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1.
The redox kinetics of P700 induced by far-red light and a pulseof strong white light in wild type cells were compared withthose in NAD(P)H dehydrogenase (NDH)-defective mutants of thecyanobacterium Synechocystis PCC 6803. The wild type cells showedthe electron donation from the respiratory donor and the photoreductantgenerated in PS I to P700+ through the plastoquinone, whichis mediated by a Hg2+-sensitive enzyme. The NDH-defective mutantcells, however, did not show the electron transfer to P700+through the plastoquinone from both the photoreductant in PSI and cytosolic electron donors using pyndine nucleotides asan intermediate. Thus, NDH appears to be the site of main entryof electrons into the plastoquinone pool in the NAD(P)H-mediatedcyclic electron flow and the respiratory electron flow in Synechocystis. (Received August 31, 1992; Accepted October 1, 1992)  相似文献   

2.
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)  相似文献   

3.
A method for estimation of the functional pool size of electronsthat can be donated to P700+ after the illumination of intactleaves with actinic light is described. The complementary areasbetween the stationary level of P700+ attained by irradiationwith far-red light and the oxidation curves of P700 by far-redlight after a 50-ms multiple-turnover light (MT-area), and afterthe illumination of actinic light (AL-area) were determined.Since the MT-area represents the pool size of electrons in theintersystem chain, the ratio of the AL-area to the MT-area allowsus to estimate the pooi size of electrons stored during actinicillumination that can be donated to P700+ The ratio of the AL-areato the MT-area was determined for intact leaves of several C3plants, and it was found to increase to a value of two or threewith increases in the intensity of actinic light, and it wasfurther increased by anaerobiosis. The pool size of electronsthat can be donated to P700+ from the stroma was estimated tobe in the range of 12 and 28 electrons per P700 under aerobicconditions. These results indicate that stromal components donateelectrons to P700+ through the intersystem chain in chloroplastsof C3 plants. (Received July 3, 1992; Accepted September 29, 1992)  相似文献   

4.
The functional pool size of electrons in the intersystem chainof the chloroplasts of maize was estimated to be about 25 perP700 by the redox change in P700 with single- and multiple-turnoverlights under far-red light in intact leaves. This is about twicethe pool size observed in C3 plants. Furthermore, the stromalpool size of electrons that can be donated to P700+ after actinicillumination was larger in maize leaves than in leaves of C3plants, giving a maximum value of 225 electrons per P700. Maizeleaves showed an increase in the yield of modulated Chl fluorescenceafter turning off of actinic light, which confirms the donationof electrons in the dark to the intersystem chain from the stromaldonors that accumulated during actinic illumination. We proposethat the mesophyll chloroplasts are responsible for a high levelof electron-donating activity to the intersystem chain fromstromal donors such as triose phosphates and malate with NADPHas an intermediate. The level of P700+ under strong far-redlight was decreased after actinic illumination, suggesting theoperation of an actinic light-triggered cyclic electron flowin chloroplasts of the bundle sheath cells. (Received August 14, 1992; Accepted October 13, 1992)  相似文献   

5.
Stability of thylakoid components under supra-high irradiancewas studied with the cyanophyte Synechocystis PCC 6714. Theactivity of overall photosynthesis was quickly inactivated (T1/2=20min) under supra-high irradiance (300 W m–2, white light).In parallel with the inactivation of photosynthesis, QA in PSII was also inactivated. Both inactivations were acceleratedby chloramphenicol (CAP) addition. The reactivation of PS IIrequired weak irradiation and was suppressed by CAP. However,PS I measured as P700 was very stable. The level of PS I measuredas P700 was not significantly reduced by the irradiation for12 h even in the presence of CAP while the level of Cyt b559,component of PS II, was decreased markedly. The function ofPS I before and after supra-high irradiation with CAP was examinedby comparing sizes of P700 oxidation induced by a short flash,by a continuous light, and by determination of O2-and ferredoxin-reduction.No difference was observed in PS I actions before and afterthe irradiation treatment. These results indicate that the PSI complex is very tolerant of supra-high irradiation. However,the cells grown under supra-high irradiance contained much fewerPS I and PS II complexes than Cyt b6–f complexes. Theformer levels were reduced to a half to one fourth of thosebefore growth while the level of Cyt b6–f complex wasnot reduced so much. A possible mechanism for changes in thylakoidcomposition under supra-high irradiation was discussed. (Received February 16, 1991; Accepted June 12, 1991)  相似文献   

6.
By treating a FA/FB-depleted P700-Fx core from SynechococcusPCC 6301 with diethylether, most of the phylloquinone was removedwithout loss of P700. The 1 ms decay of P700+ in the originalcore was replaced by the 25 ns decay, which was interpretedas the backreaction occurring in a P700+  相似文献   

7.
Changes in the activity of cytochrome c oxidase (EC 1.9.3.1 [EC] ,Cyt-oxidase) in response to growth conditions were studied withthe cyanophyte Synechocystis PCC 6714 in relation to changesin PSI abundance induced by light regime for photosynthesis.The activity was determined with the Vmax of mammalian cytochromec oxidation by isolated membranes. The activity of glucose-6-phosphate(G-6-P):NADP+ oxidoreductase (EC 1.1.1.49 [EC] ) was also determinedsupplementarily. Cyt-oxidase activity was enhanced by glucoseadded to the medium even when cell growth maintained mainlyby oxygenic photosynthesis. G-6-P:NADP+ oxidoreductase was alsoactivated by glucose. The enhanced level of Cyt-oxidase washigher under PSII light, which causes high PSI abundance, thanthat under PSI light, which causes low PSI abundance. The levelwas intermediate under hetetrotrophic conditions. Although theactivity level was low in cells grown under autotrophic conditions,the level was again lower in cells grown under PSI light thanunder PSII light. The change of Cyt-oxidase activity in responseto light regime occurred in the same direction as that for thevariation of PSI abundance. Results suggest that in SynechocystisPCC 6714, the capacity of electron turnover at the two terminalcomponents of thylakoid electron transport system, Cyt-oxidaseand PSI, changes in parallel with each other in response tothe state of thylakoid electron transport system. 1Present address: Institute of Botany, Academia Sinica, Beijing100044, China 2Present address: Department of Botany, Utkal University, Bhubaneswar,India 751004  相似文献   

8.
The effect of Na+ on phosphate uptake was studied in four strainsof cyanobacteria: Synechococcus PCC 7942, Gloeothece PCC 6501,Phormidium sp. and Chlorogloeopsis PCC 6912. Phosphate uptakewas stimulated by Na+ in all cases. Li+ and K+ acted as partialanalogues for Na+. Half-saturation [K1/2(Na+)] of phosphateuptake was reached with Na+ concentrations ranging from 317µM in Chlorogloeopsis to 659 µM in Phormidium. Theconcentration of phosphate required to reach half-saturationof phosphate uptake [K1/2(Pi)]was not changed by the presenceof Na+. (Received April 11, 1994; Accepted July 5, 1994)  相似文献   

9.
Internode elongation was measured in plants of Phaseolus vulgarisand Glycine max grown under 8 h photoperiods at 25 W m–2in white fluorescent light, followed by light-extensions varyingin quality, irradiance and duration. Two distinct responsesto light were observed under these conditions. A reduction in PFR/P increased elongation, but elongation wasalso modified by a second reaction in which internode lengthincreased with increase in the duration and irradiance of theday-extension. This light-promoted response occurred in bothred and blue light. In the PFR-inhibition response, light acteddirectly on the expanding internode. The light-promoted response,in contrast, required irradiation of the leaf. The response to a short end-of-day exposure to far-red lightprogressively diminished as successive internodes expanded underthe treatment, whereas the light-promoted response increased.The two processes appeared to interact and, in the later-expandinginternodes, the effect of a reduction in PFR was greater underlong day-extensions with mixed red and far-red light than inthe end-of-day treatments. 1 Present address: British Telecom, Brunel House, 2 FitzalanRoad, Cardiff, U.K.  相似文献   

10.
Bundle sheath chloroplasts of NADP-malic enzyme (NADP-ME) type C4 species have a high demand for ATP, while being deficient in linear electron flow and oxidation of water by photosystem II (PSII). To evaluate electron donors to photosystem I (PSI) and possible pathways of cyclic electron flow (CEF1) in isolated bundle sheath strands of maize (Zea mays L.), an NADP-ME species, light-induced redox kinetics of the reaction center chlorophyll of PSI (P700) were followed under aerobic conditions. Donors of electrons to CEF1 are needed to compensate for electrons lost from the cycle. When stromal electron donors to CEF1 are generated during pre-illumination with actinic light (AL), they retard the subsequent rate of oxidation of P700 by far-red light. Ascorbate was more effective than malate in generating stromal electron donors by AL. The generation of stromal donors by ascorbate was inhibited by DCMU, showing ascorbate donates electrons to the oxidizing side of PSII. The inhibitors of NADPH dehydrogenase (NDH), amytal and rotenone, accelerated the oxidation rate of P700 by far-red light after AL, indicating donation of electrons to the intersystem from stromal donors via NDH. These inhibitors, however, did not affect the steady-state level of P700+ under AL, which represents a balance of input and output of electrons in P700. In contrast, antimycin A, the inhibitor of the ferredoxin-plastoquinone reductase-dependent CEF1, substantially lowered the level of P700+ under AL. Thus, the primary pathway of ATP generation by CEF1 may be through ferredoxin-plastoquinone, while function of CEF1 via NDH may be restricted by low levels of ferredoxin-NADP reductase. NDH may contribute to redox poising of CEF1, or function to generate ATP in linear electron flow to O2 via PSI, utilizing NADPH generated from malate by chloroplastic NADP-ME.  相似文献   

11.
P700 was enriched relative to antenna pigments by treating thethylakoid membranes from thermophilic cyanobacterium (Synechococcussp.) with diethylether. The total Chi a/P700 ratio of the membraneswas 147 but decreased to 13 after the treatment with ether whichhad been 70% saturated with water. Vitamin K1 and carotenoidswere completely removed by this treatment. The low chlorophylla to P700 ratio was retained in photosystem I reaction-centercomplexes purified from the ether-extracted membranes with TritonX-100. The midpoint potential of P700 was considerably loweredby the ether treatment of the thylakoid membranes but was partiallyreversed by the further treatment of the extracted membraneswith Triton X-100. Photo-oxidation of P700 in the purified complexeswas extremely slow under continuous illumination, indicatingthat there is no significant leakage of electrons from the primaryelectron acceptor to other bound acceptors or O2. The photooxidationof P700 was appreciably accerelated on addition of vitamin K3(or K1). (Received June 23, 1988; Accepted November 25, 1988)  相似文献   

12.
The origin of nonmonotonic changes in the redox state of P700, the primary electron donor of PSI, was investigated on predarkened barley (Hordeum vulgare L.) leaves exposed to far-red light. To accomplish this, the relaxation kinetics of absorbance changes at 830 nm, reflecting the dark reduction of P700+, were measured at different stages of the induction curve. The onset of far-red light resulted in rapid oxidation of P700, which was followed by its partial reduction and subsequent slow oxidation of P700 to a steady-state level. This steady-state level was usually attained within 10 s under far-red light. The relative contribution of the slow kinetic component of P700+ reduction decreased in parallel with the transient photoreduction of P700+ and increased upon a subsequent stage of P700 photooxidation. The contribution of the middle component to the dark reduction of P700+ increased monotonically with the length of far-red light irradiation. The relative amplitude of the fast component of P700+ reduction increased sharply during the first 3 s of irradiation and decreased upon longer light exposures. The rates of fast and slow components of dark reduction of P700+ remained constant upon illumination of dark-adapted leaves with far-red light for 1 s and longer periods. Thus, nonmonotonic changes in the redox state of P700 in barley leaves exposed to far-red light reflect variable contributions of few alternative electron transport pathways characterized by different rates of electron donation to PSI. The results show the principle possibility of switching-over between alternative pathways of PSI-related electron transfer within one complex of this photosystem. Such switching may occur irrespective of active operation or inhibition of ferredoxin-dependent electron transport.  相似文献   

13.
Changes in photosystem stoichiometry in response to shift ofenvironments for cell growth other than light regime were studiedwith the cyanophyte Synechocystis PCC 6714 in relation to thechange induced by light-quality shift. Following two environment-shiftswere examined: the shift of molecular form of inorganic carbonsource for photosynthesis from CO2 to HCO3 (CO2 stress)and the increase in salinity of the medium with NaCl (0.5 M)(Na+ stress). Both CO2 and Na+ stresses induced the increasein PSI abundance resulting in a higher PSI/PSII stoichiometry.CO2 stress was found to elevate simultaneously Cyt c oxidaseactivity (Vmax). The feature was the same as that caused bylight-quality shift from preferential excitation of PSI to PSII(light stress) though the enhancement by either stress was smallerthan that by light stress. Under our experimental conditions,PSI/PSII stoichiometry appeared to increase at a fairly constantrate to the basal level even when the basal level had been differentlydetermined by the light-quality. Enhancing rates for PSI/PSIIstoichiometry and for Cyt c oxidase activity were also similarto each other. Since the two stresses affect the thylakoid electrontransport similarly to the shift of light-quality, we interpretedour results as follows: three environmental stresses, CO2, Na+,and light stresses, cause changes in electron turnover capacityof PSI and Cyt c oxidase under a similar, probably a common,mechanism for monitoring redox state of thylakoid electron transportsystem. 1On leave from Department of Biology, College of Natural Science,Kyngpook National University, Taegu 702-701, Korea. 2Present address: Department of Marine Bioscience, Fukui Pre-fecturalUniversity, Obama, Fukui, 917 Japan.  相似文献   

14.
Transient absorbance changes of the primary electron donor chlorophylla (P680) and acceptor pheophytin a (H) were measured at 77 Kby nanosecond laser spectroscopy in the D1-D2-cytochrome b559photosystem II reaction center complex containing dibromomethylisopropylbenzoquinone (DBMIB). After the laser excitation of the reactioncenter in the presence of DBMIB, only the P680+-(DBMIB-) statewas detected. P680+ mainly decayed with a t1/e of 11 ms. Inthe absence of DBMIB, the excitation produced the P680+H- radicalpair. The radical pair produced the triplet state (P680T) witha t1/e of 50 ns, and P680T then decayed with a t1/e of 2.1 ms.It was concluded that H- was oxidized by DBMIB in a time rangefaster than the detecting time resolution (3.5 ns) even at 77K. The rapid oxidation of H- by DBMIB was also confirmed bythe suppression of delayed fluorescence with a decay t1/e of50 ns. The P680+(DBMIB-)/P680(DBMIB) difference spectrum exhibiteda Qy, band with a peak at 682 nm with a shoulder at 673 nm.The spectral shape was almost temperature insensitive between77 and 265 K. The feature of this spectrum in the wavelengthrange between 330 and 720 nm was compared with that of P680T/P680or H-/H at 77 K. (Received May 8, 1996; Accepted June 24, 1996)  相似文献   

15.
Light-induced redox-reactions of cytochrome b559 in spinachchloroplasts were investigated. Illumination of chloroplastsinduced photoreduction of cytochrorne b559 Red light (650 nm)was more effective than far-red light (725 nm), indicating thatthe photoreduction is a photosystem II-mediated reaction. Onaddition of DCMU, the photoreduction was eliminated and a photooxidationof cytochrome b559 was observed. The rate of this photooxidationwas faster with photosystem II light than with photo-systemI light. On addition of Mn++ the photooxidation was partly suppressed;far-red light became as effective as red light in inducing photooxidationof cytochrome b599, in the presence of DCMU and Mn++. Ascorbate completely suppressed photooxidation of cytochromeb559 In the presence of ascorbate, however, photooxidation wasobserved in the presence of inhibitors or after inhibitory treatmentsof chloroplasts which affected the oxidizing side of systemII. These inhibitors and inhibitory treatments, but not DCMU,decreased the redoxpotential of cytochrome b559. Reactivationof Hill reaction in Tris-washed chloroplasts by indophenol-ascorbatetreatment was not accompanied by an abolishment of photooxidationof cytochrome b559. A possible mechanism is proposed to account for these reactionsof cytochrome b559 in the photosynthetic electron transportin chloroplasts. (Received April 4, 1972; )  相似文献   

16.
We have reported that ryanodine receptor (RyR) channels display three different responses to cytoplasmic free Ca2+ concentration ([Ca2+]) depending on their redox state (Marengo JJ, Hidalgo C, and Bull R. Biophys J 74: 1263–1277, 1998), with low, moderate, and high maximal fractional open times (Po). Activation by ATP of single RyR channels from rat brain cortex was tested in planar lipid bilayers with 10 or 0.1 µM cytoplasmic [Ca2+]. At 10 µM [Ca2+], low-Po channels presented lower apparent affinity to activation by ATP [[ATP] for half-maximal activation (KaATP) = 422 µM] than moderate-Po channels (KaATP = 82 µM). Oxidation of low-Po channels with thimerosal or 2,2'-dithiodipyridine (DTDP) gave rise to moderate-Po channels and decreased KaATP from 422 to 82 µM. At 0.1 µM cytoplasmic [Ca2+], ATP induced an almost negligible activation of low-Po channels. After oxidation to high-Po behavior, activation by ATP was markedly increased. Noise analysis of single-channel fluctuations of low-Po channels at 10 µM [Ca2+] plus ATP revealed the presence of subconductance states, suggesting a conduction mechanism that involves four independent subchannels. On oxidation the subchannels opened and closed in a concerted mode. subconductance states; calcium ion release channels; calcium ion regulation; thimerosal; 2,2'-dithiodipyridine  相似文献   

17.
At constant external [CO2], rates of dark-CO2 fixation of theunicellular green alga Eremosphaera viridis were drasticallyincreased (up to 40-fold) by addition of ammonium (NH3+ NH4+)at external pH values (pH0) between 6.0 and 8.0. The cytosolicpH was monitored under identical conditions by micro-pH-electrodemeasurements, and cytosolic and vacuolar pH by the 31P-NMR technique.Addition of ammonium (5.0 mol m pH0 7.0) caused a rapidand transient acidification of the cytosol during the first4 min. Thereafter, the cytosolic pH remained constant at itsoriginal value. A rather constant cytosolic pH was also confirmedby 31P-NMR measurements, which, in addition, indicated a slowalkalization of the vacuole (about 0.5 units within 30 min afteraddition of ammonium). Since the dramatic stimulation of dark-CO2 fixation by ammoniumis not mediated by an alkalization of the cytosol, nor by directammonium effects on phosphoenolpyruvate carboxylase (PEPC, E.C.4.1.1.31 [EC] ), the role of vacuolar alkalization as a possible triggerfor the stimulation of PEP-carboxylase is discussed. Key words: Cytosolic pH, dark-CO2 fixation, pH-regulation, vacuolar pH  相似文献   

18.
The methyl viologen (MV)-dependent, linear electron flow fromPS II to PS I was severely blocked in intact or broken, uncoupledchloroplasts when oxygen was removed from the suspension medium,as revealed by measurements of chlorophyll fluorescence andthe rate of photoreduction of MV. Kinetics of the reductionof pre-oxidized P700 by a saturating light pulse showed thatreduced MV in the absence of oxygen re-reduces P700+ via theintersystem electron transport chain. Since the re-reductionof P700+ was inhibited by 2,5-dibromo-3-methyl-6-isopropyl-p-benzoquinone,the MV-mediated cyclic electron flow, in contrast to the phenazinemethosulphate-catalyzed one, involves the plastoquinone pool.However, 2-n-heptyl-4-hydro-xyquinoline-N-oxide, 2-n-nonyl-4-hydroxyquinoline-N-oxideand antimycin A did not inhibit the MV-mediated flow. Thus,the inhibition of the linear electron flow in chloroplasts underanaerobic conditions suggested the overreduction of the plastoquinonepool as a result of the MV-mediated cyclic flow (Received February 13, 1990; Accepted March 31, 1990)  相似文献   

19.
Etiolated Hordeum vulgare (barley) Plants were greened underwhite light or far-red (> 700 nm) light. Exposure to far-redlight inhibited chlorophyll synthesis (especially chlorophyllb) and the development of photosystem II which were seen whengreening took place under white light. Primary leaves were detachedand the labelling of their acyl lipids from [14C]acetate wasstudied under white light or far-red light illumination. Greeningwith far-red light caused a reduction in the radiolabellingof polyunsaturated fatty acids and diacylgalactosylglycerol.Total fatty acid labelling rates were unaffected. Phosphatidylethanolamine,which was normally poorly labelled, accounted for up to 15 percent of the total radioactivity in acyl moieties of lipids inleaves greened with far-red light. The results are discussedin connection with the role that acyl lipids may play in normalthylakoid structure and function. Hordeum vulgare, barley, acyl lipids, white light, far-red light, chloroplasts, thylakoids  相似文献   

20.
In a colorless mutant of Chlorella kessleri, far-red light significantlyenhanced the K+ efflux. This effect was abolished by the K+channel-blocker tetraethylammonium acetate. Using cyanine dyeto monitor membrane potential, we deduced that the K+ effluxunder far-red light was probably accompanied by hyperpolarizationof the plasmalemma. (Received August 30, 1993; Accepted November 16, 1993)  相似文献   

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