Carbon dioxide biofixation by Chlorella vulgaris at different CO2 concentrations and light intensities

In order to develop an effective CO₂ mitigation process using microalgae for potential industrial application, the growth and physiological activity of Chlorella vulgaris in photobioreactor cultures were studied. C. vulgaris was grown at two CO₂ concentrations (2 and 13% of CO₂ v/v) and at three incident light intensities (50, 120 and 180 μmol m^?2 s^?1) for 9 days. The measured specific growth rate was similar under all conditions tested but an increase in light intensity and CO₂ concentration affected the biomass and cell concentrations. Although carbon limitation was observed at 2% CO₂, similar cellular composition was measured in both conditions. Light limitation induced a net change in the growth behavior of C. vulgaris. Nitrogen limitation seemed to decrease the nitrogen quota of the cells and rise the intracellular carbon:nitrogen ratio. Exopolysaccharide production per cell appeared to be affected by light intensity. In order to avoid underestimation of the CO₂ biofixation rate of the microalgae, exopolysaccharide production was taken into account. The maximum CO₂ removal rate (0.98 g CO₂ L^?1 d^?1) and the highest biomass concentration (4.14 g DW L^?1) were determined at 13% (v/v) CO₂ and 180 μmol m^?2 s^?1. Our results show that C. vulgaris has a real potential for industrial CO₂ remediation.     

Phytoplankton response to light and internal phosphorus loading from sediment release

1. A 2 × 2 factorial design was employed to look at the influence of two levels of phosphorus (P; high and low) and two levels of light (high and low) and their interactions, on phytoplankton abundance, elemental tissue composition and community structure in two seasons (April and June) of 2005. 2. A novel feature of the experiment was the creation of high and low P levels by manipulating sediment core conditions in the laboratory. Sediment cores were incubated with their associated overlaying water column from four different sites in Mona Lake, Michigan, under anaerobic or aerobic conditions, respectively. 3. After 24 days, the water overlaying the sediment cores was collected and used as growth media for phytoplankton collected from Mona Lake. Phytoplankton communities were grown in the laboratory in the high or low P water, and subjected to high (250 μmol m^?2 s^?1) or low (10 μmol m^?2 s^?1) light for 9 (April) or 14 (June) days. 4. In the April experiment, high P treatments resulted in significantly higher chlorophyll‐a concentrations, significantly lower C : P ratios from two of the four sites, and greater dominance by Scenedesmus at all sites relative to low P treatments. High light generally led to higher chlorophyll‐a concentrations, higher C : P ratios and greater Scenedesmus and Fragilaria biovolume at all sites. A significant interaction was measured between P and light for chlorophyll‐a and Scenedesmus biovolume, suggesting the influence of P was more apparent at high light than at low light levels. 5. In the June experiment, high P increased ash‐free dry mass (AFDM), lowered C : P ratios and resulted in increased Pediastrum biovolume. High light levels led to greater chlorophyll‐a concentrations, AFDM and C : P ratios, as well as increased biovolumes of Scendesmus, Pediastrum and Fragilaria. A significant interaction was found between P and light for all three taxa, as the positive influence of P was more pronounced at high light levels. 6. The results of our study demonstrate that sediment‐derived P stimulates phytoplankton growth, but that its effect on phytoplankton dynamics is modulated by other factors, such as light.     

Do phytoplankton communities correctly track trophic changes? An assessment using directly measured and palaeolimnological data

1. Measurements of total phosphorus (TP) concentrations since 1975 and a 50‐year time series of phytoplankton biovolume and species composition from Lake Mondsee (Austria) were combined with palaeolimnological information on diatom composition and reconstructed TP‐levels to describe the response of phytoplankton communities to changing nutrient conditions. 2. Four phases were identified in the long‐term record. Phase I was the pre‐eutrophication period characterised by TP‐levels of about 6 μg L^?1 and diatom dominance. Phase II began in 1966 with an increase in TP concentration followed by the invasion of Planktothrix rubescens in 1968, characterising mesotrophic conditions. Phase III, from 1976 to 1979, had the highest annual mean TP concentrations (up to 36 μg L^?1) and phytoplankton biovolumes (3.57 mm³ L^?1), although reductions in external nutrient loading started in 1974. Phases II and III saw an expansion of species characteristic of higher nutrient levels as reflected in the diatom stratigraphy. Oligotrophication (phase IV) began in 1980 when annual average TP concentration, Secchi depth and algal biovolume began to decline, accompanied by increasing concentrations of soluble reactive silica. 3. The period from 1981 to 1986 was characterised by asynchronous trends. Annual mean and maximum total phytoplankton biovolume initially continued to increase after TP concentration began to decline. Reductions in phytoplankton biovolume were delayed by about 5 years. Several phytoplankton species differed in the timing of their responses to changing nutrient conditions. For example, while P. rubescens declined concomitantly with the decline in TP concentration, other species indicative of higher phosphorus concentrations, such as Tabellaria flocculosa var. asterionelloides, tended to increase further. 4. These data therefore do not support the hypotheses that a reduction in TP concentration is accompanied by (i) an immediate decline in total phytoplankton biovolume and (ii) persistence of the species composition characterising the phytoplankton community before nutrient reduction.     

THE INFLUENCE OF SUBMERGED MACROPHYTES ON SEDIMENTARY DIATOM ASSEMBLAGES1

Submerged macrophytes are a central component of lake ecosystems; however, little is known regarding their long‐term response to environmental change. We have examined the potential of diatoms as indicators of past macrophyte biomass. We first sampled periphyton to determine whether habitat was a predictor of diatom assemblage. We then sampled 41 lakes in Quebec, Canada, to evaluate whether whole‐lake submerged macrophyte biomass (BiomEpiV) influenced surface sediment diatom assemblages. A multivariate regression tree (MRT) was used to construct a semiquantitative model to reconstruct past macrophyte biomass. We determined that periphytic diatom assemblages on macrophytes were significantly different from those on wood and rocks (ANOSIM R = 0.63, P < 0.01). A redundancy analysis (RDA) of the 41‐lake data set identified BiomEpiV as a significant (P < 0.05) variable in structuring sedimentary diatom assemblages. The MRT analysis classified the lakes into three groups. These groups were (A) high‐macrophyte, nutrient‐limited lakes (BiomEpiV ≥525 μg · L^?1; total phosphorus [TP] <35 μg · L^?1; 23 lakes); (B) low‐macrophyte, nutrient‐limited lakes (BiomEpiV <525 μg · L^?1; TP <35 μg · L^?1; 12 lakes); and (C) eutrophic lakes (TP ≥35 μg · L^?1; six lakes). A semiquantitative model correctly predicted the MRT group of the lake 71% of the time (P < 0.001). These results suggest that submerged macrophytes have a significant influence on diatom community structure and that sedimentary diatom assemblages can be used to infer past macrophyte abundance.     

Physiological and morphological adaptations of herbaceous perennial legumes allow differential access to sources of varyingly soluble phosphate

The aim of this study was to investigate the capacity of three perennial legume species to access sources of varyingly soluble phosphorus (P) and their associated morphological and physiological adaptations. Two Australian native legumes with pasture potential (Cullen australasicum and Kennedia prostrata) and Medicago sativa cv. SARDI 10 were grown in sand under two P levels (6 and 40 µg P g^?1) supplied as Ca(H₂PO₄)₂·H₂O (Ca‐P, highly soluble, used in many fertilizers) or as one of three sparingly soluble forms: Ca₁₀(OH)₂(PO₄)₆ (apatite‐P, found in relatively young soils; major constituent of rock phosphate), C₆H₆O₂₄P₆Na₁₂ (inositol‐P, the most common form of organic P in soil) and FePO₄ (Fe‐P, a poorly‐available inorganic source of P). All species grew well with soluble P. When 6 µg P g^?1 was supplied as sparingly soluble P, plant dry weight (DW) and P uptake were very low for C. australasicum and M. sativa (0.1–0.4 g DW) with the exception of M. sativa supplied with apatite‐P (1.5 g). In contrast, K. prostrata grew well with inositol‐P (1.0 g) and Fe‐P (0.7 g), and even better with apatite‐P (1.7 g), similar to that with Ca‐P (1.9 g). Phosphorus uptake at 6 µg P g^?1 was highly correlated with total root length, total rhizosphere carboxylate content and total rhizosphere acid phosphatase (EC 3.1.3.2) activity. These findings provide strong indications that there are opportunities to utilize local Australian legumes in low P pasture systems to access sparingly soluble soil P and increase perennial legume productivity, diversity and sustainability.     

PHOSPHORUS AND NITROGEN NUTRITION IN CHONDRUS CRISPUS (RHODOPHYTA): EFFECTS ON TOTAL PHOSPHORUS AND NITROGEN CONTENT,CARRAGEENAN PRODUCTION,AND PHOTOSYNTHETIC PIGMENTS AND METABOLISM1

The existence of a phenomenon in phosphorus (P) nutrition comparable to the “Neish effect” in nitrogen (N) nutrition (an inverse relation between seawater N enrichment and carrageenan content) was investigated in the temperate red alga Chondrus crispus Stackhouse. Plants were preconditioned for 17 d and then cultured under varying enrichments of P (0, 3, 6, 10, 15 μM P·wk^?1) and a constant N enrichment (53.5 μM N·wk^?1) for 5 wk. Tissue total P, tissue total N, and carrageenan contents were then determined. Identical experiments were performed using C. crispus collected during the fall, winter, spring, and summer seasons. The procedure was repeated using material collected during the following fall season and cultured under constant P (6 μM P·wk^?1) and varying N enrichments (0, 3, 6, 10, 25 μM N·wk^?1). In the fall (P) experiment, carrageenan content was the highest [53.1 ± 0.3% DW (dry weight)], and tissue total P content was the lowest (1.71 ± 0.27 mg P·g DW^?1) in plants that received no P enrichment. Carrageenan content was stable (46.1 ± 1.8% DW) for plants given enrichments of 3 μM P·wk^?1 and greater. Thus, a decrease in carrageenan content, concomitant with an increase in tissue total P content, was observed, but only at tissue total P levels below 2 mg P·g DW^?1. As these levels were always higher than 2 mg P·g DW^?1 in the winter, spring, and summer experiments, carrageenan content remained constant within each season at 46.2 ± 1.3, 43.1 m 0.7, and 44.5 ± 0.6% DW, respectively. Nitrogen enrichment of plants collected in the fall did not affect carrageenan content, which was stable at 49.3 ± 0.9% DW. When these plants were compared with those of the previous fall experiment (6 μM P·wk^?1 and 53.5 μM N·wk^?1), a slight increase in carrageenan content was noted. Thus, at sufficiently high concentration, N also decreased carrageenan content in C. crispus. Phosphorus nutrition had no significant effect on photosynthesis versus irradiance parameters (P_max, α, R_d, I_c, and I_k), the contents of the photosynthetic pigments chlorophyll-a, phycoerythrin (PE), phycocyanin (PC), and allophycocyanin (APC), and the ratios PE:APC and PC:APC. In contrast, N nutrition affected both P_maxand the photosynthetic pigment contents. The data indicate that N limitation reduces the number of phycobilisomes but not their size. The greater reduction in phycobiliprotein than chlorophyll-acontent corroborates the natural bleaching phenomenon regularly observed in C. crispus populations during summer when N levels are generally low in seawater. These results suggest that C. crispus in the temperate waters of the Bay of Fundy may experience N limitation, but P limitation is unlikely.     

Positive effect of shredders on microbial biomass and decomposition in stream microcosms

1. Animals play a major role in nutrient cycling via excretory processes. Although the positive indirect effects of grazers on periphytic algae are well understood, little is known about top‐down effects on decomposers of shredders living on leaf litter. 2. Nutrient cycling by shredders in oligotrophic forest streams may be important for the microbial‐detritus compartment at very small spatial scales (i.e. within the leaf packs in which shredders feed). We hypothesised that insect excretion may cause local nutrient enrichment, so that microorganism growth on leaves is stimulated. 3. We first tested the effect of increasing concentration of ammonium (+10, +20 and +40 μg NH₄⁺ L^?1) on fungal and bacterial biomass on leaf litter in a laboratory experiment. Then we performed two experiments to test the effect of the presence and feeding activity of shredder larvae. We used two species belonging to the trichopteran family Sericostomatidae: the Palaearctic Sericostoma vittatum and the Neotropical Myothrichia murina, to test the effect of these shredders on fungal and bacterial biomass and decomposition on leaves of Quercus robur and Nothofagus pumilio, respectively. All experiments were run in water with low ammonium concentrations (2.4 ± 0.34 to 14.47 ± 0.95 μg NH₄⁺ L^?1). 4. After 5 days of incubation, NH₄ concentrations were reduced to near‐ambient streamwater concentrations in all treatments with leaves. Fungal biomass was positively affected by increased ammonium concentration. On the other hand, bacteria abundance was similar in all treatments, both in terms of abundance (bacteria cells mg^?1 leaf DW) and biomass. However, there was a tendency towards larger mean cell size in treatments with 20 μg NH₄ L^?1. 5. In the experiment with S. vittatum, fungal biomass in the treatment with insects was more than twice that in the control after 15 days. Bacteria were not detected in treatments with insects, where hyphae were abundant, but they were abundant in treatments without larvae. In the decomposition experiment run with M. murina, leaf‐mass loss was significantly higher in treatments with larvae than in controls. 6. Our hypothesis of a positive effect of shredders on fungal biomass and decomposition was demonstrated. Insect excretion caused ammonium concentration to increase in the microcosms, contributing to microbial N uptake in leaf substrata, which resulted in structural and functional changes in community attributes. The positive effect of detritivores on microbes has been mostly neglected in stream nutrient‐cycling models; our findings suggest that this phenomenon may be of greater importance than expected in stream nutrient budgets.     

The influence of light on copper‐limited growth of an oceanic diatom,Thalassiosira oceanica (Coscinodiscophyceae)

Thalassiosira oceanica (CCMP 1005) was grown over a range of copper concentrations at saturating and subsaturating irradiance to test the hypothesis that Cu and light were interacting essential resources. Growth was a hyperbolic function of irradiance in Cu‐replete medium (263 fmol Cu′ · L^?1) with maximum rates achieved at 200 μmol photons · m^?2 · s^?1. Lowering the Cu concentration at this irradiance to 30.8 fmol Cu′ · L^?1 decreased cellular Cu quota by 7‐fold and reduced growth rate by 50%. Copper‐deficient cells had significantly slower (P < 0.0001) rates of maximum, relative photosynthetic electron transport (rETR_max) than Cu‐sufficient cells, consistent with the role of Cu in photosynthesis in this diatom. In low‐Cu medium (30.8 fmol Cu′ · L^?1), growth rate was best described as a positive, linear function of irradiance and reached the maximum value measured in Cu‐replete cells when irradiance increased to 400 μmol photons · m^?2 · s^?1. Thus, at high light, low‐Cu concentration was no longer limiting to growth: Cu concentration and light interacted strongly to affect growth rate of T. oceanica (P < 0.0001). Relative ETR_max and Cu quota of cells grown at low Cu also increased at 400 μmol photons · m^?2 · s^?1 to levels measured in Cu‐replete cells. Steady‐state uptake rates of Cu‐deficient and sufficient cells were light‐dependent, suggesting that faster growth of T. oceanica under high light and low Cu was a result of light‐stimulated Cu uptake.     

Daphnia population growth but not moulting is a substantial phosphorus drain for phytoplankton

SUMMARY 1. Negative effects of zooplankton on the availability of phosphorus (P) for phytoplankton as a result of the retention of nutrients in zooplankton biomass and the sedimentation of exoskeletal remains after moulting, have been recently proposed. 2. In a mesocosm study, the relative importance of these mechanisms was tested for the freshwater cladoceran Daphnia hyalina×galeata. A total of 13 mesocosm bags was suspended in a mesotrophic German lake during summer 2000 and fertilised with inorganic P in order to obtain a total nitrogen to total P ratio closer to the Redfield ratio. D. hyalina×galeata was then added at a logarithmically scaled density gradient of up to 40 ind. L^?1. Zooplankton densities, dissolved inorganic, particulate organic (seston <100 μm), as well as total nutrient concentrations were monitored. Additionally, nutrient concentrations of sediment water removed from the bottom of the mesocosm bags via a manual pump were determined. 3. Seston carbon (C), seston P and total P were significantly negatively correlated with Daphnia densities. The amount of particulate P (～5–6 μg P L^?1) sequestered from the seston compartment by Daphnia corresponded roughly to the increase of zooplankton biomass (population growth). Soluble reactive phosphorous (SRP) was at all times high (～25–35 μg P L^?1) and possibly unavailable to phytoplankton as a result of P adsorption to calcite during a calcite precipitation event (whiting). P concentrations determined in sediment water were generally <60 μg P m^?2 and thus never exceeded 1% of the total amount of P bound in particulate matter of the overlying water column. 4. Seston C : P ratios followed a polynomial second‐order function: At Daphnia densities <40 ind. L^?1 a positive linear relationship was evident, which is explained by the stronger reduction of P compared with C in seston, and transfer of seston P to zooplankton. Highest seston C : P ratios of ～300 : 1 were observed at Daphnia densities of ～30–50 ind. L^?1, which is in agreement with proposed threshold values limiting Daphnia reproductive growth. At Daphnia densities >40–50 ind. L^?1 C : P ratios were decreased because of the strong reduction of seston C at close to constantly low seston P‐values of ～3–4 μg P L^?1. 5. At least for Daphnia, it may be concluded that – unlike population growth – the sedimentation of faecal pellets and carapaces after moulting seem negligible processes in pelagic phosphorus dynamics.     

SHIFT FROM CHLOROPHYTES TO CYANOBACTERIA IN BENTHIC MACROALGAE ALONG A GRADIENT OF NITRATE DEPLETION1

A survey of the spatial distribution of benthic macroalgae in a fluvial lake of the St. Lawrence River (Lake Saint‐Pierre, Quebec, Canada) revealed a shift in composition from chlorophytes to cyanobacteria along the flow path of nutrient‐rich waters originating from tributaries draining farmlands. The link between this shift and changes in water quality characteristics was investigated by sampling at 10 sites along a 15 km transect. Conductivity, current, light extinction, total phosphorus (TP; >25 μg P · L^?1), and ammonium (8–21 μg N · L^?1) remained fairly constant along the transect in contrast to nitrate concentrations, which fell sharply. Filamentous and colonial chlorophytes [Cladophora sp. and Hydrodictyon reticulatum (L.) Bory] dominated in the first 5 km where nitrate concentrations were >240 μg N · L^?1. A mixed assemblage of chlorophytes and cyanobacteria characterized a 1 km transition zone where nitrate decreased to 40–80 μg N · L^?1. In the last section of the transect, nitrate concentrations dropped below 10 μg N · L^?1, and cyanobacteria (benthic filamentous mats of Lyngbya wollei Farl. ex Gomont and epiphytic colonies of Gloeotrichia) dominated the benthic community. The predominance of nitrogen‐fixing, potentially toxic cyanobacteria likely resulted from excessive nutrient loads and may affect nutrient and trophic dynamics in the river.     

INORGANIC CARBON ACQUISITION BY CHRYSOPHYTES1

Twelve species, representing 12 families of the chrysophytes sensu lato, were tested for their ability to take up inorganic carbon. Using the pH‐drift technique, CO₂ compensation points generally varied between 1 and 20 μmol · L^?1 with a mean concentration of 5 μmol · L^?1. Neither pH nor alkalinity affected the CO₂ compensation point. The concentration of oxygen had a relatively minor effect on CO₂‐uptake kinetics, and the mean CO₂ compensation point calculated from the kinetic curves was 3.6 μmol · L^?1 at 10–15 kPa starting oxygen partial pressure and 3.8 μmol · L^?1 at atmospheric starting oxygen partial pressure (21 kPa). Similarly, uptake kinetics were not affected by alkalinity, and hence concentration of bicarbonate. Membrane inlet mass spectrometry (MIMS) in the presence and absence of acetazolamide suggested that external carbonic anhydrase in Dinobryon sertularia Ehrenb. and Synura petersenii Korschikov was either very low or absent. Rates of net HCO₃^? uptake were very low (～5% of oxygen evolution) using MIMS and decreased rather than increased with increasing HCO₃^? concentration, suggesting that it was not a real uptake. The CO₂ compensation points determined by MIMS for CO₂ uptake and oxygen evolution were similar to those determined in pH‐drift and were >1 μmol · L^?1. Overall, the results suggest that chrysophytes as a group lack a carbon‐concentrating mechanism (CCM), or an ability to make use of bicarbonate as an alternative source of inorganic carbon. The possible evolutionary and ecological consequences of this are briefly discussed.     

PRELIMINARY EVALUATION OF THE BIOREMEDIATION POTENTIAL OF PORPHYRA: PHOTOSYNTHETIC PRODUCTION BY BLADES AND CONCHOCELIS

Given their rapid growth and nutrient assimilation rates, Porphyra spp. are good candidates for bioremediation. The production potential of two northeast U.S. Porphyra species currently in culture (P. purpurea and P. umbilicalis) was evaluated by measuring rates of photosynthesis (as O₂ evolution) of samples grown at 20° C. Gametophytes of P. umbilicalis photosynthesized at rates that were 80% higher than those of P. purpurea over 5–20° C at both sub‐saturating and saturating irradiances (37 and 289 μmol photons m^?2 s^?1). Porphyra umbilicalis was both more efficient at low irradiances (higher alpha) and had a higher P_max than did P. purpurea (23.0 vs. 15.6 μmol O₂ g^?1 DW min^?1), suggesting that P. umbilicalis is a better choice for mass culture where self‐shading may be severe. The photosynthesis‐irradiance relationship for the Conchocelis stage of P. purpurea was also examined. Tufts of filaments, grown at 10, 15, and 20° C, were assayed at growth temperatures at irradiances ranging from 0–315 μmol photons m^?2 s^?1. Tufts were slightly more productive at 15° than at 10° C, but only ca. 4–6% as productive as gametophytes. Maximum rates of net photosynthesis were reduced by 66–74% in tufts grown at 20° C (only about 2% of gametophytes). The Conchocelis stage, however, need not limit mariculture operations; once Conchocelis cultures are established, they can be maintained over the long‐term as ready sources of spores for net seeding.     

Seasonal emission of monoterpenes by the Mediterranean tree Quercus ilex in field conditions: Relations with photosynthetic rates, temperature and volatility

The relationships of monoterpene emission with temperature, light, photosynthesis and stomatal conductance (g_s) were studied in Quercus ilex L. trees throughout the four annual seasons under field conditions. The highest monoterpene emission was measured in spring and summer (midday average of 11 μg [g DW]^?1 h^?1), whereas the lowest rates were found in autumn and winter (midday averages of 0.51 and 0.23 μg [g DW]^?1 h^?1, respectively). In spring and summer, limonene was the monoterpene emitted at highest rate (midday averages of 5.27–6.69 μg [g DW]^?1 h^?1), whereas α-pinene was emitted the most in autumn and winter (midday averages of 0.31 μg [g DW]^?1 h^?1). The monoterpenes limonene, α-pinene and β-pinene represented about 75–95% of total detected monoterpenes. The total monoterpene emission rates represented about 0.04% of carbon fixed in autumn, 0.17% in winter, 0.84–2.51% in spring and 1.22–5.13% in summer. Significant correlations of total monoterpene emission with temperature were found when considering either summer emission or the emission over the entire year, whereas significant correlations with net photosynthetic rates were only found when considering summer season. Among individual terpenes, the most volatile, α-pinene and β-pinene, were more correlated with temperature than with net photosynthetic rates whereas the less volatile limonene was more correlated with net photosynthetic rate. Thus, under field conditions it seems that dependency of monoterpene emission on photosynthetic rate or temperature is partly related with volatility of the compounds. Influences of seasonality, temperature, photosynthetic rates and volatility should be considered in inventories and models of emission rates in Mediterranean ecosystems.     

Simultaneous real‐time assay of copper and cadmium ions by infrared photo diode electrode implanted in the muscle of live fish

The electrical circuit of an infrared photodiode electrode (IPE) was used in the simultaneous assay of copper and cadmium ions. The electrode's cyclic voltammetry (CV), chronoamperometry and square‐wave (SW) stripping voltammetric optimum conditions were examined. Results for 0–160 mg L^?1 and 50–400 μg L^?1 SW Cu(II) Cd(II), the relative standard deviation of 0.158 Cu(II), 0.077 Cd(II) (n = 15) using 20.0 mg L^?1 have been obtained at optimum conditions. The low detection limit (S/N) was attained to be at 14.71 μg L^?1(2.31 × 10^?7 mol L^?1) Cu(II) and 18.42 μg L^?1(1.63 × 10^?7 mol L^?1) Cd(II). The handmade electrode was implanted deep in the muscle of live fish and interfaced with an electrochemical workstation. Real‐time analytical application was performed on the online assay of living tissue as the specimen was moving. The methods are deemed useful in interfaced assay for physiological control, nanodiode fabrication, and in the production of laboratory on a biochip. © 2009 Wiley Periodicals, Inc. J Biochem Mol Toxicol 23:256–262, 2009; Published online in Wiley InterScience ( www.interscience.wiley.com ). DOI 10.1002/jbt.20287     

ACCLIMATION OF NITRATE UPTAKE BY PHYTOPLANKTON TO HIGH SUBSTRATE LEVELS1

A literature review of data on nitrate uptake by phytoplankton suggests that nitrate levels above 20 μmol N·L^?1 generally stimulated uptake rates in cultured unicellular algae and natural phytoplankton communities. This phenomenon indicates that phytoplankton cells acclimate to elevated nitrate levels by increasing their uptake capacity in a range of concentrations previously considered to be saturating. Cyanobacteria and flagellates were found to present a considerable capacity for acclimation, with low (0.1–2 μmol N·L^?1) half‐saturation values (K_s) at low (5–20 μmol N·L^?1) substrate levels and high (1–80 μmol N·L^?1) K_s values at high (30–100 μmol N·L^?1) substrate levels. However, some diatom genera (Rhizosolenia, Skeletonema, Thalassiosira) also appeared to possess a low affinity nitrate uptake system (K_s between 18 and 120 μmol N·L^?1), which can help resolve the paradox of their presence in enriched seas. It follows that present models of nitrate uptake can severely underestimate the effects of high nitrate concentrations on phytoplankton dynamics and development. A more adequate approach would be to consider the possibility of multiphasic uptake involving several phase transitions as nitrate concentrations increased. Because it is a nonlinear phenomenon featuring strong thresholds, this effect appears to override that of other variables, such as irradiance, temperature, and cell size. Within the present context of eutrophication and for a range of concentrations that is becoming more and more ecologically relevant, equations are tentatively presented as a first approach to estimate K_s from ambient nitrate concentrations.     

Oxidative stress induces alkaloid production in Uncaria tomentosa root and cell cultures in bioreactors

The effect of oxidative stress on indole alkaloids accumulation by cell suspensions and root cultures of Uncaria tomentosa in bioreactors was investigated. Hydrogen peroxide (H₂O₂, 200 μM) added to U. tomentosa cell suspension cultures in shaken flasks induced the production of monoterpenoid oxindole alkaloids (MOA) up to 40.0 μg/L. In a stirred tank bioreactor, MOA were enhanced by exogenous H₂O₂ (200 μM) from no detection up to 59.3 μg/L. Root cultures grew linearly in shaken flasks with a μ=0.045 days^?1 and maximum biomass of 12.08±1.24 g DW/L (at day 30). Roots accumulated 3α‐dihydrocadambine (DHC) 2354.3±244.8 μg/g DW (at day 40) and MOA 348.2±32.1 μg/g DW (at day 18). Exogenous addition of H₂O₂ had a differential effect on DHC and MOA production in shaken flasks. At 200 μM H₂O₂, MOA were enhanced by 56% and DHC by 30%; while addition of 800 and 1000 μM H₂O₂, reduced by 30–40% DHC accumulation without change in MOA. Root cultures in the airlift reactor produced extracellular H₂O₂ with a characteristic biphasic profile after changing aeration. Maximum MOA was 9.06 mg/L at day 60 while at this time roots reached ca. 1 mg/L of DHC. Intracellular H₂O₂ in root cultures growing in the bioreactor was 0.87 μmol/g DW compared to 0.26 μmol/g DW of shaken flasks cultures. These results were in agreement with a higher activity of the antioxidant enzymes superoxide dismutase and peroxidase by 6‐ and 2‐times, respectively. U. tomentosa roots growing in the airlift bioreactor were exposed to an oxidative stress and their antioxidant system was active allowing them to produce oxindole alkaloids.     

The production and antiprotozoal activity of abietane diterpenes in Salvia austriaca hairy roots grown in shake flasks and bioreactor

The biomass and concentration of bioactive quinone methide-type diterpenes in hairy roots of Salvia austriaca were determined and compared with levels of these metabolites in roots of field-grown plants. The cultures were maintained in shake flasks and a nutrient sprinkle bioreactor. Diterpene production was more efficient in the shake flask root culture than the bioreactor one. Biomass and diterpene production within the shake flask culture was evaluated using Schenk and Hildebrandt (SH), Gamborg (B5), and woody plant medium (WPM), with both full- and half-strength macro and micronutrient concentrations (1/2 SH, 1/2 B5, and 1/2 WPM). Among the tested media, SH medium proved to be most effective for biomass and diterpene production. In this medium, the transformed roots accumulated the levels of taxodone (3.89?mg?g^?1 DW; equivalent to 63.3?mg?L^?1), taxodione (1.15?mg?g^?1 DW; equivalent to 17.4?mg?L^?1), 15-deoxy-fuerstione (2.15?mg?g^?1 DW; equivalent to 32.5?mg?L^?1), and 7-(2′-oxohexyl)-taxodione (0.076?mg?g^?1 DW; equivalent to 1.1?mg?L^?1). Three diterpenes were also detected in the roots of S. austriaca intact plants, but their concentrations were lower than those in hairy root culture. No 7-(2′-oxohexyl)-taxodione was found in the roots of field-grown plants. The hairy roots were able to maintain high metabolite levels even for 6 years of cultivation. Taxodone, taxodione, 15-deoxy-fuerstione, and 7-(2′-oxohexyl)-taxodione were tested for in vitro activity against Trypanosoma brucei rhodesiense, T. cruzi, and Plasmodium falciparum and their cytotoxicity was determined using L6 cells. Among these compounds, taxodione was the most active against T. brucei rhodesiense [IC₅₀?=?0.05?µM with high selectivity, selectivity index (SI)?=?38]. Taxodione was found to inhibit the growth of P. falciparum and T. cruzi by 50% at respective concentrations of 1.9 and 7.1?µM (SI values of 1.0 and 0.27). Other diterpenoids demonstrated weaker activity against tested parasites (IC₅₀ values ranging from 0.62 to 194.7?µM) and lower selectivity (SI value ranged from 0.4 to 5.0).     

EVALUATION OF COMPETITIVE ABILITY OF STAURASTRUM LUETKEMUELLERII (CHLOROPHYCEAE) AND MICROCYSTIS AERUGINOSA (CYANOPHYCEAE) UNDER P LIMITATION1

The desmid Staurastrum luetkemuellerii Donat et Ruttner and the cyanobacterium Microcystis aeruginosa Kütz. showed pronounced differences in chemical composition and ability to maintain P fluxes. The cellular P:C ratio (Q_p) and the surplus P:C ratio (Q_sp) were higher in M. aeruginosa, indicating a lower yield of biomass C per unit of P. The subsistence quota (Q_p) was 1.85 μg P·mg C^?1in S. luetkemuellerii and 6.09 μg P·mg C^?1in M. aeruginosa, whereas the respective Q_p of P saturnted organisms (Q_s) were 43 and 63 μg P·mg C^?1. These stores could support four divisions in S. luetkemuellerii and three divisions in M. aeruginosa, which suggests that the former exhibited highest storage capacity (Q_s/Q₀). M. aeruginosa showed a tenfold higher activity of alkaline phosphatase than S. luetkemuellerii when P starved. The optimum N:P ratio (by weight) was 5 in S. luetkemuellerii and 7 in M. aeruginosa. The initial uptake of P_i pulses in the organisms was not inhibited by rapid (<1 h) internal feedback mechanisms and the short term uptake rote could be expressed solely as a function of ambient P_i. The maximum cellular C-based uptake rate (V_m) in P starved M. aeruginosa was up to 50 times higher than that of S. luetkemuellerii. It decreased with increasing growth rate (P status) in the former species and remained fairly constant in the latter. The corresponding cellular P-based value (U_m= V_m/Q_p) decreased with growth rate in both species and was about 10 times higher in P started M. aeruginosa than in S. luetkemuellerii. The average half saturation constant for uptake (K_m) was equal for both species (22 μg P·L^?1) and varied with the P status. S. luetkemuellerii exhibited shifts in the uptake rate of P_i that were characterized by increased affinity (U^m/K^m) at low P_i, concentrations (<4 μg P·L^?1) compared to that at higher concentrations. The species thus was well adapted to uptake at low ambient P_i, but M. aeruginosa was superior in P_i uptake under steady state and transient conditions when the growth rate was lower than 0.75 d^?1. Moreover, M. aeruginosa was favored by pulsed addition of P_i. M. aeruginosa relpased P_i at a higher rate than S. luetkemuellerii. Leakage of P_i from the cells caused C-shaped μ vs. P_i curves. Therefore, no unique K_s for growth could be estimated. The maximum growth rate (μ_m) (23° C) was 0.94 d^?1for S. luetkemuellerii and 0.81 d^?1for M. aeruginosa. The steady state concentration of P_i (P*) was lower in M. aeruginosa than in S. luetkemuellerii at medium growth rates. The concentration of P_i at which the uptake and release of P_i was equal (P_c was, however, lower in S. luetkemuellerii.     

Further investigation of the roles of auxin and cytokinin in the NH4+-induced stimulation of nodulation using white clover transformed with the auxin-sensitive reporter GH3:gusA

Although mineral N (nitrate and ammonium) is believed to have generally negative effects on nodulation in legume–rhizobia symbioses, previous studies have shown that low, static concentrations of ammonium stimulate nodulation in pea, and that this enhancement may be due to an elevation in cytokinin to auxin levels in roots. Here, the effects of ammonium (0.0, 0.1, 0.5 and 2.5 mM) on nodulation and auxin levels were investigated in wild‐type (WT) white clover (Trifolium repens cv. Haifa) and its transformants (lines 38 and 41) which contain the auxin‐sensitive reporter gene (GH3:gusA). The effects of exogenous application (10^?10, 10^?9 and 10^?8 M) of the cytokinin 6‐benzylaminopurine (BAP) were also assessed. Whole‐plant nodulation (nodules plant^?1) and dry weight (DW)‐specific nodulation (nodules g^?1 root DW) were stimulated (up to 49%) in all white clover lines by 0.1 mM NH₄⁺. This represents the first confirmation of an NH₄⁺‐induced stimulation of DW‐specific nodulation in a species other than pea. At 2.5 mM NH₄⁺, the effect was lost on whole‐plant nodulation and was inhibitory on DW‐specific nodulation. Rhizobial inoculation resulted in a decline in the expression of GH3:gusA in root tips as expected; however, ammonium treatment did not affect GH3 expression in any root zones. Exogenous application of BAP at 10^?9 and 10^?8 M stimulated whole‐plant and DW‐specific nodulation in wild‐type white clover to a similar degree as treatment with 0.1 mM NH₄⁺. These results support our previous hypothesis that the stimulation of nodulation by low concentrations of ammonium involves the alteration of the ratio of cytokinin to auxin, specifically by increasing cytokinin.