Change in Photosynthetic Capacity over the Cell Cycle in Light/Dark-Synchronized Amphidinium carteri Is Due Solely to the Photocycle

Cell cycle dependent photosynthesis in the marine dinoflagellate Amphidinium carteri was studied under constant illumination and light/dark (L/D) photocycles to distinguish intrinsic cell cycle control from environmental influences. Cells were grown in constant light and on a 14:10 L:D cycle at light intensities that would yield a population growth rate of 1 doubling per day. In the former case division was asynchronous, and cells were separated according to cell cycle stage using centrifugal elutriation. Cells grown on the L:D cycle were synchronized, with division restricted to the dark period. Cell cycle stage distributions were quantified by flow cytometry. Various cell age groups from the two populations were compared as to their photosynthetic response (photosynthetic rate versus irradiance) to determine whether or not the response was modulated primarily by cell cycle constraints or the periodic L/D cycle. Cell cycle variation in photosynthetic capacity was found to be determined solely by the L/D cycle; it was not present in cells grown in constant light.     

Photosynthetic efficiency of Chlamydomonas reinhardtii in attenuated, flashing light

As a result of mixing and light attenuation, algae in a photobioreactor (PBR) alternate between light and dark zones and, therefore, experience variations in photon flux density (PFD). These variations in PFD are called light/dark (L/D) cycles. The objective of this study was to determine how these L/D cycles affect biomass yield on light energy in microalgae cultivation. For our work, we used controlled, short light path, laboratory, turbidostat‐operated PBRs equipped with a LED light source for square‐wave L/D cycles with frequencies from 1 to 100 Hz. Biomass density was adjusted that the PFD leaving the PBR was equal to the compensation point of photosynthesis. Algae were acclimated to a sub‐saturating incident PFD of 220 µmol m^?2 s^?1 for continuous light. Using a duty cycle of 0.5, we observed that L/D cycles of 1 and 10 Hz resulted on average in a 10% lower biomass yield, but L/D cycles of 100 Hz resulted on average in a 35% higher biomass yield than the yield obtained in continuous light. Our results show that interaction of L/D cycle frequency, culture density and incident PFD play a role in overall PBR productivity. Hence, appropriate L/D cycle setting by mixing strategy appears as a possible way to reduce the effect that dark zone exposure impinges on biomass yield in microalgae cultivation. The results may find application in optimization of outdoor PBR design to maximize biomass yields. Biotechnol. Bioeng. 2012; 109: 2567–2574. © 2012 Wiley Periodicals, Inc.     

Microscopic green algae and cyanobacteria in high-frequency intermittent light

The effects of fluctuations in the irradiance onScenedesmus quadricauda, Chlorella vulgaris andSynechococcus elongatus were studied in dilute cultures using arrays of red light emitting diodes. The growth rate and the rate of photoinhibition were compared using intermittent and equivalent continuous light regimes in small-size (30 ml) bioreactors. The CO₂ dependent photosynthetic oxygen evolution rates in the intermittent and continuous light regimes were compared for different light/dark ratios and different mean irradiances. The kinetics of the electron transfer reactions were investigated using a double-modulation fluorometer. The rates of photosynthetic oxygen evolution normalized to equal mean irradiance were lower or equal in the intermittent light compared to the maximum rate found in the equivalent optimal continuous light regime. In contrast, the growth rates in the intermittent light can be higher than the growth rate in the equivalent continuous light. Photoinhibition is presented as an example of a physiological process affecting the growth rate that occurs at different rates in the intermittent and equivalent continuous lights. The difference in the dynamics of the redox state of the plastoquinone pool is proposed to be responsible for the low photoinhibition rates observed in the intermittent light.     

Photoperiod influences endogenous indoleamines in cultured green alga Dunaliella bardawil

Effect of light intensity and photoperiod on growth, indoleamines and carotenoid production was studied in unicellular green algae D. bardawil. Maximum biomass and carotenoid contents were found when cultures were grown in light (intensity of 2.0 Klux) at a photoperiod of 16/8h light and dark cycle. There was a profound influence of tested photoperiod conditions of light:dark viz. 8:16, 10:14, and 12:12 hr, continuous light on indoleamines (SER and MEL) production as estimated by HPLC and confirmed by mass spectral data obtained from LC-MS-ESI studies. Serotonin level increased from 908 to 1765 pg/g fresh wt with increase in light duration and melatonin level increased from 267 to 584 pg/g fresh wt during increase in dark phase. Carotenoids production was high in continuous light than other tested conditions.     

光环境对胡桃楸幼苗生长与光合作用的影响

为了解胡桃楸幼苗对光的需求及适应规律,采用Li-6400便携式光合测定系统研究了不同光环境处理（100%、60%、30％和15%自然光）条件下3年生胡桃楸幼苗（适应1年后）叶片光合能力的季节变化及其对光强的响应.结果表明：在春季,胡桃楸幼苗对光反应不敏感,夏季和秋季随着光强的增加,叶片的最大光合速率、最大羧化速率和最大电子传递速率均显著增加（P<0.05）.光饱和点随光强的下降而降低（P<0.05）,表观量子效率、暗呼吸速率和光补偿点在不同光环境下未发现显著差异.100%和60%自然光处理的幼苗相对生长率差异不显著,但是随着光强下降,相对生长率显著下降（P<0.05）,为60%>30％>15%自然光处理.胡桃楸幼苗对不同的光环境表现出较强的适应性和可塑性;同时,通过降低光饱和点和减少碳积累,也能适应15%～30％自然光环境.     

EXTREMELY HIGH GROWTH RATE OF THE SMALL DIATOM CHAETOCEROS SALSUGINEUM ISOLATED FROM AN ESTUARY IN THE EASTERN SETO INLAND SEA,JAPAN1

Small single‐celled Chaetoceros sp. are often widely distributed, but frequently overlooked. An estuarine diatom with an extremely high growth potential under optimal conditions was isolated from the Shinkawa‐Kasugagawa estuary in the eastern part of the Seto Inland Sea, western Japan. It was identified as Chaetoceros salsugineum based on morphological observations. This strain had a specific growth rate of 0.54 h^?1 at 30°C under 700 μmol · m^?2 · s^?1 (about 30% of natural maximal summer light) with a 14:10 L:D cycle; there was little growth in the dark. However, under continuous light it grew at only 0.35 h^?1 or a daily specific growth rate of 8.4 d^?1. In addition, cell density, chlorophyll a, and particulate organic carbon concentrations increased by about 1000 times in 24 h at 30°C under 700 μmol · m^?2 · s^?1 with a 14:10 L:D cycle, showing a growth rate of close to 7 d^?1. This very rapid growth rate may be the result of adaptation to this estuarine environment with high light and temperature. Thus, C. salsugineum can be an important primary producer in this estuary in summer and also an important organism for further physiological and genetic research.     

INFLUENCE OF LOW LIGHT AND A LIGHT: DARK CYCLE ON NO3– UPTAKE,INTRACELLULAR NO3–, AND NITROGEN ISOTOPE FRACTIONATION BY MARINE PHYTOPLANKTON1

The nitrogen isotope enrichment factor (ɛ) of four species of marine phytoplankton grown in batch cultures was determined during growth in continuous saturating light, continuous low light, and a 12:12‐h light:dark cycle, with nitrate as a nitrogen source. The low growth rate that resulted from low irradiance caused an increased accumulation of the intracellular nitrate pool and/or a reduction in cell volume and was correlated to a species‐specific increase in the measured ɛ value, compared with the saturating light conditions. The largest response was in the diatom Thalassiosira weissflogii (Grun.) Fryxell et Hasle, which showed a nearly 3‐fold increase between high and low light conditions (6.2–15.2‰). The smallest response was in T. pseudonana (Hustedt) Hasle et Heimdal, which showed no change in the ɛ value of approximately 5‰ in both high and low light conditions. There was significant but smaller increases in the ɛ value for the diatom T. rotula Meunier (2.7–5.6‰) and the prymnesiophyte Emiliania huxleyi (Lohm.) Hay et Mohler (4.5–9.4‰) between high and low light levels. In the light:dark experiments, all three diatoms but not the prymnesiophyte exhibited an increase in ɛ. This increase was linked to the ability of diatoms to assimilate nitrate at night. The results of the these experiments suggest that the light regime influences the relative uptake, assimilation, and efflux rates of nitrate and results in differences in the expression of the isotope effect by the enzyme nitrate reductase. Therefore, variations in nitrate isotope fractionation in nature can be more accurately interpreted when the light regime and species composition are taken into consideration.     

Pea seedling growth and development regulated by cotyledons and modified by irradiance

Growth and development of hydroponically grown pea seedlings ( Pisum sativum L. cv. Alaska) were measured using stem and root length as well as number of leaves and lateral roots. The growth was dependent on the presence of cotyledons and was modulated by the irradiance. All plants were grown in a full nutrient solution. If grown at low irradiance (73 μmol m^-2s^-1) they depended more and for a longer time on the cotyledons than plants grown at high irradiance (220 μmol m^-2s^-1). Low irradiance caused stem elongation but decreased root length and number of lateral roots as compared to plants grown at high irradiance. The dark respiration of the leaves was measured as oxygen uptake. In plants grown at the low irradiance, excision of the cotyledons caused the rate of oxygen uptake to increase by a factor of three, and the increase was sensitive to cyanide. Decotyledonized plants showed a high respiration rate and a diminished leaf growth for their entire life cycle. CO₂ fixation also increased in decotyledonized pea seedlings grown at either irradiance. The mobilization of food reserves from the seeds was positively correlated to seed dry weight, but only if the plants were grown at 73 μmol m^-2s^-1. Increasing dry weight of the seed enhanced top growth, whereas root growth was depressed, so that top and root responds differently with regard to that part of growth which depends on mobilization of reserves from the seed.     

The Calvin cycle in cyanobacteria is regulated by CP12 via the NAD(H)/NADP(H) ratio under light/dark conditions

In Synechococcus PCC7942 cells grown in the dark, the concentrations of NAD(H) and NADP(H) were 128+/-2.5 and 483+/-4.0 microm, respectively, while those in the cells under light conditions were 100+/-5.0 and 649+/-7.0 microm, respectively. Analysis of gel filtration indicated that the change of the ratio of NADP(H) to NAD(H) in cyanobacterial cells under light/dark conditions controls the reversible dissociation of the PRK/CP12/GAPDH complex (approximately 520 kDa) consisting of phosphoribulokinase (PRK), CP12, and glyceraldehyde-3-phosphate dehydrogenase (GAPDH). S. 7942 CP12 lacked the two Cys residues essential for formation of the N-terminal peptide loop in the CP12 of higher plants, but the N-terminal region of S. 7942 CP12 had the ability to be associated with PRK. The growth of mutant cells in which the CP12 gene was disrupted by a kanamycin resistance cartridge gene was almost the same as that of wild-type cells under continuous light conditions. However, under the light/dark cycle (12 h/12 h), the growth of CP12-disrupted mutant cells was significantly inhibited compared with that of wild-type cells. The mutant cells showed a decreased rate of O2 consumption and an increased level of ribulose 1,5-bisphosphate compared with wild-type cells in the dark. These data suggest that under light and dark conditions, the oligomerization of CP12 with PRK and GAPDH regulates the activities of both enzymes and thus the carbon flow from the Calvin cycle to the oxidative pentose phosphate cycle.     

Efficient utilisation of high photon irradiance for mass production of photoautotrophic micro-organisms

Basic issues involved in effective use of a high photon irradiance for mass production of microalgae are elucidated: efficient utilisation of high irradiance requires cultures of high cell density grown in reactors with a narrow light path. The smaller the light-path, the higher the growth rate and the volume output rate (g L⁻¹d⁻¹) of cell mass. Areal productivity (g m⁻²d⁻¹) may be inversely related to the length of light-path (e.g. Spirulina platensis) or directly related to it, as is the case with Nannochloropsis sp., in which the areal output rate increased with the increase in the light-path and the areal volume (L m⁻²). Inhibition of cell growth in Nannochloropsis became evident as cell concentration increased above a certain point. Response in cell growth to elevated irradiance was therefore possible only when the growth medium of ultrahigh cell density cultures was frequently changed. Inhibitory activity to culture growth may be directly involved in determining the optimal cell density (which results in the highest output of cell mass) and hence the optimal light-path. Under optimal growth conditions, cultures of high cell densities responded well to the rate of stirring, the relative beneficial effect of mixing increasing with the increase in cell density. This revised version was published online in June 2006 with corrections to the Cover Date.     

The role of polyglucose in oxygen-dependent respiration by a new strain of Desulfovibrio salexigens

Abstract: Desulfovibrio salexigens strain Mastl was isolated from the oxic/anoxic interface of a marine sediment. Growth under sulfate-reducing conditions was accompanied by polyglucose accumulation in the cell with every substrate tested. Highest polyglucose storage was found with glucose (0.8–1.0 g polyglucose (g protein)⁻¹), but the growth rate with this substrate was very low (0.015 h⁻¹). Anaerobically grown cells of strain Mastl exhibited immediate oxygen-dependent respiration. The endogenous oxygen reduction rate was proportional to the polyglucose content. The rate of aerobic respiration of pyruvate was also directly related to the polyglucose content indicating that this organism was only able to respire with oxygen as long as polyglucose was present. Maximum oxygen reduction rates were found at air saturating concentrations and were relatively low (3–50 nmol O₂ min⁻¹ (mg protein)⁻¹). Catalase was constitutively present in anaerobically grown cells. When batch cultures were exposed to oxygen, growth ceased immediately and polyglucose was oxidized to acetate within 40–50 h. Like the oxygen reduction activity, the nitro blue tetrazolium (NBT)-reduction activity in these cells was proportional to the polyglucose content. Under anaerobic starvation conditions there was no correlation between the NBT-reduction activity and polyglucose concentration and polyglucose was degraded slowly within 240 h. The ecological significance of aerobic polyglucose consumption is discussed.     

Effect of light/dark cycles on wastewater treatments by microalgae

Chlorella kessleri was cultivated in artificial wastewater using diurnal illumination of 12 h light/12 h dark (L/D) cycles. The inoculum density was 10⁵ cells/mL and the irradiance in light cycle was 45 μmol m² s⁻¹ at the culture surface. As a control culture, another set of flasks was cultivated under continuous illumination. Regardless of the illumination scheme, the total organic carbon (TOC) and chemical oxygen demand (COD) was reduced below 20% of the initial concentration within a day. However, cell concentration under the L/D lighting scheme was lower than that under the continuous illuminating scheme. Thus the specific removal rate of organic carbon under L/D cycles was higher than that under continuous illumination. This result suggested thatC. kessleri grew chemoorganotrophically in the dark periods. After 3 days, nitrate was reduced to 136.5 and 154.1 mg NO₃ ⁻-N/L from 168.1 mg NO₃ ⁻-N/L under continuous illumination and under diurnal cycles, respectively. These results indicate nitrate removal efficiency under continuous light was better than that under diurnal cycles. High-density algal cultures using optimized photobioreactors with diurnal cycles will save energy and improve organic carbon sources removal.     

INTERACTIONS OF LIGHT/DARK CYCLES AND NITROGEN PULSES ON THE TIMING OF CELL DIVISION IN THE NITROGEN-LIMITED MARINE DIATOM CYLINDROTHECA FUSIFORMIS (BACILLARIOPHYCEAE)1

Cell division in most eukaryotic algae grown on alternating periods of light and dark (LD) is synchronized or phased so that cell division occurs only during a restricted portion of the LD cycle. However, the phase angle of the cell division gate, the time of division relative to the beginning of the light period, is known to be affected by growth conditions such as nutrient status and temperature. In this study, it is shown that the phase angle of cell division in a diatom, Cylindrotheca fusiformis Reimann and Lewin, is affected by the N-limited growth rate; cell division occurred later in the dark period (12:12 h LD cycle) when the growth rate was infradian (D = 0.42 d^?1) than when it was ultradian (D = 1.0 d^?1). Nitrogen-pulses did not affect the phase angle of the division gate, but could shift the time of peak cell division activity within the division gate. The effects, if any, of N-pulses were dependent upon the growth rate and the time of day that the pulses were administered. These responses indicate that the timing of cell division in this diatom is not determined solely by the zeitgeber from the LD cycle, but rather that a LD cycle control mechanism and a N-mediated control mechanism are both involved and are somewhat interdependent. In addition, an increase in protein was observed immediately after administering a N-pulse to C. fusiformis in the ultradian growth mode indicating that the accumulation of protein can be uncoupled from the cell division cycle.     

A comparative study of the responsivity of Sinapis alba L. seedlings to pulsed and continuous irradiation

Anthocyanin formation in 36h dark grown Sinapis alba L. seedlings and inhibition of hypocotyl elongation in 36h and 54h dark grown and 54h and 7 day light grown seedlings in response to continuous red light could be substituted for by hourly 5 min light pulses where the total fluence over the irradiation period is the same. These pulses are partially (36h) or almost totally (54h and 7 day) reversible by subsequent far-red (RG 9) light pulses. In contrast to 654 nm light, hourly light pulses with 552 nm, 449 nm and 715 nm can at best only partially substitute for continuous irradiation. These data are discussed with respect to the commonly used models for the phytochrome high irradiance response.Abbreviations P_tr tar-red absorbing form of phytochrome - SAN 9789 4-chloro-5-(methyl-amino)-2-(,,-trifluoro-m-tolyl)-3(2H)-pyridazinone=Norflurazon - HIR High irradiance response     

The flexible interrelation between AOX respiratory pathway and photosynthesis in rice leaves.

Alternative respiratory pathway was investigated in rice seedlings grown under total darkness, light/dark cycle, or continuous light. The capacity of the alternative pathway was relatively higher in leaves that had longer light exposure. An analysis of rice AOX1 multigene family revealed that AOX1c, but not AOX1a and AOX1b, had a light-independent expression. The alternative oxidase (AOX) inhibitor, salicylhydroxamic acid (SHAM, 1mM), inhibited nearly 68% of the capacity of the alternative pathway in leaves grown under different light conditions. The plants grown under different light periods were treated with SHAM and then were exposed to illumination for 4h. The transition from dark to 4h of light stimulated the capacity of alternative pathway in etiolated rice seedlings and in those grown under light/dark cycle, whereas the capacity of the alternative pathway was constant in seedlings grown under continuous light with additional 4h of illumination. Etiolated leaves did not show any CO(2) fixation after 4h of illumination, and the increase in chlorophyll content was delayed by the SHAM pretreatment. When seedlings grown under light/dark cycle were moved from dark and exposed to 4h of light, increases in chlorophyll content and CO(2) fixation rate were reduced by SHAM. Although these parameters were stable in plants grown under continuous light, SHAM decreased CO(2) fixation rate but not the chlorophyll content. These results indicate that the role and regulation of AOX in light are determined by the developmental stage of plant photosynthetic apparatus.     

Effect of photoperiod on growth and feed conversion of juvenile wild carp, Cyprinus carpio

Because of the effect of photoperiod on physiological and biochemical processes in fish, this study aimed to evaluate the effect of manipulated photoperiod on growth, feed conversion and survival of wild carp, Cyprinus carpio. Fish received six photoperiod regimes (light:dark cycle) including: natural photoperiod (control), 24L:0D, 16L:8D, 12L:12D, 8L:16D and 0L:24D by the three replications. Regulated photoperiods as a 16L:8D or 12L:12D light/dark cycle significantly improved growth rate and food conversion ratio of wild carp.     

Influence of Melatonin on the Rate of Rana pipiens Tadpole Metamorphosis In Vivo and Regression of Thyroxine-Treated Tail Tips In Vitro

Metamorphosis of Rana pipiens tadpoles may be retarded when the light phase of the light/dark (LD) cycle is shortened or when thyroxine (T₄) is given in the dark because melatonin peaks during the dark. Injection of premetamorphic tadpoles in spontaneous metamorphosis with melatonin (15 μg) retarded tail growth and hindlimb development on 18L:6D but had no significant effect on 6L:18D. During induced metamorphosis (30 μg/liter T₄), melatonin injections retarded tail resorption on 18L:6D and accelerated it on 6L:18D, but did not affect the hindlimb. When melatonin was injected during T₄ immersion at different times in the photophase on 18L:6D (L onset 0800 hr), tail regression was retarded by melatonin at 1430 or 2030 hr. At 0830 hr, shrinkage of tail length was accelerated whereas tail height was not affected. Tail tips in vitro induced to resorb by 0.2 μg/ml T₄ in Niu-Twitty solution regressed more slowly in the presence of melatonin (10 or 15 μg/ml) than with T₄ alone on both 6L:18D and 18L:6D. The findings implicate melatonin in LD cycle effects on tadpole metamorphic rate in vivo , show the importance of the time of melatonin injections, and indicate that melatonin antagonizes the metamorphic action of T₄ at the tissue level.     

DIEL VARIATIONS OF CARBOHYDRATES AND NEUTRAL LIPIDS IN NITROGEN‐SUFFICIENT AND NITROGEN‐STARVED CYCLOSTAT CULTURES OF ISOCHRYSIS SP.1

The goal of this study was to investigate the time response of two major carbon (C) reserves, respectively neutral lipids (NL) and total carbohydrate (TC), in the Haptophyte Isochrysis sp. growing in nitrogen (N)‐sufficient or N‐starved conditions and under light:dark (L:D) cycles. Experiments were carried out in a cyclostat culture system that allowed the following of the dynamics of the main cell compounds at both hourly and daily time scales. Under N‐sufficient conditions, the L:D cycles cause the population to be synchronized, with most of the cells dividing at the beginning of the dark period. The C‐specific growth rate was maximal around midday and negative during the dark period due to respiration processes. NL and TC both accumulated during the day and consumed during the night. We showed that NL and TC are highly dynamic compounds, as more than three quarters of NL and TC accumulated during the light period were consumed during the dark period. In contrast to NL, phospholipid and glycolipid to C ratios remained quite stable during the light/dark cycles. The major effect of N starvation on the NL and TC dynamics was to uncouple their diel variations from the L:D cycle, in two different ways depending on their respective role during short‐term acclimation. Whereas the TC per cell ratio increased rapidly to reach a stable value in response to N starvation, NL per cell continued to oscillate, but with a pattern out of phase with the L:D cycle.     

Outdoor production of Phaeodactylum tricornutum biomass in a helical reactor

The production of microalga Phaeodactylum tricornutum in an outdoor helical reactor was analysed. The influence of temperature, solar irradiance and air flow rate on the yield of the culture was evaluated. Biomass productivities up to 1.5 g l(-1) per day and photosynthetic efficiency up to 14% were obtained by maintaining the cultures below 30 degrees C, dissolved oxygen levels less than 400% Sat. (with respect to air saturated culture) and controlling the cell density in order to achieve an average irradiance within the culture below 250 microE m(-2) s(-1). Under these conditions, the fluorescence parameter, Fv/Fm, which reflects the maximal efficiency of PSII photochemistry, remained roughly 0.6-0.7 and growth rates up to 0.050 h(-1) were achieved. The average irradiance and the light/dark cycle frequency, were the variables determining the behaviour of the cultures. A hyperbolic relationship between growth rate and biomass productivity with the average irradiance was observed, whereas both biomass productivity and photosynthetic efficiency linearly increased with the light/dark cycle frequencies. Optimum design and operational conditions which maximise the production of P. tricornutum biomass in outdoor helical reactors were determined.     

5种光周期对锦鲤生长、能量收支及生物钟基因表达的影响

研究不同光照周期(长光照18L﹕6D、短光照8L﹕16D、连续光照24L﹕0D、连续黑暗0L﹕24D和对照组12L﹕12D)条件对锦鲤(Cyprinus carpio)生长、能量收支及生物钟基因相对表达量的影响, 实验周期为90d。结果表明, 在整个试验周期, 各光周期处理组成活率无显著变化(P＞0.05), 24L组和18L﹕6D组饲料系数显著低于12L﹕12D组、8L﹕16D组和24D组(P＜0.05)。终末体重、特定生长率在实验前30d内受光照影响不大(P＞0.05), 而实验30d后出现显著变化, 延长光周期尤其是连续光照会显著增加终末体重和特定生长率(P＜0.05); 对比连续黑暗0L﹕24D组, 其他光周期处理组锦鲤摄食能分配在代谢和生长的比例显著增加(P＜0.05); 4个生物钟相关Clock、Per2、Cry1和Bmal1基因相对表达水平无明显规律。综上所述, 延长光照时间, 尤其是连续光照有利于锦鲤幼鱼的生长和发育。