COMPARISON OF GROWTH AND SINKING RATES OF NON-COCCOLITH- AND COCCOLITH-FORMING STRAINS OF EMILIANIA HUXLEYI(PRYMNESIOPHYCEAE) GROWN UNDER DIFFERENT IRRADIANCES AND NITROGEN SOURCES1

We examined the effect of the presence or absence of coccoliths on the growth and sinking rates of an oceanic isolate of the coccolithophore Emiliania huxleyi (Lohmann) Hay et Mohler isolated from the northeastern subarctic Pacific. Coccolith-forming and non-coccolith-forming (i.e. naked, nonmotile) strains were obtained from the same isolate and grown under both saturating and limiting irradiance levels with either nitrate or ammonium as the primary nitrogen source. Sinking rate, growth rate, and cell volume (excluding coccoliths) were measured for each culture. Under saturating irradiance, coccolith-forming cells grew significantly slower than naked cells, had significantly higher sinking rates, and had larger cell volumes than naked cells. Under limiting irradiance levels, growth rates of the two strains were identical, sinking rates were higher for coccolith-forming cells in stationary-phase cultures only, and cell volumes remained greater for coccolith-forming cells. The sinking rates achieved for this ubiquitous coccolithophore ranged from <0.1 to 0.5 m · d^?1. Sinking rates were not statistically different between coccolith-forming and naked strains of E. huxleyi under limiting irradiance conditions for log-phase cultures, but sinking rates were greater for coccolith-forming cells under some of the other conditions tested. However, the average sinking rate was never more than twice as great as for coccolith-forming cells, with the exception of nitrate-grown, senescent cells under limiting irradiance (3.4 times greater). Cell volumes (excluding coccoliths) were consistently ca. 1.5 times greater for coccolith-forming cells than for naked cells. Nitrogen source had an effect on growth rate and cell volume, with ammonium-grown cultures growing faster and having larger cell volumes than nitrate-grown cultures of both strains. However, despite the difference in growth rate and cell volume, nitrogen source had little if any effect on sinking rate.     

Effect of Ammonia and Nitrate on Growth, Photosynthesis, and Ribulosediphosphate Carboxylase Content of Dunaliella tertiolecta

When the marine Chlorophycean flagellate Dunaliella tertiolecta Butcher was grown with short photoperiods of bright light, the use of ammonia rather than nitrate as a nitrogen source led to a 30 % reduction of the doubling time of cell matter. The cell cycle (onset of light to completion of cell division) was shortened by about 10% only. Ammonia-grown cells possessed a greater capacity for photosynthetic oxygen evolution at light saturation than did nitrate-grown cells; their content of ribulosediphosphate carboxylase was likewise greater. The faster growth of Dunaliella tertiolecta with ammonia may be partly a consequence of a general increase in net protein synthesis resulting in a greater content of photosynthetic enzymes.     

Far-red light inhibits growth but promotes carotenoid accumulation in the green microalga Dunaliella bardawil

It has been demonstrated that far-red light reduces growth of marine phytoplankton and that light quality controls growth and photosynthetic metabolism in algae. The green halotolerant microalga, Dunaliella bardawil, accumulates high amounts of β-carotene (up to 10% of its dry weight) under conditions of high light or nutrient limitation. The influence of increasing irradiance and of far-red light in D. bardawil was studied. Continuous irradiance was provided by white fluorescent lamps alone (WL) or supplemented with far-red Linestra lamps (WL+FR). For both types of light, cultures were acclimatized at increasing irradiances (50-300 µmol m^?2 s^?1), and cell density, photosynthetic activity and pigment content were determined. Cell density increased with the photon irradiance, and was higher in WL than in WL+FR under the same irradiance, but the reverse occurred in respect of cell volume. Growth rate was higher under WL+FR. Far-red light induced faster growth but reduced the maximal cell density of the cultures. Chlorophyll a concentration was higher in white light, but total carotenoid content increased dramatically in both far-red light treatments (about 50% on a per cell basis) and with the increase of irradiance. Our results show that far-red light has a significant influence on growth and photosynthesis of D. bardawil, inducing a decrease in cell density, photosynthetic activity and chlorophyll concentration, and an increase in growth rate, cell volume and carotenoid content.     

CHARACTERIZATION OF TWO CHLORELLA VULGARIS (CHLOROPHYCEAE) STRAINS ISOLATED FROM WASTEWATER OXIDATION PONDS1

We report here on the characterization and isolation of two ecotypes of Chlorella vulgaris Beyerinck that coexist in wastewater reservoirs. One ecotype (C1) contains high amounts of chlorophyll b, is capable of autotrophic growth, and can utilize only a few organic solutes for growth. The second ecotype (C2) contains low amounts of chlorophyll b, requires vitamin B₁₂, and can support its growth with a broad range of organic compounds. Of the two ecotypes, the latter showed slower growth rates when light was the sole source of energy. Cells of C2-type Chlorella attained higher photosynthetic activities than C1-type cells at saturating irradiances. However, their low chlorophyll b content and lower light utilization efficiency suggest that C2-type Chlorella contains relatively low amounts of light-harvesting antennae, a disadvantage in severely light-limited ecosystems like wastewater reservoirs. We hypothesize that the two Chlorella types coexist by adopting different lifestyles: C1-type cells rely largely on their photosynthetic potential for energy conservation and growth, whereas C2-type cells may exploit their heterotrophic properties for this purpose.     

INFLUENCE OF IRRADIANCE ON CELL VOLUME AND CARBON QUOTA FOR TEN SPECIES OF MARINE PHYTOPLANKTON1

Ten species of marine phytoplankton were grown under a range of photosynthetic photon flux densities (PFDs) and examined for variation in cell volume and carbon quota. Results suggest that in response to low PFDs phytoplankton generally reduce their cell volume and frequently reduce their carbon quota. A significant linear relationship between the log of PFD (I) and cell volume (in nine of ten species) and log I and carbon quota (four of ten species) was demonstrated. When exposed, to a transient in light intensity, Thalassiosira pseudonana (Hustedt, clone 3H) Hasle and Heimdal underwent a rapid adaptation in cell volume and carbon quota. Cells going from low light to high light reached maximum mean cell volume within 5 h, and cells going from high light to low light reached a minimum mean cell volume within 12 h. The resulting kinetic constant (k; a measure of the rate of adaptation) was considerably larger than previously reported k values. Ditylum brightwellii (West) Grunow increased in length but did not increase in width during a transient to increased irradiance. Nutrient limitation was shown to override PFD in determining cell volume and carbon quota for Heterosigma akashiwo Hada. Cells grown at equivalent irradiances but N-limited, were smaller than light-limited and nutrient-saturated cells. Therefore, cell volume and carbon quota do not have the same relationship with PFD when factors other than PFD control growth rate. The ecological implications of reduced cell volumes and carbon quotas with decreasing PFD include possible impacts on CO₂ budgets, an influence on sinking rates, potential changes in predation rates, and surface area/cell volume benefits.     

不同光照条件下喜阴植物谢君魔芋对稳态和动态光源的响应特征

谢君魔芋(Amorphophallus xiei)是起源于云南西南地区热带雨林的典型喜阴植物,近年来得到了广泛种植和推广,在种植过程中,谢君魔芋需要采用遮荫栽培模式。为了揭示谢君魔芋对光照强度的适应策略,该研究探讨了生长在不同光照强度下(透光率为50%、29%、17%、7%)谢君魔芋叶片的光合作用特征、光合诱导特征、光合色素含量以及叶片氮素(N)含量和N分配。结果表明:随着生长环境光照强度的降低,单位叶面积和单位叶质量最大净光合速率、光合色素含量、最大羧化速率、最大电子传递速率及比叶面积均增大,而暗呼吸和光补偿点均减小。在光合诱导过程中,生长在透光率为17%光环境中的谢君魔芋完成50%光合诱导所需的时间最短,约为81.4 s;在光诱导进行10 min时,诱导状态最高,为87.3%。完成50%和90%光合诱导所需的时间与低光下初始气孔导度呈负相关关系。随着生长光照强度降低,叶片中的N分配到羧化组分和生物能转化组分中的比例先增大后减小,在透光率为17%的光环境下具有最大值;而叶片中的N分配到捕光色素组分中的比例随着生长环境光照强度降低而增加。该研究结果表明,喜阴植物谢君魔芋通过加强对低光和动态光源的利用能力及有效的N资源分配策略来适应低光照环境。     

INTERACTIVE EFFECTS OF IRRADIANCE AND TEMPERATURE ON THE PHOTOSYNTHETIC PHYSIOLOGY OF THE PENNATE DIATOM PSEUDO‐NITZSCHIA GRANII (BACILLARIOPHYCEAE) FROM THE NORTHEAST SUBARCTIC PACIFIC1

This study examined how light and temperature interact to influence growth rates, chl a, and photosynthetic efficiency of the oceanic pennate diatom Pseudo‐nitzschia granii Hasle, isolated from the northeast subarctic Pacific. Growth rates were modulated by both light and temperature, although for each irradiance tested, the growth rate was always the greatest at ～14°C. Chl a per cell was affected primarily by temperature, except at the maximum chl a per cell (at 10°C) where the effects of light were noticeable. At both ends of the temperature gradient, cells displayed evidence of chlorosis even at low light intensities. Chl fluorescence data suggested that cells at 8°C were significantly more efficient in their photosynthetic processes than cells at 20°C, despite having comparable concentrations of chl. Cells at low temperature showed photosynthetic characteristics similar to high‐irradiance‐adapted cells. The decline of growth rates beyond the optimum growth temperature coincided with the cell's inability to accumulate chl in response to increasing temperature. The decline in photosynthetic ability at 20°C was likely due to a combination of high‐temperature stress on cellular membranes and a decline in chl. Our results highlight the important interactions between light and temperature and the need to incorporate these interactions into the development of phytoplankton models for the subarctic Pacific.     

Physiological effects of the presence and absence of gas vacuoles in the blue-green alga,Microcystis aeruginosa Kuetz. emend. Elenkin

Physiological evidence was obtained for a light shielding role for gas vacuoles inMicrocystis aeruginosa Kuetz. emend. Elenkin, by comparing photosynthetic oxygen evolution, growth behaviour and pigment composition of cells with intact or collapsed gas vacuoles. The oxygen evolution rates were strongly dependent on cell concentration, a maximum rate for cells with intact gas vacuoles occurring at about 1.4×10⁹ cells/ml and for cells with collapsed gas vacuoles at about 2.5×10⁹ cells/ml. By using light saturation curves for oxygen evolution, it was estimated that at low light intensities up to 30% of the photosynthetically useable light was shielded at a cell concentration of 6×10⁸ cells/ml. Collapsing the gas vacuoles twice daily did not alter the initial growth rate of the cultures, but enabled them to reach a higher final cell density. Collapsing of gas vacuoles during growth for about four generations resulted in a lower level of all acetone soluble pigments with a greater relative reduction in carotenoids than in chlorophyll a. Collapse of the gas vacuoles does not alter the cell volume. Various optical interactions which could account for light shielding are discussed.     

NaCl-induced photoinhibition and recovery of the photosynthetic activity of a <Emphasis Type="Italic">katG</Emphasis><Superscript>−</Superscript> mutant of cyanobacterium <Emphasis Type="Italic">Synechocystis</Emphasis> sp. PCC 6803

The joint effects of 0.5 M NaCl and light of different intensities on the activity of the photosynthetic apparatus and ATP content in cells of the katG ⁻ mutant of cyanobacterium Synechocystis sp. PCC 6803 have been studied. The mutant demonstrated a higher photoinhibition rate and a slower rate of recovery compared with the wild type, as shown by measurements of the CO₂-dependent O₂ production and delayed fluorescence of Chl a. The presence of 0.5 M NaCl in the incubation medium caused equal photoinhibition of the photosynthetic apparatus at I = 1200 μE m⁻² s⁻¹ in the mutant and wild-type cells. At I = 2400 μE m⁻² s⁻¹, we observed stronger inhibition and slower recovery of the photosynthetic apparatus in the katG ⁻ mutant than in wild-type cells. The data obtained evidence an important role of catalase-peroxidase in the system of reparation of the photosynthetic apparatus damaged by high-intensity light, especially at the background of NaCl stress.     

葡萄糖对小球藻(Chloralla sp. HN08)光合作用和生长的影响

【目的】探讨葡萄糖作为外加碳源对热带海洋小球藻(Chloralla sp.HN08)生物质生产和脂、光合色素、碳水化合物及可溶性蛋白等细胞主要成份含量的影响。【方法】分析比较小球藻HN08在光合自养和兼养(添加10 g/L葡萄糖)2种营养方式下的生长速率、细胞密度、光合放氧速率、油脂相对含量,以及可溶性总糖、淀粉和可溶性蛋白的含量。【结果】结果表明,在光照条件下葡萄糖(10 g/L)能促进小球藻(Chloralla sp.HN08)生长,提高细胞终密度,而异养条件下藻细胞逐渐衰亡。兼养条件下,细胞相对生长速率及细胞终密度分别是自养条件下的6.8倍和1.3倍。兼养藻细胞中可溶性糖、淀粉、油脂含量显著高于(P0.05)光合自养细胞,然而可溶性蛋白质和光合色素含量显著低于(P0.05)光合自养细胞。添加葡萄糖的小球藻液的光饱和点和呼吸速率均高于光自养条件下的细胞,但2种培养条件下藻液的净光合速率无显著差异(P0.05)。【结论】光照条件下,添加葡萄糖可显著提高小球藻HN08相对生长速率和细胞终密度,促进油脂与淀粉的积累。     

PHOTOACCLIMATION IN THE PHOTOTROPHIC MARINE CILIATE MESODINIUM RUBRUM (CILIOPHORA)1

Mesodinium rubrum (=Myrionecta rubra), a marine ciliate, acquires plastids, mitochondria, and nuclei from cryptophyte algae. Using a strain of M. rubrum isolated from McMurdo Sound, Antarctica, we investigated the photoacclimation potential of this trophically unique organism at a range of low irradiance levels. The compensation growth irradiance for M. rubrum was 0.5 μmol quanta · m⁻² · s⁻¹, and growth rate saturated at ∼20 μmol quanta · m⁻² · s⁻¹. The strain displayed trends in photosynthetic efficiency and pigment content characteristic of marine phototrophs. Maximum chl a–specific photosynthetic rates were an order of magnitude slower than temperate strains, while growth rates were half as large, suggesting that a thermal limit to enzyme kinetics produces a fundamental limit to cell function. M. rubrum acclimates to light‐ and temperature‐limited polar conditions and closely regulates photosynthesis in its cryptophyte organelles. By acquiring and maintaining physiologically viable, plastic plastids, M. rubrum establishes a selective advantage over purely heterotrophic ciliates but reduces competition with other phototrophs by exploiting a very low‐light niche.     

PHOSPHATE UPTAKE KINETICS IN EUGLENA GRACILIS (Z) (EUGLENOPHYCEAE) GROWN IN LIGHT/DARK CYCLES. II. PHASED PO4-LIMITED CULTURES1

The characteristics of phosphate uptake and photosynthetic capacity were studied in P-limited populations of Euglena gracilis Klebs (Z), using both P-limited batch cultures in stationary phase and cyclostat cultures grown on 14:10 LD. P uptake obeyed Michaelis-Menten kinetics between 0 and 150 μM PO₄ under both growth conditions. The value of V_max was 35% lower in the dark than in the light in the stationary phase cells. The value of K₈ was not affected by light conditions, and uptake was completely inhibited in the presence of 1 mm KCN. P uptake (at 2.0 μM PO₄) and photosynthetic capacity showed diel periodicity with peak rates occurring just before the beginning of the dark period for P uptake, and 8 h into the light period for photosynthetic capacity. V_max for P uptake increased by a factor of 1.5 over the light period, whereas K₈ remained constant at 1.4 μM PO₄. These patterns were displayed by both nondividing stationary phase cells and populations in which less than a third of the cells divided each day, indicating that the rhythmicity is not coupled to cell division.     

Adverse Effects of Epiphytic Crustose Lichens upon Stem Photosynthesis and Chlorophyll of Populus tremula L.

Dry cork layer (phellem) in stems of Populus tremula transmitted 35–55 percent of incident irradiation, depending upon moisture content. A cover of crustose Lecanora lichens reduced transmission through phellem to 10 percent or less of incident irradiation. The bark contains photosynthetically active cells. Apparent quantum yield for photosynthetic O₂-evolution was 0.017 in bark covered with dry Lecanora compared with 0.070 in naked bark. The capacity for gross photosynthesis in high light (1090 μmol photons m^?2 s^?1) was reduced by 50 percent in Lecanora-covered bark. Lecanora did not reduce the ratio between variable and maximal chlorophyll a fluorescence (F_v/F_m). Chlorophyll content per unit area was similar in leaves and naked bark of Populus tremula. The chlorophyll content in the bark decreased with increasing chlorophyll content in Lecanora. Chlorophyll a/b ratio was 2.5 in the bark compared with 4.0 in leaves and in Lecanora, and the ratio decreased down the stems. The a/b ratio was 2.3 in Lecanora covered bark compared with 2.6 in naked bark. The changes in bark photosynthesis below a Lecanora crust were probably due to acclimation of bark photosynthesis to shade, since the lichen acids in the measured lichens neither suppressed photosynthetic O₂-evolution nor changed the F_v/F_m in bark disks.     

Metabolic and photosynthetic consequences of blocking starch biosynthesis in the green alga Chlamydomonas reinhardtii sta6 mutant

Upon nutrient deprivation, microalgae partition photosynthate into starch and lipids at the expense of protein synthesis and growth. We investigated the role of starch biosynthesis with respect to photosynthetic growth and carbon partitioning in the Chlamydomonas reinhardtii starchless mutant, sta6, which lacks ADP‐glucose pyrophosphorylase. This mutant is unable to convert glucose‐1–phosphate to ADP‐glucose, the precursor of starch biosynthesis. During nutrient‐replete culturing, sta6 does not re‐direct metabolism to make more proteins or lipids, and accumulates 20% less biomass. The underlying molecular basis for the decreased biomass phenotype was identified using LC–MS metabolomics studies and flux methods. Above a threshold light intensity, photosynthetic electron transport rates (water → CO₂) decrease in sta6 due to attenuated rates of NADPH re‐oxidation, without affecting photosystems I or II (no change in isolated photosynthetic electron transport). We observed large accumulations of carbon metabolites that are precursors for the biosynthesis of lipids, amino acids and sugars/starch, indicating system‐wide consequences of slower NADPH re‐oxidation. Attenuated carbon fixation resulted in imbalances in both redox and adenylate energy. The pool sizes of both pyridine and adenylate nucleotides in sta6 increased substantially to compensate for the slower rate of turnover. Mitochondrial respiration partially relieved the reductant stress; however, prolonged high‐light exposure caused accelerated photoinhibition. Thus, starch biosynthesis in Chlamydomonas plays a critical role as a principal carbon sink influencing cellular energy balance however, disrupting starch biosynthesis does not redirect resources to other bioproducts (lipids or proteins) during nutrient‐replete culturing, resulting in cells that are susceptible to photochemical damage caused by redox stress.     

A high proportion of blue light increases the photosynthesis capacity and leaf formation rate of Rosa × hybrida but does not affect time to flower opening

Alterations in light quality affect plant morphogenesis and photosynthetic responses but the effects vary significantly between species. Roses exhibit an irradiance‐dependent flowering control but knowledge on light quality responses is scarce. In this study we analyzed, the responses in morphology, photosynthesis and flowering of Rosa × hybrida to different blue (B) light proportions provided by light‐emitting diodes (LED, high B 20%) and high pressure sodium (HPS, low B 5%) lamps. There was a strong morphological and growth effect of the light sources but no significant difference in total dry matter production and flowering. HPS‐grown plants had significantly higher leaf area and plant height, yet a higher dry weight proportion was allocated to leaves than stems under LED. LED plants showed 20% higher photosynthetic capacity (A_max) and higher levels of soluble carbohydrates. The increase in A_max correlated with an increase in leaf mass per unit leaf area, higher stomata conductance and CO₂ exchange, total chlorophyll (Chl) content per area and Chl a/b ratio. LED‐grown leaves also displayed a more sun‐type leaf anatomy with more and longer palisade cells and a higher stomata frequency. Although floral initiation occurred at a higher leaf number in LED, the time to open flowers was the same under both light conditions. Thereby the study shows that a higher portion of B light is efficient in increasing photosynthesis performance per unit leaf area, enhancing growth and morphological changes in roses but does not affect the total Dry Matter (DM) production or time to open flower.     

Growth response of cotton to CO2 enrichment in differing light environments

Experiments were conducted to examine the growth responses of cotton (Gossypium hirsutum L. cv. Coker 315) to CO₂ enrichment under different light regimes. Plants were exposed to 350 or 700 μl l^?1 CO₂ and six light treatments differing in photosynthetic period length (8 or 16 h) and in photosynthetic photon flux density (PPFD) for 32 days of vegetative growth. Higher PPFD (1 100 μmol m^?2 s^?1) was provided by a combination of high intensity discharge and incandescent lamps (HID), and lower PPFD (550 μmol m^?2 s^?1) was provided by fluorescent and incandescent lamps (F) or HID and incandescent lamps with shade cloth (HID_s). Growth was generally much slower with the 8-h photosynthetic periods, but the growth stimulation by CO₂ enrichment was larger than with 16-h photosynthetic periods. After 28 to 32 days of treatment, the growth enhancement with CO₂ enrichment was 152 and 78% for 8- and 16-h photosynthetic periods, respectively, under HID; 100 and 77% in F, and 77 and 56% in HID_s. The higher PPFD of HID positively influenced the CO₂ effect only at the slower growth rate in the 8-h light period. The stimulation of leaf area expansion by CO₂ enrichment was also greater with the 8-h photosynthetic period for all light sources. These results, and others on net assimilation rate, shoot to root dry weight ratios and specific leaf weights, suggest that the growth response to CO₂ enrichment with the longer photosynthetic period was depressed by limiting factors, perhaps nutritional, in the growth environment. The results also show that extensive variability in CO₂ response can occur under light intensities which are often used in growth chamber experiments.     

Autecology,pigment composition and toxicology of Dictyocha octonaria (Dictyochophyceae,Ochrophyta) from Wellington Harbor,New Zealand

Growth and pigment composition of both the skeleton‐bearing and naked forms and toxicology of the naked form of Dictyocha octonaria were studied here using clonal cultures derived from a single cell from Wellington Harbour. In this study both the naked and skeleton‐bearing forms showed optimum growth between 14 and 18°C. Maximum growth rates of the naked and skeleton‐bearing forms were 1.46 and 0.99 division day^?1, respectively. Growth rates of the naked form at temperatures <18°C were found to be greater than those of the skeleton‐bearing form. The reverse was true at temperatures >22°C. Cells of the naked form died at 24°C while those of the skeleton‐bearing form at 26°C. In this study the naked and skeleton‐bearing forms cultured in identical growth conditions contained almost the same pigments, except zeaxanthin, which made up about 3.2% of the total carotenoid in the skeleton‐bearing form and virtually none in the naked form. Toxicological tests conducted on rotifers indicated that the naked form was not harmful to the test organisms.     

IMPORTANCE OF MACRO‐ VERSUS MICROSTRUCTURE IN MODULATING LIGHT LEVELS INSIDE CORAL COLONIES1

Adjusting the light exposure and capture of their symbiotic photosynthetic dinoflagellates (genus Symbiodinium Freud.) is central to the success of reef‐building corals (order Scleractinia) across high spatio‐temporal variation in the light environment of coral reefs. We tested the hypothesis that optical properties of tissues in some coral species can provide light management at the tissue scale comparable to light modulation by colony architecture in other species. We compared within‐tissue scalar irradiance in two coral species from the same light habitat but with contrasting colony growth forms: branching Stylophora pistillata and massive Lobophyllia corymbosa. Scalar irradiance at the level of the symbionts (2 mm into the coral tissues) were <10% of ambient irradiance and nearly identical for the two species, despite substantially different light environments at the tissue surface. In S. pistillata, light attenuation (90% relative to ambient) was observed predominantly at the colony level as a result of branch‐to‐branch self‐shading, while in L. corymbosa, near‐complete light attenuation (97% relative to ambient) was occurring due to tissue optical properties. The latter could be explained partly by differences in photosynthetic pigment content in the symbiont cells and pigmentation in the coral host tissue. Our results demonstrate that different strategies of light modulation at colony, polyp, and cellular levels by contrasting morphologies are equally effective in achieving favorable irradiances at the level of coral photosymbionts.     

Nitrogen and carbon fixation byAnabaena sp. isolated from a rice paddy and grown under P and light limitations

Anabaena sp., isolated from a rice paddy, was investigated for its nitrogen fixation as measured by acetylene reduction activity (ARA) in P-limited continuous and light-limited semi-continuous cultures. Growth rate (μ) under P limitation was a function of cell P content (q _p). Both the photosynthetic capacity (P_max) and photosynthetic efficiency (α) increased with μ when expressed per cell, but not per unit chla. The ARA of steady-state cells under P limitation increased with μ and was linearly related to C-fixation rate. This was apparently a consequence of the control of C-fixation by P limitation. In light-limited cells, steady state ARA, both at the culture light intensity and in the dark, increased asymptotically with μ, but the activity in the dark was only about 51% of that in the light. When the light level of steady-state cells grown at a high in intensity was switched to a low level, ARA decreased exponentially with time. Dark ARA activity also showed a similar decline, but at much lower levels. Thus, ARA depended not only on light history, but also immediate photosynthesis. Steady-state ARA at the ambient intensity or in the dark showed a strong correlation with¹⁴C-fixation rate. ARA of light-limited cells showed the same light-saturation characteristics as their¹⁴C-fixation, with the same initial saturation intensity,I _k. The ratios of P_max to the maximum ARA (ARA_max), and α to the slope of ARA (α_ara) were identical. A comparison of gross to net photosynthesis and N₂ fixation suggested that there was little leakage or excretion of fixed C or N.     

Photoautotrophic Growth and Photosynthesis in Cell Suspension Cultures of Chenopodium rubrum

A method is described for growing cell suspension cultures of Chenopodium rubrum photoautotrophically for prolonged periods of time. By using a two-tier culture vessel the growth medium with the cells was separated from the CO₂ reservoir. Definite CO₂ concentrations were established by a K₂CO₃/KHCO₃ buffer. Photoautotrophic growth in C. rubrum cell suspension cultures was correlated with the CO₂ level. At 0.5% CO₂ the cell cultures contained 68 μg chlorophyll/g fresh weight and showed an increase in fresh weight of about 80% in 18 days. At 1% CO₂ an increase in fresh weight of 165% in 18 days was observed. The chlorophyll content rose up to 84 μg/g fresh weight. The photoautotrophic growth was also greatly influenced by the 2,4-D content of the medium. Cell growth was enhanced by lowering the auxin concentration. Best growth was attained (210% increase in fresh weight) at 10^?8M 2,4-D. The photosynthetic activity of the cells was measured by the light dependent ¹⁴CO₂ incorporation. At 0.5% CO₂ the cell suspensions assimilated about 100 μmol CO₂/mg chlorophyll × h. In the presence of 1% CO₂ the light driven assimilation was raised up to 185 μmol CO₂/mg chlorophyll × h. In both cases, the dark incorporation of CO₂ was merely 1.8% of the values obtained in light.