Short term effect of ultraviolet-B radiation on photosystem 2 photochemistry in the cyanobacteriumSynechococcus 6301

Exposure of intact cells of theSynechococcus 6301 to UV-B radiation induced a loss in photosystem 2 (PS 2) electron transport activity prior to the alteration in pigment complexes. Thus the degradation of PS 2 was not directly related to pigment alteration. Supported by Young Scientist grant No. SRJSYIB-05192 from the Department of Science and Technology, Goverment of India to SDSM.     

Integrity and Activity of Photosystem 2 Complexes Isolated from the Thermophilic Cyanobacterium Synechococcus Elongatus Using Various Detergents

The efficiency in selective extraction of photosystem (PS) 2 oxygen evolving complexes was compared among seven detergents. These were applied to thylakoid membranes of the thermophilic cyanobacterium Synechococcus elongatus. Used were five non-ionic detergents with one ionic and one zwitterionic for comparison. To compare the suitability and efficiency of the detergents the following properties of the extracts were examined: maximum rate of oxygen evolution with various electron acceptors, the relative variable fluorescence (F_V/F_M), the contamination of the extract with photosystem (PS) 1, and the status of the electron acceptor side of PS2 reaction centre. None of the detergents yielded a highly selective extraction of the PS2 complexes (negligible contamination with PS1) which would simultaneously display a high photochemical activity and high structural intactness. Heptylthioglucoside and dodecylmaltoside yielded the nearest approximation to the optimum result. Kinetic fluorometry was applied here for the first time to characterize the functional and structural properties of PS2 particles from cyanobacteria. This revised version was published online in August 2006 with corrections to the Cover Date.     

A single step separation of PS 1, PS 2 and chlorophyll-antenna particles from spinach chloroplasts

After solubilization of photosynthetic membranes by digitonin, three main protein pigment complexes were isolated by electrophoresis with deoxycholate as detergent.The band with the slowest mobility, fraction 1, had PS 1 activity and was devoid of PS 2 activity. This fraction was four times enriched in P700 when compared with chloroplasts. Fraction 1 had little chl b, a long wavelength absorption maximum in the red, a maximum of low temperature emission fluorescence at 730nm, and a circular dichroism spectrum characteristic of PS 1 enriched fraction.Fraction 2 exhibited a PS 2 activity and no PS 1 activity. It was enriched five times in PS 2 reaction centre and had little chl b and carotenoids. The absorption maximum was at 674 nm and the low temperature fluorescence emission maximum was at 700 nm. Fraction 2 might be useful PS 2 enriched particle because of the great stability of this fraction with regard to photochemical activity and also rapidity and simplicity of its preparation.Fraction 3, which had the fastest migration, was devoid of photochemical activities; It was rich in chl b and had the fluorescence and the circular dichroism spectrum characteristic of an antenna complex.Abbreviations PS 1 (2) photosystem 1 (2) - chl chlorophyll - car carotenoid - Q primary plastoquinone electron acceptor - P700 primary electron donor of PS 1 - P680 primary electron donor of PS 2 - K₃Fe(CN)₆ potassium ferricyanide - DCMU dichlorophenyldimethylurea - DCPIP dichlorophenolindophenol - DPC diphenyl-carbazide     

Effect of Iron Deficiency Induced Changes on Photosynthetic Pigments,Ribulose-1,5-Bisphosphate Carboxylase,and Photosystem Activities in Field Grown Grapevine (Vitis Vinifera L. cv. Pinot Noir) Leaves

The effect of iron deficiency on photosynthetic pigments, ribulose-1,5-bisphosphate carboxylase (RuBPC), and photosystem activities were investigated in field grown grapevine (Vitis vinifera L. cv. Pinot noir) leaves. The contents of chlorophyll (Chl) (a+b) and carotenoids per unit fresh mass showed a progressive decrease upon increase in iron deficiency. Similar results were also observed in content of total soluble proteins and RuBPC activity. The marked loss of large (55 kDa) and small (15 kDa) subunits of RuBPC was also observed in severely chlorotic leaves. However, when various photosynthetic electron transport activities were analysed in isolated thylakoids, a major decrease in the rate of whole chain (H₂O methyl viologen) electron transport was observed in iron deficient leaves. Such reduction was mainly due to the loss of photosystem 2 (PS2) activity. The same results were obtained when F_v/F_m was evaluated by Chl fluorescence measurements in leaves. Smaller inhibition of photosystem 1 (PS1) activity was also observed in both mild and severely chlorotic leaves. The artificial electron donors, diphenyl carbazide and NH₂OH, markedly restored the loss of PS2 activity in severely chlorotic leaves. The marked loss of PS2 activity was evidently due to the loss of 33, 23, 28-25, and 17 kDa polypeptides in iron deficient leaves.     

Chromium increases photosystem 2 activity in <Emphasis Type="Italic">Brassica juncea</Emphasis>

In 7-d-old seedlings of Brassica juncea chromium (VI) promoted photosystem 2 (PS 2) mediated photoreactions. The increase in PS 2 activity in the thylakoids from Cr-treated seedlings, in the presence of uncoupler (5 mM NH₄Cl), was similar to that recorded with the control thylakoids. Thus Cr enhanced PS 2 activity was not due to uncoupling of electron transport from photophosphorylation. Photon saturation kinetics revealed that the PS 2 activity of thylakoids from Cr-treated seedlings was significantly higher at almost all irradiances in comparison to that of controls. PS 2 activity of thylakoids from Cr-treated plants at 25 % of the saturating irradiance was in par with the PS 2 activity of the thylakoids from control plants at saturating irradiance. Thylakoids from both control and Cr-treated seedlings exhibited maximum PS 2 activity at pH 7.5. The PS 2 activity of thylakoids from Cr-treated plants remained high even at pH 8.0 and 8.5, demonstrating Cr enhances tolerance of PS 2 to alkaline pH.     

Fluoride affects distribution of absorbed excitation energy more in favour of photosystem 1

The effects of fluoride on the photosynthetic electron transport chain have been studied in spinach thylakoid membranes. Inhibition in photosystem (PS) 2 electron transport rates and a subsequent increase in PS 1 electron transport rate indicated a possibility of state transitions being a mechanism of fluoride action. This hypothesis was further confirmed by the increase in fluorescence emission F_735/685 at 77 K, a decrease in variable to maximum fluorescence ratio (F_v/F_m) at room temperature and increase in the absorption cross section of PS 1 suggesting that fluoride affects distribution of the excitation energy in favour of PS 1 at the expense of PS 2.     

Calcium and magnesium effect on divalent cation deficientHydrilla verticillata thylakoid electron transport activity

The role of divalent cations like magnesium (Mg²⁺) and calcium (Ca²⁺) was irrvestigated on energy distribution process ofHydrilla verticillata thylakoids. Effect of these cations was tested on relative quantum yield of photosystem (PS) II catalyzed electron transport activity, room and liquid nitrogen temperature fluorescence emission properties and thylakoid light scattering characteristics. The electron transport activity was found to be stimulated in the presence of these cations in a light intensity independent manner. The concentration of cation required for maximum stimulation was nearly 10–12 mM. Comparatively, Ca²⁺ was more effective than Mg²⁺. Cation induced stimulation in electron transport activity was not accompanied by increase in chlorophylla fluorescence intensity either at room (25°C) or liquid nitrogen (77°K) temperatures. Furthermore, 540 nm absorption and 90° light scattering properties of thylakoids remained insensitive towards divalent cations. These facts together suggest that divalent cations inHydrilla thylakoids are not effective in supporting the excitation distribution between the interacting photosystem complexes.     

Modification of excitation energy distribution to photosystem I by protein phosphorylation and cation depletion during thylakoid biogenesis in wheat

The effects of protein phosphorylation and cation depletion on the electron transport rate and fluorescence emission characteristics of photosystem I at two stages of chloroplast development in light-grown wheat leaves are examined. The light-harvesting chlorophyll a/b protein complex associated with photosystem I (LHC I) was absent from the thylakoids at the early stage of development, but that associated with photosystem II (LHC II) was present. Protein phosphorylation produced an increase in the light-limited rate of photosystem I electron transport at the early stage of development when chlorophyll b was preferentially excited, indicating that LHC I is not required for transfer of excitation energy from phosphorylated LHC II to the core complex of photosystem I. However, no enhancement of photosystem I fluorescence at 77 K was observed at this stage of development, demonstrating that a strict relationship between excitation energy density in photosystem I pigment matrices and the long-wavelength fluorescence emission from photosystem I at 77 K does not exist. Depletion of Mg²⁺ from the thylakoids produced a stimulation of photosystem I electron transport at both stages of development, but a large enhancement of the photosystem I fluorescence emission was observed only in the thylakoids containing LHC I. It is suggested that the enhancement of PS I electron transport by Mg²⁺-depletion and phosphorylation of LHC II is associated with an enhancement of fluorescence at 77 K from LHC I and not from the core complex of PS I.     

Glycinebetaine Stabilizes Photosystem 1 and Photosystem 2 Electron Transport in Spinach Thylakoid Membranes Against Heat Inactivation

Glycinebetaine, a compatible osmolyte of halotolerant plants and bacteria, partially protected photosystem (PS) 1 and PS2 electron transport reactions against thermal inactivation but with different efficiencies. In its presence, the temperature for half-maximal inactivation (t_1/2) was generally shifted downward by 3–12 °C. Glycinebetaine stabilized photoinduced oxygen evolving reactions of PS2 by protecting the tetranuclear Mn cluster and the extrinsic proteins of this complex. A weaker, although noticeable, stabilizing effect was observed in photoinduced PS2 electron transport reactions that did not originate in the oxygen-evolving complex (OEC). This weaker protection by glycinebetaine was probably exerted on the PS2 reaction centre. Glycinebetaine protected also photoinduced electron transport across PS1 against thermal inactivation. The protective effect was exerted on plastocyanin, the mobile protein in the lumen that carries electrons from the integral cytochrome b ₆ f complex to the PS1 complex. This revised version was published online in June 2006 with corrections to the Cover Date.     

Effects of high temperature and low pH on photosystem 2 photochemistry in spinach thylakoid membranes

The effects of temperature (25–45 °C) and pH (7.5–5.5) on photosystem (PS) 2 was studied in spinach (Spinacia oleracea L.) thylakoid membranes using chlorophyll a fluorescence induction kinetics. In high temperature and low pH treated thylakoid membranes a decline in the variable to maximum fluorescence ratio (F_v/F_m) and PS 2 electron transport rate were observed. More stacking in thylakoid membranes, studied by digitonin fractionation method, was observed at low pH, while the degree of unstacking increased under high temperature conditions. We conclude that the change in pH does not significantly affect the donor/acceptor side of PS 2 while high temperature does. Fluorescence emission spectra at 77 K indicated that low pH is associated with energy redistribution between the two photosystems while high temperature induced changes do not involve energy re-distribution. We suggest that both, high temperature and low pH, show an inhibitory effect on PS 2 but their mechanisms of action are different.     

Temperature acclimation of photosynthesis and related changes in photosystem II electron transport in winter wheat

Winter wheat (Triticum aestivum L. cv Norin No. 61) was grown at 25 degrees C until the third leaves reached about 10 cm in length and then at 15 degrees C, 25 degrees C, or 35 degrees C until full development of the third leaves (about 1 week at 25 degrees C, but 2-3 weeks at 15 degrees C or 35 degrees C). In the leaves developed at 15 degrees C, 25 degrees C, and 35 degrees C, the optimum temperature for CO(2)-saturated photosynthesis was 15 degrees C to 20 degrees C, 25 degrees C to 30 degrees C, and 35 degrees C, respectively. The photosystem II (PS II) electron transport, determined either polarographically with isolated thylakoids or by measuring the modulated chlorophyll a fluorescence in leaves, also showed the maximum rate near the temperature at which the leaves had developed. Maximum rates of CO(2)-saturated photosynthesis and PS II electron transport determined at respective optimum temperatures were the highest in the leaves developed at 25 degrees C and lowest in the leaves developed at 35 degrees C. So were the levels of chlorophyll, photosystem I and PS II, whereas the level of Rubisco decreased with increasing temperature at which the leaves had developed. Kinetic analyses of chlorophyll a fluorescence changes and P700 reduction showed that the temperature dependence of electron transport at the plastoquinone and water-oxidation sites was modulated by the temperature at which the leaves had developed. These results indicate that the major factor that contributes to thermal acclimation of photosynthesis in winter wheat is the plastic response of PS II electron transport to environmental temperature.     

Effects of Cucumber Mosaic Virus Infection on Photosynthetic Activities of Tobacco Leaves and Chloroplasts

Changes in photosynthetic activities were studied with tobacco (Nicotiana tabacum L.) leaves and chloroplasts infected by cucumber mosaic virus (CMV) at the top, middle and bottom located leaves. Net photosynthetic rate was reduced at all three positioned leaves, with the maximum reduction occurring at the top leaves (31.9% of control). The infected chloroplasts showed a reduction in electron transport rates of the whole chain electron transport, photosystem Ⅱ (PSⅡ) and photosystem Ⅰ (PSⅠ). Since the decline in the whole chain electron transport (15.6% of control, H2O→MV) closely paralleled the decline in PSⅡ activity (20.9% of control, H2O→PBQ), the inhibition of the latter was probably responsible for the overall decrease. Chlorophyll a fluorescence measurements showed a variable reduced fluorescence yield (Fv/Fo) which indicated that PSⅡ was impaired and the CO2 assimilation was disturbed by CMV infection. Fluorescence emission spectra at 77 K indicated that energy distribution between PSⅡ and PSⅠ was affected. F686/F734 of infected leaves and chloroplasts increased and the greatest increase (331.1% of control ) was found in the top leaves. These data may conclude that the infection inhibited mainly the PSⅡ activity.     

Light-dependent, chilling effects on phosphorylation of thylakoid proteins and consequences for associated photochemical activities in maize

When maize ( Zea mays L. cv. LG11) leaves are exposed to low temperatures and high light modifications to both photosystem 2 (PS2) and the light-harvesting chlorophyll a/b protein complex associated with photosystem 2 (LHC2) occur. This study examines the consequences of these modifications for phosphorylation of LHC2 and PS2 polypeptides and the associated changes in electron transport. Maize leaves were chilled at 5°C for 6 h under photon flux densities of 1 500 and 250 μmol m^-2 s^-1. Thylakoids were then isolated from the leaves and their abilities to phosphorylate LHC2 and PS2 polypeptides and modify electron transport activities were determined. Measurements of chlorophyll fluorescence induction in the thylakoids were also made. Thylakoids isolated from leaves chilled under high light and from leaves kept in the ambient growth environment had similar phosphoprotein profiles. However, polypeptide phosphorylation in thylakoids from the chilled leaves did not produce a decrease in PS2 electron transport. Chilling leaves under low light produced a decrease in the ability of isolated thylakoids to phosphorylate PS2, but not LHC2, polypeptides, which was not associated with any change in the phosphorylation-induced decrease in PS2 electron transport. Chilling under high, but not low, light appears to produce changes in membrane organisation that do not affect the ability of the thylakoids to phosphorylate PS2 and LHC2 polypeptides, but which do prevent the phosphorylation-induced decrease in excitation energy transfer from LHC2 to PS2.     

Role of Light-harvesting Complex 2 Dissociation in Protecting the Photosystem 2 Reaction Centres Against Photodamage in Soybean Leaves and Thylakoids

The protective role of light-harvesting complex 2 (LHC2) dissociation from photosystem 2 (PS2) complex was explored by the 5-p-fluorosulfonylbenzoyl adenosine (FSBA, an inhibitor of protein kinase) treatment at saturating irradiance (SI) in soybean leaves and thylakoids. The dissociation of some LHC2s from PS2 complex occurred after SI treatment, but FSBA treatment inhibited the dissociation as demonstrated by analysis of sucrose density gradient centrifugation of thylakoid preparation and low-temperature (77 K) chlorophyll (Chl) fluorescence. A significant increase in F₀ and decrease in F_v/F_m occurred after SI, and the two parameters could largely recover to the levels of dark-adapted leaves after subsequent 3 h in the dark, but they could not recover in the FSBA-treated leaves at SI. Neither the electron transport activity of PS2 nor the D1 protein amount in vivo had significant change after SI without FSBA, whereas FSBA treatment at SI could result in significant decreases in both the PS2 electron transport activity and the D1 protein amount. When thylakoids instead of leaves were used, the PS2 electron transport activity and the D1 protein amount declined more after SI with FSBA than without FSBA. The phosphorylation level of PS2 core proteins increased, while the phosphorylation level of LHC2 proteins was reduced after SI. Also, the phosphorylation of PS2 core proteins could be greatly inhibited by the FSBA treatment at SI. Hence in soybean leaf the LHC2 dissociation is an effective strategy protecting PS2 reaction centres against over-excitation and photodamage by reducing the amount of photons transferred to the centres under SI, and the phosphorylation of PS2 core proteins plays an important role in the dissociation.     

Stress-induced degradation of D1 protein and its photoprotection by DCPIP in isolated thylakoid membranes of barley leaf

Effects of various stress treatments such as NaCl, hydrogen peroxide, hydroxyl free radical, and high irradiance (HI, 1 000 μmol m⁻² s⁻¹) on the photosystem (PS) 2 mediated electron transport rate and the degradation of D1 protein in the thylakoid membranes of barley were studied. The applied stresses caused significant reduction in the PS 2-mediated electron transport and a degradation of D1 protein that was highest during the HI-treatment. Presence of 2,6-dichlorophenol indophenol (DCPIP), which is an artificial electron acceptor from water, significantly minimizes the HI-induced deleterious effect on the PS 2-mediated electron transport rate, disarrangement of PS machinery, and degradation of the D1 protein. HI in the absence of an acceptor resulted in production of reactive oxygen species due to electron transfer to oxygen.     

Senescence Retarding Effect of Metal Ions: Pigment and Protein Contents and Photochemical Activities of Detached Primary Leaves of Wheat

Al³⁺ significantly delayed the loss of chlorophyll (Chl), protein, and carotenoids when compared to K⁺ and Mg²⁺ during dark-induced senescence of detached primary leaves of Triticum aestivum. Thylakoid membranes isolated from Al³⁺ - treated leaves showed a better retention of photosystem (PS) 2, PS1, and whole chain electron transport activities than thylakoids of K⁺- or Mg²⁺-treated leaves. These ions protected the electron transport activities and restored the DCMU-dependent fluorescence increase of thylakoid membranes in a valency-dependent manner. Al³⁺ also delayed the change of excitation energy distribution during senescence.     

The Chlorophyll-Protein Complexes Resolved from Ultrasonically Broken Thylakoid Memb-ranes of Blue-Green Algae

When the thylakoid membranes of blue-green algae were broken by ultrasonic vibrations and subjected to polyacrylamide gel electrophoresis at 4℃, six green zones were resolved. They were designated as CPIa, CPlb, CPI; CPal, CPa2, and FC. The absorption spectrum of CPI had a red maximum at 674 nm and a peak in the blue at 435 nm. It was identified as PS chlorophyll a-protein Complex, but was contaminated with minor PSⅡ which was implied by the appearance of fluorescence emission peak at 680 nm besides the main one at 725 nm at 77 K. The spectral properties of CPIa and CPlb were similar to that of CPl. The absorption spectra of CPa1 and CPa2 were similar, both having red maxima at 667 nm and peaks in the blue at 431.5 nm. Their fluorescence emission had the same peaks at 684 nm at 77 K indicating that they belonged to PSⅡ. It was recognized that CPal of 47 kD is the reaction center complex of photosystem Ⅱ and CPa2 of 40 kD is the internal antenna complex of photosystem Ⅱ. The spectral characteristics of the chlorophyll-protein complexes resolved by ultrasonic method were similar to those of the same complexes resolved by SDS solubilization, except the absorbance positions of CPa1 and CPa2 in the blue peak and the red one which shifted to blue about 3–5 nm. It was calculated that in thylakoid membranes of blue-green algae 40.93% chlorophyll was in PSⅠ, while 38.78% of chlorophyll in PSⅡ. The difference of chlorophyll contents between PSⅠ and PSⅡ was only 2.15%. Concerning the fact that minor PSⅡ compound remained in the part of PSⅠ zones, it might be concluded that the distribution of chlorophyll between PSⅠ and PSⅡ in blue-green algae was equal. This result was in agreement with the hypothesis that PSⅠ and PSⅡ operates in series in photosynthetic electron transport.     

Light-stress-related changes in the properties of photosystem I

Light-stress-related changes in photosystem I (PS I) were analyzed in photoautotrophic cultured cells of Marchantia polymorpha L. High light treatment (30␣h; 1300 μmol photons · m⁻² · s⁻¹) reduced the PS I-mediated electron-transport rate by more than 50% and the photochemical efficiency of photosystem II (PS II) by about 35%. In photoinhibited cells, 76% of the PS II centers remained closed in low light, which is in agreement with a preferential impairment of PS I. Our data indicate that excessive linear electron transport is a cause of the loss in PS I activity. Two PS I forms could be isolated by sucrose-gradient ultracentrifugation of mildly solubilized thylakoid membranes. After high-light treatment one of these forms, which showed a larger light-harvesting complex (LHC) I antenna and a specific association of LHC IIb, was enriched. The effect could be suppressed by blockage of linear electron transport. It is suggested that PS I inactivation and state transitions caused the change in PS I organisation. Received: 19 August 1996 / Accepted: 24 October 1996     

Photosynthetic Electron Transport,Photophosphorylation, and Antioxidants in Two Ecotypes of Reed (Phragmites Communis Trin.) from Different Habitats

We compared chloroplast photochemical properties and activities of some chloroplast-localised enzymes in two ecotypes of Phragmites communis, swamp reed (SR, C₃-like) and dune reed (DR, C₄-like) plants growing in the desert region of north-west China. Electron transport rates of whole electron transport chain and photosystem (PS) 2 were remarkably lower in DR chloroplasts. However, the electron transport rate for PS1 in DR chloroplasts was more than 90 % of the activity similar in the SR chloroplasts. Activities of Mg²⁺-ATPase and cyclic and non-cyclic photophosphorylations were higher in DR chloroplasts than in the SR ones. The activities of chloroplast superoxide dismutase (SOD) and ascorbate peroxidase (APX), both localised at or near the PS1 complex and serving to scavenge active oxygen around PS1, and the content of ascorbic acid, a special substrate of APX in chloroplast, were all higher in DR chloroplasts. Hence reed, a hydrophytic plant, when subjected to intense selection pressure in dune habitat, elevates its cyclic electron flow around PS1. In consequence, it provides extra ATP required by C₄ photosynthesis. Combined high activities of active oxygen scavenging components in DR chloroplasts might improve protection of photosynthetic apparatus, especially PS1, from the damage of reactive oxygen species. This offers new explanation of photosynthetic performance of plant adaptation to long-term natural drought habitat, which is different from those, subjected to the short-term stress treatment or even to the artificial field drought.     

Effects of NaCl stress on photochemical activity and thylakoid membrane polypeptide composition of a salt-tolerant and a salt-sensitive rice cultivar

NaCl stress (200 mM) inhibited the electron transport activity of photosystem 2 (PS2) more than that of PS1. The degree of electron transport activity inhibition was lower in the salt-tolerant cultivar Pokkali than in the salt-sensitive cultivar Peta. The polypeptide composition of the thylakoid membrane and PS2 particles did not change after NaCl treatment but there was a difference in polypeptide compositions of thylakoid membrane and PS2 particles between the two cultivars. PS2 particles of cv. Pokkali contained more 33-kDa and 43-kDa polypeptides than cv. Peta. Additionally, PS2 particles after NaCl treatment showed deficiency of 23-kDa outside polypeptides of PS2.