Response of photosynthetic machinery of field-grown kiwifruit under Mediterranean conditions during drought and re-watering

Groups of Actinidia deliciosa A. Chev. C.F. Liang et A.R. Ferguson var. deliciosa kiwifruit plants were subjected to soil water shortage (D), while other groups were well irrigated (I). Variations in chlorophyll (Chl) a fluorescence indices and leaf gas exchange were determined once plants were severely stressed (25 d after the beginning of the D-cycle). Daily maximum values of photosynthetic photon flux density (PPFD) were ca. 1 650 μmol(photon) m⁻² s⁻¹, while air temperatures peaked at 34.6 °C. High irradiance per se did not greatly affect the efficiency of photosystem (PS) 2, but predisposed its synergistic reduction by D co-occurrence. Fluorescence showed transient photodamage of PS2 with a complete recovery in the afternoon in both D and I plants. Upon re-watering the efficiency of PS2 was suboptimal (95 %) at day 2 after irrigation was reinitiated. At early morning of the day 5 of re-watering, photosynthesis and stomatal conductance recovered at about 95 and 80 % of I vines, respectively, indicating some after-stress effect on stomatal aperture. Once excessive photons reached PS2, the thermal dissipation of surplus excitation energy was the main strategy to save the photosynthetic apparatus and to optimize carbon fixation. The rather prompt recovery of both Chl a fluorescence indices and net photosynthetic rate during re-watering indicated that kiwifruit photosynthetic apparatus is prepared to cope with temporary water shortage under Mediterranean-type-climates.     

Thermostability of photosynthesis in two new chlorophyllb-less rice mutants

Leaves of the two new chlorophyll b-less rice mutants VG28-1, VG30-5 and the wild type rice cv. Zhonghua 11 were subjected to temperatures 28, 36, 40, 44 and 48℃ in the dark for 30 min or gradually elevated temperature from 30℃ to 80℃ at 0.5℃/min. The thermostability of photosynthetic apparatus was estimated by the changes in chlorophyll fluorescence parameters, photosynthetic rate and pigment content, chloroplast ultrastructure and tissue location of H2O2 accumulation. There were different patterns of Fo-temperature curves between the Chl b-less mutants and the wild type plant, and the temperature of F_o rising threshold was shifted 3℃ lower in the Chl b-less mutants (48℃) than in the wild type (51℃). At temperature up to about 45℃, chloroplasts were swollen and thylakoid grana became misty accompanied with the complete loss of photosynthetic oxygen evolution in the two Chl b-less mutants, but chloroplast ultrastruc-ture in the wild type showed no obvious alteration. After 55℃ exposure, the disordered thylakoid and significant H₂O₂ accumulation in leaves were found in the two Chl _b-less mutants, whereas in the wild type plant, less H₂O₂ was accumulated and the swollen thylakoid still maintained a cer-tain extent of stacking. A large extent of the changes in qP, NPQ and F_v/F_m was consistent with the Pn decreasing rate in the Chl b-less mutants during high temperature treatment as compared with the wild type. The results indicated that the Chl b-less mutants showed a tendency for higher thermosensitivity, and loss of Chl b in LHC II could lead to less thermostability of PSII structure and function. Heat damage to photosynthetic apparatus might be partially attributed to the in-ternal oxidative stress produced at severely high temperature.     

Effects of copper on the growth, photosynthesis and nutrient concentrations of Phaseolus plants

Bean plants (Phaseolus vulgaris L. var. Zargana Kavala) were grown under conditions of increasing Cu concentrations in the growth medium (0.5-160.5 μM). Generally, the Cu concentrations between 0.5-1.5 μM were deficient, 1.5-10.5 μM were optimal, and 10.5-160.5 μM were toxic to plant growth. The Cu toxicity was associated with marked increases in plant tissue Cu concentrations. Under the Cu-deficient and optimal growth conditions, Cu was located primarily in the leaves. Under Cu toxicity, it was primarily sequestered in the roots. With increasing Cu in the growth medium, there was a positive correlation between Cu concentrations in the roots, stems and leaves, Ca in the roots, and K and Mg in the leaves. In contrast, Ca concentrations in the leaves and stems showed a negative correlation. The chlorophyll (Chl) concentration increased with increasing leaf Cu concentration, however, the Chl a/b ratio decreased. Since with an increasing leaf Cu concentration the leaf area decreased more markedly than the leaf dry mass, the net photosynthetic rate (PN) per leaf area increased and per dry mass decreased. The increase in PN per leaf area was almost entirely accounted for by the increase in Chl concentration. The initial Chl fluorescence (F0) increased with increasing leaf Cu concentration. The ratio of variable to maximum fluorescence (Fv/Fm) under Cu toxicity decreased. The half-time for the rise from F0 to Fm (t1/2) remained relatively unchanged with increasing leaf Cu concentration. Therefore the Cu-stress caused a small decrease in the efficiency of photosystem 2 photochemistry, but its primary effect was on growth. This revised version was published online in June 2006 with corrections to the Cover Date.     

Isolation and Characterization of Photosystem Ⅱ of Porphyra yezoensis Ueda

The thylakoid membranes were isolated and purified from gametophyte of Porphyra yezoensis Ueda (P. yezoensis) by sucrose density gradient ultracentrifugation. After P. yezoensis gametophyte thylakoid membranes were solubilized with SDS, the photosystem Ⅱ (PSⅡ) particles were isolated and purified. The activity of PSⅡ     

Effects of Nutrient Limitation on Pigments in Thalassiosira weissflogii and Prorocentrum donghaiense

The response of Prorocentrum donghaiense and Thalassiosira weissfiogii pigments under nitrate （N） and phosphate （P） limitation were studied using HPLC and in vivo fluorescence protocols in batch cultures. For P. donghaiense, the pigment ratio was kept stable under different nutrient conditions from the results of HPLC. For T. weissflogii, there was a lower ratio of chlorophyllide to Chl a during the exponential phase, but the reverse during the stationary phase. Different members of the phytoplankton had different pigments response mechanisms under nutrient limitation. From the results of in vivo fluorescence, the ratio of peridinin to Chl a for P. donghaiense increased in nutrient-free culture, while it was kept stable for nutrient-limited cultures during the exponential phase. For T. weissflogii, the ratio of fucoxanthin to Chl a for each culture increased during the exponential phase, but the ratio under N limitation was apparently lower than that for P limitation during the stationary phase. The results indicate that both pigment ratios from HPLC and in vivo fluorescence of To weissflogii were changed greatly under different nutrient conditions, which suggests that both ratios could be used as indicators of algal physiological status in different nutrient conditions.     

Effects of copper and cadmium on photosynthesis in cucumber cotyledons

The effects of 20 and 50 μM concentrations of Cu and Cd on photosynthesis in cucumber (Cucumis sativus L.) cotyledons were studied by the measurements of gas exchange characteristics, chlorophyll (Chl) fluorescence parameters, photosynthetic pigment contents, and two Calvin cycle enzymes activities: glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and 3-phosphoglyceric acid kinase (PGK). To minimize indirect metal action, seedlings were treated with metals in the stage of green, fully developed cotyledons. The metals reached the cotyledon tissue after 48 h of treatments, though symptoms of metal action were not visible at that time. The effect of metals on the light phase of the photosynthesis parameters such as potential efficiency of photosystem 2 (PS2; F_v/F_m), and photochemical and nonphotochemical quenching of Chl fluorescence (q_P and q_NP) was negligible. In contrast, a decrease of PS2 quantum efficiency (Φ_PS2) was much more noticeable. Changes in the pigment contents were slight, as only 50 μM Cd decreased Chl a and b contents in small extent. On the contrary, metals in both concentrations drastically decreased (50 and more % of control) the net photosynthetic rate and the stomatal conductance, but not the internal CO₂ concentration. The activities of both GAPDH and PGK were also decreased by metals, although the effect on PGK was more prominent, particularly on its potential activity (dithiothreitol in extraction and incubation media). Hence Cu and Cd affected the synthesis of enzyme proteins rather than they influenced their modifications. The effects of both metals on most of the measured photosynthesis parameters were similar, but the accumulation of Cd in the cotyledons was significantly higher than Cu accumulation. Thus Cu was more toxic for the photosynthesis of cucumber cotyledons than Cd.     

Overexpression of sweet pepper glycerol-3-phosphate acyltransferase gene enhanced thermotolerance of photosynthetic apparatus in transgenic tobacco

In order to investigate the relationship between the lipid composition in thylakoid membrane and thermostability of pho-tosynthetic apparatus, tobacco transformed with sweet pepper sense glycerol-3-phosphate acyltransferase (GPA T) gene were used to analyze the lipid composition in thylakoid membrane, the net photosynthetic rate and chlorophyll fluorescence parameters under high temperature stress. The results showed that the saturated extent of monogalactosyldiacylglycerol (MGDG), suifoquinovosyldiacylglycerol, digalactosyldiacylglycerol and phosphatidylglycerol in thylakoid membrane of transgenic tobacco T1 lines increased generally. Particularly, the saturated extent in MGDG increased obviously by 16.2% and 12.0% in T1-2 and T1-1, respectively. With stress temperature elevating, the maximum efficiency of photosystem Ⅱ the two lines and wild type tobacco plants decreased gradually, but those parameters decreased much less in transgenic plants. Even though the recovery process appeared differently in the donor and acceptor side of PSII in transgenic tobacco compared with wild-type plants, the entire capability of PSII recovered faster in transgenic tobacco, which was shown in Increase in saturated extent of thylakoid membrane Iipids in transgenic plants enhanced the stability of photosynthetic apparatus under high temperature stress.     

Thermal dissipation during photosynthetic induction and subsequent dark recovery as measured by photoacoustic signals

The thermal photoacoustic signal (279 Hz) and the chlorophyll (Chl) fluorescence of radish cotyledons (Raphanus sativus L.) were measured simultaneously. The signals were recorded during a photosynthetic induction with actinic radiation of different quantum fluence rates [20, 200, and 1200 μmol(PAR-quantum) m^-2 s^-1]. The rise of these signals upon irradiation saturating photosynthesis was followed in the steady state of the induction and during the subsequent dark-recovery (i.e., in dark periods of 1, 5, 15, and 45 min after the induction). From these values various parameters (e.g., quantum yield, photochemical loss, different types of quenching coefficients) were calculated. The results show that heat dissipation detected by photoacoustic measurements is neither low, constant, nor always parallel to Chl fluorescence. Therefore, the thermal signal should always be measured in order to fully understand the way leaves convert energy taken up by PAR absorption. This helps in the interpretation of photosynthesis under different natural and anthropogenic conditions (stress and damage effects). This revised version was published online in September 2006 with corrections to the Cover Date.     

Impacts of Elevated CO2 Concentration on Biochemical Composition, Carbonic Anhydrase, and Nitrate Reductase Activity of Freshwater Green Algae

To investigate the biochemical response of freshwater green algae to elevated CO2 concentrations, Chlorella pyrenoidosa Chick and Chlamydomonas reinhardtii Dang cells were cultured at different CO2 concentrations within the range 3-186μmol/L and the biochemical composition, carbonic anhydrase (CA),and nitrate reductase activities of the cells were investigated. Chlorophylls (Chl), carotenoids, carbonhydrate,and protein contents were enhanced to varying extents with increasing CO2 concentration from 3-186μmol/L. The CO2 enrichment significantly increased the Chl a/Chl b ratio in ChloreUa pyrenoidosa, but not in Chlamydomonas reinhardtii. The CO2 concentration had significant effects on CA and nitrate reductase activity. Elevating CO2 concentration to 186μmol/L caused a decline in intracellular and extracellullar CA activity. Nitrate reductase activity, under either light or dark conditions, in C. reinhardtii and C. pyrenoidosa was also significantly decreased with CO2 enrichment. From this study, it can be concluded that CO2 enrichment can affect biochemical composition, CA, and nitrate reductase activity, and that the biochemical response was species dependent.     

Physiological and Biochemical Changes in Microcystis aeruginosa Qutz. in Phosphorus Limitation

Effects of phosphorus limitation on the physiological and biochemical changes of the freshwater bloom alga Microcystis aeruginosa Qutz. are reported in the present study. As a result of phosphorus limitation, biomass was controlled to some extent and the protein content per cell in vivo decreased. However,the carbohydrate content per cell was higher in phosphorus limitation over the 8d of cultivation. Soluble proteins were distinct in the media, whereas phosphorus deficiency induced the presence of a unique protein (16.2 kDa). Under conditions of phosphorus limitation, the activities of both superoxide dismutase and peroxidase per cell in vivo were lower than under normal conditions in the last cultivation. The in vivo absorption spectra of cells showed chlorophyll absorption peaks at 676 and 436nm, over 10nm red-shifted from the normal position; cells showed an absence of a chlorophyll c with an in viva absorption peak at 623nm and an extraction absorption peak at 617nm. The chlorophyll a/carotene and chlorophyll a/xanthophylls ratios decreased under conditions of phosphorus limitation, photosynthetic efficiency (Fv/Fm) was clearly lower, and the low-temperature fluorescence emission spectra indicated a higher peak at 683nm and a lower peak at 721nm relatively, with the 721nm peak drifting slightly to the red and the 683 nm peak strengthened with a weakened 692nm shoulder peak.     

Arabidopsis tic62 trol Mutant Lacking Thylakoid- Bound Ferredoxin-NADP＋ Oxidoreductase Shows Distinct Metabolic Phenotype

Ferredoxin-NADP＋ oxidoreductase （FNR）, functioning in the last step of the photosynthetic electron transfer chain, exists both as a soluble protein in the chloroplast stroma and tightly attached to chloroplast membranes. Surface plasmon resonance assays showed that the two FNR isoforms, LFNR1 and LFNR2, are bound to the thylakoid membrane via the C-terminal domains of Tic62 and TROL proteins in a pH-dependent manner. The tic62 trol double mutants contained a reduced level of FNR, exclusively found in the soluble stroma. Although the mutant plants showed no visual phenotype or defects in the function of photosystems under any conditions studied, a low ratio of NADPH/NADP~ was detected. Since the CO2 fixation capacity did not differ between the tic62 trol plants and wild-type, it seems that the plants are able to funnel reducing power to most crucial reactions to ensure survival and fitness of the plants. However, the activity of malate dehydrogenase was down-regulated in the mutant plants. Apparently, the plastid metabolism is able to cope with substantial changes in directing the electrons from the light reactions to stromal metabolism and thus only few differences are visible in steady-state metabolite pool sizes of the tic62 trol plants.     

Effects of different excitation and detection spectral regions on room temperature chlorophyll <Emphasis Type="Italic">a</Emphasis> fluorescence parameters

The effects of different spectral region of excitation and detection of chlorophyll (Chl) a fluorescence at room temperature on the estimation of excitation energy utilization within photosystem (PS) 2 were studied in wild-type barley (Hordeum vulgare L. cv. Bonus) and its Chl b-less mutant chlorina f2 grown under low and high irradiances [100 and 1 000 μmol(photon) m⁻² s⁻¹]. Three measuring spectral regimes were applied using a PAM 101 fluorometer: (1) excitation in the red region (maximum at the wavelength of 649 nm) and detection in the far-red region beyond 710 nm, (2) excitation in the blue region (maximum at the wavelength of 461 nm) and detection beyond 710 nm, and (3) excitation in the blue region and detection in the red region (660– 710 nm). Non-photochemical quenching of maximal (NPQ) and minimal fluorescence (SV₀), determined by detecting Chl a fluorescence beyond 710 nm, were significantly higher for blue excitation as compared to red excitation. We suggest that this results from higher non-radiative dissipation of absorbed excitation energy within light-harvesting complexes of PS2 (LHC2) due to preferential excitation of LHC2 by blue radiation and from the lower contribution of PS1 emission to the detected fluorescence in the case of blue excitation. Detection of Chl a fluorescence originating preferentially from PS2 (i.e. in the range of 660–710 nm) led to pronounced increase of NPQ, SV₀, and the PS2 photochemical efficiencies (F_V/F_M and F_V′/F_M′), indicating considerable underestimation of these parameters using the standard set-up of PAM 101. Hence PS1 contribution to the minimal fluorescence level in the irradiance-adapted state may reach up to about 80 %.     

Clonal differences in the response of dark and light reactions of photosynthesis to elevated atmospheric CO2 in poplar

Two hybrid poplar (Populus) clones (i.e., fast growing clone Beauprè and slow growing clone Robusta) were grown for two years from cuttings at close spacings in open top chambers (OTCs) under ambient (AC) and elevated [EC = AC + 350 μmol(CO2) mol-1] CO2 treatments. For clone Beauprè no down-regulation of photosynthesis was observed. Two years of growing under EC resulted in an increase in quantum yield of photosystem 2 (PS2), steady state irradiance saturated rate of net photosynthesis (P _Nmax), chlorophyll (Chl) content, and ribulose-1,5-bisphosphate carboxylase/oxygenase (RuBPC) activity for this clone. We suppose that under non-limiting conditions of nitrogen and phosphorus content the response to EC was by building up light-harvesting complexes of PS2 and increasing photochemical efficiency of PS2. Due to a high rate of the primary reactions of photosynthesis and a high RuBPCO activity the end product of the response to EC was an increase in PNmax and a larger saccharides content. The Robusta clone showed a depression in the primary reactions of photosynthesis under EC. We found a decrease in quantum yield of PS2, Chl and phosphorus contents, and in RuBPCO activity. However, an increase in PNmax, saccharides content and Chl a/b ratio was observed. We speculate (1) that the phosphorus deficiency in combination with an increase in CO2 concentrations may lead to a potential damage of the assimilation apparatus of the primary reactions of photosynthesis and to a decrease in photochemical efficiency of PS2; (2) that the primary target of "down-regulation" takes place at PS2 for irradiances above 150 μmol m-2 s-1. This revised version was published online in September 2006 with corrections to the Cover Date.     

Effects of nitrogen addition on photosynthetic characteristics of Leymus chinensis in the temperate grassland of Nei Mongol,China北大核心CSCD

Aims The increased atmospheric nitrogen (N) deposition due to human activity and climate change greatly causes grassland ecosystems shifting from being naturally N-limited to N-eutrophic or N-saturated, and further affecting the growth of grass species. The aims of this study are: 1) to evaluate the effects of different N addition levels on morphology and photosynthetic characteristics of Leymus chinensis; 2) to determine the critical N level to facilitate L. chinensis growth. Methods We conducted a different N addition levels experiment in dominant species in the temperate steppe of Nei Mongol. The aboveground biomass, morphological and leaf physiological traits, pigment contents, chlorophyll a fluorescence parameters and biochemical parameters of L. chinensis were investigated. Important findings Our results showed that aboveground biomass first increased and then decreased with the increased N, having the highest values at the 10 g N·m-2·a?1 treatment, but the 25 g N·m-2·a?1 still significantly increased the aboveground biomass relative to 0 g N·m-2·a?1. Leymus chinensis accommodate low N situation through allocating less N to carboxylation system and decreasing leaf mass per area (LMA) in order to get more light energy. Moderate N addition captured more light energy through increasing total chlorophyll (Chl) contents and decreasing the ratio of Chl a/b. Moderate N addition increased LMA, carboxylation efficiency, maximum car boxylation rate (Vcmax), maximum electron transport rate (Jmax) and decreased Jmax/Vcmax, thus allocating more N to carboxylation system to enhance carboxylation capability. Moreover, the photochemical activity of PSII was increased through higher effective quantum yield of PSII photochemistry, electron transport rate and photochemical quenching coefficient. Excessive N addition had negative effects on physiological variables of L. chinensis due to lower carboxylation capability and photochemical activity of PSII, further leading to decreased net photosynthetic rate, whereas increased non-photochemical quenching coefficient and carotenoids played the role in the dissipation of excess excitation energy. Overall, moderate N addition facilitated the photosynthetic characteristics of dominant species, but excessive N addition inhibited photosynthetic characteristics. The most appropriate N addition for the growth of L. chinensis was 5-10 g N·m-2·a?1 in the temperate steppe of Nei Mongol, China.     

Substitutions of the Conserved Gly47 Affect the CF1 Inhibitor and Proton Gate Functions of the Chloroplast ATP Synthase ε Subunit

The conserved residue Gly47 of the chloroplast ATP synthase ε subunit was substituted with Leu, Arg, Ala and Glu by site-directed mutagenesis. This process generated the mutants εG47L, εG47R, εG47A and εG47E, respectively. All the ε variants showed lower inhibitory effects on the soluble CF1(-ε) Ca^2 -ATPase compared with wild-type ε. In reduced conditions, εG47E and εG47R had a lower inhibitory effect on the oxidized CF1(-ε) Ca^2 -ATPase compared with wild-type ε. In contrast, εG47L and εG47Aincreased the Ca^2 -ATPase activity of soluble oxidized CF1(-ε). The replacement of Gly47 significantly impaired the interaction between the subunit ε and γ in an in vitro binding assay. Further study showed that all ε variants were more effective in blocking proton leakage from the thylakoid membranes. This enhanced ATP synthesis of the chloroplast and restored ATP synthesis activity of the reconstituted membranes to a level that was more efficient than that achieved by wild-type ε. These results indicate that the conserved Gly47 residue of the ε subunit is very important for maintaining the structure and function of the ε subunitand may affect the interaction between the ε subunit, β subunit of CF1 and subunit Ⅲ of CF0, therebyregulating the ATP hydrolysis and synthesis, as well as the proton translocation role of the subunit Ⅲ of CF0.     

Gradual Disassembly of Photosystem 2 In Vivo Induced by Excess Irradiance. A Hypothesis Based on Changes in 77 K Fluorescence Spectra of Chlorophyll a in Barley Leaves

Effects of short-term exposure to different irradiances on the function of photosystem 2 (PS2) were studied for barley grown at low (LI; 50 μmol m⁻² s⁻¹) and high (HI; 1 100 μmol m⁻² s⁻¹) irradiances. HI barley revealed higher ability to down-regulate the light-harvesting within PS2 after exposure to high irradiance as compared to LI plants. This ability was estimated from the light-induced decreases of F685/F742 and E476/E436 in emission and excitation spectra of 77 K chlorophyll (Chl) a fluorescence in vivo which was 65 and 10 % for HI plants as compared to 30 and 2 % for LI plants, respectively. For LI plants this protective down-regulation of the light-harvesting of PS2 was saturated at 430 μmol m⁻² s⁻¹, and progressive PS2 photodamage was induced at higher irradiances. After exposure of LI segments to 2 200 μmol m⁻² s⁻¹ a pronounced maximum at 700 nm appeared in emission spectrum of 77 K Chl a fluorescence. Based on complementary analysis of 77 K excitation spectra measured at the emission wavelength 685 nm we suggest that this emission maximum may be attributed to the formation of aggregates of light-harvesting complexes of PS2 (LHC2) with part of PS2 core during progressive PS2 photodamage. Our results can be explained assuming different contributions of LHC2 and PS2 core to the total nonradiative dissipation of absorbed excitation energy for the LI and HI barley. This revised version was published online in August 2006 with corrections to the Cover Date.     

Chlorophyll Fluorescence Detected Passively by Difference Reflectance Spectra of Wheat （Triticum aestivum L.） Leaf

The chlorophyll fluorescence （CF） signature emitted from vegetation provides an abundance of information regarding photosynthetics activity and has been used as a powerful tool to obtain physiological information of plant leaves in a non-invasive manner. CF is difficult to quantify because the CF signal is obscured by reflected light. In the present study, the apparent reflectance spectra of wheat （Triticum aestivum L.） leaves were measured under illuminations with and without filtering by three specially designed long-wave pass edge filters; the cut-off wavelengths of the three filters were 653.8, 678.2, and 694. l nm at 50% of maximum transmittance. The CF spectra could be derived as the reflectance difference spectra of the leaves under illuminations with and without the long wave pass edge filters. The ratio of the reflectance difference at 685 and 740 nm （Dif685/Dif740） was linear correlated with the CF parameters （maximal photochemical efficiency Fv/Fm, and the yield of quantum efficiency） measured by the modulated fluorometer. In addition, the ratio reflected the water stress status of the wheat leaf, which was very high when water deficiency was serious. This method provides a new approach for detecting CF and the physiological state of crops.