The different effects of chilling stress under moderate light intensity on photosystem II compared with photosystem I and subsequent recovery in tropical tree species

Tropical plants are sensitive to chilling temperatures above zero but it is still unclear whether photosystem I (PSI) or photosystem II (PSII) of tropical plants is mainly affected by chilling temperatures. In this study, the effect of 4°C associated with various light densities on PSII and PSI was studied in the potted seedlings of four tropical evergreen tree species grown in an open field, Khaya ivorensis, Pometia tomentosa, Dalbergia odorifera, and Erythrophleum guineense. After 8 h chilling exposure at the different photosynthetic flux densities of 20, 50, 100, 150 μmol m⁻² s⁻¹, the maximum quantum yield of PSII (F _v /F _m) in all of the four species decreased little, while the quantity of efficient PSI complex (P _m) remained stable in all species except E. guineense. However, after chilling exposure under 250 μmol m⁻² s⁻¹ for 24 h, F _v /F _m was severely photoinhibited in all species whereas P _m was relative stable in all plants except E. guineense. At the chilling temperature of 4°C, electron transport from PSII to PSI was blocked because of excessive reduction of primary electron acceptor of PSII. F _v /F _m in these species except E. guineense recovered to ~90% after 8 h recovery in low light, suggesting the dependence of the recovery of PSII on moderate PSI and/or PSII activity. These results suggest that PSII is more sensitive to chilling temperature under the moderate light than PSI in tropical trees, and the photoinhibition of PSII and closure of PSII reaction centers can serve to protect PSI.     

Differential responses of photosystems I and II to seasonal drought in two Ficus species

Hemiepiphytic Ficus species exhibit more conservative water use strategy and are more drought-tolerant compared with their non-hemiepiphytic congeners, but a difference in the response of photosystem I (PSI) and photosystem II (PSII) to drought stress has not been documented to date. The enhancement of non-photochemical quenching (NPQ) and cyclic electron flow (CEF) have been identified as important mechanisms that protect the photosystems under drought conditions. Using the hemiepiphytic Ficus tinctoria and the non-hemiepiphytic Ficus racemosa, we studied the water status and the electron fluxes through PSI and PSII under seasonal water stress. Our results clearly indicated that the decline in the leaf predawn water potential (ψ_pd), the maximum photosynthetic rate (A_max) and the predawn maximum quantum yield of PSII (F_v/F_m) were more pronounced in F. racemosa than in F. tinctoria at peak drought. The F_v/F_m of F. racemosa was reduced to 0.69, indicating net photoinhibition of PSII. Concomitantly, the maximal photo-oxidizable P700 (P_m) decreased significantly in F. racemosa but remained stable in F. tinctoria. The fraction of non-photochemical quenching [Y(NPQ)] and the ratio of effective quantum yield of PSI to PSII [Y(I)/Y(II)] increased for both Ficus species at peak drought, with a stronger increase in F. racemosa. These results indicated that the enhancement of NPQ and the activation of CEF contributed to the photoprotection of PSI and PSII for both Ficus species under seasonal drought, particularly for F. racemosa.     

Cu2+ Inhibits Photosystem II Activities but Enhances Photosystem I Quantum Yield of Microcystis aeruginosa

Responses of photosystem I and II activities of Microcystis aeruginosa to various concentrations of Cu²⁺ were simultaneously examined using a Dual-PAM-100 fluorometer. Cell growth and contents of chlorophyll a were significantly inhibited by Cu²⁺. Photosystem II activity [Y(II)] and electron transport [rETR_max(II)] were significantly altered by Cu²⁺. The quantum yield of photosystem II [Y(II)] decreased by 29 % at 100 μg L^?1 Cu²⁺ compared to control. On the contrary, photosystem I was stable under Cu²⁺ stress and showed an obvious increase of quantum yield [Y(I)] and electron transport [rETR_max(I)] due to activation of cyclic electron flow (CEF). Yield of cyclic electron flow [Y(CEF)] was enhanced by 17 % at 100 μg L^?1 Cu²⁺ compared to control. The contribution of linear electron flow to photosystem I [Y(II)/Y(I)] decreased with increasing Cu²⁺ concentration. Yield of cyclic electron flow [Y(CEF)] was negatively correlated with the maximal photosystem II photochemical efficiency (F _v/F _m). In summary, photosystem II was the major target sites of toxicity of Cu²⁺, while photosystem I activity was enhanced under Cu²⁺ stress.     

Salinity improves chilling resistance in Suaeda salsa

Suaeda salsa L., a C3 euhalophytic herb, is native to saline soils, demonstrates high resistance to salinity stress. The effect of chilling stress on S. salsa under high salinity, particularly the change in unsaturated fatty acid content within membrane lipids, has not been investigated. After a 12 h chilling treatment (4 °C) performed under low irradiance (100 μmol m^?2 s^?1), the chlorophyll contents, maximal photochemical efficiency of photosystem II (F _v/F _m) and actual PSII efficiency (ΦPSII) were determined. These measurements were significantly decreased in S. salsa leaves in the absence of salt treatment yet there were no significant changes with a 200 mM NaCl treatment. Chlorophyll contents, F _v/F _m and ΦPSII in S. salsa under 200 mM NaCl were higher than those without salt treatment. The unsaturated fatty acid content and the double bond index (DBI) of major membrane lipids of monogalactosyldiacylglycerols, digalactosyldiacylglycerols (DGDG), sulphoquinovosyldiacylglycerols and phosphatidylglycerols (PG) significantly increased following the chilling treatment (4 °C) (with 12 h of low irradiance and 200 mM of NaCl). The DBI of DGDG and PG was decreased in the absence of the salt treatment. These results suggest that in the euhalophyte S. salsa, a 200 mM NaCl treatment increases chilling tolerance under conditions of low irradiance (100 μmol m^?2 s^?1).     

Fluorescence F 0 of photosystems II and I in developing C3 and C4 leaves,and implications on regulation of excitation balance

This work addresses the question of occurrence and function of photosystem II (PSII) in bundle sheath (BS) cells of leaves possessing NADP-malic enzyme-type C₄ photosynthesis (Zea mays). Although no requirement for PSII activity in the BS has been established, several component proteins of PSII have been detected in BS cells of developing maize leaves exhibiting O₂-insensitive photosynthesis. We used the basal fluorescence emissions of PSI (F _0I) and PSII (F _0II) as quantitative indicators of the respective relative photosystem densities. Chl fluorescence induction was measured simultaneously at 680 and 750 nm. In mature leaves, the F _m(680)/F ₀(680) ratio was 10.5 but less in immature leaves. We propose that the lower ratio was caused by the presence of a distinct non-variable component, F _c, emitting at 680 and 750 nm. After F _c was subtracted, the fluorescence of PSI (F _0I) was detected as a non-variable component at 750 nm and was undetectably low at 680 nm. Contents of Chls a and b were measured in addition to Chl fluorescence. The Chl b/(a + b) was relatively stable in developing sunflower leaves (0.25–0.26), but in maize it increased from 0.09 to 0.21 with leaf tissue age. In sunflower, the F _0I/(F _0I + F _0II) was 0.39 ± 0.01 independent of leaf age, but in maize, this parameter was 0.65 in young tissue of very low Chl content (20–50 mg m^?2) falling to a stable level of 0.53 ± 0.01 at Chl contents >100 mg m^?2. The values of F _0I/(F _0I + F _0II) showed that in sunflower, excitation was partitioned between PSII and PSI in a ratio of 2:1, but the same ratio was 1:1 in the C₄ plant. The latter is consistent with a PSII:PSI ratio of 2:1 in maize mesophyll cells and PSI only in BS cells (2:1:1 distribution). We suggest, moreover, that redox mediation of Chl synthesis, rather than protein accumulation, regulates photosystem assembly to ensure optimum excitation balance between functional PSII and PSI. Indeed, the apparent necessity for two Chls (a and b) may reside in their targeted functions in influencing accumulation of PSI and PSII, respectively, as opposed to their spectral differences.     

The interplay of anthocyanin biosynthesis and chlorophyll catabolism in senescing leaves and the question of photosystem II photoprotection

Fully exposed, senescing leaves of Cornus sanguinea and Parthenocissus quinquefolia display during autumn considerable variation in both anthocyanin and chlorophyll (Chl) concentrations. They were used in this study to test the hypothesis that anthocyanins may have a photoprotective function against photosystem II (PSII) photoinhibitory damage. The hypothesis could not be confirmed with field sampled leaves since maximum photochemical efficiency (F_v/F_m) of PSII was negatively correlated to anthocyanin concentration and the possible effects of anthocyanins were also confounded by a decrease in F_v/F_m with Chl loss. However, after short-term laboratory photoinhibitory trials, the percent decrease of F_v/F_m was independent of Chl concentration. In this case, a slight alleviation of PSII damage with increasing anthocyanins was observed in P. quinquefolia, while a similar trend in C. sanguinea was not statistically significant. It is inferred that the assumed photoprotection, if addressed to PSII, may be of limited advantage and only under adverse environmental conditions.     

Photosynthesis and biomass allocation of cotton as affected by deep-layer water and fertilizer application depth

Available water stored in deep soil layers could increase the photosynthetic capacity of cotton. It was hypothesized that the photosynthesis of cotton would be enhanced by changing the fertilizer application depth under different deep-layer water conditions. We examined two deep-layer water levels, i.e., well-watered (W₈₀) and not watered (W₀), combined with surface application (F₁₀) and deep application (F₃₀) of basal fertilizer. Compared to W₀, W₈₀ resulted in increased leaf area (LA), photosynthetic pigment contents, maximal PSII efficiency (F_v/F_m), effective quantum yield of PSII (Y_II) and PSI (Y_I), electron transport rate of PSII (ETR_II) and PSI (ETR_I). W₈₀ also increased the aboveground and root dry mass by 39 and 0.6%, respectively, and decreased the root/shoot ratio by 40–73%. Under the W₀ condition, higher values of F_v/F_m, Y_II, Y_I, ETR_II, and ETR_I were measured for F₁₀ compared to F₃₀ after 69 d from emergence. Under the W₈₀ condition, cotton plants with F₁₀ showed higher LA, F_v/F_m, Y_II, Y_I, ETR_II, and ETR_I, but there were no significant differences in the photosynthetic pigments compared to F₃₀. Our results suggest that sufficient water in deeper soil layers and the surface application of basal fertilizer could increase photosynthetic activity and efficiency, which promoted aboveground dry mass accumulation and partitioning towards reproductive organs.     

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.     

Photoprotective mechanisms in photosystem II of Ephedra monosperma during development of frost tolerance

Dissipation of light energy absorbed by photosystem II (PSII) in assimilating shoots of an evergreen shrub Ephedra monosperma was investigated during its transition from the vegetative to frost-tolerant state under natural conditions of Central Yakutia. The dynamics of modulated chlorophyll fluorescence and carotenoid content was analyzed during seasonal decrease in ambient temperature. The seasonal cooling was accompanied by a stepwise decrease in photochemical activity of PSII (F _v/F _m = (F _m ? F ₀)/F _m). The decrease in F _v/F _m occurred from the beginning of September to the end of October, when the temperature was lowered from 10 to ?8°C. During winter period the residual activity of PSII was retained at about 30% of the summer values. The seasonal decrease in temperature was accompanied by a significant stimulation of pH-independent dissipative processes in reaction centers and antenna of PSII. The increase in energy losses was paralleled by a proportional increase in zeaxanthin content on the background of decreasing content of violaxanthin and β-carotene as possible zeaxanthin precursors. At the same time, inhibition of light-induced non-photochemical quenching in the PSII antenna was observed. The results suggest that principal photoprotective mechanisms during seasonal lowering of temperature are: (1) inactivation of PSII and dissipation of excitation energy in PSII reaction centers and (2) zeaxanthin-mediated energy dissipation in the antenna complexes. The first mechanism seems to prevail at early stages of seasonal cooling, whereas both mechanisms are recruited from the onset of sustained freezing temperatures.     

Nutrient allocation and photochemical responses of <Emphasis Type="Italic">Populus</Emphasis> × <Emphasis Type="Italic">canadensis</Emphasis> ‘Neva’ to nitrogen fertilization and exogenous <Emphasis Type="Italic">Rhizophagus irregularis</Emphasis> inoculation

Arbuscular mycorrhizal fungi (AMF) can promote plant growth performance, but their effectiveness varies depending on soil nitrogen (N) availability. To clarify the effectiveness of exogenous AMF along an N-fertilization gradient (0, 2, 10, 20, and 30 mM), the impacts of exogenous Rhizophagus irregularis and N on the growth, photochemical activity, and nutritional status of Populus?×?canadensis ‘Neva’ in natural soil were evaluated in a pot experiment. The results showed that the 10 mM N level was the optimal fertilization regime with the highest promotion effect on plant growth and the maximum quantum yield of photosystem II (PSII) (F_v/F_m). Excess N (20 and 30 mM) fertilization reduced the actual quantum yield of PSII (ФPSII) and the F_v/F_m of the plants. Regardless of the N availability, inoculated plants exhibited greater F_v/F_m values than did non-inoculated plants. The biomass of inoculated plants was significantly higher compared with the control under low N levels (0 and 2 mM). Under high N levels, inoculated plants showed significant increases in ФPSII. Moreover, the nutrient imbalance of plants inoculated with exogenous R. irregularis was eased by increasing P, Fe, Mn and Cu uptake in roots and higher P, Ca, Mg, Fe, Mn and Zn concentrations in leaves. Moreover, the F_v/F_m and ФPSII exhibited positive correlations with P, Ca, Mg and Zn concentrations in leaves. In conclusion, inoculation with exogenous R. irregularis can benefit plant fitness by improving the photochemical capacity and nutrient composition of poplar under different N levels.     

Deriving fluorometer-specific values of relative PSI fluorescence intensity from quenching of F 0 fluorescence in leaves of Arabidopsis thaliana and Zea mays

The effect of stepwise increments of red light intensities on pulse-amplitude modulated (PAM) chlorophyll (Chl) fluorescence from leaves of A. thaliana and Z. mays was investigated. Minimum and maximum fluorescence were measured before illumination (F ₀ and F _M, respectively) and at the end of each light step ( $ F^{\prime}_{0} $ and $ F^{\prime}_{\text{M}} $ , respectively). Calculated $ F^{\prime}_{0} $ values derived from F ₀, F _M and $ F^{\prime}_{\text{M}} $ fluorescence according to Oxborough and Baker (1997) were lower than the corresponding measured $ F^{\prime}_{0} $ values. Based on the concept that calculated $ F^{\prime}_{0} $ values are under-estimated because the underlying theory ignores PSI fluorescence, a method was devised to gain relative PSI fluorescence intensities from differences between calculated and measured $ F^{\prime}_{0} $ . This method yields fluorometer-specific PSI data as its input data (F ₀, F _M, $ F^{\prime}_{0} $ and $ F^{\prime}_{\text{M}} $ ) depend solely on the spectral properties of the fluorometer used. Under the present conditions, the PSI contribution to F ₀ fluorescence was 0.24 in A. thaliana and it was independent on the light acclimation status; the corresponding value was 0.50 in Z. mays. Correction for PSI fluorescence affected Z. mays most: the linear relationship between PSI and PSII photochemical yields was clearly shifted toward the one-to-one proportionality line and maximum electron transport was increased by 50 %. Further, correction for PSI fluorescence increased the PSII reaction center-specific parameter, 1/F ₀ ? 1/F _M, up to 50 % in A. thaliana and up to 400 % in Z. mays.     

Evidence for leaf fold to remedy the deficiency of physiological photoprotection for photosystem II

An interesting phenomenon is that some light-demanding plants fold their leaves when exposed to high light. Since high light could induce selective photodamage to photosystem II (PSII), we suggest that the leaves fold themselves to diminish the absorption of light energy and remedy the deficiency of physiological photoprotection for PSII. To test this hypothesis, we determined light responses of non-photochemical quenching (NPQ) and cyclic electron flow (CEF) and the effect of high light on PSII activity in Microcos paniculata (non-foldable species) and Bauhinia tenuiflora (foldable species). Under high light B. tenuiflora showed much lower NPQ and CEF than M. paniculata. Meanwhile, the excess light energy that cannot be harmlessly dissipated in B. tenuiflora was more compared with that in M. paniculata. After exposure to a high light of 1,900 μmol photons m⁻² s⁻¹ for 2 h, the maximum quantum yield of PSII, as estimated by variable to maximal fluorescence (F _v /F _m) decreased from 0.7 to 0.52 in the foldable species B. tenuiflora but was stable at 0.7 in the nonfoldable species M. paniculata. These results indicate that the foldable species B. tenuiflora has more sensitivity of PSII to high light stress than the nonfoldable species M. paniculata, partly as a result of less CEF and NPQ in B. tenuiflora. Our results suggest that sun leaves fold themselves under high light to remedy the deficiency of physiological photoprotection for PSII.     

Gender, mediterranean drought, and seasonality: photosystem II photochemistry in Pistacia lentiscus L.

In this work, photosystem II (PSII) photochemistry, leaf water potential, and pigment contents of male and female Pistacia lentiscus L. were investigated during a seasonal cycle at three different, arid locations: superior semiarid, inferior semiarid, and arid. The results showed that the gender, season, and the site conditions interacted to influence the quantum yield and pigment contents in P. lentiscus. Predawn leaf water status was determined only by the site and season. The annual patterns of PSII maximum quantum efficiency (F_v/F_m) were characterized by a suboptimal activity during the winter, especially, populations with the more negative water potential exhibited a lower chlorophyll (Chl) a content and chronic photoinhibition irrespective of a gender. We also demonstrated that both photochemical or nonphotochemical mechanisms were involved to avoid the photoinhibition and both of them depended on the season. This plasticity of photosynthetic machinery was accompanied by changes in carotenoids and Chl balance. In the spring, the female F_v/F_m ratio was significantly higher than in male individuals, when the sexual dimorphism occurred during the fruiting stage, regardless of site conditions. P. lentiscus sex-ratio in Mediterranean areas, where precipitations exceeded 500 mm, was potentially female-biased. Among the fluorescence parameters investigated, nonphotochemical quenching coefficient appeared as the most useful one and a correlation was found between Chl a content and F_v/F_m. These results suggest that functional ecology studies would be possible on a large scale through light reflectance analysis.     

Cyclic electron flow plays an important role in photoprotection of tropical trees illuminated at temporal chilling temperature

Our previous study indicated that PSII is more sensitive to chilling and light stress than PSI in tropical trees, and Erythrophleum guineense is more sensitive to chilling stress than Dalbergia odorifera and Khaya ivorensis, but the underlying physiological mechanisms are unclear. Although recent studies have reported that cyclic electron flow (CEF) plays an important role in photoprotection, the role of CEF in protecting PSI and PSII of tropical tree species remains unclear. We investigated the effect of temporal chilling temperature on energy distribution in PSII, the redox state of P700 and CEF in the above-mentioned tropical evergreen tree species grown in an open field. Our results indicated that the overclosure of PSII reaction centers at chilling temperature led to excess excitation pressure in PSII. At the temporal chilling temperature under low light, PSI acceptor side limitation [Y(NA)] was lower than those at 25°C for all species. Although the effective quantum yield of CEF [Y(CEF)] was not significantly stimulated in E. guineense and K. ivorensis under temporal chilling at low light levels, the ratio of Y(CEF) to the effective quantum yield of PSII [Y(II)] significantly increased. Under chilling conditions Y(CEF)/Y(II) was stimulated much more in K. ivorensis and D. odorifera compared with that in the chilling-sensitive E. guineense. These results suggested that stimulation of Y(CEF)/Y(II) plays an important role in protecting PSI and PSII from photoinhibition caused by chilling stress.     

Changes in the photosynthetic characteristics and photosystem stoichiometries in wild-type and Chl <Emphasis Type="Italic">b</Emphasis>-deficient mutant rice seedlings under various irradiances

By using a wild-type rice (Oryza sativa L. cv. Norin No. 8) and the chlorophyll (Chl) b-deficient mutant derived from Norin No. 8 (chlorina 11), the present study monitored the oxygen evolution, contents of Chl a and b, β-carotene, and lutein in leaf and the contents of cytochrome f, and the reaction centres of photosystem I (PSI) and photosystem II (PSII) in thylakoids. The oxygen evolution, maximal quantum yield of PSII (F_v/F_m) and Chl concentration remained constant in both Norin No. 8 and chlorina 11 under 5 and 2% of full sunlight for six days. On the other hand, on the thylakoid level, the PSII reaction centre of chlorina 11 was more stable even under high irradiance, while approximately 40% decrease in levels of the PSII reaction centre occurred under 2% of full sunlight for six days. However, under such conditions, by regulating the stoichiometry of active PSII and PSI centres, the light absorption balance in both rice types was adjusted between the two photosystems. The present study attempted to examine whether the light absorption balance between PSII and PSI is altered to effectively conduct photosynthesis in the wild-type and Chl b-deficient mutant rice seedlings.     

Exogenous proline induces soluble sugar accumulation and alleviates drought stress effects on photosystem II functioning of <Emphasis Type="Italic">Arabidopsis thaliana</Emphasis> leaves

The effects of exogenous applied proline (Pro), on photosystem II (PSII) photochemistry of drought stressed (DS) 4-week old Arabidopsis thaliana plants, was studied by using chlorophyll (chl) fluorescence imaging. The maximum quantum yield of PSII photochemistry (F _v /F _m) in DS plants decreased significantly to 77% of that of the control value, suggesting that DS plants could not maintain PSII function, possibly due to accelerated photoinhibition of PSII. Free Pro and total soluble sugars (SS) increased, in response to DS. Exogenous foliar application of Pro by spraying, led to a remarkable increase in the accumulation of Pro and surprisingly also of SS. Both of them served to scavenge reactive oxygen species (ROS), as it was evident by the decreased lipid peroxidation level measured as malondialdehyde (MDA). DS plants sprayed with Pro showed a tolerance to photoinhibition, this indicated by F _v/F _m being close to values typical of healthy leaves by maintaining more than 98% of PSII function. Also the higher quantum efficiency of PSII photochemistry (Φ _PSΙΙ ) and the decreased excitation pressure (1 ? q _p ) recorded for stressed leaves with Pro, lead us to conclude that Pro appears to be involved in the protection of chloroplast structures by quenching ROS. The enhanced dissipation of excess light energy of PSII, in part accounts for the observed increased resistance to DS in A. thaliana leaves with Pro. Our data pointed out that Pro signalling interacts with SS signaling pathway and provided a new insight in Pro metabolism.     

Light-adapted charge-separated state of photosystem II: structural and functional dynamics of the closed reaction center

Photosystem II (PSII) uses solar energy to oxidize water and delivers electrons for life on Earth. The photochemical reaction center of PSII is known to possess two stationary states. In the open state (PSII_O), the absorption of a single photon triggers electron-transfer steps, which convert PSII into the charge-separated closed state (PSII_C). Here, by using steady-state and time-resolved spectroscopic techniques on Spinacia oleracea and Thermosynechococcus vulcanus preparations, we show that additional illumination gradually transforms PSII_C into a light-adapted charge-separated state (PSII_L). The PSII_C-to-PSII_L transition, observed at all temperatures between 80 and 308 K, is responsible for a large part of the variable chlorophyll-a fluorescence (F_v) and is associated with subtle, dark-reversible reorganizations in the core complexes, protein conformational changes at noncryogenic temperatures, and marked variations in the rates of photochemical and photophysical reactions. The build-up of PSII_L requires a series of light-induced events generating rapidly recombining primary radical pairs, spaced by sufficient waiting times between these events—pointing to the roles of local electric-field transients and dielectric relaxation processes. We show that the maximum fluorescence level, F_m, is associated with PSII_L rather than with PSII_C, and thus the F_v/F_m parameter cannot be equated with the quantum efficiency of PSII photochemistry. Our findings resolve the controversies and explain the peculiar features of chlorophyll-a fluorescence kinetics, a tool to monitor the functional activity and the structural-functional plasticity of PSII in different wild-types and mutant organisms and under stress conditions.

The closed-state of photosystem II possesses a hitherto unrecognized structural and functional plasticity and upon illumination assumes a light-adapted charge-separated state.     

Differences in the stimulation of cyclic electron flow in two tropical ferns under water stress are related to leaf anatomy

Cyclic electron flow (CEF) plays an important role in photoprotection for angiosperms under environmental stresses. However, ferns are more sensitive to drought and their water transport systems are not as efficient as those of angiosperms, it is unclear whether CEF also contributes to photoprotection in these plants. Using Microsorum punctatum and Paraleptochillus decurrens, we studied the electron fluxes through both photosystem I (PSI) and photosystem II (PSII) under water stress and their leaf anatomies. Our goal was to determine if CEF functions in the photoprotection of these ferns and, if so, whether CEF stimulation is related to leaf anatomy. Compared with P. decurrens, M. punctatum had thicker leaves and cuticles and higher water storage capacity, but lower stomatal density and slower rate of water loss. During induced drought, the decrease in leaf water potential (Ψ_leaf) was more pronounced in P. decurrens than in M. punctatum. For both species, the decline in Ψ_leaf was associated with a lower effective PSII quantum yield, photochemical quantum yield of PSI and electron transport rate (ETR), whereas increases were found in the quantum yield of regulated energy dissipation, CEF and CEF/ETR(II) ratio. Values for CEF and the CEF/ETR(II) ratio peaked in M. punctatum at a light intensity of 500–600 µmol m^?2 s^?1 vs only 150–200 µmol m^?2 s^?1 in P. decurrens. Therefore, our results indicate that the stimulation of CEF in tropical ferns contributes to their photoprotection under water stress, and is related to their respective drought tolerance and leaf anatomy.     

Analysis of dark-relaxation kinetics of variable fluorescence in intact leaves

The dark-relaxation kinetics of variable fluorescence, F_v, in intact green leaves of Pisum stativum L. and Dolichos lablab L. were analyzed using modulated fluorometers. Fast (t_1/2 = 1 s) and slow (t_1/2 = 7–8 s) phases in f_v dark-decay kinetics were observed; the rate and the relative contribution of each phase in total relaxation depended upon the fluence rate of the actinic light and the point in the induction curve at which the actinic light was switched off. The rate of the slow phase was accelerated markedly by illumination with far-red light; the slow phase was abolished by methyl viologen. The halftime of the fast phase of F_v dark decay decreased from 250 ms in dark-adapted leaves to 12–15 ms upon adaptation to red light which is absorbed by PSII. The analysis of the effect of far-red light, which is absorbed mainly by PSI, on F_v dark decay indicates that the slow phase develops when a fraction of Q_A ^– (the primary stable electron acceptor of PSII) cannot transfer electrons to PSI because of limitation on the availability of P700⁺ (the primary electron donor of PSI). After prolonged illumination of dark-adapted leaves in red (PSII-absorbed) light, a transient. F_v rise appears which is prevented by far-red (PSI-absorbed) light. This transient f_v rise reflects the accumulation of Q_A ^– in the dark. The observation of this transient F_v rise even in the presence of the uncoupler carbonylcyanide m-chlorophenyl hydrazone (CCCP) indicates that a mechanism other than ATP-driven back-transfer of electrons to QA may be responsible for the phenomenon. It is suggested that the fast phase in F_v dark-decay kinetics represents the reoxidation of Q_A ^– by the electron-transport chain to PSI, whereas the slow phase is likely to be related to the interaction of Q_A ^– with the donor side of PSII.Abbreviations CCCP carbonylcyanide m-chlorophenylhydrazone - F_O initial fluorescence level - F_v variable fluorescence - P700 primary electron donor of PSI - PSI, II photosystem I, II - Q_A (Q_A ^–) Q_B (Q_B ^–) primary and secondary stable electron acceptor of PSII in oxidized (reduced) state Supported by grant B6.1/88 DST, Govt. of India.