Effects of Blue Light on the Uptake of Ammonia and Nitrate by a Colorless Mutant of Chlorella

A colorless mutant of Chlorella kessleri, grown in darknessin a medium that contained nitrate and glucose, took up ammoniamore efficiently than nitrate. Irradiation with blue light inhibitedthe uptake of ammonia but, conversely, the uptake of nitratewas enhanced by blue light. These effects were not observedunder illumination with red or far-red light. The inhibitionby blue light of the uptake of ammonia was abolished in thepresence of nitrate. The charge compensation for the uptakeof ammonia was achieved by the immediate release of potassiumions and this release was followed by release of protons, therate of the latter process being strongly reduced by blue light. The effects of blue light on the uptake of ammonia and nitratein algal cells are discussed. (Received July 28, 1994; Accepted January 30, 1995)     

Effects of Light on Growth and Glucose Consumption in Colorless Chlorella Mutant Cells

Light inhibited the growth and glucose consumption of colorlessmutant cells of Chlorella vulgaris (# 125). Sugar consumptionwas also inhibited in a medium containing a hexose such as D-fructose,D-galactose and D-mannose. Blue light strongly inhibited growth and glucose consumptionbut red light only slightly affected them. Respiration was notinhibited by blue light. The inhibitions of growth and glucoseconsumption were saturated at light intensities as low as 800mW.m^–2 and continued in the dark for at least one dayafter brief illumination with white light. The half-maximumeffect was observed with 15 min of illumination in both casesand the action spectra for light-induced inhibitions of growthand glucose consumption were similar, both showing peaks at370, 457 and 640 nm. The role of light in the inhibitions of growth and glucose consumptionis discussed. (Received June 18, 1984; Accepted October 29, 1984)     

Some Biochemical Changes Related to Starch Breakdown Induced by Blue Light Illumination and by Addition of Ammonia to Chlorella Cells

Illumination with blue light enhanced the production of ammoniaby cells of C. vulgaris 11h, while no such effect was inducedby red light illumination. Addition of ammonia caused increasesin ATP levels and decreases in Pi and ADP levels. When 5 mMNH₄Cl was added to phosphorylase and amylase isolated from thecells of C. vulgaris 11h, their activities increased about 5–15%and 40–100%, respectively. (Received June 21, 1986; Accepted December 23, 1986)     

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 Light-Induced In Vivo Absorbance Changes and In Vitro Activation of Nitrate Reductase in a Nitrate-Starved Chlorella Mutant

Illuminating a colorless mutant of Chlorella vulgaris 11h (M125)with blue light caused a reversible photoreduction of b-typecytochrome, i.e., absorbance increases at 423, 525 and 557 nm.This light-induced reduction of cytochrome b was most pronouncedin nitrate-starved cells, which showed some blue light responsesin carbon metabolism, including enhancement of respiration byblue light as reported previously. Prolonged illumination withblue light caused a decrease in the rate of the reduction. The photoactivation of nitrate reductase in the mutant cellswas studied in both cell-free crude extract and purified enzyme.The absorption spectrum of purified enzyme showed three peaksat 423, 525 and 557 nm after the addition of a reductant, indicatingthat the spectrum is that of cytochrome b associated with nitratereductase. Nitrate reductase activity was easily enhanced byblue light illumination after 1 min; red light had no effecton it. The blue light activation of nitrate reductase was notsignificant in growing cells, which showed its high activity. The relationship between the blue light-induced reduction ofcytochrome b and carbon metabolism is discussed. (Received September 30, 1987; Accepted February 9, 1988)     

Effect of Light Quality on Nitrogen Metabolism of Radish Plants

The long-term action of blue or red light on nitrogen metabolism was studied in radish (Raphanus sativus L.) plants. The potential activity of nitrate reductase (NR) in vivo and its maximum activity in vitro, the content of soluble protein and free amino acids were determined in the course of the growth of a third leaf of radish plants. The effect of light quality on NR activity was found to depend significantly on the stage of leaf development. Blue light (BL) stimulated NR activity in leaves, when their areas were about 11–13% of the fully developed leaves. The efficiency of red light (RL) was significantly lower, because the maximum NR activity was observed in the leaves developed to the stage, when their areas were 38–40% of the final one. The comparative analysis of the pool of free amino acids in expanding leaves of BL- or RL-grown plants revealed significant changes in the contents of individual amino acids. Despite a higher accumulation of two amino acids in the leaves of BL-grown plants, namely, Asp (27% as compared to 13–16% in the RL-grown leaves) and Gly (5% against 2.5% in RL-grown leaves), the BL-grown leaves also demonstrated a significant decrease in Ala (10% as compared to 23% in the RL-grown leaves) and some decrease in the amounts of Ser and Gly. The content of soluble protein in a juvenile BL-grown leaf was observed to decrease gradually during leaf development. However, the protein content in the BL-grown leaf was always higher than in the RL-grown leaf of the same age. We concluded that the photoregulatory action of BL on NR activity determined the different rates of nitrogen assimilation in BL- and RL-grown plants.__________Translated from Fiziologiya Rastenii, Vol. 52, No. 3, 2005, pp. 349–356.Original Russian Text Copyright © 2005 by Maevskaya, Bukhov.     

Adaptation of the Photosynthetic Apparatus of Chlorella and Ankistrodesmus to Blue and Red Light

The mode of adaptation of the photosynthetic apparatus of three unicellular green algae, Ankistrodesmus braunii, Chlorella fusca and Chlorella saccharophila to red and blue light are documented by the fluence-rate curves of photosynthetic oxygen evolution. For all three algae tested photosynthetic capacity, respiration and light compensation point were higher for cells grown under red light, while the chlorophyll content increased in blue light-grown cells. Blue light-adapted cells have a lower chlorophyll a to chlorophyll b ratio and more chlorophyll in the light-harvesting system than red light-adapted cells, as shown in the electrophoretic profile of the pigment-protein complexes. It is concluded that the action of red light resembles that of high levels of white light, while blue light causes the same effects as low levels of white light. In agreement with previous publications these findings indicate that the mode of adaptation to different light qualities is ubiquitous in unicellular green algae.     

Ammonia Regulation of Intermediary Metabolism in Photosynthesizing and Respiring Chlorella pyrenoidosa: Comparative Effects of Methylamine

The natural ¹³C abundance (¹³C value) of the field-grown leguminousplants (soybean, kidney bean, pea, azuki bean, mung bean, peanutand cowpea) was investigated by mass spectrometry with a precisionbetter than %0.2 for ¹³C. Among organs of premature plants,the leaves had the most negative values, and the nodules generallyhad the least negative values, and other organs, fruits, stemsand roots, showed intermediate values. In the soybeans so farinvestigated, the grains of nodulating plants exhibited higher¹³C values than nonnodulating lines. The ¹³C values of the grainsvaried depending on the species: peanuts showed the most negativevalues. Possible causes underlying these variations are discussed. (Received March 2, 1983; Accepted May 27, 1983)     

Regulation of Glutamine Synthetase by Light and during Nitrogen Deficiency in Synchronous Chlorella sorokiniana

A method is described to achieve density labeling of proteins in unicellular algae by using ¹³CO₂. This is a satisfactory procedure especially for work on nitrogen metabolism. The increase in activity of glutamine synthetase (EC 6.3.1.2.) and glutamate synthase (EC 1.4.7.1.) in Chlorella sorokiniana mediated by a dark/light shift and by nitrogen starvation were investigated. Using the method of density labeling and isopycnic centrifugation, we demonstrated that the increase in enzyme activity after a dark/light shift is based on activation rather than de novo synthesis. The increase in enzyme activity after transfer to nitrogen-deficient medium is based both on activation and de novo synthesis.     

Light-Induced Changes in the Molecular Size of Starch in Colorless Chlorella Cells

Starch from colorless mutant cells of Chlorella vulgaris (#125), grown heterotrophically in the dark, was fractionatedby agarose gel chromatography. The molecular weight distributionof starch showed only one pronounced maximum at 2?10⁶ (L-starch).Exposure of cells to white light decreased significantly inthe amounts of the total starch and L-starch. This also wastrue under non-growth conditions, the same effect being foundto depend on blue light. The role of light on starch degradationis discussed. (Received November 29, 1984; Accepted February 25, 1985)     

Light Inhibits Flagellation in a Chlamydomonas Mutant

We have isolated a new flagellar mutant in Chlamydomonas reinhardtii.When the mutant was cultured under the white fluorescent lamp({small tilde}4,800 lux), most cells had no flagella. However,when the cultures were put in the dark, flagellation occurred.Greater than 70% of the cells had flagella within 12–16h after the transfer. The flagellar morphology varied from "rod-shape"(same as the wild-type flagella) to "disk-shape". The disk-shapedflagella had the axonemes which were curved into a loop withinthe swollen membrane. Hence, this mutant is called loop-1. Light-inhibitionof flagellation was restored in the presence of 10^–5 MDCMU. The spectral dependency of the photo-inhibition of flagellation,determined using the Okazaki Large Spectrograph, showed maximaleffectiveness at 400–420 nm and 600–680 nm. Theseresults suggest that photosynthesis inhibits flagellation ofloop-1 cells. (Received July 27, 1989; Accepted January 29, 1990)     

Regulation of Carbohydrate Breakdown of Chlorella Mutant No. 20, Studied by 31P NMR Spectroscopy and Enzymatic Analysis: 1. Regulation under Blue Light Conditions

The regulation of carbohydrate degradation, especially of itsglycolytic part, is studied in darkness and under blue lightconditions in the chlorophyll-free Chlorella mutant no. 20.The mass action ratios of glycolytic reactions are calculatedfrom the concentrations of the respective intermediates, measuredvia enzymatic analysis and ³¹P NMR spectroscopy. Comparisonwith the equilibrium constants indicates that the reactionscatalyzed by hexokinase, phosphofructokinase, and pyruvate kinaseare able to control the glycolytic pathway. Blue light illumination leads to rapid changes in the levelof various metabolites. The decrease in PEP proves the regulatoryrole of pyruvate kinase in vivo, a finding supported by thecrossover plot as well as by the move of the mass action ratiotowards equilibrium. Also, the starch degrading enzymes phosphorylaseand possibly amylase, but not phosphofructokinase, seem to beinvolved in the regulation of carbohydrate breakdown in bluelight. The kinetics of the different blue light responses clearly indicatethat not the increase in respiratory O₂ uptake, but an appropriatechange in the concentration of effectors of the regulatory enzymespyruvate kinase, phosphorylase, and maybe amylase is a primaryblue light effect in the Chlorella mutant. The involvement ofa proton pump seems to be ruled out by the constant intracellularpH measured by in vivo ³¹P NMR spectroscopy. (Received October 5, 1987; Accepted January 7, 1988)     

Genetic Characterization of the Carotenoid Biosynthetic Pathway in Methylobacterium extorquens AM1 and Isolation of a Colorless Mutant

Genomic searches were used to reconstruct the putative carotenoid biosynthesis pathway in the pink-pigmented facultative methylotroph Methylobacterium extorquens AM1. Four genes for putative phytoene desaturases were identified. A colorless mutant was obtained by transposon mutagenesis, and the insertion was shown to be in one of the putative phytoene desaturase genes. Mutations in the other three did not affect color. The tetracycline marker was removed from the original transposon mutant, resulting in a pigment-free strain with wild-type growth properties useful as a tool for future experiments.     

蓝光对绿豆下胚轴愈伤组织形成和生长过程中蛋白质代谢的影响

蓝光、白光和黑暗对绿豆下胚轴愈伤组织形成和生长过程中蛋白质代谢的影响不同。培养后３～１８ ｄ ,蓝光处理材料的可溶性蛋白质含量明显高于白光处理,更高于黑暗培养的材料。蓝光和白光明显促进３Ｈ亮氨酸掺入蛋白质,而蓝光和白光处理后游离氨基酸含量与黑暗对照相比,下降时间早,幅度大。在培养过程中,蛋白酶活性的变化与游离氨基酸相似。蛋白质合成抑制剂环己酰亚胺（ＣＨＭ） 抑制愈伤组织生长,其中以蓝光最大,白光次之,黑暗最小。在培养基中加入ＣＨＭ 愈早,抑制程度愈大。实验表明,ＣＨＭ 抑制愈伤组织蛋白质合成,也是以蓝光最甚。由此可见,蓝光促进绿豆下胚轴愈伤组织的形成、生长和蛋白质合成。     

Differential Regulation of Nitrogen Metabolism in the Wild Type and the High-Light-Tolerant Mutant Cells of Anacystis nidulans

The glutamine synthetase activity in the wild type and high-light-tolerant mutant of Anacystis exhibited differential response to the increasing light intensity (2–40 W/m²). As evident from the results, the glutamine synthetase (GS) activity in the wild type is more dependent on respiration, whereas the GS enzyme in the mutant cells derived its carbon and energy from photosynthesis. Further, results revealed that the reduced GS activity in the wild-type cells under the high-light stress was accompanied by high aspartate amino transferase (AST/GOT) activity and low alanine amino transferase (ALT/GPT) activity. On the contrary, high GS activity in the mutant cells was accompanied by low AST/GOT enzyme activity and high ALT/GPT activity. It was inferred that mutant and wild-type cells adapt to the high-light stress by different mechanisms.     

Nitrogen Metabolism in Plant Cell Suspension Cultures: II. Role of Organic Acids during Growth on Ammonia

Tobacco cells (Nicotiana tabacum) are capable of growth on ammonia as a sole nitrogen source only when succinate, malate, fumarate, citrate, α-ketoglutarate, glutamate, or pyruvate is added to the growth medium. A ratio between the molar concentrations of ammonia to succinate (as a complementary organic acid) in the growth medium of 1.5 was optimal. Succinate had no effect on the rate of uptake of ammonia from the medium into the cells although it did affect the intracellular concentration of ammonia. However, the changes were not sufficient to explain inhibition of growth as being due to ammonia toxicity. The radioactivity from ¹⁴C-succinate was incorporated into malate, glutamate, and aspartate within 2 minutes.     

Glucose and Fructose Metabolism in a Phosphoglucoisomeraseless Mutant of Saccharomyces cerevisiae

A mutant of Saccharomyces cerevisiae deficient in phosphoglucoisomerase (EC 5.3.1.9) is described. It does not grow on glucose or sucrose but does grow on galactose or maltose. Addition of glucose to cultures growing on fructose, mannose, or acetate arrests further growth without altering viability; removal of glucose permits resumption of growth. Glucose causes accumulation of nearly 30 mumoles of glucose-6-phosphate per g (wet weight) of cells and suppresses synthesis of ribonucleic acid. Inhibition of growth by glucose does not appear to be due to a loss of adenosine triphosphate or inorganic orthophosphate. The mutant, however, utilizes glucose-6-phosphate produced intracellularly. Release of carbon dioxide from specifically labeled glucose suggests a C-l preferential cleavage. The kinetics of glucose-6-phosphate accumulation during glucose utilization in the mutant is not consistent with the notion that the utilization of glucose is controlled by glucose-6-phosphate.