Partitioning of CO2 Fixation in the Colonial Cyanobacterium Microcystis aeruginosa: Mechanism Promoting Formation of Surface Scums

Constraints on inorganic carbon (C_i) availability stimulated buoyancy in natural, photosynthetically active populations of the colonial blue-green alga (cyanobacterium) Microcystis aeruginosa. In nonmixed eutrophic river water and cultures, O₂ evolution determinations indicated C_i limitation of photosynthesis, which was overcome either by CO₂ additions to the aqueous phase or by exposure of buoyant colonies to atmospheric CO₂. Microautoradiographs of M. aeruginosa colonies revealed partitioning of ¹⁴CO₂ fixation and photosynthate accumulation between peripheral and internal cells, particularly in large colonies. When illuminated colonies were suspended in the aqueous phase, peripheral cells accounted for at least 90% of the ¹⁴CO₂ assimilation, whereas internal cells remained unlabeled. However, when ¹⁴CO₂ was allowed to diffuse into colonies 15 min before illumination, a more uniform distribution of labeling was observed. Resultant differences in labeling patterns were most likely due to peripheral cells more exclusively utilizing CO₂ when ambient C_i concentrations were low. Among colonies located at the air-water interface, internal cells showed an increased share of photosynthate production when atmospheric ¹⁴CO₂ was supplied. This indicated that C_i transport was restricted in large colonies below the water surface, forcing internal cells to maintain a high degree of buoyancy, thus promoting the formation of surface scums. At the surface, C_i restrictions were alleviated. Accordingly, scum formation appears to have an ecological function, allowing cyanobacteria access to atmospheric CO₂ when the C_i concentration is growth limiting in the water column.     

Contribution of various carbon sources toward isoprene biosynthesis in poplar leaves mediated by altered atmospheric CO2 concentrations

Biogenically released isoprene plays important roles in both tropospheric photochemistry and plant metabolism. We performed a (13)CO(2)-labeling study using proton-transfer-reaction mass spectrometry (PTR-MS) to examine the kinetics of recently assimilated photosynthate into isoprene emitted from poplar (Populus × canescens) trees grown and measured at different atmospheric CO(2) concentrations. This is the first study to explicitly consider the effects of altered atmospheric CO(2) concentration on carbon partitioning to isoprene biosynthesis. We studied changes in the proportion of labeled carbon as a function of time in two mass fragments, M41(+), which represents, in part, substrate derived from pyruvate, and M69(+), which represents the whole unlabeled isoprene molecule. We observed a trend of slower (13)C incorporation into isoprene carbon derived from pyruvate, consistent with the previously hypothesized origin of chloroplastic pyruvate from cytosolic phosphenolpyruvate (PEP). Trees grown under sub-ambient CO(2) (190 ppmv) had rates of isoprene emission and rates of labeling of M41(+) and M69(+) that were nearly twice those observed in trees grown under elevated CO(2) (590 ppmv). However, they also demonstrated the lowest proportion of completely labeled isoprene molecules. These results suggest that under reduced atmospheric CO(2) availability, more carbon from stored/older carbon sources is involved in isoprene biosynthesis, and this carbon most likely enters the isoprene biosynthesis pathway through the pyruvate substrate. We offer direct evidence that extra-chloroplastic rather than chloroplastic carbon sources are mobilized to increase the availability of pyruvate required to up-regulate the isoprene biosynthesis pathway when trees are grown under sub-ambient CO(2).     

Stomatal density of cowpea correlates with carbon isotope discrimination in different phosphorus, water and CO2 environments

* Stomatal formation is affected by a plant's external environment, with long-distance signaling from mature to young leaves seemingly involved. However, it is still unclear what is responsible for this signal. To address this question, the relationship between carbon isotope discrimination (Delta) and stomatal density was examined in cowpea (Vigna sinensis). * Plants were grown under various environments that combined different amounts of soil phosphorus (P), soil water, and atmospheric CO(2). At harvest, stomatal density was measured in the youngest fully expanded leaf. The (13)C : (12)C ratio was measured in a young leaf to determine the Delta in mature leaves. * Results indicated that stomatal density is affected by P as well as by amounts of water and CO(2). However, stomatal responses to water and CO(2) were complex because of strong interactions with P. This suggests that the responses are relative, depending on some internal factor being affected by each external variable. Despite such complicated responses, a linear correlation was found between stomatal density and Delta across all environments examined. * It is proposed that the Delta value is a good surrogate for the long-term mean of the intercellular (C(i)) to the atmospheric (C(a)) CO(2) concentration ratio (C(i) : C(a)) and may be useful in understanding stomatal formation beyond complicated interactions.     

Comparison of Atmospheric Conditions for Culture of Clinical Specimens of Neisseria gonorrhoeae

We cultured 55 clinical specimens of Neisseria gonorrhoeae in the following atmospheric conditions: (i) 10% carbon dioxide in a CO(2) incubator; (ii) a candle extinction jar; (iii) an air convection incubator; and (iv) an anaerobic jar without added CO(2). The number and size of colonies growing on modified Thayer-Martin medium were evaluated after incubation of cultures for 24 and 48 h at 36 C. After 24 h, the specimens from the candle extinction jar had the greatest number and size of colonies, but after 48 h growth was approximately equal for specimens from the candle jar and the CO(2) incubator. Only 19 of 55 specimens grew in the air convention incubator. None of 55 clinical specimens or of 10 laboratory strains grew anaerobically. Development of colonial morphology for colony types 1, 2, 3, and 4 was studied at 24 h on a base medium that contained no hemoglobin. The relative numbers of the four colony types in specimens were comparable after 24 h of incubation in any of the three atmospheric conditions under which growth occurred, but the different types were distinguished most readily when grown in the candle extinction jar.     

RECRUITMENT OF BENTHIC MICROCYSTIS (CYANOPHYCEAE) TO THE WATER COLUMN: INTERNAL BUOYANCY CHANGES OR RESUSPENSION?1

In some lakes, large amounts of the potentially toxic cyanobacterium Microcystis overwinter in the sediment. This overwintering population might inoculate the water column in spring and promote the development of dense surface blooms of Microcystis during summer. In the Dutch Lake Volkerak, we found photochemically active Microcystis colonies in the sediment throughout the year. The most vital colonies originated from shallow sediments within the euphotic zone. We investigated whether recruitment of Microcystis colonies from the sediment to the water column was an active process, through production of gas vesicles or respiration of carbohydrate ballast. We calculated net buoyancy, as an indication of relative density, using the amounts and densities of the major cell constituents (carbohydrates, proteins, and gas vesicles). Carbohydrate content of benthic Microcystis cells was very low throughout the year. Buoyancy changes of benthic Microcystis were mostly a result of changes in gas vesicle volume. Before the summer bloom, net buoyancy and the amount of buoyant colonies in the sediment did not change. Therefore, recruitment of Microcystis from the sediment does not seem to be an active process regulated by internal buoyancy changes. Instead, our observations indicate that attachment of sediment particles to colonies plays an important part in the buoyancy state of benthic colonies. Therefore, we suggest that recruitment of Microcystis is more likely a passive process resulting from resuspension by wind‐induced mixing or bioturbation. Consequently, shallow areas of the lake probably play a more important role in recruitment of benthic Microcystis than deep areas.     

The synthesis of amino acids and sugars on an inorganic template from constituents of the prebiotic atmosphere

Inelastic Electron Tunnelling Spectroscopy (IETS) has been used to identify the reaction products present on an alumina surface when it is exposed to likely components of the earth's prebiotic atmosphere. The alumina barrier of Al-AlO _x -Pb tunnelling junctions have been exposed to water; aqueous ammonia; wet carbon monoxide gas and to aqueous formaldehyde vapour under normal atmospheric conditions at room temperature. The water spectrum shows strong coincidence with that of a genuine sample of formic acid. It is proposed that atmospheric CO₂ is involved in this surface catalyzed reaction. The aqueous ammonia spectrum is assigned as an amino acid species produced from ammonia, water and atmospheric carbon dioxide. This spectrum compares very closely with the tunnelling spectrum of a genuine sample of glycine. The wet carbon monoxide spectrum and the aqueous formaldehyde spectrum have been produced by an infusion doping process. These spectra of CO and aqueous formaldehyde are assigned as a sugar like polymer or a sugar formed on the alumina surface. A tunnelling spectrum of D(–) fructose has been produced to aid this assignment. The role of an inorganic template such as alumina in the original prebiotic synthesis of amino acids and sugars is considered.     

Distribution of microorganisms lysing Pseudomonas aeruginosa in reservoirs and soil

Various natural habitats were found to contain microorganisms producing lytic spots around their own colonies when grown on a lawn of viable Pseudomonas aeruginosa cells at 29 and 45 degrees C. The incidence of such microorganisms in water and soil was studied in quantitative terms. Contaminated waters with predominating Gram-negative heterotrophs and a high number of pseudomonades were shown to be an optimal source for the isolation of microorganisms causing the lysis of P. aeruginosa growing at 29 degrees C. Microorganisms responsible for the lysis of P. aeruginosa at 45 degrees C are abundant in the soil of mixed and foliage forests.     

Internal conductance to CO(2) diffusion and C(18)OO discrimination in C(3) leaves

(18)O discrimination in CO(2) stems from the oxygen exchange between (18)O-enriched water and CO(2) in the chloroplast, a process catalyzed by carbonic anhydrase (CA). A proportion of this (18)O-labeled CO(2) escapes back to the atmosphere, resulting in an effective discrimination against C(18)OO during photosynthesis (Delta(18)O). By constraining the delta(18)O of chloroplast water (delta(e)) by analysis of transpired water and the extent of CO(2)-H(2)O isotopic equilibrium (theta(eq)) by measurements of CA activity (theta(eq) = 0.75-1.0 for tobacco, soybean, and oak), we could apply measured Delta(18)O in a leaf cuvette attached to a mass spectrometer to derive the CO(2) concentration at the physical limit of CA activity, i.e. the chloroplast surface (c(cs)). From the CO(2) drawdown sequence between stomatal cavities from gas exchange (c(i)), from Delta(18)O (c(cs)), and at Rubisco sites from Delta(13)C (c(c)), the internal CO(2) conductance (g(i)) was partitioned into cell wall (g(w)) and chloroplast (g(ch)) components. The results indicated that g(ch) is variable (0.42-1.13 mol m(-2) s(-1)) and proportional to CA activity. We suggest that the influence of CA activity on the CO(2) assimilation rate should be important mainly in plants with low internal conductances.     

Contrasting wetland CH4 emission responses to simulated glacial atmospheric CO2 in temperate bogs and fens

Wetlands were the largest source of atmospheric methane (CH(4) ) during the Last Glacial Maximum (LGM), but the sensitivity of this source to exceptionally low atmospheric CO(2) concentration ([CO(2) ]) at the time has not been examined experimentally. We tested the hypothesis that LGM atmospheric [CO(2) ] reduced CH(4) emissions as a consequence of decreased photosynthate allocation to the rhizosphere. We exposed minerotrophic fen and ombrotrophic bog peatland mesocosms to simulated LGM (c. 200 ppm) or ambient (c. 400 ppm) [CO(2) ] over 21 months (n = 8 per treatment) and measured gaseous CH(4) flux, pore water dissolved CH(4) and volatile fatty acid (VFA; an indicator of plant carbon supply to the rhizosphere) concentrations. Cumulative CH(4) flux from fen mesocosms was suppressed by 29% (P < 0.05) and rhizosphere pore water [CH(4) ] by c. 50% (P < 0.01) in the LGM [CO(2) ], variables that remained unaffected in bog mesocosms. VFA analysis indicated that changes in plant root exudates were not the driving mechanism behind these results. Our data suggest that the LGM [CO(2) ] suppression of wetland CH(4) emissions is contingent on trophic status. The heterogeneous response may be attributable to differences in species assemblage that influence the dominant CH(4) production pathway, rhizosphere supplemented photosynthesis and CH(4) oxidation.     

The role of the C4 pathway in carbon accumulation and fixation in a marine diatom

The role of a C(4) pathway in photosynthetic carbon fixation by marine diatoms is presently debated. Previous labeling studies have shown the transfer of photosynthetically fixed carbon through a C(4) pathway and recent genomic data provide evidence for the existence of key enzymes involved in C(4) metabolism. Nonetheless, the importance of the C(4) pathway in photosynthesis has been questioned and this pathway is seen as redundant to the known CO(2) concentrating mechanism of diatoms. Here we show that the inhibition of phosphoenolpyruvate carboxylase (PEPCase) by 3,3-dichloro-2-dihydroxyphosphinoylmethyl-2-propenoate resulted in a more than 90% decrease in whole cell photosynthesis in Thalassiosira weissflogii cells acclimated to low CO(2) (10 microm), but had little effect on photosynthesis in the C(3) marine Chlorophyte, Chlamydomonas sp. In 3,3-dichloro-2-dihydroxyphosphinoylmethyl-2-propenoate-treated T. weissflogii cells, elevated CO(2) (150 microm) or low O(2) (80-180 microm) restored photosynthesis to the control rate linking PEPCase inhibition with CO(2) supply in this diatom. In C(4) organic carbon-inorganic carbon competition experiments, the (12)C-labeled C(4) products of PEPCase, oxaloacetic acid and its reduced form malic acid suppressed the fixation of (14)C-labeled inorganic carbon by 40% to 50%, but had no effect on O(2) evolution in photosynthesizing diatoms. Oxaloacetic acid-dependent O(2) evolution in T. weissflogii was twice as high in cells acclimated to 10 microm rather than 22 microm CO(2), indicating that the use of C(4) compounds for photosynthesis is regulated over the range of CO(2) concentrations observed in marine surface waters. Short-term (14)C uptake (silicone oil centrifugation) and CO(2) release (membrane inlet mass spectrometry) experiments that employed a protein denaturing cell extraction solution containing the PEPCKase inhibitor mercaptopicolinic acid revealed that much of the carbon taken up by diatoms during photosynthesis is stored as organic carbon before being fixed in the Calvin cycle, as expected if the C(4) pathway functions as a CO(2) concentrating mechanism. Together these results demonstrate that the C(4) pathway is important in carbon accumulation and photosynthetic carbon fixation in diatoms at low (atmospheric) CO(2).     

Response of potato tuber cell division and growth to shade and elevated CO2

Plants adjust their sink-organ growth rates, development and distribution of dry matter in response to whole-plant photosynthate status. To advance understanding of these processes, potato (Solanum tuberosum L.) plants were subjected to CO(2) and light flux treatments, and early tuber growth was assessed. Atmospheric CO(2) (700 or 350 micro mol mol(-1)) and light flux (shade and control illumination) treatments were imposed at two growth stages: tuber initiation (TI) and tuber bulking (TB). Elevated CO(2) increased accumulation of total net biomass when imposed at both stages, and increased tuber growth rate by about 36 %, but did not increase the number of tubers. Elevated CO(2) increased the number of cells in tubers at both TI and TB stages, whereas shade substantially decreased the number of cells at both stages. Generally, treatments did not affect cell volume or the proportion of nuclei endoreduplicating (repeated nuclear DNA replication in the absence of cell division), but the shade treatment led to a decrease in cell volume at TB and a decrease in endoreduplication at TI. Elevated CO(2) increased, and shade decreased, glucose concentration and soluble invertase activity in the cambial zones at both TI and TB, whereas sucrose concentration and activities of glucokinase, fructokinase, cell-wall-bound invertase and thymidine kinase were unaffected. Modulation of tuber cell division was responsible for much of the growth response to whole-plant photosynthate status, and treatments affected cambial-zone glucose and soluble invertase in a pattern suggesting involvement of a glucose signalling pathway.     

C-390 as sole selective agent for isolation of Pseudomonas aeruginosa from hospital waste water

Pseudomonas aeruginosa was recovered (in numbers ranging from 10(2) to 10(5) colony-forming units per millilitre) from heavily contaminated hospital waste water when grown at 41.5 degrees C on a differential medium agar containing 9-chloro-9-(4-diethylaminophenyl)-10-phenylacridan (C-390) at a final concentration of 30 micrograms/mL. The medium appeared to be highly selective for P. aeruginosa with 95-100% of all colonies isolated from four different hospital waste waters being identified as P. aeruginosa. Many strains of P. aeruginosa isolated from hospital waste waters failed to hydrolyse casein when grown on skim milk agar and this medium appeared to restrict pigment production to only pyoverdin (detectable only under ultraviolet light). However, most strains were capable of casein hydrolysis when grown on a modified skim milk medium.     

Aerenchyma Carbon Dioxide Can Be Assimilated in Typha Iatifolia L. Leaves

Leaf structural characteristics and gas-exchange measurements were used to determine whether photosynthetic tissue of Typha Iatifolia L. (cattail) utilized CO2 from the aerenchyma gas spaces, part of an internal pathway for gas transport in this wetland species. The partial pressure of CO2 (pCO2) in these aerenchyma gas spaces can be more than 10 times atmospheric pCO2. The photosynthetic tissue occurred in structurally similar adaxial and abaxial palisades, which were distinctly separated from each other by the aerenchyma gas spaces. In each palisade there were three to four layers of tightly packed, nonchlorophyllous cells separating the photosynthetic tissue from the aerenchyma gas space. Different lines of evidence indicated that CO2 conductance in the light was significantly greater across the epidermal surface than across the internal surface of both palisades. However, at an epidermal pCO2 of 350 [mu]bars and an internal pCO2 of 820 [mu]bars, the net rates of CO2 uptake (PN) across the epidermal and internal surfaces were about equal. PN across the internal surface was greater than across the epidermal surface at higher internal pCO2. Gas space pCO2 can be greater than 820 [mu]bars in the field, and therefore, PN across the internal surface could be a significant proportion of epidermal surface PN.     

Inorganic carbon acquisition in some synurophyte algae

Some characteristics of photosynthesis of three synurophyte algae, Synura petersenii, Synura uvella and Tessellaria volvocina were investigated to determine the mechanism of inorganic carbon (C(i)) uptake. All three species were found to have no external carbonic anhydrase, no capacity for direct bicarbonate uptake and a low whole-cell affinity for C(i). The internal pH of S. petersenii determined using (14)C-benzoic acid and [2-(14)C]-5,5-dimethyloxazolidine-2,4-dione was pH 7.0-7.5, over an external pH range of 5.0-7.5. Thus, the pH difference between the cell interior of S. petersenii and the external medium was large enough, over the alga's growth range, to allow the accumulation of C(i) by the diffusive uptake of CO(2). Monitoring O(2) evolution and CO(2) uptake by suspensions of S. petersenii at pH 7.0 by mass spectrometry did not indicate a rapid uptake of CO(2), and the final CO(2) compensation concentration reached was 24 +/- 0.7 microM. Furthermore, when the cells were darkened, a brief burst of CO(2) occurred before a steady rate of dark respiration was established, suggesting a loss of CO(2) by photorespiration. An examination of the kinetics of ribulose-1,5-bisphosphate carboxylase/oxygenase in homogenates of cells of S. petersenii, S. uvella and Mallomonas papillosa showed that values of the K(m) (CO(2)) were 28.4, 41.8 and 18.2 microM, respectively. These species lack the characteristics of cells with a CO(2)-concentrating mechanism because the cell affinity for C(i) appears to be determined by the relatively high CO(2) affinity of the Rubisco of these algae.     

Elevated CO2 effects on canopy and soil water flux parameters measured using a large chamber in crops grown with free-air CO2 enrichment

An arable crop rotation (winter barley-sugar beet-winter wheat) was exposed to elevated atmospheric CO(2) concentrations ([CO(2) ]) using a FACE facility (Free-Air CO(2) Enrichment) during two rotation periods. The atmospheric [CO(2) ] of the treatment plots was elevated to 550 ppm during daylight hours (T>5°C). Canopy transpiration (E(C) ) and conductance (G(C) ) were measured at selected intervals (>10% of total growing season) using a dynamic CO(2) /H(2) O chamber measuring system. Plant available soil water content (gravimetry and TDR probes) and canopy microclimate conditions were recorded in parallel. Averaged across both growing seasons, elevated [CO(2) ] reduced E(C) by 9%, 18% and 12%, and G(C) by 9%, 17% and 12% in barley, sugar beet and wheat, respectively. Both global radiation (Rg) and vapour pressure deficit (VPD) were the main driving forces of E(C) , whereas G(C) was mostly related to Rg. The responses of E(C) and especially G(C) to [CO(2) ] enrichment were insensitive to weather conditions and leaf area index. However, differences in LAI between plots counteracted the [CO(2) ] impact on E(C) and thus, at least in part, explained the variability of seasonal [CO(2) ] responses between crops and years. As a consequence of lower transpirational canopy water loss, [CO(2) ] enrichment increased plant available soil water content in the course of the season by ca. 15 mm. This was true for all crops and years. Lower transpirational cooling due to a [CO(2) ]-induced reduction of E(C) increased canopy surface and air temperature by up to 2 °C and 0.5 °C, respectively. This is the first study to address effects of FACE on both water fluxes at canopy scale and water status of a European crop rotation.     

CO2浓度升高和施氮条件下小麦根际呼吸对土壤呼吸的贡献

依托FACE技术平台,采用稳定13C同位素技术,通过将小麦(C3作物)种植于长期单作玉米(C4作物)的土壤上,研究了大气CO2浓度升高和不同氮肥水平对土壤排放CO2的δ13C值及根际呼吸的影响.结果表明:种植小麦后土壤排放CO2的δ13C值随作物生长逐渐降低,CO2浓度升高200 μmol·mol-1显著降低了孕穗、抽穗期(施氮量为250 kg·hm-2,HN)与拔节、孕穗期(施氮量为150 kg·hm-2,LN)土壤排放CO2的δ13C值,显著提高了孕穗、抽穗期的根际呼吸比例.拔节至成熟期,根际呼吸占土壤呼吸的比例在高CO2浓度下为24％～48％ (HN)和21％ ～48％ (LN),在正常CO2浓度下为20％ ～36％ (HN)和19％～32％(LN).不同CO2浓度下土壤排放CO2的δ13C值和根际呼吸对氮肥增加的响应不同,CO2浓度与氮肥用量在拔节期对根际呼吸的交互效应显著.     

Carbon isotope discrimination and bundle sheath leakiness in three C(4) subtypes grown under variable nitrogen,water and atmospheric CO(2) supply

The changes in composition and productivity of semi-arid C(4) grassland, anticipated with rising atmospheric CO(2), will depend on soil water and nutrient availability. The interactive effects of soil resource limitation and elevated CO(2 )on these grasses, furthermore, may vary among C(4) biochemical subtypes (NADP-ME, NAD-ME, PCK) that differ in bundle sheath leakiness (Phi) responses to drought and nitrogen supply. To address C(4) subtype responses to soil resource gradients, the carbon isotope discrimination (Delta), bundle sheath leakiness (Phi), leaf gas exchange (A, g(s), c(i)/c(a)) and above-ground biomass accumulation were measured on three dominant grasses of semi-desert grassland in south-eastern Arizona. Bouteloua curtipendula (PCK), Aristida glabrata (NADP-ME) and the non-native Eragrostis lehmanniana (NAD-ME) were grown in controlled-environment chambers from seed under a complete, multi-factorial combination of present ambient (370 ppm) and elevated (690 ppm) CO(2) concentration and under high and low water and nitrogen supply. E. lehmanniana (NAD-ME) had the highest photosynthetic rate (A) and lowest Phi compared to the other two grasses when grown under low nitrogen and water availability. However, favourable water and nitrogen supply and elevated atmospheric CO(2) enhanced photosynthetic performance and above-ground biomass production of B. curtipendula (PCK) to a greater extent than in A. glabrata and E. lehmanniana. Contrary to pre vious studies, Phi and Delta in the NADP-ME subtype (A. glabrata) were most affected by changing environmental conditions compared to the other subtypes; deviations from the classic NADP-ME anatomy in Aristida could have accounted for this result. Overall, response of semi-arid grasslands to rising atmospheric CO(2) may depend more on species-specific responses to drought and nitrogen limitation than on general C(4) subtype responses.     

Induction of a high-CO2-inducible, periplasmic protein, H43, and its application as a high-CO2-responsive marker for study of the high-CO2-sensing mechanism in Chlamydomonas reinhardtii

The unicellular green alga Chlamydomonas reinhardtii can acclimate to a broad range of environmental CO(2) concentrations. We observed that the cells synthesized a specific 43 kDa protein, H43, in the periplasmic space under photoautotrophic high-CO(2) conditions. Under low-CO(2) conditions, H43 disappeared. However, H43 mRNA expression was observed even under heterotrophic low-CO(2) conditions when the cells were grown with 17.4 mM acetate in darkness. When the cells were treated with 4,4'-dithiocyanatostilbene-2,2'-disulfonate (DIDS) and mersalyl to modify cell surface proteins, H43 mRNA expression was strongly affected under both heterotrophic and photoautotrophic conditions. The H43 induction pattern in a mitochondrial respiration-deficient mutant dum-1 that lacks cytochrome c oxidase was the same, but the level was much lower than that in the wild type. Even under illumination, the dissolved CO(2) concentration in the culture rapidly increased slightly following the addition of acetate and dramatically increased even further by the inhibition of photosynthesis with DCMU. Radiotracer experiments with [U-(14)C]acetate revealed that (14)CO(2) release from cells was greater in darkness than in the light due to the great stimulation of internal CO(2) evolution, resulting in an increase in external CO(2) concentration. Strong light inhibited H43 induction and DCMU promoted the induction under photoheterotrophic low-CO(2) conditions. The results demonstrate that H43 is strictly regulated by a concentration of CO(2) resulting from respiration and photosynthesis. Our results suggest that Chlamydomonas induces high-CO(2)-responsive protein H43 by sensing the concentration of ambient CO(2) with the contribution of cell surface protein.     

Physiological characteristics of the primitive CO2 concentrating mechanism in PEPC transgenic rice

C4 plants such as maize have CO2 concentrating mechanism and higher photosynthetic efficiency than C3 plants, especially under high light intensity, high temperature and drought conditions. In recent years, due to the rapid development of transgenic technique, different transgenic rice plants with high-level expression of C4 genes have been created by the successful introduction of genes encoding the key C4 photosynthetic path enzymes PEPC, PPDK and NADP-ME through agrobacteria-mediated…     

Physiological characteristics of the primitive CO<Subscript>2</Subscript> concentrating mechanism in PEPC transgenic rice

The relationship between carbon assimilation and high-level expression of the maize PEPC in PEPC transgenic rice was studied by comparison to that in the untransformed rice, japonica kitaake. Stomatal conductance and photosynthetic rates in PEPC transgenic rice were higher than those of untransformed rice, but the increase of stomatal conductance had no statistical correlation with that of photosynthetic rate. Under high levels of light intensity, the protein contents of PEPC and CA were increased significantly. Therefore the photosynthetic capacity was increased greatly (50%) with atmospheric CO2 supply. While CO2 release in leaf was reduced and the compensation point was lowered correspondingly under CO2 free conditions. Treatment of the rice with the PEPC-specific inhibitor DCDP showed that overexpression of PEPC and enhancement of carbon assimilation were related to the stability of Fv/Fm. Labeling with 14CO2 for 20 s showed more 14C was distributed to C4 primary photosynthate asperate in PEPC transgenic rice, suggesting that there exists a limiting C4 photosynthetic mechanism in leaves. These results suggest that the primitive CO2 concentrating mechanism found in rice could be reproduced through metabolic engineering, and shed light on the physiological basis for transgenic breeding with high photosynthetic efficiency.