Bicarbonate uptake in the marine macroalga Ulva sp. is inhibited by classical probes of anion exchange by red blood cells

We demonstrate in this work that HCO _inf3 ^sup– uptake in the marine macroalga Ulva sp. features functional resemblances to anion transport mediated by anion exchangers of mammalian cell membranes. The evidence is based on (i) competitive inhibition of photosynthesis by the classical red-blood-cell anion-exchange blockers 4,4-dinitrostilbene-2,2-disulfonate and 4-nitro-4-isothiocyanostilbene-2,2-disulfonate under conditions where HCO _inf3 ^sup– , but not CO₂, was the inorganic carbon form taken up; (ii) inhibition of HCO _inf3 ^– uptake by pyridoxal phospate, indicating the involvement of lysine residues in the binding/translocation of HCO _inf3 ^sup– ; and (iii) inhibition of HCO _inf3 ^sup– (but not of CO₂) uptake by exofacial trypsin treatments, indicating the functional involvement of a plasmalemma protein. It is suggested that HCO _inf3 ^sup– uptake mediated by such a putative anion transporter can be a fundamental step in providing inorganic carbon for the CO₂-concentrating system of marine marcoalgae in an environment where the HCO _inf3 ^sup– concentration is high, but the CO₂ concentration and rates of uncatalyzed HCO _inf3 ^sup– dehydration are low.Abbreviations CI ionorganic carbon - DIDS 4,4-diisothiocyanostilbene-2,2-disulfonate - DNDS 4,4-dinitrostilbene-2,2-disulfonate - NIDS 4-nitro-4-isothiocyanostilbene-2,2-disulfonate - PLP pyridoxal phosphate - Rubisco ribulose-1,5-bisphosphate carboxylase-oxygenase     

Inorganic carbon acquisition by red seaweeds grown under dynamic light regimes

Palmaria palmata, which is able to use HCO _inf3 ^sup– as a carbon source for photosynthesis, and Lomentaria articulata, which is dependent on diffusive uptake of dissolved CO₂, were grown under constant light and light with sunflecks designed to model wave-induced fluctuations of near-shore underwater light. Both species exhibited significantly increased stable carbon isotope discrimination (more negative values of ¹³C relative to PDB) when grown with sunflecks. More negative ¹³C values were associated with decreased growth rate of P. palmata but not of L. articulata. The contrasting effects of sunflecks on the carbon-use characteristics of the two species are discussed in terms of the energetic cost of HCO _inf3 ^sup– use and the susceptibility of CO₂ diffusion-dependent species to photoinhibition.     

Water chemistry and periphyton in an alpine wetland

Remote high elevation sites are thought to be good sites to monitor global change and anthropogenic effects on ecosystems. This study was conducted during 1987–1990 in a high elevation wetland (3593 m) located in the Green Lakes Valley, Front Range, Colorado (USA). Salix spp. was the dominant riparian species in this 2 ha. wetland. Small shallow pools (<0.5 m depth) constituted a water area of 236 m³. The major source of water during the study period was snowmelt. The wetland had a well defined outlet and inlet, although an undetermined amount of water entered as groundwater from the snow patch above. Outlet discharge was 424–460 m³ during the month of July and declined thereafter as water input from the snowpatch declined. Inlet discharge was 67% of outlet discharge. Water temperatures in the outlet were always less than 6.8°C, pH 6.0–6.3, and mean conductivity 30.8 µS cm^–1. Both NO _inf3 ^sup– and SO _inf4 ^sup–2 were higher in the inlet thanin the outlet. Dominant cations in the inlet and outlet waters were Ca⁺² Mg⁺² > K⁺ + Na⁺; dominant anions were SO _inf4 ^sup–2 HCO _inf3 ^sup– > NO _inf3 ^sup– Cl^–. Nutrient limitation by P was demonstrated once using nutrient diffusing substrata. No limitation could be shown for NO _inf3 ^sup– , HCO _inf3 ^sup– , or Fe+EDTA. Slow colonization rates of periphyton on tiles were attributed to low temperatures and/or ultraviolet radiation. However, interannual differences in biomass on tiles were as much as 300% after 35 days. A minimum of 16–54 samples would be needed to detect a significant interannual change in biomass on tiles after 35 days assuming that the extreme case for periphyton patchiness. Global climate change is likely to affect discharge and water temperature in this wetland which hill have direct and indirect affects on population dynamics and ecosystem function.     

Characterisation of carbon dioxide and bicarbonate transport during steady-state photosynthesis in the marine cyanobacterium Synechococcus strain PCC7002

Net O₂ evolution, gross CO₂ uptake and net HCO _inf3 ^su– uptake during steady-state photosynthesis were investigated by a recently developed mass-spectrometric technique for disequilibrium flux analysis with cells of the marine cyanobacterium Synechococcus PCC7002 grown at different CO₂ concentrations. Regardless of the CO₂ concentration during growth, all cells had the capacity to transport both CO₂ and HCO _inf3 ^su– ; however, the activity of HCO _inf3 ^su– transport was more than twofold higher than CO₂ transport even in cyanobacteria grown at high concentration of inorganic carbon (C_i = CO₂ + HCO _inf3 ^su– ). In low-C_i cells, the affinities of CO₂ and HCO _inf3 ^su– transport for their substrates were about 5 (CO₂ uptake) and 10 (HCO _inf3 ^su– uptake) times higher than in high-C_i cells, while air-grown cells formed an intermediate state. For the same cells, the intracellular accumulated C_i pool reached 18, 32 and 55 mM in high-C_i, air-grown and low-C_i cells, respectively, when measured at 1 mM external C_i. Photosynthetic O₂ evolution, maximal CO₂ and HCO _inf3 ^su– transport activities, and consequently their relative contribution to photosynthesis, were largely unaffected by the CO₂ provided during growth. When the cells were adapted to freshwater medium, results similar to those for artificial seawater were obtained for all CO₂ concentrations. Transport studies with high-C_i cells revealed that CO₂ and HCO _inf3 ^su– uptake were equally inhibited when CO₂ fixation was reduced by the addition of glycolaldehyde. In contrast, in low-C_i cells steady-state CO₂ transport was preferably reduced by the same inhibitor. The inhibitor of carbonic anhydrase ethoxyzolamide inhibited both CO₂ and HCO _inf3 ^su– uptake as well as O₂ evolution in both cell types. In high-C_i cells, the degree of inhibition was similar for HCO _inf3 ^su– transport and O₂ evolution with 50% inhibition occurring at around 1 mM ethoxyzolamide. However, the uptake of CO₂ was much more sensitive to the inhibitor than HCO _inf3 ^su– transport, with an apparent I₅₀ value of around 250 M ethoxyzolamide for CO₂ uptake. The implications of our results are discussed with respect to C_i utilisation in the marine Synechococcus strain.Abbreviations Chl chlorophyll - C_i inorganic carbon (CO₂ + HCO _inf3 ^su– ) - CA carbonic anhydrase - CCM CO₂-concentrating mechanism - EZA ethoxyzolamide - GA glycolaldehyde - K_1/2 concentration required for half-maximal response - Rubisco ribulose-1,5,-bisphosphate carboxylase-oxygenase D.S. is a recipient of a research fellowship from the Deutsche Forschungsgemeinschaft (D.F.G.). In addition, we are grateful to Donald A. Bryant, Department of Molecular and Cell Biology and Center of Biomolecular Structure Function, Pennsylvania State University, USA, for sending us the wild-type strain of Synechococcus PCC7002.     

The impact of NO inf3 sup− loading on the freshwater macrophyte Littorella uniflora: N utilization strategy in a slow-growing species from oligotrophic habitats

The decline and disappearance of Littorella uniflora from oligotrophic waters which have become eutrophic has been associated with shading or reduced CO₂ supply. However NO _inf3 ^sup– concentrations can reach very high levels (100–2000 mmol m^–3 compared with <1–3 in oligotrophic habitats). To investigate the impact of NO _inf3 ^sup– loading alone, plants were grown under three NO _inf3 ^sup– regimes (very low, near-natural and high). The interactive effects of NO _inf3 ^sup– and photon flux density (low and high regimes) on N assimilation and accumulation, CO₂ concentrating mechanisms, C₃ photosynthesis and growth were also examined. The results were unexpected. Increased NO _inf3 ^sup– supply had very little effect on photosynthetic capacity, crassulacean acid metabolism (CAM) or lacunal CO₂ concentrations ([CO₂]_i), although there was considerable plasticity with respect to light regime. In contrast, increased NO _inf3 ^sup– supply resulted in a marked accumulation of NO _inf3 ^sup– , free amino acids and soluble protein in shoots and roots (up to 25 mol m^–3, 30 mol m^–3 and 9 mg g^–1 fresh weight respectively in roots), while fresh weight and relative growth rate were reduced. Total N content even under the very low NO _inf3 ^sup– regime (1.6–2.3%) was mid-range for aquatic and terrestrial species (and 3.1–4.3% under the high NO _inf3 ^sup– regime). These findings, together with field data, suggest that L. uniflora is not growth limited by low NO _inf3 ^sup– supply in natural oligotophic habitats, due not to an efficient photosynthetic nitrogen use but to a slow growth rate, a low N requirement and to the use of storage to avoid N stress. However the increased NO _inf3 ^sup– concentrations in eutrophic environments seem likely have detrimental effects on the long-term survival of L. uniflora, possibly as a consequence of N accumulation.     

Ulva rigida (Ulvales,Chlorophyta) tank culture as biofilters for dissolved inorganic nitrogen from fishpond effluents

Ulva rigida was cultivated in 7501 tanks at different densities with direct and continuous inflow (at 2, 4, 8 and 12 volumes d^–1) of the effluents from a commercial marine fishpond (40 metric tonnes, Tm, of Sparus aurata, water exchange rate of 16 m³ Tm^–1) in order to assess the maximum and optimum dissolved inorganic nitrogen (DIN) uptake rate and the annual stability of the Ulva tank biofiltering system. Maximum yields (40 g DW m^–2 d^–1) were obtained at a density of 2.5 g FW 1^–1 and at a DIN inflow rate of 1.7 g DIN m^–2 d^–1. Maximum DIN uptake rates were obtained during summer (2.2 g DIN M^–2 d^–1), and minimum in winter (1.1 g DIN m^–2 d^–1) with a yearly average DIN uptake rate of 1.77 g DIN m^–2 d^–1 At yearly average DIN removal efficiency (2.0 g DIN m^–2 d^–1, if winter period is excluded), 153 m² of Ulva tank surface would be needed to recover 100% of the DIN produced by 1 Tm of fish.Abbreviations DIN= dissolved inorganic nitrogen (NH _inf4 ^sup+ + NO _inf3 ^sup– + NO _inf2 ^sup– ); - FW= fresh weight; - DW= dry weight; - PFD= photon flux density; - V= DIN uptake rate     

Selection of soybean plant leaves which yield mesophyll cell isolates with maximal rates of CO2 and NO inf2 sup− photoassimilation

A problem often encountered when assaying mesophyll cell isolates prepared from mature soybean leaves, was that of poor reproducibility in rates of net ¹⁴CO₂ photoassimilation and NO₂ ^– photoreduction. It was known that soybean source leaves repeatedly displayed their most active net CO₂ photoassimilation in the period from attainment of maximal leaf area to approximately two to five days subsequent to that point. Advantage was taken of the fact that when soybean leaflets of each leaf reach their maximal area they also have reached their maximal leaf length from base to tip. This facilitates a more rapid determination of the point in time in which leaflet areas had reached A_max. Soybean plants (Glycine max cv. Williams) were propagated in the growth chamber with a 12 h light-12 h dark cycle, 25C, 65% RH, and 700 microeinsteins per meter squared per second. At 24 d post-emergence, the third leaf (numbered acropetally from the unifoliates) of each plant had just attained maximum leaflet areas (110 cm²) and lengths (13 cm). For this study, leaf mesophyll cells were enzymatically isolated, using commercially prepared pectinase, from leaflet sets of leaves selected from each of the second, third, and fourth leaf positions. Maximal rates of net ¹⁴CO₂ photoassimilation (with 5 mM HCO₃ ^–) for the second, third and fourth leaf (leaflet) isolates were, respectively, 27.0, 57.0, and 41.7 mol ¹⁴CO₂ assimilated per milligram chlorophyll per hour; simultaneously maximal rates of NO _inf2 ^sup– photoreduction (1 mM NO _inf2 ^sup– ) were, respectively, 4.4, 8.1, and 0.0 mol NO _inf2 ^sup– reduced per milligram chlorophyll per hour. These studies made it clear that in order repeatedly to attain reproducible maximal rates of leaf cell isolate net ¹⁴CO₂ photoassimilation and NO _inf2 ^sup– photoreduction, it always was necessary to select the newest, fully expanded leaves (e.g. leaf number 3) for cell isolation. Leaves from several plants only were pooled if they were excised from identically the same node on each of the plants.Abbreviations A_max - maximum leaflet (trifoliolate) area attained during ontogeny - CO₂ - CO₂ gas dissolved in solution - HCO _inf3 ^sup– - bicarbonate - L_max - maximum leaf blade length (midvein) attained during ontogeny - NiRase - chloroplast nitrite reductase (reduced ferredoxin) - NiPR - nitrite photoreduction - PE - post-emergence - Pn - net CO₂ photoassimilation (for leaflets and mesophyll cell isolates) - PPRC - pentose phosphate reductive cycle     

Uptake of technetium by freshwater green microalgae

Summary Accumulation of [⁹⁹Tc]pertechnetate ions (⁹⁹TcO₄ ^–) by the freshwater microalgae Chlorella emersonii, Chlamydomonas reinhardtii and Scenedesmus obliquus has been characterized. In all three species, accumulation consisted of a single rapid energy-independent phase (biosorption), and no energy-dependent accumulation was observed. Biosorption of ⁹⁹TcO _inf4 ^sup– by all three species was concentration dependent, followed a Freundlich adsorption isotherm, and was dependent on pH with increased accumulation by cells with decreasing external pH. Elevated external NaCl concentrations also caused increased accumulation of ⁹⁹TcO _inf4 ^sup– by the cells, as did increased external osmotic potential. Concentrations of K⁺, Mg²⁺ and Ca²⁺ increased accumulation of ⁹⁹TcO _inf4 ^sup– , but concentrations of HCO _inf3 ^sup– , SO _inf4 ^sup2– and CO _inf3 ^sup2– decreased ⁹⁹TcO _inf4 ^sup– accumulation by the cells. Most of the ⁹⁹TcO _inf4 ^sup– accumulated by all three species was easily desorbed by 10 mm buffers at various pH values, 0.5 m NaCl, 10 mm Na₂CO₃ or 10 mm Na₂SO₄. No differences in the amount of desorption were observed between the various desorption agents used. Correspondence to: G. M. Gadd     

Biological denitrification of water in a two-chambered immobilized-cell bioreactor

Summary A double-chambered bioreactor based on a composite immobilized-cell gel layer/microporous membrane structure was applied to the continuous denitrification of high-nitrate water. Immobilized denitrifying bacteria (Pseudomonas denitrificans) were provided with separate flows of nitrate and carbon (C) nutrient, with no contamination of the treated water by cell leakage from the gel. Using acetate (7.5 mm) as a C source and a C/N ratio of 3 (mol/mol), specific denitrification rates ranging from 15 to 25 g NO _inf3 ^sup– · h^–1 · – cm^–2 membrane surface (50–85 g NO _inf3 ^sup– · h^–1 · cm^–3 gel) were obtained. The denitrifying activity remained stable for several months. At the flow rate used (10 cm³ · h^–1), the effluents contained noticeable amounts of NO _inf2 ^sup– ions but the treated water remained uncontaminated by the carbon nutrient. Most NO _inf2 ^sup– ions disappeared from the treated water in a second reactor connected in series. When fed with an unchlorinated sludge supernatant as C nutrient, immobilized bacteria performed efficient denitrification of water for only 3 weeks. Diffusion experiments showed that acetate ions diffused much less rapidly than NO _inf3 ^sup– or NO _inf2 ^sup– ions through the composite structure. Further developments of the system are considered.     

Diurnal variation in physical-chemical properties and primary production in the interconnected marine,mangrove and freshwater biotopes of Kakinada coast,Andhra Pradesh,India

Diurnal variation in hydrological variables and dissolved inorganic nutrients such as PO _inf4 ^sup3– -P, N O _inf2 ^sup– -N, NO _inf3 ^sup– -N and NH _inf4 ^sup+ -N were studied in three interconnected biotopes including freshwater, marine and mangrove brackish water of the Kakinada coastal zone, Andhra Pradesh. Samples were collected at intervals of 3 hours, for a period of 24 hours. In the marine environment salinity varied from 26 to 32 whereas in the mangrove waters it fluctuated from 12 to 20 and in both biotopes salinity showed bimodal type of oscillation. Dissolved oxygen content was high in the mangrove waters during day time but decreased rapidly during the night hours. In the marine environment PO_f4 ^p3–-P concentration varied from 0.345 to 1.195 g at l^–1, NO _inf3 ^sup– -N from 1.03 to 6.62 g at l^–1 and NO _inf2 ^sup– -N from 0.086 to 0.506 g at l^–1. The highest and the lowest concentrations of PO _inf4 ^sup3– -P, NO _inf3 ^sup– -N, NO _inf2 ^sup– -N recorded in the mangrove waters were 0.790 and 0.325 g at l^–1, 7.10 and 1.60 g at l^–1 and 0.278 and 0.060 g at l^–1, respectively. The concentration of PO _inf4 ^sup3– -P, NO _inf3 ^sup– -N and NO _inf2 ^sup– -N were high in the freshwater canal, the maximum and minimum values being 1.110 and 0.730 g at l^–1, 26.40 and 9.98 g at l^–1 and 0.520 and 0.252 g at l^–1 respectively. The concentration of ammonia was relatively high in the mangrove water. Gross and net primary production in the mangrove water was 4 times higher than in the marine biotope. There was no export of dissolved nutrients from the mangrove environment to the adjacent marine waters.     

Inorganic-carbon transport in some marine eukaryotic microalgae

Inorganic-carbon transport was investigated in the eukaryotic marine microalgaeStichococcus minor, Nannochloropsis oculata and aMonallantus sp. Photosynthetic O₂ evolution at constant inorganic-carbon concentration but varying pH showed thatS. minor had a greater capacity for CO₂ rather than HCO ₃ ^– utilization but forN. oculata andMonallantus HCO ₃ ^– was the preferred source of inorganic carbon. All three microalgae had a low affinity for CO₂ as shown by the measurement of inorganic-carbon-dependent photosynthetic O₂ evolution at pH 5.0. At pH 8.3, where HCO ₃ ^– is the predominant form of inorganic carbon, the concentration of inorganic carbon required for half-maximal rate of photosynthetic O₂ evolution [K _0.5 (CO₂)] was 53 M forMonallantus sp. and 125 M forN. oculata, values compatible with HCO ₃ ^– transport. Neither extra- nor intracellular carbonic anhydrase was detected in these three microalgal species. It is concluded that these microalgae lack a specific transport system for CO₂ but that HCO ₃ ^– transport occurs inN. oculata andMonallantus, and in the absence of intracellular carbonic anhydrase the conversion of HCO ₃ ^– to CO₂ may be facilitated by the internal pH of the cell.     

Energetics and regulations of formate and hydrogen metabolism by Methanobacterium formicicum

Accumulation of formate to millimolar levels was observed during the growth of Methanobacterium formicicum species on H₂–CO₂. Hydrogen was also produced during formate metabolism by M. formicicum. The amount of formate accumulated in the medium or the amount H₂ released in gas phase was influenced by the bicarbonate concentration. The formate hydrogenlyase system was constitutive but regulated by formate. When methanogenesis was inhibited by addition of 2-bromoethane sulfonate, M. formicicum synthesized formate from H₂ plus HCO _inf3 ^sup- or produced H₂ from formate to a steady-state level at which point the Gibbs free energy (G) available for formate synthesis or H₂ production was approximately -2 to -3 kJ/reaction. Formate conversion to methane was inhibited in the presence of high H₂ pressure. The relative rates of conversion of formate and H₂ were apparently controlled by the G available for formate synthesis, hydrogen production, methane production from formate and methane production from H₂. Results from ¹⁴C-tracer tests indicated that a rapid isotopic exchange between HCOO^- and HCO _inf3 ^sup- occurred during the growth of M. formicicum on H₂–CO₂. Data from metabolism of ¹⁴C-labelled formate to methane suggested that formate was initially split to H₂ and HCO _inf3 ^sup- and then subsequently converted to methane. When molybdate was replaced with tungstate in the growth media, the growth of M. formicicum strain MF on H₂–CO₂ was inhibited although production of methane was not Formate synthesis from H₂ was also inhibited.     

The ability of several high arctic plant species to utilize nitrate nitrogen under field conditions

The ability to utilize NO _inf3 ^sup– in seven high arctic plant species from Truelove Lowland, Devon Island, Canada was investigated, using an in vivo assay of maximum potential nitrate reductase (NR) activity and applications of ¹⁵N. Plant species were selected on the basis of being characteristic of nutrient-poor and nutrient-rich habitats. In all species leaves were the dominant site of NR activity. Root NR activity was negligible in all species except Saxifraga cernua. NO _inf3 ^sup– availability per se did not appear to limit NR activity of the species typically found on nutrient-poor sites (Dryas integrifolia, Saxifraga oppositifolia, and Salix arctica), or in Cerastium alpinum, as leaf NR activities remained low, even after NO _inf3 ^sup– addition. ¹⁵NO _inf3 ^sup– uptake was limited in D. integrifolia and Salix arctica. However, the lack of field induction of NR activity in C. alpinum and Saxifraga oppositifolia was not due to restricted nitrate uptake, as ¹⁵NO _inf3 ^sup– labelled NO _inf3 ^sup– entered the roots and shoots of both species. Leaf NR activity rates were low in three of the species typical of nutrient-rich habitats (O. digyna, P. radicatum and Saxifraga cernua), sampled from a site containing low soil NO _inf3 ^sup– . Additions of NO _inf3 ^sup– significantly increased leaf NR activity in these latter species, suggesting that potential NR activity was limited by the availability of NO _inf3 ^sup– . ¹⁵N labelled NO _inf3 ^sup– was taken up by O. digyna. P. radicatum and Saxifraga cernua. Although two species (D. integrifolia and Salix arctica) showed little utilization of NO _inf3 ^sup– , we concluded that five of the seven selected high arctic plant species (C. alpinum, O. digyna, P. radicatum, Saxifraga cernua and Saxifraga oppositifolia) do have the potential to utilize NO _inf3 ^sup– as a nitrogen source under field conditions, with the highest potential to utilize NO _inf3 ^sup– occurring in three of the species typically found on fertile habitats.     

The active species of “CO2” utilized in ferredoxin-linked carboxylation reactions

The active species of CO₂ , i.e. CO₂ or HCO ₃ ^– –(H₂CO₃) utilized by enzymes catalyzing ferredoxin-linked carboxylation reactions was determined. The enzyme investigated was pyruvate synthase from Clostridium pasteurianum (EC 1.2.7.1; Pyruvate: ferredoxin oxidoreductase). Data were obtained which were compatible with those expected if CO₂ is the active species.The dissociation constant (K _S) of the enzyme-CO₂ complex was measured. At pH 7.2 K _Sfor CO₂ of pyruvate synthase was found to be approximately 5 mM.Abbreviations Fd ferredoxin No distinctions are made between CO₂, H₂CO₃, HCO ₃ ^– and CO ₃ ⁼ when the symbol CO₂ is used.     

Nitrogen and phosphorus uptake by the Brazilian kelp Laminaria abyssalis (Phaeophyta) in culture

The uptake of ammonium, nitrate and phosphate by laboratory-grown young sporophytes of Laminaria abyssalis was measured in a perturbed system (batch mode) at 18 °C and 35 ± 5 µE m^–2 s^–1 photon flux density. Uptake of all appeared to follow saturation-type nutrient uptake kinetics. The NO _inf3 ^sup– (K _s = 14.0 µM, V _max = 5.0 µmol h^–1 g^–1 dry wt) and NH _inf4 ^sup+ (K _s = 4.6 µM, V _max= 2.0 µmol h^–1 g^–1 dry wt) were taken up simultaneously, although NH _inf4 ^sup+ was taken up more rapidly. Values of K ₃ and V _max for phosphate were, respectively, 2.21 µM and 0.83 µmol h^–1 g^–1 dry wt. Nitrate and phosphate were both consumed in similar rates (V _max /Ks 0.37) at low concentrations. NH _inf4 ^sup+ , thus, might be a more efficient form of N fertilizer if artificial enrichment of seawater is used.     

Short-chain fatty acids and CO2 as regulators of Na+ and Cl− absorption in isolated sheep rumen mucosa

Summary Unidirectional ²²Na⁺ and ³⁶Cl^– fluxes were determined in short-circuited, stripped rumen mucosa from sheep by using the Ussing chamber technique. In both CO₂/HCO _– ³ -containing and CO₂/HCO _– ³ -free solutions, replacement of gluconate by short-chain fatty acids (SCFA, 39 mM) significantly enhanced mucosal-toserosal Na⁺ absorption without affecting the Cl^– transport in the same direction. Short-chain fatty acid stimulation of Na⁺ transport was at least partly independent of Cl^– and could almost completely be abolished by 1 mM mucosal amiloride, while stimulation of Na⁺ transport was enhanced by lowering the mucosal pH from 7.3 to 6.5. Similar to the SCFA action, raising the PCO₂ in the mucosal bathing solution led to an increase in the amiloride-sensitive mucosal-to-serosal Na⁺ flux. Along with its effect on sodium transport, raising the PCO₂ also stimulated chloride transport. The results are best explained by a model in which undissociated SCFA and/or CO₂ permeate the cell membrane and produce a raise in intracellular H⁺ concentration. This stimulates an apical Na⁺/H⁺ exchange, leading to increased Na⁺ transport. The stimulatory effect of CO₂ on Cl^– transport is probably mediated by a Cl^–/HCO _– ³ exchange mechanism in the apical membrane. Binding of SCFA anions to that exchange as described for the rat distal colon (Binder and Mehta 1989) probably does not play a major role in the rumen.Abbreviations DIDS 4,4'-diisothiocyanatostilbene-2,2'-disulfonic acid - G _t transepithelial conductance (mS·cm^-2) - HSCFA undissociated short-chain fatty acids - J _ms mucosal-to-serosal flux (Eq · cm^-2 · h^-1) - J _net net flux (Eq · cm^-2 · h^-1) - J _sm serosal-to-mucosal flux (Eq · cm^-2 · h^-1) - PD transepithelial potential difference (mV) - SCFA dissociated short-chain fatty acids - SCFA short-chain fatty acids     

Seasonal periodicity of planktonic desmids in oligotrophic lake Grane Langsø, Denmark

60 desmids were identified from plankton samples collected during 20 years, bimonthly, or monthly in 5 of the years. Only 5 desmids were perennial, performing cell divisions in a frozen lake at PAR at only 5 cal cm ^–2 day ^–1, below ice-cover. The greatest number of desmids per 100 ml lake water was found in September; it never exceeded 150. Owing to a concentration of free CO₂ of only 0.02 mM l^–1 and a HCO _inf3 ^sup– concentration of 0.002 mM l^–1 the densities of desmid cells were often smaller than 1 cell per 100 ml lake water. A density of 1 cell per 1 ml was attained only by Staurastrum longipes, a desmid showing temporal variation. The relation of the compound phytoplankton quotient to two components of the CO₂-system is discussed. Three new taxa are described: Staurastrum brachiatum Ralfs var. bicorne n. var., Staurastrum thomassonii n.sp. and Xanthidium antilopaeum Kütz. f. bimaculatum n.f..     

Pathways for bicarbonate transfer across the serosal membrane of turtle urinary bladder: Studies with a disulfonic stilbene

Summary Bicarbonate is transferred across the serosal (S) membrane of the epithelial cells of the turtle bladder in two directions. Cellular HCO ₃ ^– generated behind the H⁺ pump moves across this membrane into the serosal solution. This efflux of HCO ₃ ^– is inhibited by SITS (4-isothiocyano-4-acetamido-2,2-disulfonic stilbene). When HCO ₃ ^– is added to the serosal solution it is transported across the epithelium in exchange for absorbed Cl^–. This secretory HCO ₃ ^– flow traverses the serosal cell membrane in the opposite direction. In this study the effects of serosal addition of 5×10^–4 m SITS on HCO ₃ ^– secretion and Cl^– absorption were examined. The rate of H⁺ secretion was brought to zero by an opposing pH gradient, and 20mm HCO ₃ ^– was added toS. HCO ₃ ^– secretion, measured by pH stat titration, was equivalent to the increase inMS Cl^– flux after HCO ₃ ^– addition. Neither theSM flux of HCO ₃ ^– nor theMS flux of Cl^– were affected by SITS. In the absence of electrochemical gradients, net Cl^– absorption was observed only in the presence of HCO ₃ ^– in the media; under such conditions, unidirectional and net fluxes of Cl^– were not altered by serosal or mucosal SITS. H⁺ secretion, however, measured simultaneously as the short-circuit current in ouabain-treated bladders decreased markedly after serosal SITS. The inhibition of the efflux of HCO ₃ ^– in series with the H⁺ pump and the failure of SITS to affect HCO ₃ ^– secretion and Cl^– absorption suggest that the epithelium contains at least two types of transport systems for bicarbonate in the serosal membrane.     

Developmental Change in CO2 Compensation Concentrations in Spartina alterniflora Results from Sigmoidal Photosynthetic CO2 Responses

We investigated seasonal patterns of photosynthetic responses to CO₂ concentrations in Spartina alterniflora Loisel, an aerenchymous halophyte grass, from a salt marsh of the Bay of Fundy (NB, Canada), and from plants grown from rhizome in controlled-environment chambers. From late May to August, CO₂ compensation concentrations () of field-grown leaves varied between 2.5–10.7 cm³(CO₂) m^–3, with a mean of 5.4 cm³(CO₂) m^–3. From September onwards field leaves showed CO₂ compensation concentrations from 6.6–21.1 cm³(CO₂) m^–3, with a mean of 13.1 cm³ m^–3 well into the C₃–C₄ intermediate range. The seasonal variability in did not result from changing respiration, but rather from a sigmoidal response of net photosynthetic rate (P _N) to applied CO₂ concentration, found in all tested leaves but which became more pronounced late in the season. One explanation for the sigmoidal response of P _N to external CO₂ concentration could be internal delivery of CO₂ from roots and rhizomes to bundle sheath cells via the aerenchyma, but the sigmoidal responses in S. alterniflora persisted out to the tips of leaves, while the aerenchyma extend only to mid-leaf. The sigmoidicity persisted when CO₂ response curves were measured from low to high CO₂, or from high to low CO₂, and even when prolonged acclimation times were used at each CO₂ concentration.