Electron Transport Coupled to Nitrate Reduction in a Photodenitrifier, Rhodopseudomonas sphaeroides forma sp. denitrificans

The electron transport system involved in nitrate reductionand its relationship to photosynthetic cyclic electron transportin a photodenitrifier, Rhodopseudomonas sphaeroides forma sp.denitrificans, were studied. Nitrate oxidized only b-type cytochromein the presence of cyanide, which inhibits nitrite reductase.Heptylhydroxyquinoline-N-oxide (HOQNO) inhibited the oxidationof b-type cytochrome by nitrate, but not the oxidation of b-and c-type cytochrome by nitrite. The inhibition by HOQNO wasovercome by phenazine methosulfate (PMS). Absorption changesof b-type cytochrome induced by illumination were in just theopposite directions for oxygen- and nitrate-oxidized cells;the cytochrome was reduced in oxygen-oxidized cells and oxidizedin nitrate-oxidized cells. Antimycin enhanced the reductionand inhibited the oxidation, but had no inhibitory effect onthe oxidation of b-type cytochrome by nitrate. Dithionite-reducedminus ferricyanide-oxidized difference spectra of cells at 77?Kshowed two b-type cytochrome components with a bands at 556.5and 562 nm. The proportion of the b-562 component decreasedin cells grown under denitrifying conditions. It was concludedthat a b-type cytochrome is involved in the nitrate reduction.The b-type cytochrome was presumed to be an alternative to thecytochrome b in the photosynthetic cyclic electron transport. ¹ Present address: Japanese Red Cross Tokyo-to Komagome BloodCenter, Komagome 2-2-2, Toshima-ku, Tokyo 170, Japan. (Received August 13, 1981; Accepted December 5, 1981)     

Cyclic strain and motion control produce opposite oxidative responses in two human endothelial cell types

The phenotype of endothelial cells (ECs) is specific to the vascular bed from which they originate. To examine how mechanical forces alter the phenotype of different ECs, we compared the effects of cyclic strain and motion control on reactive oxygen species (ROS) production and metabolism and cell adhesion molecule expression in human umbilical vein endothelial cells (HUVEC) vs. human aortic endothelial cells (HAEC). HUVEC and HAEC were subjected to cyclic strain (10% or 20%, 1 Hz), to a motion control that simulated fluid agitation over the cells without strain, or to static conditions for 24 h. We measured H₂O₂ production with dichlorodihydrofluorescein acetate and superoxide with dihydroethidium fluorescence changes; superoxide dismutase (SOD), catalase, and glutathione peroxidase (GPx) activities spectrophotometrically; and vascular cell adhesion molecule (VCAM)-1 and intercellular adhesion molecule (ICAM)-1 protein expression with Western blot analyses. HUVEC under cyclic strain showed 1) higher intracellular H₂O₂ levels, 2) increased SOD, catalase, and GPx activities, and 3) greater VCAM-1 and ICAM-1 protein expression, compared with motion control or static conditions. However, in HAEC, motion control induced higher levels of ROS, enzyme activities associated with ROS defense, and VCAM-1 and ICAM-1 expression than cyclic strain. The opposite responses obtained with these two human EC types may reflect their vessels of origin, in that HAEC are subjected to higher cyclic strain deformations in vivo than HUVEC. phenotype; reactive oxygen species; inflammation; shear stress     

The action of cyclic AMP on GA3 controlled responses VI. Characteristics of the promotion of light-inhibited seed germination in Phacelia tanacetifolia by GA3 and cyclic 3',5'-adenosine monophosphate

GA₃ and cyclic AMP were found to promote the germination (inlight) of light inhibited Phacelia tanacetifolia seed to thelevels obtained with dark controls. The GA₃ and cyclic AMP promotedgermination was inhibited by ABA and cycloheximide but not by6-methyl purine. The hormone and the nucleotide were not requiredfor the entire incubation period to obtain maximum germination. Theophylline and caffeine also promoted germination of theseseeds and 8-bromoadenosine cyclic AMP was found to be a betterpromoter of germination than cyclic AMP. Of a variety of nucleotidestested, only cyclic AMP promoted germination. This study supportsthe hypothesis that cyclic AMP is involved in some manner inGA₃-induced seed germination. (Received January 22, 1974; )     

SHORT COMMUNICATION Cyclic Nucleotides inFrankiaand Symbiotic Nodules

Cyclic adenosine 3',5'-monophosphate (cAMP) and cyclic guanosine3',5'-monophosphate (cGMP) contents of cultured cells ofFrankiastrainsoriginally isolated from nodules ofAlnus sieboldiana, MyricarubraandElaeagnus macrophyllawere measured by enzyme immunoassays(EIA).Frankiacells, cultured for 59–121 d, had cAMP contentsranging from 2.9 to 76.1 pmol mg^-1protein and cGMP contentsranging from 0.9 to 5.2 pmol mg^-1protein. FollowingFrankiaculture,the media contained extremely large quantities of cAMP and significantlevels of cGMP. The nature of accumulation and secretion ofcyclic nucleotides by slow-growingFrankiacells was comparableto that by a fast-growing actinomyceteStreptomyces lividansTK24,suggesting that secretion of cAMP byFrankiacells may occur throughthe cell membrane but not by cell lysis. cAMP and cGMP contentsin the symbiotic nodules, leaves and roots of actinorrhizalplants and leaves of non-actinorrhizal trees were also measured.The nodules of actinorrhizal woody plants(A. sieboldiana, E.macrophylla, E. umbellata, E. pungensandM. rubra)had cAMP contentsranging from 4 to 258 pmol g^-1f. wt and cGMP contents rangingfrom 1.1 to 5.2 pmol mg^-1protein. Most leaves and some rootsof actinorrhizal plants and all the leaves of non-actinorrhizalwoody plants examined contained small but significant amountsof cAMP and cGMP. This is the first report of significant contentsof cAMP and cGMP in culturedFrankiacells andFrankia-infectednodules. Possible roles of cyclic nucleotides as symbiotic signalsare discussed.Copyright 1998 Annals of Botany Company cAMP, cGMP, actinorrhizal plants, nodules,Frankia,symbiosis.     

Effects of Cyclic AMP on the Activity of Soluble Protein Kinases in Lemna paucicostata

Three protein kinases which phosphorylate histone were isolatedfrom cellular extract of Lemna plants. They were separated byelution from DEAE-Sephacel column and referred to as PI, PITand PHI. The PI protein kinase activity was partially inhibitedby 10µM cyclic AMP, cyclic GMP or cyclic IMP, while thePII enzyme was activated in the presence of these cyclic nucleotides.The PIII enzyme was cAMPindependent, but slightly inhibitedby cyclic CMP and cyclic UMP. The molecular weights of thesethree protein kinases were 165,000, 85,000 and 145,000, respectively,as estimated from Sephacryl S-300 gel filtration. A single cyclicAMP-binding protein was detected in the PII enzyme fractionby using the photoaffinity cAMP-analogue, 8-N₃-cAMP. The proteinwhich specifically bound [³H]-8-N₃-cAMP had an apparent molecularweight of 48,000 as determined by SDS-polyacrylamide gel electrophoresis.The phosphorylation of cellular proteins in Lemna was examinedby SDS-polyacrylamide gel electrophoresis. Four phosphorylatedpolypeptides were detected, the phosphorylations of which werestimulated by cAMP. The molecular weights of these four polypeptideswere 59,000, 19,000, 16,000 and 14,000, respectively. (Received January 26, 1983; Accepted April 13, 1983)     

NAD(P)H Dehydrogenase-Dependent Cyclic Electron Flow around Photosystem I in the Cyanobacterium Synechocystis PCC 6803: a Study of Dark-Starved Cells and Spheroplasts

Electron donation to P700⁺ through plastoquinone in the intersystemchain from both respiratory substrates and the photoreductantsin PSI has been shown to be mediated by the NAD(P)H-dehydrogenasecomplex (NDH) in Synechocystis PCC 6803 cells [Mi et al. (1992)Plant Cell Physiol. 33: 1233]. To confirm the participationof NDH in the cyclic electron flow around PSI, the redox kineticsof P700 and Chi fluorescence were analyzed in cells rendereddeficient in respiratory substrates by dark starvation and inspheroplasts. Dark-starved cells showed a high steady-state level of P700⁺under far-red (FR) illumination and the plastoquinone pool wasin a highly oxidized state. An NDH-defective mutant consistentlyshowed a high level of P700 oxidation under FR before and afterthe dark starvation. Donation of electrons either from exogenousNADPH or from photoreduced NADPH⁺ to the intersystem chain viaplastoquinone was demonstrated using spheroplasts from wild-typecells, but not those from the NDH-defective mutant, as monitoredby following changes in the kinetics of Chi fluorescence andthe redox state of P700. The electron flow to PSI via plastoquinone,mediated by NADPH, was sensitive to rotenone, Hg²⁺ ions and2-thenoyltrifluoroacetone, inhibitors of mitochondrial NDH andsuccinate dehydrogenase, but not to antimycin A. The pool sizeof electrons that can be donated to P700⁺ from the cytosol throughthe intersystem chain increased with increasing duration ofillumination time by actinic light and was sensitive to rotenonein both wild-type cells and spheroplasts, but no such resultswere obtained in the NDH-defective mutant of Synechocystis 6803.The results support our previous conclusion that NDH is a mediatorof both respiratory electron flow and cyclic electron flow aroundPSI to the intersystem chain in the cyanobacterium Synechocystis. (Received August 20, 1993; Accepted November 22, 1993)     

Infection of Coscinodiscus spp. by the parasitoid nanoflagellate Pirsonia diadema: II. Selective infection behaviour for host species and individual host cells

The parasitoid nanoflagellate (PNF) Pirsonia diadema is hostspecific for the marine centric diatom Coscinodiscus spp. Experimentsshowed that flagellates significantly prefer C. wailesii overC.granii as host species (interspecific selectivity). This preferencewas independent of light conditions (dark, irradiance of 10and 70 µmol m^–2 s^–1) and temperature (10 and15C). Among unicellular host diatoms, the infection behaviourwas selective for individual cells: already infected C.graniicells were more attractive for further flagellate attachmentthan non-infected cells (intraspecific selectivity). Individualcells (     

Ultrastructural Aspects of the Glandular Cells from the Secretory Trichomes and from the Cell Suspension Cultures of Achillea millefolium L. ssp. millefolium

The ultrastructure of the glandular cells of the floret secretorytrichomes from Achillea millefolium L. ssp. millefolium (yarrow)was examined before and after anthesis and compared with theultrastructure of the cells from the cell suspension culturesobtained from the same plant. The profuse tubular structuresobserved in the plastids of the glandular cells of the trichomesduring the pre-secretory stage were much reduced in the secretorystage and showed an osmiophilic content. Some endoplasmic reticulumprofiles could be seen adjacent to the plastids. Later in thesecretory stage, the secretion appeared in the periplasmic spacebetween the cells of the upper tiers and in the sub-cuticularspace. Finally the secretion was released by rupture of thecuticle. At the lag phase, the cells from the cell suspensioncultures of yarrow were characterized by the presence or plastidswith tubular structures, similar to those observed in the plastidsof the trichomes in the pre-secretory stage. By the end of thelag phase accumulations of starch were observed inside the plastids.At the beginning of the exponential phase, the tubular structuresof the plastids started to show an osmiophilic content and theaccumulations of starch were still present. At the end of thisphase starch disappeared from the plastids and only osmiophilictubular structures were observed. Rough endoplasmic reticulumas well as smooth endoplasmic reticulum profiles were frequentlyin close association with plastids and mitochondria. At thestationary phase a very large vacuole filled the cells, andin the remaining cytoplasm some endoplasmic reticulum profilesand osmiophilic droplets were observed.Copyright 1994, 1999Academic Press Achillea millefolium L. spp. millefolium, yarrow, ultrastructure, trichomes, glandular cells, plant cell suspension cultures     

The Effects of Light Irradiation on the Orientation of Microtubules in Seedlings of Avena sativa L. and Pisum sativum L.

The effects of light irradiation on the arrangement of corticalmicrotubules (MTs) were examined in etiolated A vena mesocotylsand coleoptiles, and in Pisum epicotyls. Elongation of A venamesocotyls ceased as a result of irradiation with white lightwithin 1 h. The predominant orientation of MTs became more longitudinalwithin 1 h in epidermal cells and changed from transverse tooblique, after the elongation ceased, in parenchymal cells.Irradiation with red and with blue light also caused cessationof cell elongation and the same changes in the orientation ofMTs. Elongation of Avena coleoptiles ceased as a result of irradiationwith white light within 24 h. The predominant orientation ofMTs became more longitudinal in epidermal cells and changedfrom transverse to oblique in parenchymal cells. The changein orientation of MTs in epidermal cells preceded that in parenchymalcells. In Pisum epicotyls, elongation ceased as a result ofirradiation with white light within 1 h. Although the orientationsof MTs in epidermal cells did not show any remarkable change,those in parenchymal cells changed from transverse to obliqueafter cell elongation ceased. The change in orientation of MTs and the cessation of cell elongationof A vena mesocotyls induced by white-light irradiation wereboth significantly retarded by treatment with IAA. This resultsuggests that IAA is involved in maintaining the transverseorientation of MTs in Avena mesocotyls. (Received February 22, 1989; Accepted August 2, 1989)     

The possible role of cyclic AMP in the control of genetic tumor induction

Young seedlings of the tumor-prone amphiploid Nicotiana suaveolensx N. langsdorffii, grown aseptically on nutrient medium, weretreated with 1x10^–2 M cyclic adenosine 3': 5'-monophosphate(cyclic AMP). The incidence of tumor formation was scored atregular intervals subsequent to exposure. Cyclic AMP causeda significant reduction in the rate of tumor formation. In addition,untreated plants grown on nutrient medium were harvested atregular intervals after the seeds had been sown. Cyclic AMPwas extracted, partially purified, and assayed radioimmunologically.The endogenous level of cyclic AMP in stem tissue was highestin young seedlings, rapidly fell reaching a low point in 18day-old plants, and eventually leveled off. The presence ofindoleacetic acid (IAA) in the growth medium at a final concentrationof 2x10^–5 M prevented the decline in cyclic AMP that occurredin seedlings grown on unsupplemented medium. (Received May 21, 1976; )     

Cyclic Changes in Transpiration of Sunflower Leaves in a Steady Environment

Concurrent cyclic changes of leaf temperature, transpirationrate, and stomatal aperture were found to occur in plants ofsunflower (Helianthus annuus L.) exposed to a steady aerialenvironment and with their roots in water. These cyclic changeswere less regular with plants in drying soils and did not occurat all with plants in soils with a water potential less than–120 J kg^–1. A sequence of possible events whichlead to the repeated cycling is suggested; these involve relativechanges in the turgor of epidermal and guard cells. The possibilitythat repeated cycling may be caused by a changing concentrationof carbon dioxide in the sub-stomatal cavities is also discussed.The irregularity and eventual cessation of these cyclic changesare considered to be due to the reduction in turgor of the epidermaland guard cells as the availability of water to these cellsis reduced.     

Photoinhibition and light-induced cyclic electron transport in ndhB(-) and psaE(-) mutants of Synechocystis sp. PCC 6803

The ndhB^– and psaE^– mutants of the cyanobacteriumSynechocystis sp. PCC 6803 are partly deficient in PSI-drivencyclic electron transport. We compared photoinhibition in thesemutants to the wild type to test the hypothesis that PSI cyclicelectron transport protects against photoinhibition. Photoinhibitorytreatment greatly accelerated PSI cyclic electron transportin the wild type and also in both the mutants. The psaE^–mutant showed rates of PSI cyclic electron transport similarto the wild type under all conditions tested. The ndhB^–mutant showed much lower rates of PSI cyclic electron transportthan the wild type following brief dark adaptation but exceededwild type rates after exposure to photoinhibitory light. Thewild type and both mutants showed similar rates of photoinhibitiondamage and photoinhibition repair at PSII. Photoinhibition atPSI was much slower than at PSII and was also similar betweenthe wild type and both mutants, despite the known instabilityof PSI in the psaE^– mutant. We conclude that photoinhibitorylight induces sufficient PSI-driven cyclic electron transportin both the ndhB^– and psaE^– mutants to fulfill anyrole that cyclic electron transport plays in protection againstphotoinhibition. ⁴ Corresponding author: E-mail, sherbert@uwyo.edu; Fax, +1-307-766-2851;Phone, +1-307-766-4353.     

Light-induced reactions of ubiquinone in photosynthetic bacterium, Chromatium D. II. Effects of inhibitors and other experimental conditions

Light-induced reactions of ubiquinone in starved cells of ChromatiumD, were investigated under various experimental conditions. The "low amplitude" photoreduction observed on illuminationwithout addition of substrate under aerobic conditions and the"light-off spikes" observed under anaerobic conditions wereshown to be related to the cyclic flow of electrons in the photosyntheticsystem. Both of these were inhibited by o-phenanthroline, HOQNOand piericidin A. The "high amplitude" photoreduction of ubiquinoneobserved in the presence of thiosulfate under aerobic conditionswas inhibited by PCMB, PMA and KCN. The "high amplitude" photooxidationof ubiquinone in the presence of malate under anaerobic conditionswas inhibited by HOQNO, piericidin A, rotenone and PMA. In the presence of KCN and succinate, similar "high amplitude"photooxidation was obtained even under aerobic conditions. Underaerobic conditions, the addition of malate did not affect the"high amplitude" photoreduction due to thiosulfate. On the contrary,under anaerobic conditions, the "high amplitude" photooxidationwith malate was almost completely abolished by the additionof thiosulfate. This apparent counteraction of the oxidationand reduction of ubiquinone was inferred to represent an optimumstate of its efficient turn-over in the noncyclic electron transportchain of photosynthesis inChromatium. ¹This article is the second of a series of studies previouslyreported under the same title in Vol. 8 of this journal (Reference14). (Received August 23, 1968; )     

Involvement of Cyclic AMP in Adventitious Bud Initiation of Torenia Stem Segments

Adventitous bud initiation in epidermis of Torenia stem segmentscultured in vitro, which is usually induced by cytokinin, couldbe induced by application of dibutyryl cyclic AMP in the absenceof cytokinin. Similar stimulatory effects on bud initiationwere observed when various promoting reagents to accumulateendogenous cyclic AMP were added, such as activators for adenylatecyclase, forskolin and prostaglandin E₁, and inhibitors of cyclicnucleotide phospho-diesterase, theophylline and isobutyl methylxanthine. Endogenous content of cyclic AMP in Torenia stem segmentswere increased by the application of the above chemicals, calciumionophore or cytokinin. These results suggested that endogenousconcentrations of cyclic AMP were involved in adventitious budinitiation of Torenia stem segments. Furthermore, some inhibitorsof protein kinases inhibited bud initiation induced by cyclicAMP-accumulating reagents. (Received August 15, 1989; Accepted November 4, 1989)     

Chromatic Regulation of Photosystem Composition in the Photosynthetic System of Red and Blue-Green Algae

A chromatic adaptation in the photosynthetic quantum yield forthe light mainly absorbed by chlorophyll a (Chl a light) firstfound by Yocum (1951) was studied with one red and three blue-greenalgal strains. When the cells were grown under a weak Chl alight, the quantum yield in all the strains increased. Comparisonof photosystem (PS) compositions, including phycobilin (PBP)and Chl a antennae, reaction centers I and II, in the cellsgrown under the light mainly absorbed by PBP and Chl a revealedthat changes in quantum yield could be attributed to changesin the ratio of PS I/II; PS I/II becomes larger than 1 underPBP light but decreases to 1 in most cases under Chl a light.The change in the PS I/II ratio is due solely to the changesin the PS I population in the cell; PS II remains constant.These results are similar to the intensity-dependent responsein PS composition. A common hypothesis for both the chromatic and intensity-inducedregulation of PS composition was proposed based on the ideaof balance between the electron flow from H₂O to NADP drivenby PS I and II and the cyclic one driven by PS I. (Received May 16, 1985; Accepted September 4, 1985)     

Mode of Action of the Crustacean Hyperglycemic Hormone

Glucose levels in crayfish hemolymph are enhanced by the crustaceanhyperglycemic hormone (CHH); at present there is some evidencethat this action is mediated by cyclic nucleotides. CHH is capableof stimulating adenylate cyclase in the abdominal muscle. Thereis an increase in cyclic AMP and cyclic GMP contents in hepatopancreasand abdominal muscle after CHH injection. Cyclic nucleotidesare able to evoke the same reaction as CHH in vivo and in vitro.Cyclic nucleotide-dependent protein kinases are activated bythe hormone, which leads to a phosphorylation and thereforeinhibition of glycogen synthase. So far, an effect of purifiedhormone on phosphorylase and phosphorylase kinase has not beendemonstrated in the abdominal muscle.     

Cyclic Nucleotides and In Vitro Plant Cultures: I. INDUCTION OF ORGANOGENESIS IN TOBACCO (NICOTIANA TABACUM CALLUS CULTURES

Mangat, B. S. and Janjua, S. 1987. Cyclic nucleotides and invitro plant cultures. I. Induction of organogenesis in tobacco(Nicotiana tabacum) callus cultures.—J. exp. Bot. 38:2059–2067. The possibility that cyclic nucleotides have a mediatory rolesimilar to cytokinins in plant tissue cultures was examined.Calli obtained from tobacco pith tissue were incubated on growthmedia supplemented with either cyclic AMP, cyclic GMP, adenosineor guanosine, in concentrations ranging from (mg dm^–3)0 to 2·0 together with 2·0 mg dm^–3 of IAA.Results were compared with identical calli grown on media containingcomparable amounts of kinetin and IAA. Increase in callus growthwas observed on all media containing cyclic AMP, cyclic GMP,adenosine, guanosine or kinetin. Adenosine or guanosine didnot promote organogenesis. Low concentrations (0·02 and0·05 mg dm^–3) of kinetin stimulated extensive rootdevelopment. Some root formation was also elicited with higheramounts of cyclic AMP (0·1 and 0·2 mg dm^–3)or cyclic GMP (0·2 and 0·5 mg dm^–3). Bothkinetin and cyclic GMP promoted shoot differentiation. However,in contrast to kinetin, cyclic GMP induced organogenesis atlower concentrations (0·02 and 0·1 mg dm^–3).The addition of 2·0 mg dm ^–3 of cyclic AM P toIAA-free growth media elicited shoot differentiation. This wasalso the case with a similar concentration of kinetin or cyclicGMP. Results suggest cytokinin activity for the two cyclic nucleotides. Key words: Tobacco, Nicotiana tabacum, tissue culture, cyclic nucleotides, cyclic AMP, cyclic GMP organogenesis     

Stomata on the Primary Root of Pisum sativum L.

The first record of stomata on a non-specialized root was obtainedby scanning electron microscopy of 4-d-old Pisum sativum L.In some cases subsidiary cells were trichoblasts. Stomata andthe root triarch vascular structure were simultaneously presentin transverse sections through the root. Pisum sativum, pea, root stomata, guard cells, trichoblasts     

Effects of gibberellic acid and cyclic 3',5'-adenosine monophosphate on the flowering of Eschscholtzia californica Cham., a qualitative long day plant.

GA₃ and cyclic AMP induce floral buds in Eschscholtzia californicaunder noninductive short days and hasten it under inductivelong day conditions. (Received June 8, 1976; )