THE COMPLEX EXTRACELLULAR POLYSACCHARIDES OF MAINLY CHAIN‐FORMING FRESHWATER DIATOM SPECIES FROM EPILITHIC BIOFILMS1

Diatoms are dominant organisms in phototrophic biofilms in aquatic habitats. They produce copious amounts of extracellular polymeric substances (EPS), which mainly consist of carbohydrates and traces of proteins and glycoproteins. This study focuses on the characterization of EPS from a total of 14 diatoms belonging to the six genera Achnanthes, Cymbella, Fragilaria, Punctastriata, Staurosira, and Pseudostaurosira, all of which were isolated from epilithic biofilms of the littoral zone of Lake Constance. EPS from all isolates were extracted by a sequential extraction procedure resulting in five different fractions. The monosaccharide composition of each fraction was analyzed by HPLC equipped with a pulse amperiometric detector, yielding results similar to those obtained by probing the EPS structures with monomer‐specific fluorophore‐linked lectins. Significant differences in carbohydrate composition occurred in the different fractions of single isolates. Most of the diatom isolates in our study form chain‐like colonies in which the cells are attached to each other by intercellular pads. Here we demonstrate that these pads can be dissolved in hot bicarbonate and that they show a heterogeneous composition of monosaccharides in contrast to other fractions, which mostly were dominated by one or two monosaccharides. Principal component analysis indicates a correlation between carbohydrate composition of EPS fractions and the phylogenetic relationship of the respective species, indicating that EPS analyses under defined culture conditions may support taxonomic analyses.     

Carbohydrate Production in Relation to Microphytobenthic Biofilm Development: An Integrated Approach in a Tidal Mesocosm

Experiments were performed to evaluate short-term changes in sediment extracellular carbohydrates for a multispecific assemblage of benthic diatoms in relation to physiological status, endogenous migratory rhythms, and environmental conditions. For this purpose, a mesocosm was used, which simulated both tidal and dark: light alternating cycles under controlled conditions. Scanning electronic microscopy in combination with picture analyses indicated that natural diatom migration patterns were reproduced in the mesocosm. Two EPS fractions were operationally separated in colloidal carbohydrate measurements: alcohol-soluble EPS (termed “soluble EPS”) and alcohol-insoluble EPS (termed “bound EPS”). Microphytobenthic biomass followed a logistic-type curve and converged toward a maximal value termed the “biotic capacity of the local environment.” Both EPS fractions showed oscillations with production during photosynthetic periods and sharp decreases during night immersion periods. Productions of both EPS fractions increased with Chl a production during light periods suggesting a light dependence in relation to migratory patterns. The decreases in both EPS fractions, which occurred during night immersion periods suggest that carbohydrate hydrolysis and/or washaway affected both EPS fractions similarly in benthic environments. Our results confirm the theory according to which the two distinct fractions are under different metabolic controls. No change in soluble EPS release was obtained during the transition from logarithmic to stationary phase. On the other hand, a metabolism modification of microalgae, probably related to ammonium depletion, occurred when cells entered the stationary phase, since there was a high enhancement in bound EPS production. Mesocosm results can serve as a system of reference useful to characterize biofilm development in field investigations and to revisit the effective implication of each EPS fraction in sediment stability.     

Studies on the phytoplankton ecology of the Trondheimsfjord. I. The chemical composition of phytoplankton populations

Samples of phytoplankton populations from the Trondheimsfjord, collected in 1970 and the first five months of 1971, have been analysed for carbohydrate, protein, lipid, and phosphorus. Lipid was in all cases less than 10% of the organic dry matter. The $\text{N}\text{P}$ ratio was remarkably constant, but the ratio protein/carbohydrate varied between wide limits. For samples consisting mainly of dinoflagellates, the protein/carbohydrate ratio was always low, due to a large amount of insoluble polysaccharides, probably corresponding to material in the cell walls.For diatoms, the carbohydrates may conveniently be divided into three fractions: 1) an acidsoluble glucan of the β-1, 3-linked type; 2) an alkali-soluble fraction giving a complex mixture of monosaccharides on hydrolysis and, 3) an insoluble glucan. The amounts of acid-soluble glucan varied from 7.7 to 36.5% of organic dry matter and these changes are the main cause of the variation of the protein/carbohydrate ratio of diatom samples. For diatom samples this ratio is a valuable indicator of the physiological state of the population. The variations observed in this study are discussed.     

Molecular characterization of total organic matter and carbohydrates in peat samples from a cypress swamp by pyrolysis-mass spectrometry and wet-chemical methods

The organic matter present in coarse- and fine-grained sample preparations of a peat core from Minnie's Lake, Okefenokee Swamp (Georgia, USA) was analysed on a molecular level by gas chromatography-mass spectrometry of alditol acetates obtained from polysaccharides after acid hydrolysis and derivatization and by flash pyrolysis-mass spectrometry. We found that the coarse-grained fraction is dominated by neutral monosaccharides derived from vascular plants whilst the fine-grained fraction is dominated by neutral monosaccharides derived from microorganisms. Relatively high contributions of xylose discriminate the Taxodium derived peat from the underlying peats derived from Carex and Nymphaea which show relatively low contributions of this sugar. Factor-discriminant analysis of the Py-MS data indicates that most carbohydrate degradation is restricted to the upper peat levels and that guaiacyl and syringyl type lignins are better preserved than carbohydrates. Resinous organic matter - presumably derived from Taxodium — is selectively preserved in the fine-grained fraction. Deviant monosaccharide concentrations and Py-MS results observed for the top sample of the Carex peat were explained by oxygenation of the peat, a concomitantly high microbial activity and subsequent preservation of the organic material due to the action of phytotoxins derived from Taxodium.     

THE COMPLEX POLYSACCHARIDES OF THE RAPHID DIATOM PINNULARIA VIRIDIS (BACILLARIOPHYCEAE)1

A combination of carbohydrate analysis and atomic force microscopy (AFM) was used to characterize the polysaccharides of the pennate diatom, Pinnularia viridis (Nitzsch) Ehrenberg. Polymeric substances were fractionated into those in the spent culture medium (SCM) and those sequentially extracted from the cells with water at 45° C (WW), NaHCO₃ containing EDTA at 95° C (HB), and 1 M NaOH containing NaBH₄ at 95° C. Carbohydrate, protein, and sulfate were detected in all the fractions, but their relative proportions differed significantly. Nineteen sugars were identified, including pentoses, hexoses, 6‐deoxyhexoses, O‐methylated sugars, aminohexoses, and traces of uronic acids. To some extent, the same constituent monosaccharides and a proportion of the linkage patterns occurred in all four fractions, indicating the fractions contained a spectrum of highly heterogeneous but structurally related polysaccharides. Several carbohydrates were enriched in specific fractions. A soluble, partially substituted, 3‐linked galactan was slightly enriched in the SCM. The WW fraction was highly enriched in 3‐linked glucan, presumably derived from chrysolaminaran. Chemical and AFM data for the WW and HB fractions indicated that compositional differences were associated with substantial changes in the morphology and properties of the cell surface mucilage. Soluble polymers relatively enriched in fucose conferred a degree of softness and compressibility to the mucilage, whereas most of the mucilage comprised firmer more gelatinous polymers comparatively enriched in rhamnose. The frustule residue dissolved during extraction with NaOH, and a partially substituted 3‐linked mannan, together with relatively large amounts of protein, was obtained.     

Intestinal inflammation alters mucosal carbohydrate foraging and monosaccharide incorporation into microbial glycans

Endogenous carbohydrates released from the intestinal mucus represent a constant source of nutrients to the intestinal microbiota. Mucus‐derived carbohydrates can also be used as building blocks in the biosynthesis of bacterial cell wall components, thereby influencing host mucosal immunity. To assess the uptake of endogenous carbohydrates by gut microbes in healthy mice and during intestinal inflammation, we applied azido‐monosaccharides that can be tracked on bacterial cell walls after conjugation with fluorophores. In interleukin‐10 deficient mice, changes in the gut microbiota were accompanied by decreased carbohydrate hydrolase activities and increased lumenal concentrations of host glycan‐derived monosaccharides. Tracking of the monosaccharide N‐azidoacetylglucosamine (GlcNAz) in caecum bacteria revealed a preferential incorporation of this carbohydrate by Xanthomonadaceae in healthy mice and by Bacteroidaceae in interleukin‐10 deficient mice. These GlcNAz‐positive Bacteroidaceae fractions mainly belonged to the species B. acidifaciens and B. vulgatus. Growth of Bacteroides species in the presence of specific monosaccharides changed their stimulatory activity toward CD11c⁺ dendritic cells. Expression of activation markers and cytokine production was highest after stimulation of dendritic cells with B. vulgatus. The variable incorporation of monosaccharides by related Bacteroides species underline the necessity to investigate intestinal bacteria down to the species level when addressing microbiota‐host interactions.     

Carbohydrate synthesis by two Navicula strains isolated from benthic and pelagic mucilages in the Tyrrhenian Sea (Tuscan Archipelago)

Two exopolysaccharide (EPS)-producing strains of the diatom Navicula, were isolated from benthic and pelagic mucilaginous aggregates sampled in the Tyrrhenian Sea and cultured under laboratory conditions. The amount of carbohydrate formed over the growth period and on a per cell basis was quite similar. However, the benthic strain showed a preferential synthesis of the bound (i.e., cellular and capsular) carbohydrate fraction, whereas the pelagic strain preferentially synthesised soluble, polymeric carbohydrates. The polysaccharides released into the medium by the two strains showed the same qualitative monosaccharidic composition, being constituted by two acidic and six neutral sugars. It is suggested that the difference between the benthic and the pelagic strain in the synthesis of bound or soluble carbohydrates may be related to the different role of these compounds in the particular habitats of the strains.     

Colloidal organic carbon and trace metal (Cd, Fe, and Zn) releases by diatom exudation and copepod grazing

Colloidal macromolecular organic compounds are important intermediaries between solution and particle phases and play a critical role in the biogeochemistry of trace metals and organic carbon. The releases of colloidal organic carbon and trace metals (Cd, Fe, and Zn) mediated by copepod grazing and decomposition, and direct diatom exudation, were examined using a radiotracer approach. The colloidal phase was operationally defined in this study as the size fraction between 5 kDa and 0.2 μm and the dissolved phase as the ≤0.2 μm filter passing phase. About 13-60% of dissolved carbon exuded by the diatom Thalassiosira pseudonana was partitioned into the colloidal phase, and this fraction increased considerably as the diatom cells grew older. A lower fraction of dissolved ¹⁴C (12-23%) excreted by the copepods Acartia erythraea was detected in the colloidal phase compared to carcass (13-35%) and feces decomposition (21-34%). In contrast to carbon, a lower fraction of regenerated dissolved Cd (1-11%) and Zn (0-20%) from copepods and diatoms was consistently detected in the colloidal phases. Copepod excretion and carcass decomposition resulted in more colloidal Fe (51-91%) than diatom exudation (46-62% for Thalassiosira weissflogii, and 3-33% for T. pseudonana) and copepod feces decomposition (16-30%). Copepod (Calanus sinicus) grazing reduced the colloidal fraction of dissolved ¹⁴C, although a higher concentration of the diatom's (T. weissflogii) carbon was regenerated into the dissolved phase. The grazing of these copepods did not have any influence on the colloidal metal partitioning. The release of trace metals and carbon was enhanced by a higher density of copepod's grazing. Thus, different biological processes (grazing, excretion, exudation, and decomposition) may contribute differently to the production and dynamics of colloidal carbon and metals in planktonic systems.     

Spatial patterns of benthic diatoms,carbohydrates and mud on a tidal flat in the Ems-Dollard estuary

The chlorophyll a content and two operational fractions of carbohydrate (water extractable and EDTA extractable) were measured every three months during one year along transects on a tidal flat in the Ems-Dollard estuary (The Netherlands). Chlorophyll a was used as an indicator of microphytobenthos biomass, which was composed predominantly of epipelic diatoms. Both carbohydrate fractions correlated significantly with chlorophyll a. EDTA extractable carbohydrates were more resistant towards degradation than the water extractable fraction. During most of the year, concentrations of chlorophyll a and carbohydrates were low, but in June, high concentrations of up to 90 g chlorophyll a/g sediment were found in a narrow zone running parallel to the channel. Maximum concentrations of water extractable carbohydrates and EDTA extractable carbohydrates ranged between 800–1200 and 600–800 g/g sediment, respectively. The mud content was high (± 90%) at the margin of the tidal flat. This was not limited to the growth season of the diatoms, but was observed throughout the year. This indicated that the high mud content at the mudflat margin was mainly caused by hydrodynamic factors, and not by biostabilization. In June, exceptionally high diatom densities were found in sediment with a high mud content. There was only minor evidence that biostabilization by epipelic diatoms lead to a further increase in the mud content of the sediment.     

NMR structural study of fructans produced by Bacillus sp. 3B6, bacterium isolated in cloud water

Bacillus sp. 3B6, bacterium isolated from cloud water, was incubated on sucrose for exopolysaccharide production. Dialysis of the obtained mixture (MWCO 500) afforded dialyzate (DIM) and retentate (RIM). Both were separated by size exclusion chromatography. RIM afforded eight fractions: levan exopolysaccharide (EPS), fructooligosaccharides (FOSs) of levan and inulin types with different degrees of polymerization (dp 2–7) and monosaccharides fructose:glucose = 9:1. Levan was composed of two components with molecular mass ∼3500 and ∼100 kDa in the ratio 2.3:1. Disaccharide fraction contained difructose anhydride DFA IV. 1-Kestose, 6-kestose, and neokestose were identified as trisaccharides in the ratio 2:1:3. Fractions with dp 4–7 were mixtures of FOSs of levan (2,6-βFruf) and inulin (1,2-βFruf) type. DIM separation afforded two dominant fractions: monosaccharides with fructose: glucose ratio 1:3; disaccharide fraction contained sucrose only. DIM trisaccharide fraction contained 1-kestose, 6-kestose, and neokestose in the ratio1.5:1:2, penta and hexasaccharide fractions contained FOSs of levan type (2,6-βFruf) containing α-glucose. In the pentasaccharide fraction also the presence of a homopentasaccharide composed of 2,6-linked βFruf units only was identified. Nystose, inulin (1,2-βFruf) type, was identified as DIM tetrasaccharide. Identification of levan 2,6-βFruf and inulin 1,2-βFruf type oligosaccharides in the incubation medium suggests both levansucrase and inulosucrase enzymes activity in Bacillus sp. 3B6.     

Characterization of Microbial Mats from a Siliciclastic Tidal Flat (Bahía Blanca Estuary,Argentina)

Biofilms and microbial mats cover the tidal flats of the central zone of the Bahía Blanca estuary (Argentina), creating extensive layers. The objective of this study was to characterize the microphytobenthic communities in these biofilms and mats from sediment cores taken in March, June, September and December 2010. Microorganisms were identified and enumerated by microscopy, and their biomass (chlorophyll a, biovolume) quantified at two different stations in the lower supratidal zone, located ～210 m apart from each other (namely S1 and S2). Additionally, the colloidal carbohydrates produced by these microbial communities were quantified, together with physical parameters such as temperature, granulometry, moisture and organic matter content of the sediment layers that comprise a typical epibenthic mat. On the other hand, changes in biomass and colloidal carbohydrate content were studied through a half-tidal cycle (7 h). There were significant seasonal differences in microphytobenthic biovolume (P < 0.001) with a considerably lower biomass in summer, but no significant differences in microalgal biovolume between stations (P = 0.454). Cyanobacterial biomass (largely composed of the filamentous Microcoleus chthonoplastes) was dominant on all dates at both stations, followed by pennate diatoms. Chlorophyll a and colloidal carbohydrate contents in sediment presented a similar pattern to that of microalgal biovolume; with a 5-fold variation in chlorophyll a for S1 between consecutive sampling events on September and December. There were significant differences between sampling dates in colloidal carbohydrates (P < 0.001) with the lowest values recorded during fall and winter; conversely there were no significant differences between stations (P = 0.324). Silt was the dominant sediment fraction at S1 while sand dominated throughout the uppermost 20 mm at S2. Chlorophyll a contents did not show significant differences throughout a half-tidal cycle, likely the product of vertical migration along the section sampled. Conversely, the content of colloidal carbohydrates varied 5-fold, showing a significant (P < 0.001) and steady increase with time of exposure to air and pointing to the rapid metabolic rates of the community. In conclusion, the microphytobenthic community of the Bahía Blanca estuary presented marked seasonality in its biological parameters and overall physiognomy, also showing elevated metabolic rates when subject to tidal fluctuations.     

The production of extracellular carbohydrates by estuarine benthic diatoms: the effects of growth phase and light and dark treatment

Epipelic diatoms are important constituents of estuarine microphytobenthic biofilms. Field‐based investigations have shown that the production of carbohydrates by such taxa is ecologically important. However, limited information exists on the dynamics of carbohydrate production by individual species of epipelic diatoms. The production of low and high molecular weight extracellular carbohydrates in axenic cultures of five species of benthic estuarine diatoms, Cylindrotheca closterium (Ehrenberg), Navicula perminuta (Grun.) in Van Heurck, Nitzschia frustulum (Kütz.) Grunow, Nitzschia sigma (Kütz.) Grunow, and Surirella ovata (Kütz.) Grunow, were investigated. All species produced colloidal (water‐soluble) carbohydrates during growth, with maximal production occurring during stationary phase. During logarithmic growth, approximately 20% of extracellular carbohydrates consisted of polymeric material (extracellular polymeric substances [EPS]), but during stationary phase, EPS content increased to 34%–50%. Pyrolysis–mass spectrophotometry analysis showed differences in the composition of EPS produced during logarithmic and stationary phase. All species synthesized glucan as a storage carbohydrate, with maximum glucan accumulation during the transition from log to stationary phase. Short‐term labeling with ¹⁴C‐bicarbonate found that between 30 and 60% of photoassimilates were released as colloidal carbohydrate, with EPS consisting of approximately 16% of this colloidal fraction. When cells were placed in darkness, EPS production increased, and between 85 and 99% of extracellular carbohydrate produced was polymeric. Glucan reserves were utilized in dark conditions, with significant negative correlations between EPS and glucan for N. perminuta and S. ovata. Under dark conditions, cells continued to produce EPS for up to 3 days, although release of low molecular weight carbohydrates rapidly ceased when cells were dark treated. Three aspects of EPS production have been identified during this investigation: (1) production during rapid growth, which differs in composition from (2) EPS directly produced as a result of photosynthetic overflow during growth limiting conditions and (3) EPS produced for up to 3 days in the dark using intracellular storage reserves (glucans). The ecological implications of these patterns of production and utilization are discussed.     

Periodic acid-Schiff's reagent assay for carbohydrates in a microtiter plate format

Microtiter plate colorimetric assays are widely used for analysis of carbohydrates and glycoconjugates. However, mucins are often not easily detected, as they have low neutral sugar content. We have adapted and optimised the periodic acid–Schiff’s reagent (PAS) staining for microtiter plate assay by examining five factors: concentration and volume of periodic acid, oxidation time, volume of Schiff’s reagent, and color development time. This assay requires just 25 μl of sample, utilises standardised Schiff’s reagent, and has decreased assay time (140 min to completion). Seventeen monosaccharides (acidic, neutral, basic, phosphorylated, and deoxy) and four disaccharides were assessed. PAS-positive carbohydrates (amino, N-acetylamino, deoxy, and certain neutral monosaccharides, and sialic acids) responded linearly within a 10–100 nmol range approximately, which varied for each carbohydrate. The assay response for fetuin and porcine gastric mucin (PGM) was linear up to 150 μg (highest concentration tested), with no response from nonglycosylated protein. A lower response for asialofetuin was observed, but desialylated PGM preparations were similar or higher in response than their sialylated counterparts. The simplicity and low sample consumption of this method make it an excellent choice for screening or quantitation of chromatographic fractions containing carbohydrates and glycoconjugates, especially in the case of mucins.     

Cytokinins of sycamore spring sap

Summary The cytokinins present in the spring sap of Acer pseudoplatanus L. were investigated. Ribosyl-trans-zeatin, trans-zeatin and dihydrozeatin were isolated and identified by combined gas chromatography-mass spectrometry (GC-MS). A number of other cytokinin active fractions were observed. One of these was less polar than zeatin and did not behave as any known cytokinin. Two other fractions were more polar than ribosylzeatin and were unstable. A decomposition product of one of these was identified as ribosyl-trans-zeatin by GC-MS. The possible nature of the unstable compounds is discussed. Data on the changes in cytokinin activity of the various fractions during spring 1973 are presented and discussed.Abbreviations GLC gas-liquid chromatography - GG-MS combined gas chromatography-mass spectrometry - KE kinetin equivalents - TLC thin-layer chromatography - TMS trimethylsilyl - tRNA transfer RNA - i⁶ Ade 6-(3-methylbut-2-enylamino)-purine - i⁶ Ado 6-(3-methylbut-2-enylamino)-9--D-ribofuranosyl-purine     

Extracellular glycoconjugates produced by cyanobacterium Wollea saccata

In order to survive in a highly competitive environment, freshwater or marine phototrophic microorganisms have to develop defense strategies that result in a tremendous diversity of compounds from different metabolic pathways. Recent trends in drug research from natural sources have shown that algae and cyanobacteria are promising organisms to furnish novel biochemically active compounds. In this study, we have analysed the extracellular mucilaginous proteoglycan produced by fresh-water heterocytous filamentous cyanobacterium Wollea saccata, strain Hindák 2000/18. This mucilaginous material is an acidic proteoglycan containing 30% protein and 52% carbohydrates on the basis of fraction dry weight. The constituent sugars of the carbohydrate component include glucose, fucose, 3-O-methylfucose, xylose, galactose, 3-O-methylgalactose, mannose, rhamnose, arabinose and glucuronic acid. The extracellular proteoglycan has been separated into five fractions (WF1-WF5) by anion exchange chromatography. Individual polymeric fractions varied in protein (16-57%) and carbohydrate (31-66%) contents, and in the composition of constituent monosaccharides.     

Identification of ononitol and O-methyl-scyllo-inositol in pea root nodules

Ononitol (4-O-methyl-myo-inositol) and O-methyl-scyllo-inositol were identified in pea (Pisum sativum L.) root nodules formed by twoRhizobium leguminosarum strains. Ononitol was the major soluble carbohydrate in nodules formed by strain 1045 while O-methyl-scyllo-inositol and two unidentified components were dominant in the carbohydrate pattern of the nodules formed by strain 1 a. The cyclitols were also present in the denodulated roots, but to a much smaller extent; in the above-ground plant parts only traces were found. The identification of ononitol and O-methyl-scyllo-inositol was established by gas chromatography and gas chromatography-mass spectrometry utilizing trimethylsilyl- and acetyl-derivatives.Abbreviations GC-MS gas chromatography-mass spectroscopy - TLC thin-layer chromatography     

宁夏不同地区灵武长枣果实多糖的单糖成分分析

以宁夏4个不同地区(灵武、中宁、青铜峡、银川)成熟期的灵武长枣果实为研究对象,经水提醇沉法提取,采用DEAE-cellulose52和HW-55S分离纯化,并利用GC-MS法进行多糖的单糖组成分析。结果表明:多糖提取率最高的是灵武地区,达到1.795%;分离纯化后,4个地区的长枣多糖各得到1个中性(Ju-0)和3个酸性组分(Ju-1、Ju-2、Ju-3),其中Ju-2含量最高;GC-MS分析可知灵武长枣多糖含有阿拉伯糖、鼠李糖、核糖、岩藻糖、木糖、甘露糖、半乳糖、葡萄糖、葡萄糖醛酸、半乳糖醛酸10种单糖,不含果糖,以阿拉伯糖、核糖、半乳糖和2种糖醛酸为主,木糖含量最低。各地区多糖的单糖组成、含量各不相同,从各组分来看,四个地区多糖的Ju-0和Ju-1组分组成均以阿拉伯糖、核糖、半乳糖为主,四个地区多糖的组成差异主要在于Ju-2和Ju-3组分。从各地区单糖总量来看,灵武地区是阿拉伯糖含量最高,中宁、青铜峡、银川地区以葡萄糖醛酸含量为最高。     

Chemical Characteristics and Origin of Dissolved Organic Matter in the Yukon River

Monthly (or bi-weekly) water samples were collected from the Yukon River, one of the largest rivers in North America, at a station near the US Geological Survey Stevens Village hydrological station, Alaska from May to September 2002, to examine the quantity and quality of dissolved organic matter (DOM) and its seasonal variations. DOM was further size fractionated into high molecular weight (HMW or colloidal, 1 kDa–0.45 μm) and low molecular weight (LMW, <1 kDa) fractions. Dissolved organic carbon (DOC), colored dissolved organic matter (C-DOM) and total dissolved carbohydrate (TCHO) species were measured in the size fractionated DOM samples. Concentrations of DOC were as high as 2830 μmol-C l⁻¹ during the spring breakup in May and decreased significantly to 508–558 μmol-C l⁻¹ during open-water season (June–September). Within the DOC pool, up to 85% was in the colloidal fraction (1 kDa–0.45 μm) in early May. As DOC concentration decreased, this colloidal portion remained high (70–85% of the bulk DOC) throughout the sampling season. Concentrations of TCHO, including monosaccharides (MCHO) and polysaccharides (PCHO), varied from 722 μmol-C l⁻¹ in May to 129 μmol-C l⁻¹ in September, which comprised a fairly constant portion of bulk DOC (24±2%). Within the TCHO pool, the MCHO/TCHO ratio consistently increased from May to September. The C-DOM/DOM ratio and the size fractionated DOM increased from May to September, indicating that DOM draining into the Yukon River contained increased amounts of humified materials, likely related to a greater soil leaching efficiency in summer. The average composition of DOM was 76% pedogenic humic matter and 24% aquagenic CHO. Characteristics of soil-derived humic substances and low chlorophyll-a concentrations support a dominance of terrestrial DOM in Yukon River waters.     

Responses of estuarine intertidal microphytobenthic algal assemblages to enhanced ultraviolet B radiation

As a result of ozone depletion, ground doses of ultraviolet B (UVB) radiation in the mid latitudes of the Northern Hemisphere have increased since the 1980s, and current predictions indicate no possible alleviation until at least post 2020. Mudflats and sandflats are important coastal-zone habitats, and support extensive biofilms of benthic microalgae (microphytobenthos). In intertidal situations, these assemblages are exposed to high levels of UVB radiation during periods of tidal exposure. Exposure of intertidal biofilms dominated by epipelic (mud-inhabiting) diatoms to 0, 0.18 or 0.35 W m⁻² UVB radiation for between 4 and 10 days resulted in no significant decreases in the maximum PSII quantum efficiency (F_v/F_m) throughout diel exposure periods. Although the quantum efficiency of electron transport (F_q′/F_m′) showed significant reductions early in some experiments, the major response was an increase in F_q′/F_m′ in UVB exposed biofilms. This increase in F_q′/F_m′ was suggestive of a protective vertical migration down into the sediment. Single-cell and whole biofilm fluorescence imaging demonstrated, for the first time, that motile diatoms are able to detect UVB radiation independently of UVA or photosynthetically active radiation (PAR) and migrate rapidly down (within 15 min) into the sediments to avoid it. This behavioural acclimation mechanism appears to prevent significant accumulation of UVB induced damage to the algae. UVB exposure had no significant effect of biofilm photosynthesis (measured by ¹⁴C carbon fixation), but did alter organic carbon allocation patterns, with significantly less new carbon allocated to intracellular storage (glucan) and extracellular colloidal carbohydrate fractions. Significant reductions in the sediment standing stocks of chlorophyll a (Chl a), colloidal carbohydrates, extracellular polymeric substances (EPS) were seen after 7 days of UVB exposure. This study showed that marine intertidal benthic diatoms use a behavioural strategy to avoid exposure to UVB and that this response is effective as a short-term protection mechanism against UVB damage. However, altered carbon allocation patterns feed forward over time into changes in biofilm biomass and sediment carbohydrate dynamics. This suggests that continual long-term exposure to UVB may impact on sediment carbon cycling and trophic interactions and on the stabilization of sediments by microalgal biofilms through their production of extracellular carbohydrates.