Effects of salinity, light and temperature on growth rates of two species of Gracilaria （Rhodophyta）

Effects of temperature, salinity and light intensity on growth rates of Gracilaria lichenoides and G. tenuistipitata var. liui Zhang et Xia were tested. Eight to ten levels of each factor were first tested separately. The best growth rate was obtained under the conditions of 32°C, 30 and 240 μmol/(m²·s) for G. lichenoides, and 24°C, 20 and 200 μmol/(m²·s) for G. tenuistipitata, respectively. Then a uniform design was used to evaluate the optimal combinations of the three factors. The best conditions for the highest daily specific growth rates (% increase in wet weight) are determined to be 31.30°C, 32.10, and 287.23 μmol/(m²·s) for G. lichenoides (16.26%/d), and 25.38°C, 21.10, and 229.07 μmol/(m²·s) for G. tenuistipitata (14.83%/d), respectively. Supported by the 908 Special Program (908-02-04-07), the National Basic Research Program of China (973 Program, No. 2006CB400608), and K. C. Wong Magna Fund in Ningbo University     

Inhibition of photosynthesis in the microalga Chaetoceros curvisetus （Bacillariophyta） by macroalga Gracilaria lemaneiformis （Rhodophyta）

We investigated the effects of dried macroalga Gracilaria lemaneiform （Rhodophyta） on photosynthesis of the bloom-forming microalga Chaetoceros curvisetus. C. curvisetus was cultured with different amounts of dried G. lemaneiformis under controlled laboratory conditions. We measured the photosynthetic oxygen evolution rate and established the chlorophyll a fluorescence transient （OJIP） curve coupled with its specific parameters. We observed concentration-dependent and time-dependent relationships between dried G. lemaneiformis and inhibition of photosynthesis in C. curvisetus. Co-culture with dried G. lemaneiformis also resulted in a decrease in the light-saturated maximum photosynthetic oxygen evolution rate （P~ax） in C. curvisetus, and a decrease in the OJIP curve along with its specific parameters; the maximum photochemical efficiency of PSII （FJFm）, the amount of active PSII reaction centers per excited cross section at t=0 and t=--tFM （RC/CS0 and RC/CSm, respectively）, the absorption flux per excited cross section at t=0 （ABS/ CS0）, and the efficiency with which a trapped exciton moves an electron into the electron transport chain （~u0）. The dark respiration rate （Rd） increased in C. curvisetus co-cultured with dried G. lemaneiformis. The JIP-test and the oxygen evolution results indicated that dried G. lemaneiJbrmis decreased the number of active reaction centers, blocked the electron transport chain, and damaged the oxygen-evolving complex of C. curvisetus. This result indicated that dried fragments of G. lemaneiformis could effectively inhibit photosynthesis of C. curvisetus, and thus, could serve as a functional product to control and mitigate C. curvisetus blooms.     

Biomass and carbon storage of Gracilariopsis lemaneiformis （Rhodophyta） in Zhanshan Bay,Qingdao, China

Marine macroalgae can absorb carbon and play an important role in carbon sequestration. As an important economic macroalga, Gracilariopsis lemaneiformis has the potential to significantly affect carbon absorption and storage in wave-sheltered intertidal reef systems. However, detailed knowledge on seasonal biomass changes and carbon storage of G. lemaneiformis is lacking, especially in many small and scattered ecosystems. Considering the influence of human activities on wild distribution of G. lemaneiformis, the understanding of seasonal dynamics of an economically important species in nature is necessary. In this study, we first investigated seasonal variations in biomass, coverage area, and carbon storage during low tide from August 2011 to July 2012 in Zhanshan Bay, Qingdao, China. Furthermore, we estimated the carbon storage potential of wild G. lemaneiformis using light use efficiency(LUE). The results show that the standing biomass and coverage area changed significantly with season. However, seasonal variations in carbon content and water content were not obvious, with an average content of 35.1% and 83.64%, respectively. Moreover, carbon storage in individual months varied between 0.67 and 47.03 g C/m 2, and the value of carbon storage was the highest in August and June and the lowest in February. In Zhanshan Bay, LUE of G. lemaneiformis was only 0.23%. If it is increased to the theoretical maximum(5%–6%), the carbon storage will have an increase of at least 21 times compared with the current, which suggested that carbon storage of wild G. lemaneiformis had a high enhancement potential. The study will help to assess a potential role of G. lemaneiformis in reducing atmospheric CO2.     

Optimization of conditions for tetraspore release and assessment of photosynthetic activities for different generation branches of Gracilaria lemaneiformis Bory

Gracilaria lemaneiformis Bory is an economically important alga that is primarily used for agar production. Although tetraspores are ideal seeds for the cultivation of G. lemaneiformis, the most popular culture method is currently based on vegetative fragments, which is labor-intensive and time-consuming. In this study, we optimized the conditions for tetraspore release and evaluated the photosynthetic activities of different colonies formed from the branches of G. lemaneiformis using a PAM (pulse-amplitude-modulated) measuring system. The results showed that variations in temperature and salinityhad significant effects on tetraspore yield. However, variations in the photon flux density (from 15 μmol m⁻² s⁻¹ to 480 μmol m⁻² s⁻¹) had no apparent effect on tetraspore yield. Moreover, the PAM-parameters Y(I), Y(II), ETR(I), ETR(II) and F _v/F _m of colonies formed from different branches showed the same trend: parameter values of first generation branches>second generation branches>third generation branches. These results suggest that the photosynthetic activities of different colonies of branches changed with the same trend. Furthermore, photosynthesis in G. lemaneiformis was found to be involved in vegetative reproduction and tetraspore formation. Finally, the first generation branches grew slowly, but accumulated organic compounds to form large numbers of tetraspores. Taken together, these results showed that the first generation branches are ideal materials for the release of tetraspores.     

Purification and characterization of a new thermostable κ-carrageenase from the marine bacterium Pseudoalteromonas sp. QY203

A new extracellular κ-carrageenase, namely CgkP, 34.0 kDa in molecular weight, was purified from Pseudoalteromonas sp. QY203. CgkP showed relatively high activity at acidities ranging from pH6.0 to pH9.0 and temperatures ranging from 30℃ to 50℃ with the highest activity at 45℃ and pH7.2. Sodium chloride increased its activity markedly, and KCl increased its activity slightly. The divalent and trivalent metal ions including Cu2+ , Ni2+ , Zn2+ , Mn2+ , Al3+ and Fe3+ significantly inhibited its activity, while Mg2+ did not. CgkP remained 70% of original activity after being incubated at 40℃ for 48 h, and remained 80% of the activity after being incubated at 45℃ for 1 h. It exhibited endo-κ-carrageenase activity, mainly depolymerizing the κ-carrageenan into disaccharide and tetrasaccharide. CgkP was more thermostable than most of previously reported κ-carrageenases with a potential of being used in industry.     

Preference of the herbivorous marine teleost Siganus canaliculatus for different macroalgae

The decomposition of a large amount of unexploited macroalgal resource along the coast of China often results in heavy environmental pollution. In order to pave a way of using macroalgae as the dietary ingredient of rabbitfish Siganus canaliculatus, one of a few farmed herbivorous marine teleosts in China, its preference （feeding selectivity） for different macroalgae was determined in this study. Seven seaweed species abundantly inhabiting the coast of east Guangdong Province were exposed simultaneously to rab-bitfish juveniles in laboratory （multiple-choice feeding） with their content and absolute intake assayed. It was found that the most preferred algae were Ulva prolifera, Gracilaria lemaneiformis and Chaetomorpha linum, less preferred algae were U. pertusa and Porphyra haitanensis, and least preferred ones were Sargassum fusiforme and Corallina sessilis. Such an order did not change when one to four relatively preferred seaweeds were removed. The preferred seaweeds were richer in protein and soluble sugar thus higher in energy than the least preferred. In addition, this fish was found to favor filamentous and flat algae rather than calcified ones. Ac-cordingly, the richness of nutrients and morphological characteristics determined the preference of S. canaliculatus for tested macro-algae.     

Analysis of DNA Methylation of <Emphasis Type="Italic">Gracilariopsis lemaneiformis</Emphasis> Under Temperature Stress Using the Methylation Sensitive Amplification Polymorphism (MSAP) Technique

Gracilariopsis lemaneiformis is an economically important agarophyte, which contains high quality gel and shows a high growth rate. Wild population of G. lemaneiformis displayed resident divergence, though with a low genetic diversity as was revealed by amplified fragment length polymorphism (AFLP) and simple sequence repeat (SSR) analyses. In addition, different strains of G. lemaneiformis are diverse in morphology. The highly inconsistence between genetic background and physiological characteristics recommends strongly to the regulation at epigenetic level. In this study, the DNA methylation change in G. lemaneiformis among different generation branches and under different temperature stresses was assessed using methylation sensitive amplified polymorphism (MSAP) technique. It was shown that DNA methylation level among different generation branches was diverse. The full and total methylated DNA level was the lowest in the second generation branch and the highest in the third generation. The total methylation level was 61.11%, 60.88% and 64.12% at 15°C, 22°C and 26°C, respectively. Compared with the control group (22°C), the fully methylated and totally methylated ratios were increased in both experiment groups (15°C and 26°C). All of the cytosine methylation/demethylation transform (CMDT) was further analyzed. High temperature treatment could induce more CMDT than low temperature treatment did.     

Effects of heavy metals （Pb^2＋ and Cd^2＋） on the ultrastructure, growth and pigment contents of the unicellular cyanobacterium Synechocystis sp. PCC 6803

The unicellular cyanobacterium Synechocystis sp. PCC 6803, a model organism known for its unique combination of highly desirable molecular genetic, physiological and morphological characteristics, was employed in the present study. The species was cultured in BG11 liquid medium contained various initial concentrations of Pb²⁺ and Cd²⁺ (0, 0.5, 1, 2, 4, 6 and 8 mg/L). The experiment was conducted for six days and the metal induced alterations in the ultrastructure, growth and pigment contents were assessed. Alterations in the ultrastructure of the Synechocystis sp. PCC 6803 cells became evident with the increased (>4 mg/L Pb²⁺) metal concentration. The photosynthetic apparatus (thylakoid membranes) were found to be the worst affected. Deteriorated or completely destroyed thylakoid membranes have made large empty spaces in the cell interior. In addition, at the highest concentration (8 mg/L Pb²⁺), the polyphosphate granules became more prominent both in size and number. Despite the initial slight stimulations (0.2, 3.8 and 6.5% respectively at 0.5, 1 and 2 mg/L Pb²⁺), both metals inhibited the growth in a dose-dependent manner as incubation progressed. Pigment contents (chlorophyll α, β carotene and phycocyanin) were also decreased with increasing metal concentration. Cells exposed to 6 mg/L Pb²⁺, resulted in 36.56, 37.39 and 29.34% reductions of chlorophyll α, β carotene and phycocyanin respectively over the control. Corresponding reductions for the same Cd²⁺concentrations were 57.83, 48.94 and 56.90%. Lethal concentration (96 h LC₅₀) values (3.47 mg/L Cd²⁺ and 12.11 mg/L Pb²⁺) indicated that Synechocystis sp. PCC 6803 is more vulnerable to Cd²⁺ than Pb²⁺. Supported by the Chinese Scholarship Council     

Toxic effects of Zn2+, Cu2+, Cd2+ and NH3 on Chinese prawn

This study deals with the toxic effects of Zn²⁺, Cu²⁺, Cd²⁺ and NH₃ on Chinese prawn (Penaeus orientalis). The median lethal concentrations of the toxic matters on Chinese prawn were determined. Cd²⁺>Cu²⁺>Zn²⁺>NH₃-Nm is the toxic order of these chemicals to Chinese prawn. The toxic order of concentrated Zn²⁺ and Cd²⁺ in tissues and organs of Chinese prawn is in the order. gill>shell+appendage>viscera>muscle.     

Removal of K^＋, Na^＋, Ca^2＋, and M^g2＋ from saline-alkaline water using the microalga Scenedesmus obliquus

The capability ofScenedesmus obliquus to remove cations （K^＋, Na^＋, Ca^2＋, Mg^2＋） from saline- alkaline water was investigated at different salinities （0, 5, 10, 15, 20, 25） and carbonate alkalinities （0, 5, 10, 15, 20, 25, 30, 35 mmol/L）. K^＋, Na^＋, Ca^2＋, and Mg^2＋ in saline-alkaline water were efficiently removed by S. obliquus. The maximum removal of the cations （29.37 mg for K^＋, 185.85 mg for Na^＋, 23.07 mg for Ca^2＋, 66.14 mg for Mg^2＋） occurred at salinity 25. The maximum removal of K^＋ （2.28 mg）, Na＋ （6.62 mg）, Ca^2＋ （1.01 mg）, and Mg2＋ （0.62 mg） occurred at carbonate alkalinities of 25 mmol/L for K＊, 35 mmol/L for Na＋, 20 mmol/L for Ca2＋, and 25 mmol/L for Mf＋, respectively. Under a salinity stress, the concentration of Na＇ in S. obliquus increased significantly, while that of K~ decreased significantly. The concentrations of Ca^2＋ and Mg2＋ decreased as well. The ratios of K＋/Na~, Ca2＋/Na^＋, and Mg^2＋/Na^＋ were significantly lower in all salinity treatments than those of the control. Under alkaline stress, the concentrations of Nan and K＋ in S. obliquus decreased significantly and the ratios of K^＋/Na^＋, Ca2＋/Na^＋, and Mg^2＋/Na^＋ were significantly higher in all treatments than in the control. Moreover, the concentrations of Ca2＋ and Mg2＋ in S. obliquus at alkalinities of 5-10 mmol/L were significantly higher than those of the other treatments. The removal of Na＋ by S. obliquus mainly occurs through biosorption, and Mg^2＋ and Ca^2 ＋ were removed through both biosorption and bioaccumulation.     

Ecological studies ofGracilaria asiatica andGracilaria lemaneiformis in Zhanshan Bay,Qingdao

Natural populations ofG. asiatica Zhang & Xia andG. lemaneiformis (Bory) Weber van Bosse were studied during 1984 and 1986 in Zhanshan Bay, Qingdao (36°4'N, 120° 21'E). Rapid growth (length, weight) of these plants occurred between mid-May and late June (water temperatures, 15–20°C). The major epiphyte ofG. asiatica wasEnteromorpha linza, whilePunctaria latifolia was the major epiphyte ofG. lemaneiformis. Epiphytism declined throughout early summer, and epiphytes were rare after mid-July (1984); they did not reappear in late summer, although macrophyte growth declined abruptly after early July. Populations ofG. asiatica varied during late spring-early summer between adjacent sandy and rocky portions of the intertidal zone; plants at the sandy site were larger and epiphyte-free. Amphipod densities were low on both species ofGracilaria, but the most abundant species wereAmpithoe lacertosa, Caprella equilibra, C. krøyeri, C. scauraandPontogeneia rostrata. Additional information on general community structure is provided for theG. asiatica zone.     

Effects of heavy metal ions (Cu2+, Pb2+ and Cd2+) on DNA damage of the gills, hemocytes and hepatopancreas of marine crab, Charybdis japonica

There are rising concerns about the hazardous effects of heavy metals on the environment. In this study, comet assay and DNA alkaline unwinding assay were conducted on the tissues (gills, hepatopancreas, and hemocytes) of Charybdis japonica in order to illustrate genotoxicity of three heavy metal ions (Cu2+, Pb2+, and Cd2+) on the marine crabs C. japonica. The crabs were exposed to Cu2+ (10, 50, and 100 ?g.L?1), Pb2+ (50, 250, and 500 ?g L?1) and Cd2+ (5, 25, and 50 ?g L?1), and the tissues were sampled at days 0.5, 1, 3, 6, 9, and 15. DNA alkaline unwinding assay was used for testing the DNA single strand break in gills and hepatopancreas and comet assay was employed for testing the DNA damage in hemocytes. The results showed that the DNA damage (F-value) of gills in the crabs exposed to the three heavy metals was decreased gradually during the exposure periods and there was a dose-time response relationship in certain time, suggesting that the levels of DNA single strand break in all the experimental groups increased significantly compared to the controls. Changes of F-value in hepatopancreas of the crabs exposed to the three heavy metals were similar to those in gills except that the peak values were found in the 500 ?g L?1 Pb2+ treatment group at day 3 and the 50 ?g L?1 Cd2+ treatment group at day 9. The ranks of DNA damage in gills and hepatopancreas induced by the three heavy metal ions (50 ?g L?1, day 15) were Cd2+ >Pb2+ >Cu2+ and Pb2+ >Cu2+ >Cd2+. The levels of DNA damage in gills were higher than those in hepatopancreas in the same experimental group. It can be concluded that indices of DNA damage can be used as the potential biomarkers of heavy metal pollution in marine environment.     

Phylogenetic analysis of epibacterial communities on the surfaces of four red macroalgae

Macroalgal surfaces are prone to being attached by bacteria. Epibacterial community structures on marine macroalgae are host-specific but temporally and spatially variable. In this study, we investigated the structure of epibacterial communities on the surfaces of four red macroalgae, Gracilaria lemaneiformis, Gloiopeltis furcata, Mazzaella sp. and Porphyra yezoensis, by analyzing the sequences of 16 S r RNA gene libraries. Healthy individuals of all macroalgae species were collected in winter from a farm at Dalian, China. The results showed that the epibacterial communities were mainly dominated by α-Proteobacteria, γ-Proteobacteria and Bacteroidetes. Deinococcus-Thermus, Spirochaetes and ε-Proteobacteria were also found. The majority of cloned sequences shared the greatest similarity to those of culturable organisms. A large portion of sequences from the α-Proteobacteria homed in Roseobacter clade, i.e., genera Ahrensia, Roseovarius, Litoreibacter, Octadecabacter, Thaiassobacter and Sulfitobacter, while members of Bacteroidetes mainly belonged to family Flavobacteriaceae. The cloned sequences could be separated into 66 OTUs at 0.01 distance value, and rare common OTUs were found among libraries. At genus level, Pseudoalteromonas dominated Gr. lemaneiformis and Gl. furcata libraries, accounting for 72.2% and 47.3%, respectively. Sulfitobacter dominated P. yezoensis library, accounting for 35.4%. A previously undefined cluster within Deinococcus-Thermus dominated Mazzaella sp. library, accounting for 24.6% of the all. These results indicated that a broad range of bacteria inhabited the surfaces of these macroalgae.     

Variations of antioxidant enzyme activity and malondialdehyde content in nemertean Cephalothrix hongkongiensis after exposure to heavy metals

The antioxidant enzyme activity and malondialdehyde (MDA) content of Cephalothrix hongkongiensis were studied to assess variations in the biochemical/physiological parameters of nemerteans under heavy metal stress. Worms were exposed to copper, zinc and cadmium solutions at different concentrations, and the activity of three antioxidant enzymes, catalase (CAT), superoxide dismutase (SOD), and glutathione peroxidase (GPX), and MDA content were measured. The results show that the activity of each enzyme changed immediately after exposure to heavy metals. CAT was invariably inhibited throughout the experimental period, while the SOD activity was significantly elevated by exposure to Cu²⁺ for 48 h, but then decreased. SOD was inhibited by Zn²⁺during the first 12 h of exposure, but activated when exposed for longer periods. Under Cd²⁺ stress, SOD activity decreased within 72 h. GPX activity varied greatly, being significantly increased by both Cu²⁺ and Zn²⁺, but significantly inhibited by Cd²⁺ in the first 12–24 h after exposure. MDA content increased on Cu²⁺ exposure, but normally decreased on Zn²⁺ exposure. MDA content followed an increase-decrease-increase pattern under Cd²⁺ stress. In conclusion, the antioxidant system of this nemertean is sensitive to heavy metals, and its CAT activity may be a potential biomarker for monitoring heavy metal levels in the environment.     

Application of RAPD in genetic diversity study onGracilaria lemaneiformis III. Phase and sex related markers

Random amplification of polymorphic DNA (RAPD) was conducted by using 10 random primers (P-1 to P-10) inGracilaria lemaneiformis. Phase and sex specific bands were amplified by primers P-2, P-6, P-7 and P-8: for P-2 a 1.4 kb band was found in female gametophytes and tetrasporophytes, for P-6, a 0.6 kb band appeared in male gametophytes and tetrasporophytes; for P-7, a 0.76 kb band appeared in male gametophytes and tetrasporophytes; for P-7, a 0.72 kb band appeared in female gametophytes and tetrasporophytes; for P-8, a 0.73 kb band only appeared in male gametophytes.

     

Production of dimethylsulfide and acrylic acid from dimethylsulfoniopropionate during growth of three marine microalgae

We measured the concentrations of dimethylsulfide （DMS）, acrylic acid （AA）, and dimethylsulfoniopropionate （DMSP） during growth of three microalgae： Prorocentrum micans, Gephyrocapsa oceanica, and Platymonas subcordiformis. The DMSP, AA, and DMS concentrations in culture media varied significantly among algal growth stages, with the highest concentrations in the late stationary growth stage or the senescent stage. In the stationary growth stage, the average DMSP concentration per cell in P. micans （0.066 5 pmol/cell） was 1.3 times that in G. oceanica （0.049 5 pmol/ cell） and 20.2 times that in P. subcordiformis （0.003 29 pmol/cell）. The average concentrations of AA were 0.044 6, 0.026 9, and 0.003 05 pmol/cell in P. micans, G. oceanica, and P. subcordiformis, respectively, higher than the concentrations of DMS （0.272, 0.497, and 0.086 2 fmol/cell, respectively）. There were significant positive correlations between cell density and AA, DMSP, and DMS concentrations. The ratios of DMS/AA and AA/（DMSP＋AA） in the three algae differed significantly over the growth cycle. In all three microalgae, the DMS/AA ratios were less than 25% during the growth period, suggesting that the enzymatic cleavage pathway, which generates DMS, was not the main DMSP degradation pathway. The changes in the DMS/AA ratio indicated that there was a higher rate of enzymatic breakdown of DMSP in the early growth period and a lower rate during senescence. In all three microalgae, the AA/（DMSP＋AA） ratio （degradation ratio of DMSP） decreased during the exponential growth phase, and then increased. The variations in these ratios can approximately indicate the cleavage mechanism of DMSP at different stages of algal growth.     

Genome size of <Emphasis Type="Italic">Alexandrium catenella</Emphasis> and <Emphasis Type="Italic">Gracilariopsis lemaneiformis</Emphasis> estimated by flow cytometry

Flow cytometry(FCM) technique has been widely applied to estimating the genome size of various higher plants. However, there is few report about its application in algae. In this study, an optimized procedure of FCM was exploited to estimate the genome size of two eukaryotic algae. For analyzing Alexandrium catenella, an important red tide species, the whole cell instead of isolated nucleus was studied, and chicken erythrocytes were used as an internal reference. The genome size of A. catenella was estimated to be 56.48 ± 4.14 Gb(1C), approximately nineteen times larger than that of human genome. For analyzing Gracilariopsis lemaneiformis, an important economical red alga, the purified nucleus was employed, and Arabidopsis thaliana and Chondrus crispus were used as internal references, respectively. The genome size of Gp. lemaneiformis was 97.35 ± 2.58 Mb(1C) and 112.73 ± 14.00 Mb(1C), respectively, depending on the different internal references. The results of this research will promote the related studies on the genomics and evolution of these two species.     

STUDY ON THE ISOTHERMS OF THE INTERACTION BETWEEN SUSPENDED PARTICLES AND Cu (

The isotherms of the interaction between the suspended particles and Cu²⁺, and the effects of lysine and asparaginic acid on the isotherms in the Huanghe (Yellow) River were studied by applying the theory and method of interfacial stepwise ion/coordination particle exchange. We obtained a new stepped river isotherm, formed by two curves joined together with a “plateau” in the middle. The adsorption equilibrium constantsK ₁ andK ₂ were calculated by using the isothermal equation of surface stepwise ion exchange. Amino acid in small amount promotes exchange adsorption of the suspended particles with Cu²⁺. The degree of promotive action relates to the isoelectric point of amino acid. The promotive effect of lysine is bigger than that of asparaginic acid. Project 29361001 supported by NSFC.     

Cloning and analysis of phycoerythrin genes inGracilaria Lemaneiformis (Rhodophyceae)

Cloning and sequencing of the genes coding for the α- and β- subunit of phycoerythrin (PE) of a red alga—Gracilaria lemaneiformis (GL) are reported. Alignment of 1084 nucleotides sequenced with three known red algal PE genes,Rhodella violacea (RV),Polysiphonia boldii (PB) andAglaothamnion neglectum (AN), showed high level of conservation, and similarities of 77,6% (between GL and RV), 77.9% (GL and AN) and 79.0% (GL and PB). The similarities of amino acids were 84.8% (between GL and RV), 85.7% (GL and PB), and 80.6% (AN and GL), higher than those among nucleotides. Project 39670405 supported by NSFC and Japanese Science Promotion Society (JSPS).     

Effects of iron electrovalence and species on growth and astaxanthin production of Haematococcus pluvialis

To increase the cell concentration and the accumulation of astaxanthin in the cultivation of Haematococcus pluvialis, effects of different iron electrovalencies (Fe²⁺-EDTA and Fe³⁺-EDTA) and species (Fe-EDTA, Fe(OH)_x^32x and FeC₆H₅O₇) addition on cell growth and accumulation of astaxanthin were studied. Results show that different iron electrovalencies have various effects on cell growth and astaxanthin accumulation of H. pluvialis. Compared with Fe³⁺-EDTA, Fe²⁺-EDTA stimulate more effectively the formation of astaxanthin. The maximum astaxanthin content (30.70 mg/g biomass cell) was obtained under conditions of 18 μmol/L Fe²⁺-EDTA, despite the lower cell density (2.3×10⁵ cell/ml) in such condition. Fe³⁺-EDTA is more effective than Fe²⁺-EDTA in improving the cell growth. Especially, the maximal steady-state cell density, 2.9×10⁵ cell/ml was obtained at 18 μmol/L Fe³⁺-EDTA addition. On the other hand, all the various species of iron (EDTA-Fe, Fe(OH)_x^32x, FeC₆H₅O₇) are capable to improve the growth of the algae and astaxanthin production. Among the three iron species, FeC₆H₅O₇ performed the best. Under the condition of a higher concentration (36 μmol/L) of FeC₆H₅O₇, the cell density and astaxanthin production is 2 and 7 times higher than those of iron-limited group, respectively. The present study demonstrates that the effects of the stimulation with different iron species increased in the order of FeC₆H₅O₇, Fe(OH)_x^32x and EDTA-Fe. Supported by Xiamen Scientific and Technologic Projects (No. 3052Z20031086), Xiamen University Alumni Association Foundation in Singapore, and the First National College Students Innovative Experimental Project