Selective inhibition of photosystem II in spinach by tobacco mosaic virus: an effect of the viral coat protein

Leaves of Spinacia oleracea inoculated with tobacco mosaic virus (TMV) strain PV230 develop mild chlorotic and mosaic symptoms of infection. Thylakoid membranes isolated from these infected leaves showed a reduced Fv/Fm ratio for chlorophyll fluorescence kinetics, at 25 degrees C. The photosystem II (PS II)-mediated electron-transport rate was inhibited 50%, whereas PS I activity was unaffected by virus infection. Protein analysis indicated that TMV coat protein was associated with thylakoids, in particular with the PS II fraction. The results demonstrate that TMV-infected S. oleracea shows inhibition of photosynthetic electron transport through PS II. We propose that the inhibition of photosynthetic activity results from the association of viral coat protein with the PS II complex.     

Novel Evidence for a Reversible Dissociation of Light-harvesting Complex Ⅱ from Photosystem Ⅱ Reaction Center Complex Induced by Saturating Light Illumination in Soybean Leaves

After saturating light illumination for 3 h the potential photochemical efficiency of photosystem Ⅱ (PSⅡ) (Fv/Fm, the ratio of variable to maximal fluorescence) decreased markedly and recovered basically to the level before saturating light illumination after dark recovery for 3 h in both soybean and wheat leaves, indicating that the decline in Fv/Fm is a reversible down-regulation. Also, the saturating light illumination led to significant decreases in the low temperature (77 K) chlorophyll fluorescence parameters F685 (chlorophyll a fluorescence peaked at 685 nm ) and F685/F735 (F735, chlorophyll a fluorescence peaked at 735 nm) in soybean leaves but not in wheat leaves. Moreover,trypsin (a protease) treatment resulted in a remarkable decrease in the amounts of PsbS protein (a nuclear gene psbS-encoded 22 kDa protein) in the thylakoids from saturating light-illuminated (SI), but not in those from darkadapted (DT) and dark-recovered (DRT) soybean leaves. However, the treatment did not cause such a decrease in amounts of the PsbS protein in the thylakoids from saturating light-illuminated wheat leaves. These results support the conclusion that saturating light illumination induces a reversible dissociation of some light-harvesting complex Ⅱ (LHCⅡ) from PSⅡ reaction center complex in soybean leaf but not in wheat leaf.     

Effect of methylmercury on primary photosynthesis processes in green microalgae Chlamydomonas reinhardtii

The sensitivity of green microalgae Chlamydomonas reinhardtii to methylmercury chloride (MeHg) and chloride mercury (HgCl2) was evaluated by measuring chlorophyll fluorescence parameters by the pulse-amplitude-modulation (PAM) fluorometry. It was shown that MeHg at concentrations above 1 microM decreased the Fv/Fm ratio, which characterizes the maximal efficiency of energy utilization in photosystem II. The degree of inhibition depended on the time of treatment and was always higher under illumination conditions (50 microE.m-2.s-1) than under dark conditions. A similar regularity was observed for the delta F/Fm' ratio, which characterizes the real efficiency of energy storage at the given intensity of the photosynthesis-exciting light. Incubation with 5 microM HgCl2 for 5 h did not affect both ratios. The decrease in Fm at constant F0 as well as changes in the fast fluorescence kinetics after MeHg treatment of algae cells indicated the damage on the donor side of photosystem II and the damage of the electron transfer from QA to QB. The reduction of photochemical fluorescence quenching (qN) under MeHg treatment is also evidence of the increase in the fraction of closed reaction centers (QA-). At the same time, increase in the steady-state level of P700 photooxidation indicated a disturbance of electron transfer between photosystems. The present study demonstrates that methylmercury treatment damaged the photosynthetic electron transfer chain at several sites. The inhibitory effect of methylmercury is much stronger than the effect of mercury chloride on photosynthetic processes.     

花后不同时期弱光和高温胁迫对小麦旗叶荧光特性及籽粒灌浆进程的影响

在田间试验条件下研究了花后不同时期弱光和高温胁迫对小麦旗叶荧光特性及籽粒灌浆进程的影响.结果表明,弱光处理3 d后,旗叶PSⅡ最大光化学效率（Fv/Fm）和光合速率（Pn）显著降低,但PSⅡ实际光化学效率（ΦPSⅡ）、荧光光化学猝灭系数（qP）和非光化学猝灭（NPQ）与对照相比差异较小;高温处理3 d后,Fv/Fm、Pn、ΦPSⅡ和qP均极显著降低, NPQ升高幅度较小.胁迫解除后,灌浆前期（花后8～10 d）弱光和高温处理后的旗叶荧光参数和光合速率略有恢复,但灌浆中期（花后15～17 d）处理后的各参数始终呈下降趋势, 说明前期处理效应是可逆的,而中期处理加速其衰老进程.用Logistic方程对各处理的籽粒灌浆过程模拟明,弱光和高温处理后籽粒粒重的降低主要是平均灌浆速率、最大灌浆速率和渐增期灌浆速率显著降低所致.灌浆持续期、最大灌浆速率出现时间、缓增持续期和缓增期灌浆速率受弱光和高温影响较小.     

Effects of Strong Light and Active Oxygen on Photosynthesis in Soybean

With the use of chlorophyll fluorescence technique, it was found that the net photosynthetic oxygen evolution rate decreased after strong light (2 000 μmol· m-2·2-1 ) treatment for two hours in soybean ( Glycine max L. ) leaves. The chlorophyll fluorescence parameters, Fm/Fo, Fv/Fm, ФPSII, qp and qN decreased along with the increase of light intensity. In strong light, exogenous active oxygen H202、·OH and 'O2 were harmful to soybean leaves. The destruction of 'O2 and·OH to leaves was most evident, as was shown that Fv/Fm and PS H decreased significantly. The antioxidants DABCO, mannitol, ascorbate and histidine protected the leaves, but weakly, from strong light. In darkness, the SOD inhibitor sodium diethyldithiocar- bamate (DDC) had no significant effect on Fm/Fo and Fv/Fm, but NAN,, the ascorbate peroxidase (APX)inhibitor, significantly decreased Fm/Fo, Fv/Fm and ФPS II. In strong light, however, beth DDC and NaN3 reduced the above-mentioned fluorescence parameters, but NaN3 was more effective than DDC. The results suggested that photoinhibition did take place in soybean leaves under strong light, and it was related to active oxygen in vivo.     

草莓叶片光合作用对强光的响应及其机理研究

用便携式调制叶绿素荧光仪和光合仪研究了强光下草莓叶片荧光参数及表观量子效率的变化.结果表明,Fm、Fv/Fm、PSⅡ无活性反应中心数量和Q_A的还原速率在强光下降低,在暗恢复时升高;而PSⅡ反应中心非还原性Q_B的比例在强光下增加,在暗恢复时降低.上述荧光参数的变化幅度均以强光胁迫或暗恢复的前10 min最大.强光下Φ_PSII、ETR和qP先升高后降低,但qN先大幅度降低,然后小幅回升.强光处理4 h后,丰香和宝交早生的表观量子效率(AQY)分别降低了20.9%和37.5%;qE(能量依赖的非光化学猝灭)为NPQ(非光化学猝灭)的最主要成分.强光胁迫下丰香的Fo、Fm、Fv/Fm、Φ_PSII、ETR和AQY的变化幅度均明显比宝交早生小.DTT处理后,草莓叶片的Fm和Fv/Fm明显降低,Fo显著升高.可以认为,依赖叶黄素循环和类囊体膜质子梯度两种非辐射能量耗散在草莓叶片防御光损伤方面起着重要作用,丰香的光合机构比宝交早生更耐强光.     

水淹对水芹叶片结构和光系统Ⅱ光抑制的影响

通过探讨在水淹条件下水芹（Oenanthe javanica）叶片结构的变化以及出水对其光系统II功能和光抑制的影响,阐明水芹光合机构在水淹条件下及出水后死亡的可能原因。结果表明：水淹条件下新生沉水功能叶光系统Ⅱ（PSⅡ）最大光化学效率（Fv/Fm）、电子传递活性与对照叶片差异很小,但水淹使气生功能叶的Fv/Fm显著降低;植株总生物量呈负增长趋势;活体弱光条件下,沉水叶出水后2小时叶片相对含水量（RWC）和Fv/Fm无显著变化;中等光强和强光条件下其RWC和Fv/Fm迅速降低;离体条件下,5小时的中等光强对沉水叶的Fv/Fm影响不显著,在随后的弱光下能恢复到出水时的初始状态;强光能使沉水叶的Fv/Fm大幅降低,且弱光下不能恢复到出水时的初始水平;在解剖结构上,水芹沉水叶的叶片总厚度、上下表皮厚度和气孔大小都显著低于气生叶,而且沉水叶没有明显的栅栏组织分化,但是沉水叶上表皮的气孔密度显著高于气生叶。研究结果表明,水淹使水芹原气生叶PSⅡ功能迅速衰退,但对新生沉水叶片影响很小。水芹植株出水后,沉水叶片结构变化使其在光下保水能力下降,而强光导致了光合机构的光抑制和反应中心失活。田间条件下两者共同作用则加剧了对叶片光合机构的破坏,进而致使其死亡。     

温州蜜柑叶片光合作用光抑制的保护机理

晴天条件下,使用便携式调制荧光仪和分光光度计观察了温州蜜柑叶片光合作用光抑制发生过程中几个主要荧光参数（初始荧光F0、最大荧光Fm、PSⅡ的光化学效率Fv/Fm、非光化学猝灭qN及其快相qNf和慢相qNs）、电子传递速率（ETR）和玉米黄素相对含量的日变化,结果表明,随着光强的增强,ETR、qN及其qNr与qNs以及玉米黄素相对含量升高,Fv/Fm、Fm和F0下降。用DTT处理后,qNs较对照明显下降,F0较对照明显上升,可以认为,柑橘在光合作用日变化中存在依赖于叶黄素循环和类囊体膜质子梯度两种非辐射能量耗散方式,而且它们在防御光破坏方面起着重要作用。     

Temperature, evapotranspiration and primary photochemical responses of apple leaves to hail

The objective of this work was to examine immediate physiological plant responses to hail and subsequent recovery in terms of evapotranspiration, leaf temperature and primary photochemical processes using apple as a model crop. Thermal emission pictures were taken in darkness to avoid interference from stomatal movements; temperature gradients were identified in concentric rings around sites of hail injury, with a distinct drop in temperature of up to 2.3 degrees C in the center immediately after the induced hail injury. This was due to enhanced evapotranspiration from the injured tissue. Six to twelve minutes after hail injury, the initial decrease in leaf temperature partially reversed. Chlorophyll fluorescence kinetics of light-adapted leaves showed a dramatic decrease in effective photosynthetic electron transport rate (ETR), from 20.5 to 9.0 micromol electron m(-2)s(-1) within 5 min from hail injury, and a rapid recovery to 14.1 micromol electron m(-2)s(-1) within the next 5 min. After 7h, ETR partially recovered to 17.4 micromol electron m(-2)s(-1). An initial drop in non-photochemical efficiency (NPQ) from 1.07 to 0.90 units within 5 min after hail injury was followed by a sharp increase to 1.67 units after another 5 min. During the next hour, NPQ gradually decreased to the initial level. This indicates increased thermal dissipation in photosystem II (PS II) as a protective mechanism against incident excessive energy in the leaves with closed stomata for 1h after hail injury. In contrast to the fluorescence kinetics of light-adapted leaves, maximum quantum yield Fv/Fm of PSII in the dark-adapted state remained unchanged at 0.79-0.81 relative units for the first 5 min after hail injury. Thereafter, Fv/Fm slowly declined to 0.75 within 1h, and to a trough of 0.73 at 3h. Seven hours after hail injury, Fv/Fm values were at 0.76, indicating partial recovery of PS II efficiency. The discrepancy in the dynamics of ETR and Fv/Fm responses may be explained by the formation of alternative electron sinks such as reactive oxygen species, particularly superoxides, which withdraw electrons from the photosynthetic transport, resulting in apparently higher values of calculated ETR.     

Preparation and functional characterization of thylakoids from Arabidopsis thaliana

A protocol for the isolation of functional thylakoids from Arabidopsis thaliana leaves was developed. The critical factor in obtaining active, coupled and stable preparation is the inclusion of EDTA and EGTA in the grinding buffer. Preparations were characterized with respect to the whole or partial electron transport chain, ATP/NADPH, ATP/O₂ and PS II/chlorophyll ratios. Sensitivity to a light-chill photoinhibitory treatment was also determined by evaluating the decrease in both maximal photochemical efficiency (Fv/Fm) and in electron transport rate. This revised version was published online in June 2006 with corrections to the Cover Date.     

Significance of molecular crowding in grana membranes of higher plants for light harvesting by photosystem II

Significance of molecular crowding in grana thylakoids of higher plants on photosystem II function was studied by 'titrating' the naturally high protein density by fusing unilamellar liposomes of the native lipid mixture with isolated grana membranes (BBY). The incorporation of lipids was monitored by equilibrium density gradient centrifugation and two-dimensional thin layer chromatography. The excitonic coupling between light-harvesting (LHC) II and photosystem (PS) II was analysed by chlorophyll a fluorescence spectroscopy. The fluorescence parameters Fv/Fm and Fo clearly depend on the protein density indicating the importance of molecular crowding for establishing an efficient excitonic protein network. In addition the strong dependency of Fo on the protein density reveals weak interactions between LHCII complexes which could be important for dynamic adjustment of the photosynthetic apparatus in higher plants.     

夜间低温胁迫对两种生长光强下藤黄幼苗叶片荧光特征和活性氧代谢的影响

对两种不同生长光强下（自然光的8％和50％）西双版纳热带雨林木本植物藤黄（Garcinia han-buryi）幼苗经夜间低温（4℃）处理后荧光特性和活性氧代谢的研究结果表明,低温使藤黄叶片光合机构PSⅡ原初光能转化效率（Fv/Fm）、PSⅡ非环式电子传递的量子效率（ФPSⅡ）、非光化学猝灭系数（NPQ）下降,原初荧光（F0）上升。低温胁迫消除后,生长在50％光强下藤黄叶片的Fv/Fm和F0在3d后仍不能完全恢复,而生长在8％光强下藤黄叶片的Fv/Fm和F0基本恢复,说明低温使生长在8％光强下藤黄的光合机构PSⅡ反应中心受到可逆失活,而生长在50％光强下藤黄的光合机构受到氧化伤害。随着低温胁迫时间的延长,两种生长光强藤黄叶片活性氧保护酶（SOD,CAT,APX）的活性虽升高,但O2^-的生成速率、H2O2和MDA含量积累增加。而在恢复阶段,生长在8％光强比生长在50％光强下藤黄叶片的活性氧含量下降得快,进一步说明生长在高光强的植物比生长在低光强的植物受低温伤害大。     

不同氮素水平下二氧化碳加富对草莓叶片光抑制的影响

用便携式调制叶绿素荧光仪和光合仪研究了强光下不同供氮水平（12、4和0.4 mmol·L^-1）和不同CO₂浓度下（700和390 μl·L^-1）丰香草莓叶片的荧光参数及净光合速率的变化.结果表明,CO₂和氮素对草莓叶片光抑制有明显的互作效应.在富CO₂下,12 mmol·L^-1供氮水平的草莓叶片净光合速率升高了62.7%,4和0.4 mmol·L^-1供氮水平则分别降低了7.4%和21.3%;12 mmol·L^-1供氮水平的F_m和F_v/F_m在强光胁迫时降辐减小,暗恢复时F_m和F_v/F_m恢复程度提高,而4和0.4 mmol·L^-1供氮水平却相反.表明氮素供应不足时草莓叶片在富CO₂环境下光合作用出现适应性下调,光抑制增强.     

Development of photosynthetic apparatus and respiration in pea seedlings during greening as influenced by toxic concentration of lead

Detached leaves of 14 day-old dark-grown pea seedlings were immersed with their cut ends either in water (control) or in 20 mM Pb(NO₃)₂ solution. They were exposed to continuous illumination during 24 and 48 h. The formation of PSII primary photochemistry in thylakoids was determined in vivo by measuring changes in values of parameters of chlorophyll a fast fluorescence kinetics: Fo, Fm, Fv, Fv/Fm and t 1/2. The amount of lead accumulation in leaves, content of chlorophylls and carotenoids and rates of CO₂ uptake in light and evolution in darkness (Pn-net photosynthesis and DR - dark respiration respectively) were determined. It has been found that with the exception of Fo, values of Fv, Fm and Fv/Fm were reduced by Pb²⁺. The values of t 1/2 were significantly larger in Pb²⁺ treated leaves. Decrease in the chlorophyll a fluorescence parameters was paralleled with the strong inhibition by this metal the biosynthesis of chlorophyll a and b but less of the carotenoids. Pb²⁺ drastically reduced Pn but had a stimulatory action on DR after 24 h and small inhibition of DR after 48 h exposure of leaves to this metal. As a consequence, after 48 h of greening the ratio of DR/Pn of control leaves was 0.45 whereas in Pb²⁺ treated leaves 2.7. It is proposed that DR in leaves plays a protective role against damage of Pn by Pb²⁺. Protection can be due to the supply the respiratory derived reductant and ATP to carry out cell metabolism upon reduced photosynthesis.     

水淹对水芹叶片结构和光系统II光抑制的影响

通过探讨在水淹条件下水芹(Oenanthe javanica)叶片结构的变化以及出水对其光系统II功能和光抑制的影响, 阐明水芹光合机构在水淹条件下及出水后死亡的可能原因。结果表明: 水淹条件下新生沉水功能叶光系统II(PSII)最大光化学效率(Fv/Fm) 、电子传递活性与对照叶片差异很小, 但水淹使气生功能叶的Fv/Fm显著降低; 植株总生物量呈负增长趋势; 活体弱光条件下, 沉水叶出水后2小时叶片相对含水量(RWC)和Fv/Fm无显著变化; 中等光强和强光条件下其RWC和Fv/Fm迅速降低; 离体条件下, 5小时的中等光强对沉水叶的Fv/Fm影响不显著, 在随后的弱光下能恢复到出水时的初始状态; 强光能使沉水叶的Fv/Fm大幅降低, 且弱光下不能恢复到出水时的初始水平; 在解剖结构上, 水芹沉水叶的叶片总厚度、上下表皮厚度和气孔大小都显著低于气生叶, 而且沉水叶没有明显的栅栏组织分化, 但是沉水叶上表皮的气孔密度显著高于气生叶。研究结果表明, 水淹使水芹原气生叶PSII功能迅速衰退, 但对新生沉水叶片影响很小。水芹植株出水后, 沉水叶片结构变化使其在光下保水能力下降, 而强光导致了光合机构的光抑制和反应中心失活。田间条件下两者共同作用则加剧了对叶片光合机构的破坏, 进而致使其死亡。     

Photosynthetic Properties of Photosystem II in Arabidopsis thaliana lpa1 Mutant

In a previous study, we characterized a high chlorophyll fluorescence lpa1 mutant of Arabidopsis thaliana, in which approximately 20% photosystem (PS) II protein is accumulated. In the present study, analysis of fluorescence decay kinetics and thermoluminescence profiles demonstrated that the electron transfer reaction on either the donor or acceptor side of PSII remained largely unaffected in the lpa1 mutant. In the mutant, maximal photochemical efficiency (Fv/Fm, where Fm is the maximum fluorescence yield and Fv is variable fluorescence) decreased with increasing light intensity and remained almost unchanged in wild-type plants under different light conditions. The Fv/Fm values also increased when mutant plants were transferred from standard growth light to low light conditions. Analysis of PSII protein accumulation further confirmed that the amount of PSII reaction center protein is correlated with changes in Fv/Fm in lpa1 plants. Thus, the assembled PSII in the mutant was functional and also showed increased photosensitivity compared with wild-type plants.(Author for correspondence. Tel: +86 (0)10 6283 6256; Fax: +86 (0)10 8259 9384; E-mail: zhanglixin@ibcas.ac.cn)     

Light inactivation of functional photosystem II in leaves of peas grown in moderate light depends on photon exposure

To determine the dependence of in vivo photosystem (PS) II function on photon exposure and to assign the relative importance of some photoprotective strategies of PSII against excess light, the maximal photochemical efficiency of PSII (Fv/Fm) and the content of functional PSII complexes (measured by repetitive flash yield of oxygen evolution) were determined in leaves of pea (Pisum satlvum L.) grown in moderate light. The modulation of PSII functionality in vivo was induced by varying either the duration (from 0 to 3 h) of light treatment (fixed at 1200 or 1800 mol photons · m^-2 · s^-1) or irradiance (from 0 to 3000 mol photons · m^-2 · s^-1) at a fixed duration (1 h) after infiltration of leaves with water (control), lincomycin (an inhibitor of chloroplast-encoded protein synthesis), nigericin (an uncoupler), or dithiothreitol (an inhibitor of the xanthophyll cycle) through the cut petioles of leaves of 22 to 24-day-old plants. We observed a reciprocity of irradiance and duration of illumination for PSII function, demonstrating that inactivation of functional PSII depends on the total number of photons absorbed, not on the rate of photon absorption. The Fv/Fm ratios from photoinhibitory light-treated leaves, with or without inhibitors, declined pseudo-linearly with photon exposure. The number of functional PSII complexes declined multiphasically with increasing photon exposure, in the following decreasing order of inhibitor effect: lincomycin > nigericin > DTT, indicating the central role of D1 protein turnover. While functional PSII and Fv/Fm ratio showed a linear relationship under high photon exposure conditions, in inhibitor-treated leaves the Fv/Fm ratio failed to reveal the loss of up to 25% of the total functional PSII under low photon exposure. The loss of this 25% of less-stable functional PSII was accompanied by a decrease of excitation-energy trapping capacity at the reaction centre of PSII (revealed by the fluorescence parameter, 1/Fo-1/Fm, where Fo and Fm stand for chlorophyll fluorescence when PSII reaction centres are open and closed, respectively), but not by a loss of excitation energy at the antenna (revealed by the fluorescence parameter, 1/Fm). We conclude that (i) PSII is an intrinsic photon counter under photoinhibitory conditions, (ii) PSII functionality is mainly regulated by D1 protein turnover, and to a lesser extent, by events mediated via the transthylakoid pH gradient, and (iii) peas exhibit PSII heterogeneity in terms of functional stability during photon exposure.Abbreviations D1 protein psbA gene product - DTT dithiothreitol - Fo chlorophyll fluorescence corresponding to open PSII reaction centres - Fv, Fm variable and maximum fluorescence after dark incubation, respectively - Fs, Fm steady-state and maximum fluorescence during illumination, respectively - P680 reactioncentre chlorophyll and primary electron donor of PSII - PS photosystem Financial support of this work by Department of Employment, Education and Training/Australian Research Council International Research Fellowships Program (Korea) is gratefully acknowledged.     

高温胁迫对柑橘光合速率和光系统Ⅱ活性的影响

用红外CO2分析仪和叶绿素荧光仪测定了温州蜜柑和脐橙叶片的净光合速率(Pn)、初始荧光(Fo)、最大光能转换效率(Fv／Fm)及电子传递速率(ETR)．结果表明,与常温(25℃)相比,高温胁迫(38～40℃)使温州蜜柑和脐橙叶片的Pn、Fv／Fm及ETR下降,Fo升高．胁迫25d后温州蜜柑和脐橙叶片的Pn分别下降55．6％和39．8％．Fv／Fm下降22．0％和6．7％,ETR下降55．0％和41．5％,Fo分别上升了113．8％和14．9％．柑橘经高温胁迫后,在25℃下处理10d,叶片的Pn、Fv／Fm、Fo及ETR恢复明显．这些结果说明柑橘的光合速率下降与PSⅡ反应中心失活有关．     

Performance index as an indicator of the physiological state of trees in urban forest ecosystems

Based on the measurements of fluorescence of bark chloroplasts by means of PAM and PEA fluorometers, the information capacity of the methods for assessing the physiological state of Tilia cordata L. from the maximum quantum efficiency of PS II photochemistry (Fv/Fm) and the performance index (PI) has been compared. The measurements were performed on annual shoots of linden trees growing in different environment. It was shown that the chlorophyll content in the bark of shoots growing near the busy urban street was twice less compared with trees growing out of the city. On the trees from the unsafe environment, a small decrease in the relative fluorescence variable (Fv/Fm) was registered, and there was a significant statistical deviation of this value compared to control trees. It was found that the PI and its constituent parameters calculated on the basis of light fluorescence induction curve (PEA-method) are more informative and allow one to recognize changes in the primary energy transformation processes in PS II when they are comparatively small. The results of our work show that PI can be used as a sensitive and a rapid test to evaluate the physiological state of trees and other plant objects even under minor environmental changes.     

Photoinhibition and recovery of photosynthesis in leaves of Vitis berlandieri and Vitis rupestris

Photoinhibition of photosynthesis was studied in Vitis berlandieri and Vitis rupestris leaves under controlled conditions (irradiation of detached leaves to about 1900 micromol m(-2) s(-1)). The degree of photoinhibition was determined by means of the ratio of variable to maximum chlorophyll (Chl) fluorescence (Fv/Fm) and electron transport measurements. The potential efficiency of PS2, Fv/Fm declined, Fo increased significantly in leaves of V. berlandieri, while Fo decreased in V. rupestris. In isolated thylakoids, the rate of whole chain and PS2 activity markedly decreased in high light irradiated more in leaves of V. berlandieri than in leaves of V. rupestris. A smaller inhibition of PS1 activity was also observed in both leaves. In the subsequent dark incubation, fast recovery was observed in both leaves and reached maximum PS2 efficiencies similar to those observed in non-photoinhibited leaves. The artificial exogenous electron donors DPC, NH2OH and Mn2+ failed to restore the high light induced loss of PS2 activity in V. berlandieri leaves, while DPC and NH2OH significantly restored in V. rupestris leaves. It is concluded that high light inactivates on the donor side of PS2 and acceptor side of PS2 in V. rupestris and V. berlandieri leaves, respectively. Quantification of the PS2 reaction center protein D1 and 33 kDa protein of water splitting complex following high light exposure of leaves showed pronounced differences between V. berlandieri and V. rupestris leaves. The marked loss of PS2 activity in high light irradiated leaves was due to the marked loss of D1 protein and 33 kDa protein in V. berlandieri and V. rupestris leaves, respectively.