High irradiance and water stress induce alterations in pigment composition and chloroplast activities of primary wheat leaves

Remarkable changes were observed in chlorophyll (Chl) (a+b), carotenoids (Car), and protein content of leaves and fluorescence emission, polarisation, excitation energy transfer, lipid peroxidation and DCPIP photoreduction activity in isolated chloroplasts of wheat leaves grown under moderate irradiance (MI, 15 W m⁻², control) and subsequently exposed to high irradiance stress (HIS, 250 W m⁻²), water stress (WS, 5 % aqueous polyethylene glycol-4000 solution) and HIS+WS simultaneously, during mature and senescence phase. In the stress exposed samples the Chl, Car and protein contents and kinetics of Hill activity significantly declined. Decrease in excitation energy transfer and increase in membrane polarisation and accumulation of malondialdehyde (MDA) in chloroplasts were also observed. The effect was more pronounced when the seedlings were treated with HIS+WS simultaneously. These observations suggest additive and a possible synergetic action of HIS and WS causing faster loss of pigments and protein content, intense changes in membrane properties including photochemical function, compared to samples exposed to either of the stresses individually.     

High irradiance-induced changes in carotenoid composition and increase in non-photochemical quenching of Chl a fluorescence in primary wheat leaves

The effect of acclimation to high irradiance stress (HIS, 250 Wm-2) in wheat leaves grown under three different irradiances was investigated by HPLC analyses of pigments, chlorophyll a fluorescence parameters and photochemical activities of chloroplasts. Significant loss of beta-carotene was observed compared to the xanthophylls in all three types of seedlings exposed to HIS. However, the effect of HIS on neoxanthin and lutein contents was not significant. The loss of partial electron transport (Asc-DCPIP to MV, PSI activity) was less than the whole chain (H2O to MV) and PS II activity (H2O to DCPIP) suggesting that PS I is less susceptible to HIS compared to PS II. The percent of reductions in Fv/Fm and phi PS II were less in plants grown under high irradiance (HI-1, 30 Wm-2 and HI-2, 45 Wm-2) compared to those grown under moderate irradiance (MI, 15 Wm-2). On the other hand, the percent of NPQ increased more in the leaves of HI plants compared to the leaves of MI when exposed to HIS which suggests a more efficient non-radiative dissipation of excess excitation energy in HI plants compared to MI. These observations suggest that plants grown under relatively high irradiance are better adapted to HIS condition.     

Photosynthetic Characteristics of Chloroplasts of Primary Wheat Leaves Grown under Different Irradiance

The rate of accumulation of total chlorophyll (Chl) and carotenoids (Car) of leaves grown under high irradiance, HI (30 and 45 W m^–2) was faster than at moderate irradiance, MI (15 W m^–2). However, the senescence phase started earlier in the samples and proceeded at a faster rate. Chl a/b and Chl (a+b)/Car values showed faster loss of Chl a (compared to Chl b) and Chl (a+b) (compared to Car) in HI leaves. Protein accumulation and loss were also similar to that of Chl (a+b) content. Increase in Chl fluorescence during the development phase may suggest a gradual change in thylakoid organisation, however, the temporal kinetics were different in HI and MI samples. Increase in fluorescence polarisation during senescence of HI leaves compared to the control (MI) suggests conversion of thylakoid membranes to gel phase. Chloroplasts prepared from HI seedlings showed higher rate of photochemical activities, however, the activity declined earlier and at faster rate compared to the control.     

The Photosynthesis and Chlorophyll a Fluorescence in Seedlings of Kandelia Candel (L.) Druce Grown under Different Nitrogen and NaCl Controls

Kandelia candel (L.) Druce is the dominant mangrove species on the west coast of northern Taiwan. We have measured the net photosynthetic rate (P _N) and chlorophyll (Chl) a fluorescence of seedlings grown at combinations of two nitrogen (0.01 and 0.1 mM) and two NaCl (250 and 430 mM NaCl) controls. With the same nitrogen level, seedlings grown at higher salinity (HS) had a significantly lower P _N and stomatal conductance (g _s) than those at lower salinity (LS). An increase in nitrogen availability significantly elevated P _N and g _s of the LS-grown seedlings. Compared to dark adapted leaves, the maximum quantum yield of photosystem 2 (PS2) (F_v/F_m) of leaves exposed to PFDs of 1200 and 1600 μmol m^-2 s^-1 for 2 h was significantly reduced. The degree of F_v/F_m reduction differed among leaves of the four types of treated plants. Chl fluorescence quenching analysis revealed differences among the examined plants in coefficients of non-photochemical and photochemical quenching. This revised version was published online in June 2006 with corrections to the Cover Date.     

Susceptibility of green leaves and green flower petals of CAM orchid <Emphasis Type="Italic">Dendrobium</Emphasis> cv. Burana Jade to high irradiance under natural tropical conditions

Photosynthetic rates of green leaves (GL) and green flower petals (GFP) of the CAM plant Dendrobium cv. Burana Jade and their sensitivities to different growth irradiances were studied in shade-grown plants over a period of 4 weeks. Maximal photosynthetic O₂ evolution rates and CAM acidities [dawn/dusk fluctuations in titratable acidity] were higher in leaves exposed to intermediate sunlight [a maximal photosynthetic photon flux density (PPFD) of 500–600 μmol m⁻² s⁻¹] than in leaves grown under full sunlight (a maximal PPFD of 1 000–1 200 μmol m⁻² s⁻¹) and shade (a maximal PPFD of 200–250 μmol m⁻² s⁻¹). However, these two parameters of GFP were highest in plants grown under the shade and lowest in full sun-grown plants. Both GL and GFP of plants exposed to full sunlight had lower predawn F_v/F_m [dark adapted ratio of variable to maximal fluorescence (the maximal photosystem 2 yield without actinic irradiation)] than those of shade-grown plants. When exposed to intermediate sunlight, however, there were no significant changes in predawn F_v/F_m in GL whereas a significant decrease in predawn F_v/F_m was found in GFP of the same plant. GFP exposed to full sunlight exhibited a greater decrease in predawn F_v/F_m compared to those exposed to intermediate sunlight. The patterns of changes in total chlorophyll (Chl) content of GL and GFP were similar to those of F_v/F_m. Although midday F_v/F_m fluctuated with prevailing irradiance, changes of midday F_v/F_m after exposure to different growth irradiances were similar to those of predawn F_v/F_m in both GL and GFP. The decreases in predawn and midday F_v/F_m were much more pronounced in GFP than in GL under full sunlight, indicating greater sensitivity in GFP to high irradiance (HI). In the laboratory, electron transport rate and photochemical and non-photochemical quenching of Chl fluorescence were also determined under different irradiances. All results indicated that GFP are more susceptible to HI than GL. Although the GFP of Dendrobium cv. Burana Jade require a lower amount of radiant energy for photosynthesis and this plant is usually grown in the shade, is not necessarily a shade plant.     

Structure-function correlation during the etioplast-chloroplast transition in cucumber cotyledonary leaves

We studied the development of chloroplasts from etioplasts in the cotyledonary leaves of 4-d-old dark-grown cucumber (Cucumis sativus) seedlings after irradiation (20 μmol m^-2 s^-1). Upon irradiation, the triggering of chlorophyll (Chl) synthesis and accumulation showed a relatively short lag phase. The irradiation of etiolated seedlings initiated the synthesis of apoproteins of pigment-protein complexes. While Chl-protein 2 (CP2) was detected at 6 h after irradiation, CP1 only after 29 h. The appearance and accumulation of some of the apoproteins were monitored by Western-blotting. LHC2 apoprotein was detected after a 6 h-irradiation. The amounts of D1 protein of photosystem (PS) 2 and PsaA/B protein of PS1 were quantitated by ELISA. Further, the thylakoid membrane function during this time period in terms of PS1- and PS2-mediated electron transfer activity and intersystem electron pool size were analyzed. While PS1 activity was detected after 4 h, PS2-mediated O₂ evolution was detected only after a 17 h-irradiation. F_v/F_m value of Chl a fluorescence measurements indicated that the photochemical efficiency of these leaves reached its maximum after 29 h of irradiation. The intersystem pool size of cotyledonary leaves was equivalent to that of the control cotyledonary leaves grown for 25 h under continuous irradiation. Thus etioplasts develop into fully functional chloroplasts after approximately 25 h when 4 d-dark grown cucumber seedlings are continuously moderately irradiated. The development of photosynthetic electron transport chain seems to be limited in time at the level of PS2, possibly at the donor side. This revised version was published online in September 2006 with corrections to the Cover Date.     

Functional responses of Acer species to two simulated forest gap environments: leaf-level properties and photosynthesis

Seedlings of eight forest maple (Acer L.) species were grown outdoors through a full season under two irradiation treatments: (a) “gap edge” with a photosynthetic photon flux density of 30 μmol m-² s-¹ and a red:far-red ratio of 0.55, and (b) “gap centre” with 400 μmol m-² s-¹ and a red:far-red ratio of 1.12. Area-based leaf nitrogen concentration was greater in gap centre-grown seedlings, whereas, except for A. saccharum, area-based chlorophyll (Chl) (a+b) was higher in gap edge-grown plants. There was also a significantly lower Chl a/b ratio in gap edge-grown plants. Maximum photosynthetic rate (P _max ) was 60 % higher in the gap-centre treatment. These results are consistent with the functional expectation that shade-acclimated plants will increase their radiant-energy harvesting capacity as a result of limited photon input while gap-acclimated plants will operate more efficiently under bright irradiance by increasing their carboxylation capacity. This inverse relationship between the capacity of the light-harvesting component and the carboxylation component is, however, only partially supported by Chl fluorescence measurements of intact leaves. Compared to gap centre-grown plants, the lower total fluorescence quenching in gap edge-grown plants indicated a lower carboxylation capacity that was in accord with the observed P _max . However, edge-grown seedlings did not show the expected improvement in light-harvesting efficiency and reduction in electron transport of photosystem 2 inferred from their marginally greater t_1/2 and lower F_v/F_m, respectively. Hence while maples acclimated to different irradiation levels by adjusting leaf N and Chl contents, they showed limited acclimation potential at the photosystem level. Variations in the leaf traits examined had only minor effect on low irradiance photosynthesis and sunfleck utilization. This revised version was published online in August 2006 with corrections to the Cover Date.     

Size and Power of Chlorophyll a Molecule

On the basis of literature and my calculations it was established that a chlorophyll (Chl) particle anchored with a phytol chain to the thylakoid membrane takes up about 1 nm² of the surface area. At an irradiance of 287 W m^-2 the leaves of cabbage seedlings become saturated with photosynthetically active radiation (PAR) thus reaching the maximum photosynthetic rate of 100 µg(C) m^-2 s^-1, that is 5 CO₂ molecules per 1 nm² per second, and the maximum power with which the Chl particle supplies the process of photosynthesis is 15 aJ.     

Effects of Nitrogen Deficiency on Gas Exchange,Chlorophyll Fluorescence,and Antioxidant Enzymes in Leaves of Rice Plants

Gas exchange, chlorophyll (Chl) fluorescence, and contents of photosynthetic pigments, soluble proteins (ribulose-1,5-bisphosphate carboxylase/oxygenase, RuBPCO), and antioxidant enzymes were characterized in the fully expanded 6^th leaves in rice seedlings grown on either complete (CK) or on nitrogen-deficient nutrient (N-deficiency) solutions during a 20-chase period. Compared with the control plants, the lower photosynthetic capacity at saturation irradiance (P _max) was accompanied by an increase in intercellular CO₂ concentration (C_i), indicating that in N-deficient plants the decline in P _max was not due to stomatal limitation but due to the reduced carboxylation efficiency. The fluorescence parameters _PS2, F_v/F_m, electron transport rate (ETR), and q_P showed the same tendency as P _max in N-deficient plants. Correspondingly, a higher q_N paralleled the rise of the ratio of carotenoid (Car) to Chl contents. However, F_v/F_m was still diminished, suggesting that photoinhibition did occur in the photosystem 2 (PS2) reaction centres. In addition, the activities of antioxidant enzymes on a fresh mass basis were gradually lowered, leading to the aggravation of membrane lipid peroxidation with the proceeding N-deficiency. The accumulation of malonyldialdehyde resulted in the lessening of Chl and soluble protein content. Analyses of regression showed PS2 excitation pressure (1 - q_P) was linearly correlated with the content of Chl and inversely with soluble protein (particularly RuBPCO) content. There was a lag phase in the increase of PS2 excitation pressure compared to the decrease of RuBPCO content. Therefore, the increased excitation pressure under N-deficiency is probably the result of saturation of the electron transport chain due to the limitation of the use of reductants by the Calvin cycle. Rice plants responded to N-deficiency and high irradiance by decreasing light-harvesting capacity and by increasing thermal dissipation of absorbed energy.     

Diurnal changes in chlorophyll fluorescence and light utilization in Colocasia esculenta leaves grown in marshy waterlogged area

Diurnal cycle of chlorophyll fluorescence parameters was done in Colocasia esculenta L. (swamp taro) grown in marshy land under sun or under shade. The sun leaves maintained higher electron transport rate (ETR) and steady state to initial fluorescence ratio (F_s/F₀) than shade leaves. In spite of lower ETR, higher photochemical quenching (PQ), and effective quantum yield of photosystem 2 (Φ_PS2) was evident in shade plants compared to plants exposed to higher irradiance. ETR increased linearly with increase in irradiance more under low irradiance (r ² = 0.84) compared to higher irradiance (r ² = 0.62). The maximum quantum yield of PS 2 (F_v/F_m) did not differ much in sun and shade leaves with the exception of midday when excess of light energy absorbed by plants under sun was thermally dissipated. Hence swamp taro plants adopted different strategies to utilize radiation under different irradiances. At higher irradiance, there was faster decline in proportion of open PS 2 centers (PQ) and excess light energy was dissipated through non-photochemical quenching (NPQ). Under shade, absorbed energy was effectively utilized resulting in higher Φ_PS2.     

Changes in photosynthetic apparatus during dark incubation of detached leaves from control and ultraviolet-B treatedVigna seedlings

Changes in various components of photosynthetic apparatus during the 4 d dark incubation at 25°C of detached control and ultraviolet-B (UV-B) treatedVigna unguiculata L. leaves were examined. The photosynthetic apparatus was more degraded in younger control seedlings and for a longer time UV-B treated seedlings than in the older or for a shorter time UV-B treated seedlings. This was shown by determining the losses in chlorophyll (Chl) and protein contents, variable fluorescence yield, photosystem (PS) 2, PS1 and ribulose-1,5-bisphosphate carboxylase (RuBPC) activities, and photosynthetic¹⁴CO₂ fixation. In contrast, the Car/Chl ratio increased during the dark incubation due to less expressed degradation of Car.     

Acclimation of shade-tolerant and light-resistant <Emphasis Type="Italic">Tradescantia</Emphasis> species to growth light: chlorophyll <Emphasis Type="Italic">a</Emphasis> fluorescence,electron transport,and xanthophyll content

In this study, we have compared the photosynthetic characteristics of two contrasting species of Tradescantia plants, T. fluminensis (shade-tolerant species), and T. sillamontana (light-resistant species), grown under the low light (LL, 50–125 µmol photons m^?2 s^?1) or high light (HL, 875–1000 µmol photons m^?2 s^?1) conditions during their entire growth period. For monitoring the functional state of photosynthetic apparatus (PSA), we measured chlorophyll (Chl) a emission fluorescence spectra and kinetics of light-induced changes in the heights of fluorescence peaks at 685 and 740 nm (F ₆₈₅ and F ₇₄₀). We also compared the light-induced oxidation of P₇₀₀ and assayed the composition of carotenoids in Tradescantia leaves grown under the LL and HL conditions. The analyses of slow induction of Chl a fluorescence (SIF) uncovered different traits in the LL- and HL-grown plants of ecologically contrasting Tradescantia species, which may have potential ecophysiological significance with respect to their tolerance to HL stress. The fluorometry and EPR studies of induction events in chloroplasts in situ demonstrated that acclimation of both Tradescantia species to HL conditions promoted faster responses of their PSA as compared to LL-grown plants. Acclimation of both species to HL also caused marked changes in the leaf anatomy and carotenoid composition (an increase in Violaxanthin?+?Antheraxantin?+?Zeaxanthin and Lutein pools), suggesting enhanced photoprotective capacity of the carotenoids in the plants grown in nature under high irradiance. Collectively, the results of the present work suggest that the mechanisms of long-term PSA photoprotection in Tradescantia are based predominantly on the light-induced remodeling of pigment-protein complexes in chloroplasts.     

Strong light elevates thermotolerance of photosynthetic apparatus and the content of membranes and polar lipids in wheat leaves

The influence of excess irradiance on resistance of wheat (Triticum aestivum L.) photosynthetic apparatus to heating in darkness and in the light was investigated and compared with changes in leaf cell ultra-structure and composition of cell lipids and fatty acids. The leaves of 14- to 16-day-old plants grown at low irradiance (about 20 W/m²) were exposed for 1 h to irradiance of 370 or 600 W/m² PAR. Using infrared gas analysis, we found that the preexposure of leaves to excess irradiation elevated resistance of apparent photosynthesis to 10-min heat treatment at 40–45°C. The rate of Hill reaction (reduction of 2,6-dichlorophenolindophenol by isolated chloroplasts) was higher for leaves heated at high irradiance than for leaves heated in darkness. During illumination of leaves with strong light, mesophyll cells became more abundant in mitochondria and peroxysomes, as well as in cisternae of endoplasmic reticulum and Golgi complex. The chloroplast thylakoids and grana became more extensive and numerous. At the same time, the leaf content of main classes of membrane glycerolipids increased in parallel with the increase in the phospholipid/glycolipid and lipid/chlorophyll ratios. The unsaturation index of fatty acids of membrane lipids increased because of the elevated content of linolenic acid. Thus, excessive light (not fully utilized in photosynthesis) induced in wheat leaves a series of nonspecific adaptive changes that were similar to those occurring under the action of other environmental factors, such as heat shock, cooling, salinity, and osmotic stresses.     

Changes in photosynthetic apparatus during dark incubation of detached leaves from control and ultraviolet-B treatedVigna seedlings

Changes in various components of photosynthetic apparatus during the 4 d dark incubation at 25°C of detached control and ultraviolet-B (UV-B) treatedVigna unguiculata L. leaves were examined. The photosynthetic apparatus was more degraded in younger control seedlings and for a longer time UV-B treated seedlings than in the older or for a shorter time UV-B treated seedlings. This was shown by determining the losses in chlorophyll (Chl) and protein contents, variable fluorescence yield, photosystem (PS) 2, PS1 and ribulose-1,5-bisphosphate carboxylase (RuBPC) activities, and photosynthetic¹⁴CO₂ fixation. In contrast, the Car/Chl ratio increased during the dark incubation due to less expressed degradation of Car.     

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

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

The kinetics of P515 in relation to the lipid composition of the thylakoid membrane

Flash-induced P515 absorbance changes have been studied in dark-adapted chloroplasts isolated from spinach plants grown under two different light intensities. The slow component (reaction 2), normally present in the P515 response of chloroplasts isolated from plants grown at an intensity of 60 W · m^–2, was largely reduced in chloroplasts isolated from plants grown at an intensity of 6 W · m^–2. This reduction of the slow component in the P515 response appeared to be coincident with an alteration in the lipid composition of the thylakoid membrane. Mainly the ratio monogalactosyldiacylglycerol to digalactosyldiacylglycerol appeared to be altered. In thylakoids from plants grown at 6 W · m^–2, the ratio was approximately 35% lower than that of plants grown at 60 W · m^–2. The amount of both cytochromeb ₅₆₃ and cytochromef was largely reduced in chloroplasts isolated from plants grown at low light intensity. These results may indicate a possible correlation between structural organization of the thylakoid membrane and the kinetics of the flash-induced P515 response.     

Cultivation of the brown alga <Emphasis Type="Italic">Hizikia fusiformis</Emphasis> (Harvey) Okamura: stress resistance of artificially raised young seedlings revealed by chlorophyll fluorescence measurement

Commercial farming of the intertidal brown alga Hizikia fusiformis (Harvey) Okamura in China and South Korea in the sea depends on three sources of seedlings: holdfast-derived regenerated seedlings, young plants from wild population and zygote-derived seedlings. Like many successfully farmed seaweed species, the sustainable development of Hizikia farming will rely on a stable supply of artificial seedlings via sexual reproduction under controlled conditions. However, the high rate of detachment of seedlings after transfer to open sea is one of the main obstacles, and has limited large-scale application of zygote-derived seedlings. To seek the optimal condition for growing seedlings on substratum in land-based tanks for avoidance of detachment in this investigation, young seedlings were grown in both outdoor tanks exposed directly to sunlight and in indoor raceway tanks in reduced, filtered sunlight. Results showed that young seedlings, immediately after fertilization, could withstand a daily fluctuation of direct solar irradiance up to a level of 1800 μmol photons m⁻² s⁻¹, and maintained a faster growth rate than seedlings grown in indoor tanks. Detailed experiments by use of chlorophyll fluorescence measurements further demonstrated that the overnight (12 h) recovery of optimal fluorescence quantum yield (F_v/F_m) of seedlings after 1 h treatment at 40°C was 98%, and the 48 h recovery of F_v/F_m of seedlings after 1 h exposure to 1800 μmol m⁻² s⁻¹ was 92%. Forty-one-day-old seedlings showed no significant decrease of optimal fluorescence quantum yield at salinity ranging from 30 to 5 ppt for a treatment up to 17 h. Six-hour desiccation treatment did not have any influence on the optimal fluorescence quantum yield. Exposure to 18 mmol L⁻¹ sodium hypochlorite for 10 min did not damage the PSII efficiency, and thus could be used to remove epiphytic algae. The strong tolerance of young seedlings to high temperature, high irradiance, low salinity and desiccation found in this investigation supports the view that mass production of Hizikia seedlings should be performed in ambient light and temperature instead of in shaded greenhouse tanks.     

Response of senescing rice leaves to flooding stress

Flooding stress (FS) induced changes in pigment and protein contents and in photochemical efficiency of thylakoid membranes of chloroplasts were investigated during senescence of primary leaves of rice seedlings. Leaf senescence was accompanied by loss in 2,6-dichlorophenolindophenol (DCPIP) photoreduction, rate of oxygen evolution, quantum yield of photosystem 2 with an increase in MDA accumulation, and non-photochemical quenching (NPQ) of chlorophyll fluorescence. These changes were further aggravated when the leaves during this period experienced FS. The increase in NPQ value under stress may indicate photosynthetic adaptation to FS.     

Effect of Pb ions on superoxide dismutase and catalase activities in leaves of pea plants grown in high and low irradiance

The role of irradiance on the activity of antioxidant enzymes: superoxide dismutase (SOD) and catalase (CAT) was examined in the leaves of Pisum sativum L. plants grown under low (LL) or high (HL) irradiance (PPFD 50 or 600 μmol m⁻² s⁻¹) and exposed after detachment to 5 mM Pb (NO₃)₂ for 24 h. The activities of both enzymes increased in response to LL compared with HL and no effect of Pb ions was observed. Photosystem (PS) 1 and PS 2 activities were also investigated in chloroplasts isolated from these leaves. LL lowered PS 1 electron transport rate and changes in photochemical activity of PS 1 induced by Pb²⁺ were visible only in the chloroplasts isolated from leaves of LL grown plants. PS 2 activity was influenced similarly by Pb ions at both PPFD. This study demonstrates that leaves of HL grown plants were less sensitive to lead toxicity than those from LL grown plants. Changes in electron transport rates were the main factors responsible for the generation of reactive oxygen species in the chloroplasts and as a consequence, in induction of antioxidant enzymes.     

The influence of ultraviolet-B radiation on the growth, pigment production and chlorophyll fluorescence of Norway spruce seedlings

Norway spruce (Picea abies (L.)Karst.) from seven seed sources was grown in a greenhouse with 8.3 and 14.7 kJ·m⁻²·d⁻¹ m UV-B_BE (biologically effective UV-B: 280–320 nm) irradiation, and with no supplemental irradiation as control. The seedlings total biomass (dry weight) and shoot growth decreased with high UV-B treatment but spruce from low elevation seed sources were more affected. The seedlings grown at the highest UV-B irradiance (14.7 kJ·m⁻²·d⁻¹) showed from 5 to 38% inhibition of total biomass and 15 to 70 % shoot growth inhibition. Norway spruce populations from higher altitude seed sources manifested greater tolerance to UV-B radiation compared to plants from low altitudes. Changes in phospholipids and protective pigments were also determined. The plants grown at the lower UV-B irradiance (8.3 kJ·m⁻²·d⁻¹) showed greater ability to concentrations UV-B-absorbing pigments then control plants. Chlorophyll a fluorescence parameter R_fd, (R_fd=(F_m-F_s)/F_s) showed a significant decrease in needles of UV-B treated plants and this correlated with the altitude of seed source. Exposure to UV-B affect levels of the ratio of variable to maximum fluorescence (F_v/F_m). Results from this study suggest that the response to increased levels of UV-B radiation is depended upon the ecotypic differentiation of Norway spruce and involved changes in metabolites in plant tissues.