that heterotrophic bacteria were responding to the metabolic activities of the phototrophic population. Subsequent studies were performed to follow the flow of labeled carbon, from its initial inorganic form, through the trophic levels of the mat community. A majority of primary production metabolites were excreted by the algal population during active growth; this shifted to an incorporation into cellular material as phototrophic activity declined. Results suggest that there was a direct flux of soluble algal products to the bacterial population, with little heterotrophic utilization of dissolved organics from the overlying stream water. Both phototrophic productivity and bacterial utilization of algal products peaked at approximately the same time of year. Activity of the diatom-dominated algal population declined as silica concentrations in the stream water dropped, leading to a situation in which the sessile bacteria were substrate limited. These events resulted in an almost complete disappearance of the community in early September.     

The submicron size fraction of seawater containing high numbers of virus particles as bioactive agent in unicellular plankton community successions

The effect of moderately (2.5 times) increasing the concentrationof the submicron size fraction (2–200 nm) of seawateron unicellular heterotrophic plankton organisms was tested ina series of experiments during algal bloom conditions. Usingelectron microscopy, vims-like particles (VLPs) were by farthe most visible component in this size fraction. We incubatedseawater. which was collected in late winter, in slowly rotatingcylindrical tanks that were exposed to spring temperature andlight conditions. This led to the onset of a mixed diatom bloomwhich was delayed by several days in submicron size-enrichedtreatments. Later, phytoplankton biomass reached values evenslightly higher than in unaltered incubations. The abundanceof heterotrophic unicellular plankton and VLPs exhibited morepronounced oscillation amplitudes during the early phase ofsuccession, thus being stimulated by additional submicron-sizedmaterial (including virions). VLPs are suggested as a possiblesupplementary diet for heterotrophic flagellates. From 190 honwards, toward the end of the incubation experiments (550 h).heterotrophic flagellate density was no longer influenced byinitial enrichment with the submicron size fraction. In contrast,bacteria were consistently repressed in the last two-thirdsof the experimental duration. Our results support the existenceof bioactive substances in the 2-200 nm size fraction, mostprobably virus particles, which are capable of influencing thedynamics and oscillation patterns in plankton community successionaccording to their concentration during algal bloom events.