Effects of Variations in Daylength and Temperature on Net Rates of Photosynthesis, Dark Respiration, and Excretion by Isochrysis galbana Parke

The effects of variable daylength and temperature on net rates of photosynthesis, dark respiration, and excretion of a unicellular marine haptophyte, Isochrysis galbana Parke, were examined and related to division rates. Six combinations of daylength (18:6, 12:12, 6:18 light:dark, LD) and temperature (20, 25 C) were used. Daily rates of net photosynthesis were closely correlated to division rates, suggesting a direct relationship, and were maximal when cells were grown at 12:12 LD at both temperatures and 18:6 LD at 20 C. A daylength of 6 hours decreased daily rates by decreasing the time for carbon uptake. Further, cells grown with this daylength had maximal chlorophyll a contents, suggesting a physiological adaptation by photosynthetic units to short light periods. A photoperiod of 18:6 LD at 25 C decreased daily rates of net photosynthesis by reducing the hourly rate of net photosynthesis via an unidentified mechanism. The importance of rates of net dark respiration in controlling daily net photosynthesis was small, with carbon lost during dark periods varying between 4 and 14% of that gained during light periods. Also, the influence of net excretion was small, varying between 1.0 and 5.5% of daily net photosynthesis.     

EFFECTS OF SMALL-SCALE TURBULENCE ON PHOTOSYNTHESIS,PIGMENTATION, CELL DIVISION,AND CELL SIZE IN THE MARINE DINOFLAGELLATE GOMAULAX POLYEDRA (DINOPHYCEAE)1

Several experiments were conducted to understand better the physiological mechanisms underlying growth inhibition of the dinoflagellate Gonyaulax polyedra Stein due to small-scale turbulence shear. To measure photosynthetic ¹⁴C uptake, a “phytoplankton wheel” device for rotating cultures in closed bottles was used. Turbulence was quantified biologically in the bottles by comparing growth inhibition with that in cultures with constant shear between a fixed cylinder and an outer concentric rotating cylinder (a stable Couette flow). At saturating irradiances, particulate photosynthesis (P_sat) or photosynthesis per unit chlorophyll (P^B_sat) were not inhibited completely at the highest turbulence level (26.6 rad.s^?1), and photosynthesis was less sensitive than growth. Photosynthesis per cell (P^C_sat) was increased by turbulence. In three experiments on the effects of turbulence on photosynthesis versus irradiance curves, the slope of the curve, α, for particulate photosynthesis at limiting irradiances did not change. Photosynthesis per unit chlorophyll per unit irradiance (α^B) decreased at high (but not intermediate) turbulence levels. Photosynthesis per cell per unit irradiance, α^C, increased with turbulence, suggesting an increase in photosynthetic efficiency in turbulent cultures. In two of the three experiments, respiration rates increased with turbulence, and in one experiment excretion of photosynthetically fixed ¹⁴C was not affected by motion. Ratios of accessory pigments to chlorophyll a did not change with turbulence, but pigments per cell and per dry weight increased with turbulence. These findings suggest little or no disruption of the photosynthetic apparatus. When turbulence was applied for 1 week, β-carotene increased while peridinin and diadinoxanthin decreased, suggesting inhibition of synthesis of these latter pigments by prolonged turbulence. Since cell numbers did not increase or decreased during turbulent 72–h incubations, cell division was inhibited and also the cells were very much enlarged. Increases in α^C per cell suggest that, in the sea, photo synthetic metabolism can persist efficiently without cell division during turbulent episodes. After turbulence ceases or reaches low levels again, cells can then divide and blooms may form. Thus, blooms can come or go fairly rapidly in the ocean depending on the degree of wave- and wind-induced turbulence.     

The effect of nutrient deficiencies on phosynthesis and respiration in spinach

Summary Spinach plants were grown in nutrient-culture solutions containing reduced levels of all the macro- and micro-nutrient elements except cobalt and chlorine. The rates of photosynthesis (carbon dioxide fixation in the light expressed on a per unit chlorophyll or per unit fresh-weight basis) and respiration (carbon dioxide evolution in the dark expressed on a per unit nitrogen or per unit fresh-weight basis) for whole plants were measured using infra-red gas analysis techniques. Measurements were made when the plants displayed clear symptoms of deficiency relative to control plants. All nutrient deficiencies except iron and molybdenum depressed photosynthesis when chlorophyll was the basis of calculation; manganese-, copper-, phosphorus- and potassium-deficient plants showed the greatest depression. Alternatively when photosynthesis was calculated on a fresh weight basis calcium was the only deficiency which had no affect. Similarly when respiration was calculated on a nitrogen basis all deficiencies except iron, molybdenum and nitrogen result in depressed rates but when respiration was expressed on a fresh-weight basis potassium deficiency resulted in enhanced respiration rates and nitrogen, phosphorus, sulphur, manganese, zinc and molybdenum deficiencies resulted in reduced respiration rates.     

EFFECT OF COPPER ON GROWTH,SILICIC ACID UPTAKE AND SOLUBLE POOLS OF SILICIC ACID IN THE MARINE DIATOM,THALASSIOSIRA WEISFLOGII (BACILLARIOPHYCEAE)1

The toxicity of Cu to Thalassiosira weissflogii (Grunow) was investigated, focusing on the internal soluble pool of silicic acid. Silicic acid uptake and growth rates were found to be functions of both the cupric ion activity and the concentration of silicic acid in the growth medium. The soluble pool of Si per cell depended on the balance between the uptake rate and the division rate. The soluble pool in non-dividing cultures reflected simply the uptake rate (and inhibition by copper of the uptake rate), but in dividing cultures the soluble pools had complex patterns with time depending on uptake rates and timing of division. Intracellular soluble pools of silicic acid are a good indicator for the relative inhibition of uptake and growth processes.     

Carbon dynamics of logarithmetic and stationary phase phytoplankton as determined by track autoradiography

Track autoradiographic analysis of photosynthetic radiocarbon incorporation at the cellular level indicated that the carbon uptake rate and carbon pool size of exponentially growing (log phase) Scenedesmus cells was threefold that of stationary phase cells, while carbon turnover rates were similar. Carbon fixation was uncoupled from growth and cell division in the stationary phase cells, which were larger and contained less chlorophyll per unit volume than log phase cells. Changes in the temporal pattern of isotope incorporation were evident at the cell level prior to the cessation of division and transition to stationary phase, while bulk carbon fixation responded only the second day after cell division ceased. The carbon uptake patterns of a marine nanoplankter from a nutrient-enriched natural sample resembled that of log phase cells while the control population pattern resembled that of stationary cells. The physical, biochemical, and metabolic differences between log and stationary phase cells are potentially measurable by flow cytometry procedures currently in use and under development. The use of flow cytometry to sort cell types for analysis by track autoradiography and subsequent correlation of metabolic characteristics with flow cytometry signatures is a feasible means of investigating the heterogeneity of phytoplankton metabolic state in the marine environment.     

Effects of elevated CO2 on growth,calcification, and spectral dependence of photoinhibition in the coccolithophore Emiliania huxleyi (Prymnesiophyceae)1

We studied the effects of elevated CO₂ concentrations on cell growth, calcification, and spectral variation in the sensitivity of photosynthesis to inhibition by solar radiation in the globally important coccolithophore Emiliania huxleyi. Growth rates and chlorophyll a content per cell showed no significant differences between elevated (800 ppmv) and ambient (400 ppmv) CO₂ conditions. However, the production of organic carbon and the cell quotas for both carbon and nitrogen, increased under elevated CO₂ conditions, whilst particulate inorganic carbon production rates decreased under the same conditions. Biometric analyses of cells showed that coccoliths only presented significant differences due to treatments in the central area width. Most importantly, the size of the coccosphere decreased under elevated CO₂ conditions. The susceptibility of photosynthesis to inhibition by ultraviolet radiation (UVR) was estimated using biological weighting functions (BWFs) and a model that predicts photosynthesis under photosynthetically active radiation and UVR exposures. BWF results demonstrated that the sensitivity of photosynthesis to UVR was not significantly different between E. huxleyi cells grown under elevated and present CO₂ concentrations. We propose that the acclimation to elevated CO₂ conditions involves a physiological mechanism of regulation and allocation of energy and metabolites in the cell, which is also responsible for altering the sensitivity to UVR. In coccolithophores, this mechanism might be affected by the decrease in the calcification rates.     

Inhibition of cell division in Escherichia coli K-12 by the R-factor R1 and copy mutants of R1.

The effect of the copy number of plasmid R1drd-19 on cell division of Escherichia coli K-12 was studied in populations growing as steady-state cultures at different growth rates, the growth rate being varied by use of different carbon sources. The plasmid copy number was also varied by using copy mutants of the R-factor. The mean cell size was larger in populations carrying an R-factor than in R-factorless populations, an effect that was more pronounced at low growth rates and in populations carrying R-factor copy mutants. The increased cell size was due to formation of elongated cells in a fraction of the population and to an increase in the diameter of all cells. The majority of the cells divided at a normal cell length, but the presence of an R-factor caused some cells to elongate, probably by the uncoupling of chromosome replication and cell division. This can be explained as a competition between the chromosome and plasmid replicons for some replication factor(s), presumably acting on both initiation and elongation of replication. The formation of elongated cells was a reversible process, but occasionally some of the elongated cells reached lengths 20 times that of newborn cells. If cell division did not occur at the normal cell size, the septum was not formed until the cell size was four times that of a newborn cell. When an elongated cell divided, it usually formed a polar septum, thus producing a newborn cell of normal cell length. The ability of plasmid-containing cells to omit one cell division but to retain the capacity of dividing one mass doubling later is compatible with a mechanical model for septum formation and cell division.     

SILICON METABOLISM IN DIATOMS. VI. SILICIC ACID UPTAKE BY A COLORLESS MARINE DIATOM,NITZSCHIA ALBA LEWIN AND LEWIN12

Cells of the colorless, heterotrophic diatom Nitzschia alba, after removal from the culture medium, were able to absorb silicic acid from a salt solution lacking carbon, nitrogen, and phosphorus. Silicic acid uptake continued for approximately 12 hr. At low cell densities (ca. 2.5×10⁵ cells/ml), the cell number doubled under these conditions. At high cell densities (ca. 2 ×10⁶ cells/ml) no cell division resulted. When such cells were washed with a salt solution, their ability to absorb silicic acid was somewhat impaired. The degree of impairment became progressively more pronounced after subsequent washing treatments. A heat-stable factor washed from the cells and present in the first “wash water” was able to restore completely the ability of washed cells to absorb silicic acid. The factor was not identified. Aspartic acid (5 ± 10^-4 M) or glutamine (5 ±10^-4 M) when added to the saline solution similarly promoted complete recovery. A t such concentrations, these substances had only a slight effect on unwashed cells. A solution of 5 ±10^-4 M Na glutamate (or aspartic acid plus glutamine) had an men more pronounced effect, and in addition promoted cell division or growth" of unwashed cells. Several other amino acids and other compounds tested were apparently without effect.     

Effects of diazepam on photosynthesis, respiration, rubidium uptake, and finestructure of Scenedesmus obliquus in synchronous cultures.

Effects of diazepam (Valium) on photosynthesis, chlorophyll/photosynthesis ratios, respiration, uptake of rubidium ions, and ultrastructure of Scenedesmus obliquus synchronized by a light-dark regimen of 14:10 hrs were determined. 80 and 160 muM diazepam, added to the nutrient medium at the start of the light-dark change (i.e., start of the cell cycle) gradually reduced rates of photosynthesis, below the initial rates from the beginning of the experiment. Contents of chlorophyll, however, remained nearly unaffected. Consequently, the diazepam-treated cells had a higher chlorophyll/photosynthesis ratio--also with regard to respiration in order to calculate the gross photosynthesis. The occurrence of photorespiration cannot be assumed. The net influx of rubidium was slightly reduced by 100 muM diazepam 0.5 and 2.0 hrs after the start of the cell cycle and was strongly inhibited after 5 to 14 hrs. 80 and 160 muM diazepam caused separation of thylakoids, formation of giant mitochondria and enlargement of vacuoles.     

Effects of organic carbon sources on growth, photosynthesis, and respiration of Phaeodactylum tricornutum

The growth, photosynthesis, and respiration of the marine diatom Phaeodactylum tricornutum were examined under photoautotrophic and mixotrophic conditions. 100 mM glycerol, acetate, and glucose significantly increased specific growth rate, and mixotrophic growth achieved higher biomass concentrations. Under mixotrophic conditions, respiration rate (R _d) and light compensation irradiance (I _c) were significantly higher, but net maximum photosynthetic O₂ evolution rate (P _m) and saturation irradiance (I _k) were depressed. Organic carbon sources decreased the cell photosynthetic pigment content and chlorophyll a to c ratio, but with a higher carotenoid to chlorophyll a ratio. Ratios of variable to maximum chlorophyll fluorescence (F _v/F _m) and 77 K fluorescence spectra of mixotrophic cells indicated a reduced photochemical efficiency of photosystem II. The results were accompanied by lower electron transport rate. Therefore, organic carbon sources reduced the photosynthesis efficiency, and the enhancement of biomass of P. tricornutum implied that organic carbon sources had more pronounced effects on respiration than on photosynthesis.     

Guard cell photosynthesis is critical for stomatal turgor production,yet does not directly mediate CO2‐ and ABA‐induced stomatal closing

Stomata mediate gas exchange between the inter‐cellular spaces of leaves and the atmosphere. CO₂ levels in leaves (Ci) are determined by respiration, photosynthesis, stomatal conductance and atmospheric [CO₂]. [CO₂] in leaves mediates stomatal movements. The role of guard cell photosynthesis in stomatal conductance responses is a matter of debate, and genetic approaches are needed. We have generated transgenic Arabidopsis plants that are chlorophyll‐deficient in guard cells only, expressing a constitutively active chlorophyllase in a guard cell specific enhancer trap line. Our data show that more than 90% of guard cells were chlorophyll‐deficient. Interestingly, approximately 45% of stomata had an unusual, previously not‐described, morphology of thin‐shaped chlorophyll‐less stomata. Nevertheless, stomatal size, stomatal index, plant morphology, and whole‐leaf photosynthetic parameters (PSII, qP, qN, F_V′/F_M′) were comparable with wild‐type plants. Time‐resolved intact leaf gas‐exchange analyses showed a reduction in stomatal conductance and CO₂‐assimilation rates of the transgenic plants. Normalization of CO₂ responses showed that stomata of transgenic plants respond to [CO₂] shifts. Detailed stomatal aperture measurements of normal kidney‐shaped stomata, which lack chlorophyll, showed stomatal closing responses to [CO₂] elevation and abscisic acid (ABA), while thin‐shaped stomata were continuously closed. Our present findings show that stomatal movement responses to [CO₂] and ABA are functional in guard cells that lack chlorophyll. These data suggest that guard cell CO₂ and ABA signal transduction are not directly modulated by guard cell photosynthesis/electron transport. Moreover, the finding that chlorophyll‐less stomata cause a ‘deflated’ thin‐shaped phenotype, suggests that photosynthesis in guard cells is critical for energization and guard cell turgor production.     

Model-based analysis of Arabidopsis leaf epidermal cells reveals distinct division and expansion patterns for pavement and guard cells

To efficiently capture sunlight for photosynthesis, leaves typically develop into a flat and thin structure. This development is driven by cell division and expansion, but the individual contribution of these processes is currently unknown, mainly because of the experimental difficulties to disentangle them in a developing organ, due to their tight interconnection. To circumvent this problem, we built a mathematic model that describes the possible division patterns and expansion rates for individual epidermal cells. This model was used to fit experimental data on cell numbers and sizes obtained over time intervals of 1 d throughout the development of the first leaf pair of Arabidopsis (Arabidopsis thaliana). The parameters were obtained by a derivative-free optimization method that minimizes the differences between the predicted and experimentally observed cell size distributions. The model allowed us to calculate probabilities for a cell to divide into guard or pavement cells, the maximum size at which it can divide, and its average cell division and expansion rates at each point during the leaf developmental process. Surprisingly, average cell cycle duration remained constant throughout leaf development, whereas no evidence for a maximum cell size threshold for cell division of pavement cells was found. Furthermore, the model predicted that neighboring cells of different sizes within the epidermis expand at distinctly different relative rates, which could be verified by direct observations. We conclude that cell division seems to occur independently from the status of cell expansion, whereas the cell cycle might act as a timer rather than as a size-regulated machinery.     

Regulation by amino acids of photorespiratory ammonia and glycolate release from ankistrodesmus in the presence of methionine sulfoximine

Methionine sulfoximine induced release of ammonia from illuminated cells of Ankistrodesmus braunii (Naegeli) Brunnth, in normal air, but less in air enriched to 3% CO₂. In normal air, methionine sulfoximine also induced glycolate release. Addition of either glutamate, glycine, or serine suppressed glycolate release, whereas glutamate and glycine at the same time stimulated ammonia release. The results indicate that inhibition of glutamine synthetase and thereby inhibition of photorespiratory nitrogen cycling restricts the sink capacity for glycolate in the photorespiratory carbon cycle. An external supply of glutamate, glycine, or serine seems to stimulate glyoxylate transamination and thus partly restores the sink capacity. Calculations of total glycolate formation rates in air from glycolate and ammonia release rates in the presence of methionine sulfoximine and glutamate revealed values of approximately 20 micromoles glycolate per milligram chlorophyll per hour on the average. Similar calculations led to an estimated rate of photorespiratory ammonia release in air, in the absence of methionine sulfoximine, of about 10 micromoles per milligram chlorophyll per hour on the average, a value comparable to the primary nitrogen assimilation rate of 8 micromoles per milligram chlorophyll per hour.     

An investigation of glycolate excretion in two species of blue-green algae

Summary The amount of ¹⁴C-glycolate excreted by Oscillatoria sp. and Anabaena flos-aquae is less than 1% of the ¹⁴C fixed by the algae during photosynthesis. Transfer of cells grown on 5% CO₂ in air to a medium of low bicarbonate concentration or treatment of the cells with isonicotinyl hydrazide (INH) during photosynthesis, caused little increase in glycolate excretion. -Hydroxysulfonates failed to stimulate massive excretion of glycolate. Although these blue-green algae excreted little glycolate, a significant proportion of the photosynthetically fixed carbon was excreted in the form of basic, neutral and acidic compounds, and such excretion was greater in 5% CO₂-grown cells than in air-grown cells.     

Cell division arrest by gamma-irradiation in photoautotrophic suspension culture of Euphorbia characias: Maintenance of photosynthetic capacity and overaccumulation of sucrose

Gamma-irradiation (250 Gy) applied to photoautotrophic cell suspensions of Euphorbia characias L. in the exponential growth phase led to the arrest of cell division and to a subsequent overaccumulation of sucrose and dry matter. From the fourth day of culture, the chlorophyll content and gross photosynthesis were not depressed by gamma-treatment nor by sugar accumulation. In both cultures, no difference was observed between oxygen uptake in the light at CO₂ saturating concentration and in the dark, suggesting that no change in energy-dissipative reactions took place after irradiation. A slight increase in oxygen uptake in both light and dark was observed in irradiated cells during the first four days. However, in the absence of limiting factors, the photosynthetic capacities of the dividing and irradiated non-dividing photoautotrophic cells were identical but higher than that of the non-dividing cells in the stationary growth phase. This suggests that gamma-irradiation arrests cell division by a mechanism different to that occuring in stationary-phase cultures. This may be of value in investigating the metabolism of secondary products.     

Impact of irradiance on the C allocation in the coastal marine diatom Skeletonema marinoi Sarno and Zingone

Elemental stoichiometry and organic composition were investigated in an Adriatic strain of Skeletonema marinoi, cultured at 25 [low light (LL)] and 250 [high light (HL)]µmol photon m^?2 s^?1. Inorganic carbon acquisition, fixation and allocation, and silicic acid and orthophosphate uptake were also studied. The C : P ratio was below the Redfield ratio, especially at LL. In HL cells, N quota was halved, C quota was similar, silica quota was lower, growth rate and long‐term net primary productivity were almost doubled, relative to LL cells. The HL : LL cell quota ratios were 6 for lipid, 0.5 for protein and 0.4 for carbohydrate. Phosphoenolpyruvate carboxylase (PEPc) and glutamine synthetase (GS) activities were unaffected by the growth irradiance; phosphoenolpyruvate carboxykinase (PEPck) was 2.5‐fold more active in LL cells. This suggests that in S. marinoi, C₄ photosynthesis is unlikely, PEPc is anaplerotic and PEPck may be involved in the conversion of lipid C to carbohydrates, especially in LL cells. Because about 50% of the cost for the production of an HL cell is caused by lipid biosynthesis, we propose that the preferential allocation of C to lipid at HL takes advantage of the relatively high volume‐based energy content of lipids, in an organism that reduces its size at each vegetative cell division.     

Influence of temperature and cell size on the division rate and chemical content of the diatom Chaetoceros curvisetum Cleve

Cell size and temperature influenced the division rate and chemical content of the diatom Chaetoceros curvisetum Cleve when grown at 15, 20 and 25°C in nutrient replete media. Cell-size dependent trends of division rate in individual clones changed with temperature in a complex fashion. Considerable interclonal variability in division rate within a restricted range of cell sizes was also found. Cellular levels of carbon, nitrogen, protein, chlorophyll a, and silicon were linearly related to cell size. Cellular levels of carbon and chlorophyll per unit volume and silicon per unit surface area changed with temperature. No temperature effect on cellular levels of nitrogen and protein was found.     

Studies on the biochemistry and fine structure of silica shell formation in diatoms

Summary Cell division in Navicula pelliculosa (Bréb.) Hilse, strain 668 was synchronized with an alternating regime of 5 h light and 7 h dark. Cell volume and dry weight increased only during the light period. DNA synthesis, which began during the third h of light, was followed sequentially by mitosis, cytokinesis, silicic acid uptake, cell wall formation, and cell separation. Silicification and a small amount of net synthesis of DNA, RNA and protein occurred during the dark at the expense of carbohydrate reserves accumulated during the light period. Cells kept in continuous light, after synchronization with the light-dark regime, remained synchronized through a second division cycle; the sequence of morphological events was the same as that in the light-dark division cycle, but the biosynthesis of macromolecular components changed from a stepwise to a linear pattern. The silicon-starvation synchrony was improved by depriving light-dark synchronized cells of silicic acid at the beginning of their division cycle, then resupplying silicic acid to cells blocked at wall formation.Abbreviation L light - D dark Portions based on a thesis submitted by W.M.D. to the University of California, San Diego in partial fulfillment of the requirements for the PH.D degree     

COMPARATIVE PHYSIOLOGICAL STUDY OF MARINE DIATOMS AND DINOFLAGELLATES IN RELATION TO IRRADIANCE AND CELL SIZE. II. RELATIONSHIP BETWEEN PHOTOSYNTHESIS,GROWTH, AND CARBON/CHLOROPHYLL a RATIO1,2

Photosynthetic rates, growth rates, cell carbon, cell protein, and chlorophyll a content of two diatom and two dinoflagellate species were measured. The microalgae were chosen to have one small and one large species from each phylogenetic group; the two size categories differed from each other by 1.5 orders of magnitude in terms of cell carbon or cell protein. The cultures for the experiments were grown under continuous light at an irradiance high enough for the light-saturation of growth for all four species. The four species were found to have similar maximum photosynthetic rates per unit chlorophyll a. The diatom species showed lower carbon/chlorophyll a ratios and higher photosynthetic rates per unit carbon than the dinoflagellates. The higher growth rates of the diatoms were shown to be related to their higher photosynthetic rates per unit carbon. The ecological significance of the physiological difference between these two groups of microalgae is discussed.