Characterization of polyphosphate dynamics in the widespread freshwater diatom Achnanthidium minutissimum under varying phosphorus supplies.

Polyphosphates (polyP) are ubiquitous biomolecules that play a multitude of physiological roles in many cells. We have studied the presence and role of polyP in a unicellular alga, the freshwater diatom Achnanthidium minutissimum. This diatom stores up to 2.0 pg·cell-1 of polyP, with chain lengths ranging from 130 to 500 inorganic phosphate units (Pi). We applied energy dispersive X-ray spectroscopy, Raman/fluorescence microscopy, and biochemical assays to localize and characterize the intracellular polyP granules that were present in large apical vacuoles. We investigated the fate of polyP in axenic A. minutissimum cells grown under phosphorus (P), replete (P(+)), or P deplete (P(-)) cultivation conditions and observed that in the absence of exogenous P, A. minutissimum rapidly utilizes their internal polyP reserves, maintaining their intrinsic growth rates for up to 8 days. PolyP-depleted A. minutissimum cells rapidly took up exogenous P a few hours after Pi resupply and generated polyP three times faster than cells that were not initially subjected to P limitation. Accordingly, we propose that A. minutissimum deploys a succession of acclimation strategies regarding polyP dynamics where the production or consumption of polyP plays a central role in the homeostasis of the diatom.

Are drivers of root-associated fungal community structure context specific?

The composition and structure of plant-root-associated fungal communities are determined by local abiotic and biotic conditions. However, the relative influence and identity of relationships to abiotic and biotic factors may differ across environmental and ecological contexts, and fungal functional groups. Thus, understanding which aspects of root-associated fungal community ecology generalise across contexts is the first step towards a more predictive framework. We investigated how the relative importance of biotic and abiotic factors scale across environmental and ecological contexts using high-throughput sequencing (ca. 55 M Illumina metabarcoding sequences) of >260 plant-root-associated fungal communities from six UK salt marshes across two geographic regions (South-East and North-West England) in winter and summer. Levels of root-associated fungal diversity were comparable with forests and temperate grasslands, quadrupling previous estimates of salt-marsh fungal diversity. Whilst abiotic variables were generally most important, a range of site- and spatial scale-specific abiotic and biotic drivers of diversity and community composition were observed. Consequently, predictive models of diversity trained on one site, extrapolated poorly to others. Fungal taxa from the same functional groups responded similarly to the specific drivers of diversity and composition. Thus site, spatial scale and functional group are key factors that, if accounted for, may lead to a more predictive understanding of fungal community ecology.

Identifying metabolic pathways for production of extracellular polymeric substances by the diatom Fragilariopsis cylindrus inhabiting sea ice.

Diatoms are significant primary producers in sea ice, an ephemeral habitat with steep vertical gradients of temperature and salinity characterizing the ice matrix environment. To cope with the variable and challenging conditions, sea ice diatoms produce polysaccharide-rich extracellular polymeric substances (EPS) that play important roles in adhesion, cell protection, ligand binding and as organic carbon sources. Significant differences in EPS concentrations and chemical composition corresponding to temperature and salinity gradients were present in sea ice from the Weddell Sea and Eastern Antarctic regions of the Southern Ocean. To reconstruct the first metabolic pathway for EPS production in diatoms, we exposed Fragilariopsis cylindrus, a key bi-polar diatom species, to simulated sea ice formation. Transcriptome profiling under varying conditions of EPS production identified a significant number of genes and divergent alleles. Their complex differential expression patterns under simulated sea ice formation was aligned with physiological and biochemical properties of the cells, and with field measurements of sea ice EPS characteristics. Thus, the molecular complexity of the EPS pathway suggests metabolic plasticity in F. cylindrus is required to cope with the challenging conditions of the highly variable and extreme sea ice habitat.

Different Types of Diatom-Derived Extracellular Polymeric Substances Drive Changes in Heterotrophic Bacterial Communities from Intertidal Sediments.

Intertidal areas support extensive diatom-rich biofilms. Such microphytobenthic (MPB) diatoms exude large quantities of extracellular polymeric substances (EPS) comprising polysaccharides, glycoproteins and other biopolymers, which represent a substantial carbon pool. However, degradation rates of different EPS components, and how they shape heterotrophic communities in sediments, are not well understood. An aerobic mudflat-sediment slurry experiment was performed in the dark with two different EPS carbon sources from a diatom-dominated biofilm: colloidal EPS (cEPS) and the more complex hot-bicarbonate-extracted EPS. Degradation rate constants determined over 9 days for three sediment fractions [dissolved organic carbon (DOC), total carbohydrates (TCHO), and (cEPS)] were generally higher in the colloidal-EPS slurries (0.105-0.123 d-1) compared with the hot-bicarbonate-extracted-EPS slurries (0.060-0.096 d-1). Addition of hot-bicarbonate-EPS resulted in large increases in dissolved nitrogen and phosphorous by the end of the experiment, indicating that the more complex EPS is an important source of regenerated inorganic nutrients. Microbial biomass increased ~4-6-fold over 9 days, and pyrosequencing of bacterial 16S rRNA genes revealed that the addition of both types of EPS greatly altered the bacterial community composition (from 0 to 9 days) compared to a control with no added EPS. Bacteroidetes (especially Tenacibaculum) and Verrucomicrobia increased significantly in relative abundance in both the hot-bicarbonate-EPS and colloidal-EPS treatments. These differential effects of EPS fractions on carbon-loss rates, nutrient regeneration and microbial community assembly improve our understanding of coastal-sediment carbon cycling and demonstrate the importance of diverse microbiota in processing this abundant pool of organic carbon.

Soil compartment is a major determinant of the impact of simulated rainfall on desert microbiota.

Although desert soils support functionally important microbial communities that affect plant growth and influence many biogeochemical processes, the impact of future changes in precipitation patterns on the microbiota and their activities is largely unknown. We performed in-situ experiments to investigate the effect of simulated rainfall on bacterial communities associated with the widespread perennial shrub, Rhazya stricta in Arabian desert soils. The bacterial community composition was distinct between three different soil compartments: surface biological crust, root-attached, and the broader rhizosphere. Simulated rainfall had no significant effect on the overall bacterial community composition, but some population-level responses were observed, especially in soil crusts where Betaproteobacteria, Sphingobacteria, and Bacilli became more abundant. Bacterial biomass in the nutrient-rich crust increased three-fold one week after watering, whereas it did not change in the rhizosphere, despite its much higher water retention. These findings indicate that between rainfall events, desert-soil microbial communities enter into stasis, with limited species turnover, and reactivate rapidly and relatively uniformly when water becomes available. However, microbiota in the crust, which was relatively enriched in nutrients and organic matter, were primarily water-limited, compared with the rhizosphere microbiota that were co-limited by nutrients and water.

Protection of cells from salinity stress by extracellular polymeric substances in diatom biofilms.

Diatom biofilms are abundant in the marine environment. It is assumed (but untested) that extracellular polymeric substances (EPS), produced by diatoms, enable cells to cope with fluctuating salinity. To determine the protective role of EPS, Cylindrotheca closterium was grown in xanthan gum at salinities of 35, 50, 70 and 90 ppt. A xanthan matrix significantly increased cell viability (determined by SYTOX-Green), growth rate and population density by up to 300, 2,300 and 200%, respectively. Diatoms grown in 0.75% w/v xanthan, subjected to acute salinity shock treatments (at salinities 17.5, 50, 70 and 90 ppt) maintained photosynthetic capacity, Fq'/Fm', within 4% of pre-shock values, whereas Fq'/Fm' in cells grown without xanthan declined by up to 64% with hypersaline shock. Biofilms that developed in xanthan at standard salinity helped cells to maintain function during salinity shock. These results provide evidence of the benefits of living in an EPS matrix for biofilm diatoms.

Broad-scale predictability of carbohydrates and exopolymers in Antarctic and Arctic sea ice.

Sea ice can contain high concentrations of dissolved organic carbon (DOC), much of which is carbohydrate-rich extracellular polymeric substances (EPS) produced by microalgae and bacteria inhabiting the ice. Here we report the concentrations of dissolved carbohydrates (dCHO) and dissolved EPS (dEPS) in relation to algal standing stock [estimated by chlorophyll (Chl) a concentrations] in sea ice from six locations in the Southern and Arctic Oceans. Concentrations varied substantially within and between sampling sites, reflecting local ice conditions and biological content. However, combining all data revealed robust statistical relationships between dCHO concentrations and the concentrations of different dEPS fractions, Chl a, and DOC. These relationships were true for whole ice cores, bottom ice (biomass rich) sections, and colder surface ice. The distribution of dEPS was strongly correlated to algal biomass, with the highest concentrations of both dEPS and non-EPS carbohydrates in the bottom horizons of the ice. Complex EPS was more prevalent in colder surface sea ice horizons. Predictive models (validated against independent data) were derived to enable the estimation of dCHO concentrations from data on ice thickness, salinity, and vertical position in core. When Chl a data were included a higher level of prediction was obtained. The consistent patterns reflected in these relationships provide a strong basis for including estimates of regional and seasonal carbohydrate and dEPS carbon budgets in coupled physical-biogeochemical models, across different types of sea ice from both polar regions.

Differences between aerobic and anaerobic degradation of microphytobenthic biofilm-derived organic matter within intertidal sediments.

Within intertidal sediments, much of the dissolved organic carbon (DOC) consists of carbohydrate-rich extracellular polymeric substances (EPS) produced by microphytobenthic biofilms. EPS are an important source of carbon and energy for aerobic and anaerobic microorganisms owing to burial of microphytobenthos and downward transport of their exudates. We established slurries of estuarine biofilms to determine the fate of organic carbon and EPS fractions, differing in size and complexity, under oxic and anoxic conditions. DOC and hot-water-extracted organic matter (predominately diatom chrysolaminarin) were utilised rapidly at similar rates in both conditions. Concentrations of insoluble, high-molecular-weight EPS were unchanged in oxic microcosms, but were significantly degraded under anoxic conditions (39% degradation by day 25). Methanogenesis and sulphate reduction were major anaerobic processes in the anoxic slurries, and 16S rRNA gene pyrosequencing revealed that Desulfobacteraceae (relative sequence abundance increased from 1.9% to 12.2%) and Desulfobulbaceae (increased from 1.5% to 4.3%) were the main sulphate reducers, whilst Clostridia and Bacteroidetes were likely responsible for anaerobic hydrolysis and fermentation of EPS. We conclude that a diverse consortium of anaerobic microorganisms (including coexisting sulphate reducers and methanogens) degrade both labile and refractory microphytobenthic-derived carbon and that anaerobic degradation may be the primary fate of more structurally complex components of microphytobenthic EPS.

Impact of a simulated oil spill on benthic phototrophs and nitrogen-fixing bacteria in mudflat mesocosms.

Coastal and estuarine ecosystems are highly susceptible to crude oil pollution. Therefore, in order to examine the resilience of benthic phototrophs that are pivotal to coastal ecosystem functioning, we simulated an oil spill in tidal mesocosms consisting of intact sediment cores from a mudflat at the mouth of the Colne Estuary, UK. At day 21, fluorescence imaging revealed a bloom of cyanobacteria on the surface of oiled sediment cores, and the upper 1.5 cm thick sediment had 7.2 times more cyanobacterial and 1.7 times more diatom rRNA sequences when treated with oil. Photosystem II operating efficiency (Fq'/Fm') was significantly reduced in oiled sediments at day 7, implying that the initial diatom-dominated community was negatively affected by oil, but this was no longer apparent by day 21. Oil addition significantly reduced numbers of the key deposit feeders, and the decreased grazing pressure is likely to be a major factor in the increased abundance of both diatoms and cyanobacteria. By day 5 concentrations of dissolved inorganic nitrogen were significantly lower in oiled mesocosms, likely resulting in the observed increase in nifH-containing, and therefore potentially dinitrogen-fixing, cyanobacteria. Thus, indirect effects of oil, rather than direct inhibition, are primarily responsible for altering the microphytobenthos.

Central role of dynamic tidal biofilms dominated by aerobic hydrocarbonoclastic bacteria and diatoms in the biodegradation of hydrocarbons in coastal mudflats.

Mudflats and salt marshes are habitats at the interface of aquatic and terrestrial systems that provide valuable services to ecosystems. Therefore, it is important to determine how catastrophic incidents, such as oil spills, influence the microbial communities in sediment that are pivotal to the function of the ecosystem and to identify the oil-degrading microbes that mitigate damage to the ecosystem. In this study, an oil spill was simulated by use of a tidal chamber containing intact diatom-dominated sediment cores from a temperate mudflat. Changes in the composition of bacteria and diatoms from both the sediment and tidal biofilms that had detached from the sediment surface were monitored as a function of hydrocarbon removal. The hydrocarbon concentration in the upper 1.5 cm of sediments decreased by 78% over 21 days, with at least 60% being attributed to biodegradation. Most phylotypes were minimally perturbed by the addition of oil, but at day 21, there was a 10-fold increase in the amount of cyanobacteria in the oiled sediment. Throughout the experiment, phylotypes associated with the aerobic degradation of hydrocarbons, including polycyclic aromatic hydrocarbons (PAHs) (Cycloclasticus) and alkanes (Alcanivorax, Oleibacter, and Oceanospirillales strain ME113), substantively increased in oiled mesocosms, collectively representing 2% of the pyrosequences in the oiled sediments at day 21. Tidal biofilms from oiled cores at day 22, however, consisted mostly of phylotypes related to Alcanivorax borkumensis (49% of clones), Oceanospirillales strain ME113 (11% of clones), and diatoms (14% of clones). Thus, aerobic hydrocarbon biodegradation is most likely to be the main mechanism of attenuation of crude oil in the early weeks of an oil spill, with tidal biofilms representing zones of high hydrocarbon-degrading activity.

High site fidelity and low site connectivity in temperate salt marsh fish populations: a stable isotope approach.

Adult and juvenile fish utilise salt marshes for food and shelter at high tide, moving into adjacent sublittoral regions during low tide. Understanding whether there are high levels of site fidelity for different species of coastal fish has important implications for habitat conservation and the design of marine protected areas. We hypothesised that common salt marsh fish species would demonstrate a high site fidelity, resulting in minimal inter-marsh connectivity. Carbon ((13)C) and nitrogen ((15)N) stable isotope ratios of larvae and juveniles of five common salt marsh fish (Atherina presbyter, Chelon labrosus, Clupea harengus, Dicentrarchus labrax, Pomatoschistus microps), seven types of primary producer and seven secondary consumer food sources were sampled in five salt marshes within two estuary complexes along the coast of south-east England. Significant differences in (13)C and (15)N signatures between salt marshes indicated distinct sub-populations utilising the area of estuary around each salt marsh, and limited connectivity, even within the same estuary complex. (15)N ratios were responsible for the majority of inter-marsh differences for each species and showed similar site-specific patterns in ratios in primary producers, secondary consumers and fish. Fish diets (derived from isotope mixing models) varied between species but were mostly consistent between marsh sites, indicating that dietary shifts were not the source of variability of the inter-marsh isotopic signatures within species. These results demonstrate that for some common coastal fish species, high levels of site fidelity result in individual salt marshes operating as discrete habitats for fish assemblages.

Production and Characterization of the Intra- and Extracellular Carbohydrates and Polymeric Substances (EPS) of Three Sea-Ice Diatom Species, and Evidence for a Cryoprotective Role for EPS.

Diatoms and their associated extracellular polymeric substances (EPS) are major constituents of the microalgal assemblages present within sea ice. Yields and chemical composition of soluble and cell-associated polysaccharides produced by three sea-ice diatoms, Synedropsis sp., Fragilariopsis curta, and F. cylindrus, were compared. Colloidal carbohydrates (CC) contained heteropolysaccharides rich in mannose, xylose, galactose, and glucose. Synedropsis sp. CC consisted mainly of carbohydrates <8 kDa size, with relatively soluble EPS, compared to high proportions of less-soluble EPS produced by both Fragilariopsis spp. F. curta colloidal EPS contained high concentrations of amino sugars (AS). Both Fragilariopsis species had high yields of hot bicarbonate (HB) soluble EPS, rich in xylose, mannose, galactose, and fucose (and AS in F. cylindrus). All species had frustule-associated EPS rich in glucose-mannose. Nutrient limitation resulted in declines in EPS yields and in glucose content of all EPS fractions. Significant similarities between EPS fractions from cultures and different components of natural EPS from Antarctic sea ice were found. Increased salinity (52) reduced growth, but increased yields of EPS in Fragilariopsis cylindrus. Ice formation was inhibited byF. cylindrus, EPS, and by enhanced EPS content (additional xanthan gum) down to -12°C, with growth rate reduced in the presence of xanthan. Differences in the production and composition of EPS between Synedropsis sp. and Fragilariopsis spp., and the association between EPS, freezing and cell survival, supports the hypothesis that EPS production is a strategy to assist polar ice diatoms to survive the cold and saline conditions present in sea ice.

CALCIUM RELEASE FROM INTRACELLULAR STORES IS NECESSARY FOR THE PHOTOPHOBIC RESPONSE IN THE BENTHIC DIATOM NAVICULA PERMINUTA (BACILLARIOPHYCEAE)(1).

Complex photoreceptor pathways exist in algae to exploit light as a sensory stimulus. Previous studies have implicated calcium in blue-light signaling in plants and algae. A photophobic response to high-intensity blue light was characterized in the marine benthic diatom Navicula perminuta (Grunow) in van Heurck. Calcium modulators were used to determine the involvement of calcium in the signaling of this response, and the fluorescent calcium indicator Calcium Crimson was used to image changes in intracellular [Ca(2+) ] during a response. A localized, transient elevation of Calcium Crimson fluorescence was seen at the cell tip at the time of cell reversal. Intracellular calcium release inhibitors produced a significant decrease in the population photophobic response. Treatments known to decrease influx of extracellular calcium had no effect on the population photophobic response but did cause a significant decrease in average cell speed. As the increase in intracellular [Ca(2+) ] at the cell tip corresponded to the time of direction change rather than the onset of the light stimulus, it would appear that Ca(2+) constitutes a component of the switching mechanism that leads to reversal of the locomotion machinery. Our current evidence suggests that the source of this Ca(2+) is intracellular.

Resistance and resilience of benthic biofilm communities from a temperate saltmarsh to desiccation and rewetting.

Periods of desiccation and rewetting are regular, yet stressful events encountered by saltmarsh microbial communities. To examine the resistance and resilience of microbial biofilms to such stresses, sediments from saltmarsh creeks were allowed to desiccate for 23 days, followed by rewetting for 4 days, whereas control sediments were maintained under a natural tidal cycle. In the top 2 mm of the dry sediments, salinity increased steadily from 36 to 231 over 23 days, and returned to seawater salinity on rewetting. After 3 days, desiccated sediments had a lower chlorophyll a (Chl a) fluorescence signal as benthic diatoms ceased to migrate to the surface, with a recovery in cell migration and Chl a fluorescence on rewetting. Extracellular ß-glucosidase and aminopeptidase activities decreased within the first week of drying, but increased sharply on rewetting. The bacterial community in the desiccating sediment changed significantly from the controls after 14 days of desiccation (salinity 144). Rewetting did not cause a return to the original community composition, but led to a further change. Pyrosequencing analysis of 16S rRNA genes amplified from the sediment revealed diverse microbial responses, for example desiccation enabled haloversatile Marinobacter species to increase their relative abundance, and thus take advantage of rewetting to grow rapidly and dominate the community. A temporal sequence of effects of desiccation and rewetting were thus observed, but the most notable feature was the overall resistance and resilience of the microbial community.

Solutes determine the temperature windows for microbial survival and growth.

Microbial cells, and ultimately the Earth's biosphere, function within a narrow range of physicochemical conditions. For the majority of ecosystems, productivity is cold-limited, and it is microbes that represent the failure point. This study was carried out to determine if naturally occurring solutes can extend the temperature windows for activity of microorganisms. We found that substances known to disorder cellular macromolecules (chaotropes) did expand microbial growth windows, fungi preferentially accumulated chaotropic metabolites at low temperature, and chemical activities of solutes determined microbial survival at extremes of temperature as well as pressure. This information can enhance the precision of models used to predict if extraterrestrial and other hostile environments are able to support life; furthermore, chaotropes may be used to extend the growth windows for key microbes, such as saprotrophs, in cold ecosystems and man-made biomes.

Microphytobenthos and phytoplankton in the Severn estuary, UK: present situation and possible consequences of a tidal energy barrage.

Information on the distribution of microphytobenthos (micro-algae forming biofilms on sediment surfaces) and phytoplankton in the Severn estuary is reviewed. Microphytobenthos (MPB) are widely distributed in salt marsh and mudflat environments, with biomass levels lower than in other estuaries (average 53 mg chl am(-2) on mud, 12 mg chl am(-2) on sand). Seasonal and spatial patterns occur in the species composition of biofilms. Large areas of the Severn have not been surveyed, but it is likely that MPB are abundant in these regions. Dissolved inorganic N, P and Si concentrations are high in the upper estuary (>400 microM nitrate, >10 microM phosphate, >140 microM silicate) and decrease seaward. Phytoplanktonic chl a concentrations are low in the main estuary (2.2 microg chl aL(-1)), but increase in the Bristol Channel and at the head of the estuary (>10 microg chl aL(-1)). High turbidity is the likely cause for low phytoplankton activity. Annual production of MPB was estimated at 33 g Cm(-2) of inter-tidal area y(-1) in the Severn estuary. This estimated benthic primary production would utilise 3.9% and 4.9% of the annual estuarine N and P loads. Construction of tidal energy barrages is estimated to significantly reduce annual estuarine MPB production (by 77% for the Cardiff-Weston barrage). It is unlikely that any potential increases in MPB biomass on remaining inter-tidal areas will be sufficient to compensate for these losses. In general, the data coverage for benthic and planktonic distribution, production and related nutrient cycles is extremely limited and significant new research work is needed to enable more definite predictions of the post-barrage situation to be made.

LIGHT-INDUCED MOTILE RESPONSES OF THE ESTUARINE BENTHIC DIATOMS NAVICULA PERMINUTA AND CYLINDROTHECA CLOSTERIUM (BACILLARIOPHYCEAE)(1).

Motility of estuarine epipelic (mud-inhabiting) diatoms is an important adaptation to living in biofilms present within fine sediments. Motility allows cells to migrate within the photic zone in response to a wide range of environmental stimuli. The motile responses of two species of benthic diatoms to photon fluence rates and spectral quality were investigated. Cultures of Navicula perminuta (Grunow) in van Heurck and Cylindrotheca closterium (Ehrenb.) J. C. Lewin et Reimann both exhibited photoaccumulation at ∼200 µmol · m(-2) · s(-1) and photodispersal from photon flux densities (PFDs) of ∼15 µmol · m(-2) · s(-1) . Photokinesis (changing cell speed) contributed toward photodispersal for both species, and red light (λ = 681-691 nm) was most effective at inducing this process. N. perminuta showed a phototactic (directional) response, with active movement in response to a light gradient. Although this response was exhibited in white light, these directional responses were only elicited by wavelengths from 430 to 510 nm. In contrast, C. closterium did not exhibit phototaxis under any light conditions used in this study. Motile benthic diatoms thus exhibit complex and sophisticated responses to light quantity and quality, involving combinations of photokinesis and phototaxis, which can contribute toward explaining the patterns of large-scale cell movements observed in natural estuarine biofilms.

Diatom-derived carbohydrates as factors affecting bacterial community composition in estuarine sediments.

Microphytobenthic biofilms in estuaries, dominated by epipelic diatoms, are sites of high primary productivity. These diatoms exude large quantities of extracellular polymeric substances (EPS) comprising polysaccharides and glycoproteins, providing a substantial pool of organic carbon available to heterotrophs within the sediment. In this study, sediment slurry microcosms were enriched with either colloidal carbohydrates or colloidal EPS (cEPS) or left unamended. Over 10 days, the fate of these carbohydrates and changes in beta-glucosidase activity were monitored. Terminal restriction fragment length polymorphism (T-RFLP), DNA sequencing, and quantitative PCR (Q-PCR) analysis of 16S rRNA sequences were used to determine whether sediment bacterial communities exhibited compositional shifts in response to the different available carbon sources. Initial heterotrophic activity led to reductions in carbohydrate concentrations in all three microcosms from day 0 to day 2, with some increases in beta-glucosidase activity. During this period, treatment-specific shifts in bacterial community composition were not observed. However, by days 4 and 10, the bacterial community in the cEPS-enriched sediment diverged from those in colloid-enriched and unamended sediments, with Q-PCR analysis showing elevated bacterial numbers in the cEPS-enriched sediment at day 4. Community shifts were attributed to changes in cEPS concentrations and increased beta-glucosidase activity. T-RFLP and sequencing analyses suggested that this shift was not due to a total community response but rather to large increases in the relative abundance of members of the gamma-proteobacteria, particularly Acinetobacter-related bacteria. These experiments suggest that taxon- and substrate-specific responses within the bacterial community are involved in the degradation of diatom-derived extracellular carbohydrates.

Impact of elevated UV-B radiation on photosynthetic electron transport, primary productivity and carbon allocation in estuarine epipelic diatoms.

Epipelic diatoms are important components of microphytobenthic biofilms. Cultures of four diatom species (Amphora coffeaeformis, Cylindrotheca closterium, Navicula perminuta and Nitzschia epithemioides) and assemblages of mixed diatom species collected from an estuary were exposed to elevated levels of ultraviolet-B (UV-B) radiation. Short exposures to UV-B resulted in decreases in photosystem II (PSII) photochemistry, photosynthetic electron transport, photosynthetic carbon assimilation and changes in the pattern of allocation of assimilated carbon into soluble colloidal, extracellular polysaccharides (EPS) and glucan pools. The magnitude of the effects of the UV-B treatments varied between species and was also dependent upon the photosynthetically active photon flux density (PPFD) to which the cells were also exposed, with effects being greater at lower light levels. Both increases in nonphotochemical quenching of excitation energy in the pigment antennae and photodamage to the D1 reaction centres contributed to decreases in PSII photochemistry. All species demonstrated a rapid ability to recover from perturbations of PSII photochemistry, with some species recovering during the UV-B exposure period. Some of the perturbations induced in carbon metabolism were independent of effects on PSII photochemistry and photosynthetic electron transport. Elevated UV-B can significantly inhibit photosynthetic performance, and modify carbon metabolism in epipelic diatoms. However, the ecological effects of UV-B at the community level are difficult to predict as large variations occur between species.

Nitrous oxide formation in the Colne estuary, England: the central role of nitrite.

Nitrate and nitrite concentrations in the water and nitrous oxide and nitrite fluxes across the sediment-water interface were measured monthly in the River Colne estuary, England, from December 1996 to March 1998. Water column concentrations of N(2)O in the Colne were supersaturated with respect to air, indicating that the estuary was a source of N(2)O for the atmosphere. At the freshwater end of the estuary, nitrous oxide effluxes from the sediment were closely correlated with the nitrite concentrations in the overlying water and with the nitrite influx into the sediment. Increases in N(2)O production from sediments were about 10 times greater with the addition of nitrite than with the addition of nitrate. Rates of denitrification were stimulated to a larger extent by enhanced nitrite than by nitrate concentrations. At 550 microM nitrite or nitrate (the highest concentration used), the rates of denitrification were 600 micromol N.m(-2).h(-1) with nitrite but only 180 micromol N.m(-2).h(-1) with nitrate. The ratios of rates of nitrous oxide production and denitrification (N(2)O/N(2) x 100) were significantly higher with the addition of nitrite (7 to 13% of denitrification) than with nitrate (2 to 4% of denitrification). The results suggested that in addition to anaerobic bacteria, which possess the complete denitrification pathway for N(2) formation in the estuarine sediments, there may be two other groups of bacteria: nitrite denitrifiers, which reduce nitrite to N(2) via N(2)O, and obligate nitrite-denitrifying bacteria, which reduce nitrite to N(2)O as the end product. Consideration of free-energy changes during N(2)O formation led to the conclusion that N(2)O formation using nitrite as the electron acceptor is favored in the Colne estuary and may be a critical factor regulating the formation of N(2)O in high-nutrient-load estuaries.