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2.
A model for the photochemical apparatus of photosynthesis is presented which accounts for the fluorescence properties of Photosystem II and Photosystem I as well as energy transfer between the two photosystems. The model was tested by measuring at ?196 °C fluorescence induction curves at 690 and 730 nm in the absence and presence of 5 mM MgCl 2 which presumably changes the distribution of excitation energy between the two photosystems. The equations describing the fluorescence properties involve terms for the distribution of absorbed quanta, α, being the fraction distributed to Photosystem I, and β, the fraction to Photosystem II, and a term for the rate constant for energy transfer from Photosystem II to Photosystem I, kT(II→I). The data, analyzed within the context of the model, permit a direct comparison of α and kT(II→I) in the absence (?) and presence (+) of Mg 2+:α/ ?α += 1.2and k/ ?T(II→I)k +T(II→I)= 1.9. If the criterion thatα + β = 1 is applied absolute values can be calculated: in the presence of Mg 2+, a+ = 0.27 and the yield of energy transfer,φ +T(II→I) varied from 0.065 when the Photosystem II reaction centers were all open to 0.23 when they were closed. In the absence of Mg 2+,α ? = 0.32 andφ T(II→I) varied from 0.12 to 0.28.The data were also analyzed assuming that two types of energy transfer could be distinguished; a transfer from the light-harvseting chlorophyll of Photosystem II to Photosystem I, kT(II→I), and a transfer from the reaction centers of Photosystem II to Photosystem I, kt(II→I). In that caseα/ ?α += 1.3, k/ ?T(II→I)k +T(II→I)= 1.3 and k/ ?t(II→I)k +(tII→I)= 3.0. It was concluded, however, that both of these types of energy transfer are different manifestations of a single energy transfer process. 相似文献
3.
The electroluminescence induced by external electric fields in blebs prepared from chloroplasts consists of two kinetically different phases, rapid (R) and slow (S), which were shown to be linked to Photosystem I (PS I) and Photosystem II (PS II) activities, respectively (Symons, M., Korenstein, R. and Malkin, S. (1985) Biochim. Biophys. Acta 806, 305–310). In this report we describe conditions involving heat treatment of broken chloroplasts, which make it possible to observe R phase electroluminescence essentially devoid of any contribution by the S phase. This allowed the precise measurement of the emission spectrum of PS I electroluminescence. The emission spectrum of PS II electroluminescence was obtained using regular broken chloroplasts, which show only S-type emission. The latter emission spectrum is identical to the one obtained for ordinary prompt fluorescence, peaking at 685 nm with a bandwidth of about 25 nm. The PS I emission spectrum is symmetric around 705 nm and is much broader, about 60 nm. 相似文献
4.
The ratio of Photosystem (PS) II to PS I electron-transport capacity in spinach chloroplasts was compared from reaction-center and steady-state rate measurements. The reaction-center electron-transport capacity was based upon both the relative concentrations of the PS II α, PS II β and PS I centers, and the number of chlorophyll molecules associated with each type of center. The reaction-center ratio of total PS II to PS I electron-transport capacity was about 1.8:1. Steady-state electron-transport capacity data were obtained from the rate of light-induced absorbance-change measurements in the presence of ferredoxin-NADP +, potassium ferricyanide and 2,5-dimethylbenzoquinone (DMQ). A new method was developed for determining the partition of reduced DMQ between the thylakoid membrane and the surrounding aqueous phase. The ratio of membrane-bound to aqueous DMQH 2 was experimentally determined to be 1.3:1. When used at low concentrations (200 μM), potassium ferricyanide is shown to be strictly a PS I electron acceptor. At concentrations higher than 200 μM, ferricyanide intercepted electrons from the reducing side of PS II as well. The experimental rates of electron flow through PS II and PS I defined a PS II/PS I electron-transport capacity ratio of 1.6:1. 相似文献
5.
Mutant C-2A' of Scenedesmus obliquus formed only traces of chlorophylland showed no detectable photosynthesis when grown heterotrophically.When transferred to light this mutant developed chlorophylland its photosynthetic capacity was established. Following ashort initial lag phase, both photosynthetic capacity and totallight absorption of the cells reached saturation more rapidlythan did the rate of chlorophyll synthesis. Consequently, thequantum requirement of photosynthesis showed a rapid declineduring the initial 6 hr of greening, to a best value of 8. Subsequently,a slow increase occurred as additional chlorophyll was synthesized.Behavior parallel to that of the quantum requirement was alsonoted for the relative fluorescence yield and for the onsetof the 520 nm light-induced absorbance change. Of the two photosystems,PS-I seemed to develop more rapidly than did PS-II. The appearanceof PS-II activity appeared to accompany linkage of the two photosystems,as revealed by analysis of the variable-yield fluorescence andthe kinetics of the 520 nm light-induced absorbance change.During the phase of greening at which photosynthetic capacitydeveloped its maximum quantum efficiency, no significant changesin type or content of the various chloroplast cytochromes weredetected. Analysis of the ratio of chlorophyll/plastoquinone,however, showed that changes in this value followed more closelythe observed increase in the quantum efficiency of photosynthesisand the other parameters of photosynthesis examined. (Received May 19, 1972; ) 相似文献
6.
Plants, algae and cyanobacteria grow because of their ability to use sunlight to extract electrons from water. This vital reaction is catalysed by the Photosystem II (PSII) complex, a large multi-subunit pigment-protein complex embedded in the thylakoid membrane. Recent results show that assembly of PSII occurs in a step-wise fashion in defined regions of the membrane system, involves conserved auxiliary factors and is closely coupled to chlorophyll biosynthesis. PSII is also repaired following damage by light. FtsH proteases play an important role in selectively removing damaged proteins from the complex, both in chloroplasts and cyanobacteria, whilst undamaged subunits and pigments are recycled. The chloroplastic Deg proteases play a supplementary role in PSII repair. 相似文献
8.
Picosecond fluorescence kinetics of pea chloroplasts have been investigated at room temperature using a pulse fluorometer with a resolution time of 10 ?11 s. Fluorescence has been excited by both a ruby and neodymium-glass mode-locked laser and has been recorded within the 650 to 800 nm spectral region.We have found three-component kinetics of fluorescence from pea chloroplasts with lifetimes of 80, 300 and 4500 ps, respectively. The observed time dependency of the fluorescence of different components on the functional state of the photosynthetic mechanism as well as their spectra enabled us to conclude that Photosystem I fluoresces with a lifetime of 80 ps ( τI) and Photosystem II fluoresces with a lifetime of 300 ps ( τII). Fluorescence with a lifetime of 4500 ps ( τIII) may be interpreted as originating from chlorophyll monomeric forms which are not involved in photosynthesis.It was determined that the rise time of Photosystem I and Photosystem II fluorescence after 530 nm photoexcitation is 200 ps, which corresponds to the time of energy migration to them from carotenoids. 相似文献
9.
Electron transport from Photosystem II to Photosystem I of spinach chloroplasts can be stimulated by bicarbonate and various carbonyl or carboxyl compounds. Monovalent or divalent cations, which have hitherto been implicated in the energy distribution between the two photosystems, i.e., spillover phenomena at low light intensities, show a similar effect under high light conditions employed in this study. A mechanism for this stimulation of forward electron transport from Photosystem II to Photosystem I could involve inhibition of two types of Photosystem II partial reactions, which may involve cycling of electrons around Photosystem II. One of these is the DCMU-insensitive silicomolybdate reduction, and the other is ferricyanide reduction by Photosystem II at pH 8 in the presence of dibromothymoquinone. Greater stimulation of forward electron transport reactions is observed when both types of Photosystem II cyclic reactions are inhibited by bicarbonate, carbonyl and carboxyl-type compounds, or by certain mono- or divalent cations.Abbreviations used: DCMU, 3-(3,4-dichlorophenyl)-1, 1-dimethylurea; DCIP, 2,6-dichloroindophenol; DBMIB, 2,5-dibromo-3-methyl-6-isopropyl- p-benzoquinone; FeCN, potassium ferricyanide; MV, methylviologen; PS I, photosystem I; PS II, photosystem II; SM, silicomolybdic acid. 相似文献
10.
Stable light-induced absorbance changes in chloroplasts at −196 °C were measured across the visible spectrum from 370 to 730 nm in an effort to find previously undiscovered absorbance changes that could be related to the primary photochemical activity of Photosystem I or Photosystem II. A Photosystem I mediated absorbance increase of a band at 690 nm and a Photosystem II mediated absorbance increase of a band at 683 nm were found. The 690-nm change accompanied the oxidation of P 700 and the 683-nm increase accompanied the reduction of C-550. No Soret band was detected for P 700. A specific effort was made to measure the difference spectrum for the photooxidation of P680 under conditions (chloroplasts frozen to −196 °C in the presence of ferricyanide) where a stable, Photosystem II mediated EPR signal, attributed to P680+ has been reported. The difference spectra, however, did not show that P680+ was stable at −196 °C under any conditions tested. Absorbance measurements induced by saturating flashes at −196 °C (in the presence or absence of ferricyanide) indicated that all of the P680+ formed by the flash was reduced in the dark either by a secondary electron donor or by a backreaction with the primary electron acceptor. We conclude that P680+ is not stable in the dark at −196 °C: if the normal secondary donor at −196 °C is oxidized by ferricyanide prior to freezing, P680+ will oxidize other substances. 相似文献
11.
Most chloroplastic outer envelope membrane proteins are synthesized in the cytosol at their mature size without a cleavable targeting signal. Their insertion into the outer membrane is insensitive to thermolysin pretreatment of chloroplasts and does not require ATP. The insertion has been assumed to be mediated by a spontaneous mechanism or by interaction solely with the lipid components of the outer membrane. However, we show here that insertion of an outer membrane protein requires some trypsin-sensitive and some N-ethylmaleimide-sensitive components of chloroplasts. Association and insertion of the outer membrane protein are saturable and compete with the import of another outer membrane protein. These data suggest that import of chloroplastic outer membrane proteins occurs at specific proteinaceous sites on chloroplasts. 相似文献
12.
Chlorophyll-protein complexes of photosystems (PS), as well as ultrastructural arrangement of chloroplasts in pea leaves of the primary cultivar Torsdag and mutants chlorotica 2004 and 2014 were studied. It was shown that both mutants accumulated 80 and 55% of chlorophyll, respectively, and were able to synthesize all of four types of photosystems complexes. The value of the light-harvesting antenna in mutant 2014 was close to the control one, and in mutant 2004 it increased significantly (by 30%). These changes were caused by a proportional decrease, 40-50%, of any complexes in mutant 2014, whereas the number of PS I reaction centers in mutant 2004 decreased by 50% and the reaction centers of PS II complexes were almost completely retained. It was established that the proportional decrease of PS I and PS II complexes in mutant chlorotica 2014 was followed by a partial reduction of the entire membrane system in chloroplasts, but with a good development of both granal and intergranal sites of thylakoids. On the contrary, the loss of complexes of PS I reaction centers in mutant chlorotica 2004 led to a reduction of unstacked sites of thylakoids in chloroplasts. It was concluded that the disturbance of the lateral orientation of the membrane system of chloroplasts is associated with the loss of complexes of reaction centers of PS I, which is predominantly localized in unstacked sites of thylakoids. 相似文献
13.
A 120 min incubation period with sulfhydryl reagents, such as p-chloromercuribenzoic acid, shows greater than 50% loss of electron-transport activity in Photosystem (PS) II of spinach chloroplasts. Since p-chloromercuriphenylsulfonic acid, a nonpenetrating sulfhydryl reagent, and 4,4′-dithiopyridine, a bifunctional sulfhydryl reagent, show greater inhibition of 3-(3,4-dichlorophenyl)-1,1-dimethylurea-insensitive silicomolybdate reduction than of dibromothymoquinone-insensitive indophenol reduction, it is postulated that two different sulfhydryl reagent-sensitive sites are involved in the PS II electron-transport chain of spinach chloroplasts. 相似文献
14.
The fluorescence yield of chloroplasts reflects the redox state of the electron acceptor of the Photosystem II reaction center, with increasing yield as the acceptor is reduced. Chemical reductive titrations of fluorescence yield in chloroplasts at room temperature indicate two distinct midpoint potentials, suggesting the possibility of Photosystem II electron acceptor heterogeneity. We have carried out a potentiometric titration of the fluorescence decay kinetics in spinach chloroplasts using a continuous mode-locked dye laser with low-intensity excitation pulses and a picosecond-resolution single-photon timing system. At all potentials the fluorescence decay is best described by three exponential components. As the potential is lowered, the slow phase changes 30-fold in yield with two distinct midpoint potentials, accompanied by a modest (3-fold) increase in the lifetime. The titration curve for the slow component of the fluorescence decay of spinach chloroplasts is best characterized by two single-electron redox reactions with midpoint potentials at pH 8.0 of +119 and ?350 mV, with corresponding relative contributions to the fluorescence yield of 49 and 51%, respectively. There is little change in the fast and middle components of the fluorescence decay. We found that the oxidized form of the redox mediator 2-hydroxy-1,4-naphthoquinone preferentially quenches the fluorescence, causing an anomalous decrease in the apparent midpoint of the high-potential transition. This effect accounts for a significant difference between the midpoint potentials that we observe and some of those previously reported. The selective effect of reduction potentials on particular fluorescence decay components provides useful information about the organization and distribution of the Photosystem II electron acceptor. 相似文献
15.
The stoichiometry of the proteins of the photosynthetic oxygen evolution system and of the electron transport components in Photosystem II particles prepared with Triton X-100 from spinach chloroplasts were determined. Per about 220 chlorophyll molecules, there were one reaction center II, one molecule each of the 33, 24 and 18 kDa proteins, four Mn atoms, two cytochromes b-559 (one high-potential, the other low-potential), and 3.5 plastoquinone-9 molecules, but practically no cytochrome b-563, cytochrome f, phylloquinone, α-tocopherol or α-tocopherylquinone. 相似文献
17.
The inhibitory effect of the Cd 2+ in the electron transport of the isolated chloroplasts has been observed by measuring the oxygen uptake from the solution and the fluorescence induction. Cd 2+ is found to be an inhibitor on the donor side of Photosystem II and its action site, as determined by experiments using hydroxylamine and exogenous Mn, is supposed to be on the water-splitting enzyme itself. Moreover, physicochemical and physiological studies indicate that only the ionic form of Cd is acting at the level of the manganoprotein. It is not possible, from this work, to define precisely in which form Cd is taken up through the thylakoid membranes. 相似文献
19.
The photosynthetic activity of the green alga Scenedesmus quadricauda was investigated during synchronous growth in light/dark cycles. The rate of O 2 evolution increased 2-fold during the first 3 to 4 h of the light period, remained high for the next 3 to 4 h, and then declined during the last half of the light period. During cell division, which occurred at the beginning of the dark period, the ability of the cells to evolve O 2 was at a minimum. To determine if photosystem II (PSII) controls the photosynthetic capacity of the cells during the cell cycle we measured PSII activity and heterogeneity. Measurements of electron-transport activity revealed two populations of PSII, active centers that contribute to carbon reduction and inactive centers that do not. Measurements of PSII antenna sizes also revealed two populations, PSII α and PSII β, which differ from one another by their antenna size. During the early light period the photosynthetic capacity of the cells doubled, the O 2-evolving capacity of PSII was nearly constant, the proportion of PSII β centers decreased to nearly zero, and the proportion of inactive PSII centers remained constant. During the period of minimum photosynthetic activity 30% of the PSII centers were insensitive to the inhibitor 3-(3,4-dichlorophenyl)-1,1-dimethylurea, which may be related to reorganization of the thylakoid membrane. We conclude from these results that PSII does not limit the photosynthetic activity of the cells during the first half of the light period. However, the decline in photosynthetic activity observed during the last half of the light period can be accounted for by limited PSII activity. 相似文献
20.
By density gradient centrifugation of the 80000 × g supernatant of digitonintreated spinach chloroplasts two main green bands and one minor green band were obtained. The purification and properties of the particles present in the main bands, which were shown to be derived from Photosystem I and Photosystem II, have been described previously; those of the particles in the minor fraction will be described in the present paper. After purification, these particles show Photosystem II activity but are devoid of Photosystem I activity. They have a high chlorophyll a/chlorophyll b ratio and are enriched in β-carotene and cytochrome b559. At liquid nitrogen temperature, photoreduction of C550 and photooxidation of cytochrome-b559 can be observed. At room temperature, cytochrome b559 undergoes slight photooxidation. These properties indicate that this particle may be the reaction-center complex of Photosystem II. It is suggested that, in vivo, the Photosystem II unit is made up of a reaction-center complex and an accessory complex, the latter being found in one of the main green bands of the density gradient. 相似文献
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