Effects of blue light on respiration and carbon dioxide fixation in colorless Chlorella mutant cells

A colorless mutant of Chlorella vulgaris (Mutant #125) starvedin darkness, showed suppressed rates of respiration and darkCO₂ fixation, which were significantly recovered by illuminationwith blue light. The main CO₂ fixation product under blue lightwas aspartate. Such enhancements did not take place in cellsactively growing in the glucose medium. Both enhancing effectsof blue light (456 nm) were saturated at light intensities aslow as 400–800 erg.cm^-2.sec^-1. The action spectra forthese enhancing effects were similar to each other; both showedpeaks at 460 nm and 380 nm, which correspond to the absorptionmaxima of flavin. All these findings indicate that the samemechanism underlies the observed effects of blue light on CO₂fixation and respiration. The role of blue light which bringsabout the enhancements in CO₂ fixation and respiration is discussed. (Received June 1, 1974; )     

Negative Phototropism in Vaucheria terrestris Regulated by Calcium I. Dependence on Background Blue Light and External Calcium Concentration

In the presence of 1–8 mM Ca²⁺ unilateral 10–15min irradiation with blue light can elicit negative phototropicbending in the tip-growing coenocytic fresh water alga, Vaucheriaterrestris (Xanthophyceae), when it is simultaneously irradiatedwith strong blue or green background light. By changing wavelength,fluence rate, and duration of background light and holding thoseof unilateral light (456 nm, 1.7 Wm^–2) negative phototropicresponse was analyzed: the wavelength of the background lighthad to be shorter than 540 nm; red light (660 nm) was ineffectiveeven at very high fluence rates (100W.m^–2). The negativebending was strongly and specifically dependent on the externalCa²⁺ concentration. Other divalent cations, Mg²⁺ and Ba²⁺, wereeither toxic or quite ineffective; Sr²⁺ could partly supportthe growth, but mediated neither positive nor negative phototropicbending. The rate of tip-growth was not significantly alteredbetween 10⁶M and 10^-6 mM Ca²⁺. Pre-irrradiation with the backgroundlight slightly increased the negative curvature; whereas thatwithout subsequent simultaneous irradiation does not cause negativebending, but rather increases the positive curvature. Three-mindelayed start of background light did not cause negative bendingany more. The present results strongly suggest that blue light elicitsan influx of Ca²⁺ at the apex of Vaucheria and that the increasedcytoplasmic Ca²⁺ regulates the sensitivity and direction ofphototropic response. (Received June 23, 1988; Accepted September 7, 1988)     

Action of Near UV and Blue Light on the Photocontrol of Phycobiliprotein Formation; A Complementary Chromatic Adaptation

Action of near UV to blue light on photocontrol of phycoerythrin(PE) and phycocyanin (PC) formation was investigated with non-photobleachedTolypothrix tenuis and Fremyella diplosiphon; this study wasdone to evaluate the proposition of Haury and Bogorad [(1977)Plant Physiol., 60: 835] that near UV to blue light is as effectiveas green and red light for photocontrol of PE and PC formationin blue-green algae and that lack of the blue effect in previousexperiments was due to destruction of blue-absorbing pigment(s)by the photobleaching treatment involved in the experimentalmethod. In our present work, light effect was measured in heterotrophiccultures incubated in darkness following brief exposure to differentwavelengths of light. Results indicated that (1) near UV to blue light was not effectivefor induction of PE formation either in T. tenuis or in F. diplosiphon,and (2) PC formation was induced by near UV light at 360 nmbut not by blue light at 460 nm. These features are identicalwith those previously reported for photobleached cells but notwith those reported by Haury and Bogorad for non-photobleachedcells. We conclude that photobleaching treatment does not haveany influence on the action of near UV to blue light. Actionat 390 and 460 nm observed by Haury and Bogorad probably resultedfrom light effects other than photocontrol, e.g., the actionof photosynthesis. (Received December 18, 1981; Accepted April 8, 1982)     

Photochemical Apparatus Organization in the Diatom Cylindrotheca fusiformis: Photosystem Stoichiometry and Excitation Distribution in Cells Grown under High and Low Irradiance

The photochemical apparatus organization in the thylakoid membraneof the diatom Cylindrotheca fusiformis was investigated in cellsgrown under high and low irradiance. High light (HL, 200µE.m^–2.s^–1)grown cells displayed a relatively low fucoxanthin to chlorophyll(Chl) ratio, a low photosystem (PS) stoichiometry (PSII/PS I=1.3/1.0)and a smaller photosynthetic unit size in both PS I and PS II.Low light (LL, 30µE.m^–2.s^–1) grown cells displayeda 30% elevated fucoxanthin content, elevated PS II/PS I=3.9/1.0and larger photosynthetic unit size for PS II (a change of about100%) and for PS I (by about 30%). In agreement, SDS polyacrylamidegel electrophoresis of thylakoid membrane polypeptides showedgreater abundance of PS I, RuBP carboxylase and ATP synthasepolypeptides in HL cells. In contrast, LL grown cells exhibitedgreater abundance of light-harvesting complex polypeptides.Assuming an efficiency of red (670 nm) light utilization of1.0, the measured efficiency of blue (481 nm) light utilizationwas 0.64 (HL cells) and 0.72 (LL cells). The lower efficiencyof blue versus red light utilization is attributed to the quenchingof absorbed energy by non-fucoxanthin carotenoids. Differencesin the efficiency of blue light utilization between HL and LLgrown cells are attributed to the variable content of fucoxanthin.The results support the hypothesis of a variable Chl a-Chl c-fucoxanthinlight-harvesting antenna associated with PS II and PS I in Cylindrotheca. (Received February 10, 1988; Accepted April 6, 1988)     

Carbonic Anhydrase in Chlamydomonas reinhardtii II. Requirements for Carbonic Anhydrase Induction

Carbonic anhydrase (CA) activity in wild type cells of Chlamydomonasreinhardtii was low when cells were cultured under 2% CO₃ inthe light. When the gas phase was changed to air, CA activityincresaed as much as 20 fold over the next 24 hours. In contrast,CA activity did not change markedly in cells of the mutantspet 20-8 (PS II-negative), lip 10-2 (photophosphorylation-negative),and F60 (phosphoribulokinase-negative), when they were subjectedto the same induction regimen. DCMU (10^–5 M) and cydoheximide(3 µg/ml) severely inhibited the induction in wild typecells. No induction occured when CO₂ concentration was loweredin darkness. ³Present adress: Photoconversion Research Branch, Solar EnergyResearch Institute, Golden, Colorado 80401, USA. (Received June 7, 1982; Accepted December 25, 1982)     

Purification and Some Properties of Cytochrome b-560 from Nitrosomonas europaea

Cytochrome b-560 was purified to an electrophoretically homogeneousstate from Nitrosomonas europaea. It showed absorption peaksat 427, 530 and 560 nm in the reduced form. Its molecular weightwas estimated to be 44,000 by SDS-polyacrylamide gel electrophoresisand the same value was obtained on the basis of the contentsof haem and protein. The cytochrome was not autoxidizable anddid not react with CO. ¹Present address: Tokyo Research Center, TOSOH Corporation,Hayakawa, Ayase-shi, Kanagawa 252, Japan ²Present address: Faculty of Integrated Arts and Sciences, HiroshimaUniversity, Higashisenda-machi, Hiroshima 730, Japan (Received March 23, 1988; Accepted June 2, 1988)     

Effects of Light on Chlorophyll Formation in Cultured Tobacco Cells I. Chlorophyll Accumulation and Phototransformation of Protochlorophyll(ide) in Callus Cells under Blue and Red light

A marked accumulation of chlorophyll was observed in calluscells of Nicotiana glutinosa when they were grown under bluelight, while under strong red light no chlorophyll accumulated.This blue light effect saturated at an intensity of about 500mW.m^–2. The effects of white, blue and red light on the transformationof protochlorophyll (ide) (Pchl) accumulated in dark-grown calluscells were studied by following the changes in the intensityof fluorescence emitted by Pchl and different forms of chlorophyll(ide) (Chi). Pchl with a fluorescence maximum at 633 nm (absorptionmaximum: 630 nm) decreased slowly, concomitant with an increasein Chl having a fluorescence maximum at 677 nm (absorption maximum:675 nm), which was subsequently transformed, independently oflight, to Chi with a fluorescence maximum at 683 nm (absorptionmaximum: 680 nm). Both blue and red light of low intensitieswere effective for the phototransformation, while red light,but not blue light, of high intensities caused significant destructionof Pchl. An action spectrum for this photodestruction showedthat the maximum destruction took place at 630 nm. White lightof high intensities was effective for the photoreduction withonly slight destruction of Pchl, suggesting that blue lightcounteracts the destructive effect of red light. At low temperatures,however, blue light as well as red light of low intensitiescaused photodestruction of Pchl. It was inferred that blue lightenhances a certain step or steps involved in the productionof a reductant required for the photoreduction of Pchl to Chl. (Received July 3, 1981; Accepted November 11, 1981)     

Involvement of Calyculin A- and Okadaic Acid-sensitive Protein Phosphatase in the Blue Light Response of Stomatal Guard Cells

Calyculin A (CA) and okadaic acid (OA), inhibitors of proteinphosphatases, inhibited blue light (BL)-dependent H⁺pumpingin Vicia guard cell protoplasts at half-inhibitory concentrationsof 4.5 nM and 400 nM, respectively. Light-induced stomatal openingin Viciaepidermis was completely suppressed by CA at 100 nMand by OA at 1 µM. These results suggest that CA- andOA-sensitive protein phosphatase is involved in the BL responseof stomatal guard cells. (Received June 27, 1997; Accepted September 2, 1997)     

Spectral response of corn (Zea mays) in root geotropism

The primary roots of corn (Zea mays, Wisconsin hybrid 64A ?22R) show positive geotropism following exposure to light. Thisconfirms the works of other investigators. The curvature responsebegins at about 1 hr following irradiation and reached a plateauat 5 hr. A study of wavelengths 350–760 nm, using energiesof 2.24 ? 10¹⁴ photon cm^–2 and exposure times 60 sec,shows that the most effective light is at 660 nm with lessereffectiveness at 460 and 560 nm. The responses at 660 and 460nm are reversible by a far-red (730 nm) exposure, indicativeof the possible participation of phytochrome. Analyses of freshtips of corn roots with a dual-wavelength difference photometershow the phytochrome content in the root to be about 0.16 (OD) per gram fresh wieght. The requirement of light for thegeotropic growth response of corn roots might be an adaptivephenomenon. The occurrence of photomorphogenic activity in thegreen light should be of concern to those who use green as the"safe" light in "dark" experiments. ¹Work supported by U. S. NASA and U. S. ERDA. (Received September 26, 1977; )     

Action Spectrum in the Ultraviolet Region for Phototropism of Bryopsis Rhizoids

The phototropic response of the rhizoid of the marine coenocyticgreen alga Bryopsis plumosa to ultraviolet light (250–350nm) was investigated. The rhizoid exhibited negative bendingthat was due to bulging upon absorption of light in the UV region,as well as in the visible region, of the spectrum. The negativebending might not be a result of the inhibition of growth onthe irradiated side of the apical hemisphere by UV irradiationbecause growth inhibition was observed after bending had reacheda maximum within one to two hours. The action spectrum obtainedfrom fluence rate-response curves had a pronounced peak at 260nm and a small peak at 310 nm. The quantum effectiveness at260 nm was about five times that in the visible region. Phenylaceticacid (PAA), a potent inhibitor of flavin photoreactions, inhibitedthe phototropic response to both UV light and blue light withoutany obvious effect on tip growth. The inhibition of the phototropicresponse to blue light by PAA was partially overcome by rinsingthe alga with riboflavin-containing medium, which result suggeststhe involvement of flavins in the phototropism of Bryopsis rhizoids. (Received February 6, 1995; Accepted June 19, 1995)     

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)     

Wavelength effects on photosynthetic carbon metabolism in Chlorella

To study the wavelength-effect on photosynthetic carbon metabolism,¹⁴C-bicarbon-ate was added to Chlorella vulgaris 1 lh suspensionunder monochromatic blue (456 nm) and red (660 nm) light. Thelight intensities were so adjusted that the rates of ¹⁴CO₂ fixationunder blue and red light were practically equal. Analysis of¹⁴C-fixation products revealed that the rates of ¹⁴CO₂ incorporationinto sucrose and starch were greater under red light than underblue light, while blue light specifically enhanced ¹⁴CO₂ incorporationinto alanine, aspartate, glutamate, glutamine, malate, citrate,lipid fraction and alcohol-water insoluble non-carbohydratefraction. Pretreatment of the algal cells in phosphate mediumin the dark, which was essential for the blue light enhancementof PEP carboxylase activity, was not necessary to induce theabove wavelength effects. Superimposition of monochromatic bluelight at low intensity (450 erg.cm^–2.sec^–1) on thered light at saturating intensity caused a significant decreasein the rate of ¹⁴CO₂ incorporation into sucrose and increasein incorporation into alanine, lipid-fraction, aspartate andother related compounds, indicating that the path of carbonin photosynthesis is regulated by short wavelengdi light ofvery low intensity. Possible effects of wavelength regulationof photosynthetic carbon metabolism in algal cells are discussed. ¹ Part of this investigation was reported at the XII InternationalBotanical Congress, Leningrad, 1975 and the Japan-US CooperativeScience Seminar "Biological Solar Energy Conversion", Miami,1976. Requests for reprints should be addressed to S. Miyachi,Radioisotope Centre, University of Tokyo, Bunkyo-ku, Tokyo 113,Japan. ⁴ Present address: Department of Chemistry, Faculty of PharmaceuticalSciences, Teikyo Univ., Sagamiko, Kanagawa, Japan. (Received August 6, 1977; )     

Effcts of Blue Light and Ammonia on Nitrogen Metabolism in a Colorless Mutant of Chlorella

Illumination of a colorless mutant of Chlorella vulgaris 1lh(M125) with blue light enhanced both the uptake of nitrate andthe release of ammonia. These effects were not observed underillumination with red light. The release of ammonia was alsoenhanced by the addition of methionine sulphoximine (MSX), aninhibitor of glutamine synthetase (GS). Addition of MSX to culturesin the dark increased the rate of breakdown of starch. Algal cells grown in nitrate-containing medium did not showthe aminating activity of glutamate dehydrogenase (GDH). Additionof large (millimolar) amounts of ammonia in the dark resultedin the induction of NADPH-GDH activity and, in addition, a decreasein GS activity. From these results it appears that GS catalyzesthe primary step in the assimilation of ammonia in algal cellsgrown in nitrate-containing medium. Two isoforms (GS1 and GS2)of GS have been separated by ion exchange chromatography. Theactivities of both isoforms were decreased upon the additionof ammonia. Illumination of the alga with blue light at intensities up to10,000 mW m^–2 enhanced the measurable activity of GS invitro, while higher intensities were ineffective. In red lightno such effect was observed. The effects of blue light and ammonia on nitrogen metabolismin algal cells are discussed. (Received November 25, 1988; Accepted March 6, 1989)     

Effects of disalicylidenepropandiamine and near far-red light on 14CO2-fixation in Chlorella cells

1) With Chlorella ellipsoidea cells, in the presence of 5x10^–6M DSPD, or in its absence, the amounts of ¹⁴CO₂ incorporatedin P-esters, serine-plus-glycine and alanine were larger underred light than under blue light, whereas blue light specificallyincreased ¹⁴CO₂-incorporation in aspartate, glutamate, malateand fumarate (blue light effect). The amount of total ¹⁴C fixedunder blue or red light was greatly decreased by the additionof DSPD. When the concentration of DSPD was raised to 5x10^–4M, practically no radioactivity was found, under blue or redlight, in aspartate, glutamate and fumarate. Radioactivity inalanine was greatly increased. Effects of higher concentrationof DSPD are explained as due to the inhibition of PEP carboxylaseactivity in Chlorella cells. 2) The percentage incorporation of ¹⁴C into aspartate and theother compounds mentioned above, under near infra-red illuminationwas significantly smaller than that under blue light and wasalmost equal to that under red light. These results along withthe effect of 5x10^–6 M DSPD, exclude the possibility thatcyclic photophosphorylation is involved in the "blue light effect"mechanism. (Received December 12, 1969; )     

Studies on the Role of Photosynthesis in the Photoperiodic Induction of Flowering in the Short-Day Plants Kalanchoe blossfeldiana Poellniz and Xanthium pensylvanicum Wallr: I. THE REQUIREMENT FOR CO2 DURING PHOTOPERIODIC INDUCTION

It has been established that Kalanchoe blossfeldiana and Xanthiumpensylvanicum require CO₂ during the light period of short daysfor successful photoperiodic induction of flowering, even ifall but the induced leaf are held in normal air. In X. pensylvanicumfloral induction in normal air was independent of the starchstatus of the leaves but when reserves were reduced, lack ofCO₂ in the light suppressed floral induction to an even greaterextent. Injection into the induced leaf (Kalanchoe) or leaftip feeding (Xanthium) of carbohydrates, organic and amino acidsor several other metabolites failed to substitute for the CO₂requirement for induction. A small response was produced by10 mg ml^–1 sucrose in X. pensylvanicum while in normalair 25 parts 10^–6 ATP reduced the time to flowering inK. blossfeldiana and 10^–4 M proline was inhibitory. Anexperiment on the light requirement established a need for redlight ( max 660 nm) during photoperiods but red light alonedid not facilitate maximal induction. It is concluded that someearly, possibly labile, product of photosynthetic CO₂ fixationis essential to floral induction in these species.     

Intracellular Photoreceptive Site for Blue Light-Induced Cell Division in Protonemata of the Fern Adiantum--Further Analyses by Polarized Light Irradiation and Cell Centrifugation

The intracellular localization of the photoreceptive site forblue light-induced cell division in single-celled protonemataof Adiantum capillus-veneris L. was investigated using polarizedlight irradiation and protonemal cell centrifugation. The responseto irradiation with polarized blue light showed no dependenceon the direction of light polarization. However, centrifugationof the protonemata followed by microbeam irradiation showedthat the site of blue light perception could be displaced togetherwith the nucleus. Centrifugal treatment changed the distributionof intracellular organelles at the time of light exposure andbasipetally displaced the nucleus about 90µm. This treatmenthad no effect on the induction of cell division with blue lightif the protonemata were centrifuged again acropetally afterthe light treatment. Microbeam (30x30 µm²) irradiationwith blue light of the apical 45–75 ßm region,the receptive site of blue light in non-centrifuged cell, didnot induce cell division. However, cell division was inducedby irradiation of the nucleus-containing region, indicatingthat the photoreceptive site was displaced together with thenucleus by the centrifugation. These results suggest that theblue light receptor regulating cell division in Adiantum protonematais not likely to be located on the plasma membrane. (Received February 20, 1986; Accepted May 27, 1986)     

The Light-Growth Response of Vaucheria. A conditio sine qua non of the Phototropic Response?

This is the first report of a positive light-growth response(LGR) in tip-growing cells. Tip-growth of a coenocytic fresh-wateralga Vaucheria terrestris was temporarily accelerated by a shortpulse of blue light. The LGR occurred after a lag period shorterthan 1 min, and reached its maximum ca. 3 min after the onsetof blue light. The growth rate then rapidly decreased againand often showed a damped oscillation with a period of about10 min. If the blue light pulse was shorter than 2 min, themagnitude of the LGR seemed to obey the reciprocity law. Anothertype of growth promotion, the apical expansion, is brought aboutwhen the pulse is longer than 5 min. In this report, however,only the LGR which is caused by a short pulse of blue light,is dealt with. The threshold fluence at 456 nm was ca. 10 Jm^–2 at pH 7.0. The response was very sensitive to thepH of the medium: it was 5 J m ^–2 at pH 7.5, 150 J m^–2at pH 6, and 750 J m^–2 at pH 5. The phototropic responsealso showed a very similar pH-dependency between pH 5.5 andpH 6.5. The relationship between the positive LGR and positivephototropic response was found to be much closer in this tip-growingalga than in diffuse-growing cells. The possibility that theLGR is the primary and essential process preceeding the phototropicresponse is discussed.     

Effects of dark preincubation and chloramphenicol on blue light-induced CO2 incorporation in Chlorella cells

Chlorella cells incubated in the dark longer than 12 hr showedpronounced blue light-induced ¹⁴CO₂ fixation into aspartate,glutamate, malate and fumarate (blue light effect), whereasthose kept under continuous light showed only a slight bluelight effect, if any. 2) During dark incubation of Chlorellacells, phosphoenolpyruvate carboxylase activity and the capacityfor dark ¹⁴CO₂ fixation decreased significantly, whereas ribulose-1,5-diphosphatecarboxylase activity and the capacity for photosynthetic ¹⁴CO₂fixation (measured under illumination of white light at a highlight intensity) did not decrease. 3) In cells preincubatedin the dark, intracellular levels of phosphoenolpyruvate and3-phosphoglycerate determined during illumination with bluelight were practically equal to levels determined during illuminationwith red light. 4) The blue light effect was not observed incells incubated widi chloramphenicol, indicating that blue light-inducedprotein synthesis is involved in the mechanism of the effect. (Received April 9, 1971; )     

Red and blue light-stimulated proton efflux by epidermal leaf cells of the Argenteum mutant of Pisum sativum

Light stimulates leaf expansion in dicotyledons by increasingapoplastic acidification, cell wall loosening and solute accumulationfor turgor maintenance. Red and blue light enhance growth viadifferent photo-systems, but the cellular location and modesof action of these systems is not known. Here, the effect of red and blue light was studied on transportprocesses in epidermal cells of expanding leaves of the Argenteummutant of Pisum satlvum. Both red and blue light caused extraceiiuiaracidification by isolated epidermal tissue, which was stimulatedby extracellular K⁺ and inhibited by DCCD at 0.1 mol m^–3.Acidification induced by red compared with blue light showeddifferent saturating kinetics in fluence rate-response curves.Under near saturating light conditions the effects of red andblue light were additive. The red light-induced acidificationwas inhibited by far-red light while the blue light-inducedacidification was not. Light caused a hyperpoianzation of themembrane potential in epidermal strips, and stimulated ⁸⁶Rb⁺uptake by epidermal protoplasts. These results show that phytochromeand an additional blue light-photoreceptor function in isolatedepidermal cells to promote proton efflux, hyperpolarization,and cation uptake. Key words: Pisum sativum, light-induced acidification, ion transport, epidermis, photoreceptor