Culture conditions and the development of the photosynthetic mechanism; influence of light intensity on photosynthetic 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. Light intensity curves for cells immersed in the No. 11 Warburg buffer and in Knop''s solution + 4.4 per cent CO₂ at a saturating light intensity were determined as characteristics of the photosynthetic mechanism. These characteristics were referred to the centrifuged cell volume as an index of quantity of cellular material. 2. Cells grown at intensities in the range of about 35 f.-c. develop a capacity for a high rate of photosynthesis (c.mm. O₂/hour/c.mm. cells). At culture intensities above or below this range the cells produced have a lower capacity for photosynthesis. A similar effect is observed for rate of photosynthesis per unit dry weight or rate per unit cell nitrogen. 3. The rate of photosynthesis per cell or rate per unit chlorophyll shows no maximum at any light intensity of culture but increases continuously throughout the range of light intensities studied. 4. Maximum rate of growth is attained at a light intensity of about 100 f.-c. The hypothesis is advanced that at culture intensities above that needed to give maximum rate of growth (100 f.-c.) a mechanism is developed which opposes the photosynthetic process and removes the photosynthetic products. 5. The low capacity for photosynthesis shown by cells grown at culture intensities below 35 f.-c. finds no immediate explanation. 6. The shape of the light intensity curve is markedly affected by the light intensity at which the cells have been cultured. Cells grown at lower intensities give light intensity curves approaching the Blackman type with a short transitional region between light limitation and light saturation.     

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.     

毛白杜鹃‘紫萼’的光适应性及最适光强的研究

从形态、生理角度研究了杭州园林中应用最广泛的杜鹃‘紫萼’(Rhododendron mucronatum cv Plenum)的光适应性和最适光强生境。结果表明:随着叶片遮荫程度的增加,杜鹃的叶面积和叶绿素含量增加;光补偿点、光饱和点及暗呼吸强度下降,说明杜鹃对弱光生境有一定的适应性。另一方面,随着相对光强的增加,叶片厚度,比叶重以及栅栏组织、海绵组织厚度及其比值,可溶性蛋白质及净光合速率增加,表现出对阳生生境更好的适应性。在生境65％全光照时,植株在形态,解剖及生理上均处于最佳状态。因此,65％全光照的生境是毛鹃‘紫萼’的最佳光生境。     

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.     

Effects of growth irradiance levels on the ratio of reaction centers in two species of marine phytoplankton

Cells of two species of single-celled marine algae, the diatom Skeletonema costatum (Greve), Cleve, and the chlorophyte Dunaliella tertiolecta Butcher, were cultured in white light of high (500-600 microeinsteins per square meter per second) and low (30 microeinsteins per square meter per second) intensity. For both algal species, cells grown at low light levels contained more chlorophyll a and had a lower ratio of chlorophyll a to chlorophylls b or c than did cells grown at high light levels. When photosynthetic unit sizes were measured on the basis of either oxygen flash yields or P₇₀₀ photooxidation, different results were obtained with the different species. In the chlorophyte, the cellular content of photosystem I (PSI) and photosystem II (PSII) reaction centers increased in tandem as chlorophyll a content increased so that photosynthetic unit sizes changed only slightly and the ratio PSI:PSII reaction centers remained constant at about 1.1. In the diatom, as the chlorophyll content of the cells increased, the number of PSI reaction centers decreased and the number of PSII reaction centers increased so that the ratio of PSI:PSII reaction centers decreased from about unity to 0.44. In neither organism did photosynthetic capacity correlate with changes in cellular content of PSI or PSII reaction centers. The results are discussed in relationship to the physical and biological significance of the photosynthetic unit concept.     

The effects of light and nitrogen on photosynthesis,leaf characteristics,and dry matter allocation in the chaparral shrub,Diplacus aurantiacus

Summary Plants of Diplacus aurantiacus, a successional shrub common in California chaparral, were grown under controlled conditions in which either quantum flux density or nitrogen availability was varied. Photosynthesis and leaf nitrogen content were determined on a leaf area and a leaf weight basis, and whole plant growth was monitored.There was a direct relationship between photosynthesis and leaf nitrogen content on both area and weight bases. Reduced light intensity of the growth environment resulted in reductions in light-saturated photosynthesis and nitrogen content on an area basis, but not on a weight basis. With reduced nitrogen availability, photosynthesis and leaf nitrogen content per unit leaf weight decreased.Resource use efficiency increased as the resource became more limiting. The results are consistent with a model of plant growth in which net carbon gain of the leaf is maximized. Abbreviations. For brevity, the following set of abbreviations is used in presenting and discussing the results. P/area and N/area are, respectively, photosynthesis and leaf nitrogen content per unit leaf area. P/wt and N/wt are the same quantities per unit leaf dry weight. SLW (specific leaf weight) is dry weight per unit leaf area. RGR (relative growth rate) is the relative rate of increase in shoot dry matter per day     

Effects of illumination and nitrogen starvation on accumulation of arachidonic acid by the microalga Parietochloris incisa

Parietochloris incisa is a unicellular freshwater green alga capable of accumulating high amounts of the valuable long-chain polyunsaturated arachidonic acid (AA) in triacylglycerols (TAG) of cytoplasmic oil bodies. To find the cultivation conditions providing maximum AA yield, the effects of illumination and N-availability on the dry weight (DW), chlorophyll, carotenoid, and AA content were studied. Under nitrogen starvation, TAG accounted for over 30% of dry weight (DW) and the AA content became as high as about 55% of total fatty acids. For biomass accumulation, light intensity of ca 400 μE m^?2 s^?1 was found to be optimal for growing P. incisa on a complete medium. Lower light intensities (or a higher cell density of inoculum) resulted in a higher AA yield when the alga was cultivated on nitrogen-free media. In the absence of nitrogen, algal cells were unable to cope with high illumination and suffered from photooxidative damage, whereas the nutrientsufficient culture survived under such illumination conditions, probably due to accumulation of carotenoids. Nitrogen-deprived P. incisa cells displayed elevated sensitivity to light.     

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.     

The Effect of Nitrogen on the Growth and Sugar Content of Sugar-beet

Nitrogen fertilizer applied to sugar-beet increased plant androot dry weight and leaf area, and decreased the sugar contentof the roots per cent of both fresh and dry weight. Change inleaf area accounted wholly for the increase in plant dry weightproduced by nitrogen, because net assimilation rate was unaffected.Nitrogen did not alter the partition of the total assimilatebetween roots and shoots, but increased the fraction of totalassimilate entering the roots that was used in growth, at theexpense of that stored as sugar. Thus, plants with more nitrogenhad a smaller proportion of their root dry weight as sugar becausemore was metabolized in growth of the roots, and not becauseless entered the roots. The heavier roots of plants given more nitrogen were largerin cross-sectional area because the areas of both parenchymaand vascular zones of each peripheral ring within the root werelarger; the number of rings was not increased. Nitrogen increasedthe areas of the tissues in these zones by enlarging cell volumes,not by increasing the number of cells within the tissues. Increasein cell volume was accompanied by proportional increases inthe weights of non-sugar dry matter per cell and water per cell,but the amount of sugar per cell was proportional to cell volumeonly during the initial stage of cell expansion up to cell volumesof about 15x10^–8 cm²; thereafter it was less than proportional,so that sugar per cent of both fresh and dry weight decreasedas cell size increased beyond 15x10^–8 cm². The relationof sugar per cell to cell volume was the same with both amountsof nitrogen given. This implies that increase in nitrogen supplymade the sugar concentration of the root less by increasingthe size of the root cells and not by a specific effect on sugarstorage.     

Predicting Production in Light-Limited Continuous Cultures of Algae

Equations relating productivity, growth rate, cell concentration, and light absorption lead to the prediction that, when incident light is below saturating intensity, maximal productivity will occur at half the maximal growth rate. The freshwater alga Chlorella pyrenoidosa TX71105 and the marine alga Dunaliella tertiolecta were grown in a small continuous culture apparatus with turbidostatic control. With both cultures, the cell concentration showed a linear decrease with dilution rate. Productivity was maximal at about one-half the maximal dilution rate. Average mass per cell increased near the maximal dilution rate, causing some asymmetry in the productivity versus dilution rate curve. The chlorophyll content per unit mass decreased in this region, but the chlorophyll content per cell remained constant. Best production rate in a light-limited algal culture was obtained when the growth rate at very low cell concentration was determined in the apparatus and the dilution rate was set at one-half that value.     

Chlorophyll synthesis in chlorella I. Occurrence of a lag phase on initiation of a dilute culture

A culture of Chlorella established by 30-fold dilution of a culture already grown to a level of 15 ml packed cell volume per liter produces little chlorophyll for approximately 12 hours. Investigation of other characteristics such as nitrogen incorporation, increase in packed cell volume and dry weight as well as RNA level show all of these to increase without any significant lag. α-Linolenate, which can be considered as a chloroplast marker, increased markedly. Photosynthetic oxygen evolution and respiration as well as the heme enzyme, catalase, increase also, indicating that the lag in chlorophyll synthesis is not due to a general inability to produce the porphyrin moiety.     

Studies on the Production of Digitalis Cardenolides by Plant Tissue Culture: II. EFFECT OF LIGHT AND PLANT GROWTH SUBSTANCES ON DIGITOXIN FORMATION BY UNDIFFERENTIATED CELLS AND SHOOT-FORMING CULTURES OF DIGITALIS PURPUREA L. GROWN IN LIQUID MEDIA

Undifferentiated, highly chlorophyllous cell cultures; undifferentiated white cell cultures; green, shoot-forming cultures; and white, shoot-forming cultures of Digitalis purpurea L. were established and subcultured every 3 weeks in liquid media in the light or in the dark. The digitoxin content, the chlorophyll content, and the ribulose bisphosphate carboxylase activity of these cultures were assayed. The light-grown, green, shoot-forming cultures accumulated considerable amounts of digitoxin (about 20 to 40 micrograms per gram dry weight), and the white, shoot-forming cultures without chloroplasts accumulated about one-third that amount of digitoxin. The chlorophyll content and the ribulose bisphosphate carboxylase activity of the undifferentiated green cells were about the same as they were in the green, shoot-forming cultures, but the digitoxin content of the former was extremely low (about 0.05 to 0.2 microgram per gram dry weight), which is about the same as that in undifferentiated white cells without chloroplasts. Thus, it was concluded that the chloroplasts are not essential for the synthesis of digitoxin in Digitalis cells. The optimum concentrations of the tested compounds for accumulation of digitoxin were: benzyladenine, 0.01 to 1 milligram per liter; indoleacetic acid, 0.1 to 1 milligram per liter; α-naphthaleneacetic acid; 0.1 milligram per liter; and 2,4-dichlorophenoxyacetic acid, 0.01 milligram per liter.     

Photosynthetic activity in synchronized cultures of algae and its dependence on light intensity

Summary Photosynthetic capacity at several levels of illuminance was investigated in cells of the successive developmental stages of the high temperature strain, Chlorella 7-11-05. The saturating light intensity, rates of photosynthesis at light saturation and at half-saturation, and the slope of the light dependent portion of the light intensity curves were proved to be bound with the developmental status of cells. The effect of the choosing of the basis for calculations of photosynthetic activity — whether packed cell volume, dry weight, nitrogen, or chlorophyll content—was discussed. The fact was stressed that with the available synchronization technique the observable amplitude of changes in metabolic activity in the course of cell development is a minimal one and the actual fluctuations in photosynthetic rates in non-synchronized suspensions in the course of the life cycle of the individual cells are expected to be higher than those recorded for synchronized populations.     

Stimulation of Nodulation in the Clover-Rhizobium trifolii 0403 System by Penicillin and Mecillinam

The number of nodules produced per clover seedling inoculated with Rhizobium trifolii 0403 can be increased almost 2-fold by the addition of penicillin or mecillinam. Two-day-old dutch white clover seedlings grown in 250 milliliter boston round jars containing agar-solidified plant growth medium were inoculated with exponentially growing Rhizobium trifolii 0403 cells. Penicillin or mecillinam (100 micrograms per milliliter) were added immediately or after 24 hours. Following 42 days growth, 10 replicate sets of 5 plants for each treatment were assayed for nodule number, plant dry weight, and Kjeldahl nitrogen. Both antibiotics increased nodule number, plant dry weight, and Kjeldahl nitrogen. Increases in nodule number and dry weight were statistically significant. The range of values in Kjeldahl nitrogen was so extensive as to make the data insignificant at the P < 0.05 level, however nodule number, plant dry weight, and Kjeldahl nitrogen displayed a significant correlation with each other. There were no significant differences in treatment with either antibiotic or with time of treatment. Nodule number increased by about 85%, and plant dry weight and nitrogen increased by about 30%.     

Effect of Light Quality on the Composition, Function, and Structure of Photosynthetic Thylakoid Membranes of Asplenium australasicum (Sm.) Hook

The effect of light quality on the composition, function and structure of the thylakoid membranes, as well as on the photosynthetic rates of intact fronds from Asplenium australasicum, a shade plant, grown in blue, white, or red light of equal intensity (50 microeinsteins per square meter per second) was investigated. When compared with those isolated from plants grown in white and blue light, thylakoids from plants grown in red light have higher chlorophyll a/chlorophyll b ratios and lower amounts of light-harvesting chlorophyll a/b-protein complexes than those grown in blue light. On a chlorophyll basis, there were higher levels of PSII reaction centers, cytochrome f and coupling factor activity in thylakoids from red light-grown ferns, but lower levels of PSI reaction centers and plastoquinone. The red light-grown ferns had a higher PSII/PSI reaction center ratio of 4.1 compared to 2.1 in blue light-grown ferns, and a larger apparent PSI unit size and a lower PSII unit size. The CO₂ assimilation rates in fronds from red light-grown ferns were lower on a unit area or fresh weight basis, but higher on a chlorophyll basis, reflecting the higher levels of electron carriers and electron transport in the thylakoids.

The structure of thylakoids isolated from plants grown under the three light treatments was similar, with no significant differences in the number of thylakoids per granal stack or the ratio of appressed membrane length/nonappressed membrane length. The large freeze-fracture particles had the same size in the red-, blue-, and white-grown ferns, but there were some differences in their density. Light quality is an important factor in the regulation of the composition and function of thylakoid membranes, but the effects depend upon the plant species.

     

Phytochrome Effects on the Relationship between Chlorophyll and Steady-State Levels of Thylakoid Polypeptides in Light-Grown Tobacco

The effects of phytochrome status on chlorophyll content and on steady-state levels of thylakoid proteins were investigated in green leaves of Nicotiana tabacum L. plants grown under white light. Far-red light given either as a pulse at the end of each photoperiod, or as a supplement to white light during the photoperiod, reduced chlorophyll content per unit area and per unit dry weight. These differences were also observed after resolving chlorophyll-containing polypeptides by gel electrophoresis. Chlorophyll a:b ratio was unchanged. Both Coomassie blue-stained gels and immunochemical analyses showed that, in contrast to the observations in etiolated barley (K Apel, K Kloppstech [1980] Planta 150: 426-430) and pea (J Bennett [1981] Eur J Biochem 118: 61-70) seedlings, and in etiolated tobacco leaves (this report), in fully deetiolated tobacco plants changes in chlorophyll content were not correlated with obvious changes in the steady-state levels of thylakoid proteins (e.g. light-harvesting, chlorophyll a/b-binding proteins).     

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.     

C and N utilization of two lettuce genotypes during growth under non-varying light conditions and after changing the light intensity

Growth and N-incorporation in two lettuce genotypes ( Lactuca sativa L. cv. Deci minor and cv. Grosse brune), which differ significantly in nitrate accumulation, were studied. Under constant environmental conditions cv. Deci minor produced more fresh and dry weight than cv. Grosse brune. Cultivar Deci minor also produced more fresh weight per mmol N absorbed than cv. Grosse brune, and contained less organic nitrogen in the dry matter, but accumulated more nitrate. As cv. Deci minor showed a higher fresh and dry weight production per mmol N absorbed than cv. Grosse brune, it used its nitrogen more efficiently.
When the light intensity was decreased, the growth of both cultivars decreased, and the fresh weight production per mmol N absorbed increased. After reduction of the light intensity, cv. Deci minor maintained a higher fresh weight production per N absorbed than before, whereas cv. Grosse brune returned to its original level. After decrease of the light intensity, an increased nitrate concentration in the cell sap was accompanied by a decreased concentration of organic compounds in both cultivars. The organic nitrogen level in the dry matter remained constant after the higher intensity was reduced. However, due to the decreased dry weight percentage, the demand for nitrogen for protein synthesis decreased on fresh weight basis.
It can be concluded that the two cultivars differ in their partition of C and N between dry matter and cell sap. Nitrate accumulation in preference to accumulation of organic compounds does not automatically result from a shortage of organic compounds. The high accumulation of nitrate of cv. Deci minor enables it to use more carbohydrates for structural growth than cv. Grosse brune.     

Functional organization and plasticity of the photosynthetic unit of the cyanobacterium Anacystis nidulans

Anacystis nidulans grown under high and low light, 100 and 10 μE m^?2 s^?1, respectively, was analyzed with respect to chlorophyll/P700, phycobiliproteins/P700, chlorophyll/cell, and oxygen evolution parameters. The photosynthetic unit sizes of this cyanobacterium, measured as the ratio of total chromophores (chlorophyll and bilin) to P700, were shown to be similar to those of higher plants and green algae. High light grown cells possessed a photosynthetic unit consisting of a core of 157 ± 6 chlorophyll a molecules per P700 associated with a light harvesting system of 95 ± 3.5 biliprotein chromophores. Low light grown cells had substantially more biliprotein chromophores per P700 (125 ± 3.1) than high light cells, but showed no significant difference in the numbers of chlorophyll a molecules per P700 (149 ± 4). Analyses of aqueous biliprotein extracts indicate that low light grown cells produce proportionately more phycocyanin relative to allophycocyanin than high light cells. Calculations of the molecular weight of biliproteins per P700 suggest that there is less than one phycobilisome per reaction center I under both growth conditions. Differences in chlorophyll/cell ratios and oxygen evolution characteristics were also observed. High light cells contain 6.3 × 10^?12 mg chlorophyll cell^?1, while low light grown cells contain 12.8 × 10^?12 mg chlorophyll cell^?1. Photosynthetic oxygen evolution rate vs. light intensity curves indicate that high light grown cells reach maximal levels of oxygen evolution at higher light intensity than low light grown cells. Maximal rates of oxygen evolution were 16.6 μmol oxygen min^?1 (mg chlorophyll)^?1 for high and 8.4 μmol oxygen min^?1 (mg chlorophyll)^?1 for low light cells. Maximal oxygen evolution rates per cell were equivalent for both cell types, although the amount of P700 per cell was lower in high light cells. High light grown cells are therefore capable of producing more oxygen per reaction center I than low light grown cells.