Influence of blue light on the activity of phosphofructokinase in Chlorella kessleri

In crude extracts of Chlorella kessleri Fott and Novákóva cells grown autotrophically in white light the activity of phosphofructokinase (PFK, EC 2.7.1.11) is 62.9 ± 1.5 nmol (mg protein)⁻¹ min⁻¹ under optimized test conditions. It is greatly increased in red [88.3 ± 1.8 nmol (mg protein)⁻¹ min⁻¹], but somewhat decreased [57.0 ± 0.5 nmol (mg protein)⁻¹ min⁻¹] in blue light of equal productivity. Mixtures of blue and red light yield the low activity as long as blue light represents at least 35% of the total quantum fluence rate. The rough wavelength dependence of the counteracting effect of short wavelength light on the increasing effect of red light exhibits a broad peak at 460 nm, reminiscent of action spectra of the blue/UV photoreceptors(s). Upon transfer of red light-grown cells to blue light, the decrease develops slowly within 72 h; it cannot be prevented by 3-(3,4-dichlorophenyl)-1,1-dimethyl urea (DCMU). Since there is less carbohydrate in blue than in red light-exposed cells, correlations between biosynthesis of PFK and level of carbohydrate are discussed, based on the assumption that red light decreases and/or blue light increases the transport of metabolites across the chloroplast envelope.     

Properties of Oligomeric Forms of Phosphofructokinase from Chlorella kessleri Grown under Different Light Conditions

Fast protein liquid chromatography on Superose 6 of crude extracts from Chlorella kessleri, Fott et Novákóva, grown autotrophically in blue or in red light yields three different oligomeric forms of phosphofructokinase (PFK, EC 2.7.1.11). Their substrate affinities and responses to homotropic and heterotropic effectors are different. In vitro, the degree of oligomerization of the enzyme can be influenced by specific intermediates or cofactors. Its substrate, MgATP (10 mM/5 mM), and the negative effector, phosphoenolpyruvate (5 mM), both lead to some dissociation, while the second substrate, fructose-6-phosphate (5 mM), and the positive effector, inorganic phosphate (50 mM), have no effect. It is discussed whether formation or dissociation of oligomeric PFK forms in vivo result from alterations in the levels or in the intracellular distribution of effector molecules and whether such processes are involved in the different regulation of cell metabolism in blue or in red light.     

Influences of Blue and Red Light on the Photosynthetic Apparatus of Chlorella kessleri*

In white light of 33.2 μmol . m^?2 . s^?1 oxygen evolution of Chlorella kessleri is about 30 % higher after growth in blue light than after growth in red light of the same quantum fluence rate. When determined by the light-induced absorbance change at γ 820 nm, blue light-adapted cells possess about 60% more reaction centres per total chlorophyll in photosystem II. Correspondingly, the cells exhibit about 30% more Hill activity of PS II. Conversely, red light-adapted cells contain relatively more reaction centres and higher electron flow capacities of photosystem I. The distribution of total chlorophyll among the pigment-protein complexes, CPI, CPIa, CPa, and LHC II, corresponds to these data. There is more chlorophyll associated with the light-harvesting complex of PS II, LHC II, in cells under blue light conditions, but more chlorophyll bound to both complexes of PS I, CPI and CPIa, in cells under red light conditions. The respective ratios of chlorophyll a/chlorophyll b of all complexes are identical for blue and red light-adapted cells. This results in a higher relative amount of chlorophyll b in blue light-adapted cells. Total carotenoids per total chlorophyll are increased by 20% in red light-adapted cells. Their distribution among the pigment-protein complexes is unknown, however the ratios of lutein, neoxanthin and violaxanthin extractable from LHC II are different in blue (32.1:35.9:32.0) and in red (51.4:26.7:21.9) light-adaptod cells.     

Blue and Red Light-dependent Alterations in the Ratio of two Forms of Glutamine Synthetase in Chlorella kessleri

Two forms of glutamine synthetase (EC 6.3.1.2) can be separated in crude extracts of Chlorella kessleri on the basis of their different surface charges. The two enzyme forms (GS1 and GS2) respond differently upon transferring the cells from darkness to autotrophic growth in white light: the activity of GS2 increases, that of GS1 remains unchanged. The increase in GS2 activity is only brought about by blue light; in red light GS2 activity appears to be uninfluenced, while that of GS1 increases. There are no indications of wavelength-dependent oligomerization processes as a cause for the observed activity alterations. There is however, a strong influence of inhibitors of protein biosynthesis. Cycloheximide and lincomycin both affect the blue light-dependent increase in activity of GS2, cycloheximide preventing that of GS1 in red lgiht completely. Since literature data point to localization of GS2 in the chloroplast, and GS1 in the cytosol, the data are discussed in view of two different photoreceptors involved in the regulation of the amounts of GS1 and GS2 in different compartments of the Chlorella cell.     

Oligomeric Forms of Glycolytic Enzymes in Chlorella Grown in Different Light Qualities*

Fast protein liquid chromatography on Superose 6 of crude extracts from the green alga Chlorella kessleri cultivated autotrophically in white light reveals several peaks with phosphofructokinase (PFK, EC 2.7.1.11) or pyruvate kinase (PK, EC 2.7.1.40) activity with molecular weights larger than the usually reported ones of 320–380 and 240 kDa, respectively. All other glycolytic enzymes are eluted as one peak each with a molecular weight corresponding to data from the literature. Indirect evidence indicates that the various forms of PFK and presumably PK are oligomers. The occurrence of different PFK species depends markedly on growth conditions such as wavelength of light: Red light leads to only one rather large PFK (1,580 kDa), blue light to two smaller species (760 and 360 kDa). All species are probably present in white light-grown cells (1,500, 1,050, 930, 700 and 440 kDa). The various light qualities do not significantly affect all other glycolytic enzymes. PK constantly exhibits four forms with molecular weights of 830, 680, 480, 305 kDa. Experiments with the chlorophyll-free mutant no. 20 of Chlorella kessleri support the assumption that oligomerization of enzymes is characteristic of regulatory enzymes, thereby providing the cell with an additional regulatory means.     

Expression of eukaryotic plasma membrane transporter HUP1 from Chlorella kessleri in Escherichia coli

To study the effect of sterols on the activity of the eukaryotic plasma membrane transporter, the hexose-proton symporter HUP1 from the unicellular alga Chlorella kessleri was expressed in Escherichia coli, a prokaryotic microorganism containing virtually no sterols. Under certain conditions, the recombinant protein was partially active in this prokaryotic organism. The heterologously produced HUP1p was purified from membrane fractions of E. coli and reconstituted in an in vitro system. The presence of ergosterol during solubilization, purification and reconstitution resulted in an increased activity of the reconstituted protein. Its activity, however, was 5-6 times lower as compared to the activity of HUP1p produced in Saccharomyces cerevisiae membranes and solubilized, purified, and reconstituted under the same conditions as above.     

The effect of SAN 9789 and light on the germination of seeds from Taraxacum vulgare

Photoblastic seeds (achenes) of Taraxacum vulgare coll. were treated with a water solution of SAN 9789, 4-chloro-5 (methylamino) -2- (α,α,α-trifluoro- m -tolyl) -3(2H) pyridazinone. SAN-treatment increased the germination in darkness from 0 to 12%. An irradiation for 5 min with red light, giving a germination of 12% for seeds in water only, gave together with SAN treatment a germination of 60%. In both water and SAN, the effect of red irradiation could be reversed by a short irradiation (15 min) of far-red light. If far-red light was repeatedly given (5 min per h) it had hardly any effect on germination in water (4% germination), but for seeds in SAN solution, intermittent far-red light had a stimulating effect (63% germination). If far-red light was given continuously for 96 h, the germination in water was 1% and in SAN solution 17%. The results in the present paper indicate that SAN may broaden the concentration interval of P_fr for which germination is high.     

Effects of light flashes on the dark closure of Albizzia julibrissin pinnules

An electronic flash unit is used to deliver, at the beginning of a 10 min dark period and within a few ms, large doses of light to Albizzia julibrissin pinnules, to ascertain their effects on the rate of pinnule closing. In a series of alternating light flashes at 710 and 550 nm, the first 710 nm light flash significantly retards closing. A following light flash at 550 nm negates the far-red induced delay. The second 710 nm light flash delays closing less effectively than the first when given within 4 s after the green flash, but is just as effective when given after 30 s. The delay brought about by the second 710 nm light flash is again abolished by a light flash at 550 nm. A light flash at 660 nm has no effect on pinnule closing by itself and is also ineffective in reversing the far-red induced delay. A series of ten 710 nm light flashes becomes most effective in delaying closure when there is a dark interval of one min between flashes. The closing delay induced by a 710 nm light flash escapes reversal by a 550 nm light flash when the dark interval between the two flashes exceeds 2–3 min. A 750 nm light flash has no retarding effect on pinnule closing, but it becomes effective when preceded by a 660 nm or 550 nm light flash. The results obtained are suggested to be due to light absorbed by phytochrome and an unknown photoreceptor with green, far-red photoreversal property.     

Phytochrome A is an irradiance-dependent red light sensor

Plants perceive red (R) and far-red (FR) light signals using the phytochrome family of photoreceptors. In Arabidopsis thaliana, five phytochromes (phyA-phyE) have been identified and characterized. Unlike other family members, phyA is subject to rapid light-induced proteolytic degradation and so accumulates to relatively high levels in dark-grown seedlings. The insensitivity of phyA mutant seedlings to prolonged FR and wild-type appearance in R has led to suggestions that phyA functions predominantly as an FR sensor during the early stages of seedling establishment. The majority of published photomorphogenesis experiments have, however, used <50 micromol m(-2) sec(-1) of R when characterizing phytochrome functions. Here we reveal considerable phyA activity in R at higher (>160 micromol m(-2) sec(-1)) photon irradiances. Under these conditions, plant architecture was observed to be largely regulated by the redundant actions of phytochromes A, B and D. Moreover, quadruple phyBphyCphyDphyE mutants containing only functional phyA displayed R-mediated de-etiolation and survived to flowering. The enhanced activity of phyA in continuous R (Rc) of high photon irradiance correlates with retarded degradation of the endogenous protein in wild-type plants and prolonged epifluorescence of nuclear-localized phyA:YFP in transgenic lines. Such observations suggest irradiance-dependent 'photoprotection' of nuclear phyA in R, providing a possible explanation for the increased activity observed. The discovery that phyA can function as an effective irradiance sensor, even in light environments that establish a high Pfr concentration, raises the possibility that phyA may contribute significantly to the regulation of growth and development in daylight-grown plants.     

Trehalose phosphate synthase overexpression in Parachlorella kessleri improves growth and photosynthetic performance under high light conditions

Parachlorella kessleri is a promising oil-bearing marine alga which shows decreased growth under high light stress. Osmolytes are known to relieve stress by protecting the cell membrane, proteins, and enzymes. Enhanced production of osmolyte (trehalose) was thus used to relieve stress in P. kessleri by overexpression of trehalose phosphate synthase (TPS) gene. Transformed P. kessleri was grown under different light regimes to study the effect of trehalose overproduction on growth. Study of one of the TPS transformants showed increased trehalose as well as increased biomass and decreased pigments, reactive oxygen species, and lipid peroxidation of cell membrane. The improved photosynthetic performance of the transformant was also signified by pulse-amplitude-modulated fluorometric analysis. All of these factors reveal improved stress tolerance under high light conditions by increased trehalose accumulation due to TPS overexpression in P. kessleri.     

Effects of red light on the fluence–response relationship for pulse-induced phototropism of maize coleoptiles

Dark-adapted coleoptiles of maize (Zea mays L.) were treated with red light (3min at 10.5 μmol m^?2S^?1) and were Stimulated, after a dark interval, with a pulse of unilateral blue light to induce phototropism. Phototropic fluence-response curves were obtained in this way for different dark intervals. It was confirmed that the bell-shaped fluence-response curve for the first pulse-induced positive phototropism (FPIPP) shifts to higher fluences following the red-light treatment, the maximal shift being achieved at a dark interval of 2h. We found, however, that the two arms of the Fluence-response curve do not shift synchronously. The shift of the descending arm to higher fluences began at 15 min. The ascending arm showed a slight shift to lower fluences before a greater shift to higher flucnces. the change of the shift direction occurring at 30–40min. Accordingly, the fluence-response curve obtained for a 30 min dark interval was comparatively wide. Although dark-adapted coleoptiles showed only fPIPP, another bell-shaped fluence-response curve, representing the second pulse-induced positive phototropism (sPIPP), appeared gradually after the red-light treatment. These changes of the phototropic fluence– respnse curve following exposure to red light are likely to have adaptive values because they favour phototropism under brighter light.     

蛋白核小球藻光驯化的快速光曲线变化

通过测量快速光曲线研究了强光和弱光驯化对蛋白核小球藻(Chlorella pyrenoidosa)光合作用的影响。弱光驯化后的初始斜率α高于强光驯化后,而半饱和光强I_k明显低于强光驯化后,表明弱光驯化提高了蛋白核小球藻的捕光能力。强光驯化后最大光合速率P_m高于弱光驯化后,而光抑制参数β小于弱光驯化后,表明强光驯化提高了蛋白核小球藻的光合能力和对强光的耐受性。     

Investigation and modeling of the effects of light spectrum and incident angle on the growth of Chlorella vulgaris in photobioreactors

An in‐depth investigation of how various illumination conditions influence microalgal growth in photobioreactors (PBR) has been presented. Effects of both the light emission spectrum (white and red) and the light incident angle (0° and 60°) on the PBR surface were investigated. The experiments were conducted in two fully controlled lab‐scale PBRs, a torus PBR and a thin flat‐panel PBR for high cell density culture. The results obtained in the torus PBR were used to build the kinetic growth model of Chlorella vulgaris taken as a model species. The PBR model was then applied to the thin flat‐panel PBR, which was run with various illumination conditions. Its detailed representation of local rate of photon absorption under various conditions (spectral calculation of light attenuation, incident angle influence) enabled the model to take into account all the tested conditions with no further adjustment. This allowed a detailed investigation of the coupling between radiation field and photosynthetic growth. Effects of all the radiation conditions together with pigment acclimation, which was found to be relevant, were investigated in depth. © 2016 American Institute of Chemical Engineers Biotechnol. Prog., 32:247–261, 2016     

不同光质对小球藻光自养培养积累油脂的影响

研究了5种光质对小球藻(Chlorella vulgaris)M209256生长和产油的影响。结果表明:蓝光为小球藻的最适生长和产油光质;与其他光质相比,蓝光培养的小球藻生物量和油脂含量均较高,为2.40×107个/mL和28%;红光培养的小球藻生长最慢且油脂含量最低,为1.32×107个/mL和15.13%,表现出明显的"红降"现象。在GCMS分析的基础上,对油脂甲酯化后的十六烷值进行评估,结果发现:蓝光的十六烷值最高;5种光质培养的小球藻所产油脂,甲酯十六烷值均在47以上。因此,小球藻油脂所制备的生物柴油具有较好的燃烧性能。     

棕背黑头鸫繁殖习性初步观察

2008年和2009年的4～8月,在四川省雅江县帕姆岭对棕背黑头鸫Turdus kessleri的繁殖生态进行了初步观察.该鸟繁殖期在4月下旬至7月上旬,营树上巢,营巢树种为高山栎Quercus aquifolioides和鳞皮冷杉Abies squamata.窝卵数为2~3枚(n=7),平均卵重(7.96±0.03)g(n=8),卵长径(32.7±0.17)mm,短径(21.9±0.13)mm(n=13),雌雄共同孵卵,以雌性为主,孵化期为15~17 d(n=2),孵化率为83.3%(n=18).雌雄共同育雏,以雄性为主,雏鸟出飞后主要在巢周围的林下或灌从活动,这时亲鸟仍会对幼鸟喂食.在同一繁殖季对巢有重复利用的现象.     

The effects of SAN 9789 and light on phytochrome and the germination of lettuce seeds

Photoblastic seeds (akenes) of lettuce (Lactuca sativa (L.) cv. Grand Rapids) were treated with SAN 9789 [4-chloro-5-(methylamine)-2-a, a, a,-trifluoro-m-tolyl-3-(2H)-pyridasinone]. The seeds weere placed in Petri dishes on filter paper soaked with water or SAN solution. The treatment increased the germination in darkness from 17% for water to 78% for SAN treated seeds. An irradiation with 5 min red light gave a germination of 98% both in water and in SAN. In water the effect of red irradiation could be reversed with a short irradiation (8 min) of far red light (17% germination), while in SAN solution the far red reversibility was poor (92% germination). If the far red light was given repeatedly (5 min per h) it had a slightly larger effect. If given continuously for 24 hours, the germination in water was decreased to 0.3% and in SAN solution to 9%. Possible mechanisms for the SAN effect are discussed.     

Effect of red radiation, kinetin and linuron on growth and ethylene production in Chlorella

Both short-term and continuous red radiation stimulated while far red radiation inhibited growth and ethylene production in Chlorella pyrenoidosa. Kinetin and linuron also affected culture density and ethylene production, depending on their concentration. Phytochrome might participated in the regulation of growth and ethylene production. This revised version was published online in July 2006 with corrections to the Cover Date.     

Effect of light intensity on algal biomass accumulation and biodiesel production for mixotrophic strains Chlorella kessleri and Chlorella protothecoide cultivated in highly concentrated municipal wastewater

In this research, the effect of light intensity on biomass accumulation, wastewater nutrient removal through algae cultivation, and biodiesel productivity was investigated with algae species Chlorella kessleri and Chlorella protothecoide. The light intensities studied were 0, 15, 30, 60, 120, and 200 μmol m(-2) s(-1). The results showed that light intensity had profound impact on tested responses for both strains, and the dependence of these responses on light intensity varied with different algae strains. For C. kessleri, the optimum light intensity was 120 μmol m(-2) S(-1) for all responses except for COD removal. For C. protothecoide, the optimum light intensity was 30 μmol m(-2) S(-1). The major components of the biodiesel produced from algae biomass were 16-C and 18-C FAME, and the highest biodiesel contents were 24.19% and 19.48% of dried biomass for C. kessleri and C. protothecoide, respectively. Both species were capable of wastewater nutrients removal under all lighting conditions with high removal efficiencies.     

Blue light induced inhibition of seed germination: The necessity of the fruit coats for the blue light response

The seeds (achenes) of Laportea bulbifera require a chilling to break their dormancy and are negatively photoblastic. Their germination is inhibited by both continuous blue light and continuous or prolonged far-red radiation. The germination of de-coated seeds, prepared by removing the fruit coats, however, was strongly inhibited by continuous far-red, but not by continuous blue light. Photoreversible germination by a brief irradiation with red light occurred when the chilled seeds were exposed to prolonged far-red light. These results suggest that far-red light may regulate the germination of L. bulbifera seeds through the phytochrome system which exists in the regions other than fruit coats and that the blue light reaction may be governed by other photoreceptor system(s).     

Influence of free Mg2+ on the kinetics of human erythrocyte phosphofructokinase

The influence of Mg2+ on the reaction catalyzed by human erythrocyte phosphofructokinase has been investigated using kinetic methods. The catalytic activity of PFK is dependent upon the presence of Mg2+ which constitutes with ATP the true Mg-ATP2- substrate. Free Mg2+ has no influence on the affinity of the enzyme for Mg-ATP2- substrate. Erythrocyte PFK is more inhibited by ATP4- and uncomplexed citrate than it is by Mg-ATP2- and Mg-citrate. Free Mg2+ relieves the MgATP2- and Mg-citrate inhibition under conditions where free ATP4-is negligible. We can assume that uncomplexed Mg2+ acts as positive effector by direct binding to the enzyme. These results emphasize the role of Mg2+ in the regulation of PFK activity in the erythrocyte.