Effects of drought on photosynthesis,chlorophyll fluorescence and photoinhibition susceptibility in intact willow leaves

Plants from clonal cuttings of Salix sp. were subjected to a drying cycle of 10 d in a controlled environment. Gas exchange and fluorescence emission were measured on attached leaves. The light-saturated photosynthetic CO₂ uptake became progressively inhibited with decreased leaf water potential both at high, and especially, at low intercellular CO₂ pressure. The maximal quantum yield of CO₂ uptake was more resistant. The inhibition of light-saturated CO₂ uptake at leaf water potentials around-10 bar, measured at a natural ambient CO₂ concentration, was equally attributable to stomatal and non-stomatal factors, but the further inhibition below this water-stress level was caused solely by non-stomatal factors. The kinetics of fluorescence emission was changed at severe water stress; the slow secondary oscillations of the induction curve were attenuated, and this probably indicates perturbations in the carbon reduction cycle. The influence of light level during the drought period was also studied. Provided the leaves were properly light-acclimated, drought at high and low light levels produced essentially the same effects on photosynthesis. However, low-light-acclimated leaves became more susceptible to photoinhibitory treatment under severe water stress, as compared with well-watered conditions.     

Evaluation of photosynthetic electron flow using simultaneous measurements of gas exchange and chlorophyll fluorescence under photorespiratory conditions

Simultaneous measurements of leaf gas exchange and chlorophyll fluorescence for Koelreuteria paniculata Laxm. at 380 ± 5.6 and 600 ± 8.5 ??mol mol^?1 were conducted, and the photosynthetic electron flow via photosystem II (PSII) to photosynthesis, photorespiration, and other electron-consuming processes were calculated. The results showed that the photosynthetic electron flow associated with carboxylation (J _c), oxygenation (J _o), and other electron-consuming processes (J _r) were 72.7, 45.7, and 29.4 ??mol(e^?) m^?2 s^?1 at 380 ??mol mol^?1, respectively; and 86.1, 35.3, and 48.2 ??mol(e^?) m^?2 s^?1 at 600 ??mol mol^?1, respectively. Our results revealed that other aspects associated with electronconsuming processes, except for photosynthesis and respiration, were neither negligible nor constant under photorespiratory conditions. Using maximum net photosynthetic rate (P _max), day respiration (R), photorespiration rate (R _l), and maximum electron flow via PSII (J _max), the use efficiency of electrons via PSII at saturation irradiance to fix CO₂ was calculated. The calculated results showed that the use efficiency of electrons via PSII to fix CO₂ at 600 ??mo^l mol^?1 was almost as effective as that at 380 ??mol mol^?1, even though more electrons passed through PSII at 600 ??mol mol^?1 than at 380 ??mol mol^?1.     

ApoE and clusterin cooperatively suppress Abeta levels and deposition: evidence that ApoE regulates extracellular Abeta metabolism in vivo

Apolipoprotein E (apoE) and clusterin can influence structure, toxicity, and accumulation of the amyloid-beta (Abeta) peptide in brain. Both molecules may also be involved in Abeta metabolism prior to its deposition. To assess this possibility, we compared PDAPP transgenic mice that develop age-dependent Abeta accumulation in the absence of apoE or clusterin as well as in the absence of both proteins. apoE(-/-) and clusterin(-/-) mice accumulated similar Abeta levels but much less fibrillar Abeta. In contrast, apoE(-/-)/clusterin(-/-) mice had both earlier onset and markedly increased Abeta and amyloid deposition. Both apoE(-/-) and apoE(-/-)/clusterin(-/-) mice had elevated CSF and brain interstitial fluid Abeta, as well as significant differences in the elimination half-life of interstitial fluid Abeta measured by in vivo microdialysis. These findings demonstrate additive effects of apoE and clusterin on influencing Abeta deposition and that apoE plays an important role in regulating extracellular CNS Abeta metabolism independent of Abeta synthesis.     

The influence of vertical mixing on the photoinhibition of variable chlorophyll a fluorescence and its inclusion in a model of phytoplankton photosynthesis

The maximum effective quantum yield of photosystem II was estimatedfrom measurements of variable chlorophyll a fluorescence [(F'_m- F'_o)/F'_m = F'_v/F'_m] in samples of phytoplankton collectedfrom various depths in Chaffey Reservoir, Australia. Duringstratified conditions, F'_v/F'_m showed depth-dependent decreasesas irradiance increased during the morning, and increases asirradiancereduced in the afternoon. Wind-induced mixing disrupted thediel pattern, but even under well-mixed conditions a verticalgradient in F'_v/F'_m remained. Differences in F'_v/F'_m valuesbetween samples incubated at fixed depths and unconstrainedlake samples enabled identification of the phytoplankton mixingdepth. Recovery of F'_v/F'_m was modelled as a function of timeand the degree of F'_v/F'_m inhibition, while damage was considereda function of photon dose. A combined, numerical model was fittedto diel sequences of F'_v/F'_m to estimate rate constants fordamage and repair. Recovery rate constants ranged from r = 0.7x 10^-4 to 9.1 x 10^-4 s^-1, while damage rate constants rangedfrom k = 0.03 to 0.22 m² mol photon^-1. A fluorescence-basedmodel of photosynthesis was used to investigate the effectsof wind speed, euphotic depth and mixed layer depth on photoinhibition.At different mixing rates, depth-integrated photosynthesis wasenhanced by up to 16% under the conditions tested, while increasesof 9% occurred between phytoplankton with different measureddamage and repair characteristics.     

Effects of chilling and high temperatures on photosynthesis and chlorophyll fluorescence in leaves of watermelon seedlings

The effects of chilling (CT, day/night temperatures of 12/10 °C, an irradiance of 250 μmol m^?2 s^?1), chilling combined with a low irradiance (CL, 12/10 °C, 80 μmol m^?2 s^?1), and a high temperature (HT, 42/40 °C, 250 μmol m^?2 s^?1) on chlorophyll content, chlorophyll fluorescence, and gas exchange were studied in two watermelon cultivars, ZJ8424 and YS01, differing in their resistance. The chlorophyll content, net photosynthetic rate (P_N), stomatal conductance (g_s), and transpiration rate (E) decreased substantially, whereas the intercellular CO₂ concentration (c_i) increased when the two watermelon cultivars were grown under these stresses. The photosynthetic parameters showed greater changes at chilling than at the high temperature, and the CL caused a more pronounced inhibition in PN compared with the CT. After 2 d exposure to the CT, YS01 had higher P_N, g_s, and E, but a lower ci compared with ZJ8424. The maximum efficiency of photosystem (PS) II photochemistry (F_v/F_m), effective quantum yield of PS II photochemistry (Φ_PSII), photochemical quenching (qP), and electron transport rate (ETR) decreased under the CT and CL but showed only a slight drop under the HT. All these stresses significantly increased non-photochemical quenching (NPQ). The CT brought more damage to the photosynthetic apparatus of leaves compared with the CL. In addition, after returning to normal conditions (25/15 °C, 250 μmol m^?2 s^?1) for 3 d, the photosynthetic parameters recovered to pre-stress levels in HT treated seedlings but not in CT treated seedlings. In conclusion, the low irradiance could help to alleviate the extent of photoinhibition of PS II photochemistry caused by chilling and cv. ZJ8424 was more sensitive to the extreme temperatures than cv. YS01.     

高温胁迫对水稻剑叶光合和叶绿素荧光特征的影响

采用盆栽方法在人工气候箱内对超级杂交稻组合"天优998"4个生育期(抽穗期、乳熟期、蜡熟期和完熟期)分别进行5个高温处理(最高温度分别设定为32、35、38、40和42℃,日较差为6℃,每个处理为期5d,每天2h,以自然条件下生长的水稻为对照),研究了高温胁迫对水稻剑叶光合和荧光特征的影响。结果表明:高温对水稻剑叶光合和荧光特征具有显著影响,而且因高温处理、生育期、参数的不同而有差异,温度越高影响越大;高温处理后水稻剑叶叶绿素含量(SPAD)、净光合速率(Pn)、气孔导度(Gs)下降,胞间CO2浓度(Ci)上升;光化学效率(Fv/Fm)、实际量子产量(ΦPSII)、表观电子传递速率(ETR)、光化学猝灭系数(qP)、PSⅡ光化学反应(P)下降,初始荧光(Fo)、非光化学猝灭系数(qN)、其他热耗散(E)上升;当最高温度大于35℃时,4个生育期的光合和荧光指标大多发生了显著变化,当最高温度大于38℃时,则大幅下降;抽穗期和乳熟期Pn、Gs下降显著、Ci上升显著,蜡熟期和完熟期SPAD下降显著,但高温下抽穗期、乳熟期水稻剑叶Fv/Fm的下降幅度和Fo的上升幅度比蜡熟期、完熟期大;高温胁迫对水稻剑叶Pn的影响比SPAD大,抽穗和乳熟期水稻剑叶Pn下降主要是由气孔因素引起,而蜡熟期和完熟期剑叶Pn的下降则主要是由非气孔因素所致。     

The winter-red-leaf syndrome in Pistacia lentiscus: evidence that the anthocyanic phenotype suffers from nitrogen deficiency, low carboxylation efficiency and high risk of photoinhibition

Recent evidence indicates that winter-red leaf phenotypes in the mastic tree (Pistacia lentiscus) are more vulnerable to chronic photoinhibition during the cold season relative to winter-green phenotypes occurring in the same high light environment. This was judged by limitations in the maximum quantum yield of photosystem II (PSII), found in previous studies. In this investigation, we asked whether corresponding limitations in leaf gas exchange and carboxylation reactions could also be manifested. During the cold (“red”) season, net CO₂ assimilation rates (A) and stomatal conductances (g_s) in the red phenotype were considerably lower than in the green phenotype, while leaf internal CO₂ concentration (Ci) was higher. The differences were abolished in the “green” period of the year, the dry summer included. Analysis of A versus Ci curves indicated that CO₂ assimilation during winter in the red phenotype was limited by Rubisco content and/or activity rather than stomatal conductance. Leaf nitrogen levels in the red phenotype were considerably lower during the red-leaf period. Consequently, we suggest that the inherently low leaf nitrogen levels are linked to the low net photosynthetic rates of the red plants through a decrease in Rubisco content. Accordingly, the reduced capacity of the carboxylation reactions to act as photosynthetic electron sinks may explain the corresponding loss of PSII photon trapping efficiency, which cannot be fully alleviated by the screening effect of the accumulated anthocyanins.     

Satellite chlorophyll fluorescence measurements reveal large‐scale decoupling of photosynthesis and greenness dynamics in boreal evergreen forests

Mid‐to‐high latitude forests play an important role in the terrestrial carbon cycle, but the representation of photosynthesis in boreal forests by current modelling and observational methods is still challenging. In particular, the applicability of existing satellite‐based proxies of greenness to indicate photosynthetic activity is hindered by small annual changes in green biomass of the often evergreen tree population and by the confounding effects of background materials such as snow. As an alternative, satellite measurements of sun‐induced chlorophyll fluorescence (SIF) can be used as a direct proxy of photosynthetic activity. In this study, the start and end of the photosynthetically active season of the main boreal forests are analysed using spaceborne SIF measurements retrieved from the GOME‐2 instrument and compared to that of green biomass, proxied by vegetation indices including the Enhanced Vegetation Index (EVI) derived from MODIS data. We find that photosynthesis and greenness show a similar seasonality in deciduous forests. In high‐latitude evergreen needleleaf forests, however, the length of the photosynthetically active period indicated by SIF is up to 6 weeks longer than the green biomass changing period proxied by EVI, with SIF showing a start‐of‐season of approximately 1 month earlier than EVI. On average, the photosynthetic spring recovery as signalled by SIF occurs as soon as air temperatures exceed the freezing point (2–3 °C) and when the snow on the ground has not yet completely melted. These findings are supported by model data of gross primary production and a number of other studies which evaluated in situ observations of CO₂ fluxes, meteorology and the physiological state of the needles. Our results demonstrate the sensitivity of space‐based SIF measurements to light‐use efficiency of boreal forests and their potential for an unbiased detection of photosynthetic activity even under the challenging conditions interposed by evergreen boreal ecosystems.     

Estimation of photosynthesis in cyanobacteria by pulse-amplitude modulation chlorophyll fluorescence: problems and solutions

Cyanobacteria are photosynthetic prokaryotes and widely used for photosynthetic research as model organisms. Partly due to their prokaryotic nature, however, estimation of photosynthesis by chlorophyll fluorescence measurements is sometimes problematic in cyanobacteria. For example, plastoquinone pool is reduced in the dark-acclimated samples in many cyanobacterial species so that conventional protocol developed for land plants cannot be directly applied for cyanobacteria. Even for the estimation of the simplest chlorophyll fluorescence parameter, F _v/F _m, some additional protocol such as addition of DCMU or illumination of weak blue light is necessary. In this review, those problems in the measurements of chlorophyll fluorescence in cyanobacteria are introduced, and solutions to those problems are given.     

Leaf phosphate status and photosynthesis in vivo: Changes in light scattering and chlorophyll fluorescence during photosynthetic induction in sugar beet leaves

Light scattering and chlorophyll fluorescence were measured in vivo for leaves of sugar beet plants cultured with low levels of phosphate (P). Light scattering during photosynthetic induction was markedly increased in low-P compared to control leaves. This effect was reversible, disappearing within 24 h after the P supply was increased. The fluorescence induction curves also exhibited significant and reversible differences between low-P and control leaves. The changes in light scattering and chlorophyll fluorescence correlated well with changes in the rate of photosynthesis in vivo. We suggest that the increase in light scattering during induction in low-P plants may be due to a decreased ability of the Calvin cycle to utilize assimilatory power generated photochemically.     

O2-dependent electron flow,membrane energization and the mechanism of non-photochemical quenching of chlorophyll fluorescence

Recent progress in chlorophyll fluorescence research is reviewed, with emphasis on separation of photochemical and non-photochemical quenching coefficients (q_P and q_N) by the saturation pulse method. This is part of an introductory talk at the Wageningen Meeting on The use of chlorophyll fluorescence and other non-invasive techniques in plant stress physiology. The sequence of events is investigated which leads to down-regulation of PS II quantum yield in vivo, expressed in formation of q_N. The role of O₂-dependent electron flow for pH- and q_N-formation is emphasized. Previous conclusions on the rate of pseudocyclic transport are re-evaluated in view of high ascorbate peroxidase activity observed in intact chloroplasts. It is proposed that the combined Mehler-Peroxidase reaction is responsible for most of the q_N developed when CO₂-assimilation is limited. Dithiothreitol is shown to inhibit part of q_N-formation as well as peroxidase-induced electron flow. As to the actual mechanism of non-photochemical quenching, it is demonstrated that quenching is favored by treatments which slow down reactions at the PS II donor side. The same treatments are shown to stimulate charge recombination, as measured via 50 s luminescence. It is suggested that also in vivo internal thylakoid acidification leads to stimulation of charge recombination, although on a more rapid time scale. A unifying model is proposed, incorporating reaction center and antenna quenching, with primary control of pH at the PS II reaction center, involving radical pair spin transition and charge recombination to the triplet state in a first quenching step. In a second step, triplet excitation is trapped by zeaxanthin (if present) which in its triplet excited state causes additional quenching of singlet excited chlorophyll.Abbreviations q_P coefficient of photochemical quenching - q_N coefficient of non-photochemical quenching - q_E coefficient of energy-dependent quenching - LED light emitting diode     

Primary sites of ozone-induced perturbations of photosynthesis in leaves: identification and characterization in Phaseolus vulgaris using high resolution chlorophyll fluorescence imaging

High resolution imaging of chlorophyll a fluorescence was used to identify the sites at which ozone initially induces perturbations of photosynthesis in leaves of Phaseolus vulgaris. Leaves were exposed to 250 and 500 nmol mol(-1) ozone at a photosynthetically active photon flux density of 300 micromol m(-2) s(-1) for 3 h. Images of fluorescence parameters indicated that large decreases in both the maximum and operating quantum efficiencies of photosystem II had occurred in cells adjacent to stomata in the upper, but not lower, leaf surfaces. However, this treatment did not produce any significant changes in the maximum or operating quantum efficiencies of photosystem II in the leaves when estimated from fluorescence parameters measured with a conventional, integrating fluorometer. The localized decreases in photosystem II photochemical efficiencies were accompanied by an increase in the minimal fluorescence level, which is indicative of photoinactivation of photosystem II complexes and a decrease in stomatal conductance. Perturbations of photochemical efficiencies were not observed in cells associated with all of the stomata on the upper leaf surface or within cells distant from the upper leaf surface. It is concluded that ozone penetrates the leaf through stomata and initially damages only cells close to stomatal pores.     

Effects of high temperature on photosynthesis, chlorophyll fluorescence, chloroplast ultrastructure, and antioxidant activities in fingered citron

Effects of heat stress on the photosynthesis system and antioxidant activities in Fingered citron (Citrus medica var. sarcodactylis Swingle) were investigated. Two-year-old Fingered citron plants were exposed to different temperature (28, 35, 40, and 45°C) for 6 h; then the photosynthetic capacity, chlorophyll fluorescence, chloroplast ultrastructure, and antioxidant activities in the leaves were evaluated. Exposure to 40 and 45°C for 6 h resulted in a significant decrease in the photosynthetic rate (P _n), carboxylation efficiency (CE), the maximal photochemical efficiency of photosystem II, and the light-saturated photosynthetic rate, which were related to the reduction of CO₂ assimilation, inactivation of photosystem II and photosynthetic electron transport. Moreover, transmission electron microscopy showed chloroplast ultrastructural alterations, including their swelling, matrix zone expanding, and lamella structure loosening. Furthermore, heat stress, especially at 45°C, caused oxidative damage resulted from ROS accumulation in Fingered citron leaves accompanied by increases in activities of superoxide dismutase, peroxidase, and catalase. However, exposure to 35°C for 6 h or 40°C for 4 h had no significant influence on the photosynthetic capacity at all. The results suggest that Fingered citron plants show no heat injury when temperature is below 40°C.     

Oxygen and electron flow in C4 photosynthesis: Mehler reaction, photorespiration and CO2 concentration in the bundle sheath

The photosynthetic linear electron transport rate in excess of that used for CO₂ reduction was evaluated in Sorghum bicolor Moench. [NADP-malic enzyme (ME)-type C₄ plant], Amaranthus cruentus L. (NAD-ME-type C₄ plant) and Helianthus annuus L. (C₃ plant) leaves at different CO₂ and O₂ concentrations. The electron transport rate (J _F) was calculated from fluorescence using the light partitioning factor (relative PSII cross-section) determined under conditions where excess electron transport was assumed to be negligible: low light intensities, 500 μmol CO₂ · mol⁻¹ and 2% O₂. Under high light intensities there was a large excess of J _F/4 at 10–100% O₂ in the C₃ plant due to photorespiration, but very little in sorghum and somewhat more in amaranth, showing that photorespiration is suppressed, more in the NADP-ME- and less in the NAD-ME-type species. It is concluded that when C₄ photosynthesis is limited by supply of atmospheric CO₂ to the C₄ cycle, the C₃ cycle becomes limited by regeneration of ribulose 1,5-bisphosphate (RuBP) which in turn limits RuBP oxygenase activity and photorespiration. The rate of excess electron transport over that consumed for CO₂ fixation in C₄ plants was very sensitive to the presence of O₂ in the gas phase, rapidly increasing between 0.01 and 0.1% O₂, and at 2% O₂ it was about two-thirds of that at 21% O₂. This shows the importance of the Mehler O₂ reduction as an electron sink, compared with photorespiration in C₄ plants. However, the rate of the Mehler reaction is still too low to fully account for the extra ATP which is needed in C₄ photosynthesis. Received: 8 November 1997 / Accepted: 26 December 1997     

Effect of long-term H2S fumigation on photosynthesis in spinach. Correlation between CO2 fixation and chlorophyll a fluorescence

Spinach plunts (Spinacia oleracea L. cv. Monosa) were exposed to air with and without 0.25 μl l^-1 H₂S. Effects of H₂S exposure for up to 18 days on photosynthesis, dark respiration and on chlorophyll a fluorescence were studied. Dark respiration was not affected by H₂S fumigation. Photosynthetic CO₂ fixation decreased linearly with time in both control and fumigated plants. The rate of decrease in CO₂ fixation was faster in the fumigated plants; after 14 days of exposure the fumigated plants showed a decrease in CO₂ fixation of 23%äs compared with the control plants. The H₂S-induced decrease in CO₂ fixation was accompanied by a decrease in quenching of the chlorophyll fluorescence. The most characteristic change in chlorophyll fluorescence was a decreased difference between maximum and steady-state fluorescence [(P-T)/P), suggesting a reduced efficiency in the use of photochemical energy in photosynthesis. Differences in CO₂ fixation were more pronounced whcn measured at high light intensity; the maximum rate of CO₂ fixation at light saturation decreased significantly with time in the H₂S-exposed plants; after 14 days of H₂S exposure a decrease of more than 70% was noted. The decrease in CO₂ fixation could not be attributed to a decreased chlorophyll content; on the contrary, chlorophyll content even slightly increased during fumigation. The initial increase in CO₂ fixation rate with increasing light intensity was also reduced by prolonged H₂S fumigation, indicating an effect of H₂S fumigation on photosynthetic electron transport. Finally, the phytotoxicity of H₂S is discusscd in relation to the H₂S-induced changes in photosynthetic CO₂ fixation and chlorophyll a fluorescence, and the effect of H₂S on leaf development observed in earlier studies.     

Investigations on the electron transfer processes in Photosystem II: New insights by chlorophyll fluorescence decay measurements with additional saturating light pulses

Electron transfer processes in leaves were investigated by chlorophyll fluorescence decay measurements. A fast chlorophyll fluorescence decay was observed in the intact state, reflecting normal electron transfer in Photosystem II. After treatment with DCMU a slow chlorophyll fluorescence decay was measured due to blocked electron transfer after the primary quinone Q_A. Additional saturating light pulses, one between each two measuring pulses, were used to completely reduce Q_A of the intact leaf: the chlorophyll fluorescence decay became similar to that of a DCMU treated leaf. A decreased electron donation rate to the reaction centre P680 was obtained after treatment with hydroxylamine. The intensity of the additional saturating light pulses was not sufficient to reduce all Q_A under this condition and only a small increase of the average chlorophyll fluorescence decay time occurred. Following our previous paper [Berg et al. (1997) Photosynthetica 34, in press], we investigated the effects of water stress with the additional saturating light pulses. An almost complete reduction of Q_A was possible after water stress started. A small, but systematic shortening of the slow chlorophyll fluorescence decay followed, up to a relative loss of leaf mass of 80%. At this time a rapid shortening of the chlorophyll fluorescence decay occurred, caused by an electron deficiency at the donor site of PS II. Additional saturating light pulses had no effects on the chlorophyll fluorescence decay any more, revealing a radiationless recombination between the reduced primary quinone Q and the oxidized reaction centre P680⁺.     

CFD simulation of an internal spin-filter: evidence of lateral migration and exchange flow through the mesh

In the present work Computational Fluid Dynamics (CFD) was used to study the flow field and particle dynamics in an internal spin-filter (SF) bioreactor system. Evidence of a radial exchange flow through the filter mesh was detected, with a magnitude up to 130-fold higher than the perfusion flow, thus significantly contributing to radial drag. The exchange flow magnitude was significantly influenced by the filter rotation rate, but not by the perfusion flow, within the ranges evaluated. Previous reports had only given indirect evidences of this exchange flow phenomenon in spin-filters, but the current simulations were able to quantify and explain it. Flow pattern inside the spin-filter bioreactor resembled a typical Taylor-Couette flow, with vortices being formed in the annular gap and eventually penetrating the internal volume of the filter, thus being the probable reason for the significant exchange flow observed. The simulations also showed that cells become depleted in the vicinity of the mesh due to lateral particle migration. Cell concentration near the filter was approximately 50% of the bulk concentration, explaining why cell separation achieved in SFs is not solely due to size exclusion. The results presented indicate the power of CFD techniques to study and better understand spin-filter systems, aiming at the establishment of effective design, operation and scale-up criteria.