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PubMed2002

The flavo-enzyme xanthine oxidase is under circadian control in the marine alga Gonyaulax.

The activity of xanthine oxidoreductases (xanthine oxidase, XO, EC 1.2.3.2 and xanthine dehydrogenase, XDH, EC 1.1.1.204) in partially purified extracts of Gonyaulax polyedra was measured over 24 h both in a light:dark cycle and in constant light. This is the first demonstration of xanthine oxidoreductase in a unicellular alga. The activity of the O2-dependent form (XO) was found to be 15 times higher in light than in darkness. The same time-of-day specific differences persisted in constant light, demonstrating a control of XO by the circadian clock. In contrast, the activity of the NAD-dependent form (XDH) is not under circadian control. Because pharmacological inhibition of XO also blocks the effect of blue light on the Gonyaulax circadian clock, the possible relationship between XO and light reception in this unicellular alga will be discussed.

Animals↗
PubMed2000

The unicellular green alga Chlamydomonas reinhardtii as an experimental system to study chloroplast RNA metabolism.

Chloroplasts are typical organelles of photoautotrophic eukaryotic cells which drive a variety of functions, including photosynthesis. For many years the unicellular green alga Chlamydomonas reinhardtii has served as an experimental organism for studying photosynthetic processes. The recent development of molecular tools for this organism together with efficient methods of genetic analysis and the availability of many photosynthesis mutants has now made this alga a powerful model system for the analysis of chloroplast biogenesis. For example, techniques have been developed to transfer recombinant DNA into both the nuclear and the chloroplast genome. This allows both complementation tests and analyses of gene functions in vivo. Moreover, site-specific DNA recombinations in the chloroplast allow targeted gene disruption experiments which enable a "reverse genetics" to be performed. The potential of the algal system for the study of chloroplast biogenesis is illustrated in this review by the description of regulatory systems of gene expression involved in organelle biogenesis. One example concerns the regulation of trans-splicing of chloroplast mRNAs, a process which is controlled by both multiple nuclear- and chloroplast-encoded factors. The second example involves the stabilization of chloroplast mRNAs. The available data lead us predict distinct RNA elements, which interact with trans-acting factors to protect the RNA against nucleolytic attacks.

Animals↗
PubMed1996

Synergistic iron reduction and citrate dissimilation by Shewanella alga and Aeromonas veronii

Two bacterial isolates from Great Bay Estuary, New Hampshire, in co-culture carried out anaerobic dissimilation of citric acid with Fe(III) as the terminal electron acceptor. Neither isolate oxidized citrate with Fe(III) anaerobically in axenic culture. The Fe(III) reducer, Shewanella alga strain BrY, did not grow anaerobically with citrate as an energy source. The citrate utilizer, Aeromonas veronii, did not reduce iron axenically with a variety of electron donors including citrate. The onset of iron reduction by the co-culture occurred after initiation of citrate dissimilation and just prior to initiation of growth by either organism (as measured by viable plate counts). Anaerobic culture growth rates and final cell densities of each bacterial strain were greater in co-culture than in axenic cultures. By 48 h of growth, the co-culture had consumed 27 mM citrate as compared with 12 mM dissimilated by the axenic culture of A. veronii. By 48 h the co-culture produced half as much formate (6 mM) and twice as much acetate (40 mM) as did A. veronii grown axenically (12 mM and 20 mM, respectively). Formate produced from citrate by A. veronii appeared to have supported growth and Fe(III) reduction by S. alga.Although not obligatory, nutrient coupling between these two organisms illustrates that fermentative (A. veronii-type) organisms can convert organic compounds such as citrate to those used as substrates by dissimilatory Fe(III) reducers, including S. alga. This synergism broadens the range of substrates available for iron reduction, stimulates the extent and rate of organic electron donor degradation (and that of iron reduction) and enhances the growth of each participant.

Journal Article↗
PubMed2003

Total arsenic content of nine species of Antarctic macro algae as determined by electrothermal atomic absorption spectrometry.

Total arsenic in nine species Antarctic macro algae has been measured, by electrothermal atomic absorption spectrometry using a Pd/Mg(NO3)2 matrix modifier, to determine their capacity to accumulate the element. Macro algae were collected in February during the 2000 austral summer season at Jubany Station (Argentinean scientific station) around Potter Cove, King George Island. An optimized two-step microwave (MW) program was used to digest the samples. Dried samples were treated with HNO3, H2O2, and HF, left overnight, then subjected to the first MW cycle. After cooling HNO3 and HClO4 were added and samples were subjected to the second MW cycle of digestion treatment. The effect of power and time on As recovery was examined. The analytical features of the method were: detection limit, 0.24 microg g(-1) (dry mass); precision (RSD), 4.2-5.7%; recovery 91-105%. A wide range of As-retention capacity (41.0-447 microg g(-1) dry mass) was observed among the different species. The highest levels of As were found in Phaeurus antarcticus (447 microg g(-1) dry mass). This organism satisfies several prerequisites to be considered for consideration as a biomonitor in future studies.

Journal Article↗
PubMed2000

Vanadium in marine mussels and algae.

A method is presented which is sensitive enough for the determination of vanadium (V) in marine organisms such as mussels and algae. It was sufficiently checked by a reference material and it was applied to V determination in blue mussels and brown algae from the German Bight.

Animals↗
PubMed1996

Chemometrical approaches to evaluate analytical data from aquatic macrophytes and marine algae.

The possibility of using aquatic macrophytes and algae as bioindicators of water pollution is investigated. The use of cluster analysis enables the treatment of the analytical data in a global way, revealing hidden clusters of similar behaviour between various biological species or sampling sites. A great number of species was collected from four rivers and fourteen coastal sites in Greece and were analysed for zinc, copper, cadmium, lead and manganese. The analytical data obtained were treated by hierarchical clustering, for marine algae and aquatic macrophytes separately.

Journal Article↗
PubMed2003

Gating of Cl- currents in protoplasts from the marine alga Valonia utricularis depends on the transmembrane Cl- gradient and is affected by enzymatic cell wall degradation.

The electrical properties of protoplasts of the turgor pressure-regulating giant marine alga Valonia utricularis were investigated by using the patch-clamp technique. In the whole-cell configuration, large inward currents were elicited by negative-going voltage pulses. The time-dependent component was predominantly carried by Cl-, as revealed by 'tail current' analysis. When experiments were performed on protoplasts directly after mechanical release from the 'mother cell', small outward currents were additionally observed at membrane voltages more positive than ECl-. These outward currents disappeared to a large extent after treatment of the protoplasts with a mixture of cell wall-degrading enzymes. Plots of the chord conductance versus the clamped membrane voltage revealed that enzymatic treatment affected the gating properties. By fitting Boltzmann distributions to the data, a midpoint potential of + 5 +/- 5 mV (n = 7) was obtained in symmetrical Cl- solutions for mechanically released protoplasts. In contrast, protoplasts treated additionally with enzymes exhibited a midpoint potential of -13 +/- 5 mV (n = 8). By varying the external and internal Cl- concentration, gating was also shown to depend on the Cl- gradient across the plasmalemma both in enzymatically treated and untreated protoplasts. Plotting of the midpoint potential against the Nernst potential of Cl- rendered a slope less than 1 (0.70 and 0.64, respectively) indicating that gating did not strictly depend on the electrochemical Cl- gradient. The voltage- and Cl--dependence as well as inhibition experiments with 4,4'-diisothiocyanatostilbene-2,2'-disulfonic acid (DIDS) suggested that the Cl- conductance of the membrane is dominated by the Valonia Anion Channel 1 (VAC1) described by Heidecker, M., Wegner, L.H., Zimmermann, U. 1999: A patch-clamp study of ion channels in proto-plasts prepared from the marine alga Valonia utricularis. J. Membrane Biol. 172:235-247. The relevance of the findings for membrane potential control and turgor regulation in V. utricularis as well as the general implications of the data for electrophysiological work on protoplasts (that are usually obtained by enzymatic digestion of plant tissue) are discussed.

Cells, Cultured↗
PubMed1999

Determination of the individual electrical and transport properties of the plasmalemma and the tonoplast of the giant marine alga Ventricaria ventricosa by means of the integrated perfusion/charge-pulse technique: evidence for a multifolded tonoplast.

The charge-pulse relaxation spectrum of nonperfused and perfused (turgescent) cells of the giant marine alga Ventricaria ventricosa showed two main exponential decays with time constants of approximately 0.1 msec and 10 msec, respectively, when the cells were bathed in artificial sea water (pH 8). Variation of the external pH did not change the relaxation pattern (in contrast to other giant marine algae). Addition of nystatin (a membrane-impermeable and pore-forming antibiotic) to the vacuolar perfusion solution resulted in the disappearance of the slow exponential, whereas external nystatin decreased dramatically the time constant of the fast one. This indicated (by analogy to corresponding experiments with Valonia utricularis, J. Wang, I. Spiess, C. Ryser, U. Zimmermann, J. Membrane Biol. 157: 311-321, 1997) that the fast relaxation must be assigned to the RC-properties of the plasmalemma and the slow one to those of the tonoplast. Consistent with this, external variation of [K+]o or of [Cl-]o as well as external addition of K+- or Cl--channel/carrier inhibitors (TEA, Ba2+, DIDS) affected only the fast relaxation, but not the slow one. In contrast, addition of these inhibitors to the vacuolar perfusion solution had no measurable effect on the charge-pulse relaxation spectrum. The analysis of the data in terms of the "two membrane model" showed that K+- and (to a smaller extent) Cl--conducting elements dominated the plasmalemma conductance. The analysis of the charge-pulse relaxation spectra also yielded the following area-specific data for the capacitance and the conductance for the plasmalemma and tonoplast (by assuming that both membranes have a planar surface): (plasmalemma) Cp = 0.82 * 10(-2) F m-2, Rp = 1.69 * 10(-2) Omega m2, Gp = 5.9 * 10(4) mS m-2, (tonoplast) Ct = 7. 1 * 10(-2) F m-2, Rt = 14.9 * 10(-2) Omega m2 and Gt = 0.67 * 10(4) mS m-2. The electrical data for the tonoplast show that (in contrast to the literature) the area-specific membrane resistance of the tonoplast of these marine giant algal cells is apparently very high as reported already for V. utricularis. The exceptionally high value of the area-specific capacitance could be explained - among other interpretations - by assuming a 9-fold enlargement of the tonoplast surface. The hypothesis of a multifolded tonoplast was supported by transmission electronmicroscopy of cells fixed under maintenance of turgor pressure and of the electrical parameters of the membranes. This finding indicates that the tonoplast of this species exhibited a sponge-like appearance. Taking this result into account, it can be easily shown that the tonoplast exhibits a high-resistance (1.1 Omega m2). Vacuolar membrane potential measurements (performed in parallel with charge-pulse relaxation studies) showed that the potential difference across the plasmalemma was mainly controlled by the external K+-concentration which suggested that the resting membrane potential of the plasmalemma is largely a K+-diffusion potential. After permeabilization of the tonoplast with nystatin the potential of the intact membrane barrier dropped from about slightly negative or positive (-5.1 to +18 mV, n = 13) to negative values (-15 up to -68 mV; n = 8). This indicated that the cytoplasm of V. ventricosa was apparently negatively charged relative to the external medium. Permeabilization of the plasmalemma by addition of external nystatin resulted generally in an increase in the potential to slightly more positive values (-0.8 to +4.3 mV; n = 5), indicating that the vacuole is positively charged relative to the cytoplasm. These findings apparently end the long-term debate about the electrical properties of V. ventricosa. The results presented here support the findings of Davis (Plant Physiol. 67: 825-831, 1981), but are contrary to the results of Lainson and Field (J. Membrane Biol. 29: 81-94, 1976).

Action Potentials↗
PubMed2003

A novel family of unconventional actins in volvocalean algae.

The unicellular green alga Chlamydomonas reinhardtii has two actin genes, one encoding a conventional actin (90% amino acid identity with mammalian actin), the other a highly divergent actin (64% identity) named novel actin-like protein (NAP). To see whether the presence of conventional and unconventional actins in a single organism is unique to C. reinhardtii, we searched for genomic sequences related to the NAP sequence in several other species of volvocalean algae. Here we show that Chlamydomonas moewusii and Volvox carteri also have, in addition to a conventional actin, an unconventional actin similar to the C. reinhardtii NAP. Analyses of the deduced protein sequences indicated that the NAP homologues form a distinct group derived from conventional actin.

Actins↗
PubMed2004

Lateral transfer and recompartmentalization of Calvin cycle enzymes of plants and algae.

Certain Calvin cycle enzymes also function in glycolysis or gluconeogenisis, thus photosynthetic eukaryotes would be predicted to have ancestrally possessed cytosolic homologues of these enzymes derived from the eukaryotic host and plastid homologues from the cyanobacterial endosymbiont. In practice, the evolutionary histories of these enzymes are often more complex. Focusing on eukaryotes with secondary plastids, we have examined the evolution of four such genes: class I and II fructose bisphosphate aldolase (FBA), sedoheptulose bisphosphatase (SBPase), and fructose bisphosphatase (FBPase). We show that previously observed distributions of plastid and cytosolic homologues are not always found in algae with secondary plastids: there is evidence for multiple events of both lateral gene transfer and retargeting to a new cellular compartment for both cytosolic and plastid enzymes of plants and algae. In particular, we show that a clade of class II FBAs spans a greater diversity of eukaryotes that previously recognized and contains both plastid-targeted (Phaeodactylum, Odontella) and cytosolic (ascomycetes, oomycetes, Euglena, and Bigelowiella) forms. Lateral transfer events also gave rise to a subset of plant cytosolic FBA, as well as cytosolic FBPase in Toxoplasma and other coccidian apicomplexa. In contrast, it has recently been suggested that the Trypanosoma FBA and SBPase are derived from a plastid, however, greater taxonomic sampling shows that these enzymes provide no evidence for a plastid-containing ancestor of Trypanosoma. Altogether, the evolutionary histories of the FBA and SBPase/FBPase gene families are complex, including extensive paralogy, lateral transfer, and retargeting between cellular compartments.

Eukaryota↗
PubMed2004

Procedures for determining the pesticide sensitivity of indigenous soil algae: a possible bioindicator of soil contamination?

Soil algae are present in all kinds of soils, both arable and virgin, in large amounts and in great diversity. These photosynthetic microorganisms, which are concentrated in the top few centimeters of the soil profile, are organized in a community structure that varies depending on soil type, farming method and pesticide application. We first tested several extraction and conservation methods for soil algae, and a 14C incorporation procedure to evaluate the photosynthetic activity of these micro-organisms. In a second study, we assessed the sensitivity to atrazine of soil microalgae from two corn fields managed by different agricultural practices (conventional vs. organic). Changes in indigenous diatom communities were monitored, together with photosynthetic tests performed on the whole algal community. Comparison of the data for the treated field with those for the reference, untreated organic cornfield showed that previous atrazine application in the conventional cornfield had changed the species composition of the soil diatom communities. Short-term ecotoxicological tests, using photosynthetic activity as endpoint, also showed that the communities that had developed under pesticide stress were more tolerant to further atrazine application than the control communities. The stress caused by major environmental disturbances, such as ground dressing, was not sufficient to mask the difference between the two crops, suggesting that telluric microalgae could be used as indicators of xenobiotic contamination in soils. These initial findings about using soil microalgal communities as bioindicators are promising. In addition, their photosynthetic activity, which reflects their sensitivity to xenobiotic compounds, seems to be a relevant bioindicator of soil contamination.

Biomarkers↗
PubMed1998

Liquid-gas partitioning of the gasoline oxygenate methyl tert-butyl ether (MTBE) under laboratory conditions and its effect on growth of selected algae.

The partitioning of the widely used gasoline additive methyl tert-butyl ether (MTBE) between liquid growth media and gaseous phase was measured daily under laboratory conditions to determine how closely dissolved MTBE concentrations matched nominal concentrations. Total (gaseous and dissolved) MTBE averaged across 6 days for 29.6, 503.2, and 1005.7 mg L-1 MTBE treatments were 89.9, 90.3, and 73.0% of nominal, respectively, and mean dissolved MTBE in these same treatments were 74.6, 73.8, and 69.6% of total MTBE, respectively. This suggests that dissolved MTBE concentrations can vary substantially from nominal. The effect of MTBE on the growth of selected algae was also evaluated under laboratory conditions. Three unicellular algae, Selenastrum capricornutum (Chlorophyta), Navicula pelliculosa (Bacillariophyta), and Synechococcus leopoliensis (= Anacystic nidulans, Cyanophyta = Cyanobacteria), representative of three taxonomic groups, were used as test organisms. Toxicity tests were acute and increase in cell number was used as an indicator of growth. Algal species were exposed by injection of MTBE into sealed vessels containing defined liquid growth media. The growth of N. pelliculosa and S. leopoliensis was negatively affected at nominal 2400 mg L-1 MTBE, whereas the growth of S. capricornutum was negatively affected at nominal 4800 mg L-1 MTBE and positively affected at nominal 600 mg L-1 MTBE. The differential sensitivity of the growth of these representative species suggests that MTBE may alter algal community composition in the natural environment.

Air Pollutants↗
PubMed1998

Influence of Dispersants on the Bioavailability of Naphthalene from the Water-Accommodated Fraction Crude Oil to the Golden-Brown Algae, Isochrysis galbana

The golden-brown algae Isochrysis galbana, a primary producer, was used to determine the influence of the chemical dispersing agent, Corexit 9527((R)), on the bioavailability of naphthalene. Cells were exposed to laboratory preparations of either the water-accommodated fraction (WAF) of Prudhoe Bay crude oil (PBCO) or a dispersed oil (DO) mixture of PBCO and Corexit 9527 spiked with [U-14C]naphthalene. Uptake was determined by the amount of algae-associated [14C]. High-pressure liquid chromatography (HPLC) co-chromatography was used to fractionate and identify metabolic products. A 24-h bioaccumulation factor (BAF) was calculated in the absence of steady state. The presence of Corexit 9527, had significant influence (p = 0.001) on the uptake of naphthalene, but no significant effect on the 24-h BAF (BAF: 168 and 180 from WAF and DO, respectively), or metabolic fate of naphthalene in I. galbana. Results of this research indicate that dispersants have the potential to increase organismal exposure to certain petroleum hydrocarbons without increasing their aqueous concentration.

Journal Article↗
PubMed2006

Microbial communities on glacier surfaces in Svalbard: impact of physical and chemical properties on abundance and structure of cyanobacteria and algae.

Microbial communities occurring in three types of supraglacial habitats--cryoconite holes, medial moraines, and supraglacial kames--at several glaciers in the Arctic archipelago of Svalbard were investigated. Abundance, biovolume, and community structure were evaluated by using epifluorescence microscopy and culturing methods. Particular emphasis was laid on distinctions in the chemical and physical properties of the supraglacial habitats and their relation to the microbial communities, and quantitative multivariate analyses were used to assess potential relationships. Varying pH (4.8 in cryoconite; 8.5 in a moraine) and texture (the proportion of coarse fraction 2% of dry weight in cryoconite; 99% dw in a kame) were found, and rather low concentrations of organic matter (0.3% of dry weight in a kame; 22% dw in cryoconite) and nutrients (nitrogen up to 0.4% dw, phosphorus up to 0.8% dw) were determined in the samples. In cryoconite sediment, the highest numbers of bacteria, cyanobacteria, and algae were found, whereas relatively low microbial abundances were recorded in moraines and kames. Cyanobacterial cells were significantly more abundant than microalgal ones in cryoconite and supraglacial kames. Different species of the cyanobacterial genus Leptolyngbya were by far the most represented in all samples, and cyanobacteria of the genera Phormidium and Nostoc prevailed in cultures isolated from cryoconite samples. These species are considered opportunistic organisms with wide ecological valency and strong colonizing potential rather than glacial specialists. Statistical analyses suggest that fine sediment with higher water content is the most suitable condition for bacteria, cyanobacteria, and algae. Also, a positive impact of lower pH on microbial growth was found. The fate of a microbial cell deposited on the glacier surface seems therefore predetermined by the physical and chemical factors such as texture of sediment and water content rather than spatial factors or the origin of sediment.

Cyanobacteria↗
PubMed2002

Fed-batch cultivation of the docosahexaenoic-acid-producing marine alga Crypthecodinium cohnii on ethanol.

The heterotrophic marine microalga Crypthecodinium cohnii produces docosahexaenoic acid (DHA), a polyunsaturated fatty acid with food and pharmaceutical applications. So far, DHA production has been studied with glucose and acetic acid as carbon sources. This study investigates the potential of ethanol as an alternative carbon source for DHA production by C. cohnii. In shake-flask cultures, the alga was able to grow on ethanol. The specific growth rate was optimal with 5 g l(-1) ethanol and growth did not occur at 0 g l(-1) and above 15 g l(-1). By contrast, in fed-batch cultivations with a controlled feed of pure ethanol, cumulative ethanol addition could be much higher than 15 g l(-1), thus enabling a high final cell density and DHA production. In a representative fed-batch cultivation of C. cohnii with pure ethanol as feed, 83 g dry biomass l(-1), 35 g total lipid l(-1) and 11.7 g DHA l(-1) were produced in 220 h. The overall volumetric productivity of DHA was 53 mg l(-1 )h(-1), which is the highest value reported so far for this alga.

Animals↗
PubMed2000

Biosorption of heavy metals by marine algae.

The ability of four different algae (three brown and one red) that have not been previously studied to adsorb Cr(3+), Co(2+), Ni(2+), Cu(2+), and Cd(2+) ions was investigated. The metal uptake was dependent on the type of biosorbent, with different accumulation affinities towards the tested elements. The HCl-treated biomass decreased the metal biosorptive capacity particularly in the case of Cr(3) adsorption with Laurencia obtusa. The extent of uptake of the different metals with the tested algae was assessed under different conditions such as pH, time of algal residence in solution with the metal, and concentration of algal biomass. The rate of uptake of the different metals was very fast in the first 2 h; thereafter the increase in metal uptake was insignificant. The amount of the metal uptake (5-15 mg range) increased steeply by increasing the weight of the biomass. An exception was L. obtusa, where a parallel increase of the uptake of different metals was observed on increasing the algal mass from 5 to 50 mg.

Biomass↗
PubMed2001

Adhesion of the dissimilatory Fe(III)-reducing bacterium Shewanella alga BrY to crystalline Fe(III) oxides.

Shewanella alga BrY adhesion to hydrous ferric oxide, goethite, and hematite was examined. Adhesion to each oxide followed the Langmuir adsorption model. No correlation between adhesion and Fe(III) oxide surface area or crystallinity was observed. Zeta potential measurements suggested that electrostatic interactions do not influence S. alga BrY adhesion to these minerals. Cell adhesion does not appear to explain the recalcitrance of crystalline Fe(III) oxides to bacterial reduction.

Bacterial Adhesion↗
PubMed2003

Characterization of apcC, the nuclear gene for the phycobilisome core linker polypeptide L(c)(7.8) from the glaucocystophyte alga Cyanophora paradoxa. Import of the precursor into isolated cyanelles and integration of the mature protein into intact phycobilisomes.

Phycobilisomes are the complex and highly efficient light-harvesting antenna systems of cyanobacteria, glaucocystophyte algae and red algae. In the glaucocystophyte Cyanophora paradoxa, seven genes for (chromophoric) phycobilisome components are known thus far, which all reside on the cyanelle genome. Here, we report the sequence of apcC, specifying the precursor to the colorless polypeptide L(c)(7.8), the first core linker reported for a eukaryote. The precursor was efficiently imported in vitro into isolated cyanelles. Fractionation into thylakoid membranes and stroma and into intact phycobilisomes and soluble proteins, respectively, indicated a low but significant incorporation of the imported linker polypeptide into the phycobilisomes.

Algal Proteins↗