The uptake of amino acids by the cyanobacterium Planktothrix rubescens is stimulated by light at low irradiances

The rates of uptake of five amino acids - alanine, glutamate, glycine, leucine and serine - by axenic cultures of the cyanobacterium Planktothrix rubescens were measured over a range of irradiances using the 14C-labelled amino acids at the nanomolar concentrations observed in Lake Zürich. The rates in the light exceeded the dark rates by as much as two- to ninefold. The light-affinity constants for stimulation were similar, indicating a similar process for each of the five amino acids. The Ek (light saturation irradiance) for light stimulation was only 1 μmol m-2 s-1, less than the compensation point for photosynthesis and autotrophic growth, and much lower than the Ek for either process. The Ek for amino acid uptake was also less than the irradiance at which filaments obtain neutral buoyancy, which determines the depth at which they stratify and the irradiance they receive. This indicates that stimulation of amino acid uptake by light of low irradiances provides a mechanism for supplementing growth of filaments stratifying deep in the metalimnion, which, while able to grow at low irradiances, are often left with insufficient light to sustain them. Acetate uptake was also stimulated by light, but the kinetics differe

Photosynthetic activity and population dynamics of Amoebobacter purpureus in a meromictic saline lake

Abstract A dense population of the purple sulfur bacterium Amoebobacter purpureus in the chemocline of meromictic Mahoney Lake (British Columbia, Canada) underwent consistent changes in biomass over a two year study period. The integrated amount of bacteriochlorophyll reached maxima in August and declined markedly during early fall. Bacteriochlorophyll was only weakly correlated with the light intensity and water temperature in the chemocline. In the summer, bacterial photosynthesis was limited by sulfide availability. During this period the intracellular sulfur concentration of A. purpureus cells decreased. A minimum concentration was measured at the top of the bacterial layer in August, when specific photosynthetic rates of A. purpureus indicated that only 14% of the cells were photosynthetically active. With the exception of a time period between August and September, the specific growth rates calculated from CO2 fixation rates of A. purpureus were similar to growth rates calculated from actual biomass changes in the bacterial layer. Between August and September 86% of the A. purpureus biomass disappeared from the chemocline and were deposited on the littoral sediment of Mahoney Lake or degraded within the mixolimnion. This rise of cells to the lake surface was not mediated by an increase in the specific gas vesicle content which remained constant between April and November. The upwelling phenomenon was related to the low sulfur content of A. purpureus cells and a low resistance of surface water layers against vertical mixing by wind

Solid-state NMR evidence for inequivalent GvpA subunits in gas vesicles

Gas vesicles are organelles that provide buoyancy to the aquatic microorganisms that harbor them. The gas vesicle shell consists almost exclusively of the hydrophobic 70-residue gas vesicle protein A, arranged in an ordered array. Solid-state NMR spectra of intact collapsed gas vesicles from the cyanobacterium Anabaena flos-aquae show duplication of certain gas vesicle protein A resonances, indicating that specific sites experience at least two different local environments. Interpretation of these results in terms of an asymmetric dimer repeat unit can reconcile otherwise conflicting features of the primary, secondary, tertiary, and quaternary structures of the gas vesicle protein. In particular, the asymmetric dimer can explain how the hydrogen bonds in the β-sheet portion of the molecule can be oriented optimally for strength while promoting stabilizing aromatic and electrostatic side-chain interactions among highly conserved residues and creating a large hydrophobic surface suitable for preventing water condensation inside the vesicle.National Institutes of Health (U.S.) (Grant EB002175)National Institutes of Health (U.S.) (Grant EB003151)National Institutes of Health (U.S.) (Grant EB002026

The critical pressures of gas vesicles in Planktorhrix rubescens in relation tothe depth of winter mixing in Lake Zürich, Switzerland

The vertical distribution of the cyanobacterium Planktothrir (Oscillazoria) rubescens in Lake Zürich was investigated from March 1993 to June 1995 by collecting filaments on filters and measuring them by epifluorescence microscopy and computer image analysis. The initial population, which began to stratify in April, decreased by up to 99% by June. During the summer, the population peaked at depths of 8-15 m; it reached a maximum areal filament-volume concentration of -60 cm −3 of lake surface in early September and was then entrained in the deepening surface layer. It became mixed progressively deeper, to the lake bottom in the cold winter of 1993-94, but less completely in the milder winter of 1994-95. Most of the filaments remained viable during the winter. At the end of the mild winter of 1994-5, 70% of filaments in the water column retained buoyancy, but after the cold winter of 1996-7 only 22% were buoyant. Few remained buoyant below 80 m, where the hydrostatic pressure caused gas vesicle collapse. The proportion that remain buoyant decreases with the depth and duration of winter mixing, and increases with the critical collapse pressure (Pc) of the gas vesicles, which provide buoyancy. Strains of P.rubescens isolated from Lake Zürich differed in mean (Pc) of their gas vesicles, from 0.9 to 1.1 MPa, the highest values in freshwater cyanobacteria. Allowing for a turgor pressure of 0.2 MPa. these strains would remain buoyant at depths down to 70 and 90 m, respectively. Natural selection for gas vesicles of high (Pc) will operate by increasing the proportion of filaments that remain buoyant in the upper parts of the water column after circulation to various depths during the winter because only buoyant filaments will form the inoculum for the following seaso

The gas vesicles, buoyancy and vertical distribution of cyanobacteria in the Baltic Sea

The mean pressures required to collapse gas vesicles in turgid cells of cyanobacteria from the Baltic Sea were 0·91 MPa (9·1 bar) in Aphanizomenon flos-aquae, 0·83 MPa in Nodularia sp. collected from the main deep basins and 0·34 MPa in Nodularia from shallower coastal regions. The gas vesicles were strong enough to withstand the depth of winter mixing, down to the permanent halocline (60 m in the Bornholm Sea, 90 m in the Eastern Gotland Sea) or to the sea bottom (30 m or less in the shallow Arkona Sea and Mecklenburg Bight). The cyanobacteria had low cell turgor pressures, within the range 0·08–0·18 MPa. The colonies were highly buoyant: the Aphanizomenon colonies floated up at a mean velocity of 22 m per day and the Nodularia colonies at 36 m per day. The colonies remained floating when up to half of the gas vesicles had been collapsed. In summer the cyanobacteria were mostly restricted to the water above the thermocline and in calm conditions their concentration increased towards the top of the water column. A series of colony concentration profiles indicated that, following a deep mixing event, the population of colonies moved upward with a net velocity of 22 m per day, similar to the colony floating velocity. This demonstrated that the buoyancy provided by gas vesicles would give a selective advantage to populations of cyanobacteria by enabling them to float into the higher irradiance of the near-surface water

Buoyancy regulation and aggregate formation in Amoebobacter purpureus from Mahoney lake

Abstract The meromictic Mahoney Lake (British Columbia, Canada) contains an extremely dense layer of purple sulfur bacteria (Amoebobacter purpureus). The buoyant density of Amoebobacter cells grown in pure culture at saturating light intensity was significantly higher (1027–1034 kg m−3) than the density of lake water (1015 kg m−3). When stationary cultures were shifted to the dark, the gas-vesicle content increased by a factor of 9 and buoyant density decreased to 1002 kg m−3 within three days. A novel mechanism of cell aggregation was detected for the Mahoney Lake strain. Dense cell aggregates were formed after depletion of sulfide. Formation of aggregates was correlated with an increase in cell surface hydrophobicity. Cell aggregates could be disintegrated within less than 1 s by addition of sulfide or various thiol compounds. Mercaptanes with a branched structure in the vicinity of the terminal thiol group, compounds with esterified thiol groups (methylmercaptanes), reducing compounds lacking thiol groups and detergents did not influence aggregate stability. Cell aggregates disintegrated upon addition of urea or of proteinase K. Addition of various sugars had no effect on aggregation; this points to the absence of lectins. The results indicate that cell-to-cell adhesion in A, purpureus ML1 is mainly caused by a hydrophobic effect and includes a specific mechanism possibly mediated by a surface protein. Extrapolation of laboratory results to field conditions demonstrated that both regulation of buoyant density and formation of cell aggregates result in passive accumulation of cells at the chemocline and contribute to the narrow stratification of A. purpureus in Mahoney Lake

Development of image analysis techniques as a tool to detect and quantify morphological changes in anaerobic sludge : I. application to a granulation process

Image analysis techniques were developed and applied to quantify the process of anaerobic granulation in an expanded granular sludge blanket reactor (EGSB) fed with a synthetic substrate based on glucose [60-30% COD chemical oxygen demand)] and volatile fatty acids (40-70% COD) over 376 days. In a first operation period that lasted 177 days, the aggregation of dispersed sludge was quantitatively monitored through the recognition and quantification of aggregates and filaments. A parameter defined as the ratio between the filaments' length and the aggregates projected area (LfA) has proven to be sensitive to detect changes in the aggregation status of the anaerobic sludge. The aggregation time-defined as the moment when a balance between filaments' length and aggregates' size was establishe-was recognized through the LfA. The percentage of projected area of aggregates within three size ranges (0.01 -0.1 mm, 0.1 - 1 mm, and >1 mm, equivalent diameter) reflected the granular size spectrum during the aggregation process. When sudden increases on the upflow velocity and on the organic loading rate were applied to the previously formed granules, the developed image analysis techniques revealed to be good indicators of granular sludge stability, since they were sensitive to detected filaments release, fragmentation, and erosion that usually leads to washout. The specific methanogenic activities in the presence of acetate, propionate, butyrate, and H2/CO2 increased along the operation, particularly relevant was the sudden increase in the specific hydrogenophilic activity, immediately after the moment recognized as aggregation time.Instituto Cooperação Científica e Tecnológica Internacional (ICCTI), Ambassade de France in Portugal - Project 203 B4.Fundação para a Ciência e a Tecnologia (FCT) - SFRH/BD/3187/2000, PRAXIS XXI/BD/20325/99, POCTI/1999/CTA736524

Sinking properties of some phytoplankton shapes and the relation of form resistance to morphological diversity of plankton – an experimental study

Form resistance (Phi) is a dimensionless number expressing how much slower or faster a particle of any form sinks in a fluid medium than the sphere of equivalent volume. Form resistance factors of PVC models of phytoplankton sinking in glycerin were measured in a large aquarium (0.6 x 0.6 x 0.95 m). For cylindrical forms, a positive relationship was found between Phi and length/ width ratio. Coiling decreased Phi in filamentous forms. Form resistance of Asterionella colonies increased from single cells up to 6-celled colonies than remained nearly constant. For Fragilaria crotonensis chains, no such upper limit to Phi was observed in chains of up to 20 cells ( longer ones were not measured). The effect of symmetry on Phi was tested in 1 - 6-celled Asterionella colonies, having variable angles between the cells, and in Tetrastrum staurogeniaeforme coenobia, having different spine arrangements. In all cases, symmetric forms had considerably higher form resistance than asymmetric ones. However, for Pediastrum coenobia with symmetric/asymmetric fenestration, no difference was observed with respect to symmetry. Increasing number and length of spines on Tetrastrum coenobia substantially increased Phi. For a series of Staurastrum forms, a significant positive correlation was found between arm-length/cell-width ratio and Phi: protuberances increased form resistance. Flagellates (Rhodomonas, Gymnodinium) had a Phi 1. The highest value ( Phi = 8.1) was established for a 20-celled Fragilaria crotonensis chain. Possible origin of the so-called 'vital component' ( a factor that shows how much slower viable populations sink than morphologically similar senescent or dead ones) is discussed, as is the role of form resistance in evolution of high diversity of plankton morphologies

Control of primary productivity and the significance of photosynthetic bacteria in a meromictic kettle lake.

During 1986 planktonic primary production and controlling factors were investigated in a small (A0 = 11.8 · 103 m2, Zmax = 11.5 m) meromictic kettle lake (Mittlerer Buchensee). Annual phytoplankton productivity was estimated to ca 120 gC · m–2 · a–1 (1,42 tC · lake–1 · a–1). The marked thermal stratification of the lake led to irregular vertical distributions of chlorophylla concentrations (Chla) and, to a minor extent, of photosynthesis (Az). Between the depths of 0 to 6 m low Chla concentrations (< 7 mg · m–3) and comparatively high background light attenuation (kw = 0,525 m–1, 77% of total attenuation due to gelbstoff and abioseston) was found. As a consequence, light absorption by algae was low (mean value 17,4%) and self-shading was absent. Because of the small seasonal variation of Chla concentrations, no significant correlation between Chla and areal photosynthesis (A) was observed. Only in early summer (June–July) biomass appears to influence the vertical distribution of photosynthesis on a bigger scale. Around 8 m depth, low-light adapted algae and phototrophic bacteria formed dense layers. Due to low ambient irradiances, the contribution of these organisms to total primary productivity was small. Primary production and incident irradiance were significantly correlated with each other (r2 = 0.68). Although the maximum assimilation number (Popt) showed a clear dependence upon water temperature (Q10 = 2.31), the latter was of minor importance to areal photosynthesis