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1.
Electric fields of a few hundred volts per centimeter greatly stimulate the emission of delayed light from “broken” chloroplasts. At low intensities of exciting light the fluorescence of these chloroplasts is also stimulated by the electric field, but to a lesser extent. Assuming that the electric field has no effect on prompt fluorescence, and has the same effect on the delayed light emission during illumination as in the dark, we can determine the ratio of delayed light to fluorescence under steady-state illumination. 相似文献
2.
The effects of varying dark interval on the kinetics of light-induced formation of the membrane potential were studied on individual chloroplasts of Anthoceros with the use of capillary microelectrodes. Illumination of the chloroplast with 1 s light pulse after 3 min dark period induced the photoelectrical response with two peaks of the potential that were located at 20 and 500 ms after the onset of illumination. The position of the second peak was shifted along the time-scale depending on the preceding dark interval. The repeated illumination of the chloroplast with 1 s light pulse after 30 s dark interval induced the electrical response with only one maximum and a monotonous decay of the potential in the light. Distinctions in the electrical responses induced by the first and the second light pulses were eliminated by the addition of 50 μM dicyclohexylcarbodiimide (DCCD). The results show that the photoinduction kinetics of the membrane potential in chloroplasts is affected by functioning of H +-ATPase. The delayed peak of the membrane potential in the photoinduction kinetics is interpreted as a consequence of the photoactivated electron transport supported by Photosystem I. 相似文献
3.
KCl-induced luminescence in relation to slow delayed light emission (> 3 s) and pH shift-triggered luminescence was studied in preilluminated chloroplasts. An activation pathway for KCl-induced luminescence similar to that for acid-base-triggered luminescence but different from that for delayed light emission is suggested.When the chloroplasts were subjected to a small amount of pH transition together with a simultaneous addition of KCl, a synergistic enhancement of triggered luminescence was observed. The synergism was not observed when the pH transition was increased. The results are interpreted according to the protonation model for stimulated luminescence. 相似文献
4.
An electric field pulse was applied to a suspension of osmotically swollen spinach chloroplasts after illumination with a saturating flash in the presence of DCMU. In addition to the stimulation of delayed fluorescence by the electric field, discovered by Arnold and Azzi (Arnold, W.A. and Azzi, R. (1971) Photochem. Photobiol. 14, 233–240) a sudden drop in fluorescence yield was observed. The kinetics of this fluorescence change were identical to those of the integrated delayed fluorescence emission induced by the pulse. The S-state dependence of the stimulated emission was very similar to that of the normal luminescence. We assume that the membrane potential generated by the pulse changes the activation energy for the back reaction in Photosystem II. On this basis, and making use of data we obtained earlier from electrochromic absorbance changes induced by the pulse, the kinetics of the field-induced prompt and delayed fluorescence changes, and also the amplitude of the fluorescence decrease, which was about 12% for a nearly saturating pulse, are explained. Our results indicate that in those reaction centers where a decrease of the activation energy occurs the effect of a pulse can be quite spectacular: the back reaction, which normally takes seconds, is completed in a few hundred microseconds when a sufficiently strong pulse is applied. Measurements of the polarization of the stimulated luminescence supported the interpretation given above.Only 2.8% of the back reaction was found to proceed via transition of reexcited chlorophyll to the ground state, both during the field pulse and in the absence of the field. 相似文献
5.
A modified fluorescence microscope system was used to measure chlorophyll fluorescence and delayed light emission from mesophyll and bundle sheath cells in situ in fresh-cut sections from leaves of Panicum miliaceum L. The fluorescence rise in 3-(3,4-dichlorophenyl)-1, 1-dimethylurea (DCMU)-treated leaves and the slow fluorescence kinetics in untreated leaves show that mesophyll chloroplasts have larger photosystem II unit sizes than do bundle sheath chloroplasts. The larger photosystem II units imply more efficient noncyclic electron transport in mesophyll chloroplasts. Quenching of slow fluorescence also differs between the cell types with mesophyll chloroplasts showing complex kinetics and bundle sheath chloroplasts showing a relatively simple decline. Properties of the photosynthetic system were also investigated in leaves from plants grown in soil containing elevated NaCl levels. As judged by changes in both fluorescence kinetics in DCMU-treated leaves and delayed light emission in leaves not exposed to DCMU, salinity altered photosystem II in bundle sheath cells but not in mesophyll cells. This result may indicate different ionic distributions in the two cell types or, alternatively, different responses of the two chloroplast types to environmental change. 相似文献
6.
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
7.
Induction curves of the delayed light emission in spinach chloroplasts were studied by measuring the decay kinetics after each flash of light. This study differs from previous measurements of the induction curves where only the intensities at one set time after each flash of light were recorded. From the decay kinetics after each flash of light, the induction curves of the delayed light emission measured 2 ms after a flash of light were separated into two components: one component due to the last flash only and one component due to all previous flashes before the last one. On comparing the delayed light induction curves of the two components with the fluorescence induction curves in chloroplasts treated with 3-(3,4-dichlorophenyl)-1,1-dimethylurea and in chloroplasts treated with hydroxylamine and 3-(3,4-dichlorophenyl)-1,1-dimethylurea, the component due to the last flash only is found to be dependent on the concentration of open reaction centers and the component due to all previous flashes except the last is dependent on the concentration of closed reaction centers. This implies that the yield of the fast decaying component of the delayed light emission is dependent on the concentration of open reaction centers and the yield of the slow decaying component is dependent on the concentration of closed reaction centers. 相似文献
8.
KCL-induced luminescence in relation to slow delayed light emission (greater than 3 s) and pH shift-triggered luminescence was studied in preilluminated chloroplasts. An activation pathway for KCl-induced luminescence similar to that for acidbase-triggered luminescence but different from that for delayed light emission is suggested. When the chloroplasts were subjected to a small amount of pH transition together with a simultaneous addition of KCl, a synergistic enhancement of triggered luminescence was observed. The synergism was not observed when the pH transition was increased. The results are interpreted according to the protonation model for stimulated luminescence. 相似文献
9.
Yeast cells of Candida albicans which had been attached to polylysine-coated microscope slides were induced to form buds or germ tubes in the presence of external electrical fields. The sites of budding and germ tube formation and the growth of germ tubes and hyphal branches were polarized preferentially towards the cathode. Buds were not converted to pseudohyphae or germ tubes by the field and the field had no effect on the positioning of nuclei or septa in the yeast cell or germ tube. Buds were less polarized than germ tubes at any given applied voltage. The polarization of buds reached a peak at an electrical field of 12 mV per cell. Polarization of germ tubes was biphasic, increasing rapidly with increasing field strengths up to 5 mV per cell, and then more slowly in stronger fields. An electrical field was only required for a fraction of the time taken for germ tubes to start to form, so cells retained a memory of experiencing an electrical field which influenced the selection of sites of evagination. Increasing the electrical field delayed the time of germ tube evagination and inhibited the rate of germ tube extension. Unlike previous findings with other filamentous fungi, germ tubes grew unidirectionally towards the cathode for extended periods and did not deviate to a perpendicular orientation. This result suggests that the septal pore of the filamentous form may have high electrical resistance and would act as an effective barrier to solute transport between intercalary compartments. 相似文献
10.
The cell structure of Chlamydomonas reinhardi is disrupted by brief exposure to sonication. The extent of cell breakage can be determined quickly by cell count with the light microscope. Rates of photochemical activities of briefly sonicated cells approach those reported for higher plant chloroplasts. These activities are a sensitive function of time of sonication and sonic power used. The method of brief sonication is rapid and convenient and gives a stable preparation useful for determining photochemical activities in Chlamydomonas. 相似文献
11.
1. Delayed light of chlorophyll emitted at 0.1–3.9 ms after cessation of repetitive flash light was studied at temperatures between +40 and −196 °C in isolated spinach chloroplasts. 2. Induction kinetics of delayed light varied depending on temperature. It was found to be composed of two phases; one was an initial rapid rise followed by a rather fast decline to a low steady state level (fast phase), and the other was a slow increase after the initial rapid rise to the maximum followed by an insignificant slow decrease to a high steady state level (slow phase). The fast phase existed between −175 and 40 °C with the maximum at −40 °C, while the slow phase, between 0 and 40 °C with the maximum at 25 °C. 3. The intensity of delayed light at −175 °C was found to be less than one fiftieth that at 0 °C, and no delayed light emission was observed at −196 °C within experimental accuracy. This is in contrast to the results reported by Tollin, G., Fujimori, E. and Calvin, M. ((1958) Proc. Natl. Acad. Sci. U.S. 44, 1035–1047) in which the intensity of delayed light measured at −170 °C was about a half that at 0 °C. 4. The induction of delayed light measured at −96 °C was found to be significantly suppressed by the preillumination at −196 °C. This finding suggests that the primary photochemical event still survives at −196 °C without emission of delayed light. 5. Decay kinetics of delayed light after the flash excitation revealed the presence of at least two decay components. A slow decay component with a half decay time of several tens of milliseconds was observed at temperatures higher than 0 °C. A fast decay component with a half decay time of about 0.2 ms was observed at temperatures between −120 and 25 °C. The decay rate of this component was slightly retarded on cooling. 6. The System II particles derived from spinach chloroplasts with digitonin treatment showed a temperature dependence of delayed light similar to that of the chloroplasts. System I particles, on the other hand, scarcely emitted the delayed light at any temperature between 40 and −196 °C. 相似文献
12.
1. Using single chloroplasts of Peperomia metallica the kinetics of light-induced potential changes were studied. Three kinetic components (the initial fast rise, the decay in the light and the decay in the dark) were found to be characterized by time constants 4, 220 and 60 ms, respectively at light intensity 5000 lx and temperature 18 °C. After flash excitation the potential kept on rising for about 10 ms. Cooling of the medium down to 5 °C had no effect on the duration of potential rise after the flash.2. Variations in the medium temperature in the range 2–23 °C had little effect on photoresponse magnitude but resulted in significant acceleration of decay in the light.3. Addition of 3-(3,4-dichlorophenyl)-1,1-dimethylurea (5 · 10 ?6 M) resulted in suppression of the magnitude of the photoresponse but was not accompanied by any change in the rate of initial rise of potential. 3-(3,4-Dichlorophenyl)-1,1-dimethylurea-inhibited photoresponse could be restored and even enhanced by subsequent addition of N-methylphenazonium methosulfate (10 ?4 M). N-Methylphenazonium methosulfate essentially influenced the time course and light-intensity curves of photoresponse.4. The chloroplast photoresponses were of different time-courses when elicited by red (640 nm) or far red (712 nm) light. This fact as well as an enhancement effect of combined illumination by two intermittent light beams indicate on the interaction of two photosynthetic pigment systems when the photoelectric response was formed.5. An imposed electrical field resulted in stimulation or suppression of chloroplast photoresponse depending on the polarity of the field. No indications for the existance of “reversal potential” for photoelectric response were obtained.6. A kinetic scheme of photoresponse formation is proposed, which includes two sequential photochemical reactions of photosynthesis. 相似文献
13.
Effective ionophore:chlorophyll ratios were determined for various ionophores that decrease the electrical potential across thylakoid membranes in intact and hypo-osmotically lysed chloroplasts isolated from spinach ( Spinacia oleracea). The efficacy of gramicidin D, valinomycin, carbonylcyanide m-chlorophenylhydrazone, and dicyclohexano-18-crown-6 in collapsing the electrical potential was determined spectrophotometrically by the decay half-time of the absorbance change at 518 nanometers induced by a saturating, single turnover flash. The results show that the effectiveness of the ionophores in collapsing the electrical potential in intact and lysed chloroplasts depends on the amount of ionophore-accessible membrane in the assay medium. Only gramicidin exhibited a significant difference in efficacy between intact and lysed chloroplasts. The ratio of gramicidin to chlorophyll required to collapse the electrical potential was more than 50 times higher in intact chloroplasts than in lysed chloroplasts. The efficacy of carbonylcyanide m-chlorophenylhydrazone was significantly reduced in the presence of bovine serum albumin. The other ionophores tested maintained their potency in the presence of bovine serum albumin. Valinomycin was the most effective ionophore tested for collapsing the electrical potential in intact chloroplasts, whereas gramicidin was the most potent ionophore in lysed chloroplasts. The significance of the ionophore:chlorophyll ratios required to collapse the electrical potential is discussed with regard to bioenergetic studies, especially those that examine the contribution of the transmembrane electrochemical potential to protein transport into chloroplasts. 相似文献
14.
This paper gives an overview of behavioral studies on the color and polarization vision of the Japanese yellow swallowtail butterfly, Papilio xuthus. We focus on indoor experiments on foraging individuals. Butterflies trained to visit a disk of certain color correctly select that color among various other colors and/or shades of gray. Correct selection persists under colored illumination, but is systematically shifted by background colors, indicating color constancy and simultaneous color contrast. While their eyes contain six classes of spectral receptors, their wavelength discrimination performance indicates that their color vision is tetrachromatic. P. xuthus innately prefers brighter targets, but can be trained to select dimmer ones under certain conditions. Butterflies trained to a dark red stimulus select an orange disk presented on a bright gray background over one on dark gray. The former probably appears darker to them, indicating brightness contrast. P. xuthus has a strong innate preference for vertically polarized light, but the selection of polarized light changes depending on the intensity of simultaneously presented unpolarized light. Discrimination of polarization also depends on background intensity. Similarities between brightness and polarization vision suggest that P. xuthus perceive polarization angle as brightness, such that vertical polarization appears brighter than horizontal polarization. 相似文献
15.
The addition of the cyclic cofactor 2,3,5,6-tetramethyl- p-phenylenediamine (diaminodurene) to a suspension of chromatophores of Rhodopseudomonas spheroides causes a light-dependent quenching of bacteriochlorophyll fluorescence. This effect is similar to one observed in chloroplasts and related to proton uptake. It is distinct from the quenching operative through the redox state of the primary electron donor and acceptor, as shown by its sensitivity to uncouplers and ionophorous antibiotics. The quenching is dependent on light intensity and diaminodurene concentration, and has a pH optimum at 7.1 where up to 70% of the fluorescence could be quenched in the presence of 0.33 mM diaminodurene. 相似文献
16.
Delayed light emission from the Triton-fractionated Photosystem II subchloroplast fragments (TSF-IIa) was measured between 0.5 and 10 ms after the termination of illumination. The delayed light emission was diminished by Photosystem II inhibitors, DCMU and o-phenanthroline, which act between the reduced primary acceptor and the plastoquinone pool.Secondary electron donors to Photosystem II, diphenylcarbazide, phenylenediamine, Mn 2+, and ascorbate inhibited delayed light emission. Secondary electron acceptors such as ferricyanide, dichlorophenol indophenol, and dimethyl benzoquinone enhanced delayed light emission. The addition of secondary electron acceptors to TSF-IIa particles containing Mn 2+ restored delayed light emission to almost the control level. The plastoquinone antagonist, 2,5-dibromo-3-methyl-6-isopropyl p-benzoquinone, increased delayed light emission at low concentrations but decreased it at higher concentrations. Silicomolybdate enhanced the delayed light emission of TSF-IIa particles markedly, and reversed the inhibition by DCMU. Silicomolybdate showed a similar stimulatory effect on the delayed-light intensity in broken spinach chloroplasts at shorter times after the termination of illumination. Carbonyl cyanide m-chloro (or p-trifluoromethoxy) phenylhydrazones inhibited the delayed light emission, but NH 4Cl had no effect. 相似文献
17.
The effect of 2,3,5,6-tetramethyl p-phenylenediamine-catalyzed cyclic electron flow on millisecond delayed light emission from chloroplasts has been compared to the effect on subchloroplast particles. Non-cyclic electron flow of both chloroplasts and subchloroplast particles was blocked with 3-(3,4-dichlorophenyl)-1,1-dimethylurea. 2,3,5,6-tetramethyl p-phenylenediamine-catalyzed cyclic electron flow increased the millisecond delayed emission by 2–4 times in both chloroplasts and subchloroplast particles. Uncoupling conditions which collapse only the pH gradient component of the proton motive force reduced the 2,3,5,6-tetramethyl p-phenylenediamine stimulation of delayed light in chloroplasts but not in particles. The 2,3,5,6-tetramethyl p-phenylenediamine stimulation of delayed light in particles was sensitive to uncoupling conditions which are presumed to destroy the transmembrane potential. Energy transfer inhibitors were without effect on the 2,3,5,6-tetramethyl p-phenylenediamine stimulation in both chloroplasts and particles. The 2,3,5,6-tetramethyl p-phenylenediamine stimulation of millisecond delayed emission appears to reflect the particular form of the proton motive force; in chloroplasts it seems to be correlated with the proton concentration gradient, whereas in particles it is more closely correlated with the transmembrane potential. 相似文献
18.
Parallel measurements of the rise in chlorophyll a fluorescence yield and delayed light emission decay, after a 10 ns saturating excitation flash, have been made in tris(hydroxymethyl)aminomethane-washed chloroplasts. Various electron donor systems (Mn 2+; ascorbate; reduced phenylenediamine and benzidine) were used in conjuction with different preillumination regimes to alter [ P+-680], the oxidized form of the Photosystem II reaction center chlorophyll a. Conditions giving rise to high [ P+-680] resulted in only a small rise in fluorescence yield, an inhibition of a 6 μs delayed light component, and an enhancement of a 60 μs component of delayed light emission. These results confirm the hypothesis that P+-680 acts as a quencher of fluorescence and that delayed light emission in the microsecond time range is due to the back reaction of P+-680 and Q ?. (Q is the first “stable” electron acceptor of Photosystem II.) Two preillumination flashes are required before the full effect of Tris washing is observed in the delayed light emission decay and fluorescence yield rise; this suggests that a capacity to hold two charges exists between the Tris block and P+-680. Tris washing has no direct effect on the movement of electrons from Z (the first electron donor to P+-680) to P+-680. Finally, Mn 2+ donates electrons to P+-680 via Z. 相似文献
19.
1. The inhibitory action of tetraphenylboron, a lipid-soluble anion, on the proton uptake, the photophosphorylation and the light-induced quenching of the fluorescence of 9-aminoacridine by spinach chloroplasts was studied.2. The inhibition of the three processes by tetraphenylboron was transient; the proton uptake was affected to a much smaller extent than either the photophosphorylation or the fluorescence quenching.3. The inhibitory effects of tetraphenylboron on the proton uptake and the fluorescence quenching of 9-aminoacridine were qualitatively the same in CF 1-depleted chloroplasts, that were recoupled with N, N′-dicyclohexylcarbodiimide (DCCD).4. The reversal of the fluorescence quenching of 9-aminoacridine upon addition of tetraphenylboron in the light was found to be very fast, being completed within the response time of the apparatus.5. The presence of tetraalkylammonium salts in the incubation medium prevented the inhibitory effect of tetraphenylboron.6. Tetraphenylboron disappeared from the chloroplast suspension in a light-dependent irreversible way; in the dark no ‘ptake’ of tetraphenylboron could be detected.7. The effects of tetraphenylboron may be explained by the presence of groups with a high affinity for tetraphenylboron in the membrane; these groups become protonated upon illumination of the chloroplasts. 相似文献
20.
The ultraweak light emission of isolated chloroplasts (Hidegand Inaba (1991) Photochem. Photobiol. 52: 137) was investigatedin comparison to delayed light emission. We compared the concentrationdependence and the spectral distribution of the light emittedfrom isolated chloroplasts stored in the dark for 10 s, 2 min(delayed light emission), 4 and 10 h (ultraweak light emission),respectively. In samples with low chlorophyll concentration, spectra of allemission phenomena were maximal at 685695 nm, but spectraof ultraweak light, especially that of long term (10 h) emission,were broader in the 700800 nm region than spectra ofdelayed light, indicating emission from a bigger variety ofchlorophyll molecules. The intensity of delayed light and short term (4 h) ultraweaklight exhibited a simple, saturating exponential dependenceon chlorophyll concentration, while long term (10 h) ultraweaklight emission was best described as a saturating exponentialcontaining a quadratic function of the concentration. This differencesuggests that long term ultraweak light emission is broughtabout by reactions distinct from the earlier described mechanismof electron transport related dark photoemission. (Received November 15, 1991; Accepted May 18, 1992) 相似文献
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