Sulfonic acid metabolism in the diatom Navicula pelliculosa

Sulfate uptake and the cellular levels of three sulfonic acids,cysteinolic acid, sulfopropanediol, and the plant sulfolipid,were followed using ³⁵S-sulfate labeled cells of the diatomNavicula pelliculosa in silicate starvation synchrony, sulfurdeficiency, and chase experiments. Sulfate uptake was nearly completely inhibited during silicatestarvation but was resumed 3 hr after silicate addition (synchronyinduction). Cysteinolic acid and sulfopropanediol levels decreasedrapidly after silicate addition, although an earlier decreasein cysteinolic acid was noted, possibly in response to sulfateuptake inhibition. No decrease in the sulfur-containing lipidswas noted. A metabolically active role for cysteinolic acid as a reservoirof reduced sulfur was indicated, relative to sulfopropanedioland the plant sulfolipid, on the basis of rapid utilizationand turnover during sulfur deficiency and chase experiments.It was suggested that sulfopropanediol was formed by a reductivedeamination of cysteinolic acid. ¹Present address: Department of Nutrition and Food Science,Massachusetts Institute of Technology, Cambridge, Massachusetts02139, U. S. A. (Received April 2, 1973; )     

Uptake of Nitrate by the Diatom Phaeodactylum tricornutum

Ammonium-grown cells of P. tricornutum lack ability to takeup nitrate. This ability develops during 3 h of nitrogen-deprivation;development requires concurrent photosynthesis, and is inhibitedby cycloheximide. Nitrate is accumulated by cells with a developednitrate uptake mechanism. Nitrate uptake is inhibited by 10^–5M CCCP and, in darkness, by anaerobiosis; it therefore presumablyrequires ATP which can be supplied by either photophosphorylationor oxidative phosphorylation.     

Protoplast Rotation, Cell Division and Frustule Symmetry in the Diatom Navicula bacillum

Navicula bacillum frustules, like those of many other naviculoiddiatoms, exist in two forms, cis and trans, which occur in theratio 1 cis: 2 trans. During interphase the protoplast bearsa constant relation to the orientation of the hypovalve, suchthat the nucleus lies on the same side as the first-formed halfof the hypovalve. Immediately before mitosis the plastid andnucleus migrate around the cell until the nucleus lies on theopposite side from its interphase position, where it then divides.After cytokinesis the new valves form with their primary halveson the same side as the recently divided nuclei. This and the180° rotation of the protoplast during the cell cycle explainthe 1 cis: 2 trans ratio. Navicula bacillum Ehrenb., cell symmetry, diatom systematics, plastid structure, protoplast rotation     

Uptake and Accumulation of Inorganic Carbon by a Freshwater Diatom

Colman, B. and Rotatore, C. 1988. Uptake and accumulation ofinorganic carbon by a freshwater diatom.—J. exp Bot 39:1025–1032. The mechanism of uptake of inorganic carbon and its accumulationhas been studied in the freshwater diatom Navicula pelliculosa.No external carbonic anhydrase could be detected, although itwas detected in cell extracts. The rate of photosynthetic O₂evolution, in media in the range pH 7.5–8.5, exceededthe calculated rate of CO₂ supply 2- to 5-fold, indicating thatHCO^–₃ was taken up by the cells. At an external pH of7.5, the internal pH, measured by ¹⁴C-dimethyloxazolidine-2,4-dione distribution between the cells and the medium, was pH7.6 in the light and pH 7.4 in the dark. Accumulation of inorganiccarbon was determined by the silicone oil centrifugation methodand inorganic carbon pools of 23.5 mol m^–3 were found,a concentration 21.6-fold that in the external medium. The resultsindicate an active accumulation of inorganic carbon againstpH and concentration gradients in this diatom, probably by activeHCO^–₃ uptake. Key words: Bicarbonate transport, carbon dioxide, carbonic anhydrase, CO₂ affinity, CO₂ concentrating mechanism, internal pH, Navicula pelliculosa     

Active Silicon Uptake by Wheat

The absorption of Si by wheat, Triticum aestivum L. ‘Yecora Rojo,’ followed Michaelis–Menten kinetics over a concentration range of 0.004–1.0 mM. K_m and V_max were determined using linear transformations and the calculated curve fitted the data closely. The absorption resulted in accumulation ratios of 200/1 or more. In keeping with that finding, this study also demonstrated that Si uptake by wheat is under metabolic control, being severely restricted by dinitrophenol (DNP) and potassium cyanide (KCN). Silicon uptake by wheat was not significantly affected by phosphate ions, but the chemical analog Ge exerted a direct competitive effect on Si uptake, and vice versa.     

Studies on the Biochemistry and Fine Structure of Silica Shell Formation in Diatoms. Chemical Composition of Navicula pelliculosa during Silicon-Starvation Synchrony

Changes are reported in total cellular organic carbon, nucleic acids, proteins, carbohydrates, lipids and chlorophylls during the course of silicon-starvation synchrony of Navicula pelliculosa. All constituents increased at the same rate, relative to cell number, for 30 hours of exponential growth during which silicon was depleted from the medium. Increase in cell number then stopped, but net synthesis of most components continued for a further 5 to 7 hours before ceasing. Deoxyribonucleic acids and lipids accumulated throughout the 14 hour silicon-starvation period. When silicon was resupplied, lipid synthesis ceased and organic carbon and carbohydrates decreased slightly. Net synthesis remained low during the 4 hour silicon uptake period but was resumed at higher rates as cell number began to rise. In cultures transferred to the dark 1 hour prior to readdition of silicon, total carbon, carbohydrates, and lipids decreased markedly during silicon uptake and cell separation. This was due in part to conversion of protein which maintained the protein level of the dark cells close to that of cells kept in the light. Mechanisms by which silicon starvation and reintroduction of silicon might affect rates of cellular synthesis are discussed.     

The Uptake of Nitrite by the Diatom Phaeodactylum: Interactions between Nitrite and Nitrate

Freshly harvested ammonium-grown cells of the diatom Phaeodactylumtricornutum cannot take up nitrite but they acquire the abilityto do so after a period (about 3 h) of nitrogen deprivationunder conditions that allow photosynthesis. Addition of cycloheximide(1.0 µg ml^–1) prevents development as does incubationin Na⁺ or K⁺-free medium. Nitrite uptake is dependent on thepresence of Na⁺ in the medium and is inhibited by ammonium andnitrite uptake but it is concluded that the inhibition of nitriteuptake by nitrate is not of the competitive type.     

Observations on Copulation in Navicula pupula and Amphora ovalis in Relation to the Nature of Diatom Species

Within the same water body, morphological variants of the samediatom species often coexist. Observations of such phenodemesof Navicula pupula and Amphora ovalis show that, when sexual,they either fail to attract or fail to recognize each otherand do not interbreed. Since in both species the morphologicaland reproductive differences between demes appear to be reflectedalso in ecological differences, the present species conceptin diatoms needs re-evaluation. Amphora ovalis (Ktz.) Ktz., Navicula pupula Ktz., diatom microevolution, diatom systematics, reproductive isolation     

Nitrate Utilization by the Diatom Skeletonema costatum: I. Kinetics of Nitrate Uptake

Nitrate uptake has been studied in nitrogen-deficient cells of the marine diatom Skeletonema costatum. When these cells are incubated in the presence of nitrate, this ion is quickly taken up from the medium, and nitrite is excreted by the cells. Nitrite is excreted following classical saturation kinetics, its rate being independent of nitrate concentration in the incubation medium for nitrate concentration values higher than 3 micromolar. Nitrate uptake shows mixed-transfer kinetics, which can be attributed to the simultaneous contributions of mediated and diffusion transfer. Cycloheximide and p-hydroxymercuribenzoate inhibit the carrier-mediated contribution to nitrate uptake, without affecting the diffusion component. When cells are preincubated with nitrate, the net nitrogen uptake is increased.     

Nitrate Utilization by the Diatom Skeletonema costatum: II. Regulation of Nitrate Uptake

Nitrate utilization has been characterized in nitrogen-deficient cells of the marine diatom Skeletonema costatum. In order to separate nitrate uptake from nitrate reduction, nitrate reductase activity was suppressed with tungstate. Neither nitrite nor the presence of amino acids in the external medium or darkness affects nitrate uptake kinetics. Ammonium strongly inhibits carrier-mediated nitrate uptake, without affecting diffusion transfer. A model is proposed for the uptake and assimilation of nitrate in S. costatum and their regulation by ammonium ions.     

Studies on the Biochemistry and Fine Structure of Silica Shell Formation in Diatoms. Photosynthesis and Respiration in Silicon-Starvation Synchrony of Navicula pelliculosa

Rates of photosynthesis, measured by oxygen electrode or by ¹⁴CO₂ fixation, dark respiration and ³²P-phosphate incorporation are reported for the silicon-starvation synchrony of the fresh water diatom Navicula pelliculosa. During late exponential growth the rates were consistent with increase in carbon mass. During silicon starvation, rates of carbon dioxide fixation, oxygen evolution and ³²P incorporation fell, and the saturating light intensity decreased from 27,000 lux to 5000 lux. Reintroduction of silicon led to immediate transients in all parameters studied, followed by a prolonged increase in rate of dark respiration and a gradual increase in apparent photosynthesis. During release of daughter cells, the rates of dark respiration decreased as photosynthesis and ³²P incorporation increased. These results are discussed in relation to effects of silicon on the energy metabolism of the diatom.     

CO2 Uptake and Fixation by Endosymbiotic Chemoautotrophs from the Bivalve Solemya velum

Chemoautotrophic symbioses, in which endosymbiotic bacteria are the major source of organic carbon for the host, are found in marine habitats where sulfide and oxygen coexist. The purpose of this study was to determine the influence of pH, alternate sulfur sources, and electron acceptors on carbon fixation and to investigate which form(s) of inorganic carbon is taken up and fixed by the gamma-proteobacterial endosymbionts of the protobranch bivalve Solemya velum. Symbiont-enriched suspensions were generated by homogenization of S. velum gills, followed by velocity centrifugation to pellet the symbiont cells. Carbon fixation was measured by incubating the cells with ¹⁴C-labeled dissolved inorganic carbon. When oxygen was present, both sulfide and thiosulfate stimulated carbon fixation; however, elevated levels of either sulfide (>0.5 mM) or oxygen (1 mM) were inhibitory. In the absence of oxygen, nitrate did not enhance carbon fixation rates when sulfide was present. Symbionts fixed carbon most rapidly between pH 7.5 and 8.5. Under optimal pH, sulfide, and oxygen conditions, symbiont carbon fixation rates correlated with the concentrations of extracellular CO₂ and not with HCO₃⁻ concentrations. The half-saturation constant for carbon fixation with respect to extracellular dissolved CO₂ was 28 ± 3 μM, and the average maximal velocity was 50.8 ± 7.1 μmol min⁻¹ g of protein⁻¹. The reliance of S. velum symbionts on extracellular CO₂ is consistent with their intracellular lifestyle, since HCO₃⁻ utilization would require protein-mediated transport across the bacteriocyte membrane, perisymbiont vacuole membrane, and symbiont outer and inner membranes. The use of CO₂ may be a general trait shared with many symbioses with an intracellular chemoautotrophic partner.     

Enhancement of CO(2) Uptake in Avena Coleoptiles by Fusicoccin

When Avena coleoptile segments are immersed in a solution containing H¹⁴CO₃⁻, the appearance of label in the tissue is stimulated approximately 3-fold by fusicoccin application. This effect is rapid (1-2 minutes lag time), dependent upon respiratory energy, inhibited by carbonyl cyanide m-chlorophenylhydrazone, but not appreciably altered by cycloheximide treatment. A large percentage of the cellular radioactivity is found in the form of malate. Preliminary experiments indicate that CO₂, as opposed to HCO₃⁻, is the favored species of “CO₂” taken up by the segments. These results are consistent with the notion that CO₂, presumably by virtue of its fixation and conversion to malic acid, participates in the early events associated with fusicoccin-enhanced acidification of the cell wall region leading to the stimulation of cell extension growth.     

The Uptake of 63Ni by the Diatom Phaeodactylum tricornutum

Kinetic studies of the uptake of ⁶³Ni by cultures of the diatom Phaeodactylum tricornutum Bohlin revealed that the uptake capacity of the cells depended strongly on their metabolic state. Phosphate-starved cells gave saturation-type uptake curves and had low nickel-binding capacity. This was increased markedly by the addition of phosphate to the cultures. When nickel ions were added before or together with the phosphate, the increase in nickel-binding capacity of phosphate-starved cells failed to appear more or less completely. Uptake of phosphate and formation of polyphosphate by the cells were not affected by the addition of nickel to the growth medium. The phosphate dependent synthesis of the principle involved in nickel binding was induced 15–20 h after the addition of phosphate to starved cells. Whether phosphate was directly or indirectly involved in this process could not be established.     

Active CO(2) Transport by the Green Alga Chlamydomonas reinhardtii

Mass spectrometric measurements of dissolved free ¹³CO₂ were used to monitor CO₂ uptake by air grown (low CO₂) cells and protoplasts from the green alga Chlamydomonas reinhardtii. In the presence of 50 micromolar dissolved inorganic carbon and light, protoplasts which had been washed free of external carbonic anhydrase reduced the ¹³CO₂ concentration in the medium to close to zero. Similar results were obtained with low CO₂ cells treated with 50 micromolar acetazolamide. Addition of carbonic anhydrase to protoplasts after the period of rapid CO₂ uptake revealed that the removal of CO₂ from the medium in the light was due to selective and active CO₂ transport rather than uptake of total dissolved inorganic carbon. In the light, low CO₂ cells and protoplasts incubated with carbonic anhydrase took up CO₂ at an apparently low rate which reflected the uptake of total dissolved inorganic carbon. No net CO₂ uptake occurred in the dark. Measurement of chlorophyll a fluorescence yield with low CO₂ cells and washed protoplasts showed that variable fluorescence was mainly influenced by energy quenching which was reciprocally related to photosynthetic activity with its highest value at the CO₂ compensation point. During the linear uptake of CO₂, low CO₂ cells and protoplasts incubated with carbonic anhydrase showed similar rates of net O₂ evolution (102 and 108 micromoles per milligram of chlorophyll per hour, respectively). The rate of net O₂ evolution (83 micromoles per milligram of chlorophyll per hour) with washed protoplasts was 20 to 30% lower during the period of rapid CO₂ uptake and decreased to a still lower value of 46 micromoles per milligram of chlorophyll per hour when most of the free CO₂ had been removed from the medium. The addition of carbonic anhydrase at this point resulted in more than a doubling of the rate of O₂ evolution. These results show low CO₂ cells of Chlamydomonas are able to transport both CO₂ and HCO₃⁻ but CO₂ is preferentially removed from the medium. The external carbonic anhydrase is important in the supply to the cells of free CO₂ from the dehydration of HCO₃⁻.     

A description of glucose uptake in Navicula pelliculosa (Breb) hilse including a brief comparison with an associated Flavobacterium sp.

Archives of Microbiology - Navicula pelliculosa and an associated Flavobacterium sp. were isolated from the epiphyton of Scirpus maritimus, an emergent macrophyte growing in a brackish drainage...     

Technique for Measuring 14CO2 Uptake by Soil Microorganisms In Situ

Uptake of ¹⁴CO₂ in soils due to algae or sulfur-oxidizing bacteria was examined by incubation of soil samples with gaseous ¹⁴CO₂ and subsequent chemical oxidation of biologically fixed radioactive isotope to ¹⁴CO₂ for detection with a liquid scintillation counting system. The ¹⁴CO₂ was added to the soil in the gas phase so that no alteration of the moisture or ionic strength of the soil occurred. Wet oxidation of radioactive organic matter was carried out in sealed ampoules, and the ¹⁴CO₂ produced was transferred to a phenethylamine-liquid scintillation counting system with a simply constructed apparatus. The technique is inexpensive and efficient and does not require elaborate traps since several possible interfering factors were found to have no harmful effects. Experiments in coal mine regions and in geothermal habitats have demonstrated the ecological applicability of this technique for measurement of CO₂ fixation by sulfur-oxidizing bacteria and soil algae.