PSII photochemistry, thermal energy dissipation, and the xanthophyll cycle in Kalanchoë daigremontiana exposed to a combination of water stress and high light

Kalanchoë daigremontiana, a CAM plant grown in a greenhouse, was subjected to severe water stress. The changes in photosystem II (PSII) photochemistry were investigated in water‐stressed leaves. To separate water stress effects from photoinhibition, water stress was imposed at low irradiance (daily peak PFD 150 μmol m^?2 s^?1). There were no significant changes in the maximal efficiency of PSII photochemistry (F_v/F_m), the traditional fluorescence induction kinetics (OIP) and the polyphasic fluorescence induction kinetics (OJIP), suggesting that water stress had no direct effects on the primary PSII photochemistry in dark‐adapted leaves. However, PSII photochemistry in light‐adapted leaves was modified in water‐stressed plants. This was shown by the decrease in the actual PSII efficiency (Φ_PSII), the efficiency of excitation energy capture by open PSII centres (F_v′/F_m′), and photochemical quenching (q_P), as well as a significant increase in non‐photochemical quenching (NPQ) in particular at high PFDs. In addition, photoinhibition and the xanthophyll cycle were investigated in water‐stressed leaves when exposed to 50% full sunlight and full sunlight. At midday, water stress induced a substantial decrease in F_v/F_m which was reversible. Such a decrease was greater at higher irradiance. Similar results were observed in Φ_PSII, q_P, and F_v′/F_m′. On the other hand, water stress induced a significant increase in NPQ and the level of zeaxanthin via the de‐epoxidation of violaxanthin and their increases were greater at higher irradiance. The results suggest that water stress led to increased susceptibility to photoinhibition which was attributed to a photoprotective process but not to a photodamage process. Such a photoprotection was associated with the enhanced formation of zeaxanthin via de‐epoxidation of violaxanthin. The results also suggest that thermal dissipation of excess energy associated with the xanthophyll cycle may be an important adaptive mechanism to help protect the photosynthetic apparatus from photoinhibitory damage for CAM plants normally growing in arid and semi‐arid areas where they are subjected to a combination of water stress and high light.     

Photoinhibition and xanthophyll cycle activity in bayberry (Myrica rubra) leaves induced by high irradiance

The effect of high irradiance (HI, photosynthetically active photon flux density of 1 300 μmol m⁻² s⁻¹) on net photosynthetic rate (P _N), chlorophyll fluorescence parameters, and xanthophyll cycle components were studied in fruit tree bayberry leaves. HI induced the photoinhibition and inactivation of photosystem 2 (PS2) reaction centres (RCs), which was characterized by decreased P _N, maximum yield of fluorescence after dark adaptation (F_m), photochemical efficiency of PS2 (F_v/F_m) and quantum yield of PS2 (Φ_PS2), and increased reduction state of Q_A (1-q_P) and non-photochemical quenching (NPQ). Initial fluorescence (F₀) showed a decrease after the first 2 h, and subsequently increased from the third hour exposure to HI. Furthermore, a greater increase in the ratio (F_i-F₀)/(F_p-F₀) which is an expression of the proportion of the Q_B non-reducing PS2 centres, whereas a remarked decrease in the slope of F_i to F_p which represents the rate of Q_A reduction was observed in leaves after HI exposure. Additionally, HI caused an increase in the pool size of the xanthophyll cycle pigments and sustained elevated contents of zeaxanthin (Z), antheraxanthin (A), and de-epoxidation state (DES) at the end of the irradiation period. During HI, decreased F_m, F_v/F_m, Φ_PS2, NPQ, slope of F_i to F_p, V+A+Z, and DES, and increased F₀, 1-q_P, ratio (F_i-F₀)/(F_p-F₀), and V were observed in dithiothreitol (DTT)-fed leaves compared to control ones under the same conditions. Hence photoinhibition caused by HI in bayberry was probably attributed to inactivation of PS2 RCs, and photoprotection from photodamage were mainly related to the xanthophyll cycle-dependent heat dissipation in excess photons.     

Kinetics of prolonged photoinhibition revisited: photoinhibited Photosystem II centres do not protect the active ones against loss of oxygen evolution

Photoinhibition of Photosystem II (PSII) in lincomycin-treated leaves begins as a first-order reaction, but fluorescence measurements have suggested that after prolonged illumination, the number of active PSII centres stabilizes to 15–20% of control. The stabilization has been interpreted to indicate that photoinhibited PSII centres protect the remaining active centres against photoinhibition (Lee, Hong and Chow, Planta 212:332–342, 2001). In an attempt to study the mechanism of this protection, we measured the reaction kinetics of photoinhibition in lincomycin-treated pumpkin (Cucurbita pepo L.) and pepper (Capsicum annuum L.) leaves in vivo. The light-saturated rate of PSII oxygen evolution, assayed from thylakoids and isolated from the treated leaves, was used as a direct measure of the number of remaining active PSII centres, and the fluorescence parameters F _V/F _M and (F _V/F _M)/F ₀ (=1/F ₀ − 1/F _M) were measured for comparison. To our surprise, no stabilization of PSII activity was observed and photoinhibition followed first-order kinetics until PSII activity had virtually declined to zero. A series of in vitro experiments was carried out to see whether stabilization of PSII activity occurs if a particular combination of light intensity and wavelength range is applied, or if a specific PSII preparation is used as experimental material. The results of the in vitro experiments confirmed the in vivo result about persistent first-order kinetics. We conclude that photoinhibited PSII centres offer no measurable protection against photoinhibition.     

Arabidopsis plants lacking PsbS protein possess photoprotective energy dissipation

It is commonly accepted that the photosystem II subunit S protein, PsbS, is required for the dissipation of excess light energy in a process termed ‘non‐photochemical quenching’ (NPQ). This process prevents photo‐oxidative damage of photosystem II (PSII) thus avoiding photoinhibition which can decrease plant fitness and productivity. In this study Arabidopsis plants lacking PsbS (the npq4 mutant) were found to possess a competent mechanism of excess energy dissipation that protects against photoinhibitory damage. The process works on a slower timescale, taking about 1 h to reach the same level of NPQ achieved in the wild type in just a few minutes. The NPQ in npq4 was found to display very similar characteristics to the fast NPQ in the wild type. Firstly, it prevented the irreversible light‐induced closure of PSII reaction centres. Secondly, it was uncoupler‐sensitive, and thus triggered by the ΔpH across the thylakoid membrane. Thirdly, it was accompanied by significant quenching of the fluorescence under conditions when all PSII reaction centres were open (F_o state)_. Fourthly, it was accompanied by NPQ‐related absorption changes (ΔA535). Finally, it was modulated by the presence of the xanthophyll cycle carotenoid zeaxanthin. The existence of a mechanism of photoprotective energy dissipation in plants lacking PsbS suggests that this protein plays the role of a kinetic modulator of the energy dissipation process in the PSII light‐harvesting antenna, allowing plants to rapidly track fluctuations of light intensity in the environment, and is not the primary cause of NPQ or a direct carrier of the pigment acting as the non‐photochemical quencher.     

Mechanistic differences in photoinhibition of sun and shade plants

Leaf discs of the shade plant Tradescantia albiflora Kunth grown at 50 μmol · m^?2 · s^?1, and the facultative sun/shade plant Pisum sativum L. grown at 50 or 300 μmol · m^?2, s^?1, were photoinhibited for 4 h in 1700 μmol photons m^?2 · s^?1 at 22° C. The effects of photoinhibition on the following parameters were studied: i) photosystem II (PSII) function; ii) amount of D1 protein in the PSII reaction centre; iii) dependence of photoinhibition and its recovery on chloroplast-encoded protein synthesis; and, iv) the sensitivity of photosynthesis to photoinhibition in the presence or absence of the carotenoid zeaxanthin. We show that: i) despite different sensitivities to photoinhibition, photoinhibition in all three plants occurred at the reaction centre of PSII; ii) there was no correlation between the extent of photoinhibition and the degradation of the D1 protein; iii) the susceptibility to photoinhibition by blockage of chloroplas-tencoded protein synthesis was much less in shade plants than in plants acclimated to higher light; and iv) inhibition of zeaxanthin formation increased the sensitivity to photoinhibition in pea, but not in the shade plant Tradescantia. We suggest that there are mechanistic differences in photoinhibition of sun and shade plants. In sun plants, an active repair cycle of PSII replaces photoinhibited reaction centres with photochemically active ones, thereby conferring partial protection against photoinhibition. However, in shade plants, this repair cycle is less important for protection against photoinhibition; instead, photoinhibited PSII reaction centres may confer, as they accumulate, increased protection of the remaining connected, functional PSII centres by controlled, nonphotochemical dissipation of excess excitation energy.     

Role of the xanthophyll cycle in photoprotection elucidated by measurements of light-induced absorbance changes,fluorescence and photosynthesis in leaves of Hedera canariensis

The role of the xanthophyll cycle in regulating the energy flow to the PS II reaction centers and therefore in photoprotection was studied by measurements of light-induced absorbance changes, Chl fluorescence, and photosynthetic O₂ evolution in sun and shade leaves of Hedera canariensis. The light-induced absorbance change at 510 nm (A₅₁₀) was used for continuous monitoring of zeaxanthin formation by de-epoxidation of violaxanthin. Non-radiative energy dissipation (NRD) was estimated from non-photochemical fluorescence quenching (NPQ).High capacity for zeaxanthin formation in sun leaves was accompanied by large NRD in the pigment bed at high PFDs as indicated by a very strong NPQ both when all PS II centers are closed (F'_m) and when all centers are open (F'_o). Such F_o quenching, although present, was less pronounced in shade leaves which have a much smaller xanthophyll cycle pool.Dithiothreitol (DTT) provided through the cut petiole completely blocked zeaxanthin formation. DTT had no detectable effect on photosynthetic O₂ evolution or the photochemical yield of PS II in the short term but fully inhibited the quenching of F_o and 75% of the quenching of F_m, indicating that NRD in the antenna was largely blocked. This inhibition of quenching was accompanied by an increased closure of the PS II reaction centers.In the presence of DTT a photoinhibitory treatment at a PFD of 200 mol m^-2 s^-1, followed by a 45 min recovery period at a low PFD, caused a 35% decrease in the photon yield of O₂ evolution, compared to a decrease of less than 5% in the absence of DTT. The F_v/F_m ratio, measured in darkness showed a much greater decrease in the presence than in the absence of DTT. In the presence of DTT F_o rose by 15–20% whereas no change was detected in control leaves.The results support the conclusion that the xanthophyll cycle has a central role in regulating the energy flow to the PS II reaction centers and also provide direct evidence that zeaxanthin protects against photoinhibitory injury to the photosynthetic system.Abbreviations F, F_m, F_o, F_v Fluorescence yield at actual degree of PS II center closure, when all centers are closed, when all centers are open, variable fluorescence - NPQ non-photochemical fluorescence quenching - NRD non-radiative energy dissipation - PFD photon flux density - Q_A primary acceptor PS II     

On the significance of photoinhibition of photosynthesis in the field and its generality among species

Photoinhibition was examined in naturally exposed willow leaves in the field. In the afternoon on clear and warm days, the quantum yield of electron transport, derived from gas exchange data, was decreased by 28%. Besides this photoinhibition, decreases in the photosynthetic capacity and in the stomatal conductance were also observed. Of these three limitations of carbon assimilation, photoinhibition was the major one at limiting light only.To investigate the generality of photoinhibition, shade- and sun-acclimated leaves of fourteen different species were compared in a laboratory study. Photoinhibition was quantified by fluorescence measurements following exposure to moderate and high light for 30 min. The extent of photoinhibition was inversely related to the photochemical quenching, q_p, during exposure (the proportion of open PS II traps). This relationship was the same independent of the species, the light-acclimation state of the leaf and the light intensity. However, sun- and shade-acclimated leaves occupied opposite sides of the relationship: the sun-leaves showed lower photoinhibition and higher q_p. The sun-leaves were also more competent than shade-leaves by showing faster recovery from a given level of photoinhibition.Abbreviations F₀, F_V, F_M, F_S minimal, variable, maximal and steady-state fluorescence - q_N, q_i total and photoinhibitory non-photochemical quenching of variable fluorescence - q_p photochemical quenching     

Mathematical modelling of the light response curve of photoinhibition of Photosystem II

The light response curves of the acceptor and donor side mechanisms of photoinhibition of Photosystem II were calculated, using Arabidopsis as a model organism. Acceptor-side photoinhibition was modelled as double reduction of Q_A, noting that non-photochemical quenching has the same effect on the quantum yield of Q_A double reduction in closed PSII centres as it has on the quantum yield of electron transport in open centres. The light response curve of acceptor-side photoinhibition in Arabidopsis shows very low efficiency under low intensity light and a relatively constant quantum yield above light saturation of photosynthesis. To calculate the light response curve of donor-side photoinhibition, we built a model describing the concentration of the oxidized primary donor P₆₈₀⁺ during steady-state photosynthesis. The model is based on literature values of rate constants of electron transfer reactions of PSII, and it was fitted with fluorescence parameters measured in the steady state. The modelling analysis showed that the quantum yield of donor-side photoinhibition peaks under moderate light. The deviation of the acceptor and donor side mechanisms from the direct proportionality between photoinhibition and photon flux density suggests that these mechanisms cannot solely account for photoinhibition in vivo, but contribution of a reaction whose quantum yield is independent of light intensity is needed. Furthermore, a simple kinetic calculation suggests that the acceptor-side mechanism may not explain singlet oxygen production by photoinhibited leaves. The theoretical framework described here can be used to estimate the yields of different photoinhibition mechanisms under different wavelengths or light intensities.     

Photoinhibition and Recovery of Photosynthesis in Canker-susceptible and Resistant Needles of Cypress (Cupressus sempervirens L.)

The aim of the present investigation was to test the hypothesis that the cypress canker caused by a fungus (Seiridium cardinale) infection induced effects on photosynthesis which could be related to photoinhibition and the process of recovery in susceptible and resistant needles. Photoinhibition of photosynthesis and recovery was studied in canker‐infected susceptible and resistant needles of cypress (Cupressus sempervirens L.) under controlled conditions (irradiation of detached needles to approximately 1900 μmol/m²/s). The degree of photoinhibition was determined by means of the ratio of variable to maximum chlorophyll (Chl) fluorescence (F_v/F_m) and electron transport measurements. The potential efficiency of photosystem (PS) II, F_v/F_m declined, and F_o increased significantly in canker‐susceptible needles, while F_o did not change in resistant needles. In isolated thylakoids, high light (HL) decreased the rate of whole chain and PS II activity markedly more in susceptible than in resistant needles. A smaller reduction of PS I activity was noticed only in susceptible needles. Upon subsequent dark incubation, fast recovery was noticed in both needle types and reached maximum rates of PS II efficiency similar to those noticed in non‐photoinhibited needles. The artificial exogenous electron donors such as diphenyl carbazide (DPC), NH₂OH and Mn²⁺ failed to restore the HL induced loss of PS II activity in susceptible needles, while DPC and NH₂OH significantly restored it in resistant needles. The results suggest that HL inactivates the donor side of PS II in resistant and the acceptor side of PS II in susceptible needles. The results on the quantification of the PS II reaction centre protein D1 and 33 kDa protein of water‐splitting complex following HL exposure showed pronounced differences between susceptible and resistant needles. The marked loss of PS II activity in HL‐irradiated needles was due to the marked loss of D1 protein in susceptible and 33 kDa protein in resistant needles, respectively.     

Site-specific mutations in the D1 polypeptide affect the susceptibility of Synechocystis 6803 cells to photoinhibition

Photoinhibition of photosystem II in the cyanobacterium Synechocystis 6803 was followed after site-specific mutagenesis of the D1 polypeptide. Mutations were created in the stromal/cytosolic loop connecting helices D and E. Two mutations E243K and CA1, a deletion of the three glutamates 242–244 and a substitution Q241H, were made in the putative cleavage area of the D1 polypeptide. A third mutation E229D was made in the PEST-like sequence. Mutants and control cells were illuminated and F_V/F_M was recorded. Compared to the control, the mutants were less photoinhibited. Fluorescence relaxation after a single flash was delayed in CA1. Restoration of F_V/F_M after photoinhibition in the mutants was totally dependent on protein synthesis while control cells were able to recover partially also when protein synthesis was inhibited. In addition, the protein synthesis-dependent recovery of CA1 was slowed down. Our results indicate a correlation between the mutated amino acids and photoinhibition of photosystem II.     

Resistance to the herbicide paraquat and increased tolerance to photoinhibition are not correlated in several weed species

Photoinhibition was examined in paraquat-resistant and paraquat-susceptible biotypes of Hordeum glaucum Steud., Hordeum leporinum Link., Arctotheca calendula (L.) Levyns., and Conyza bonariensis (L.) Cronq. Plants were photoinhibited at low temperature, and the extent of photoinhibition determined by O₂ evolution and 77 K fluorescence. No difference in the degree of photoinhibition was detected between paraquat-resistant and paraquat-susceptible biotypes for any of the species examined. C. bonariensis plants were also photoinhibited by treatment without CO₂ at either 21% (volume/volume) O₂ or 4% (volume/volume) O₂, and again no difference was observed between the paraquat-resistant and paraquat-susceptible biotypes in reduction of the ratio of variable fluorescence to maximal fluorescence. This is in contrast to a recent report (MAK Jansen, Y Shaaltiel, D Kazzes, O Canaani, S Malkin, J Gressel, [1989] Plant Physiol 91: 1174-1178 in which it was claimed that a paraquat-resistant biotype of C. bonariensis was more tolerant of photoinhibition than a paraquat-susceptible biotype. We conclude that paraquat-resistant biotypes of these plant species are not more tolerant of photoinhibition when compared with the paraquat-susceptible biotypes.     

Gap size effects on photoinhibition in understorey saplings in tropical rainforest

The sudden increase in irradiance after canopy disturbance in primary forest together with the accompanying increase in leaf temperatures is known to cause photoinhibition in shade acclimated foliage of understorey plants. We hypothesized that there is species specific variation among understorey saplings in the magnitude of photoinhibition in response to gap creation, which is related to their requirement for overstorey disturbance. Eleven more or less circular gaps were created varying in size from 60 up to 1459 m². Photoinhibition was assessed by determining predawn and midday F_v/F_m using chlorophyll fluorescence at two occasions during the first 3 weeks after creation of the gaps. The light environment was assessed using hemispherical photography. Five species that occurred in sufficient numbers in the understorey after gap creation were measured. They all showed an increase of photoinhibition with increasing gap size. Variation in exposure to direct sunlight within gaps contributed also to variation in photoinhibition. Dynamic photoinhibition, the overnight increase in F_v/F_m, was about 20% of total photoinhibition as measured at midday. The species responded quantitatively different. Oxandra asbeckii was most sensitive as evident from a decrease of predawn F_v/F_m from 0.79 in the understorey of undisturbed forest to 0.70 in the smallest and further to 0.41 in the largest gaps. Catostemma fragrans, the least sensitive species showed hardly any photoinhibition in the smallest gaps and less in the largest ones, whereas Lecythis concertiflora, Licania heteromorpha, and Chlorocardium rodiei had intermediate responses. Species rank order in sensitivity to photoinhibition was maintained across the whole range of gap sizes. The relationship between sensitivity to photoinhibition and species-specific gap size preference for regeneration is discussed.     

Photoinhibition of photosynthesis represents a mechanism for the long-term regulation of photosystem II

The obligate shade plant, Tradescantia albiflora Kunth grown at 50 mol photons · m^–2 s^–1 and Pisum sativum L. acclimated to two photon fluence rates, 50 and 300 mol · m^–2 · s^–1, were exposed to photoinhibitory light conditions of 1700 mol · m^–2 · s^–1 for 4 h at 22° C. Photosynthesis was assayed by measurement of CO₂-saturated O₂ evolution, and photosystem II (PSII) was assayed using modulated chlorophyll fluorescence and flash-yield determinations of functional reaction centres. Tradescantia was most sensitive to photoinhibition, while pea grown at 300 mol · m^–2 · s^–1 was most resistant, with pea grown at 50 mol · m^–2 · s^–1 showing an intermediate sensitivity. A very good correlation was found between the decrease of functional PSII reaction centres and both the inhibition of photosynthesis and PSII photochemistry. Photoinhibition caused a decline in the maximum quantum yield for PSII electron transport as determined by the product of photochemical quenching (q_p) and the yield of open PSII reaction centres as given by the steady-state fluorescence ratio, F_vF_m, according to Genty et al. (1989, Biochim. Biophys. Acta 990, 81–92). The decrease in the quantum yield for PSII electron transport was fully accounted for by a decrease in F_vF_m, since q_p at a given photon fluence rate was similar for photoinhibited and noninhibited plants. Under lightsaturating conditions, the quantum yield of PSII electron transport was similar in photoinhibited and noninhibited plants. The data give support for the view that photoinhibition of the reaction centres of PSII represents a stable, long-term, down-regulation of photochemistry, which occurs in plants under sustained high-light conditions, and replaces part of the regulation usually exerted by the transthylakoid pH gradient. Furthermore, by investigating the susceptibility of differently lightacclimated sun and shade species to photoinhibition in relation to q_p, i.e. the fraction of open-to-closed PSII reaction centres, we also show that irrespective of light acclimation, plants become susceptible to photoinhibition when the majority of their PSII reaction centres are still open (i.e. primary quinone acceptor oxidized). Photoinhibition appears to be an unavoidable consequence of PSII function when light causes sustained closure of more than 40% of PSII reaction centres.Abbreviations F_o and F_o minimal fluorescence when all PSII reaction centres are open in darkness and steady-state light, respectively - F_m and F_m maximal fluorescence when all PSII reaction centres are closed in darkand light-acclimated leaves, respectively - F_v variable fluorescence - (F_m-F_o) under steady-state light con-ditions - F_s steady-state fluorescence in light - Q_A the primary,stable quinone acceptor of PSII - q_Ne non-photochemical quench-ing of fluorescence due to high energy state - (pH); q_Ni non-photochemical quenching of fluorescence due to photoinhibition - q_p photochemical quenching of fluorescence To whom correspondence should be addressedThis work was supported by the Swedish Natural Science Research Council (G.Ö.) and the award of a National Research Fellowship to J.M.A and W.S.C. We thank Dr. Paul Kriedemann, Division of Forestry and Forest Products, CSIRO, Canberra, Australia, for helpful discussions.     

Photoinhibition of photosynthesis in intact kiwifruit (Actinidia deliciosa) leaves: Effect of temperature

Photoinhibition of photosynthesis was induced in attached leaves of kiwifruit grown in natural light not exceeding a photon flux density (PFD) of 300 mol·m^-2·s^-1, by exposing them to a PFD of 1500 mol·m^-2·s^-1. The temperature was held constant, between 5 and 35° C, during the exposure to high light. The kinetics of photoinhibition were measured by chlorophyll fluorescence at 77K and the photon yield of photosynthetic O₂ evolution. Photoinhibition occurred at all temperatures but was greatest at low temperatures. Photoinhibition followed pseudo first-order kinetics, as determined by the variable fluorescence (F _v) and photon yield, with the long-term steady-state of photoinhibition strongly dependent on temperature wheareas the observed rate constant was only weakly temperature-dependent. Temperature had little effect on the decrease in the maximum fluorescence (F _m) but the increase in the instantaneous fluorescence (F _o) was significantly affected by low temperatures in particular. These changes in fluorescence indicate that kiwifruit leaves have some capacity to dissipate excessive excitation energy by increasing the rate constant for non-radiative (thermal) energy dissipation although temperature apparently had little effect on this. Direct photoinhibitory damage to the photosystem II reaction centres was evident by the increases in F _o and extreme, irreversible damage occurred at the lower temperatures. This indicates that kiwifruit leaves were most susceptible to photoinhibition at low temperatures because direct damage to the reaction centres was greatest at these temperatures. The results also imply that mechanisms to dissipate excess energy were inadequate to afford any protection from photoinhibition over a wide temperature range in these shade-grown leaves.Abbreviations and symbols fluorescence yield correction coefficient - F _o, F _m, F _v instantaneous, maximum, variable fluorescence - K _D, K _F, K _P, K _T rate constants for non-radiative energy dissipation, fluorescence, photochemistry, energy transfer to photosystem I - PFD photon flux density - PSI, II photosystem I, II - _i photon yield of photosynthesis (incident light)     

Pulse amplitude modulated fluorescence in the green macrophytes,Codium adherens,Enteromorpha muscoides,Ulva gigantea and Ulva rigida,from the Atlantic coast of Southern Spain

The effect of ambient and enhanced solar radiation on the photosynthetic apparatus in four marine green macroalgae on the Southern coast of Spain (Strait of Gibraltar) was investigated using pulse amplitude modulation (PAM) fluorescence. The dependence of the fluorescence parameters on the irradiance of the actinic light was determined for all four species. It showed that maximal fluorescence after light adaptation (F_m′), photochemical quenching (q_P) and the photosynthetic quantum yield decreased in Enteromorpha muscoides with irradiance while non-photochemical quenching (q_N) rose continuously. In Ulva rigida the photosynthetic quantum yield dropped at irradiances above 4 W m⁻² but q_P did not decrease with increasing light. q_N quenching rose sharply above 37 W m⁻², and maximal fluorescence dropped above 1 W m⁻². In Ulva gigantea the yield dropped to zero at irradiances of 37 W m⁻², as did q_P at 53 W m⁻². q_N started from an intermediate level and increased to a maximum at the highest irradiances. In Codium adherens, the yield and q_P behaved similarly as in U. rigida, while q_N rose at much lower irradiances. All investigated algae suffered from photoinhibition even at their natural sites of growth when the sun is at high angles. The hypothesis that algae with flat thalli suffer more than those with massive ones was confirmed. Photoinhibition was less pronounced in U. rigida and C. adherens than in the other two species. After 1 h of exposure to solar radiation at the surface, the photosynthetic quantum yield decreased substantially in the surface algae E. muscoides and U. rigida. In both macroalgae, recovery of the photosynthetic quantum yield was almost complete after 2–3 h in the shade. Two other green algae from shaded habitats (U. gigantea and C. adherens) did not show complete recovery of the yield from photoinhibition. This confirms the second hypothesis that sun-adapted algae recover faster from photoinhibition than those adapted to shaded sites.     

Heterogeneity of Chlorophyll Fluorescence over Thalli of Several Foliose Macrolichens Exposed to Adverse Environmental Factors: Interspecific Differences as Related to Thallus Hydration and High Irradiance

Spatial heterogeneity of chlorophyll (Chl) fluorescence over thalli of three foliose lichen species was studied using Chl fluorescence imaging (CFI) and slow Chl fluorescence kinetics supplemented with quenching analysis. CFI values indicated species-specific differences in location of the most physiologically active zones within fully hydrated thalli: marginal thallus parts (Hypogymnia physodes), central part and close-to-umbilicus spots (Lasallia pustulata), and irregulary-distributed zones within thallus (Umbilicaria hirsuta). During gradual desiccation of lichen thalli, decrease in Chl fluorescence parameters (F_O - minimum Chl fluorescence at point O, F_P - maximum Chl fluorescence at P point, ₂ - effective quantum yield of photochemical energy conversion in photosystem 2) was observed. Under severe desiccation (>85 % of water saturation deficit), substantial thalli parts lost their apparent physiological activity and the resting parts exhibited only a small Chl fluorescence. Distribution of these active patches was identical with the most active areas found under full hydration. Thus spatial heterogeneity of Chl fluorescence in foliose lichens may reflect location of growth zones (pseudomeristems) within thalli and adjacent newly produced biomass. When exposed to high irradiance, fully-hydrated thalli of L. pustulata and U. hirsuta showed either an increase or no change in F_O, and a decrease in F_P. Distribution of Chl fluorescence after the high irradiance treatment, however, remained the same as before the treatment. After 60 min of recovery in the dark, F_O and F_P did not recover to initial values, which may indicate that the lichen used underwent a photoinhibition. The CFI method is an effective tool in assessing spatial heterogeneity of physiological activity over lichen thalli exposed to a variety of environmental factors. It may be also used to select a representative area at a lichen thallus before application of single-spot fluorometric techniques in lichens.     

Gibberellic-acid-responsive protoplasts from mature aleurone of Himalaya barley

The role of oxygen in the photoinactivation of the photosynthetic apparatus of Spinacia oleracea L. was investigated. Moderate irradiation (1200 mol photons m^-2s^-1) of spinach leaves in an atmosphere of pure nitrogen caused strong inhibition of subsequently measured net CO₂ assimilation, whereas considerably less photoinhibition was observed in the presence of low partial pressures (10–20 mbar) of O₂. The decrease in activity caused by anaerobiosis in the light was not based on stomatal closure; the decline of assimilation represents a photoinhibition, as activity was not impaired by low irradiation (80 mol photos m^-2s^-1). In contrast, gassing with pure N₂ in the dark caused strong inhibition. Electron-transport rates and chlorophyll-fluorescence data of thylakoids isolated from photoinhibited leaves indicated damage to the electron-transport system, in particular to photosystem II reaction centers. In vitro, photoinhibition in isolated thylakoid membranes was also strongly promoted by anaerobiosis. Photoinhibition of electron-transport rates under anaerobic conditions was characterized by a pronounced increase in the initial fluorescence level, F₀, of chlorophyll-fluorescence induction, in contrast to photoinhibition under aerobic conditions. The results are discussed in terms of two mechanisms of photoinhibition, one that is suppressed and a second that is promoted by oxygen.Abbreviations Chl chlorophyll - DCMU 3-(3, 4-dichlorophenyl)-1,1-dimethylurea - PSI, II photosystem I, II     

Cold hardening reduces photoinhibition of Eucalypts nitens and E. pauciflora at frost temperatures

Photoinhibition of photosynthesis at low temperatures was investigated in two species of subalpine eucalypt, Eucalypts nitens (Deane and Maiden) Maiden and E. pauciflora Sieb. ex Spreng. Imposition of an artificial cold-hardening treatment increased the frost tolerance of leaf tissue and increased tolerance to excess light. Cold-hardened seedlings of both species had a higher photosynthetic capacity than non-hardened seedlings at 6 and 16°C and lower levels of non-photochemical quenching (NPQ) at 20 and 5°C. Furthermore, hardened seedlings had faster rates of NPQ development at 5 and −3.5°C. An increase in minimal fluorescence, which indicates slowly reversible photoinhibition, was evident in all seedlings at −1.5 and −3.5°C but was less pronounced in hardened seedlings, with a threefold faster rate of development of NPQ, at −3.5°C than non-hardened seedlings. Hardened seedlings also recovered faster from photoinhibition at −3.5°C. Thus cold hardening increased tolerance to high light in these species. Differences between E. nitens and E. pauciflora in their response to excess light were small and significant only at −3.5°C. Faster recovery from photoinhibition of E. pauciflora was consistent with its occurrence in colder habitats than E. nitens. Received: 27 April 1997 / Accepted: 9 September 1997     

Two components of onset and recovery during photoinhibition of Ulva rotundata

Short-term (up to 5 h) transfers of shade-adapted (100 mol · m^–2 · s^–1) clonal tissue of the marine macroalga Ulva rotundata Blid. (Chlorophyta) to higher irradiances (1700, 850, and 350 mol · m^–2 · s^–1) led to photoinhibition of room-temperature chlorophyll fluorescence and O₂ evolution. The ratio of variable to maximum (F_v/F_m) and variable (F_v) fluorescence, and quantum yield () declined with increasing irradiance and duration of exposure. This decline could be resolved into two components, consistent with the separation of photoinhibition into energy-dissipative processes (photoprotection) and damage to photosystem II (PSII) by excess excitation. The first component, a rapid decrease in F_v/F_m and in F_v, corresponds to an increase in initial (F_o) fluorescence and is highly sensitive to 1 mM chloramphenicol. This component is rapidly reversible under dim (40 mol · m^–2 · s^–1) light, but is less reversible with increasing duration of exposure, and may reflect damage to PSII. The second (after 1 h exposure) component, a slower decline in F_v/F_m and F_v with declining F_o, appears to be associated with the photoprotective interconversion of violaxanthin to zeaxanthin and is sensitive to dithiothreitol. The accumulation of zeaxanthin in U. rotundata is very slow, and may account for the predominance of increases in F_o at high irradiances.Abbreviations and Symbols CAP chloramphenicol - DTT dithiothreitol - F_o, F_m, F_v initial, maximum, and variable fluorescence - quantum yield - PFD photon flux density - PSII photosystem II To whom correspondence should be addressedWe are grateful to O. Björkman and S. Thayer, Carnegie Institution of Washington, Stanford, Cal., USA, for analysis of xanthophyll pigments reported here. This research was supported by National Science Foundation grant OCE-8812157 to C.B.O. and J.R. Support for G.L. was provided by a NSF-CNRS (Centre National de la Recherche Scientifique) exchange fellowship.     

Curling during desiccation protects the foliose lichen Lobaria pulmonaria against photoinhibition

This study aims to assess the photoprotective potential of desiccation-induced curling in the light-susceptible old forest lichen Lobaria pulmonaria by using chlorophyll fluorescence imaging. Naturally curled thalli showed less photoinhibition-induced limitations in primary processes of photosynthesis than artificially flattened specimens during exposures to 450 μmol m⁻² s⁻¹ in the laboratory after both 12- (medium dose treatment) and 62-h duration (high dose treatment). Thallus areas shaded by curled lobes during light exposure showed unchanged values of measured chlorophyll fluorescence parameters (F _V/F _M, Φ_{PS II}), whereas non-shaded parts of curled thalli, as well as the mean for the entire flattened thalli, showed photoinhibitory limitation after light treatments. Furthermore, the chlorophyll fluorescence imaging showed that the typical small-scale reticulated ridges on the upper side of L. pulmonaria caused a spatial, small-scale reduction in damage due to minor shading. Severe dry-state photoinhibition readily occurred in flattened and light-treated L. pulmonaria, although the mechanisms for such damage in a desiccated and inactive stage are not well known. Natural curling is one strategy to reduce the chance for serious photoinhibition in desiccated L. pulmonaria thalli during high light exposures.