Accumulation of cadmium by Dunaliella tertiolecta Batcher

Cultures of Dunaliella tertiolecta were exposed to five concentrationsof cadmium in solution (0.1, 0.5, 1, 5 and 10ppm(mg 1^-1)). Theaccumulation of cadmium by the algae was found to have two phases,an initial rapid uptake followed by a stabilisation of the cellularcadmium levels. D. tertiolecta concentrated cadmium from solution(cone, factor approx. 1350) at exposures up to 1 ppm Cd butexposure to the higher concentrations caused no further increasein the accumulated cadmium concentration of the algae whichreached a nmriimim at about 1.5 µg Cd mg¹ Dunaliella.     

Growth of Dunaliella tertiolecta and associated bacteria in photobioreactors

The aim of this study was to test three flat-plate photobioreactor configurations for cultivation of marine green alga Dunaliella tertiolecta under non-axenic growth conditions and to characterize and quantify the associated bacteria. The photobioreactor cultivations were conducted using tap water-based media. Static mixers intended to enhance mixing and light utilization did not generally increase algal growth at the low light intensities used. The maximum biomass concentration (measured as volatile suspended solids) and maximum specific growth rate achieved in the flat plate with no mixer were 2.9?g?l(-1) and 1.3?day(-1), respectively. Based on quantitative polymerase chain reaction, bacterial growth followed the growth of D. tertiolecta. Based on 16S rDNA amplification and denaturing gradient gel electrophoresis profiling, heterotrophic bacteria in the D. tertiolecta cultures mainly originated from the non-axenic algal inocula, and tap water heterotrophs were not enriched in high chloride media (3?% salinity). Bacterial communities were relatively stable and reproducible in all flat-plate cultivations and were dominated by Gammaproteobacteria, Flavobacteria, and Alphaproteobacteria.     

The effect of ionic stress on photosynthesis in Dunaliella tertiolecta

A comparison of the effects of ionic stress and an uncoupler on long-term fluorescence transients (the Kautsky effect) in the green alga Dunaliella tertiolecta indicated that the large quenching induced by ionic stress was caused by a pH gradient across the thylakoid membrane. This possiblity was given support by the increase in the slow phase of 3-(3,4-dichlorophenyl)-1,1-dimethylurea-induced fluorescence relaxation in algae subjected to ionic stress. Low-temperature fluorescence emission spectra indicated that salt stress enhanced photosystem-I emission in the dark, and a comparison of simultaneous emissions at 695 and 720 nm at room temperature indicated a further increase in photosystem-I emission during the fluorescence transients. Taken together with the decrease in the fast phase of 3-(3,4-dichlorophenyl)-1,1-dimethylurea-induced fluorescence relaxation in stressed algae, our results indicate that ionic stress stimulates cyclic electron flow, and that non-cyclic flow is inhibited. The effect of sucrose-induced osmotic stress was similar to, but less marked than, the effects of NaCl and KCl; the effect of decreasing the external salinity was small.Abbreviations DCMU 3-(3,4-dichlorophenyl)-1,1-dimethylurea - FCCP carbonylcyanide p-trifluoromethoxyphenylhydrazone - PSI, II photosystem I, II     

Control of misses in oxygen evolution by the oxido-reduction state of plastoquinone in Dunaliella tertiolecta

The way misses happen in oxygen evolution is subject to debate (Govindjee et al. 1985). We recently observed a linear lowering of the miss probability with the flash number (Meunier and Popovic 1989). Therefore, we investigated in Dunaliella tertiolecta the link between the average miss probability and the redox state of plastoquinone after n flashes. The effect of flashes was to oxidize the plastoquinone pool; we found that the oxidation of plastoquinone highly correlated (linear regression: R ²=0.996) with the lowering of the miss probability. The flash frequency was found to affect both the miss probability and the redox state of plastoquinone. When pre-flashes were given using a high flash frequency (10 Hz), the plastoquinone pool was oxidized and misses were low; however, if long dark intervals between flashes were used, the oxidizing effect of flashes was lost and the misses were high. We could not explain our results by assuming equal misses over all S-states; but unequal misses, caused by deactivations, were coherent with our results. We deduced that chlororespiration was responsible for the reduction of plastoquinone in the dark interval between flashes. We compared oxygen evolution with and without benzoquinone, using a low flash frequency (0.5 Hz) for maximum misses. Benzoquinone lowered the misses from 34% to 3%, and raised the amplitude of oxygen evolution by more than a factor of two (2). From this we deduced that the charge carrier C postulated to explain misses (Lavorel and Maison-Peteri 1983) did not account for more than 3% of miss probability in Dunaliella tertiolecta. These results indicate that the misses in oxygen evolution are controlled by the redox state of plastoquinone, through deactivations.     

Fatty acids and global metabolites profiling of Dunaliella tertiolecta by shifting culture conditions to nitrate deficiency and high light at different growth phases

In this study, Dunaliella tertiolecta cells were shifted to nitrate-deficient (ND) and/or high light (HL) conditions at different growth phases. Profiling of global metabolites and fatty acids of D. tertiolecta cells were performed by shifting cultivation condition at exponential (EX) and stationary (ST) phases. ND and/or HL induced saturation of fatty acids by increasing saturated fatty acid and monounsaturated fatty acid and decreasing polyunsaturated fatty acid in D. tertiolecta cultures. The relative levels of metabolites like proline, valine, isoleucine, and myoinositol, which act as osmolytes, and ergosterol and glycerol, which are related to the osmoregulatory mechanism, decreased to a greater extent by ND than HL shifts. In addition, the effects of these shifts at the EX phase were observed to be more significant in most of the fatty acids. The results of this research might be applicable to the interpretation of metabolic synthesis and fatty acid production, and to the optimization of cultivation conditions for the use of D. tertiolecta and other microalgae as biofuels or nutraceutical sources.     

Modelling the uptake of nitrate by a growing plant with an adjustable root nitrate uptake capacity

A new model is presented to predict the plant uptake of nitrate supplied by diffusion and mass flow to its roots. Plant growth, root-shoot ratio and the plant's nitrate uptake capacity are all set dependent on the plant's N nutrition state. By thoroughly integrating processes occurring in both plant and soil, the model enables to control the relative importance of both under a wide range of different nutritional scenarios.Soil parameters D₀ diffusion coefficient in water (m² day^-1) - D_e diffusion coefficient in soil (m² day^-1) - C nitrate concentration in soil (mol m^-3) - f tortuosity (-) - volumetric moisture content (-) - R radial distance from root axis (m) Plant parameters b₁, b₂ parameters of biomass partitioning Equation (10) - IR interroot distance (m) - K_mU Michaelis-Menten constant of the uptake system (mol m^-3) - K_mNRA Michaelis-Menten constant of nitrogen reduction system (mol g^-1) - k₁, k₂, k₃ parameters of growth model Equation (9) - L_v Root length density (m m^-3) - NO_{3 set} ^- Set point of the cytoplasmatic nitrate pool (mol g^-1 dw) - NO_{3 c} ^- cytoplasmatic nitrate concentration (mol g^-1 dw) - NO_{3 v} ^- vacuolar nitrate concentration (mol g^-1 dw) - NRA_max maximum nitrate reductase activity (mol g^-1 dw day^-1) - N_re reduced nitrogen content (mol) - N_remax maximum reduced N concentration in the plant (mol g^-1 dw) - P partitioning coefficient of nitrate between cyplasm and vacuole - R(1) root radius (m) - RGR relative growth rate (day^-1) - U uptake rate (mol day^-1 m^-2) - U_max maximum uptake rate (Eq. 6) (day^-1 m^-2) - Vo water flux at root surface (m day^-1) - W_r root dry weight (g) - W_sh shoot dry weight (g) - X model parameter: number of root compartments - Y model parameter: number of nodes     

Efficiency of N uptake by wheat,as affected by time and rate of application,using N15-labelled ammonium sulphate and sodium nitrate

Summary The efficiency of N uptake by wheat was studied in field experiment, using N¹⁵-labelled (NH₄)₂SO₄ and NaNO₃. The N rate used was 120 kg N/Ha. It was applied in three split doses (at seeding, tillering and boot stage), in two split doses (at seeding and tillering stage) and in a single dose at seeding.The N applied at seeding, tillering and boot stage was actively used by wheat. However N applied at tillering stage was utilized most effectively. The single application of N was least productive for grain yield. Splitting the N fertilization increased the efficiency of applied N. The NO₃–N was most productive when applied in three split applications and produced significantly higher yield than single application. The NH₄–N was not as effective and its splitting beyond two doses did not benefit wheat. The A values and calculated effective rate of application (E.R.A.) also established the superiority of NO₃–N over NH₄–N. The E.R.A. proved useful in evaluating time of application as well.     

Regulation of phytochelatin synthesis by zinc and cadmium in marine green alga,Dunaliella tertiolecta

Although Cd(2+) is a more effective inducer of phytochelatin (PC) synthesis than Zn(2+) in higher plants, we have observed greater induction of PC synthesis by Zn(2+) than Cd(2+) in the marine green alga, Dunaliella tertiolecta. To elucidate this unique regulation of PC synthesis by Zn(2+), we investigated the effects of Zn(2+) and Cd(2+) on the activities of both phytochelatin synthase (PC synthase) and enzymes in the GSH biosynthetic pathway. PC synthase was more strongly activated by Cd(2+) than by Zn(2+), but the difference was not very big. On the other hand, gamma-glutamylcysteine synthetase (gamma-ECS) and glutathione synthetase (GS) were activated by both heavy metals, but their activities were higher in Zn-treated cells than in Cd-treated cells. Dose-dependent stimulation of intracellular reactive oxygen species (ROS) production was observed with Zn(2+), but not Cd(2+) treatment. These results suggest that Zn(2+) strongly promotes the synthesis of GSH through indirect activation of gamma-ECS and GS by stimulating ROS generation. This acceleration of the flux rate for GSH synthesis might mainly contribute to high level PC synthesis.     

The Growth and Intracellular Ionic Composition of Dunaliella tertiolecta in Magnesium-Rich Media

Dunaliella tertiolecta grew in a medium that contained MgSO₄(MgSO₄ medium), while this alga did not grow at all in mediathat contained MgCl₂ or Mg(NO₃)₂. The growth in the MgSO₄mediumwas inhibited in comparison with that in media that containedsodium salts, such as NaCl, NaNO₃, and Na₂SO₄. The energy chargeobtained from measurements of levels of adenine nucleotidesby HPLC were almost constant in Na- and Mg-containing media(about 0.87), indicating that the failure of growth in MgCl₂medium and Mg(NO₃)₂ medium was not directly related to.changesin the energy metabolism. K⁺ and Mg²⁺ were the dominant intracellularcations not only in Na-containing media (Na-media) but alsoin Mg-cohtaining media (Mg-media). The intracellular concentrationof Ca²⁺ was lower in Mg-media (1.6 mM) than that in Na-media(6mM). The concentrations of HPO₄^2– in cells incubatedin Mg-media were lower (less than 60 mM) than those in Na-media(greater than 110 mM). By contrast, the intracellular concentrationof SO₄^2– was higher in a MgSO₄ medium (26 mM) than thatin a Na₂SO₄ medium (4 mM) which, at least, compensated by 40%for the decrease in HPO₄^2–. The ability to grow in a MgSO₄medium may be related to the high intracellular concentrationof SO₄^2–. (Received September 20, 1990; Accepted March 22, 1991)     

Enhancement of tolerance to heavy metals and oxidative stress in Dunaliella tertiolecta by Zn-induced phytochelatin synthesis

The synthesis of phytochelatins (PCs) in a marine alga, Dunalliela tertiolecta, is strongly induced by Zn. Pretreatment of the cells with Zn enhances the tolerance toward toxic heavy metals such as Cd, Hg, Cu, Pb, and arsenate. Moreover, the pretreatment also increases the tolerance toward oxidative stress caused by hydrogen peroxide or paraquat. In vitro analysis shows that PC is a stronger scavenger of hydrogen peroxide and superoxide radical than glutathione. These results suggest that PCs inducibly synthesized by Zn treatment could play a role not only in detoxification of heavy metals but also in mitigation of oxidative stress.     

Carbon dioxide utilisation of Dunaliella tertiolecta for carbon bio-mitigation in a semicontinuous photobioreactor

Bio-fixation of carbon dioxide (CO₂) by microalgae has been recognised as an attractive approach to offset anthropogenic emissions. Biological carbon mitigation is the process whereby autotrophic organisms, such as microalgae, convert CO₂ into organic carbon and O₂ through photosynthesis; this process through respiration produces biomass. In this study Dunaliella tertiolecta was cultivated in a semicontinuous culture to investigate the carbon mitigation rate of the system. The algae were produced in 1.2-L Roux bottles with a working volume of 1 L while semicontinuous production commenced on day 4 of cultivation when the carbon mitigation rate was found to be at a maximum for D. tertiolecta. The reduction in CO₂ between input and output gases was monitored to predict carbon fixation rates while biomass production and microalgal carbon content are used to calculate the actual carbon mitigation potential of D. tertiolecta. A renewal rate of 45 % of flask volume was utilised to maintain the culture in exponential growth with an average daily productivity of 0.07 g L^?1 day^?1. The results showed that 0.74 g L^?1 of biomass could be achieved after 7 days of semicontinuous production while a total carbon mitigation of 0.37 g L^?1 was achieved. This represented an increase of 0.18 g L^?1 in carbon mitigation rate compared to batch production of D. tertiolecta over the same cultivation period.     

Glycerol-3-phosphatase and the control of glycerol metabolism in Dunaliella tertiolecta cells

Glycerol-3-phosphatase (EC 3.1.3.2.1) was studied by following the release of radioactive glycerol from L-(U-¹⁴C)glycerol-3-phosphate in Dunaliella tertiolecta enzyme extracts. The reaction showed a neutral pH optimum and had an absolute requirement for Mg²⁺. The substrate saturation curve was hyperbolic with an apparent K _m value for glycerol-3-phosphate of 0.7 mM in the absence of phosphate. Inorganic orthophosphate was a competitive inhibitor of the enzyme with an estimated K _j of 0.1 mM. The glycerol-3-phosphatase reaction was blocked nearly completely by millimolar Ca²⁺ concentrations. Ca²⁺ inhibition did not depend on the presence of calmodulin in the reaction medium. The characteristics of glycerol-3-phosphatase are discussed in relation to the regulation of the cyclic glycerol metabolism in Dunaliella cells during periods of osmotic stress.     

Plant response to nitrate starvation is determined by N storage capacity matched by nitrate uptake capacity in two Arabidopsis genotypes

In a low-input agricultural context, plants facing temporalnutrient deficiencies need to be efficient. By comparing theeffects of NO-starvation in two lines of Arabidopsis thaliana (RIL282 and 432 from the Bay-0xShahdarapopulation), this study aimed to screen the physiological mechanismsallowing one genotype to withstand NO-deprivation better than another and to rate the relative importance of processessuch as nitrate uptake, storage, and recycling. These two lines,chosen because of their contrasted shoot N contents for identicalshoot biomass under N-replete conditions, underwent a 10 d nitratestarvation after 28 d of culture at 5 mM NO. It was demonstrated that line 432 coped betterwith NO-starvation, producing higher shoot and root biomass and sustaining maximal growthfor a longer time. However, both lines exhibited similar featuresunder NO-starvation conditions. In particular, the nitrate pool underwent the same drastic andearly depletion, whereas the protein pool was increased to asimilar extent. Nitrate remobilization rate was identical too.It was proportional to nitrate content in both shoots and roots,but it was higher in roots. One difference emerged: line 432had a higher nitrate content at the beginning of the starvationphase. This suggests that to overcome NO-starvation, line 432 did not directly rely on theN pool composition, nor on nitrate remobilization efficiency,but on higher nitrate storage capacities prior to NO-starvation. Moreover, the higher resistanceof 432 corresponded to a higher nitrate uptake capacity anda 2–9-fold higher expression of AtNRT1.1, AtNRT2.1, andAtNRT2.4 genes, suggesting that the corresponding nitrate transportersmay be preferentially involved under fluctuating N supply conditions. Key words: Arabidopsis thaliana, genetic variability, N partitioning, N recycling, N use efficiency, nitrate deficiency, nitrate remobilization rate, nitrate transporter gene expression, nitrogen reserves, plant development Received 12 July 2007; Revised 21 November 2007 Accepted 17 December 2007