Prompt chlorophyll fluorescence as a tool for crop phenotyping: an example of barley landraces exposed to various abiotic stress factors

The study examined photosynthetic efficiency of two barley landraces (cvs. Arabi Abiad and Arabi Aswad) through a prompt fluorescence technique under influence of 14 different abiotic stress factors. The difference in the behavior of photosynthetic parameters under the same stress factor in–between cv. Arabi Abiad and cv. Arabi Aswad indicated different mechanisms of tolerance and strategies for the conversion of light energy into chemical energy for both the landraces. This study confirmed the suitability of some chlorophyll fluorescence parameters as reliable biomarkers for screening the plants at the level of photosynthetic apparatus.     

Chlorophyll a fluorescence--A useful tool for the early detection of temperature stress in spring barley (Hordeum vulgare L.)

The photosynthetic activity of two Syrian barley (Hordeum vulgare L.) landraces, Arabi Abiad (A. Abiad) and Arabi Aswad (A. Aswad), grown under low- and high- temperature stresses, were studied by analyzing the measured chlorophyll fluorescence signals. Both the applied stresses influenced photosystem II (PSII) activity. However, the effects depend on the stress type and the duration of its application. Phenomenological parameters were shifted shortly after the application of both stresses, whereas fluorescence ratios and yield values were altered most significantly after 7 days of stress imposition. The earliest changes in PSII activity of both cultivars were observed in the case of high temperature treatment. The maximal quantum efficiency of the photosynthetic apparatus (F(v)/F(M)) did not alter after stress application. Therefore, we could not recommend this parameter for early detection of such stress. In contrast, the results from the present investigation strongly indicate that the most significantly changed chlorophyll a fluorescence parameters could be used as an efficient tool for the early diagnosis of temperature stress in barley.     

Nodule carbohydrate metabolism and polyols involvement in the response of Medicago sativa to salt stress

Alterations of plant growth, chlorophyll fluorescence parameters, nodule carbon metabolism and polyols concentration as result of salt stress were examined in alfalfa (Medicago sativa). Plants, in symbiosis with Sinorhizobium meliloti GR4 strain, were grown under controlled conditions for 35 days (DAS) and subjected to 150 mM of NaCl stress. Plant biomass (PDW) and nitrogen fixation rate (NFR) were markedly affected by salt stress conditions; the highest reductions of PDW (50%) and NFR (40%) were registered at 84 DAS and 56 DAS, respectively. In addition, salinity affected the chlorophyll fluorescence parameters, decreased initial chlorophyll fluorescence (F₀) and increased the optimum quantum yield of PSII (F_v/F_m ratio). The enzyme activities sucrose synthase activity and phosphoenolpyruvate carboxylase, responsible for the carbon supply to the bacteroids by the formation of dicarboxylates, were drastically inhibited by salinity, mainly at 56 DAS with the beginning of flowering. The content of total soluble sugars and proline increased under salt stress, and these concentrations were higher in nodule than in leaf. This last result suggests that the nodule is an organ specially protected in order to maintain its functioning, even under stress conditions. Besides, the content of myoinositol and pinitol in leaves and nodules changed with the plant growth stage and the saline treatment. Under salinity stress, the concentrations of pinitol in nodule were higher than in leaf, which supports the central function of this molecule in the adaptive response of nodules to salt stress. The increase of pinitol synthesis in nodule of M. sativa under salt stress could be one of the adaptive features used by the plant.     

Antagonism between elevated CO2, nighttime warming,and summer drought reduces the robustness of PSII performance to freezing events

Plant responses to warming, elevated CO₂, and changes in summer precipitation patterns involve complex interactions. In this study we aim to reveal the single factor responses and their interactive effects on photosystem II (PSII) performance during an autumn-to-winter period. The study was carried out in the CLIMAITE multifactor experiment, which includes the combined impact of elevated CO₂ (free air carbon enrichment; CO₂), warming (passive nighttime warming; T) and summer drought (rain-excluding curtains; D) in a temperate heath ecosystem. PSII performance was probed by the effective quantum yield in light, F_v′/F_m′, using the pulse amplitude methodology, and the total performance index, PI_total, which integrate changes of the chlorophyll-a fluorescence transient including the maximal quantum yield in darkness, F_v/F_m.Decreasing temperature during autumn linearly reduced PI_total, both in the wavy hair-grass, Deschampsia flexuosa, and in the evergreen dwarf shrub common heather, Calluna vulgaris, and following freezing events the PI_total and F_v′/F_m′ were reduced even more. Contrary to expected, indirect effects of the previous summer drought reduced PSII performance before freezing events, particularly in Calluna. In combinations with elevated CO₂ interactive effects with drought, D × CO₂ and warming, T × D × CO₂, were negatively skewed and caused the reduction of PSII performance in both species after occurrence of freezing events. Neither passive nighttime warming nor elevated CO₂ as single factors reduced PSII performance via incomplete cold hardening as hypothesized. Instead, the passive nighttime warming strongly increased PSII performance, especially after freezing events, and when combined with elevated CO₂ a strongly skewed positive T × CO₂ interactive effect was seen. This indicates that these plants take advantage of the longer growing season induced by the warming in elevated CO₂ until a winter frost period becomes permanent. However, if previously exposed to summer drought this positive effect reverses via interactive D × CO₂ and T × D × CO₂ effects immediately after freezing events, causing the full combination of TDCO₂ not to differ from the control.In a future warmer climate with high CO₂ and summer drought, the occurrence of freezing events thus seem highly decisive for reducing PSII performance in the autumn-to-winter period. Such a reduced robustness of PSII performance may be highly decisive for the magnitude of the late season photosynthetic carbon uptake and reduce the growing season length in these temperate heath plants.     

Quenching partitioning through light-modulated chlorophyll fluorescence: A quantitative analysis to assess the fate of the absorbed light in the field

Plants use a small part of the total absorbed light energy for net carboxylation, while the remaining amount is dissipated via alternative pathways involving thermal processes, fluorescence and non-carboxylation photochemistry in order to limit the formation of reactive oxygen species (ROS) and other photooxidative risks. The commonly used analysis of the Photosystem II (PSII) fluorescence signals gives qualitative information about absorbed light energy management by plants, but it is difficult to appreciate the relative contribution of each pathway in energy partitioning.This study reports the application of quenching partitioning through a chlorophyll fluorescence approach performed on peach leaves subjected to three different light intensities for four durations of exposure in absence of recovery from photo-damage. This methodology was compared with the P₇₀₀ redox kinetic method for determining the functional PSII fraction in leaves. In the absence of recovery processes the active PSII concentration decayed with an increase in photon exposure (the product of irradiance and the time of exposure), following an exponential pattern according to the reciprocity law. The photoprotective thermal dissipation (Φ_NPQ) was proportional to irradiance up to 30 min of photoinhibitory treatment. Afterwards Φ_NPQ was limited by the increasing competition for the absorbed energy re-emitted by the inactive PSII (Φ_NF). Φ_NF increased with the photon exposure dissipating up to 70% of the total incoming energy. The energy funnelled to photochemistry (Φ_PSII) decreased with increasing exposure time or intensity, becoming zero after 120 min of photoinhibitory treatment at the maximum irradiance (2100 μmol photon m⁻² s⁻¹). The relation between the fraction of energy dissipated by the inactive PSII (derived from the quenching partitioning) and the inactive PSII fraction (measured with the P₇₀₀ redox kinetic method) was linear.The quenching partitioning through light-modulated chlorophyll fluorescence is a useful tool to analyse plant energy management and gives also a reasonable estimation of the active PSII fraction. This methodology can easily be used in the field as measurements are rapid, non-destructive and detection devices are portable.     

Action spectra of photosystems II and I and quantum yield of photosynthesis in leaves in State 1

The spectral global quantum yield (Y_II, electrons/photons absorbed) of photosystem II (PSII) was measured in sunflower leaves in State 1 using monochromatic light. The global quantum yield of PSI (Y_I) was measured using low-intensity monochromatic light flashes and the associated transmittance change at 810 nm. The 810-nm signal change was calibrated based on the number of electrons generated by PSII during the flash (4 · O₂ evolution) which arrived at the PSI donor side after a delay of 2 ms. The intrinsic quantum yield of PSI (y_I, electrons per photon absorbed by PSI) was measured at 712 nm, where photon absorption by PSII was small. The results were used to resolve the individual spectra of the excitation partitioning coefficients between PSI (a_I) and PSII (a_II) in leaves. For comparison, pigment–protein complexes for PSII and PSI were isolated, separated by sucrose density ultracentrifugation, and their optical density was measured. A good correlation was obtained for the spectral excitation partitioning coefficients measured by these different methods. The intrinsic yield of PSI was high (y_I = 0.88), but it absorbed only about 1/3 of quanta; consequently, about 2/3 of quanta were absorbed by PSII, but processed with the low intrinsic yield y_II = 0.63. In PSII, the quantum yield of charge separation was 0.89 as detected by variable fluorescence F_v/F_m, but 29% of separated charges recombined (Laisk A, Eichelmann H and Oja V, Photosynth. Res. 113, 145–155). At wavelengths less than 580 nm about 30% of excitation is absorbed by pigments poorly connected to either photosystem, most likely carotenoids bound in pigment–protein complexes.     

Calcium protection of PS2 complex of Phaseolus coccineus from cadmium toxicity: In vitro study

The action of 10 and 20 mM Ca against harmful Cd effect on PS2 complex isolated from leaves of Phaseolus coccineus L. cv. Pi?kny Ja? was studied. The changes in fast chlorophyll a fluorescence induction kinetics and protein composition of PS2 complex were the symptoms of Cd toxicity and Ca protection of PS2 complex. Calcium applied at 10 mM concentration prevented F₀ reduction caused by the presence of 250–1000 μM Cd in the incubation mixture, but that of (the variable chlorophyll a fluorescence) F_v, F_m, F_v/F₀, and F_v/F_m only at 250 μM Cd. Ca concentration doubling in the incubation mixture resulted in complete overcoming the toxicity of 250–1000 μM Cd to F_v and F_m. However, the protection of F_v/F₀ and the photochemical efficiency of PS2 (F_v/F_m) from 1000 μM Cd was only partial even at 20 mM Ca. A protective effect of 10 mM Ca on D1, D2 and 17 kDa proteins was found in PS2 complex exposed to 250 μM Cd, and on 43 kDa protein in the complex treated with 500 μM Cd. However, 20 mM Ca counteracted the toxicity of 500 μM Cd to the 43, 47 and 17 kDa proteins, as well as the harmful effect of 1000 μM Cd on 47 and 17 kDa ones.     

Photosynthesis and activity of photosystem II in response to drought stress in Amur Grape (Vitis amurensis Rupr.)

The Amur Grape (Vitis amurensis Rupr.) cultivars ??shuangFeng?? and ??ZuoShanyi?? were grown in shelter greenhouse under natural sunlight and subjected to drought. Sap flow rate, net photosynthetic rate (P _N), and chlorophyll (Chl) fluorescence were measured on Amur Grape leaves subjected to different drought treatments. Significant decreases in P _N were associated with increasing intercellular CO₂ concentration (C _i), suggesting that the reduction in P _N was caused by nonstomatal limitation. Analysis of OJIP transients according to the JIP-test protocol revealed that specific (per PSII reaction center) energy fluxes for light absorption, excitation energy trapping and electron transport have significantly changed. The appearance of a pronounced K-step and J-step in polyphasic rise of fluorescence transient suggested the oxygen-evolving complex and electron transport were inhibited. Drought stress has relatively little effect on the parameter maximal quantum yield of PSII photochemistry (F_v/F_m), but the performance index (PIABS) is more sensitive in different drought treatment. There are cultivar differences in the response of PSII activity to drought, the photosynthetic apparatus of ??ZuoShanyi?? cultivar is more resistant to drought than that of ??ShuangFeng??, and JIP-test could be a useful indicator for evaluation and selection to drought tolerance.     

Comparison of thermotolerance of sun-exposed peel and shaded peel of ‘Fuji’ apple

The thermotolerance of the sun-exposed peel and the shaded peel of ‘Fuji’ apple (Malus domestica Borkh.) fruit was evaluated by measuring pigments, chlorophyll a fluorescence transients and O₂ evolution or uptake after exposure to 25, 35, 40, 42, 44, 46 or 48 °C for 30 min in the dark. A major effect of heat stress at 46–48 °C on the chlorophyll a fluorescence transients was the appearance of a very clear K step at 200–300 μs for both peel types. The K step was slightly more pronounced in the sun-exposed peel than in the shaded peel, suggesting that the resistance of oxygen-evolving complex to heat stress is slightly lower in the sun-exposed peel than in the shaded peel. Minimal fluorescence (F_O), relative to the value at 25 °C, increased to a greater extent in the shaded peel than in the sun-exposed peel after exposure to 46–48 °C, but the temperature dependencies of F_O changes were similar for both peel types. Maximum quantum yield of PSII (F_V/F_M) decreased to a similar extent in the sun-exposed peel and the shaded peel as temperature rose from 25 to 44 °C, but the sun-exposed peel reached slightly lower values at 46–48 °C. Correspondingly, gross O₂ evolution rate, relative to that at 25 °C, was also slightly lower in the sun-exposed peel than in the shaded peel at 46–48 °C. In response to heat stress, the ratio of Q_A-reducing reaction centers (RCs) to total RCs and the ratio of Q_B-reducing RCs to Q_A-reducing RCs decreased, but both of them decreased to lower values in the sun-exposed peel than in the shaded peel at 46–48 °C, indicating that the capacity of electron transfer between P₆₈₀⁺ and Q_B via Q_A was damaged to a greater extent in the sun-exposed peel than in the shaded peel. At each given temperature, dark respiration was similar between the two peel types. Overall, it appears that the exposure to higher surface temperature under high light does not make the sun-exposed peel more tolerant of heat stress than the shaded peel of apple fruit.     

Action of tenuazonic acid,a natural phytotoxin,on photosystem II of spinach

Tenuazonic acid (TeA) is a putative phytotoxin obtained from Alternaria alternata, the organism that can cause brown leaf spot disease of Crofton weed (Eupatorium adenophorum). It is demonstrated here that the tenuazonic acid inhibits the activity of photosystem II (PSII); the I₅₀-value is 48 μg mL^?1. Evidences from chlorophyll fluorescence show that tenuazonic acid interrupts electron transport between Q_A and Q_B on the acceptor side of PSII. It does not have an effect on the antenna pigments, the oxygen-evolving complex (OEC) at the donor side of PSII. On the basis of the fluorescence induction kinetics and competition experiments with [¹⁴C]atrazine, it is shown that tenuazonic acid does not share the same binding environment with atrazine despite their common action target: the Q_B-site. It is concluded that tenuazonic acid is a member of a novel class of PSII inhibitors.     

Ozone stress in Melissa officinalis plants assessed by photosynthetic function

Photosynthetic functions have been investigated in ozone stressed (200 ppb, 5 h) Melissa officinalis plants at the end of fumigation and 24 and 48 h after. Plants exhibited foliar injury and membrane permeability was significantly increased, indicating that there was membrane damage. After the end of treatment, CO₂ fixation capacity decreased and this lasted during the recovery period (until a maximum of −63% when compared to controls). These strong negative effects on photosynthetic ability were observed to be due both to stomatal and mesophyllic limitations, since stomatal conductance decreased (−23%) and intercellular CO₂ concentration significantly increased (+41%). Reduction in PSII efficiency is evidenced by (i) decrease of F_v/F₀ (−11.4%), indicating a partial inhibition at PSII donor side; (ii) significant correlation between the apparent electron transport rate through PSII and photosynthetic activity, suggesting that the O₃-induced effects are well established, as demonstrated by the development of leaf necrosis; (iii) increase in electrons required to fix one molecule of CO₂, showing a decrease in activity of photosynthetic enzymes and their ability to fix CO₂ in the presence of O₃; (iv) decrease of q_L, resulting in an increase in the PSII excitation pressure. On the other hand, a regulatory adjustment of PSII efficiency was highlighted by (i) higher value of q_NP, abling to counteract the negative effects of O₃ at chloroplast level because of their capacity to dissipate the excess of excitation energy; (ii) increase of the xanthophyll cycle pool size and DEPS index, showing a marked activation of photoprotective mechanisms. This represents an active response that M. officinalis initiates to cope with increased oxidative load.     

PSII photochemistry and carboxylation efficiency in Liriodendron tulipifera under ozone exposure

Liriodendron tulipifera is an important forest plant which is commonly used in urban environments as a shade tree. Young plants have been exposed (under controlled conditions) to 120 ppb of O₃ for 45 consecutive days (5 h d⁻¹). The aim of this investigation was to clarify if O₃ limits the physiological performance of L. tulipifera. In treated plants, dynamics related to membrane injury, gas exchange and chlorophyll a fluorescence leads to: (i) increase in lipid peroxidation (maximum value of +78% 15 days after the fumigation, compared to controls); (ii) reduction of photosynthetic activity (up to 66% 28 days after the exposure), twinned with a partial stomatal closure and a store of CO₂ in substomatal chambers; (iii) reduction in carboxylation efficiency (−11% at the end of exposure); (iv) damage to PSII, as demonstrated by the increase in the PSII excitation pressure (−57% 28 days after the treatment). On this basis, O₃ should be considered very harmful to L. tulipifera, although the reduction of total chlorophylls content and the activation of xanthophyll cycle take place in order to attempt to regulate light absorbed energy limiting oxidative damage.     

Effects of Zn2+ and niflumic acid on photosynthesis in Glycine soja and Glycine max seedlings under NaCl stress

The effects of two anion/Cl^? channel inhibitors, Zn²⁺ and niflumic acid (NA), on seedling photosynthetic and fluorescent parameters of two Glycine soja populations (salt-tolerant BB52; salt-sensitive N23227) and Glycine max cultivar (salt-tolerant Lee68) were studied and compared under salt stress. Treatments with Zn²⁺ and NA only (10, 20 μmol L^?1) were also imposed for comparisons. Results showed that, there were non-toxic and non-nutritional effects of Zn²⁺ and NA treatments alone on seed germination and seedling growth of soybeans. Under 150 mmol L^?1 NaCl for 6 d, leaf chlorophyll and carotenoid contents, net photosynthetic rate (P_n), stomatal conductance (G_s), intercellular CO₂ concentration (C_i), transpiration rate (Tr), and the maximum photochemical efficiency of photosystem II (PS II) (F_v/F_m) except the stomatal limitation (Ls) significantly decreased in three kinds of soybean seedlings when compared with their control plants. The NaCl stress plus additional 20 μmol L^?1 Zn showed an obvious enhancement of leaf chlorophyll and carotenoid contents, P_n, G_s, C_i and Tr, especially for the G. max cultivar Lee68, but the supplementation of 20 μmol L^?1 NA showed the reverse effects.     

Time course analysis of ABA and non-ionic osmotic stress-induced changes in water status,chlorophyll fluorescence and osmotic adjustment in Arabidopsis thaliana wild-type (Columbia) and ABA-deficient mutant (aba2)

In the present study, we investigated time course changes of water status including relative water content (RWC), leaf osmotic potential (Ψ_Π), stomatal conductance (g_s), proline (Pro), chlorophyll fluorescence (F_v/F_m) and total chlorophyll content in the Arabidopsis thaliana under PEG-induced drought stress after exogenous ABA treatment. To a better explanation for the role of ABA in the water status of A. thaliana to drought stress, wild-type (Columbia) and ABA-deficient mutant (aba2) of A. thaliana were used in the present study. Moreover, three weeks old Arabidopsis seedlings were applied exogenously with 50 μM ABA and exposed to drought stress induced by 40% PEG8000 (−0.73 MPa) for 6 h, 12 h and 24 h (hours). Our findings indicate that RWC of wild-type and aba2 started to decrease in the first 12 h and 6 h of PEG-induced drought stress, respectively. However, exogenous treatment of 50 μM ABA increased their RWC under drought stress. On the other hand, while Ψ_Π of both genotypes started to decrease in the first 6 h of drought stress, these declines in Ψ_Π were prevented by ABA treatment under stress throughout the experiment; it was more pronounced in aba2 at 24 h. While the highest increase in g_s was obtained in aba2 after 24 h stress, ABA-induced highest decrease in g_s was obtained in the same genotype during 12 h, as compared to PEG-treated group alone. On the other hand, Pro content increased in all treatment groups of ABA-deficient mutant aba2 at 12 h and 24 h. However, Pro content in ABA + PEG treated aba2 plants was higher than in PEG- and ABA-treated plants alone at the end of the 24 h. Drought stress decreased F_v/F_m and total chlorophyll contents of both genotypes while 50 μM ABA alleviated these reductions during drought stress, as compared to PEG stressed plants. On the other hand, 50 μM ABA treatment alone did not create any remarkable effect on F_v/F_m and total chlorophyll contents.These findings indicate that exogenous ABA showed an alleviative effect against damage of drought stress on relative water content, osmotic potential, stomatal conductance, proline, chlorophyll fluorescence and total chlorophyll content of both genotypes during 24 h of drought stress treatment.     

Partial protection of photosynthetic apparatus from UV-B-induced damage by UV-A radiation

Alteration in the photosynthetic apparatus of clusterbean (Cyamopsis tetraganoloba) cotyledons owing to UV-B irradiation in the absence or presence of UV-A radiation (UV-A + UV-B) during steady phase of its growth has been studied. UV-B radiation induces a decline in the photosynthetic pigments content and O₂ evolution along with a modification in the absorption spectra of chloroplasts. UV-A + UV-B irradiation moderately reverses these changes. The partial restoration of F_V/F_M value and other fluorescence transient parameters in UV-A + UV-B treated sample compared to that of UV-B treated one suggest that UV-A helps in developing a protective pathway against UV-B-induced impairment. UV-B-mediated alteration in S state transition of Mn cluster associated with oxygen evolving complex, as appeared from TL glow curves, is retrieved by UV-A radiation and Car is considered to negotiate against UV-B-induced damage of photosynthetic apparatus.     

Increased photosynthetic activities and thermostability of photosystem II with leaf development of elm seedlings (Ulmus pumila) probed by the fast fluorescence rise OJIP

Experiments were conducted to investigate the photosynthetic activity and thermostability of photosystem II (PSII) in elm seedling (Ulmus pumila) leaves from initiation to full expansion. During leaf development, photosynthesis, measured as CO₂ fixation, increased gradually and reached a maximum value when leaves were fully developed. In parallel with the increase of carbon assimilation, chlorophyll content increased. The chlorophyll a fluorescence measurements showed that the maximum quantum yield of PSII primary photochemistry (φ_po), the efficiency with which the energy of trapped excitons is converted into the electron transport beyond Q_A (Ψ_o) and the quantum yield of electron transport beyond Q_A (φ_Eo) increased gradually. The low light experiments confirmed these results independently. When subjected to heat stress, young leaves exhibited progressively lower φ_po and maximal fluorescence (F_m) values with considerably higher minimal fluorescence (F_o) than mature leaves, demonstrating that PSII in newly initiating leaves is more sensitive to heat stress. Further analysis revealed that PSII structure in newly initiating leaves showed a robust alteration under heat stress, which was reflected by the clear K phase in the OJIP curves. Therefore, we suggest that the enhanced thermostability of PSII in the case of leaf growth might be associated with an improvement of the stability of the oxygen-evolving complex (OEC) to heat stress during leaf development.     

Ecophysiological responses of water hyacinth exposed to Cr3+ and Cr6+

Due to its wide industrial use, chromium (Cr) is considered a serious environmental pollutant of aquatic bodies. In order to investigate the ecophysiological responses of water hyacinth [Eichhornia crassipes (Mart.) Solms] to Cr treatment, plants were exposed to 1 and 10 mM Cr₂O₃ (Cr³⁺) and K₂Cr₂O₇ (Cr⁶⁺) concentrations for two or 4 days in a hydroponic system. Plants exposed to the higher concentration of Cr⁶⁺ for 4 days did not survive, whereas a 2 days treatment with 1 mM Cr³⁺ apparently stimulated growth. Analysis of Cr uptake indicated that most of the Cr accumulated in the roots, but some was also translocated and accumulated in the leaves. However, in plants exposed to Cr⁶⁺ (1 mM), a higher translocation of Cr from roots to shoots was observed. It is possible that the conversion from Cr⁶⁺ to Cr³⁺, which immobilizes Cr in roots, was not total due to the presence of Cr⁶⁺, causing deleterious effects on gas exchange, chlorophyll a fluorescence and photosynthetic pigment contents. Chlorophyll a was more sensitive to Cr than chlorophyll b. Cr³⁺ was shown to be less toxic than Cr⁶⁺ and, in some cases even increased photosynthesis and chlorophyll content. This result indicated that the F_v/F₀ ratio was more effective than the F_v/F_m ratio in monitoring the development of stress by Cr⁶⁺. There was a linear relationship between qP and F_v/F_m. No statistical differences were observed in NPQ and chlorophyll a/b ratio, but there was a tendency to decrease these values with Cr exposure. This suggests that there were alterations in thylakoid stacking, which might explain the data obtained for gas exchanges and other chlorophyll a fluorescence parameters.     

CO2 assimilation,photosystem II photochemistry,carbohydrate metabolism and antioxidant system of citrus leaves in response to boron stress

Seedlings of Citrus grandis were fertilized every other days for 15 weeks with nutrient solution containing 0 (deficiency), 10 μM (control) or 500 μM (excess) H₃BO₃. CO₂ assimilation and chlorophyll (Chl) content decreased to a greater degree in B-deficient than in B-excess leaves, but photosynthetic enzyme activities were similarly decreased. Starch accumulated in B-deficient leaves, but not in B-excess ones. Chlorophyll a fluorescence transient showed that the positive L- and K-steps were more pronounced in B-excess than in B-deficient leaves. Maximum quantum yield of primary photochemistry (F_v/F_m), maximum variable fluorescence (F_v), oxygen-evolving complex (OEC) were less decreased in B-deficient than in B-excess leaves, whereas minimum fluorescence (F₀) was less increased in B-deficient leaves. Boron-deficient leaves displayed higher or similar antioxidant enzyme activities and higher ascorbate (AsA) and reduced glutathione (GSH) contents compared to B-excess leaves. Content of thiobarbituric acid (TBA) reactive compounds was less increased by B-deficiency than by B-excess. We conclude that B-deficient leaves are less damaged by oxidative stress than B-excess leaves due to their higher ability to scavenge reactive oxygen species. Both the donor (i.e. the OEC) and the acceptor sides of photosystem II were less photoinhibited by B-deficiency than by B-excess. The greater decrease in CO₂ assimilation and Chl content in B-deficient leaves may be caused by the excessive accumulation of starch. The reduction of CO₂ assimilation by B-excess is probably caused by a combination of factors such as oxidative damage, reduced photosynthetic enzyme activities and impaired electron transport capacity.     

Fluorescence kinetics of PSII crystals containing Ca2 + or Sr2 + in the oxygen evolving complex

Photosystem II (PSII) is the pigment–protein complex which converts sunlight energy into chemical energy by catalysing the process of light-driven oxidation of water into reducing equivalents in the form of protons and electrons. Three-dimensional structures from x-ray crystallography have been used extensively to model these processes. However, the crystal structures are not necessarily identical to those of the solubilised complexes. Here we compared picosecond fluorescence of solubilised and crystallised PSII core particles isolated from the thermophilic cyanobacterium Thermosynechococcus elongatus. The fluorescence of the crystals is sensitive to the presence of artificial electron acceptors (K₃Fe(CN)₃) and electron transport inhibitors (DCMU). In PSII with reaction centres in the open state, the picosecond fluorescence of PSII crystals and solubilised PSII is indistinguishable. Additionally we compared picosecond fluorescence of native PSII with PSII in which Ca² in the oxygen evolving complex (OEC) is biosynthetically replaced by Sr^2 +. With the Sr^2 + replaced OEC the average fluorescence decay slows down slightly (81 ps to 85 ps), and reaction centres are less readily closed, indicating that both energy transfer/trapping and electron transfer are affected by the replacement.     

Two functional sites of phosphatidylglycerol for regulation of reaction of plastoquinone QB in photosystem II

Functional roles of an anionic lipid phosphatidylglycerol (PG) were studied in pgsA-gene-inactivated and cdsA-gene-inactivated/phycobilisome-less mutant cells of a cyanobacterium Synechocystis sp. PCC 6803, which can grow only in PG-supplemented media. 1) A few days of PG depletion suppressed oxygen evolution of mutant cells supported by p-benzoquinone (BQ). The suppression was recovered slowly in a week after PG re-addition. Measurements of fluorescence yield indicated the enhanced sensitivity of Q_B to the inactivation by BQ. It is assumed that the loss of low-affinity PG (PG_L) enhances the affinity for BQ that inactivates Q_B. 2) Oxygen evolution without BQ, supported by the endogenous electron acceptors, was slowly suppressed due to the direct inactivation of Q_B during 10 days of PG depletion, and was recovered rapidly within 10 h upon the PG re-addition. It is concluded that the loss of high-affinity PG (PG_H) displaces Q_B directly. 3) Electron microscopy images of PG-depleted cells showed the specific suppression of division of mutant cells, which had developed thylakoid membranes attaching phycobilisomes (PBS). 4) Although the PG-depletion for 14 days decreased the chlorophyll/PBS ratio to about 1/4, florescence spectra/lifetimes were not modified indicating the flexible energy transfer from PBS to different numbers of PSII. Longer PG-depletion enhanced allophycocyanin fluorescence at 683 nm with a long 1.2 ns lifetime indicating the suppression of energy transfer from PBS to PSII. 5) Action sites of PG_H, PG_L and other PG molecules on PSII structure are discussed.