Higher CO2 Fixation Rate Research Articles

Pilot-scale bioremediation of rare earths wastewater by Chlamydomonas sp. YC

The extraction of rare earth elements (REEs) through in-situ leaching with ammonium sulphate [(NH4)2SO4] had resulted in the production of a large volume of ammonium-rich wastewater, causing severe environmental pollution. This study aimed to assess the ability of an indigenous microalga Chlamydomonas sp. YC, isolated from REEs wastewater, to directly treat real REEs wastewater under outdoor conditions in 50 L airlift photobioreactors (AL-PBRs) and 5.0 m3 open race-way photobioreactors (ORWPs). Additionally, the harvested Chlamydomonas sp. YC biomasses from these two pilot photobioreactors were comprehensively analyzed to evaluate the nutritional values. The results showed that Chlamydomonas sp. YC in AL-PBRs exhibited higher biomass production (1.1 g/L), greater removal efficiencies in NH4+-N (24.9%) and total nitrogen (20.4%), as well as higher CO2 fixation rate (125.0 mg/(L·d)), compared to those of ORWPs. Moreover, the Chlamydomonas sp. YC biomasses obtained from the two pilot photobioreactors contained 44.5% and 49.4% protein, 9.1% and 14.3% lipids. Moreover, Chlamydomonas sp. YC in the two pilot photobioreactors displayed essential amino acid indexes (EAAI) of 0.900, which was higher than that of soybean protein (0.657), indicating superior nutritional values. In conclusion, the implementation of the process involving Chlamydomonas sp. YC in AL-PBRs under outdoor conditions holds promise as a coupled microalgal biotechnology for the simultaneous removal of NH4+-N from REEs wastewater, and the capture of CO2 for the production of valuable biomass.

Alternative electron pathways of photosynthesis power green algal CO2 capture.

Microalgae contribute to about half of global net photosynthesis, which converts sunlight into the chemical energy (ATP and NADPH) used to transform CO2 into biomass. Alternative electron pathways of photosynthesis have been proposed to generate additional ATP that is required to sustain CO2 fixation. However, the relative importance of each alternative pathway remains elusive. Here, we dissect and quantify the contribution of cyclic, pseudo-cyclic, and chloroplast-to-mitochondrion electron flows for their ability to sustain net photosynthesis in the microalga Chlamydomonas reinhardtii. We show that (i) each alternative pathway can provide sufficient additional energy to sustain high CO2 fixation rates, (ii) the alternative pathways exhibit cross-compensation, and (iii) the activity of at least one of the three alternative pathways is necessary to sustain photosynthesis. We further show that all pathways have very different efficiencies at energizing CO2 fixation, with the chloroplast-mitochondrion interaction being the most efficient. Overall, our data lay bioenergetic foundations for biotechnological strategies to improve CO2 capture and fixation.

High-efficient CO2-to-protein bioconversion by oleaginous Coccomyxa subellipsoidea using light quality shift and nitrogen supplementation strategy

Oleaginous microalgae are usually used in CO2-to-lipid bioprocess for biofuels production but still faced the low productivity of biomass and lipid with uncompetitive cost. A novel CO2-to-protein bioconversion pattern by oleaginous Coccomyxa subellipsoidea was implemented in the present work as high-value application of CO2. Results demonstrated that the highest CO2 fixation rate (RCO2, 0.74 g/L/d) and protein productivity (178 mg/L/d) with lower protein content (42.33% DW) were achieved under red light; but under blue light, the highest protein content (52.07% DW) were reached with lower productivity (156.94 mg/L/d) and RCO2 (0.52 g/L/d). The analysis of key genes transcription and enzymes activity revealed that red light boosted the photosynthetic activity and photoprotection contributing to the high-efficient CO2 fixation. Based on this, a tailored light quality shift and nitrogen supplementation in a two-phase culture (red light in phase 1; red: blue-1:1 with nitrate addition in phase 2) was developed, reaching the maximal biomass yield (6.12 g/L), RCO2 (0.90 g/L/d), protein content (52.00% DW) with protein productivity (265.83 mg/L/d) simultaneously, 2.0-fold, 1.2-fold, 2.9-fold and 2.3-fold higher than that under white light as the control. The highest contents of AAs (47.12% DW) and EAAs (18.67% DW) with essential amino acid index (1.3) were achieved, superior to FAO/WHO reference. Time-dependent gene transcription analysis revealed the up-regulated photosynthesis and nitrogen assimilation in the two-phase culture. This work demonstrated that the oleaginous C. subellipsoidea could be well used for protein production and CO2 fixation, providing a promising application approach in carbon neutrality and high-yield protein production.

Metabolic versatility enables sulfur-oxidizers to dominate primary production in groundwater

High rates of CO2 fixation and the genetic potential of various groundwater microbes for autotrophic activity have shown that primary production is an important source of organic C in groundwater ecosystems. However, the contribution of specific chemolithoautotrophic groups such as S-oxidizing bacteria (SOB) to groundwater primary production and their adaptation strategies remain largely unknown. Here, we stimulated anoxic groundwater microcosms with reduced S and sampled the microbial community after 1, 3 and 6 weeks. Genome-resolved metaproteomics was combined with 50at-% 13CO2 stable isotope probing to follow the C flux through the microbial food web and infer traits expressed by active SOB in the groundwater microcosms. Already after 7 days, 90% of the total microbial biomass C in the microcosms was replaced by CO2-derived C, increasing to 97% at the end of incubation. Stable Isotope Cluster Analysis revealed active autotrophs, characterized by a uniform 13C-incorporation of 45% in their peptides, to dominate the microbial community throughout incubation. Mixo- and heterotrophs, characterized by 10 to 40% 13C-incorporation, utilized the primarily produced organic C. Interestingly, obligate autotrophs affiliated with Sulfuricella and Sulfuritalea contained traits enabling the storage of elemental S in globules to maintain primary production under energy limitation. Others related to Sulfurimonas seemed to rapidly utilize substrates for fast proliferation, and most autotrophs further maximized their energy yield via efficient denitrification and the potential for H2 oxidation. Mixotrophic SOB, belonging to Curvibacter or Polaromonas, enhanced metabolic flexibility by using organic compounds to satisfy their C requirements. Time series data spanning eight years further revealed that key taxa of our microcosms composed up to 15% of the microbial groundwater community, demonstrating their in-situ importance. This showed that SOB, by using different metabolic strategies, are able to account for high rates of primary production in groundwater, especially at sites limited to geogenic nutrient sources. The widespread presence of SOB with traits such as S storage, H2 oxidation, and organic C utilization in many aquatic habitats further suggested that metabolic versatility governs S-fueled primary production in the environment.

Oleaginous Microalga Coccomyxa subellipsoidea as a Highly Effective Cell Factory for CO2 Fixation and High-Protein Biomass Production by Optimal Supply of Inorganic Carbon and Nitrogen.

Microalgae used for CO2 biofixation can effectively relieve CO2 emissions and produce high-value biomass to achieve “waste-to-treasure” bioconversion. However, the low CO2 fixation efficiency and the restricted application of biomass are currently bottlenecks, limiting the economic viability of CO2 biofixation by microalgae. To achieve high-efficient CO2 fixation and high-protein biomass production, the oleaginous microalga Coccomyxa subellipsoidea (C. subellipsoidea) was cultivated autotrophically through optimizing inorganic carbon and nitrogen supply. 0.42 g L−1 NaHCO3 supplemented with 2% CO2 as a hybrid carbon source resulted in high biomass concentration (3.89 g L−1) and productivity (318.33) with CO2 fixation rate 544.21 mg L−1 d−1 in shake flasks. Then, used in a 5-L photo-fermenter, the maximal protein content (60.93% DW) in batch 1, and the highest CO2 fixation rate (1043.95 mg L−1 d−1) with protein content (58.48% DW) in batch 2 of repeated fed-batch cultures were achieved under 2.5 g L−1 nitrate. The relative expression of key genes involved in photosynthesis, glycolysis, and protein synthesis showed significant upregulation. This study developed a promising approach for enhancing carbon allocation to protein synthesis in oleaginous microalga, facilitating the bioconversion of the fixed carbon into algal protein instead of oil in green manufacturing.

Evaluation of potent cyanobacteria species for UV-protecting compound synthesis using bicarbonate-based culture system.

The present investigation evaluates the potential of three cyanobacteria species Anabaena cylindrica, Nostoc commune and Synechococcus BDUSM-13 for photo-protecting mycosporine-like amino acids (MAAs) synthesis using bicarbonate-based culture system. Current investigations witnessed noteworthy bicarbonate tolerance of all species (NaHCO3; 0.5, 1 and 2gL- 1) in terms of their growth rate, chlorophyll content, biomass productivity and carbon fixation ability. Among all strains, Synechococcus BDUSM-13 showed maximum surge in specific growth rate (i.e. 0.72 day-1) at 1gL-1, productivity (i.e. 0.92 ± 0.06g day-1L-1) and chlorophyll content (i.e. 0.09gL-1) at 2g day-1L-1. Synechococcus cells were also has the 0.48gdw-1 carbon content with highest CO2 fixation rate (i.e. 0.653g.CO2mL-1day-1) at 2gL-1. Though, they were not able to produce MAAs after long UV-B exposure (i.e. 24 and 48h). A. cylindrica strain was the most competent species for the bicarbonate-based approach, produced UV-protecting iminomycosporine compound (i.e. shinorine, λ max at 334 ± 2nm) along with carbon fixation (i.e. 0.49gCO2mL-1day-1) at 2gL-1 NaHCO3. This suggests the bicarbonate supplementation during cultivation is a promising strategy to increase cellular abundance, biomass productivity and carbon fixation in cyanobacteria. However, UV-B irradiation may cause species-specific differences in the MAAs synthesis to produce UV-protecting compounds.

Nitrogen and 17β-Estradiol level regulate Thermosynechococcus sp. CL-1 carbon dioxide fixation, monosaccharide production, and estrogen degradation

Thermosynechococcus sp. CL-1 (TCL-1), a thermophilic cyanobacterium from a hot spring in Taiwan, has been known of its efficiency in CO2 fixation, byproducts production (pigments, macromolecules). This study observed the performance of TCL-1 in CO2 fixation, estrogen degradation, and monosaccharide production under various levels of Dissolved Inorganic Nitrogen (DIN) and 17β-estradiol (E2) as nitrogen supply and estrogen addition. Under nitrogen starvation, TCL-1 performed similar results on CO2 fixation rate and biomass production but enhanced the monosaccharide production compared to the cases of high nitrogen supply. The highest CO2 fixation rate and glucose productivity reached to 151.8 ± 6.6 and 38.1 ± 0.9 mg/L/h, under DIN level of 0.58 mM and 0.5 mg/L E2. Adding E2 in the system did not inhibit the performance of TCL-1. During the cultivation, TCL-1 converted E2 into E1 and the biodegradation was the main path for estrogen degradation. Total E2 degradation reached to 69.4 ± 2.0%.

Effects of CO2 concentration on carbon fixation capability and production of valuable substances by Spirulina in a columnar photobioreactor

Three cyanobacterial strains of the genus Spirulina, i.e., LAMB171, LAMB172, and LAMB220, were cultivated at different CO2 concentrations in a columnar photobioreactor to determine the most optimal strain for CO2 mitigation and co-production of bioactive substances, specifically carotenoids, C-phycocyanin and allophycocyanin (A-PC). The best biomass production and the highest CO2 fixation rate were obtained at 10% CO2 concentration for all three Spirulina strains evaluated in the present study. When compared with the control group, the contents of chlorophyll a, carotenoids, C-PC, and A-PC were more optimal in the investigated Spirulina strains at lower CO2 concentrations (2% and 5%). However, the content of these bioactive substances was not significantly different at a 10% CO2 concentration compared with that at lower CO2 concentrations. Overall, it was determined that 10% CO2 concentration was noted to be most suitable for the production of these bioactive substances owing to the highest biomass productivity for all three Spirulina strains. Besides, LAMB220 was found to be the most effectively mitigated CO2 in three strains on account of its high biomass productivity and excellent C-fixation capability, while LAMB171 and LAMB172 species were more suitable for the production of pigments at a 10% CO2 concentration.

Improved photosynthetic characteristics of Chlorella mutant MS700 induced by nuclear radiation

Chlorophyll a variable fluorescence and oxygen evolution were measured in microalgae Chlorella and its mutant MS700, which was mutated by nuclear radiation with an increased biomass yield. The mutant MS700 showed an improvement in photosynthetic characteristics under various CO2 concentrations. Results showed that chlorophyll-based flash oxygen yield was 104% higher in MS700 than in the wild type, indicating more available electron acceptors in the plastoquinone pool and a higher light to chemical energy conversion efficiency. The higher chlorophyll a variable fluorescence at the end of PQ pool-regulated phase implied a more robust mechanism for consumption of electrons generated in PSII, indicating a more efficient downstream process. Monitoring of CO2 fixation in the Calvin cycle showed that the mutant had a 27.7% higher capability of incorporating its dissolved inorganic carbon. The oxygen evolution rate was 31.4% higher in MS700, which also indicated a higher CO2 fixation rate. These results consistently demonstrated carbon fixation efficiency in the mutant MS700, resulting in the higher biomass yield.

Sulfurimonas subgroup GD17 cells accumulate polyphosphate under fluctuating redox conditions in the Baltic Sea: possible implications for their ecology.

The central Baltic Sea is characterized by a pelagic redox zone exhibiting high dark CO2 fixation rates below the chemocline. These rates are mainly driven by chemolithoautotrophic and denitrifying Sulfurimonas GD17 subgroup cells which are motile and fast-reacting r-strategists. Baltic Sea redox zones are unstable and a measurable overlap of nitrate and reduced sulfur, essential for chemosynthesis, is often only available on small scales and short times due to local mixing events. This raises the question of how GD17 cells gain access to electron donors or acceptors over longer term periods and under substrate deficiency. One possible answer is that GD17 cells store high-energy-containing polyphosphate during favorable nutrient conditions to survive periods of nutrient starvation. We used scanning electron microscopy with energy-dispersive X-ray spectroscopy to investigate potential substrate enrichments in single GD17 cells collected from Baltic Sea redox zones. More specific substrate enrichment features were identified in experiments using Sulfurimonas gotlandica GD1T, a GD17 representative. Sulfurimonas cells accumulated polyphosphate both in situ and in vitro. Combined genome and culture-dependent analyses suggest that polyphosphate serves as an energy reservoir to maintain cellular integrity at unfavorable substrate conditions. This redox-independent energy supply would be a precondition for sustaining the r-strategy lifestyle of GD17 and may represent a newly identified survival strategy for chemolithoautotrophic prokaryotes occupying eutrophic redox zones.

Two-stage mixotrophic cultivation for enhancing the biomass and lipid productivity of Chlorella vulgaris

This study proposes a two-stage mixotrophic process for cultivating Chlorella vulgaris. Heterotrophic growth is the dominant step in Phase I (to increase microalgal biomass) and photoautotrophic growth occurs in Phase II (to improve biomass concentration and lipid production). The results show that the addition of the low-cost antioxidant sodium erythorbate (8 g L−1) significantly accelerates the growth of microalgae in the first stage with air aeration. Furthermore, a higher CO2 fixation rate was obtained in the second stage (at least 344.32 mg CO2 L−1 day−1) with 10% CO2 aeration. This approximately corresponds to an increase of 177% over simple photoautotrophic cultivation with 10% CO2 aeration during the whole period. The two-stage cultivation strategy achieved a maximum C. vulgaris biomass concentration of 3.45 g L−1 and lipid productivity of 43.70 mg L−1 day−1, which are 1.85 and 1.64 times those arising due to simple photoautotrophy, respectively. Moreover, an analysis of the product’s fatty acid profile indicates that C. vulgaris might be an ideal candidate for two-stage mixotrophic cultivation of a renewable biomass for use in biodiesel applications. Another interesting point to note from the study is that it is an insufficiency of N and CO2 that probably limits the further growth of C. vulgaris.

Culture characteristics of the atmospheric and room temperature plasma-mutated Spirulina platensis mutants in CO2 aeration culture system for biomass production

For biomass production of Spirulina platensis as feedstock of fermentation, the culture characteristics of three typical mutants of 3-A10, 3-B2 and 4-B3 generated by atmospheric and room temperature plasmas (ARTP) mutagenesis were systematically studied by using CO2 aeration culture system and compared with the wild strain. The specific growth rate of wild strain in the pure air aeration culture system exhibited a 76.2% increase compared with static culture, while the specific growth rates of the 3-A10, 3-B2 and 4-B3 in pure air aeration culture system were increased by 114.4%, 95.9% and 88.2% compared with their static cultures. Compared with static culture, the carbohydrate contents of wild strain, 3-A10, 3-B2 and 4-B3 in pure air aeration culture system dropped plainly by 51.0%, 79.3%, 85.5% and 26.1%. Increase of CO2 concentration enhanced carbohydrate content and productivity. Based on the carbohydrate productivity, the optimal inlet of CO2 concentration in aeration culture was determined to be 12% (v/v). Under this condition, 3-B2 exhibited the highest carbohydrate content (30.7%), CO2 fixation rate (0.120gCO2·g(-1)·d(-1)) and higher growth rate (0.093gL(-1)·d(-1)), while 3-A10 showed the highest growth rate (0.118gL(-1)·d(-1)) and higher CO2 fixation rate (0.117gCO2·g(-1)·d(-1)) but low carbohydrate content (24.5%), and 4-B3 showed the highest chlorophyll (Chl) content (3.82mg·g(-1)). The most outstanding mutant by static culture in terms of growth rate and carbohydrate productivity (3-B2), was also demonstrated by CO2 aeration culture system. This study revealed that the ARTP mutagenesis could generate the S.platensis mutants suitable for CO2 aeration culture aiming at biomass production.

Regulation of carbon metabolic fluxes in response to CO2 supplementation in phototrophic Chlorella vulgaris: a cytomic and biochemical study

As one of the promising species of microalgae for biofuel production, Chlorella vulgaris CS-42 was cultivated phototrophically in two cylindrical photobioreactors with aeration of 5 % (v/v) CO2 or air for 13 days to evaluate the effects of CO2 supplementation on biomass, CO2 fixation performance, and biochemical content. Significant increases of specific growth rate and total carbon content in biomass resulting in a higher CO2 fixation rate were found with 5 % CO2. The maximum biomass concentration, carbohydrate and fatty acid contents with 5 % CO2 were significantly higher than those with air, while carbohydrate biosynthesis was most affected as compared to other biochemical components. Cytomic analysis revealed a rapid accumulation of neutral lipid in the late growth phase with more lipid bodies visualized by confocal laser scanning microscopy (CLSM), when nitrate consumption was accelerated with CO2 supplementation. Gas chromatography mass spectrometry (GC-MS) analysis indicated that 5 % CO2 favored the formation of C18:2, which led to a decrease in the degree of lipid unsaturation (DLU). These results proved that CO2 supplementation was one of the most efficient methods to significantly prompt the growth of microalgae and increase the C/N ratio in the medium, which in turn regulated the carbon metabolic flux to enhance neutral lipid and fatty acid production in C. vulgaris.

The rise of the photosynthetic rate when light intensity increases is delayed in ndh gene-defective tobacco at high but not at low CO2 concentrations

The 11 plastid ndh genes have hovered frequently on the edge of dispensability, being absent in the plastid DNA of many algae and certain higher plants. We have compared the photosynthetic activity of tobacco (Nicotiana tabacum, cv. Petit Havana) with five transgenic lines (ΔndhF, pr-ΔndhF, T181D, T181A, and ndhF FC) and found that photosynthetic performance is impaired in transgenic ndhF-defective tobacco plants at rapidly fluctuating light intensities and higher than ambient CO2 concentrations. In contrast to wild type and ndhF FC, which reach the maximum photosynthetic rate in less than 1 min when light intensity suddenly increases, ndh defective plants (ΔndhF and T181A) show up to a 5 min delay in reaching the maximum photosynthetic rate at CO2 concentrations higher than the ambient 360 ppm. Net photosynthesis was determined at different CO2 concentrations when sequences of 130, 870, 61, 870, and 130 μmol m-2 s-1 PAR sudden light changes were applied to leaves and photosynthetic efficiency and entropy production (Sg) were determined as indicators of photosynthesis performance. The two ndh-defective plants, ΔndhF and T181A, had lower photosynthetic efficiency and higher Sg than wt, ndhF FC and T181D tobacco plants, containing full functional ndh genes, at CO2 concentrations above 400 ppm. We propose that the Ndh complex improves cyclic electron transport by adjusting the redox level of transporters during the low light intensity stage. In ndhF-defective strains, the supply of electrons through the Ndh complex fails, transporters remain over-oxidized (specially at high CO2 concentrations) and the rate of cyclic electron transport is low, impairing the ATP level required to rapidly reach high CO2 fixation rates in the following high light phase. Hence, ndh genes could be dispensable at low but not at high atmospheric concentrations of CO2.

Dynamics of metabolic activities and gene expression in the Roseobacter clade bacterium Phaeobacter sp. strain MED193 during growth with thiosulfate.

Metagenomic analyses of surface seawater reveal that genes for sulfur oxidation are widespread in bacterioplankton communities. However, little is known about the metabolic processes used to exploit the energy potentially gained from inorganic sulfur oxidation in oxic seawater. We therefore studied the sox gene system containing Roseobacter clade isolate Phaeobacter sp. strain MED193 in acetate minimal medium with and without thiosulfate. The addition of thiosulfate enhanced the bacterial growth yields up to 40% in this strain. Concomitantly, soxB and soxY gene expression increased about 8-fold with thiosulfate and remained 11-fold higher than that in controls through stationary phase. At stationary phase, thiosulfate stimulated protein synthesis and anaplerotic CO2 fixation rates up to 5- and 35-fold, respectively. Several genes involved in anaplerotic CO2 fixation (i.e., pyruvate carboxylase, propionyl coenzyme A [CoA], and crotonyl-CoA carboxylase) were highly expressed during active growth, coinciding with high CO2 fixation rates. The high expression of key genes in the ethylmalonyl-CoA pathway suggests that this is an important pathway for the utilization of two-carbon compounds in Phaeobacter sp. MED193. Overall, our findings imply that Roseobacter clade bacteria carrying sox genes can use their lithotrophic potential to gain additional energy from sulfur oxidation for both increasing their growth capacity and improving their long-term survival.

Microalgae-mediated simultaneous treatment of toxic thiocyanate and production of biodiesel

In this work, a method for simultaneously degrading the toxic pollutant, thiocyanate, and producing microalgal lipids using mixed microbial communities was developed. Aerobic activated sludge was used as the seed culture and thiocyanate was used as the sole nitrogen source. Two cultivation methods were sequentially employed: a lithoautotrophic mode and a photoautotrophic mode. Thiocyanate hydrolysis and a nitrification was found to occur under the first (lithoautotrophic) condition, while the oxidized forms of nitrogen were assimilated by the photoautotrophic consortium and lipids were produced under the second condition. The final culture exhibited good settling efficiency (∼ 70% settling over 10 min), which can benefit downstream processing. The highest CO2 fixation rate and lipid productivity were observed with 2.5% and 5% CO2, respectively. The proposed integrated algal-bacterial system appears to be a feasible and even beneficial option for thiocyanate treatment and production of microbial lipids.

Engineering strategies for improving the CO2 fixation and carbohydrate productivity of Scenedesmus obliquus CNW-N used for bioethanol fermentation

Engineering strategies were applied to improve the cell growth, CO2 fixation ability, and carbohydrate productivity of a Scenedesmus obliquus CNW-N isolate. The resulting carbohydrate-rich microalgal biomass was subsequently utilized as feedstock for ethanol fermentation. The microalga was cultivated with 2.5% CO2 in a photobioreactor on different operation modes. Semi-batch operations with 50% replacement of culture medium resulted in the highest CO2 fixation rate (1546.7 mg L(-1) d(-1)), carbohydrate productivity (467.6 mg L(-1) d(-1)), and bioethanol yield (0.202 g/g biomass). This performance is better than most reported values in the literature. The microalgal biomass can accumulate nearly 50% carbohydrates, as glucose accounted for nearly 80% of the total carbohydrate content. This glucose-predominant carbohydrate composition of the microalga is well suited for fermentative bioethanol production. Therefore, using the proposed carbohydrate-rich microalgal biomass both as the carbon sink and as the feedstock provides a feasible alternative to current carbon-reduction and bioethanol-production strategies.

Growth and carbon assimilation limitations in Ricinus communis (Euphorbiaceae) under soil water stress conditions

Water availability may influence plant carbon gain and growth, with large impacts on plant yield. Ricinus communis (L.), a drought resistant species, is a crop with increasing economic importance in Brazil, due to its use in chemical industry and for the production of biofuels. Some of the mechanisms involved in this drought resistance were analyzed in this study by imposing progressive water stress to pot-grown plants under glasshouse conditions. Water withholding for 53 days decreased soil water gravimetric content and the leaf water potential. Plant growth was negatively and significantly reduced by increasing soil water deficits. With irrigation suspension, carbon assimilation and transpiration were reduced and remained mostly constant throughout the day. Analysis of A/Ci curves showed increased stomatal limitation, indicating that limitation imposed by stomatal closure is the main factor responsible for photosynthesis reduction. Carboxylation efficiency and electron transport rate were not affected by water stress up to 15 days after withholding water. Drought resistance of castor bean seems to be related to a pronounced, early growth response, an efficient stomatal control and the capacity to keep high net CO2 fixation rates under water stress conditions.

Trophic ecology of massive shrimp aggregations at a Mid‐Atlantic Ridge hydrothermal vent site

The source of nutrition of shrimp that form giant aggregations at Mid‐Atlantic Ridge hydrothermal vents was explored by a combination of different molecular techniques. These animals have been hypothesized either to graze on the free‐living, surface microbial community or to feed off epibiotic bacteria growing on their exoskeleton. Stable isotope compositions of potential food sources, consumer tissues, and gut contents suggest that the shrimp derive most of their carbon and nitrogen from the epibionts. However, probing of nucleic acids extracted from shrimp guts points to the existence of a gut microflora that is isotopically similar but genetically distinct from the epibionts. Carbon dioxide fixation experiments were carried out to compare the magnitude of potential primary productivity of the different nutritional sources of the shrimp. Relatively low and comparable rates were observed for both the free‐living community and the epibionts but activity associated with gut preparations was surprisingly high. These high CO2 fixation rates in the gut may be indicative of a nutritional relationship between the shrimp and their gut microflora. Overall, the data indicate a nutritional symbiosis of the shrimp with their epibionts and possibly also with a separate gut microflora.

The biological CO2 fixation and utilization project by rite (2) — Screening and breeding of microalgae with high capability in fixing CO2 —

In order to mitigate the potential problems associated with increasing atmospheric CO2 levels, RITE has started the biological CO2 fixation and utilization project in 1990 using photosynthetic microorganisms, such as microalgae and photosynthetic bacteria. Photosynthetic microorganisms utilize flue gas CO2 as a carbon source and solar energy as the energy source to produce biomass which can be used for useful materials. Extensive screening has been conducted to obtain photosynthetic microorganisms from nature with high capability in fixing CO2 and/or with the ability to produce useful materials. More than ten strains of microalgae with high capability in fixing CO2 and/or with the ability to produce useful materials have been obtained. Two green algal strains obtained by the screening, Chlorella sp. UK001 and Chlorococcum littorale, showed the high CO2 fixation rates exceeding the initial target value of 1gCO2/1/day at 24h illumination. Botryococcus braunii SI-30 was obtained as the alga producing hydrocarbons which can be used as recyclable fuel resource. This alga contains more than 15% of its dry weight as hydrocarbons and shows relatively high growth rate than other Botryococcus braunii strains. Efforts are now under way to increase their CO2 fixation rates by optimizing the culture conditions and to develop applications for the products.