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1.
1. Changes in fluorescence yield of chlorophyll a in isolated chloroplasts have been interpreted by means of regulation of excitation transfer between two pigment systems of photosynthesis 5–7. In order to investigate the relationship between the membrane structure of chloroplasts and the regulation of excitation transfer, changes of light scattering and chlorophyll a fluorescence of isolated spinach chloroplasts were measured upon addition of cations, Mg 2+ and Na +. The cations increased the intensities of both light scattering and fluorescence yield. The changes showed similar time courses and concentration dependences. These facts suggest that modification of membrane structure produced by the cations suppresses the excitation transfer between the two pigment systems. 2. In another case of structural change which is induced by light in the presence of N-methylphenazonium methosulfate, there was little correlation between light-scattering and fluorescence changes. 3. Changes in fluorescence yield induced by the addition of Mg2+ were measured in disintegrated chloroplasts and fractionated particles. The effects of Mg2+ on fluorescence were observed only in preparations of grana stacks, but not in preparations of stroma lamellae. These findings suggest that the excitation transfer is regulated between the two pigment systems located in the grana thylacoid membranes. 相似文献
2.
The effects of monovalent cations on the light energy distribution between two pigment systems of photosynthesis were studied in isolated spinach chloroplasts by measuring chlorophyll a fluorescence and photochemical reactions. The addition of NaCl to the chloroplast suspension produced a 40–80% increase in fluorescence yield measured at 684 nm at room temperature. The fluorescence increase was completed about 5 min after the addition. The effect saturated at 100 mM NaCl. Low-temperature fluorescence spectra showed that NaCl increased the yields of two fluorescence bands of pigment system II at 684 and 695 nm but decreased that of pigment system I at 735 nm. Similar effects on chlorophyll a fluorescence at room and at low temperatures were obtained with NaBr, NaNO3, Na2SO4, LiCl, KCl, RbCl, CsCl, NH4Cl and CH3NH3Cl. NaCl suppressed the quantum efficiency of NADP+ reduction supported by the ascorbate-2,6-dichlorophenolindophenol (DCIP) couple as an electron donor system in the presence of 3-(3′,4′-chlorophenyl)-1,1-dimethylurea (DCMU). On the other hand, NaCl only slightly enhanced the quantum yield of photoreaction II measured by the Hill reaction with DCIP. It is concluded that the monovalent cations tested suppressed the excitation transfer from pigment system II to pigment system I; the effects were the same as those of alkaline earth metals and Mn2+ (refs. 1, 2). 相似文献
3.
Analyses of chlorophyll fluorescence induction kinetics from DCMU-poisoned thylakoids were used to examine the contribution of the light-harvesting chlorophyll a/ b protein complex (LHCP) to Photosystem II (PS II) heterogeneity. Thylakoids excited with 450 nm radiation exhibited fluorescence induction kinetics characteristic of major contributions from both PS II and PS II β centres. On excitation at 550 nm the major contribution was from PS II β centres, that from PS II centres was only minimal. Mg 2+ depletion had negligible effect on the induction kinetics of thylakoids excited with 550 nm radiation, however, as expected, with 450 nm excitation a loss of the PS II component was observed. Thylakoids from a chlorophyll- b-less barley mutant exhibited similar induction kinetics with 450 and 550 nm excitation, which were characteristic of PS II β centres being the major contributors; the PS II contribution was minimal. The fluorescence induction kinetics of wheat thylakoids at two different developmental stages, which exhibited different amounts of thylakoid appression but similar chlorophyll a/ b ratios and thus similar PS II:LHCP ratios, showed no appreciable differences in the relative contributions of PS II and PS II β centres. Mg 2+ depletion had similar effects on the two thylakoid preparations. These data lead to the conclusion that it is the PS II:LHCP ratio, and probably not thylakoid appression, that is the major determinant of the relative contributions of PS II and PS II β to the fluorescence induction kinetics. PS II characteristics are produced by LHCP association with PS II, whereas PS II β characteristic can be generated by either disconnecting LHCP from PS II or by preferentially exciting PS II relative to LHCP. 相似文献
4.
1. Comparative studies were made on the fluorescence characteristics of chlorophyll a at 20° and −193°, and quantum efficiencies for P 700 oxidation and NADP + reduction were measured in chloroplasts and chloroplast fragments obtained after incubation with 0.5% digitonin. 2. Differences in the flurescence yield of chlorophyll a in flowing and stationary suspensions of untreated chloroplasts and of the large fragments are indicative of light-induced photoreduction of the quencher Q of chlorophyll a, associated with pigment System 2 (chlorophyll a2). The relatively low constant fluorescence yield of chlorophyll a in the small fragments indicates the absence of fluorescent chlorophyll a2 from these fragments and suggests that the low fluorescence is due to chlorophyll a, associated with pigmen System 1 (chlorophyll a1). The ratio of the fluorescence yields of chlorophyll a1 and chlorophyll a2 is 0.45:1. In the large particles the concentration ratio of pigment System 1 and System 2 is 1:3. 3. The efficiencies of quanta absorbed at 673, 683 and 705 nm for NADP+ reduction and P 700 oxidation in untreated chloroplasts and chloroplast fragments indicate that digitonin treatment results in a separation of System 2 from System 1 in the small fragments. Sonication does not cause such a separation. Under the conditions used P 700 oxidation and NADP+ reduction in the small fragments separated after digitonin treatment, occurred with maximal efficiency of 0.7 to 1.0 and 0.7, respectively. 4. The constancy of the fluorescence yield of chlorophyll a1 in the small fragments, under conditions at which P 700 is oxidized and NADP+ is reduced, is interpreted as evidence either for the hypothesis that the fluorescence of chlorophyll a1 is controlled by the redox state of the primary photoreductant XH, or alternatively for the hypothesis that energy transfer from fluorescent chlorophyll a1 to P 700 goes via an intrinsically weak fluorescent, still unknown, chlorophyll-like pigment. 5. The low-temperature emission band around 730 nm is argued not to be due to excitation by System 1 only; the relatively large half width of the band, as compared to the emission bands at 683 and 696 nm, suggests that it is possibly due to overlapping emission bands of different pigments. 相似文献
5.
The kinetics of chlorophyll a fluorescence were measured at 685 nm in intact cells of Porphyra yezoensis during alternate illumination of the organism with two colors of light, one absorbed by phycoerythrin and the other by chlorophyll a. Two components of fluorescence change overlapping each other in time were separated; the fast component may be controlled by the rate of Photoreaction II which competes with the fluorescence emission process, and the slow component by the light-induced change in excitation transfer between two pigment systems as suggested in our previous study 6. The kinetics of the slow change in fluorescence yield were extensively investigated. Terms, “State I” and “State II” are used to describe the state of excitation transfer. In the State I a lesser amount of excitation energy is delivered in Pigment System I and greater to Pigment System II than in the State II. The conversion of the states is achieved by the selective illumination of pigment systems. The conversion from the State I toward the State II occurred under Light II (light absorbed by Pigment System II) with a half time of about 10 sec, and it saturated at a light intensity of less than 1000 ergs×cm−2×sec−1. The reverse conversion occurred under Light I (light absorbed by Pigment System I) with a half time of about 5 sec, and it saturated at about 10 000 ergs×cm−2×sec−1. Light I and Light II competed with each other in the interconversion of the states. 相似文献
6.
The role of divalent cations in the regulation of the distribution of excitation energy between the two photosystems involved in green plant photosynthesis has led us to search for a better understanding of how such phenomena might occur at the molecular level. Since small changes in orientation of and distance between pigment molecules could greatly affect the distribution of excitation energy, we have decided to study the effects of ions on the light-harvesting pigment protein from spinach chloroplasts. The light-harvesting pigment protein is shown to have two types of binding sites for Ca 2+. Binding studies and analytical ultracentrifugation indicate that site I ( Kd = 2.5 μM, n = 1.5−4.0 μmol Ca 2+ bound/mg chlorophyll) is lost as the protein associates. Site II ( Kd = 32 μM, n = 9.5 μmol Ca 2+/mg chlorophyll) is not affected by the association of the protein. This site is responsible, however, for a further divalent cation-dependent association of the protein. The possible role of this protein in grana stacking and control of spillover is discussed. 相似文献
7.
The formation of the triplet state of carotenoids (detected by an absorption peak at 515 nm) and the photo-oxidation of the primary donor of Photosystem II, P-680 (detected by an absorption increase at 820 nm) have been measured by flash absorption spectroscopy in chloroplasts in which the oxygen evolution was inhibited by treatment with Tris. The amount of each transient form has been followed versus excitation flash intensity (at 590 or 694 nm). At low excitation energy the quantum yield of triplet formation (with the Photosystem II reaction center in the state Q −) is about 30% that of P-680 photo-oxidation. The yield of carotenoid triplet formation is higher in the state Q − than in the state Q, in nearly the same proportion as chlorophyll a fluorescence. It is concluded that, for excited chlorophyll a, the relative rates of intersystem crossing to the triplet state and of fluorescence emission are the same in vivo as in organic solvent. At high flash intensity the signal of P-680 + completely saturates, whereas that of carotenoid triplet continues to increase. The rate of triplet-triplet energy transfer from chlorophyll a to carotenoids has been derived from the rise time of the absorption change at 515 nm, in chloroplasts and in several light-harvesting pigment-protein complexes. In all cases the rate is very high, around 8 · 107 s−1 at 294 K. It is about 2–3 times slower at 5 K. The transitory formation of chlorophyll triplet has been verified in two pigment-protein complexes, at 5 K. 相似文献
8.
Ni 2+ inhibits electron-transport activity of isolated barley chloroplasts and this inhibition of electron transport by Ni 2+ is distinctly different from other heavy metal ion (e.g., Pb 2+, Cd 2+, Zn 2+)-induced inhibition of chloroplast function. Ni 2+ inactivates Photosystem II (PS II) activity at a lower concentration than that required for the same extent of inhibition of Photosystem I (PS I)-mediated electron flow. Ni 2+ induces changes in chlorophyll a (Chl a) emission characteristics and brings about a lowering of the Chl a fluorescence yield, and this lowering of Chl a fluorescence intensity is not relieved by the exogenously supplied electron donor NH 2OH which donates electrons very close to the PS II reaction centres. Immobilization of the chloroplast membrane structure with glutaraldehyde fails to arrest the Ni 2+-induced loss of PS II activity. Also, Ni 2+-treated chloroplasts do not regain the ability to photoreduce 2,6-dichlorophenolindophenol even after washing of chloroplasts with buffer. These results indicate that unlike Zn 2+ or Pb 2+, Ni 2+ induces alterations in the chloroplast photosynthetic apparatus resulting in an irreversible loss of electron-transport activity. 相似文献
9.
The fast fluorescence decay kinetics of two photosynthetic mutants of corn ( Zea mays) have been compared with those of normal corn. The fluorescence of normal corn can be resolved into three exponential decay components of lifetime 900–1500 ps (slow), 300–500 ps (middle) and 50–120 ps (fast), the yields of which are affected by light intensity and Mg 2+ levels. The Photosystem II-(PS II)-defective mutant hcf-3 has similar decay lifetimes (approx. 1200, 450 and 100 ps) but is not affected by light intensity, reflecting the absence of PS II charge recombination. However, yields do respond to Mg 2+ in a fashion typical of normal corn, which may be correlated with the presence of normal levels of light-harvesting chlorophyll a + b complex (LHCP). The PS I mutant hcf-50 also shows three-component decay kinetics. In conjunction with the results on the LHCP-deficient mutant of barley presented in a recent paper (Karukstis, K.K. and Sauer, K. (1984) Biochim. Biophys. Acta 766, 148–155), these data suggest that the slow component of normal chloroplasts is kinetically controlled by the decay processes of the LHCP and that the energy comes from one of two sources: (a) charge recombination in the reaction centre or (b) energy transferred within or between LHCP units only. The fast component appears to originate from both PS I and PS II. The complex response of the middle component to cations and light intensity, and its presence in all of the mutants, suggests that it also may have multiple origins. 相似文献
10.
The metallochromic indicator Eriochrome Blue SE was used to measure light-induced internal movement of Mg 2+ in intact chloroplasts. By dual-wavelength spectroscopy (measuring wavelength 554 nm, reference 592 nm) a light-induced, dark-reversible absorbance increase of Eriochrome Blue in samples of isolated intact chloroplasts was observed. The light/dark difference spectrum of Eriochrome Blue between 550 and 590 nm (reference wavelength 562 nm) indicated that this absorbance increase was caused by an increased concentration of free Mg 2+ in a neutral or slightly alkaline chloroplast compartment. The signal was seen only with intact, but not with broken, envelope-free chloroplasts, which had lost most of their divalent cations. This is interpreted to show that the indicator responds to an increase of Mg2+ concentration in the chloroplast stroma, which represents an efflux of Mg2+ from the intra-thylakoid space caused by light-dependent proton pumping. As calculated from corrected values of the absorbance increase of Eriochrome Blue, the light-induced internal release of Mg2+ was close to 100 nequiv per mg chlorophyll at pH 7.6 and 250 nequiv at pH 7.1. This corresponds to a light-dependent increase in the concentration of free Mg2+ in the stroma of about 2 and 5 mM, respectively. 相似文献
11.
(1) Light-dependent changes of the Mg 2+ content of thylakoid membranes were measured at pH 8.0 and compared with earlier measurements at pH 6.6. In a NaCl and KCl medium, the light-dependent decrease in the Mg 2+ content of the thylakoid membranes at pH 8.0 is found to be 23 nmol Mg 2+ per mg chlorophyll, whereas in a sorbitol medium it is 83 nmol Mg 2+ per mg chlorophyll. (2) A light dependent increase in the Mg2+ content of the stroma was detected when chloroplasts were subjected to osmotic shock, amounting to 26 nmol/mg chlorophyll. Furthermore, a rapid and reversible light-dependent efflux of Mg2+ has been observed in intact chloroplasts when the divalent cation ionophore A 23 187 was added, indicating a light-dependent transfer of about 60 nmol of Mg2+ per mg chlorophyll from the thylakoid membranes to the stroma. (3) CO2 fixation, but not phosphoglycerate reduction, could be completely inhibited when A 23 187 was added to intact chloroplasts in the absence of external Mg2+. If Mg2+ was then added to the medium, CO2 fixation was restored. Half of the maximal restoration was achieved with about 0.2 mM Mg2+, which is calculated to reflect a Mg2+ concentration in the stroma of 1.2 mM. The further addition of Ca2+ strongly inhibits CO2 fixation. (4) The results suggest that illumination of intact chloroplasts causes an increase in the Mg2+ concentration of 1–3 mM in the stroma. Compared to the total Mg2+ content of chloroplasts, this increase is very low, but it appears to be high enough to have a possible function in the light regulation of CO2 fixation. 相似文献
12.
The following arguments in favor of exciton transfer between the two photosystems are presented: 1. (1) MgCl2 (1–10 mM range) decreases the intersystem transfer but does not modify the partition of absorbed photons between the photosystems. MgCl2 addition causes a simultaneous increase of excitation life time (τ) and of fluorescence intensity (F). The same linear relationship is obtained with or without added Mg2+. 2. (2) The deactivation of Photosystem II by the Photosystem II to Photosystem I transfer increases with the level of reduced Photosystem II traps. When all Photosystem II traps are closed, half of Photosystem II excitons are deactivated by transfer to Photosystem I. 3. (3) From the relative values of the 685-nm fluorescence yield and System II electron transport rate in limiting light, measured with and without MgCl2, the values of rate constants of Photosystem II deactivation were calculated. 4. (4) The intersystem transfer determines a 715-nm variable fluorescence, which is lowered by MgCl2 addition. When this transfer is decreased by MgCl2 the efficiency of the transfer between Photosystem II-connected units is enhanced, and a more sigmoidal fluorescence rise is obtained.
A double-layer model of the thylakoid membrane where each photosystem is restricted to one leaflet is proposed to explain the decrease of the intersystem transfer after adding cations. It is suggested that MgCl2 decreases the thickness of the Photosystem I polar region, increasing the distance between the pigments of the two photosystems. 相似文献
13.
The kinetics of fluorescence yield in Chlorella pyrenoidosa and spinach chloroplasts were studied in the time range of 0.5 μs to several hundreds of microseconds in the presence of hydroxylamine. Fluorescence was excited with a just-saturating xenon flash with a halfwidth of 13 μs (λ = 420 nm). The fast rise of the fluorescence yield which was limited by the rate of light influx, was, in the presence of 10 −3–10 −2 M hydroxylamine, replaced by a slow component which had a half risetime of 25 μs in essence independent of light intensity. This slow fluorescence yield increase reflects a dark reaction on the watersplitting side of Photosystem II. Simultaneous oxygen evolution measurements suggested that a fast fluorescence component is only present in organisms with intact O 2-evolving system, whereas a slow rise predominantly occurs in organisms with the watersplitting system irreversibly inhibited by hydroxylamine. The results can be explained by the following hypotheses: (a) The primary donor of Photosystem II in its oxidized state, P+, is a fluorescence quencher. (b) Hydroxylamine prevents the secondary electron donor Z from reducing the oxidized reaction center pigment P+ rapidly. This inhibition is dependent on hydroxylamine concentration and is complete at a concentration of 10−2 M. (c) A second donor (not transporting electrons from water) transfers electrons to P+ with a half time of roughly 25 μs. 相似文献
14.
1. 1. The effect of the Mg2+ concentration on the CO2 fixation activity in situ in isolated and intact spinach chloroplasts upon suspension in hypotonic medium was examined. CO2 fixation in the dark was activated 25–100 fold by 20 mM Mg2+ in the presence of added ATP plus either ribulose 5-phosphate or ribose 5-phosphate. 20 mM Mg2+-stimulated fixation only 2–3 fold in the presence of the substrate of fixation, ribulose 1,5-diphosphate. The highest Mg2+-stimulated rate of fixation in the dark observed with chloroplasts was 480 μmoles CO2 fixed per mg chlorophyll per h. 2. 2. The concentration of bicarbonate at half of the maximal velocity (apparent Km) during the Mg2+-stimulated fixation of CO2 was 0.4 mM in the presence of ATP plus ribose 5-phosphate and 0.6 mM with ribulose 1,5-diphosphate. 3. 3. Dithioerythritol or light enhanced Mg2+-stimulated CO2 fixation 1–3 fold in the presence of ATP plus ribose 5-phosphate but not ribulose 1,5-diphosphate. 4. 4. These results indicate that Mg2+ fluxes in the stroma of the chloroplast could control the activity of the phosphoribulokinase with a lesser effect on the ribulosediphosphate carboxylase. An increase in Mg2+ of 6–10 mM in the stroma region of the chloroplast would be enough to activate CO2 fixation during photosynthesis.
Abbreviations: Rib-5-P, ribose 5-phosphate; Ribul-5-P, ribulose 5-phosphate; Ribul-1,5-P2, ribulose 1,5-diphosphate; HEPES, N-2-hydroxyethylpiperazine-N′-2-ethanesulfonic acid; MES, 2-(N-morpholino)ethanesulfonic acid 相似文献
15.
The effects of Mn 2+ on aerobic photobleaching of carotenoids, on photoreduction of 2,6-dichlorophenolindophenol (DCIP) and on fluorescence above 600 mμ of spinach chloroplasts washed with 0.8 M Tris-HC1 buffer were investigated. Carotenoids (mostly carotenes, lutein and violaxanthin) in the Tris-washed chloroplasts were irreversibly bleached by illumination with red light, while carotenoids in normal chloroplasts prepared with a low concentration of Tris-HC1 underwent no bleaching upon illumination. The photobleaching of carotenoids observed with Tris-washed chloroplasts was inhibited by Mn 2+ (MnCl 2 or MnSO 4) as well as by some inhibitors of the Hill reaction such as dichlorophenyl-1,1-dimethylurea (DCMU), methylthio-4,6-bis-isopropylamino- s-triazine and o-phenanthroline or by reducing agents such as ascorbate plus tetramethyl- p-phenylene diamine (TMPD). DCIP photoreduction, which was deactivated by Tris, was reactivated to 50–80% of the rate for normal chloroplasts upon addition of Mn 2+. The restored photoreduction of DCIP was inhibited by DCMU and carbonylcyanide m-chlorophenylhydrazone (CCCP). The steady-state fluorescence yield of normal chloroplasts measured at room temperature was lowered by Tris treatment, and the decreased yield was restored by adding Mn 2+ as well as ascorbate plus TMPD. CCCP also lowered the yield; the yield was recovered by adding ascorbate plus TMPD. Determination of manganese in normal and Tris-washed chloroplasts showed that 30% of the manganese in chloroplast was removed with Tris. It was postulated that Mn 2+ functions in the electron transport on the oxidizing side of Photosystem II at a site between water and an electron carrier ( Y). CCCP as well as Tris inhibits the reduction of Y+ by Mn 2+, and carotenoids are oxidized by Y+ which is reduced by ascorbate plus TMPD. 相似文献
16.
Mg 2+ can potentially influence bacterial adhesion directly through effects on electrostatic interactions and indirectly by affecting physiology-dependent attachment processes. However, the effects of Mg 2+ on biofilm structure are largely unknown. In this study, Pseudomonas fluorescens was used to investigate the influence of Mg 2+ concentration (0, 0.1 and 1.0 mM MgCl 2) on biofilm growth. Planktonic and attached cells were enumerated (based on DAPI staining) while biofilm structures were examined via confocal laser scanning microscopy and three-dimensional structures were reconstructed. Mg 2+ concentration had no influence on growth of planktonic cells but, during biofilm formation, Mg 2+ increased the abundance of attached cells. For attached cells, the influence of Mg 2+ concentration changed over time, suggesting that the role of Mg 2+ in bacterial attachment is complex and dynamic. Biofilm structures were heterogeneous and surface colonization and depth increased with increasing Mg 2+ concentrations. Overall, for P. fluorescens, Mg 2+ increased initial attachment and altered subsequent biofilm formation and structure. 相似文献
17.
The kinetics of LHCP phosphorylation and associated changes in photosystem cross-section and energy ‘spill-over’ from PS II to PS I have been examined in isolated spinach chloroplasts. During an initial phosphorylation period of 3–6 min, in the presence of saturating concentrations of Mg 2+, the increase in PS I and decrease in PS II cross-section are largely completed, as judged by both measurements of the steady-state redox state of Q and fluorescence yield changes. This corresponds to a period of rapid 32P incorporation into the low-molecular weight LHCP polypeptide. Subsequent to this initial 3–6-min period there is substantial further phosphorylation of both LHCP polypeptides, which is not accompanied by significant changes in photosystem cross-section, even after the chloroplasts had been unstacked with extensive mixing of PS I and PS II by Mg-removal. It is suggested that there exists a specific ‘mobile’ population of LHCP molecules which is rapidly phosphorylated and which may be enriched in the low-molecular-weight polypeptide. In addition, measurements of the kinetics of the ‘spill-over’ changes upon either Mg 2+ addition or removal indicate that the continued phosphorylation of LHCP is able to increase the ‘spill-over’ process under favourable ionic conditions. 相似文献
18.
The degree of fluoresence polarization, P, of unoriented and magnetically oriented spinach chloroplasts as a function of excitation (400–680 nm) and emission wavelengths (675–750 nm) is reported. For unoriented chloroplasts P can be divided into two contributions, PIN and PAN. The latter arises from the optical anisotropy of the membranes which is due to the orientation with respect to the membrane plane of pigment molecules in vivo. The intrinsic polarization PIN, which reflects the energy transfer between different pigment molecules and their degree of mutual orientation, can be measured unambiguously only if (1) oriented membranes are used and the fluorescence is viewed along a direction normal to the membrane planes, and (2) the excitation is confined to the Qy (≈ 660−680 nm) absorption band of chlorophyll in vivo. With 670–680 nm excitation, values of P using unoriented chloroplasts can be as high as +14%, mostly reflecting the orientational anisotropy of the pigments. Using oriented chloroplasts, PIN is shown to be +5±1%. The excitation wavelength dependence studies of PIN indicate that the carotenoid and chlorophyll Qy transition moments tend to be partially oriented with respect to each other on a local level (within a given photosynthetic unit or its immediate neighbors). 相似文献
19.
From studies of electron-transport reactions of isolated spinach chloroplasts, we observe the following quantum requirements: (A) For the photoreduction of NADP +, measured both aerobically and anaerobically, in a 3-(3,4-dichlorophenyl)-1,1-dimethyl urea (DCMU) poisoned system with ascorbate and reduced 2,6-dichlorophenolindophenol (DCIPH 2) present as electron donors, the quantum requirements are 1.0 ± 0.05 at wavelengths longer than 700 nm of actinic light, and 1.5–2.5 for wavelengths between 620 and 680 nm. (B) For the photoreduction of 2,6-dichlorophenolindophenol (DCIP) with water as the electron donor, the quantum requirements are 1.0 ± 0.05 in the range 630–660 nm. (C) For the photoreduction of NADP + with water as the electron donor, the quantum requirements are 2.0 ± 0.1 in the wavelength range 640–678 nm of actinic light, increasing to 6 or greater at wavelengths beyond 700 nm. These results are shown to be inconsistent with the “separate package” model for the two pigment systems in higher plant photosynthetic electron transport. The evidence is most easily interpreted using a “controlled spillover” model, in which the transfer of electronic excitation energy from one pigment system to the other is under the control of incompletely identified factors in the reaction mixture. At moderate light intensities the steady state rate of the [ascorbate + DCIPH2 → NADP+] reaction (A) in the presence of DCMU and added ferredoxin can be increased more than 3 times when saturating amounts of plastocyanin and ferredoxin-NADP reductase are added to the chloroplasts. Similarly, the steady-state rate of the [H2O → DCIP] Hill reaction (B) is increased about 3-fold by added MgCl2 and plastocyanin, but added ferredoxin or ferredoxin-NADP reductase have no effect on this reaction. Plastocyanin appears to be the electron transport component which couples to DCIP, either in the oxidized or in the reduced form, in the reaction media. The steady-state rate of the [H2O → NADP+] reaction (C) with saturating amounts of ferredoxin can be further increased more than 3-fold when MgCl2, plastocyanin and ferredoxin-NADP reductase are added. 相似文献
20.
1. The curves representing the reciprocal fluorescence yield of chlorophyll a of Photosystem II (PS II) in Chlorella vulgaris as a function of the concentration of m-dinitrobenzene in the states P Q and P Q -, are found to be straight parallel lines; P is the primary donor and Q the primary acceptor of PS II. In the weakly trapping state P Q - the half-quenching of dinitrobenzene is about 0.2 mM, in vitro it is of the order of 10 mM. The fluorescence yield as a function of the concentration of a quencher is described for three models for the structure of pigment systems: the model of separate units, the model of limited energy transfer between the units, and the matrix model. If it is assumed that the rate constant of quenching by dinitrobenzene is high and thus the number of dinitrobenzene molecules per reaction center low, it can be concluded that the pigment system of PS II in C. vulgaris is a matrix of chlorophyll molecules in which the reaction centers are embedded. Theoretical and experimental evidence is consistent with such an assumption. For Cyanidium caldarium the zero fluorescence yield Ф0 and its quenching by dinitrobenzene were found to be much smaller than the corresponding quantities for C. vulgaris. Nevertheless, our measurements on C. caldarium could be interpreted by the assumption that the essential properties (rate constants, dinitrobenzene quenching) of PS II are the same for these two species belonging to such widely different groups. 2. The measured dinitrobenzene concentrations required for half-quenching in vivo and other observations are explained by (non-rate-limiting) energy transfer between the chlorophyll a molecules of PS II and by the assumptions that dinitrobenzene is approximately distributed at random in the membrane and does not diffuse during excitation. 3. The fluorescence kinetics of C. vulgaris during a 350 ns laser flash of variable intensity could be simulated on a computer using the matrix model. From the observed fluorescence quenching by the carotenoid triplet (CT) and the measurement of the number of CT per reaction center via difference absorption spectroscopy, the rate constant for quenching of CT is calculated to be kT = 3.3 · 1011 s−1 which is almost equal to the rate constant of trapping by an open reaction center (Duysens, L.N.M. (1979) CIBA Foundation Symposium 61 (New Series), pp. 323–340). 4. The fluorescence quenching by CT in non-treated spinach chloroplasts after a 500 ns laser flash (Breton, J., Geacintov, N.E. and Swenberg, C.E. (1979) Biochim. Biophys. Acta 548, 616–635) could be explained within the framework of the matrix model when the value for kT is used as given in point 3. 5. The observations mentioned under point 1 indicate that the fluorescence yield Ф0 for centers in trapping state P Q is probably for a fraction exceeding 0.8 emitted by PS II. 相似文献
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