Effects of copper and zinc on growth,morphology, and metabolism of Asterionella japonica (Cleve) 1

When exposed to elevated levels of copper or zinc, the diatom Asterionella japonica (Cleve) showed a reduced cell division rate and a marked increase in cell size. Metal-treated cells had greater cell volumes, dry weights, carbon, nitrogen, chlorophyll, and DNA contents, all in approximately the same proportion as control cells. Two protoplasts often appeared to be contained within one frustule. Metal-treated cells photosynthesized at near-normal rates on a per chlorophyll basis and above normal rates on a per cell basis. Excretion of photosynthetically fixed carbon was depressed by metal treatment; 10–22% of fixed carbon was excreted in control cells and typically less than 1% in treated cells. Thus, metal-treated cells showed an uncoupling of photosynthesis from cell division and continued to enlarge when fixed carbon could not be excreted or utilized in cell division.Uptake of sulphate and silicic acid proceeded at slower rates than other processes (e.g., nitrogen uptake or photosynthesis) in copper-treated cells. Free amino acids in copper-treated cells totalled ≈ 10% of control cell levels, with greatest proportional declines in methionine, cysteine, aspartic acid, valine, and isoleucine. Copper-treated cells resuspended in fresh medium shrank to normal size when exposed to methionine (which they accumulated), although cell division rates did not return to normal. These cells excreted 2–3 times as much fixed carbon as comparable EDTA-treated or untreated cells, neither of which decreased in size. Copper-treated cells appeared indistinguishable from silicon-limited cells (i.e., cells not dividing for lack of silicon) in a copper-free medium. Cells treated with the sulfhydryl binder PCMB divided at reduced rates and also swelled in a manner comparable to copper-treated cells. The results suggest that toxic metals may bind to sulfhydryl groups on cell membranes, impairing normal membrane function and reducing silicic acid uptake and amino-acid synthesis, thereby resulting in depressed cell division rates.     

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.     

Environmental Control of Carbohydrate and Lipid Synthesis in Euglena

Dark-grown and light-grown nitrogen deficient Euglena accumulatedcarbohydrates and lipids under heterotrophic and phototrophicconditions. Although cellular lipid content increased, lipidas a percentage of dry weight was unaltered. Nitrogen deficientand sufficient heterotrophic cultures synthesized equal amountsof lipid per mole of exogenous carbon. Nitrogen deficiency doesnot alter the partitioning of available carbon between the synthesisof lipid and other cellular constituents but simply inhibitscell division so that the carbohydrate and lipid produced aredivided among fewer cells increasing the quantity of storageproducts per cell. Cellular chlorophyll content decreased onlywhen nitrogen deficient cells were maintained at high lightintensities suggesting that chlorophyll loss is due to a reductionin the rate of resynthesis of chlorophyll destroyed throughphotooxidative damage. Anaerobiosis triggered lipid synthesisand promoted carbohydrate breakdown in the dark and light. Evenafter six days of nitrogen deficiency, anaerobiosis triggeredadditional lipid accumulation and carbohydrate breakdown. Incontrast to nitrogen deficiency, anaerobiosis produced a preferentialincrease in the percentage of dry weight as lipid. This increasewas due as much to a decrease in cellular dry weight as it wasto the preferential utilization of available carbon for lipidsynthesis. ¹Present Address: The Mycology Center, Washington UniversitySchool of Medicine, St. Louis, MO 63178, U.S.A. ²Present Address: Department of Biology, University of Tampa,Tampa, Florida 33060, U.S.A. (Received September 16, 1987; Accepted January 18, 1988)     

Leaf Photosynthesis and Respiration of High CO(2)-Grown Tobacco Plants Selected for Survival under CO(2) Compensation Point Conditions

Four self-pollinated, doubled-haploid tobacco, (Nicotiana tabacum L.) lines (SP422, SP432, SP435, and SP451), selected as haploids by survival in a low CO₂ atmosphere, and the parental cv Wisconsin-38 were grown from seed in a growth room kept at high CO₂ levels (600-700 parts per million). The selected plants were much larger (especially SP422, SP432, and SP451) than Wisconsin-38 nine weeks after planting. The specific leaf dry weight and the carbon (but not nitrogen and sulfur) content per unit area were also higher in the selected plants. However, the chlorophyll, carotenoid, and alkaloid contents and the chlorophyll a/b ratio varied little. The net CO₂ assimilation rate per unit area measured in the growth room at high CO₂ was not higher in the selected plants. The CO₂ assimilation rate versus intercellular CO₂ curve and the CO₂ compensation point showed no substantial differences among the different lines, even though these plants were selected for survival under CO₂ compensation point conditions. Adult leaf respiration rates were similar when expressed per unit area but were lower in the selected lines when expressed per unit dry weight. Leaf respiration rates were negatively correlated with specific leaf dry weight and with the carbon content per unit area and were positively correlated with nitrogen and sulfur content of the dry matter. The alternative pathway was not involved in respiration in the dark in these leaves. The better carbon economy of tobacco lines selected for low CO₂ survival was not apparently related to an improvement of photosynthesis rate but could be related, at least partially, to a significantly reduced respiration (mainly cytochrome pathway) rate per unit carbon.     

Adaptation to quantum flux by the emerson photosynthetic unit

The size of the Emerson photosynthetic unit was measured in Chlorella pyrenoidosa strain no. 252 grown at light intensities between 50 and 1000 foot candles. The Emerson photosynthetic unit changed from a minimum size of 1970 molecules chlorophyll a + b/O₂ per flash in cells grown at 1000 foot candles to a maximum size of 3150 molecules chlorophyll a + b/O₂ per flash for cells grown at 50 foot candles. The size changes were interpreted as a partial adaptation where the trapping center antenna responded to changes in incident light intensity. Light-induced changes in chlorophyll content and size of the Emerson photosynthetic unit were directly related.     

The photosynthetic unit in chlorella measured by repetitive short flashes

Apparent size of the photosynthetic unit in Chlorella pyrenoidosa was estimated by the method of Emerson and Arnold: rate of oxygen evolution was measured under repetitive saturating flashes of about 10-microsecond duration separated by dark periods of 0.033 to 0.100 second. Cells used were taken from six steady state cultures maintained at different light intensities. Cell characteristics included a variation in chlorophyll content from 1 to 5%. Apparent size of the photosynthetic unit varied systematically with chlorophyll content in the range of 1560 to 2350 chlorophylls per O₂ per flash. Values for unit size showed no unusual statistical distribution and were not changed significantly by addition of low level background light at 645 or 705 nanometers. Maximal rate of unit turnover, calculated from light-saturated rate and unit size, varied inversely with chlorophyll content in the range of 70 to 180 per second.     

Photoinhibition and P700 in the Marine Diatom Amphora sp

The marine diatom Amphora sp. was grown at a light intensity of 7.0 × 10¹⁵ quanta centimeter⁻² second⁻¹. Light saturation of photosynthesis for these cells was between 6.0 and 7.0 × 10¹⁶ quanta centimeter⁻² second⁻¹. At light intensities greater than saturation, photosynthetic ¹⁴CO₂ fixation was depressed, while P700 unit size (chlorophyll a concentration/P700 activity) increased and number of P700 units per cell decreased. After a 1-hour exposure of Amphora sp. to a photoinhibitory light intensity of 2.45 × 10¹⁷ quanta centimeter⁻² second⁻¹, there was a 45 to 50% decrease in the rate of ¹⁴CO₂ fixation relative to the rate at the culture light intensity. There also was a 25% increase in P700 unit size and a 30% reduction in the number of P700 units per cell but no change in total chlorophyll a concentration. Following this period of photoinhibition, the cells were returned to a light regime similar to that in the original culture conditions. Within 1 hour, both number of P700 units per cell and P700 unit size returned to levels similar to those of cells which were kept at the culture light intensity. The rates of photosynthesis did not recover as rapidly, requiring 2 to 3 hours to return to the rate for the nonphotoinhibited cells. Our results indicate that a decrease in P700 activity (with a resultant increase in P700 unit size) may be partially responsible for the photoinhibition of algal photosynthetic carbon dioxide fixation.     

Labelling patterns of carbon-14 in net plankton during a winter-spring bloom

Weekly surface samples were collected in lower Narragansett Bay, Rhode Island, during the 1975 winter-spring bloom and fractionated by nets to nannoplankton (<20 μm) and total (< 158 μm) size fractions. Each size fraction was assayed for paniculate carbon, nitrogen, carbohydrate, protein, chlorophyll a, and cell counts. The <20 μm values were subtracted from the <158 μm values to estimate the composition of the 20 μm to 158 μm fraction (termed net plankton). As nutrients (primarily nitrogen) decreased to undetectable levels with the culmination of the diatom bloom, the ratios of protein/carbohydrate, carbohydrate/carbon, and carbon/chlorophyll a in the net plankton indicated the diatom population was increasingly nutrient-limited. Each size fraction was also incubated at a saturating light intensity with carbon-14; following filtration, the cells were extracted with solvents to obtain labelled polysaccharide and protein. The daily rates of polysaccharide and protein synthesis in the net plankton declined as the bloom entered the stationary phase. When the diatom population was at its maximum density the majority of the carbon-14 was found in the ethanol-soluble fraction; this may be due to high light intensities or nutrient effects.     

Photosynthetic Unit Size during the Synchronous Life Cycle of Scenedesmus

Apparent size of the photosynthetic unit (chlorophyll/O₂ per flash) was estimated by O₂ yield of repetitive short flashes on cell samples taken at various times from a synchronized culture (14-hour light, 10-hour dark) of Scenedesmus obliquus. Unit size was essentially invariant (< 10% variation) with a mean value of 1750 chlorophyll/O₂ per flash. In contrast, the light-saturated photosynthetic rate per unit chlorophyll, or turnover rate of the photosynthetic unit, varied with the life cycle, rising 40% in the first three hours of the light period and decaying slowly thereafter. The results are taken as evidence that the metabolic machinery is subject to far greater control and adjustment than is the photochemical machinery.     

Effects of environmental stresses on the cell cycle of two marine phytoplankton species

Cell cycle phase durations of cultures of Hymenomonas carterae Braarud and Fagerl, a coccolithophore, and Thalassiosira weissflogii Grun., a centric diatom, in temperature-, light- or nitrogen-limited balanced growth were determined using flow cytometry. Suboptimal temperature caused increases in the duration of all phases of the cell cycle (though not equally) in both species, and the increased generation time of nitrogen-limited cells of both species was due almost wholly to expansion of G₁ phase. In H. carterae light limitation caused only G₁ phase to expand, but in T. weissflogii both G₂ + M and G₁ were affected. These results are discussed in relation to cell division phasing patterns of these two species and to models of phytoplankton growth. Simultaneous measurements of protein and DNA on individual cells indicated that under all conditions, the protein content of cells in G₁ was a constant proportion of that of G₂ + M cells. Simultaneous measurements of RNA and protein on each cell indicated that the amounts of these two cell constituents were always tightly correlated. Under conditions of nitrogen limitation both protein and RNA per cell decreased to less than one-third of the levels found in nonlimited cells. This indicates, at least for nitrogen-replete cells, that neither protein nor RNA levels are likely to act as the trigger for cell cycle progression. Strict control by cell size is also unlikely since mean cell volume decreased as growth rates were limited by light and nitrogen supply, but increased with decreasing temperature.     

Adaptation of the Cyanobacterium Microcystis aeruginosa to Light Intensity

Light intensity adaptation (20 to 565 microeinsteins per square meter per second) of Microcystis aeruginosa (UV-027) was examined in turbidostat culture. Chlorophyll a and phycocyanin concentrations decreased with increasing light intensity while carotenoid, cellular carbon, and nitrogen contents did not vary. Variation in the number but not the size of photosynthetic units per cell, based on chlorophyll a/P₇₀₀ ratios, occurred on light intensity adaptation. Changes in the numbers of photosynthetic units partially dampened the effects of changes in light intensity on growth rates.     

NITROGEN LIMITATION INCREASES BREVETOXINS IN KARENIA BREVIS (DINOPHYCEAE): IMPLICATIONS FOR BLOOM TOXICITY1

Laboratory and field measurements of the toxin content in Karenia brevis cells vary by >4‐fold. These differences have been largely attributed to genotypic variations in toxin production among strains. We hypothesized that nutrient limitation of growth rate is equally or more important in controlling the toxicity of K. brevis, as has been documented for other toxic algae. To test this hypothesis, we measured cellular growth rate, chlorophyll a, cellular carbon and nitrogen, cell volume, and brevetoxins in four strains of K. brevis grown in nutrient‐replete and nitrogen (N)‐limited semi‐continuous cultures. N‐limitation resulted in reductions of chlorophyll a, growth rate, volume per cell and nirtogen:carbon (N:C) ratios as well as a two‐fold increase (1%–4% to 5%–9%) in the percentage of cellular carbon present as brevetoxins. The increase in cellular brevetoxin concentrations was consistent among genetically distinct strains. Normalizing brevetoxins to cellular volume instead of per cell eliminated much of the commonly reported toxin variability among strains. These results suggest that genetically linked differences in cellular volume may affect the toxin content of K. brevis cells as much or more than innate genotypic differences in cellular toxin content per unit of biomass. Our data suggest at least some of the >4‐fold difference in toxicity per cell reported from field studies can be explained by limitation by nitrogen or other nutrients and by differences in cell size. The observed increase in brevetoxins in nitrogen limited cells is consistent with the carbon:nutrient balance hypothesis for increases in toxins and other plant defenses under nutrient limitation.     

THE ACCLIMATED RESPONSE OF GROWTH,PHOTOSYNTHESIS, COMPOSITION,AND CARBON BALANCE TO TEMPERATURE IN THE PSYCHROPHILIC ICE DIATOM NITZSCHIA SERIATA1

Nitzschia seriata Cleve, a common member of marine bottom ice communities in the Arctic, was grown in unialgal batch cultures to test for compensatory mechanisms for the low temperatures (?1.8° C) typical of its natural habitat. The upper lethal limit for growth was between 12° and 15°C, and the optimum was between 6° and 12° C. The Arrhenius function adequately (R²= 73%) fitted the relationship between growth rate and temperature from – 1.6° up to 10° C, with an average Q₁₀ of 1.9 over the entire range. Light-saturated and light-limited rates of photosynthesis (normalized to chlorophyll a or cell carbon) showed complete compensation from 12° to 4° C. Photosynthetic rates, especially at light saturation, declined rapidly at temperatures below 4° C. Susceptibility to photoinhibition was greatest at the lowest growth temperatures. Cellular composition (chlorophyll a, protein, polysaccharide, and lipid contents) was not systematically related to temperature in any simple way, although cell size (carbon per cell) was maximal at the lowest growth temperature. Dark respiration was unmeasurably low (<0.015 day^?1) at all growth temperatures. The strategy of adaptation in N. seriata may be characterized as optimizing efficiency and compensation, rather than maximization, of growth rate.     

Variation of photosynthetic capacity with leaf age in an alpine orchid, Cypripedium flavum

Photosynthetic rate, chlorophyll fluorescence, leaf nitrogen and chlorophyll content of Cypripedium flavum were studied at different leaf ages. The photosynthetic capacity changed significantly with leaf age. Net photosynthesis and chlorophyll content peaked when leaf age was 60 days, decreasing at 30, 90 and 120 days. Stomatal conductance showed the highest value at 60 days, while mesophyll conductance decreased with increasing leaf age. Both leaf nitrogen content per unit area and leaf nitrogen content per unit mass decreased with increasing leaf age. The age-dependent variation in photosynthetic capacity could be linked to the changes in biochemical efficiency, leaf nitrogen content and CO₂ diffusion limitation.     

Culture conditions and the development of the photosynthetic mechanism; influence of light intensity on cellular characteristics of Chlorella

1. Chlorella pyrenoidosa has been grown in a continuous-culture apparatus under various light intensities provided by incandescent lamps, other conditions of culture being maintained constant. The harvested cells were analyzed for cell number, dry weight, nitrogen, and chlorophyll per unit cell volume. 2. Cell nitrogen and cell volume are parallel measures of cellular material over the range of light intensity studied. 3. The dry weight per cell volume increases slowly with light intensity, showing about a 20 per cent variation. 4. Chlorophyll concentration and cell number show a concomitant decrease with increasing light intensity, varying in such a way that there are always about the same number of chlorophyll molecules per cell. It is considered that this phenomenon has bearing on the interpretation of data which has led to the theory of the photosynthetic unit.     

On the Factors Which Determine Massive beta-Carotene Accumulation in the Halotolerant Alga Dunaliella bardawil

Dunaliella bardawil, a β-carotene-accumulating halotolerant alga, has been analyzed for the effect of various growth conditions on its pigment content, and compared with Dunaliella salina, a β-carotene nonaccumulating species. In D. bardawil, increasing light intensity and light period or inhibiting growth by various stress conditions such as nutrient deficiency or high salt concentration caused a decrease in the content of chlorophyll per cell and an increase in the amount of β-carotene per cell. As a result, the β-carotene-to-chlorophyll ratio increased from about 0.4 to 13 grams per gram and the alga changed its visual appearance from green to deep orange. D. salina grown similarly decreased in content of both chlorophyll and β-carotene per cell and the culture turned from green to yellowish. Low chlorophyll-containing cells of D. bardawil or D. salina exhibit very high photosynthetic rates when expressed on a chlorophyll basis (~600 micromoles O₂ evolved per milligram chlorophyll per hour).

Variation of pigment content in D. bardawil by a large variety of environmental agents has been correlated with the integral irradiance received by the algal culture during a division cycle. The higher the integral irradiance per division cycle, the lower the chlorophyll content per cell; the higher the β-carotene content per cell, and therefore the higher the β-carotene-to-chlorophyll ratio. The results are interpreted as indicating a protecting effect of β-carotene against injury by high irradiance under conditions of impairment in chlorophyll content per cell.

     

Regulation of the Photosynthesis Rhythm in Euglena gracilis: II. Involvement of Electron Flow through Both Photosystems

Rhythmic changes in the light reactions of Euglena gracilis have been found which help to explain the basic reactions effected in the circadian rhythm of O₂ evolution. Diurnal changes in the slope of light intensity plots indicated that the maximal rate of photosynthesis changed throughout the circadian cycle. No evidence was obtained consistent with the premise that changes in chlorophyll content, as measured by total chlorophyll or chlorophyll a/b ratio, or photosynthetic unit size are responsible for this rhythim.     

STUDIES ON THE AUTECOLOGY OF THE MARINE DIATOM THALASSIOSIRA NORDENSKIOELDII. II. THE INFLUENCE OF CELL SIZE ON GROWTH RATE,AND CARBON,NITROGEN, CHLOROPHYLL a AND SILICA CONTENT1

The effect of cell size on growth rates and some cellular contents of Thalassiosira nordenskioeldii Cleve has been measured at 0 and 10 C. At 0 C the growth rate did not vary with cell size. The 2 smallest clones at this temperature had reduced growth rates because of the induction of sexuality in that size range. The clones grown at 10 C showed a significant negative relationship between growth rate and valve diameter with the cell surface area/volume ratio positively related to growth rate. At both temperatures the smaller cells had proportionately more carbon and nitrogen/unit cell volume. The amount of chlorophyll a and silica/unit cell surface area increased with increasing cell surface area at both 0 and 10 C. Both the C/N and C/chl a ratios showed no significant change with cell size at either temperature but there was a significant increase in the C/chl a ratio at 0 C. The C/Si ratio decreased with increasing cell size at both 0 and 10 C.     

Effects of Flashes of Red or Blue Light on the Composition of Starved Chlorella pyrenoidosa

Autotrophically grown cells of Chlorella pyrenoidosa (211-8b) were starved 3 to 4 days in darkness, flashes of blue light, or flashes of red light. The blue flashes were sufficient to maintain the maximal rate of light-stimulated oxygen uptake during short term experiments. However, after 24 hours, the respiration rate in red flashes was equal to, or greater than, the rate in blue flashes. Starvation in darkness reduced the chlorophyll content by 11%, altered the blue absorbance of the nonsaponifiable material only 1 to 2%, and reduced the dry weight by 13%. Starvation in the presence of blue or red flashes reduced the dry weight by an additional 11 or 12% respectively. Protein per unit cell volume was not changed significantly during 3 to 4 days starvation in darkness or in blue flashes, even though dry weight per unit cell volume decreased 13% in darkness and 23% in blue flashes. In contrast, cells starved under red flashes showed a 20% decrease in protein per unit cell volume and a 24% decrease in dry weight per unit cell volume.     

Effects of Nitrate Application on Amaranthus powellii Wats. : I. Changes in Photosynthesis, Growth Rates, and Leaf Area

Physiological effects of different nitrate applications were studied using the C₄ plant, Amaranthus powellii Wats. Plants were grown in a controlled environment chamber and watered daily with nutrient solutions containing 45, 10, 5, or 1 millimolar nitrate. Chloride and sulfate were used to keep the cation and phosphate concentrations constant. Total leaf nitrogen concentration, chlorophyll concentration, specific leaf mass, leaf area, relative growth rate, relative leaf growth rate, unit leaf rate (increase of dry mass per unit leaf area per day), net photosynthetic rate, and incident quantum yield decreased with decreasing nitrate concentration. The per cent decrease of unit leaf rate was similar to the decrease of light-saturated net photosynthetic rate; however, the decrease in relative growth rate was less than that of unit leaf rate because leaf area ratio (leaf area per unit dry mass) increased with decreasing nitrate concentration. Essential mineral concentrations per unit leaf area were about equal among all treatments. Leaf expansion, determined by stomatal density, decreased except for the 1 millimolar treatment which showed relatively more cell expansion but less cell division. Decreased nitrate application was correlated with higher osmotic potentials and lower pressure potentials (determined by pressure-volume curves), whereas leaf water potentials were equal among treatments. Even though total leaf area and shoot mass decreased with decreasing applied nitrate, the increase of the leaf area ratio may be related to selection for the highest possible growth rate.