Action of chitinase on spines of the diatom Thalassiosira fluviatilis

An electron microscopic study of the degradation of the chitin spines of the diatom Thalassiosira fluviatilis by Streptomyces chitinase revealed that only the apices of fibrils are broken down. The result suggests that the enzymolysis of crystalline chitinous structures may be rate-limited by the area of microfibril apices available, which varies with chitin source. A suspension of spines can be used as an assay for chitinase activity by monitoring the rate of loss of turbidity, and although this is not a sensitive assay, it does allow assessment of activity on a crystalline substrate.     

Cation-dependent changes in the thylakoid membrane appression of the diatom Thalassiosira pseudonana

Diatoms show a special organisation of their plastid membranes, such that their thylakoids span the entire plastid in bands of three. While in higher plants the interaction of the light harvesting complex II and photosystem II with divalent cations (especially Mg²⁺) was found to take part in the interplay of electrostatic attraction and repulsion in grana membrane appression, for diatoms the key players in maintaining proper membrane distances were not identified so far. In this work, we investigated the changes in the thylakoid architecture of Thalassiosira pseudonana in reaction to different salts by using circular dichroism and fluorescence spectroscopy in combination with other techniques. We show that divalent cations have an important influence on optimal pigment organisation and thus also on maintaining membrane appression. Thereby, monovalent cations are far less effective. The concentration needed is in a physiological range and fits well with the values obtained for higher plant grana stacking, despite the fact that strict protein segregation as seen in higher plant grana is missing.     

Influence of temperature and cell size on the division rate and chemical content of the diatom Chaetoceros curvisetum Cleve

Cell size and temperature influenced the division rate and chemical content of the diatom Chaetoceros curvisetum Cleve when grown at 15, 20 and 25°C in nutrient replete media. Cell-size dependent trends of division rate in individual clones changed with temperature in a complex fashion. Considerable interclonal variability in division rate within a restricted range of cell sizes was also found. Cellular levels of carbon, nitrogen, protein, chlorophyll a, and silicon were linearly related to cell size. Cellular levels of carbon and chlorophyll per unit volume and silicon per unit surface area changed with temperature. No temperature effect on cellular levels of nitrogen and protein was found.     

Effects of changes in ambient PAR and UV radiation on the nutritional quality of an Arctic diatom (Thalassiosira antarctica var. borealis)

Polyunsaturated fatty acids (PUFAs) are essential macromolecules that are synthesized by phytoplankton during spring bloom, and they play a key role in the Arctic food web. They are, however, considered to be sensitive to oxidation by UV radiation (280-400 nm). Changes in the food quality of primary producers may affect the transport of biomass and energy in the whole ecosystem. Using a common Arctic diatom, we looked at the effect of ambient and increased UV radiation on its nutritional quality, specifically, the fatty acid composition and elemental ratios. In May 2004, in the archipelago of Svalbard (79° N), a unialgal culture of Thalassiosira antarctica var. borealis was subjected to a 17-day experiment in outdoor aquaria. The diatoms were kept in semi-continuous culture (40 1) and exposed to three treatments with different levels of UV radiation: none (UV-shielded), ambient, and enhanced. Fatty acid composition, C:N:P ratios, photosynthetic pigment composition, optimum quantum yield of PSII, and cell numbers were analysed over the experimental period. An initial increase in PAR (photosynthetically active radiation, 400-700 nm) intensities profoundly affected the fatty acid composition and substantially inhibited the synthesis of PUFAs, but the relative amounts of PUFAs were not reduced by UV radiation. Enhanced UV radiation did, however, cause a significant reduction in optimum quantum yield of PSII and affected some fatty acids, mainly 18:0 and 16:1 n-7, during the first week of the experiment. Both ambient and enhanced UV radiation caused significantly lower C:P and N:P ratios. At the same time, these treatments elicited a higher relative content of the photoprotective pigments diadinoxanthin and diatoxanthin. After acclimation to the new light levels these effects faded off. Thus, brief periods with high light exposure may cause significant changes in photosynthetic activity and food quality, but the capacity for photo-acclimation seems high. The impact of UV radiation seems to be less important for food quality than that of PAR during a sudden rise in total light intensity.     

Effects of temperature, irradiance, and daylength on the marine diatom Leptocylindrus danicus Cleve. IV. Growth

Growth and dark respiration rates of the marine diatom Leptocylindrus danicus Cleve were measured in axenic batch culture under 49 combinations of temperature (5, 10, 15, 20°C), daylength(15:9, 12:12, 9:15 LD), and irradiance (at least four irradiances per daylength). Cell division rates exhibited a temperature-dependent daylength effect. Optimal temperatures occurred between 15 and 20°C. Both the initial slope () and the growth rate at light saturation (μ_max) were strongly influenced by temperature; increased five-fold and μ_max by an order of magnitude between 5 and 20°C. The compensation irradiance (I_c) was independent of temperature. μ_max was 2.7 div day⁻¹ at 20°C, 2.6 at 15°C, 1.1 at 10°C, and 0.3 at 5 °C. Cells grown under 15:9 and 12:12 LD exhibited similar growth-light curves at 20°C and at 15°C. μ_max of cells grown under 9:15 LD at these temperatures were substantially lower than μ_max under longer daylengths. Growth at 10 and 5°C was independent of daylength.

Dark respiration rates were a linear function of cell division rates at 10, 15, and 20°C, and support the concept that growth rate is dependent on dark respiration rate. These relationships were not influenced by daylength. A detectable relationship between dark respiration and growth at 5°C was not observed.

Photosynthesis and excretion showed temperature-dependent curvilinear relationships with growth rate, reflecting the lower saturation irradiance for growth compared to light saturation of photosynthesis and excretion. The relationship between Chl a-specific photosynthesis and growth was controlled by the C:Chl a ratio, which showed a positive correlation with cell division rate. At 15 and 20°C, light saturation of growth was associated with C:Chl a ratios of 40 to 60; at 5 and 10°C, cells growing at μ_max contained C:Chl a in ratios of 80 to 110.     

Activation of Thalassiosira pseudonana nadh: Nitrate reductase

NADH: nitrate reductase (NR) has been isolated in both active and inactive states, and both could be purified using blue-Sepharose. The state of activation of the enzyme depended on the presence or absence of agents such as cysteine or EDTA during the assay. When NR was assayed, the addition of activator before NADH led to maximum activity. Therefore, the reduced NR appeared to be inactivated during the assay in the absence of activator. Inactivation may have occurred via a mechanism similar to the inactivation of lipoamide dehydrogenase by trace metals, such as CU²⁺. The activation of NR by cysteine or EDTA was interpreted as protection of the reduced enzyme due to chelation of trace metals in the assay solution by the activators.     

Growth of three plankton diatom species in Oslofjord water in the absence of artificial chelators

Cultures of the plankton diatoms Skeletonema costatum (Greville) Cleve, Thalassiosira nordenskioeldii Cleve, and Lauderia annulata Cleve were grown in sea-water samples collected in the inner Oslofjord on 11 occasions between February, 1976 and January, 1977. The growth experiments were carried out in silicone-coated culture flasks, with nitrate, phosphate, and silicate added in excess, and under standardized conditions of a temperature of 10 °C and a saturating light intensity. Growth rates, as measured during exponential growth over a period of 5–6 days, were, with very few exceptions, close to the maximum rates obtained in a complete medium. The addition of ethylenediaminetetra-acetic acid (EDTA) to the water samples did not lead to improved growth, nor did EDTA have an effect on the initial lag phase. The lack of a seasonal variation in the observed growth responses was in marked contrast to the large fluctuations in the phytoplankton populations in the fjord. These findings give no support to the hypothesis that species successions among marine plankton diatoms are influenced by organic chelators or other extracellular substances produced by planktonic algae.     

The quaternary structure of pyruvate kinase type 1 from Escherichia coli at low nanomolar concentrations

Pyruvate kinase (PK) is the key control point of glycolysis—the biochemical pathway central to energy metabolism and the production of precursors used in biosynthesis. PK type 1 from Escherichia coli (Ec-PK1) is activated by both fructose-1,6-bisphosphate (FBP) and its substrate, phosphoenol pyruvate (PEP). To date, it has not been possible to determine whether the enzyme is tetrameric at the low concentrations (i.e. low nM range) used to study the steady-state kinetics, or assess whether its allosteric effectors alter the oligomeric state of the enzyme at these concentrations. Employing the new technique of analytical ultracentrifugation with fluorescence detection we have, for the first time, shown that the K_D^4–2 for Ec-PK1 is in the subnanomolar range, well below the concentrations used in kinetic studies. In addition, we show that, unlike some other PK isoenzymes, the modulation of oligomeric state by the allosteric effectors FBP and PEP does not occur at a concentration of 10 nM or above.     

Competition between Lemna minuta and Lemna minor at different nutrient concentrations

We investigated the differential responses of invasive alien Lemna minuta and native Lemna minor to nutrient loading as well as the mechanism of competition between the species. The role of nutrients, species identity, species influence in determining the outcome of competition between the species was estimated using the Relative Growth Rate Difference (RGRD) model. The two species differed in their response to nutrient loading. The native L. minor responded indifferently to nutrient loading. The species Relative Growth Rate (RGR) was 0.10 d⁻¹, 0.11 d⁻¹ and 0.09 d⁻¹ in high, medium and low nutrients, respectively. On the other hand, the invasive L. minuta responded opportunistically to high nutrient availability and had an RGR of 0.13 d⁻¹, 0.10 d⁻¹ and 0.08 d⁻¹ in high, medium and low nutrients, respectively. As a result, the invasive species was dominant in high nutrient availability but lost to the native species at low nutrient availability. The invader formed approximately 60% and less than 50% of the stand final total dry biomass in high and low nutrient availability, respectively. Species RGR were reduced by both intra- and interspecific competition but intraspecific effects were stronger than interspecific effects. On the overall, the species significantly differed in their constant RGR. These differences in RGR between the species (species identity) and the differential response to nutrient loading were the main determinant of change in final biomass composition of these species in mixture. Species influence (competition) only had a small influence on the outcome of competition between the species. The observed species response to nutrient loading could be targeted in management of the invasive species. Lowering nutrients can be proposed to reduce the impact of the invasive L. minuta.     

Growth and uptake kinetics of a facultatively oligotrophic bacterium at low nutrient concentrations

In oligotrophic waters, not only community structure but also physiological properties of heterotrophic bacteria are influenced by the concentration of organic matter.The relationship between growth rate of two facultatively oligotrophic strains ofAeromonas sp. No. 6 andFlavobacterium sp. M1 was studied in comparison with that of two eutrophic strains ofEscherichia coli 7020 andFlavobacterium sp. M2. These strains had two or three different substrate constants (Ks values) depending on substrate concentrations: Ks values for the two former were remarkably lower than those for the two latter. For instance, Ks value forAeromonas sp. No. 6 was about 8.9M when substrate concentration was greater than 53M and about 1.1M when substrate concentration was less man 53M. InE. coli the Ks value was about 260M at greater than 5600M and about 47M at less than 5600M substrate concentration.Uptake kinetics ofAeromonas sp. grown in a medium containing 2.7 mM glutamate (H-cell) and 0.11M glutamate (L-cell) have been determined for the intact cells. H-cell had two distinct values of Km for glutamate assimilation and respiration, and L-cell had three distinct values of Km for glutamate assimilation and respiration: In H-cell Km of assimilation was 2.8×10^–7 M and 1.5×10^–4 M, and Km of respiration was 2.3×10^–7 M and 1.7×10^–4 M; in L-cell Km of assimilation was 7.4×10^–8 M, 8.3×10^–6 M, and 1.3×10^–4 M, and Km of respiration was 2.5×10^–7, 8.9×10^–6M, and 1.7×10^–4 M. More than 60% of glutamate taken up by the H- and L-cells was respired when the substrate concentration was less than 10^–6 M, although at greater than 10^–6 M, 50% and 30% of glutamate was respired by H-cells and L-cells, respectively. These results suggest that the facultatively oligotrophic bacteria grow with high efficiency in environments with extremely low nutrient concentration, such as oligotrophic waters of lakes and ocean, as compared with in their growth in conditions of high nutrient concentraton, such as nutrient broth.     

Growth and stability of ethene-utilizing bacteria on compost at very low substrate concentrations

Ethene-utilising micro-organisms on compost may be applied in a packed bed to scrub the plant hormone ethene from air. Ethene at the concentrations tested (50–200 vpm) supported growth of micro-organisms present in compost and also of ethene-grown Mycobacterium E3 cells immobilised on compost.     

Uptake of glycollate by Skeletonema costatum (Grev.) Cleve in bacterized culture

Glycollic acid, supplied at a concentration of 1 mg l^?1, increased the relative growth rate of Skeletonema costatum (Grev.) Cleve growing in bacterized culture at limiting light intensities. There was little or no such effect at intensities approaching saturation. The presence in the medium of alumina, an adsorbent for glycollate, prolonged the lag phase, the cells remaining viable for up to 5 days. Uptake of glycollate was not appreciably affected by the bicarbonate concentration of the medium. After 3 h, 80–92% of the glycollate carbon assimilated was found in the alcohol and benzene insoluble fraction of the cells. This is in agreement with the supposition that glycollate carbon is as-similated directly by the diatom rather than after degradation by bacteria to carbon dioxide.     

Growth of Dioscorea deltoidea at high sugar concentrations

Dioscorea deltoidea cell suspension cultures were grown at initial sucrose concentrations of 35 to 200 g/L. The growth rates were similar (about 0.50 day^–1) with all of the initial sugar concentrations examined. The ratio of fresh weight to dry weight of cells was dependent on the initial sugar concentration, however, it remained fairly constant as long as the sugar was present in the growth medium. These results are different from results recently published, claiming that the growth rate of D. deltoidea cells is dependent on sugar concentration and the fresh weight to dry weight ratio increases throughout growth.     

Coexistence of two Cyclotella diatom species in the plankton of Lake Baikal

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The revised edition of a computerized plankton counter for plankton,periphyton and sediment diatom analyses

A revised computer counting program is presented with faster processing and more efficient features to enhance counting precision. The program is now capable of counting epilithic communities and sediment samples for diatom analysis. This version (2.3) is available for APPLE, IBM, TANDY-RADIO SHACK, and HYPERION systems using standard Basic commands. New improvements include, density and biovolume calculations, a Chi square transect test for randomness, D'Agostino's test for a normal taxa distribution and a three file output with raw and processed data.     

Sterols and fatty acids of the marine diatom biddulphia sinensis

Nine sterols, most showing Δ⁵- or Δ^5,22-unsaturation, were identified in the marine diatom Biddulphia sinensis. One sterol, cholesta-5,22E-dien-3β-ol, comprised 70–80% of the total sterols which is the first such predominance noted in a diatom. The only Δ⁷-sterol detected was cholest-7-en-3β-ol and this was a very minor component. A sterol showing unusual side-chain alkylation,23,24-dimethylcholesta-5,22E-dien-3β-ol, was identified for the first time in a diatom. Total fatty acids exhibited a predominance of Δ⁹- 16:1, 14:0, 20:5 and 16:0, typical of diatoms, although the proportions of these acids were found to vary with culture maturity. n-Heneicosahexaene was the major hydrocarbon together with a small amount of squalene.