Involvement of betacyanin in chilling-induced photoinhibition in leaves of <Emphasis Type="Italic">Suaeda salsa</Emphasis>

Seeds of Suaeda salsa were cultured in dark for 3 d and betacyanin accumulation in seedlings was promoted significantly. Then the seedlings with accumulated betacyanin (C+B) were transferred to 14/10 h light/dark and used for chilling treatment 15 d later. Photosystem 2 (PS2) photochemistry, D1 protein content, and xanthophyll cycle during the chilling-induced photoinhibition (exposed to 5 °C at a moderate photon flux density of 500 μmol m⁻² s⁻¹ for 3 h) and the subsequent restoration were compared between the C+B seedlings and the control (C) ones. The maximal efficiency of PS2 photochemistry (F_v/F_m), the efficiency of excitation energy capture by open PS2 centres (F_v′/F_m′), and the yield of PS2 electron transport (Φ_PS2) of the C+B and C leaves both decreased during photoinhibition. However, smaller decreases in F_v/F_m, F_v′/F_m′, and Φ_PS2 were observed in the C+B leaves than in C ones. At the same time, the deepoxidation state of xanthophyll cycle, indicated by (A+Z)/(V+A+Z) ratio, increased rapidly but the D1 protein content decreased considerably during the photoinhibition. The increase in rate of (A+Z)/(V+A+Z) was higher but the D1 protein turnover was slower in C+B than C leaves. After photoinhibition treatment, the plants were transferred to a dim irradiation (10 μmol m⁻² s⁻¹) at 25 °C for restoration. During restoration, the chlorophyll (Chl) fluorescence parameters, D1 protein content, and xanthophyll cycle components relaxed gradually, but the rate and level of restoration in the C+B leaves was greater than those in the C leaves. The addition of betacyanins to the thylakoid solution in vitro resulted in similar changes of F_v/F_m, D1 protein content, and (A+Z)/(V+A+Z) ratio during the chilling process. Therefore, betacyanin accumulation in S. salsa seedlings may result in higher resistance to photoinhibition, larger slowing down of D1 protein turnover, and enhancement of non-radiative energy dissipation associated with xanthophyll cycle, as well as in greater restoration after photoinhibition than in the control when subjected to chilling at moderate irradiance.     

Photosynthetic acclimation to photon irradiance and its relation to chlorophyll fluorescence and carbon assimilation in the halotolerant green alga Dunaliella viridis

This work describes the long-term acclimation of the halotolerant microalga Dunaliella viridis to different photon irradiance, ranging from darkness to 1500 μmol m⁻² s⁻¹. In order to assess the effects of long-term photoinhibition, changes in oxygen production rate, pigment composition, xanthophyll cycle and in vivo chlorophyll fluorescence using the saturating pulse method were measured. Growth rate was maximal at intermediate irradiance (250 and 700 μmol m⁻² s⁻¹). The increase in growth irradiance from 700 to 1500 μmol m⁻² s⁻¹ did not lead to further significant changes in pigment composition or EPS, indicating saturation in the pigment response to high light. Changes in Photosystem II optimum quantum yield (F_v/F_m) evidenced photoinhibition at 700 and especially at 1500 μmol m⁻² s⁻¹. The relation between photosynthetic electron flow rate and photosyntetic O₂ evolution was linear for cultures in darkness shifting to curvilinear as growth irradiance increased, suggesting the interference of the energy dissipation processes in oxygen evolution. Carbon assimilation efficiencies were studied in relation to changes in growth rate, internal carbon and nitrogen composition, and organic carbon released to the external medium. All illuminated cultures showed a high capability to maintain a C:N ratio between 6 and 7. The percentage of organic carbon released to the external medium increased to its maximum under high irradiance (1500 μmol m⁻² s⁻¹). These results suggest that the release of organic carbon could act as a secondary dissipation process when the xanthophyll cycle is saturated. This revised version was published online in June 2006 with corrections to the Cover Date.     

Enhancement of susceptivity to photoinhibition and photooxidation in rice chlorophyll <Emphasis Type="Italic">b</Emphasis>-less mutants

Two rice chlorophyll (Chl) b-less mutants (VG28-1, VG30-5) and the respective wild type (WT) plant (cv. Zhonghua No. 11) were analyzed for the changes in Chl fluorescence parameters, xanthophyll cycle pool, and its de-epoxidation state under exposure to strong irradiance, SI (1 700 μmol m⁻² s⁻¹). We also examined alterations in the chloroplast ultrastructure of the mutants induced by methyl viologen (MV) photooxidation. During HI (0–3.5 h), the photoinactivation of photosystem 2 (PS2) appeared earlier and more severely in Chl b-less mutants than in the WT. The decreases in maximal photochemical efficiency of PS2 in the dark (F_v/F_m), quantum efficiency of PS2 electron transport (Φ_PS2), photochemical quenching (q_P), as well as rate of photochemistry (P_rate), and the increases in de-epoxidation state (DES) and rate of thermal dissipation of excitation energy (D_rate) were significantly greater in Chl b-mutants compared with the WT plant. A relatively larger xanthophyll pool and 78–83 % conversion of violaxanthin into antheraxanthin and zeaxanthin in the mutants after 3.5 h of HI was accompanied with a high ratio of inactive/total PS2 (0.55–0.73) and high 1–q_P (0.57–0.68) which showed that the activities of the xanthophyll cycle were probably insufficient to protect the photosynthetic apparatus against photoinhibition. No apparent difference of chloroplast ultrastructure was found between Chl b-less mutants and WT plants grown under low, LI (180 μmol m⁻² s⁻¹) and high, HI (700 μmol m⁻² s⁻¹) irradiance. However, swollen chloroplasts and slight dilation of thylakoids occurred in both mutants and the WT grown under LI followed by MV treatment. These typical symptoms of photooxidative damage were aggravated as plants were exposed to HI. Distorted and loose scattered thylakoids were observed in particular in the Chl b-less mutants. A greater extent of photoinhibition and photooxidation in these mutants indicated that the susceptibility to HI and oxidative stresses was enhanced in the photosynthetic apparatus without Chl b most likely as a consequence of a smaller antenna size.     

Response of xanthophyll cycle and chloroplastic antioxidant enzymes to chilling stress in tomato over-expressing glycerol-3-phosphate acyltransferase gene

Over-expression of chloroplastic glycerol-3-phosphate acyltransferase gene (LeGPAT) increased unsaturated fatty acid contents in phosphatidylglycerol (PG) of thylakoid membrane in tomato. The effect of this increase on the xanthophyll cycle and chloroplast antioxidant enzymes was examined by comparing wild type (WT) tomato with the transgenic (TG) lines at chilling temperature (4 °C) under low irradiance (100 μmol m⁻² s⁻¹). Net photosynthetic rate and the maximal photochemical efficiency of photosystem (PS) 2 (F_v/F_m) in TG plants decreased more slowly during chilling stress and F_v/F_m recovered faster than that in WT plants under optimal conditions. The oxidizable P700 in both WT and TG plants decreased during chilling stress under low irradiance, but recovered faster in TG plants than in the WT ones. During chilling stress, non-photochemical quenching (NPQ) and the de-epoxidized ratio of xanthophyll cycle in WT plants were lower than those of TG tomatoes. The higher activities of superoxide dismutase (SOD) and ascorbate peroxidase (APX) in TG plants resulted in the reduction of O₂ ^−· and H₂O₂ contents during chilling stress. Hence the increase in content of unsaturated fatty acids in PG by the over-expression of LeGPAT could alleviate photoinhibition of PS2 and PS1 by improving the de-epoxidized ratio of xanthophyll cycle and activities of SOD and APX in chloroplast.     

The roles of specific xanthophylls in light utilization

To evaluate the role of specific xanthophylls in light utilization, wild-type and xanthophyll mutant plants (npq1, npq2, lut2, lut2npq1 and lut2npq2) from Arabidopsis thaliana were grown under three different light regimes: 30 (low light, LL), 150 (medium light, ML) and 450 (high light, HL) μmol photons m⁻² s⁻¹. We studied the pigment content, growth rate, xanthophyll cycle activity, chlorophyll fluorescence parameters and the response to photoinhibition. All genotypes differed strongly in the growth rates and the resistance against photoinhibition. In particular, replacement of lutein (Lut) by violaxanthin (Vx) in the lut2npq1 mutant did not affect the growth at non-saturating light intensities (LL and ML), but led to a pronounced reduction of growth under HL conditions, indicating an important photoprotective role of Lut. This was further supported by a much higher sensitivity of all Lut-deficient plants to photoinhibition in comparison with the wild type. In contrast, replacement of Lut by zeaxanthin (Zx) in lut2npq2 led to a pronounced reduction of growth under all light regimes, most likely related to the permanent non-photochemical dissipation of excitation energy by Zx at Vx-binding sites and the destabilization of antenna proteins by binding of Zx to Lut-binding sites. The high susceptibility of lut2npq2 to photoinhibition in comparison with npq2 further indicated that the photoprotective function of Zx is abolished in the absence of Lut. Thus, it can be concluded from our work that neither Vx nor Zx is able to fulfil the essential photoprotective function at Lut-binding sites under in vivo conditions.     

Photoinhibition of photosynthesis in intact kiwifruit (Actinidia deliciosa) leaves: effect of growth temperature on photoinhibition and recovery

Intact leaves of kiwifruit (Actinidia deliciosa (A. Chev.) C.F. Liang et A.R. Ferguson) from plants grown in a range of controlled temperatures from 15/10 to 30/25°C were exposed to a photon flux density (PFD) of 1500 μmol·m⁻²·s⁻¹ at leaf temperatures between 10 and 25°C. Photoinhibition and recovery were followed at the same temperatures and at a PFD of 20 μmol·m⁻²·s⁻¹, by measuring chlorophyll fluorescence at 77 K and 692 nm, by measuring the photon yield of photosynthetic O₂ evolution and light-saturated net photosynthetic CO₂ uptake. The growth of plants at low temperatures resulted in chronic photoinhibition as evident from reduced fluorescence and photon yields. However, low-temperature-grown plants apparently had a higher capacity to dissipate excess excitation energy than leaves from plants grown at high temperatures. Induced photoinhibition, from exposure to a PFD above that during growth, was less severe in low-temperature-grown plants, particularly at high exposure temperatures. Net changes in the instantaneous fluorescence,F ₀, indicated that little or no photoinhibition occurred when low-temperature-grown plants were exposed to high-light at high temperatures. In contrast, high-temperature-grown plants were highly susceptible to photoinhibitory damage at all exposure temperatures. These data indicate acclimation in photosynthesis and changes in the capacity to dissipate excess excitation energy occurred in kiwifruit leaves with changes in growth temperature. Both processes contributed to changes in susceptibility to photoinhibition at the different growth temperatures. However, growth temperature also affected the capacity for recovery, with leaves from plants grown at low temperatures having moderate rates of recovery at low temperatures compared with leaves from plants grown at high temperatures which had negligible recovery. This also contributed to the reduced susceptibility to photoinhibition in low-temperature-grown plants. However, extreme photoinhibition resulted in severe reductions in the efficiency and capacity for photosynthesis.     

Effects of Cold-Hardening on Chilling-Induced Photoinhibition of Photosynthesis and on Xanthophyll Cycle Pigments in Sweet Pepper

Two cultivars of Capsicum annuum L. were acclimated for 5 d at sub-optimal temperature (14 °C) and irradiance of 250 µmol m^–2 s^–1. This cold-hardening resulted in some reduction in the extent of photoinhibition during an 8 h exposure to high irradiance at 4 °C. Obvious differences were observed between non-hardened leaves (NHL) and cold-hardened leaves (CHL) in the recovery under low irradiance at room temperature. The CHL of both cultivars recovered faster than NHL, especially during the initial fast phase of recovery. Compared with NHL, the total content of carotenoids (Cars), based on chlorophyll, Chl (a+b), and the proportions of xanthophyll cycle pigments referred to total Cars increased in CHL, mainly due to an increase of violaxanthin (V) + antheraxanthin (A) + zeaxanthin (Z) content per mol Chl (a+b). Faster development and a higher non-photochemical quenching (NPQ) of Chl fluorescence, related to a stronger deepoxidation of the larger xanthophyll cycle pool in NHL, could act as a major defence mechanism to reduce the formation of reactive oxygen species during severe chilling. This is suggested by higher content of Z or Z+A in photoinhibition as well as by its rapid decline during the initial fast phase of recovery. In contrast to the chilling-sensitive cv. 0004, the chilling-tolerant cv. 1141 did more easily acclimate its photosynthetic apparatus to low temperatures.     

Increasing leaf glutathione through stem feeding does not acclimate Eucalyptus camaldulensis seedlings towards high-light stress

Elevated foliar concentrations of glutathione (GSH) are a common stress response and potentially crucial in conferring increased stress tolerance. The present study addressed the following questions: can increased foliar GSH levels be achieved in the short term by applying a stem feeding technique to tree seedlings? If yes, will elevated GSH concentrations provide improved tolerance to the adverse effects of high-light stress? To this end Eucalyptus camaldulensis seedlings were stem fed a 5 mM GSH solution for 6–7 h before subjecting them to high-light exposure designed to induce photoinhibition. GSH in leaves was measured using a standard photometric method, and the effect of the high-light treatment was evaluated by the decrease in the optimum quantum efficiency of photosystem II (PSII) measured by chlorophyll fluorescence (F _v/F _m). Stem feeding GSH significantly increased GSH concentrations in the leaves up to 40% above control plants. Exposure to artificial high-light intensity for 3 h induced significant photoinhibition in leaves, measured by a 15% decrease in F _v/F _m. At the same time, photosynthesis and stomatal conductance measurements indicated that leaf physiology was not disrupted as a result of the stem feeding technique. However, we have no indication that elevated GSH increased tolerance; neither did it increase sensitivity of plants to high light-induced photoinhibition. This result was accompanied by maintained rates of photochemistry before and after light stress. Unlike previous GSH-related experiments increased tolerance by increasing the rate of photochemistry was not achieved in the present experiment.     

Thermotolerance and Related Antioxidant Enzyme Activities Induced by Heat Acclimation and Salicylic Acid in Grape (Vitis vinifera L.) Leaves

Thermotolerance and related antioxidant enzyme activities induced by both heat acclimation and exogenous salicylic acid (SA) application were studied in grapevine (Vitis vinifera L. cv. Jingxiu). Heat acclimation and exogenous SA application induced comparable changes in thermotolerance, ascorbic acid (AsA), glutathione (GSH), and hydrogen peroxide (H₂O₂) concentrations, and in activities of the antioxidant enzymes superoxide dismutase (SOD), peroxidase (POD), glutathione reductase (GR), ascorbic peroxidase (APX) and catalase (CAT) in grape leaves. Within 1 h at 38 °C, free SA concentration in leaves rose from 3.1 μg g⁻¹ FW to 19.1 μg g⁻¹ FW, then sharply declined. SA application and heat acclimation induced thermotolerance were related to changes of antioxidant enzyme activities and antioxidant concentration, indicating a role for endogenous SA in heat acclimation in grape leaves.     

Photosynthesis and photoinhibition in two differently coloured varieties of <Emphasis Type="Italic">Oxalis triangularis</Emphasis> — the effect of anthocyanin content

The purpose of this study was to clarify effects of anthocyanins on photosynthesis and photoinhibition in green and red leaves of Oxalis triangularis. Gas analysis indicated that green plants had the highest apparent quantum yield for CO₂ assimilation [0.051 vs. 0.031 μmol(CO₂) μmol⁻¹(photon)] and the highest maximum photosynthesis [10.07 vs. 7.24 μmol(CO₂) m⁻² s⁻¹], while fluorescence measurements indicated that red plants had the highest PSII quantum yield [0.200 vs. 0.143 μmol(e⁻) μmol⁻¹(photon)] and ETR_max [66.27 vs. 44.34 μmol(e⁻) m⁻² s⁻¹]. Red plants had high contents of anthocyanins [20.11 mg g⁻¹(DM)], while green plants had low and undetectable levels of anthocyanin. Red plants also had statistically significantly (0.05>p>0.01) lower contents of xanthophyll cycle components [0.63 vs. 0.76 mg g⁻¹(DM)] and higher activities of the reactive oxygen scavenging enzyme ascorbate peroxidase [41.2 vs. 10.0 nkat g⁻¹(DM)]. Anthocyanins act as a sunscreen, protecting the chloroplasts from high light intensities. This shading effect causes a lower photosynthetic CO₂ assimilation in red plants compared to green plants, but a higher quantum efficiency of photosystem II (PSII). Anthocyanins contribute to photoprotection, compensating for lower xanthophyll content in red plants, and red plants are less photoinhibited than green plants, as illustrated by the F_v/F_m ratio.     

Photosystem II photochemistry and physiological parameters of three fodder shrubs, <Emphasis Type="Italic">Nitraria retusa</Emphasis>, <Emphasis Type="Italic">Atriplex halimus</Emphasis> and <Emphasis Type="Italic">Medicago arborea</Emphasis> under salt stress

Nitraria retusa and Atriplex halimus (xero-halophytes) plants were grown in the range 0–800 mM NaCl while Medicago arborea (glycophyte) in 0–300 mM NaCl. Salt stress caused a marked decrease in osmotic potential and a significant accumulation of Na⁺ and Cl⁻ in leaves of both species. Moderate salinity had a stimulating effect on growth rate, net CO₂ assimilation, transpiration and stomatal conductance for the xero-halophytic species. At higher salinities, these physiological parameters decreased significantly, and their percentages of reduction were higher in A. halimus than in N. retusa whereas, in M. arborea they decreased linearly with salinity. Nitraria retusa PSII photochemistry and carotenoid content were unaffected by salinity, but a reduction in chlorophyll content was observed at 800 mM NaCl. Similar results were found in A. halimus, but with a decrease in the efficiency of PSII (F′v/F′m) occurred at 800 mM. Conversely, in M. arborea plants we observed a significant reduction in pigment concentrations and chlorophyll fluorescence parameters. The marked toxic effect of Na⁺ and/or Cl⁻ observed in M. arborea indicates that salt damage effect could be attributed to ions’ toxicity, and that the reduction in photosynthesis is most probably due to damages in the photosynthetic apparatus rather than factors affecting stomatal closure. For the two halophyte species, it appears that there is occurrence of co-limitation of photosynthesis by stomatal and non-stomatal factors. Our results suggest that both N. retusa and A. halimus show high tolerance to both high salinity and photoinhibition while M. arborea was considered as a slightly salt tolerant species.     

Adaptation of the photosynthetic apparatus of Nitellopsis obtusa to changing light intensity at the molecular level: different pathways of a singlet excitation quenching

The effect of prolonged illumination (60 min) with photosynthetically active monochromatic radiation of low intensity (3 μmol m⁻² s⁻¹) and high intensity (60 μmol m⁻² s⁻¹), corresponding to the physiological conditions and light stress conditions, respectively, was studied in the algae Nitellopsis obtusa. Illumination of Nitellopsis obtusa cells with strong light was associated with activation of the xanthophyll cycle, manifested by the deepoxidation of violaxanthin and accumulation of antheraxanthin and zeaxanthin. At the same time, the efficient singlet excitation quenching in the photosynthetic apparatus was activated, as demonstrated by the decrease in the intensity of the chlorophyll a fluorescence emission by ca 50 %. The difference of the fluorescence excitation spectra recorded before and after the light treatment match the difference absorption spectrum of the xanthophyll cycle pigments. The illumination with low light intensity resulted also in the chlorophyll a fluorescence quenching but the effect was very small (less than 10 %). The fluorescence quenching is interpreted in terms of the energy transfer between the Q_y energy level of chlorophyll a and the 2¹ A_g ⁻ energy level of zeaxanthin. The singlet energy levels of carotenoids, corresponding to the green spectral region, are also taken into consideration in the interpretation of the excitation energy exchange between the carotenoids and chlorophylls. Possible molecular mechanisms involved in the activation of the strong and the weak excitation quenching, including violaxanthin isomerization, and possible physiological functions of such pathways of energy transfer are discussed.     

Water stress and photoinhibition in acclimatization ofRosa hybrida plantlets

Summary MicropropagatedRosa hybrida plantlets were simultaneously rooted and acclimatized under 100 and 200 μmol m⁻² s⁻¹ light for 2 wk. At the end of the first week of acclimatization, the plantlets were transferred onto a low water potential medium (from −0.06 MPa to −0.3 MPa). Dry weight was decreased by increased hight and low water potential. Photoinhibition of photosynthesis, expressed as a decrease in Fv/Fm ratio and ΦPSII and an increase in 1 −qp, occurred in plants grown under 200 μmol m⁻² s⁻¹. When high light (200 μmol m⁻² s⁻¹) and water stress were applied simultaneously, their effects on chlorophyll fluorescence parameters depended on stress duration; after 1 d of water stress, photoinhibition was more pronounced; after 7 d of stress, Fv/Fm ratio and ΦPSII were higher than after 1 d of stress; photoinhibition was reduced. This suggests that after a 1-d stress, the effect of water stress alone included a superimposed effect of photoinhibition to which the water-stressed plants were sensitized; after 7 d, plantlets had adapted to water stress. The photoprotective effects under high light might result in energy dissipative mechanisms linked to photochemical and nonphotochemical quenching other than CO₂ fixation.     

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.     

Physiological and antioxidant responses of <Emphasis Type="Italic">Mentha pulegium</Emphasis> (Pennyroyal) to salt stress

Mentha pulegium L. is a medicinal and aromatic plant belonging to the Labiatae family present in the humid to the arid bioclimatic regions of Tunisia. We studied the effect of different salt concentrations on plant growth, mineral composition and antioxidant responses. Physiological and biochemical parameters were assessed in the plant organs after 2 weeks of salt treatment with 25, 50, 75 and 100 mM NaCl. Results showed that, growth was reduced even by 25 mM, and salt effect was more pronounced in shoots (leaves and stems) than in roots. This growth decrease was accompanied by a restriction in tissue hydration and K⁺ uptake, as well as an increase in Na⁺ levels in all organs. Considering the response of antioxidant enzymes to salt, leaves and roots reacted differently to saline conditions. Leaf and root guaiacol peroxidase activity showed an increase by different concentration of NaCl, but superoxide dismutase activity in the same organs showed a slight modification in NaCl-treated leaves and roots. Moreover, polyphenol contents and antioxidant activity were analysed in M. pulegium leaves and roots under salt constraint. The analysis showed an increase of total polyphenol content (2.41–8.17 mg gallic acid equivalent g⁻¹ dry weight) in leaves. However, methanol extract of leaves at 100 mM NaCl displayed the highest DPPH· scavenging ability with the lowest IC₅₀ value (0.27 μg ml⁻¹) in comparison with control which exhibited IC₅₀ equal to 0.79 μg ml⁻¹.     

Photosynthetic induction in leaves of co-occurring <Emphasis Type="Italic">Fagus lucida</Emphasis> and <Emphasis Type="Italic">Castanopsis lamontii</Emphasis> saplings grown in contrasting light environments

Photosynthetic induction times and photoinhibition in relation to simulated sunflecks (sudden increase of irradiance from 20 to 1,500 μmol m⁻² s⁻¹) were examined in leaves of co-occurring Fagus lucida (a deciduous tree) and Castanopsis lamontii (an evergreen tree) saplings grown either in a beech forest understory or in an adjacent open site during a late rainy season. Two hypotheses were tested: (1) understory leaves would display faster photosynthetic induction times and greater photoinhibition than open-grown leaves; and (2) evergreen species would have slower photosynthetic induction times and lighter photoinhibition than deciduous species. Times to reach 90% of maximal CO₂ assimilation rate (t _90%A ) and stomatal conductance did not differ between species, but showed faster by 3–5 min in open-grown leaves than understory leaves due to higher initial stomatal conductance (g _{s initial}) and induction state 1 min into simulated sunflecks (IS_1min) in the former. Our analysis across the published data on photosynthetic induction of 48 broad-leaved woody species again revealed the negative correlations between t _90%A and either g _{s initial} or IS_1min, and the similarity of t _90%A and between evergreen and deciduous species. Measurements of maximum PSII photochemical efficiency (F _v/F _m) indicated that photoinhibition occurred in saplings in any of the growth habitats during sunfleck-induced photosynthetic induction. Despite no interspecific differences in the degree of photoinhibition, understory leaves of both species suffered heavier photoinhibition than open-grown leaves, as indicated by a stronger decrease of F _v/F _m in the former. Dynamic changes in the quantum yields of PSII photochemistry and ΔpH- and xanthophyll-regulated thermal dissipation and adjustments in the partitioning of electron flow between assimilative and non-assimilative processes were functional to resist photoinhibition. However, such photoinhibition, together with stomatal and biochemical limitations, would decrease carbon gain during simulated sunflecks, particularly in understory leaves.     

Increase in Resistance to Low Temperature Photoinhibition Following Ascorbate Feeding is Attributable to an Enhanced Xanthophyll Cycle Activity in Rice (Oryza sativa L.) Leaves

The mechanistic basis for protection of exogenous ascorbate against photoinhibition at low temperature was examined in leaves of rice (Oryza sativa L.). Exposure of intact leaves to chilling temperature resulted in a drastic decrease in the speed of development of non-photochemical fluorescence quenching (NPQ). This was related to the low temperature-imposed restriction on the formation of the fast relaxing component of NPQ (q_f). Feeding with 20 mM ascorbate markedly increased the rate of q_f development at chilling temperature due primarily to the enhanced rate of zeaxanthin (Z) formation. On the other hand, ascorbate feeding had no influence on photosystem 2 (PS2)-driven electron flow. The reduced state of the PS2 primary electron acceptor Q_A decreased in ascorbate-fed leaves exposed to high irradiance at chilling temperature owing to the increased Z-associated thermal energy dissipation in the light-harvesting antenna system of PS2. Furthermore, ascorbate feeding increased the photosynthetic apparatus of rice leaves to resist photoinhibition at low temperature. The protective effect of exogenous ascorbate was fully accounted for by the enhanced xanthophyll cycle activity.     

Photoinhibition-induced reduction in photosynthesis is alleviated by abscisic acid,cytokinin and brassinosteroid in detached tomato leaves

Detached leaves of tomato (Lycopersicon esculentum Mill.) experienced photoinhibition associated with sharp reductions in net photosynthetic rate (Pn), quantum efficiency of PSII (Φ_PSII) and photochemical quenching (qP) even though they were exposed to mild light intensity (400 μmol m⁻² s⁻¹ PPFD) at 28°C. Photoinhibition and the reduction in Pn, Φ_PSII and qP, however, were significantly alleviated by 1 mg l⁻¹ ABA, 0.1 mg l⁻¹ N-(2-chloro-4-pyridyl)-N′-phenylurea (CPPU) and 0.01 mg l⁻¹ 24-epibrassinolide (EBR). Higher concentrations, however, reduced the effects or even exacerbated the occurrence of photoinhibition. Superoxide dismutase and ascorbate peroxidase activity in leaves increased with the increases in ABA concentration within 1–100 mg l⁻¹, CPPU concentration within 0.1–10 mg l⁻¹ and EBR concentration within 0.01–1.0 mg l⁻¹. Catalase and guaiacol peroxidase activity also increased with the increase in EBR concentration but CPPU and ABA treatments at higher concentrations caused a decrease. Malondialdehyde (MDA) content decreased with the increase in CPPU concentration. ABA and EBR, however, decreased MDA concentration only at 1 and 0.01 mg l⁻¹, respectively. In conclusion, detached leaves had increased sensitivity to PSII photoinhibition. Photoinhibition-induced decrease in photosynthesis, however, was significantly alleviated by EBR, CPPU and ABA at a proper concentration.     

Cooperation of xanthophyll cycle with water-water cycle in the protection of photosystems 1 and 2 against inactivation during chilling stress under low irradiance

The xanthophyll cycle and the water-water cycle had different functional significance in chilling-sensitive sweet pepper upon exposure to chilling temperature (4 °C) under low irradiance (100 µmol m⁻² s⁻¹) for 6 h. During chilling stress, effects of non-photochemical quenching (NPQ) on photosystem 2 (PS2) in dithiothreitol (DTT) fed leaves remained distinguishable from that of the water-water cycle in diethyldithiocarbamate (DDTC) fed leaves. In DTT-fed leaves, NPQ decreased greatly accompanied by visible inhibition of the de-epoxidized ratio of the xanthophyll cycle, and maximum photochemical efficiency of PS2 (F_v/F_m) decreased markedly. Thus the xanthophyll cycle-dependent NPQ could protect PS2 through energy dissipation under chilling stress. However, NPQ had a slighter effect on photosystem 1 (PS1) in DTT-fed leaves than in DDTC-fed leaves, whereas effects of the water-water cycle on PS1 remained distinguishable from that of NPQ. Inhibiting superoxide dismutase (SOD) activity increased the accumulation of , the oxidation level of P700 (P700⁺) decreased markedly relative to the control and DTT-fed leaves. Both F_v/F_m and NPQ changed little in DDTC-fed leaves accompanied by little change of (A+Z)/(V+A+Z). This is the active oxygen species inducing PS1 photoinhibition in sweet pepper. The water-water cycle can be interrupted easily at chilling temperature. We propose that during chilling stress under low irradiance, the xanthophyll cycle-dependent NPQ has the main function to protect PS2, whereas the water-water cycle is not only the pathway to dissipate energy but also the dominant factor causing PS1 chilling-sensitivity in sweet pepper.This research was supported by the State Key Basic Research and Development Plan of China (G1998010100), the Natural Science Foundation of China (30370854), and the open project from Key Lab of Crop Biology of Shandong Province.     

Dithiothreitol,an inhibitor of violaxanthin de-epoxidation,increases the susceptibility of leaves ofNerium oleander L. to photoinhibition of photosynthesis

Leaves ofNerium oleander L. plants, which had been previously kept in a shaded glasshouse for at least two months, were fed 1 mM dithiothreitol (DTT) through their petioles, either for 12h in darkness (overnight) or for 2h in low light (28 μmol photons·m⁻²·s⁻¹), in each case followed by a 3-h exposure to high light (1260 μmol photons·m⁻²·s⁻¹). During exposure to high light, violaxanthin became converted to zeaxanthin in control leaves, to which water had been fed, whereas zeaxanthin did not accumulate in leaves treated with DTT. Total carbon gain was not reduced by DTT during the photoinhibitory treatment. Exposure to high light led to a decrease in the photochemical efficiency of photosystem II, measured as the ratio of variable over maximum fluorescence emission,F _v/F _M, at both 298 K and 77K. The decrease was much more pronounced in the presence of DTT, mainly owing to a sustained increase in the instantaneous fluorescence,F _o. By contrast, in the control leaves,F _o determined immediately after the high-light treatment showed a transient decrease below theF _o value obtained before the onset of the photoinhibitory treatment (i.e. after 12 h dark adaptation), followed by a rapid return (within seconds) to this original level ofF _o during the following recovery period in darkness. Incubation of leaves with DTT led to large, sustained decreases in the photon-use efficiency of photosynthetic O₂ evolution by bright light, whilst the capacity of photosynthetic O₂ evolution at light and CO₂ saturation was less affected. In the control leaves, only small reductions in the photon yield and in the photosynthetic capacity were observed. These findings are consistent with previous suggestions that zeaxanthin, formed in the xanthophyll cycle by de-epoxidation of violaxanthin, is involved in protecting the photosynthetic apparatus against the adverse effects of excessive light.