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1.
Transient time courses ("induction") of the intensity of milliseconddelayed fluorescence, measured between 0.1 and 3.9 msec after0.9 msec-light excitation period, were studied and comparedwith the induction of the prompt fluorescence in uncoupled spinachchloroplasts at temperatures between 20 and –170°C. The intensity of delayed fluorescence decreased during illumination,whereas that of prompt fluorescence increased at temperatureslower than 0°C. This type of correlation between these twotime courses also was observed in the presence of 3-(3',4'-dichlorophenyl)-l,l-dimethylurea,ferricyanide or dephenylcarbazide at various temperatures. Thecorrelation could be explained in terms of the redox statesof the primary electron donor and the acceptor molecules inphotosystem II. The enhancement of the intensity of the delayed fluorescenceobserved on cooling the material to –50°C was dueto the inhibition of electron donation from the water-splittingsystem by cooling, because enhancement was eliminated by anaddition of diphenylcarbazide. Below –50°C the intensity of delayed fluorescencedecreased on cooling and became too low to detect below –170°Cunder any of the conditions tested. This result is discussedin relation to the energy levels of redox components in reactioncenter. (Received May 27, 1980; )  相似文献   

2.
Dark-adapted intact spinach chloroplasts exhibited two peaks,P and M1, at the early phase of fluorescence induction and atransient reduction of cytochrome f shortly after its initialphotooxidation and in parallel to the appearance of P. Analysisof the peak P and the transient reduction of cytochrome f indicatedthat electron transport in intact spinach chloroplasts was regulatedby light: electron transport was inactivated at the reducingside of photosystem I in the dark-adapted chloroplasts but rapidlyreactivated by illumination. The fluorescence peak M1 was correlatedto the proton gradient formed across the thylakoid membrane. Effects on P and transient reduction of cytochromef of NO2,3-phosphoglycerate (PGA) and oxalacetate (OAA), which can penetrateinto intact chloroplasts and accept electrons at different sitesafter photosystem I, were studied to determine the site of thelight regulation. NC2, which receives electrons fromreduced ferredoxin, markedly diminished both P and the transientreduction of cytochrome.f, whereas PGA and OAA, the reductionsof which are NADP-dependent, failed to affect the two transients.The ineffectiveness of PGA and OAA could not be attributed tothe dark inactivation of glyceraldehyde-3-phosphate and malicdehydrogenases, because dark-adapted chloroplasts still retainedsufficiently high levels of the enzyme activities. The resultsindicate that electron transport in intact spinach chloroplastsis regulated by light after ferredoxin but before NADP, i.e.,at the reducing terminal of the electron transport chain. (Received May 29, 1980; )  相似文献   

3.
Leaves of Zea mays L. cv. LG11 were chilled for 6 h at 5 °Cin a high photon flux density. On return to 20 °C, the leavesshowed a 45% decrease in the apparent quantum yield of photosyntheticoxygen evolution. The effects of this chill-treatment on thechlorophyll fluorescence induction kinetics of the leaves indicateda 20–25% decrease in the primary photochemical quantumyield of photosystem II. The fluorescence emission spectra ofthese leaves demonstrated a marked modification in the distributionof excitation energy within the photochemical apparatus of thethylakoid membranes, such that photosystem I was excessivelyfavoured with respect to photosystem II. These chill-inducedchanges would result in an enhancement of cyclic over non-cyclicelectron transport and account for a decrease in the apparentquantum yield of photosynthetic oxygen evolution. Key words: Zea mays, Chilling, Photosynthesis, Thylakoids  相似文献   

4.
In this study, we evaluated how cadmium inhibitory effect on photosystem II and I electron transport may affect light energy conversion into electron transport by photosystem II. To induce cadmium effect on the photosynthetic apparatus, we exposed Chlamydomonas reinhardtii 24 h to 0–4.62 μM Cd2+. By evaluating the half time of fluorescence transients O–J–I–P at different temperatures (20–30°C), we were able to determine the photosystem II apparent activation energies for different reduction steps of photosystem II, indicated by the O–J–I–P fluorescence transients. The decrease of the apparent activation energies for PSII electron transport was found to be strongly related to the cadmium-induced inhibition of photosynthetic electron transport. We found a strong correlation between the photosystem II apparent activation energies and photosystem II oxygen evolution rate and photosystem I activity. Different levels of cadmium inhibition at photosystem II water-splitting system and photosystem I activity showed that photosystem II apparent activation energies are strongly dependent to photosystem II donor and acceptor sides. Therefore, the oxido-reduction state of whole photosystem II and I electron transport chain affects the conversion of light energy from antenna complex to photosystem II electron transport.  相似文献   

5.
Preincubation of chloroplasts from pea leaves (Pisum sativum L. cv. Kelvedon) with 0.5 millimolar ferricyanide in the dark, caused a parallel inhibition of the rate of rise of the variable fluorescence and the rate of electron transport. Both reactions were inhibited to a similar extent by varying the time of preincubation, the concentration of ferricyanide during preincubation, and by raising the concentration of salts in the preincubation medium. Ferricyanide treatment of Tris-washed chloroplasts did not inhibit electron transport from the Photosystem II (PSII) electron donor 1,5-diphenylcarbazide to methylviologen. The inhibition of the variable fluorescence rise and of NADP reduction (caused by ferricyanide pretreatment) was bypassed by addition of the PSII electron donor couple hydroquinone/ascorbate. It was concluded that preincubation of chloroplasts with ferricyanide in the dark inhibited electron transport between water and PSII.  相似文献   

6.
Photosynthetic Properties of Guard Cell Protoplasts from Vicia faba L.   总被引:3,自引:0,他引:3  
Guard cell protoplasts were isolated enzymatically from theepidermis of Vicia faba L. and their photosynthetic activitieswere investigated. Time courses of light-induced changes inthe chlorophyll a fluorescence intensity of these protoplastsshowed essentially the same induction kinetics as found formesophyll protoplasts of Vicia. The transient change in thefluorescence intensity was affected by DCMU, an inhibitor ofphotosystem II; by phenylmercuric acetate, an inhibitor of ferredoxinand ferredoxin NADP reductase; and by methyl viologen, an acceptorof photosystem I. Low temperature (77 K) emission spectra ofthe protoplasts had peaks at 684 and 735 nm and a shoulder near695 nm. A high O2 uptake (175 µmol mg–1 Chl hr–1)was observed in guard cell protoplasts kept in darkness, whichwas inhibited by 2 mM KCN or NaN3 by about 60%. On illumination,this O2 uptake was partially or completely suppressed, but itssuppression was removed by DCMU, which indicates that oxygenwas evolved (150 µmol mg–1 Chl hr–1) photosynthetically.We concluded that both photosystems I and II function in guardcell chloroplasts and that these protoplasts have high respiratoryactivity. (Received January 30, 1982; Accepted May 15, 1982)  相似文献   

7.
Inhibition of Photosystem II in Isolated Chloroplasts by Lead   总被引:4,自引:3,他引:1       下载免费PDF全文
Inhibition of photosynthetic electron transport in isolated chloroplasts by lead salts has been demonstrated. Photosystem I activity, as measured by electron transfer from dichlorophenol indophenol to methylviologen, was not reduced by such treatment. However, photosystem II was inhibited by lead salts when electron flow was measured from water to methylviologen and Hill reaction or by chlorophyll fluorescence. Fluorescence induction curves indicated the primary site of inhibition was on the oxidizing side of photosystem II. That this site was between the primary electron donor of photosystem II and the site of water oxidation could be demonstrated by hydroxylamine restoration of normal fluorescence following lead inhibition.  相似文献   

8.
The effects of GA fixation on electron transfers in photosystemsI and II in photosynthesis and energy dependent reactions ofchloroplasts, such as changes in light scattering, H+ uptakeand 515-nm absorbance, were investigated. Fixation of chloroplastswith GA resulted in a lowering of the DCIP and MV photoreductions.DCIP photoreduction activity in fixed chloroplasts was not restoredin the presence of DPC, an electron donor to photosystem II,but was significantly stimulated by DPC when chloroplasts werefixed after aging. The results suggest that the inhibitory effectof GA fixation on photosystem II differs in its mechanism fromthose of treatments such as heating, Tris-washing and aging.The oxidation-reduction reaction of P700 was depressed by GAfixation. Energy dependent reactions in fixed chloroplasts were more markedlydepressed than were electron transfers. Fixed chloroplasts showeda slight conformational response in the presence of PMS. Analysis of the emission spectrum and the induction of chlorophylla fluorescence in fixed chloroplasts suggested that the twopigment systems were partially disordered and that the correspondingprimary photochemical processes were inhibited. (Received November 21, 1972; )  相似文献   

9.
Laminar pulvini of bean (Phaseolus vulgaris L.) contain numerouschloroplasts in cells of their motor tissue. The quantitativerelationships of the chloroplast pigments, chlorophyll a andb, ß-carotene, lutein, neoxanthin as well as the xanthophyllcycle carotenoids (violaxanthin, antheraxanthin and zeaxanthin)were similar to those of mesophyll chloroplasts from leafletlaminae. Exposure of pulvinules to light caused deepoxidationof violaxanthin to zeaxanthin, showing that the xanthophyllcycle is functioning. Chlorophyll fluorescence analysis of pulvinulesconfirmed that their chloroplasts are capable of both photosyntheticelectron transport and non-photochemical fluorescence quenching,showing that they build up a considerable transthylakoid protongradient in the light. Application of DCMU to excised pulvinulesand laminar discs, as well as to pulvinules of intact, attachedterminal leaflets blocked electron transport and fluorescencequenching. Application of the uncoupler CCCP to intact pulvinulesalso prevented non-photochemical fluorescence quenching. Therate of movement of the low-light-adapted terminal leaflet inresponse to exposure of its pulvinule to overhead red light(500 µmol m–2 s–1) was reduced when the pulvinulewas pretreated with DCMU. The pulvinar response to overheadblue light (50 µmol –2 s–1), which is morepronounced than to red light, was not affected by similar pretreatmentwith DCMU. Pretreatment with CCCP caused a short lag in theresponse to red light, but did not affect its subsequent rate.The results suggest that the pulvinar response to red, but notto blue light, requires non-cyclic electron transport and theresulting generation of ATP Key words: Leaf movements, light, non-cyclic electron transport, Phaseolus, pulvinar chloroplasts  相似文献   

10.
Photosynthetic characteristics of detached green barley leavesafter 72 h of treatment with 0·2 mol m–3 of thepyridazinone herbicides SAN 6706, SAN 9785 and SAN 9789 werestudied. For comparison, the effects of 0·01 mol m–3diuron were also investigated. Pyridazinone herbicides causedonly a slight reduction of the total carotene content of thebarley leaves. The total chlorophyll content, as well as thelinolenic to linoleic acid ratio of chloroplast glycerolipids,however, remained unchanged. Diuron treatment caused total inhibitionof electron transport, as revealed by fast fluorescence inductionof leaves and the Hill reaction activity of chloroplasts. The14CO2-nxation by the leaves and the light-induced fluorescencequenching were also completely inhibited in vivo by diuron.Pyridazinone herbicides left 20–40% of the 14CO2-fixationfound in the control, in spite of the fact that their fast fluorescenceinduction tracings showed inhibition in the electron transport.Chloroplasts isolated from the leaves treated with pyridazinoneswere found to be highly active in mediating the ferricyanide-dependentHill reaction. In order to test the ability of pyridazinonesto inhibit photosynthetic electron transport in vivo, their‘prompt’ effect on fluorescence was also investigated.It is concluded that pyridazinone herbicides can readily andrapidly enter the chloroplasts and inhibit the photosyntheticelectron transport in vivo. The differences between the long-termeffects of pyridazinones and those of diuron suggest differencesin the inhibitory effectiveness on the various photosyntheticparameters between the two herbicide groups. It is suggestedthat pyridazinones can leave the chloroplasts during isolationowing to the loose binding onto the thylakoid membranes. Key words: Pyridazinone herbicides, electron transport, fluorescence induction  相似文献   

11.
Critchley C 《Plant physiology》1981,67(6):1161-1165
Cucumber plants (Cucumis sativus L.), grown at low quantum flux density (120-150 microeinsteins per square meter per second) were photoinhibited by a three-hour exposure in air to ten times the light intensity experienced during growth. Chloroplasts were isolated from photoinhibited and control leaves and the following activities determined: O2 evolution in the presence of ferricyanide, photosystem I activity, noncyclic and cyclic photophosphorylation, and light-induced proton uptake. Chlorophyll and chloroplast absorbance spectra, and chloroplast fluorescence were also measured. It was found that photosystem II electron transport and non-cyclic photophosphorylation were inhibited by about 50%, while cyclic photophosphorylation was less inhibited and photosystem I electron transport and light-induced proton uptake were unaffected. Electron transport to methylviologen could not be fully restored by electron donation to photosystem II. Chloroplast fluorescence induction at room temperature was strongly reduced following photoinhibition. There was no difference in the absorption spectra of the extracted chlorophylls from control and photoinhibited chloroplasts, but an increase of the absorption in the blue wavelength region was observed in the photoinhibited chloroplasts. It is suggested that high light stress does not result in alteration of the membrane properties, as is the case in low-temperature stress for example, but affects directly the photosynthetic reaction centers, primarily of photosystem II.  相似文献   

12.
Normal Euglena chloroplasts contained 1 atom of Mn per 47±8chlorophyll molecules. The manganese content of chloroplastswas decreased by heat treatment. After complete removal of manganeseby incubation at 45°C for 5 min, Hill activity with DPIPas electron acceptor was abolished, but the activity of DPIPphotoreduction with diphenylcarbazide as electron donor wasunaffected. Hill activity was inactivated by incubating Euglena chloroplastsat alkaline pH. The presence of a high concentration of Trisduring incubation of chloroplasts at an alkaline pH had no additionaleffect on the activity drop. Donor-supported DPIP photoreduction in heated Euglena chloroplasts,as well as the normal Hill reaction in untreated chloroplasts,was inhibited by DCMU, HOQNO and ioxynil which block electrontransport at the reducing side of system II. These reactionswere also inhibited by another group of inhibitors; CCCP, salicylaldoxime,antimycin A and azide, which block electron transport at a sitebetween the electron carriers, Y1 and Y2 located on the oxidizingside of system II. Possible sites of inhibition by heat treatment and by inhibitorsand sites for entry of electrons from artificial electron donorsin the photosynthetic electron transport chain, especially inrelation to the functional site of endogenous manganese in chloroplasts,were proposed. (Received October 30, 1971; )  相似文献   

13.
The kinetics of 685 nm chlorophyll fluorescence emission weremeasured at 20 °C following illumination of primary leavesof P. vulgaris. During foliar senescence, a large reductionwas observed in the maximal level of fluorescence emission (P)of the induction curve, normalized with respect to the minimallevel (O), and in the time taken to reach P. This suggests thatfewer plastoquinone (PQ) molecules were able to accept electronsfrom each photosystem two (PS II) reaction centre in older leaves.Measurements of fluorescence emission at 77 °K indicatedthat the primary photochemical quantum yield of the PS II reactioncentres remained constant during senescence. The redox stateof the PQ pool was estimated throughout the induction curveat 20 °C. In both mature and senescent leaves PQ was highlyreduced at P. There followed a reoxidation of PQ in the matureleaves, but in the old leaves the PQ pool remained reduced.This indicates that the rate of electron flow from PQ to photosystemone (PS I) decreased considerably during senescence. Fluorescencewas quenched from P to a steady state level (T) in leaves ofall ages, and this was associated with a redistribution of excitationin favour of PS I. Since, in senescent leaves, changes in theredox state of PQ were absent, it is suggested that quenchingresulted from the generation of proton and ion gradients acrossthe thylakoid membranes, and the synthesis of ATP.  相似文献   

14.
Chemiluminescence of luminol (CLL) was induced by illuminatedspinach chloroplast fragments. CLL was diminished by superoxidedismutase or under anaerobic conditions and increased by anautoxidizable electron acceptor, methyl viologen. The optimumpH for CLL was 10.0-10.5. Ferredoxin and cytochrome c reducing substance (CRS) did notaffect the intensity of CLL, but accelerated the dark decayin the absence of methyl viologen. In the presence of methylviologen, ferredoxin and CRS lowered the intensity and acceleratedthe dark decay. 3-(4-Chlorophenyl)-1,1-dimethylurea diminishedCLL. Carbonylcyanide m-chlorophenylhydrazone accelerated theinitial rate of CLL increase at low concentration and inhibitedit at high concentration. Half-decay time of CLL after the cessationof light was shortened by inhibiting electron transfer on theoxidizing side of photosystem II. We conclude that most of the CLL observed in illuminated chloroplastsis dependent on O2. The results also suggest that O2is reduced by reduced ferredoxin or CRS and oxidized on theoxidizing side of photosystem II. The half life of O2in illuminated chloroplasts was estimated from the half-decaytime of CLL to be a few sec. 1 Present address: Kyushu Dental College, Department of Biology,Kitakyushu 803, Japan. (Received May 30, 1977; )  相似文献   

15.
The kinetics of changes in photosystem I (PSI), photosystemII (PSII), and whole chain (PSII and PSI) electron transport,chlorophyll fluorescence parameters, the capacity to bind atrazineand the polypeptide profiles of thylakoids isolated from wheatleaves on exposure to a photon flux density of 2000 µmolm–2 s–1 were determined. Severe and similar levelsof photo-inhibitory damage to both PSII and whole chain electrontransport occurred and were correlated with decreases in theratio of variable to maximal fluorescence, the proportionalcontribution of the rapid a phase of the fluorescence kineticsand the capacity to bind atrazine. Severe photo-inhibition ofelectron transport was not associated with a major loss of chlorophyllor total thylakoid protein. However, a small decrease in a 70kDa polypeptide together with increases in a number of low molecularmass polypeptides (8–24 kDa) occurred. Phosphorylation of thylakoid polypeptides alleviated photo-inhibitionof PSII electron transport but stimulated photoinhibitory damageto whole chain electron transport. The consequences of suchphosphorylation-induced effects on photoinhibition in vivo areconsidered. Key words: Chlorophyll fluorescence, electron transport, photo-inhibition, protein phosphorylation, thylakoid membranes, wheat (Triticum aestivum)  相似文献   

16.
Chloroplasts from 17-d-old pea leaves (Pisum sativum L.) wereisolated to elucidate the requirements for the light-induceddegradation of stromal proteins. The influence of electron transportthrough the thylakoids and the influence of ATP on protein degradationwere investigated. When chloroplasts were incubated in the light(45 µmol m–2s–1), glutamine synthetase, thelarge subunit of ribulose-1,5-bisphosphate carboxylase and glutamatesynthase were degraded, whereas phosphoribulokinase, ferredoxin-NADP+reductase and the 33 kDa protein of photosystem II remainedmore stable. Major protein degradation was not observed over240 mm in darkness. The electron transport inhibitor dichlorophenyldimethylureareduced protein degradation in the light over several hours,whereas dibromothymoquinone was less effective. Inhibiting theproduction of ATP with tentoxin or by destroying the  相似文献   

17.
We have investigated submillisecond delayed luminescence in spinach chloroplasts under a variety of conditions. In Tris-washed chloroplasts, which are inhibited on the oxidizing side of P-680, the delayed light emission in the 7–200 μs time-range decayed with biphasic behavior. In fully dark-adapted samples illuminated by a single saturating laser pulse, the fast phase of delayed luminescence followed a nearly identical pH-dependent time-course as that observed optically and by ESR for P+-680 reduction, thus verifying the recombination hypothesis for the origin of delayed light. The observed slower phase of delayed luminescence was also pH dependent, but unlike the fast phase, could not be ascribed to specific electron transfer events of PS II. This phase could be rationalized by a heterogeneity in the population of P-680. While kinetic parameters were found to be insensitive to changes in ionic strength, the overall luminescence intensity was quite sensitive to the electrical parameters, thus indicating the role of ionic strength and local charges in delayed luminescence modulation. A similar series of experiments was performed on untreated chloroplasts. The pH-dependent delayed luminescence behavior in both untreated chloroplasts and Tris-washed chloroplasts was similar despite significantly faster kinetics associated with the reduction of P+-680 by the secondary PS II electron donor, Z, in the former preparation (e.g., Van Best, J.A. and Mathis, P. (1978) Biochim. Biophys. Acta 503, 178–188). Thus, it was concluded that, in untreated samples, microsecond delayed luminescence emanates primarily from centers which are not competent in oxygen evolution. The nearly identical delayed luminescence intensity in untreated chloroplasts and in Tris-washed chloroplasts was rationalized by a model which predicts modulations in delayed luminescence yield by the exciton-quenching effect of P+-680. Computer simulations demonstrate the feasibility of this model. The previously documented flash oscillations in microsecond delayed luminescence intensity in untreated chloroplasts (Bowes, J.M. and Crofts, A.R. (1979) Biochim. Biophys. Acta 547, 336–346), which we readily observed, were attributed to alterations in delayed luminescence yield (in nonfunctional centers) by variations in charge density stored at the oxygen-evolving complex of functional centers. Taken together, our results emphasize the dependence of delayed luminescence kinetics upon electron-transfer kinetics and the dependence of delayed luminescence amplitude upon the photochemical parameters, the exciton yield and the emission yield.  相似文献   

18.
The effect of heat stress on photosynthetic electron transportwas investigated in thylakoids isolated from the wheat (Triticumaestivum L.) varieties APU (Finland) and K65 (India) grown underboth cool (13 °C day, 10 °C night) and warm (30 °Cday, 25 °C night) regimes which gave rise to varietal differencesin photosynthetic temperature acclimation. The responses ofthe uncoupled activities of both whole-chain electron transportand photosystem II to heat stress were similar. Both activitiesexhibited higher rates in thylakoids isolated from warm-grownplants and were more resistant to high temperature pretreatmentthan in those isolated from cool-grown plants, but varietaldifferences were not observed. Uncoupled photosystem I activity driven by either reduced 2,6-dichlorophenol indophenol (DCPIPH2) or N,N,N',N'-tetramethyl-p-phenylenediamine (TMPDH2) showed a stimulation following high temperaturepretreatment which was more marked in thylakoids isolated fromwarm-grown plants, followed by inhibition at extreme high temperatures.This stimulation was due largely to an increase in Vmax butdid not occur when reduced diaminodurene, which is highly lipophilic,was used as the electron donor. It appears that stimulationof PS I activity may involve increased accessibility of someartificial electron donors to the native acceptor sites withinthe thylakoid membrane in a process which is influenced by growthtemperature. Key words: Photosynthetic electron transport, heat stress, Triticum aestivum  相似文献   

19.
Hardt H  Kok B 《Plant physiology》1977,60(2):225-229
Treatment of spinach chloroplasts with glutaraldehyde causes an inhibition in the electron transport chain between the two photosystems. Measurements of O2 flash yields, pH exchange, and fluorescence induction show that the O2 evolving apparatus, photosystem II and its electron acceptor pool are not affected. The behavior of P700 indicates that its reduction but not its oxidation, is severely inhibited. Cytochrome f is still reducible by photosystem II but also slowly oxidizable by photosystem I. The sensitivity of isolated plastocyanin to glutaraldehyde further supports the conclusion that glutaraldehyde inhibits at the plastocyanin level and thereby induces a break between P700 and cytochrome f.  相似文献   

20.
Fluorescence transients were investigated with the diatom Phaeodactylumtricornutum. Supplementary experiments were done with Chaetocerossp. Under weak excitation ({small tilde}103 erg/cm2sec), fluorescencetransients were induced simply by die oxidation-reduction reactionof Q, the primary reductant of photosystem II. The action spectraindicated that the electron transfer components between thetwo photosystems were in the most reduced state when fucoxanthinwas excited. The transients were observed with the 681 run emissionand with the 707 nm emission at room temperature. At –196°C,induction due to the reduction of Q. appeared both at the 681and 707 nm emissions. Similar results were also obtained withChaetoceros sp. Under strong excitation (104–105 erg/cm2-sec), the fluorescencetransients due to the interconversion between States 1 and 2of die pigment system (cf. ref. 27, 29) were observed. The transientswere induced by die alternate excitation of chlorophyll a andfucoxanthin or chlorophyll c. Conversion from State 2 to State1 was inhibited by DNP and CCCP, indicating that die processwas energy-dependent. Fluorescence spectra at –196°Cwere not altered by die state-conversion of die pigment system. These results suggest diat all die fluorescence bands whichappeared at room temperature and at –196°C were dueto die chlorophyll a of pigment system II in Phaeodactylum andChaetoceros. (Received September 7, 1972; )  相似文献   

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