Influence of solar ultraviolet radiation on photosynthesis and motility of marine phytoplankton

Abstract: The effects of solar ultraviolet radiation (UV) on carbon uptake, oxygen evolution and motility of marine phytoplankton were investigated in coastal waters at Kristineberg Marine Research Station on the west coast of Sweden (58° 30'N, 11° 30'E). The mean irradiances at noon above the water surface during the investigation period were: photosynthetic active radiation (PAR, 400–700 nm) 1670 μmol m⁻² s⁻¹; ultraviolet-A radiation (UV-A, 320–400 nm) 35.9 W m⁻² and ultraviolet-B radiation (UV-B, 280–320 nm) 1.7 W m⁻². UV-B radiation was much more attenuated with depth in the water column than were PAR and UV-A radiation. UV-B radiation could not be detected at depths greater than 100–150 cm. Inhibition of carbon uptake by UV-A and UV-B in natural phytoplankton populations was greatest at 50 cm depth and the effects of UV-B were greater than those of UV-A. At depths greater than 50 cm there was almost no effect of ultraviolet radiation on carbon uptake. PAR, UV-A and UV-B decreased oxygen evolution by the dinoflagellate Prorocentrum minimum . Inhibition of oxygen evolution was greater after 4 h than 2 h but it was not possible to distinguish the negative effects of the different light regimes. The motility of P. minimum was not affected by PAR, UV-A and UV-B. The importance of exposure of phytoplankton to different light regimes before being exposed to natural solar radiation is discussed.     

In situ responses to elevated CO2 in tropical forest understorey plants

1. Plants growing in deep shade and high temperature, such as in the understorey of humid tropical forests, have been predicted to be particularly sensitive to rising atmospheric CO₂. We tested this hypothesis in five species whose microhabitat quantum flux density (QFD) was documented as a covariable. After 7 (tree seedlings of Tachigalia versicolor and Beilschmiedia pendula ) and 18 months (shrubs Piper cordulatum and Psychotria limonensis, and grass Pharus latifolius ) of elevated CO₂ treatment ( c. 700 μl litre^–1) under mean QFD of less than 11 μmol m^–2 s^–1, all species produced more biomass (25–76%) under elevated CO₂.
2. Total plant biomass tended to increase with microhabitat QFD (daytime means varying from 5 to 11μmol m^–2 s^–1) but the relative stimulation by elevated CO₂ was higher at low QFD except in Pharus .
3. Non-structural carbohydrate concentrations in leaves increased significantly in Pharus (+ 27%) and Tachigalia (+ 40%).
4. The data support the hypothesis that tropical plants growing near the photosynthetic light compensation point are responsive to elevated CO₂. An improved plant carbon balance in deep shade is likely to influence understorey plant recruitment and competition as atmospheric CO₂ continues to rise.     

Measuring and modelling environmental influences on photosynthetic gas exchange in Sphagnum and Pleurozium

A mechanistic model has been used to examine the environmental regulation of photosynthetic gas exchange in moss. The effects of water content on conductance to CO₂ and on photosynthetic capacity during desiccation were calculated from the carbon isotope discrimination data of Williams & Flanagan (1996 , Oecologia 108, pp. 38–46) and combined with the biochemical model of Farquhar et al. (1980 , Planta 149, pp. 78–90). The model includes a simple light attenuation function that imparts curvature to the light response curve for net assimilation, enabling the use of physiologically realistic values for the biochemical parameters. Measurements of gas exchange for Sphagnum and Pleurozium were made in an old black spruce ecosystem over a growing season in order to assign values to parameters in the model. The calculated maximum rates of carboxylation by Rubisco ( V _max) were 5, 14 and 6 μ mol m^–2 s^–1 for Sphagnum during the spring, summer and autumn seasons of 1996, respectively. The increase in V _max during the summer was consistent with an increased allocation of resources to the photosynthetic apparatus. In contrast, no seasonal variation in V _max was observed in Pleurozium with average values of 7, 5 and 7 μ mol m^–2 s^–1 during the spring, summer and autumn, respectively.     

UV-B response of cucumber seedlings grown under metal halide and high pressure sodium/deluxe lamps

UV-B-sensitive (Poinsett) and -insensitive (Ashley) cultivars of cucumber ( Cucumis sativus L.) were grown in growth chambers at 600 μmol m⁻²s⁻¹ of photosynthetically active radiation provided by metal halide (MH) or high pressure sodium/deluxe (HPS/DX) lamps. Plants were irradiated 15 days from seeding for 6 h per day under 18. 2 kJ m⁻² day⁻¹ of biologically effective UV-B (UV-B_BE) radiation. One of the most pronounced effects of UV-B was a 27 to 78% increase in phenylalanine ammonialyase (PAL) activity. UV-B also increased total polyamines. Catalase and superoxide dismutase varied greatly in their response to UV-B. There were no interactive effects on PAL or catalase activity, or total polyamines. There was a UV × PAR source interaction for superoxide dismutase activity. UV-B increased chlorosis and decreased height, dry weight and leaf area. Stem elongation, biomass production, leaf enlargement and chlorosis were greater under HPS/DX lamps than under MH lamps. Chlorosis was greater in Poinsett than in Ashley and in lower leaves than in upper ones. Aside from chlorosis, there were no interactive effects of UV-B, PAR source or cultivar on any of the growth parameters measured, suggesting that the growth response of cucumber seedlings to UV-B is unaffected by PAR source or cultivar. Similarly, except for SOD activity, the biochemical response to UV-B was also not influenced by PAR source or cultivar.     

Inhibition of hypocotyl elongation by ultraviolet-B radiation in de-etiolating tomato seedlings. I. The photoreceptor

Broad-band UV-B radiation inhibited hypocotyl elongation in etiolated tomato ( Lycopersicon esculentum Mill. cv. Alisa Craig) seedlings. This inhibition could be elicited by < 3 μmol m⁻² s⁻¹ of UV-B radiation provided against a background of white light (> 620 μmol m⁻² s⁻¹ between 320 and 800 nm), and was similar in wild-type and phytochrome-1-deficient aurea mutant seedlings. These observations suggest that the effect of UV-B radiation is not mediated by phytochrome. An activity spectrum obtained by delivering 1 μmol m⁻² s⁻¹ of monochromatic UV radiation against a while light background (63 μmol m⁻² s⁻¹ showed maximum effectiveness around 300 nm, which suggests that DNA or aromatic residues in proteins are not the chromophores mediating UV-B induced inhibition of elongation. Chemicals that affect the normal (photo)chemistry of flavins and possibly pterins (KI, NaN, and phenylacetic acid) largely abolished the inhibitor) effect of broad-hand UV-B radiation when applied to the root zone before irradiation. KI was effective at concentrations < 10⁻⁴ M , which have been shown in vitro to be effective in quenching the triplet excited stales of flavins but not fluorescence from pterine or singlet states of flavins. Elimination of blue light or reduction of UV-A, two sources of flavin excitation, promoted hypocotyl elongation, but did not affect the inhibition of elongation evened by UV-B. Kl applied after UV-B irradiation had no effect on the inhibition response. Taken together these findings suggest that the chromophore of the photoreceptor system invoked in UV-B perception by tomato seedlings during de-etiolation may be a flavin.     

ROLE OF ULTRAVIOLET RADIATION IN MAINTAINING THE THREE-DIMENSIONAL STRUCTURE OF A CYANOBACTERIAL MAT COMMUNITY AND FACILITATING NITROGEN FIXATION

Cyanobacterial mat communities were collected in the mangrove forest bordering the Grand Cul de Sac Marin, Guadeloupe, French West Indies, which supports a community of nitrogen fixing cyanobacterial mats established on the trunk and branches of black mangrove ( Avicennia germinans L.). This study presents results that are focused on the mat community and the physiological and morphological adaptations to UV radiation. The dominant surface species of the mat, Nostoc cf commune Vaucher and Scytonema sp., possessed the UV-shielding pigment scytonemin. Mats grown on medium D agar without nitrogen under photosynthetically active radiation (PAR) only, rapidly became disorganized compared with those exposed to PAR + UV-A (320– 400 nm) + UV-B (280–320 nm) irradiation. Concurrent with disorganization, acetylene reduction activity (ARA = one third of N₂ reduction) was severely reduced, whereas mats irradiated with PAR + UV-A + UV-B maintained high ARA activity. Mats incubated for 27 days under PAR + UV-A + UV-B then exposed to PAR only exhibited a 68% stimulation of ARA, whereas ARA values were 33% inhibited in mats incubated with PAR only and then exposed to PAR + UV-A + UV-B. This favorable equilibrium was facilitated by the mats' three-dimensional structure in which the most UV-resistant species, N. commune , covers the surface with UV-sensitive species below this protective covering. The UV stressor was essential for the maintenance of mat structure and ARA.     

Responses of Antarctic Iridaea cordata (Rhodophyta) tetraspores exposed to ultraviolet radiation

In Antarctica ozone depletion is highest during spring, coinciding with the reproduction of many seaweed species. Propagules are the life-stage of an alga most susceptible to environmental perturbations. Therefore, fertile thalli of Iridaea cordata (Turner) Bory (Rhodophyta) were collected in the eulittoral of King George Island (Antarctica) to examine spore susceptibility to ultraviolet radiation (UVR). In the laboratory, freshly released tetraspores were exposed to photosynthetically active radiation (PAR) (400–700 nm), PAR+UV-A (320–700 nm) or PAR+UV-A+UV-B (280–700 nm). Photosynthetic efficiency was measured during 1–8 h of exposure and after 48 h of recovery. Additionally, mycosporine-like amino acids (MAAs) and DNA damage were determined. Saturating irradiance of photosynthesis of freshly released tetraspores was 57 µmol photons m⁻² s⁻¹. Exposure to increasing fluence of PAR reduced photosynthetic efficiency. UVR further decreased the photosynthetic efficiencies of the tetraspores but spores were able to recover completely after UVR exposure and 2 days post-cultivation under low PAR. DNA damage was minimal and lesions were effectively repaired under photoreactivating light. Concentrations of the MAAs shinorine and palythine were higher in tetraspores treated with UVR than in spores only exposed to PAR. Generally, the tetraspores show a good UV tolerance. This flexible response of the tetraspores of this species to changing radiation conditions enables the alga to grow along a considerable depth gradient from the sublittoral to the eulittoral where they can be exposed to enhanced UVBR under conditions of stratospheric ozone depletion.     

The effect of exposure to enhanced UV-B radiation on the penetration of monochromatic and polychromatic UV-B radiation in leaves of Brassica napus

Using quartz optical fibres, penetration of both monochromatic (310 nm) and polychromatic UV-B (280–320 nm) radiation in leaves of Brassica napus L. (cv. Ceres) was measured. Plants were grown under either visible light (750 μmol m⁻² s⁻¹ photosynthetically active radiation) or with the addition of 8. 9 KJ m⁻² day⁻¹ biologically effective UV-B (UV-B_BE) radiation. Results showed that of the 310 nm radiation that penetreated the leaf, 90% was within the intial one third of the leaf with high attenuation in the leaf epidermis, especially in UV-treated plants. Polychromatic UV-B radiation, relative to incident radiation, showed a relatively uniform spectral distribution within the leaf, except for collimated radiation. Over 30% of the UV-screening pigments in the leaf, including flavonoids, were found in the adaxial epidermal layer, making this layer less transparent to UV-B radiation than the abaxial epidermis, which contained less than 12% of the UV-screening pigments. UV-screening pigments increased by 20% in UV-treated leaves relative to control leaves. Densely arranged epicuticular wax on the adaxial leaf surface of UV-treated plants may have further decreased penetration of UV-B radiation by reflectance. An increased leaf thickness, and decreases in leaf area and leaf dry weight were also found for UV-treated plants.     

Effect of ultraviolet radiation on accumulation and leakage of 86Rb+ in guard cells of Vicia faba

The effects of UV-C (254 nm), UV-A (365 nm) and broad-band UV (280–380 nm) on guard cells of Vicia faba L. cv. Long Pod were investigated in the presence of white light (450 μmol m⁻² s⁻¹). UV-C (7 μmol m⁻² s⁻¹) was found to cause leakage of ⁸⁶Rb⁺ from guard cells, while UV-A (0.3 μmol m⁻² s⁻¹) stimulated increased uptake in these cells. A relatively small stimulatory effect was observed by broad-band UV (3 μmol m⁻² s⁻¹) during the first 30 min of irradiation with an apparent equilibration of influx and efflux thereafter. Leakage of ⁸⁶Rb⁺ from guard cells continued despite the removal of UV-C and an increase in the amount of white light from 450 to 1500 μmol m⁻² s⁻¹, suggesting that membranes were irreversibly damaged. Irradiation of guard cells with UV-C for 30, 45 and 90 min indicated that these cells began to be affected already by 30 min UV-C irradiation.     

Diurnal changes in phosphoenolpyruvate carboxylase and pyruvate, orthophosphate dikinase properties in the natural environment: interplay of light and temperature in a C4 thermophile

Activities of phosphoenolpyruvate (PEP) carboxylase (EC 4.1.1.31) were measured in leaf extracts of field grown Amaranthus paniculatus L. (C₄) during a natural diurnal irradiance and temperature pattern. Enzyme assays were run at both fixed (30°C) and the corresponding leaf temperature at the time of harvest. Light activation of PEP carboxylase (PEPCase) at fixed assay temperatures was expressed as a decrease in S_0–5 (PEP) after a threshold (> 330 μmol m^–2 s^–1) photon fluence rate was surpassed at noon. Earlier in the morning, increase in apparent enzyme affinity for PEP was observed when the assay was run at leaf temperature, indicating a physiologically meaningfull effect of temperature on S^0.5 (PEP). The 3.3-fold increase in PEPCase activity at low PEP and fixed assay temperature between the minimal and maximal irradiance and temperature hours of the day, became 12.8-, 11.5- and 7.4-fold when assays were run at the corresponding leaf temperature during three diurnal cycles with respective temperature differences (max minus min) of 9.0, 8.3 and 7.4°C. The extent of malate inhibition was the same for both day and night forms of PEPCase assayed at 35°C, but increased considerably with night enzyme at 25°C. The results indicate that light increases the apparent affinity of PEPCase for PEP and that at lower temperatures malate becomes more inhibitory. Pyruvate orthophosphate dikinase activity started to increase immediately after sunrise and the 10-fold increase at fixed temperature became 14.8-, 14.2- and 13.1-fold when assays were run at the above leaf temperatures. This indicates that the light effect predominates with pyruvate, orthophosphate dikinase, while with phosphoenolpyravate carboxylase, light and temperature co-operate to increase the day enzyme activities.     

Effects of temperature, pH, and UV radiation on alkaline phosphatase activity in the terrestrial cyanobacterium Nostoc flagelliforme

Cyanobacteria produce phosphatases in response to phosphorus deficiency as some other autotrophs. However, little has been documented on the effects of key climate change factors, such as temperature rise and solar UV radiation (280–400 nm), on cyanobacterial alkaline phosphatase activity. Here, we found that the terrestrial cyanobacterium Nostoc flagelliforme showed higher activity of the enzyme with increasing temperature and pH levels, exhibiting maximal values at 45 °C and pH?11, respectively. However, when exposed to solar radiation in the presence of UV-A (320–400 nm) and UV-B (280–320 nm), significant reduction of the enzyme activity was observed at a photosynthetically active radiation (PAR) level of 300 W?m^?2 (1,450 μmol photons m^?2 s^?1), which is equivalent or lower than the noontime level of solar PAR at the organism's habitats. UV-A and UV-A + UV-B induced about 21 and 39 % inhibition of the enzyme activity in the 3-h exposures. The decrease in the activity of phosphatase can be attributed to the UV radiation-induced inactivation of the enzyme and indirectly to the UV radiation-induced production of reactive oxygen species.     

The responses of a lotic mayfly Nousia sp. (Ephemeroptera: Leptophlebiidae) to moving water and light of different wavelengths

1. Although laboratory studies of the behaviour of aquatic macroinvertebrates are common, there has been little critical evaluation of the importance of test conditions to them. We used a common Australian leptophlebiid mayfly, Nousia sp., to investigate responses to light, wavelength of light, presence or absence of cover and still or flowing water.
2. Nousia sp. showed substantial qualitative differences in behaviour, as measured by movement, when there was no refuge (in the form of a crevice beneath a tile) present in the experimental arena.
3. We found no evidence of diel periodicity in activity in Nousia sp.
4. Nousia sp. did not respond to infra-red, red or green light at a flux density of 18–19 μmol m^–2 s^–1.
5. Nymphs were three times more likely to remain stationary in flowing water (mean velocity 0.10 m s^–1) than in still water.
6. We conclude that generalized assumptions about test conditions for experiments designed to quantify laboratory behaviour in benthic macroinvertebrates are unjustified and that evaluation of the individual requirements of test species should be conducted routinely.     

Photobiological properties of the inhibition of etiolated Arabidopsis seedling growth by ultraviolet-B irradiation

Alteration of 'normal' levels of ultraviolet-B light (UV-B, 280–320 nm) can affect plant chemical composition as well as growth; however, little is known about how plants perceive UV-B light. We have carried out fluence response curves, and demonstrated that the growth inhibition of etiolated Arabidopsis thaliana seedlings by low fluence UV light is specific to UV-B and not UV-A (320–390 nm). The response shows reciprocity between duration and intensity, at least over a limited range, and thus depends only on photon fluence and not on photon flux. The action spectrum for this response indicates a peak of maximum effectiveness at 290 nm, and response spectra at different fluences indicate that the most effective wavelength at 30 000  µ mol m^–2 is 290 nm, whereas 300 nm light was the most effective at 100 000  µ mol m^–2. This response occurs in mutant seedlings deficient in cryptochrome, phytochrome or phototropin, suggesting that none of the known photoreceptors is the major UV-B photoreceptor. Some null mutants in DNA repair enzymes show hypersensitivity to UV-B, suggesting that even at low fluence rates, direct damage to DNA may be one component of the response to UV-B.     

Influence of UV-B radiation on developmental changes, ethylene, CO2 flux and polyamines in cv. Doyenne d'Hiver pear shoots grown in vitro

In vitro shoots of cv. Doyenne ďHiver pear ( Pyrus communis L.) were irradiated under controlled environments for 6 h per day at 5 different levels of biologically effective UV-B radiation (UV-B_BE). UV-B exposure caused a progressive increase in apical necrosis above background levels and stimulated leaf abscission. Shoots grown for 2 weeks at 7. 8 mol m⁻² day ⁻¹ of photosynthetic photon flux (PPF) and treated with 8. 4 or 12. 0 kJ m⁻² day ⁻¹ UV-B_BE produced up to 4 times more ethylene than those given 2. 2 or 5. 1 kJ m⁻² day⁻¹ UV-B_BE or untreated controls. Exposure of shoots to 12 kJ m⁻² day ⁻¹ of UV-B_BE caused an increase in free putreseine content after 4 to 14 days of irradiation. Shoots showed a decrease in CO₂ uptake after 3 days of UV-B: thereafter, they appeared to recover their photosynthetic capacity. Under typical PPF conditions used in micropropagation (90 μmol m⁻² S⁻¹). 8. 4 kJ m⁻² day ⁻¹ of UV-B radiation was injurious to realatively tender tissues of in vitro pear shoots: increasing the level of UV-B_BE to 12 kJ m⁻² day⁻¹ produced even more adverse effects.     

Balancing carboxylation and regeneration of ribulose-1,5- bisphosphate in leaf photosynthesis: temperature acclimation of an evergreen tree, Quercus myrsinaefolia

Changes in the temperature dependence of the photosynthetic rate depending on growth temperature were investigated for a temperate evergreen tree, Quercus myrsinaefolia . Plants were grown at 250 μ mol quanta m^–2 s^–1 under two temperature conditions, 15 and 30 °C. The optimal temperature that maximizes the light-saturated rate of photosynthesis at 350 μ L L^–1 CO₂ was found to be 20–25 and 30–35 °C for leaves grown at 15 and 30 °C, respectively. We focused on two processes, carboxylation and regeneration of ribulose-1,5-bisphosphate (RuBP), which potentially limit photosynthetic rates. Because the former process is known to limit photosynthesis at lower CO₂ concentrations while the latter limits it at higher CO₂ concentrations, we determined the temperature dependence of the photosynthetic rate at 200 and 1000 μ L L^–1 CO₂ under saturated light. It was revealed that the temperature dependence of both processes varied depending on the growth temperature. Using a biochemical model, we estimated the capacity of the two processes at various temperatures under ambient CO₂ concentration. It was suggested that, in leaves grown at low temperature (15 °C), the photosynthetic rate was limited solely by RuBP carboxylation under any temperature. On the other hand, it was suggested that, in leaves grown at high temperature (30 °C), the photosynthetic rate was limited by RuBP regeneration below 22 °C, but limited by RuBP carboxylation above 22 °C. We concluded that: (1) the changes in the temperature dependence of carboxylation and regeneration of RuBP and (2) the changes in the balance of these two processes altered the temperature dependence of the photosynthetic rate.     

The stomatal response to CO2 is linked to changes in guard cell zeaxanthin*

The mechanisms mediating CO₂ sensing and light–CO₂ interactions in guard cells are unknown. In growth chamber-grown Vicia faba leaves kept under constant light (500 μ mol m^–2 s^–1) and temperature, guard cell zeaxanthin content tracked ambient [CO₂] and stomatal apertures. Increases in [CO₂] from 400 to 1200 cm³ m^–3 decreased zeaxanthin content from 180 to 80 mmol mol^–1 Chl and decreased stomatal apertures by 7·0 μ m. Changes in zeaxanthin and aperture were reversed when [CO₂] was lowered. Guard cell zeaxanthin content was linearly correlated with stomatal apertures. In the dark, the CO₂-induced changes in stomatal aperture were much smaller, and guard cell zeaxanthin content did not change with chamber [CO₂]. Guard cell zeaxanthin also tracked [CO₂] and stomatal aperture in illuminated stomata from epidermal peels. Dithiothreitol (DTT), an inhibitor of zeaxanthin formation, eliminated CO₂-induced zeaxanthin changes in guard cells from illuminated epidermal peels and reduced the stomatal CO₂ response to the level observed in the dark. These data suggest that CO₂-dependent changes in the zeaxanthin content of guard cells could modulate CO₂-dependent changes of stomatal apertures in the light while a zeaxanthin-independent CO₂ sensing mechanism would modulate the CO₂ response in the dark.     

Comparison of epilithic and epixylic biofilm development in a boreal river

SUMMARY. 1. We assessed substratum effects on lotic biofilm development by placing glass and white pine sampling units in a fourth-order boreal river, and analysing, at 6-week intervals, upper-surface biofilms for ATP, chlorophyll, ergosterol, and the activities of nine exoenzymes.
2. All parameters, except chlorophyll standing stock (range 80–320 μg dm⁻²) and β-xylosidase activity (range 0.4–4.8 μmol h⁻¹ dm⁻²), were significantly greater for epixylic biofilms than for epilithic ones, but the magnitude of the increases varied from 2 to 5 fold, showing that, even under similar hydrodynamic conditions, epilithic and epixylic biofilms are structurally and functionally distinct. For example, ergosterol concentrations ranged from undetectable to 0.93 μg dm⁻² for epilithon and from 11–49 μg dm⁻² for epixylon; corresponding ranges for ATP were 1.6–3.7 (epilithon) and 4.2–7.7 μg dm⁻² (epixylon), for acid phosphatase activity: 2.3–4.9 and 20–41 μmolh⁻¹dm⁻², and for alkaline phosphatase activity: 1.9–8.1 and 29–150 μmol h⁻¹dm⁻², respectively.
3. The more extensive epixylic development was attributed to utilization of the wood substratum as a supplemental carbon source and to a higher density of microbial attachment sites.     

Light modulation and in vitro effects of adenine nucleotides on leaf nitrate reductase activity in cucumber (Cucumis sativus)

Nitrate reductase (NR, EC 1.6.6.1) activity in attached cucumber ( Cucumis sativus L. cv. Ashley) leaves changed rapidly and reversibly during light/dark transitions, especially when assayed in the presence of free Mg²⁺. Light decreased and darkness increased the sensitivity of the enzyme to inhibition by Mg²⁺. The NR activation state, i.e. activity in the presence of Mg²⁺ relative to activity in the absence of Mg²⁺, increased with light intensity up to 400 μmol m⁻² s⁻¹ PAR (photosynthetically active radiation). When a desalted crude extract from illuminated leaves was preincubated with ATP, NR was gradually inactivated. Inactivation was only observed when activity was assayed in the presence of Mg²⁺. The ATP-inactivated NR remained inactive after removing the excess of ATP by gel filtration and it did not occur in partially purified NR preparations. NR extracted from darkened attached leaves was markedly activated when preincubated with 5'-AMP. These results support the view that inactivation/activation of cucumber-leaf NR in response to light/dark signals most likely involves phosphorylation/dephosphorylation of the enzyme catalysed by endogenous proteins. A substantial activation of NR by preincubation with 5'-AMP was also observed when activity was assayed in the absence of Mg²⁺, thus indicating that 5'-AMP can directly activate NR. Irradiation of an extract from darkened leaves containing FAD promoted a partial activation of NR. This effect was observed both in the +Mg²⁺ and in the −Mg²⁺ assay, indicating that activation was caused by photoexcited flavin and did not involve dephosphorylation of the enzyme.     

Carbon metabolism in germinating Ricinus communis cotyledons. Purification, characterization and developmental profile of NADP-dependent malic enzyme

The possible implication of NADP-dependent malic enzyme (NADP-ME; L-malate:NADP oxidoreductase [oxaloacetate-decarboxylating], EC 1.1.1.40) in fatty acid synthesis was examined in Ricinus communis L. cotyledons, NADP-ME catalyses the conversion of L-malate to pyruvate and NADPH, potential substrates for fatty acid synthesis. NADP-ME activity and protein levels were monitored during germination, up to 20 days postimbibition. The developmental profile showed a peak in activity (6 times with respect to the basal value) and immunoreactive protein (a single 72-kDa band using anti-maize NADP-ME antibodies) around day 7. The enzyme was partially purified (41-fold) and its kinetics characterized. The optimum pH was around 7.1. K_m values for L-malate and NADP⁺ were 0.68 m M and 8.2 μ M respectively. The enzyme used Mg²⁺ or Mn²⁺ as essential cofactors. Several metabolites were assayed as potential enzyme modulators. Succinate, CoA, acetyl-CoA and palmitoyl-CoA were activators of NADP-ME, at saturating or sub-saturating substrate concentrations, K² values for CoA and derivative compounds were in the micromolar range (i.e., 0.8 μ M for acetyl-CoA). No significant effects were obtained with other Krebs cycle intermediates and amino acids (i.e. 2-oxoglutarate, glutamate, glutamine, fumarate). The activity was 29 times higher in the forward (decarboxylating) direction compared to the reverse direction. These results hint at cotyledon NADP-ME behaving as a regulatory enzyme in R. communis . Its activity is responsive to metabolites of the fatty acid synthesis pathway, and thus a role in this metabolism is suggested.     

Photosystem II damage induced by chemically generated singlet oxygen in tobacco leaves

In the present work, we investigated the role of chemically generated singlet oxygen, produced by photodynamic effect of rose bengal, in damaging the PSII complex in tobacco leaves in which protein synthesis-dependent repair was inhibited by infiltration with lincomycin. A 30-min exposure to low-intensity (150 μmol m⁻² s⁻¹) photosynthetically active radiation (PAR) induced singlet oxygen production as detected by quenching of 3-[ N -(β-diethylaminoethyl)- N -dansyl]aminomethyl-2,2,5,5-tetramethyl-2,5-dihydro-1 H -pyrrole fluorescence in leaves infiltrated with both lincomycin and rose bengal. This light treatment caused photoinhibition of PSII, as revealed by the marked loss both of the photochemical yield and the amount of D1 protein in PSII reaction center. When rose bengal was not present in the leaves, these symptoms of photodamage were not induced by the same low-intensity PAR. However, when excitation pressure on PSII was increased to 1500 μmol m⁻² s⁻¹, irreversible photodamage of PSII was also observed, showing that the lincomycin treatment applied in vivo was sufficiently inhibiting protein repair. Our results show that singlet oxygen is able to cause oxidative damage in PSII directly, as suggested earlier and argue against its recently hypothesized role exclusive to inhibiting PSII protein repair ( Nishiyama et al. 2006 ).