Chlorophyll formation and the development of photosynthesis in illuminated etiolated pea leaves

Summary The protein synthesis inhibitors chloramphenicol and terramycin, and light of low intensity were used to retard the rate of chlorophyll formation in illuminated dark grown pea leaves. In the control leaves the onset of photosynthesis, as measured by carbon dioxide exchange of the whole leaves, and reduction of ferricyanide and metmyoglobin and photo-oxidation of ascorbate in isolated chloroplasts, was observed after 2–4 hours illumination. The photosynthetic activity of the treated leaves did not commence until 10–12 hours illumination had elapsed. In both the control and treated leaves the onset of photosynthesis occurred when the total chlorophyll content was 0.04 mg/g fresh weight. The precise point of photosynthetic inception was apparently more related to the attainment of a specific total chlorophyl content than to the ratio of chlorophyll a to chlorophyll b. A marked increase in the evolution of carbon dioxide in the light was observed in the treated leaves during the first 10 hours of greening. This observation could not be ascribed to photorespiration since the leaves did not possess an active photosystem. It is suggested that the enhanced respiration may have been due to the light-induced activation of synthetic pathways responsible for the formation of chloroplast constituents.The following abbreviations are used CMU 3(3-chlorophenyl)-1, 1-dimethylurea - DCIP dichlorophenol indophenol - PMS phenazine methosulphate - TRIS 2-amino-2-hydroxymethyl propane-1, 3-diol This work was supported by a Science Research Council studentship granted to R. J. Dowdell and submitted for the degree of Ph. D. of Bath University of Technology.     

Photophosphorylation in isolated maize bundle sheath chloroplasts and cells

Isolated maize bundle sheath chloroplasts showed substantial rates of noncyclic photophosphorylation. A typical rate of phosphorylation coupled to whole-chain electron transport (methylviologen or ferricyanide as acceptor) was 60 μmol per hour per milligram chlorophyll) with a coupling efficiency (P/e₂) of 0.6. Partial electron transport reactions driven by photosystem I or II supported phosphorylation with P/e₂ values of 0.2 to 0.3. Thus, two sites of phosphorylation seem to be associated with the photosynthetic chain in much the same way as in spinach chloroplasts.     

Influence of ascorbate and the Mehler peroxidase reaction on non-photochemical quenching of chlorophyll fluorescence in maize mesophyll chloroplasts

 Non-photochemical quenching of chlorophyll fluorescence (NPQ) and quantum yield of photosystem II (PSII) were studied with intact mesophyll chloroplasts of maize (Zea mays L.) during the initial minutes of illumination using the pulse-modulated chlorophyll fluorescence technique. Non-photochemical quenching was rapidly reversible in the dark at any point during illumination, which is indicative of energy-dependent dissipation of energy (mediated via thylakoid ΔpH changes and ascorbate-dependent synthesis of zeaxanthin). In chloroplasts suspensions including 15 mM ascorbate in the medium, with addition of oxaloacetate and pyruvate, the PSII yield, rate of reduction of oxaloacetate and phosphorylation of pyruvate reached a maximum after approximately 2 min of illumination. Under these conditions, which promote phosphorylation and a decreased ΔpH across the thylakoid membrane, NPQ rose to a maximum after 2–3 min of illumination, dropped to a minimum after about 6 min, and then increased to a steady-state level. A rather similar pattern was observed when leaves were illuminated following a 30-min dark period. Providing chloroplasts with higher levels of ascorbate (60 mM), prevented the transient drop in NPQ. Anaerobic conditions or addition of potassium cyanide caused a decrease in PSII yield, providing evidence for operation of the ascorbate-dependent Mehler-peroxidase reaction. These conditions also strongly suppressed the transient drop in NPQ. Dithiothreitol, an inhibitor of violaxanthin de-epoxidase, caused a large drop in NPQ even in the presence of high levels of ascorbate. The results suggest that the decline of NPQ occurs in response to an increase in lumen pH after initiation of phosphorylation, that this decline can be suppressed by conditions where ascorbate is not limiting for violaxanthin de-epoxidase, and that the increase of NPQ after such a decline is the result of development of energy dissipation in PSII reaction centers. Received: 13 August 1999 / Accepted: 17 September 1999     

Changes in the Ability of Photophosphorylation and Activities of Surface-Bound Adenosine Triphosphatase and Ribulose Diphosphate Carboxylase of Chloroplasts Isolated from the Barley Leaves Senescing in Darkness

Dark-induced aging of detached primary leaves of 11-day-old barley seedlings brings about a significant decline in the rates of ferricyanide [Fe(CN)₆]^3? reduction and photophosphorylations of isolated chloroplasts. Ferricyanide-supported noncyclic photophosphorylation is somewhat more susceptible to leaf aging than phenazine methosulfate (PMS)-supported cyclic phosphorylation. Non-latent membrane-bound adenosine triphosphatase (ATPase) and ribulosediphosphate carboxylase (RuDPCase) lose about half of their initial activities after 24 h, while during this period the electron transport and photophosphorylation activities are much less affected. Also, the loss of RuDPCase is almost complete, while chloroplasts still exhibit a significant level of [Fe(CN)₆]^3? reduction and photophosphorylations after 7 days of dark incubation. This would suggest that the enzymatic dark reactions are more sensitive to aging stress than the primary photochemical reactions of chloroplasts. Studies on the effect of divalent cations such as Mg²⁺ and Ca²⁺ on non-latent ATPase activity revealed that the dark stressed aging of detached leaves brings about a time dependent alteration in the response of this enzyme to Mg²⁺, but not to Ca²⁺. The former showed inhibitory as well as stimulatory response, whereas the latter always caused the usual stimulation. Addition of kinetin (50 μM) ensured retention of [Fe(CN)₆]^3? reduction, photophosphorylations and RuDPCase activity in chloroplasts during leaf aging, but it failed to preserve the initial loss in the activity of the non-activated membrane-bound ATPase.     

Rapid light curves: A new fluorescence method to assess the state of the photosynthetic apparatus

Photosynthetic electron transport rates (ETR), calculated from chlorophyll fluorescence parameters, were compared in long term light and dark adapted as well as photoinhibited Pisum sativum leaves using a novel chlorophyll fluorescence method and a new instrument: rapid light curves (RLC) generated with the MINI-PAM. RLCs are plots of ETRs versus actinic irradiances applied for 10 s. Large changes in maximum electron transport rates (ETR_max) were observed when leaves were shifted from dark to moderate light, or from dark to photoinhibitory light and vice versa. Maximum ETRs were very low following long term dark adaptation, but increased to maximum levels within 8 to 15 minutes of illumination. It took more than 3 hours, however, to return irradiance-exposed leaves to the fully dark adapted state. Quenching analysis of RLCs revealed large q_E development in long-term dark adapted leaves accounting for the low ETRs. Leaves photoinhibited for 3 hours had similarly reduced ETRs. In these leaves, however, q_I was largely responsible for this reduction. Actinic irradiance exposures and saturating flashes affected leaves with different irradiance histories differently.     

Appearance of Photochemical Activity in Isolated Chloroplasts from Far-red-illuminated Leaves of Phaseolus vulgaris

Chloroplast development was followed in intact bean leaves illuminatedwith far-red light by extracting chloroplasts at various timesto assay photosynthetic activities. Photochemical activity wasdetected in isolated chloroplasts prior to the times which werepreviously reported for intact leaf discs. Cyclic phosphorylationwas observed in isolated chloroplasts after 8 h of far-red illuminationwhile non-cyclic electron transport and phosphorylation weremeasurable after 12 and 16 h of illumination respectively. TheP/2e ratios were less than 0.5 after 24 h of far-red exposurebut approached a value of 1.0 by 60 h of illumination. Ammoniumchloride (10^–3 M) had little effect on electron transportin isolated chloroplasts until after 24 h of far-red illumination.Chlorophyll a accumulated slowly from the onset of far-red illuminationwhile chlorophyll b was not detected until after 48 h of far-redexposure. Leaf fresh weight increased four-fold over the 60h illumination period. Electron microscopy of isolated chloroplasts from far-red-illuminatedleaves indicated the presence of unfused primary thylakoidsby 12 h of exposure and prolamellar bodies throughout the entire60 h illumination period. Grana were not observed in isolatedchloroplasts nor were they induced by a 2 min exposure of thechloroplasts to 172 000 lx of white light. O₂ evolution in leaf discs of far-red-illuminated plants wasmeasurable after 16 h of illumination, attained a maximum valueby 36 h of far-red exposure, and then declined. Net CO₂ fixationwas observed in leaf discs after 8 h of far-red illuminationand the rates remained constant for an additional 16 h, beforeincreasing at least two-fold.     

Dependence of photosynthetic rates on leaf density thickness in deciduous woody plants grown in sun and shade

Comparisons of photosynthetic rates were made on leaves of ten species of woody dicotyledons grown in the field under full sun or under a canopy which transmitted approximately 18% of full light. Photosynthesis and dark respiration were measured and compared on various bases: area, chlorophyll, fresh weight of lamina, density thickness (fresh weight per unit area), and protein.

Light-saturated photosynthesis per unit area or unit chlorophyll was about 1.5 times greater in the sun leaves than in the shade leaves and essentially equal per unit fresh weight or unit protein. Sun leaves were thicker but the enzymes per unit fresh weight remained constant as thickness varied. Chlorophyll per unit area remained about constant; chlorophyll per unit fresh weight varied inversely with changes in leaf thickness. Thus, density thickness variation is important in photosynthetic adaptation to sun and shade. This is also shown by the relationship between light-saturated photosynthesis per unit area and density thickness.

     

Light scattering,chlorophyll fluorescence and state of the adenylate system in illuminated spinach leaves

Scattering of green light and chlorophyll fluorescence by spinach leaves kept in a stream of air or nitrogen were compared with leaf adenylate levels during illumination with blue, red or far-red light. Energy charge and ATP-ADP ratios exhibited considerable variability in different leaves both in the dark and in the light. Variability is explained by different possible states of the reaction oxidizing triose phosphate or reducing 3-phosphoglycerate. Except when oxygen levels were low, there was an inverse relationship between light scattering and chlorophyll fluorescence during illumination with blue or red light. When CO₂ was added to a stream of CO₂-free air, chlorophyll fluorescence increased, sometimes after a transient decrease, and both light scattering and leaf $\text{ATP}\text{ADP}$ ratios decreased. Similar observations were made when air was replaced by nitrogen under blue or high-intensity red light. Under these conditions, over-reduction caused inhibition of electron transport and phosphorylation in chloroplasts. However, when air was replaced by nitrogen during illumination with low-intensity red light or far-red light, light scattering increased instead of decreasing. Under these light conditions, $\text{ATP}\text{ADP}$ ratios were maintained in the light. They decreased drastically only after darkening. Although $\text{ATP}\text{ADP}$ ratios responded faster than light scattering or the slow secondary decline of chlorophyll fluorescence due to illumination, it appeared that in the steady state, light scattering and chlorophyll fluorescence are useful indicators of the phosphorylation state of the leaf adenylate system at least under aerobic conditions, when chloroplast and extrachloroplast adenylate systems can effectively communicate.     

Effects of sulfur on the photosynthesis of intact leaves and isolated chloroplasts of sugar beets

Effects of sulfur on photosynthesis in sugar beets (Beta vulgaris L. cv. F58-554H1) were studied by inducing sulfur deficiency and determining changes in the photosynthesis of whole attached leaves and of isolated chloroplasts. The rates of photosynthetic CO₂ uptake by intact leaves, photoreduction of ferricyanide, cyclic and noncyclic photophosphorylation of isolated chloroplasts, and the rate of CO₂ assimilation by ribulose diphosphate carboxylase, decreased with decrease in total leaf sulfur from 2500 to about 500 μg g⁻¹ dry weight. Sulfur deficiency reduced photosynthesis through an effect on chlorophyll content, which decreased linearly with leaf sulfur, and by decreasing the rate of photosynthesis per unit chlorophyll. There was only a small effect of sulfur deficiency on stomatal diffusion resistance to CO₂ until leaf sulfur decreased below 1000 μg g⁻¹ when stomatal resistance became a more significant proportion of the total diffusion resistance to CO₂. Light respiration rates were positively correlated with photosynthesis rates and dark respiration was unchanged as leaf sulfur concentrations declined.     

Photosynthetic apparatus in chilling-sensitive plants

The levels of both tightly and loosely bound Mn in chloroplasts from fresh, cold and dark stored as well as illuminated leaves of Lycopersicon esculentum Mill. were studied in relation to Hill reaction activity. The tightly bound Mn pool represents one third of the total Mn content in chloroplasts isolated from the fresh leaves, and its level does not change following cold storage and illumination of leaves. Upon cold and dark storage of leaves the loss from the chloroplasts of about 40%–50% of the total amount of Mn is accompanied by an almost complete inactivation of the Hill reactions, as studied with water as an electron donor, as well as by the appearance of an EPR signal characteristic of free Mn²⁺ ions. Following illumination of such leaves, the restroration of Hill reaction activity is accompanied by an increase in the total Mn content in chloroplasts of up to 70%–80% of the Mn level measured in the fresh leaves and by disappearance of the EPR signal. In contrast, aging in the cold of isolated chloroplasts does not affect their Mn content. The addition of manganese does not result in the restoration of Hill reaction activity in chloroplasts from cold stored leaves but causes a restoration of this activity inhibited by linolenic acid. The data suggest that the loosely bound Mn pool (extractable with Tris) can be differentiated into two fractions: (1) one functionally inactive in electron transport and (2) one responsible for restoration of Hill reaction activity. Mn of the latter fraction (about 45% of the total Mn content) probably originates from the free Mn ions present in the interior of the chloroplasts following the cold and dark storage of leaves and from Mn reincorporated into chloroplasts from the cytoplasm. Incorporation of Mn from both these sources into thylakoid membrane to form a functionally active, loosely bound Mn pool proceeds during the illumination of leaves and results in the restoration of Hill reaction activity inhibited following the storage of leaves in dark and cold.Abbreviations Chl chlorophyll - DCIP 2,6-dichlorophenolindophenol - Diquat 1,1-ethylene 2,2-dipiridylium dibromide - EPR electron paramagnetic resonance - FFA free fatty acid - MV methyl viologen, N,N-dimethyl-4,4 dipyridyldihydrochloride - Tris tris-(hydroxymethyl) aminomethane     

Photophosphorylation Associated with Photosystem II: II. Effects of Electron Donors, Catalyst Oxidation, and Electron Transport Inhibitors on Photosystem II Cyclic Photophosphorylation

Incubation of KCN-Hg-NH₂OH-inhibited spinach (Spinacia oleracea L.) chloroplasts with p-phenylenediamine for 10 minutes in the dark prior to illumination produced rates of photosystem II cyclic photophosphorylation up to 2-fold greater than the rates obtained without incubation. Partial oxidation of p-phenylenediaine with ferricyanide produced a similar stimulation of ATP synthesis; addition of dithiothreitol suppressed the stimulation observed with incubation. Addition of ferricyanide in amounts sufficient to oxidize completely p-phenylenediamine failed to inhibit completely photosystem II cyclic activity. This is due at least in part to the fact that the ferrocyanide produced by oxidation of p-phenylenediamine is itself a catalyst of photosystem II cyclic photophosphorylation. N,N,N′N′-Tetramethyl-p-phenylenediamine catalyzes photosystem II cyclic photophosphorylation at rates approaching those observed with p-phenylenediamine. The activities of both proton/electron and electron donor catalysts of the photosystem II cycle are inhibited by dibromothyoquinone and antimycin A. These findings are interpreted to indicate that photosystem II cyclic photophosphorylation requires the operation of endogenous membrane-bound electron carriers for optimal coupling of ATP synthesis to electron transport.     

Cyclic and Noncyclic Photophosphorylation in Isolated Guard Cell Chloroplasts from Vicia faba L

High rates of both cyclic and noncyclic photophosphorylation were measured in chloroplast lamellae isolated from purified guard cell protoplasts from Vicia faba L. Typical rates of light-dependent incorporation of ³²P into ATP were 100 and 190 micromoles ATP per milligram chlorophyll per hour for noncyclic (water to ferricyanide) and cyclic (phenazine methosulfate) photophosphorylation, respectively. These rates were 50 to 80% of those observed with mesophyll chloroplasts. Noncyclic photophosphorylation in guard cell chloroplasts was completely inhibited by 3-(3,4-dichlorophenyl)-1,1-dimethylurea supporting the notion that photophosphorylation is coupled to linear electron flow from photosystem II to photosystem I. Several lines of evidence indicated that contamination by mesophyll chloroplasts cannot account for the observed photophosphorylation rates.

A comparison of the photon fluence dependence of noncyclic photophosphorylation in mesophyll and guard cell chloroplasts showed significant differences between the two preparations, with half saturation at 0.04 and 0.08 millimole per square meter per second, respectively.

     

Effects of calcium on the photosynthesis of intact leaves and isolated chloroplasts of sugar beets

Effects of calcium on photosynthesis in sugar beets (Beta vulgaris L. cv. F58-554H1) were studied by inducing calcium deficiency and determining changes in CO₂ uptake by attached leaves, electron transport, and photophosphorylation by isolated chloroplasts, and CO₂ assimilation by ribulose diphosphate carboxylase extracts. Calcium deficiency had no significant effect on leaf CO₂ uptake, photoreduction of ferricyanide, cyclic or noncyclic ATP formation of isolated chloroplasts, or on ribulose diphosphate carboxylase CO₂ assimilation, when the rates were expressed per unit chlorophyll. When expressed per unit leaf area CO₂ uptake increased by about 15% in low calcium leaves. The most noticeable effect of calcium deficiency was reduction in leaf area: low calcium had no effect on dark respiratory CO₂ evolution, on leaf diffusion resistance, or on mesophyll resistance to CO₂. We concluded that only small amounts of calcium are required for normal photosynthetic activity of sugar beet leaves.     

Cyclic and noncyclic photophosphorylation during the ontogenesis of high-light and low-light leaves of Sinapis alba

Noncyclic electron transport to ferricyanide and photophosphorylation as well as the methylviologen mediated aerobic and anaerobic photophosphorylation with dichlorophenolindophenol-ascorbate as the electron donor of photosystem I were measured during the development of high-light and low-light adapted leaves of Sinapis alba. Anaerobic methylviologen-catalyzed phosphorylation is more than twice as high as aerobic phosphorylation. The difference between the rates of aerobic and anaerobic phosphorylation is sensitive to dibromothymoquinone. Thus, under anaerobic conditions, methylviologen mediates a cyclic phosphorylation including plastoquinone. All photochemical activities of high-light chloroplasts are about twice as high as that of low-light chloroplasts and show a permanent decline with increasing plant age. The lower activities of low-light chloroplasts correlate with a decrease of electron transport components, such as cytochrome f. This indicates that the number of electron transport chains is decreased under low-light conditions and more chlorophyll molecules interact with one electrontransport chain.Abbreviations Asc ascorbate - Chl chlorophyll a+b - DBMIB 2,5-dibromo-3-methyl-6-isopropyl-p-benzoquinone - DCMU 3-(dichlorophenyl)-1,1-dimethylurea - DCPIP dichlorophenolindophenol - HL high light - LL low light - MV methylviologen - PhAR photosynthetically active radiation - PS photosystem     

Studies on Greening of Etiolated Seedlings

Evidence is given that a selective light-pretreatment of the embryonic axis exerts a deep influence on the greening in primary leaves of 8-day-old etiolated bean seedlings (Phaseolus vulgaris cv. Limburg). After a subsequent dark incubation of sufficient length and a final exposure of the entire plants to continuous illumination the lag phase of chlorophyll synthesis is completely removed. In particular the highly meristematic hook tissue seems to be responsible for this light effect. Lengthening of the dark period following pre-irradiation increased the capability of chlorophyll production in the main white light period, reaching its maximum after about 12 hours of darkness. The period of dark incubation for elimination of the lag phase is considerably longer in plants with shielded leaves than the length of the lag phase in etiolated seedlings of the same age, exposed entirely to continuous light. This difference may be explained by the synergistic effect between leaves and embryonic axis. Evidence for this interorgan cooperation is given by experiments with a selective light-pretreatment of leaves and embryonic axis. After a 5 min pre-exposure to white light of whole plants the leaves of some of the plants were shielded and these plants received a further pre-illumination of 2 hours on their embryonic axis. In all the pre-irradiated, etiolated plants the lag phase of chlorophyll synthesis was eliminated during the main white light period, following a dark incubation of 2 hours. Additional and preferential light activation of the embryonic axis during the pretreatment had no significant effect on chlorophyll production during the white light illumination after a 2 hours dark incubation, but resulted in a lower yield of chlorophylls after 18 hours dark incubation compared to the white light controls, receiving no selective light-pretreatment on the embryonic axis. From our results we can decisively conclude that a simultaneous light-pretreatment of both, leaves and embryonic axis, is more effective and beneficial for building up a capacity of chlorophyll synthesis in the leaves than either a selective light-pretreatment of the embryonic axis alone or a simultaneous pre-illumination of leaves and embryonic axis, immediately followed by an additional preirradiation of the embryonic axis. Therefore, we think that several photoactive sites are involved in de-etiolation processes of intact, etiolated seedings. Light activation of the embryonic axis stimulates the development of this organ and contributes to the greening processes in the leaf. At the same time, by irradiating the leaf, light activates the photo-sensitive site in the leaf itself, which also develops a capacity for chlorophyll synthesis. Both photo-acts are cooperative, explaining the enhanced chlorophyll production. Additional pre-irradiation of the embryonic axis after a short illumination of whole plants favours its own development and reduces the synthetic capacity of the leaf. A prolonged far-red pretreatment induces qualitatively the same response as white light. We assume that these effects on lag phase removal and chlorophyll production, induced in etiolated, primary bean leaves by selective irradiation of the embryonic axis, is a phytochrome-mediated process. Our results indicate a transmission of light-induced stimuli from one organ to another.     

Inhibition of oxygen evolution in chloroplasts by ferricyanide

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

Photophosphorylation and Carbon Dioxide Fixation by Chloroplasts Isolated from Populus deltoides

A system has been developed for the isolation of photosynthetically active chloroplasts from leaves of Populus deltoides. A high proportion of the chloroplasts appeared intact. The maximum rates of different photosynthetic processes were as follows: CO₂ fixation 3.5 micromoles per milligram chlorophyll per hour, noncyclic ATP synthesis 10 micromoles per milligram chlorophyll per hour, and cyclic ATP synthesis 300 micromoles per milligram chlorophyll per hour.     

Acceleration of the Decay of Membrane Potential after Energization of Chloroplasts in Intact Zea mays Leaves

The relationship between dissipation of the flash-induced membranepotential across the thylakoid membrane and the high energystate was studied in Zea mays leaves. The dark decay of theflash-induced 515-nm absorbance change was accelerated by shortpreillumination of the leaf. No acceleration of the decay bypreillumination was observed when leaves were incubated in argonor CO² gas or treated with DCMU. These effects of preilluminationand incubation were reversible. The delayed fluorescence from chlorophyll a was reversibly decreasedby incubating leaves in argon or CO² gas, though the modes ofdepression were somewhat different from each other. In leavesincubated in argon or CO² gas, the phase of slow decrease ofthe intensity of prompt fluorescence during illumination reversiblydisappeared. The results suggested that the dissipation of membrane potentialgenerated by a flash was accelerated after the energizationof chloroplasts in leaves, probably by increased H^– permeabilityof the thylakoid membrane. O2 was important in maintaining (indarkness) and forming (under illumination) the high energy statein chloroplasts in intact leaves. (Received October 1, 1980; Accepted December 15, 1980)     

Evidence against proton gradient formation being the cause of chlorophyll fluorescence quenching by N-methylphenazonium methosulfate.

In strong illumination, 3-(3, 4-dichlorophenyl)-1,1-dimethylurea (DCMU)-poisoned chloroplasts exhibit a high yield of chlorophyll fluorescence while P-700 turnover, proton uptake, and phosphorylation are inhibited and a pH gradient is undectectable. When 10muM N-methylphenazonium methosulfate (PMS) is included, the fluorescence yield in light is substantially reduced, and when 100 muM ascorbate is also included, the yield is diminished approximately to the level in darkness. Only very slight increases in P-700 turnover and proton uptake (but no detectable pH gradient) accompany the fluorescence yield decline. When 10muM PMS and 15 mM ascorbate are added to poisoned chloroplasts (the oxygen concentration being greatly reduced), P-700 turnover, proton uptake, the pH gradient and phosphorylation all reach high levels. In this case, the yield of chlorophyll fluorescence is low and is the same in both light and dark. Further addition of an uncoupler eliminates proton uptake, the pH gradient and phosphorylation but does not significantly elevate the fluorescence yield. From these observations we suggest that, in DCMU-poisoned chloroplasts, the fluorescence quenching with PMS occurrs by a mechanism unrelated to the generation of a phosphyorylation potential. With chloroplasts unpoisoned by DCMU, PMS quenches fluorescence and considerably stimulates proton uptake, the pH gradient and phosphorylation. However, in this case, PMS serves to restore net electron transport.     

The Effect of Light and Hormones on Leaf Senescence

With wheat leaves as material, the changes of superoxide dismutase (SOD), lipid peroxi-dation and membrane permeability during leaf senescence in light or dark, and treated withphytohormones (KT or ABA) have been studied. The changes of chlorophyll content, lipidperoxidation and fine structure of spinach chloroplasts senescing in light or dark have alsobeen studied. When leaves senesce in light, the activity of SOD increased at first then decreased. The increase of SOD activity was able to result from the synthesis of new protein. Lightwas found to delay the leaf senescence obviously but also accelerate leaf senescence by causinglipid peroxidation when prolonged the illumination time. The delay or acceleration of leafsenescence by exogenous hormones were observed, it may be due to the control of lipid peroxi-dation by adjusting the activity of SOD. O2-participated the chlorophyll decomposition andlipid peroxidation during chloroplasts senesce in light. A favourable role of light in mainta-lng the fine structure of isolated chloroplasts was clear.