Dark-adapted Leaf Research Articles

Role of foliar iron in modulating primary and antioxidant mechanisms of Mentha piperita L. under different light conditions

The present study aims to uncover the role of foliar iron in modulating primary and antioxidant mechanisms of Mentha piperita L. under light conditions. Plants were distributed in a completely randomized design and submitted to five Fe concentrations; 0 (control), 0.5, 1.0, 1.5 and 2.0 g L−1. And under two irradiance conditions; I100% (full sun) and I50% (50% irradiance). Forty-eight hours after foliar Fe application, the following parameters were determined: gas exchange, chlorophyll (a and b), carotenoids and anthocyanin content, chlorophyll a fluorescence, peroxidase and superoxide dismutase activities. The highest Fe concentrations affect the internal CO2 concentration (Ci) and water use efficiency, according to the photochemical extinction coefficient and peroxidase and superoxide dismutase activities. I100% increased the minimum dark-adapted leaf fluorescence and electron transport rate, while, I50% increased the chlorophyll values, and carotenoid content, Ci and transpiration, maximum dark-adapted leaf fluorescence, antenna quantum efficiency and dark-adapted fluorescence. We discovered that isolated treatments are more effective in modifying the response mechanisms of M. piperita plants and the interaction of Fe and irradiance tends to alter antioxidant metabolism.

Measuring Instant Light-Response Curve of Chlorophyll Fluorescence in Sago Palm (Metroxylon sagu Rottb.) Leaves: Different Time Measurements on Dark and Light-Adapted Leaf

This study aimed to determine the appropriate time for the chlorophyll fluorescence light curve measurement in sago palms and to avoid high values variations. The photosynthetic status of three-year-old sago palm seedlings was evaluated through the chlorophyll fluorescence measurement. The chlorophyll fluorescence light curve of measurement was evaluated under different daytime measurements. Observations on DFm/Fm’ and ETR vs. irradiances were conducted on sago palm leaves in the morning, midday, and afternoon with light and dark adaptation. The highest ETRmax value was found under a light-adapted leaf; however, the Eopt value could not be obtained. Morning time measurement on the dark-adapted leaf is the most appropriate method and time to get an Eopt value with high ETRmax.

Interactions between light and temperature during the in vitro as well as in situ modulation of phosphoenolpyruvate carboxylase in leaves of a C3 plant Pisum sativum L.

The C4 phosphoenolpyruvate carboxylase (PEPC) is well known for its modulation by light and temperature. We have attempted to address such coordinated effects of light and temperature on C3 PEPC in leaves of Pisum sativum. Experiments were done with dark-adapted leaf discs as well as with leaves collected at different times during the day and in each month during the year. The modulation of C3 PEPC by temperature was much stronger than that by light. The optimum temperature for in vitro activity of C3 PEPC was 25°C while 20°C was sub-optimal and 30°C to 45°C were supra-optimal. The photo-activation of PEPC was more at 25°C than at 45°C. The activation of C3 PEPC by temperature was more in the dark than in the light. In in situ conditions, a diurnal rhythm in the activity and regulatory properties of C3 PEPC was more prominent at colder than at warm temperatures. However, no consistent trends in the seasonal changes in PEPC activity/ regulatory properties were observed. Scatter plots suggested that light had a greater influence on PEPC activity while temperature exerted a much greater effect on its regulatory properties. Thus, C3 PEPC was also modulated by a natural variation in light and temperature under in situ conditions. These observations correlated well with the light and temperature requirements of C3 plants.

Using Rapid Chlorophyll Fluorescence Transients to Classify Vitis Genotypes

When a dark-adapted leaf is illuminated with saturating light, a fast polyphasic rise of fluorescence emission (Kautsky effect) is observed. The shape of the curve is dependent on the molecular organization of the photochemical apparatus, which in turn is a function of the interaction between genotype and environment. In this paper, we evaluate the potential of rapid fluorescence transients, aided by machine learning techniques, to classify plant genotypes. We present results of the application of several machine learning algorithms (k-nearest neighbors, decision trees, artificial neural networks, genetic programming) to rapid induction curves recorded in different species and cultivars of vine grown in the same environmental conditions. The phylogenetic relations between the selected Vitis species and Vitis vinifera cultivars were established with molecular markers. Both neural networks (71.8%) and genetic programming (75.3%) presented much higher global classification success rates than k-nearest neighbors (58.5%) or decision trees (51.6%), genetic programming performing slightly better than neural networks. However, compared with a random classifier (success rate = 14%), even the less successful algorithms were good at the task of classifying. The use of rapid fluorescence transients, handled by genetic programming, for rapid preliminary classification of Vitis genotypes is foreseen as feasible.

Zinc and potassium fertilizer recommendation for cotton seedlings under salinity stress based on gas exchange and chlorophyll fluorescence responses

Cotton is typically grown in warm regions in which salinity and nutrient deficiency stresses are ubiquitous and often simultaneously influence plant growth. Under saline conditions, fertilizer recommendation is highly challenging, since nutrient application may increase or decrease plant salt tolerance, which may complicate prediction of crop yield. So far, no investigations have been conducted in salt-affected soils to determine optimum concentrations of potassium (K) and zinc (Zn) fertilizers based on chlorophyll fluorescence (ChlF) and gas exchange (GEx) responses in upland cotton (Gossypium hirsutum L.). Accordingly, in this study, a factorial experiment was conducted in a complete block design with six replicates under controlled conditions. Treatments included various K2SO4 (0, 50, 100, and 150 kg ha−1), and ZnSO4 (0, 50, and 100 kg ha−1) concentrations applied to soil before planting. Cottonseeds were sown in non-saline soils and soils formerly salinized with natural saline water diluted to 15 dS m−1. One month after sowing, results showed that salinity significantly decreased dry weight, chlorophyll content index, photosynthesis rate (A), leaf to air vapor pressure, transpiration rate (E), stomatal conductance, and minimum fluorescence of dark-adapted leaf, but increased root to shoot ratio (R/Sh). Under salinity, combined application of K and Zn boosted physiological properties including yield of photosystem II photochemistry (ФPSII), A, and E without improving biomass. Combined application of K and Zn at highest concentrations decreased R/Sh by 93% compared to the control. Rate of increase was higher in E than that of A leading to reduction in water use efficiency. High Zn concentration in saline soils increased non-photochemical quenching and energy loss in form of heat. Under non-saline condition, Zn application significantly decreased A probably due to inhibitory effect on electron transfer within photosystem II. K significantly increased stomatal conductance and accordingly E. GEx parameters were more sensitive to used treatments than ChlF parameters. Based on GEx and ChlF responses, the most salt-tolerant cotton seedlings were obtained under combined application of 50 kg K2SO4 ha−1 and 50 kg ZnSO4 ha−1, thus these concentrations are recommended for optimal establishment of cotton seedlings under salinity stress.

Callus induction and plant regeneration from leaves of peony

Tree peony (Paeonia suffruticosa Andr.) is a valued ornamental plant. This study reports on peony callus induction, shoot organogenesis and plant regeneration using young peony leaves as explants. Various media containing diverse plant growth regulators were assessed for their potency in peony propagation. After exposure of dark-adapted leaf discs to 30 μmol m−2 s−1 of light, inoculation in Murashige and Skoog (MS) + 0.2 mg L−1 2,4-dichlorophenoxyacetic acid (2,4-D) + 0.2 mg L−1 a-naphthaleneacetic acid (NAA) + 3.0 mg L−1 thidiazuron (TDZ) medium resulted to the highest callus induction rate with values reaching up to 87.8%. We identified that MS + 0.2 mg L−1 NAA + 2.0 mg L−1 6-benzyladenine (6-BA) + 2.0 mg L−1 kinetin (KT), with a multiplication coefficient of 3.025, to be the optimal medium for further callus proliferation under light. Inoculation in MS + 2.0 mg L−16-BA + 0.2 mg L−1 NAA + 0.3 mg L−1 TDZ medium allowed 22.22% of callus cultures to differentiate into adventitious shoots, whereas a similar rate of root formation was detected when 1/2 MS + 0.1 mg L−1 NAA + 0.05 mg L−1 IBA + 30 g L−1 sucrose medium was used. Our findings provide important information on peony regeneration and present a new method for peony tissue culture that will potentially facilitate mass propagation and genetic engineering of peony plants.

Performance of wetland forbs transplanted into marshes amended with oil sands processed water.

Companies mining oil sands in Alberta (Canada) face the challenge of reclaiming wetlands under water use restrictions. Wetland reclamation after mining will generate marshes characterized by elevated salinity and residual hydrocarbons. Oil sands wetlands are also impoverished in forbs, suggesting that their establishment may be constrained by water chemistry. We transplanted skullcap, mint, and smartweed plants into experimental trenches that simulated two possible reclamation scenarios: wetlands amended with on-site freshwater or with oil sands processed water (OSPW). The main scientific question was is OSPW a suitable water amendment as freshwater for reclaiming wetland forb habitat? As a surrogate of plant health, we studied plant ecophysiology (gas exchange, leaf fluorescence), leaf chemistry, and plant growth. Results showed that there were no differences in skullcap mineral contents under either treatment; however, mint and smartweed plants subjected to OSPW had a significantly higher Na content than those under freshwater. Smartweed dark-adapted leaf fluorescence showed a reduced photochemistry in OSPW relative to plants in freshwater. Mint leaves exhibited lower stomatal conductance in OSPW than in freshwater, a condition that negatively affected transpiration and carboxylation. Skullcap plants grown in OSPW had lower net CO2 assimilation rates than those in freshwater but did not show any other ecophysiological difference between treatments. Mint plants experienced growth reductions (i.e., shoot height) in OSPW. Our results show, for the first time in the literature, that plants photosynthetic capacity was negatively affected by OSPW. Conditions in OSPW proved to be suitable for establishment as transplanted forbs showed 100% survival after the first growing season. However, impaired physiological functions in plants subjected to OSPW indicated that OSPW amendment created a less hospitable habitat for wetland forbs than freshwater.

Influence of foliar application of silicon on chlorophyll fluorescence, photosynthetic pigments, and growth in water-stressed wheat cultivars differing in drought tolerance

The influence of foliar application of silicon (Si) on chlorophyll contents, chlorophyll fluorescence, and growth of four wheat cultivars differing in drought tolerance (Sirvan and Chamran, as relatively drought tolerant, and Shiraz and Marvdasht, as drought sensitive) was examined under water deficit (100% and 40% F.C.) created in a greenhouse. The results showed that water deficit decreased shoot and root lengths, shoot dry weight, root dry weight, water utilization efficiency, chlorophyll a and b, and chlorophyll stability index. In contrast, foliar application of Si improved plant growth parameters and chlorophyll pigment concentration under water deficit; however, it did not significantly affect wheat growth under control conditions. Limited water supply reduced the values of minimal fluorescence from dark-adapted leaf (F0), maximal fluorescence from dark-adapted leaf (Fm), maximum quantum yield of PSII (Fv/Fm), effective quantum yield of PSII (PSII), photochemical quenching (qP), and apparent photosynthetic electron transport rate (ETR). However, under water deficit, foliar application of Si application increased the earlier mentioned parameters. In contrast, nonphotochemical quenching (qN) and F0/ Fm increased under water deficit, and application of Si further improved these parameters. Chlorophyll fluorescence analysis suggested that Si alleviated water deficit-induced adverse effects by reducing nonphotochemical quenching, while increasing Fv/Fm and qP, so that it improved the light use efficiency in the four wheat cultivars under stress. Overall, we concluded that drought-sensitive cultivars (Shiraz and Marvdasht) could resemble resistant cultivars upon foliar application of silicon.

Thermal phase and excitonic connectivity in fluorescence induction

Chl fluorescence induction (FI) was recorded in sunflower leaves pre-adapted to darkness or low preferentially PSI light, or inhibited by DCMU. For analysis the FI curves were plotted against the cumulative number of excitations quenched by PSII, n q, calculated as the cumulative complementary area above the FI curve. In the +DCMU leaves n q was <1 per PSII, suggesting pre-reduction of Q A during the dark pre-exposure. A strongly sigmoidal FI curve was constructed by complementing (shifting) the recorded FI curves to n q=1 excitation per PSII. The full FI curve in +DCMU leaves was well fitted by a model assuming PSII antennae are excitonically connected in domains of four PSII. This result, obtained by gradually reducing Q A in PSII with pre-blocked Q B (by DCMU or PQH2), differs from that obtained by gradually blocking the Q B site (by increasing DCMU or PQH2 level) in leaves during (quasi)steady-state e(-) transport (Oja and Laisk, Photosynth Res 114, 15-28, 2012). Explanations are discussed. Donor side quenching was characterized by comparison of the total n q in one and the same dark-adapted leaf, which apparently increased with increasing PFD during FI. An explanation for the donor side quenching is proposed, based on electron transfer from excited P680* to oxidized tyrosine Z (TyrZ(ox)). At high PFDs the donor side quenching at the J inflection of FI is due mainly to photochemical quenching by TyrZ(ox). This quenching remains active for subsequent photons while TyrZ remains oxidized, following charge transfer to Q A. During further induction this quenching disappears as soon as PQ and Q A become reduced, charge separation becomes impossible and TyrZ is reduced by the water oxidizing complex.

Effect of sulphate nutrition on arsenic translocation and photosynthesis of rice seedlings

The effect of sulphate nutrition on arsenic (As) concentration, photosynthetic and chlorophyll fluorescence parameters of rice was investigated in hydroponically grown rice seedlings (Oryza sativa L.), using three sulphate levels (1.8 μM, 0.7 mM, or 1.5 mM). The results showed that sulphate deficiency decreased As accumulation in root, but increased the translocation of As from root to shoot. Sulphate deficiency reduced maximum quantum yield (Fv/Fm), minimum fluorescence and electron transport rate (ETR) of a dark-adapted leaf. Compared with low sulphate treatments (1.8 μM), significant increases were observed in the parameters of rapid light curves, rETRmax and Ik of photosystem I (PSI) and photosystem II (PSII) of rice grown in the high sulphate treatments (1.5 mM) regardless of As additions. Therefore, an adequately high sulphate supply may result in less As translocation from root to shoot, and protecting the reaction pathways of PSI and PSII of rice seedlings grown in higher As-contaminated medium.

Dark-adapted leaf conductance, but not minimum leaf conductance, predicts water use efficiency of soybean (Glycine max L. Merr.)

Walden-Coleman, A. E., Rajcan, I. and Earl, H. J. 2013. Dark-adapted leaf conductance, but not minimum leaf conductance, predicts water use efficiency of soybean (Glycine max L. Merr.). Can. J. Plant Sci. 93: 13–22. The conductance to water vapor of dark-adapted leaves (gdark) has been shown to be negatively correlated with whole-plant water use efficiency (WUE) in soybean, but the physiological basis of this relationship is unknown. It is also not clear how gdark compares with the minimum leaf conductance of wilted leaves (gmin), a trait that has been studied extensively across a broad range of species. We compared gdark to gmin of soybean leaves and found that gdark values were consistently much higher than gmin values measured on the same leaves. Also, across seven soybean varieties known to differ for WUE, gdark but not gmin was correlated with WUE. Thus, gdark and gmin should be considered distinct traits. We measured gdark at two different leaf positions, and found that gdark measured at the lower leaf position (two main stem nodes below the youngest fully expanded leaf) was best correlated with WUE. We then used this method to screen a selection of current commercial soybean varieties adapted to Ontario, Canada, for variation in gdark. The range in gdark among the commercial varieties was as broad as that measured previously among more diverse genotypes, suggesting that Ontario soybean varieties might also vary widely for WUE.

Dark-adapted leaf conductance, but not minimum leaf conductance, predicts water use efficiency of soybean (Glycine max L. Merr.)

Walden-Coleman, A. E., Rajcan, I. and Earl, H. J. 2013. Dark-adapted leaf conductance, but not minimum leaf conductance, predicts water use efficiency of soybean (Glycine max L. Merr.). Can. J. Plant Sci. 93: 13-22. The conductance to water vapor of dark-adapted leaves (gdark) has been shown to be negatively correlated with whole-plant water use efficiency (WUE) in soybean, but the physiological basis of this relationship is unknown. It is also not clear how gdark compares with the minimum leaf conductance of wilted leaves (gmin), a trait that has been studied extensively across a broad range of species. We compared gdark to gmin of soybean leaves and found that gdark values were consistently much higher than gmin values measured on the same leaves. Also, across seven soybean varieties known to differ for WUE, gdark but not gmin was correlated with WUE. Thus, gdark and gmin should be considered distinct traits. We measured gdark at two different leaf positions, and found that gdark measured at the lower l...

Early detection of drought stress in tomato plants with chlorophyll fluorescence imaging–practical application of the speaking plant approach in a greenhouse–

AbstractThe chlorophyll fluorescence imaging technique is useful for evaluating photosynthetic functions of plants without actually touching the plant. In our previous study, we developed a chlorophyll fluorescence imaging system for tomato plants cultivated in greenhouses. This imaging system measures the chlorophyll fluorescence induction phenomenon, a dynamic change in chlorophyll fluorescence intensity induced by illuminating a dark-adapted leaf with a stable intensity excitation light, and analyzes the shape of the induction curve, i.e., the temporal course of chlorophyll fluorescence intensity during this phenomenon. The shape of the induction curve is characterized by an initial maximum peak (P), subsequent transient dip (S), and secondary small peak (M). We defined an index, the photosynthetic function index (PFI; fluorescence intensity of P divided by the average fluorescence intensity from S to M), to evaluate the shape of the induction curve. In this study, we applied this system to detect drought stress in tomato plants cultivated in a semi-commercial greenhouse. PFI was clearly lower in stressed plants than in healthy plants. The decreased PFI in stressed plants is probably attributable to photosynthetic dysfunction in these plants.

Observation of plastoquinone kinetics in photosystem II from delayed fluorescence measurements

Attempts to account for the variations in photosystem II (PSII) under general conditions result in non-linear and cumbersome models that are difficult to validate and render few insights about the system kinetics. In this research, the authors experimentally show that under certain conditions, linear-system techniques could be applied to advantage for probing some basic kinetic characteristics of the plastoquinones (PQs). The PQ redox states of the reaction centres were represented in a conditionally linear model structure with delayed fluorescence (DF) as a measurable output. DF data were acquired for different plant samples and conditions. After least-squares parameter optimisation, not only could the model closely describe the measured DF, but more significantly, the estimated parameters correctly reflected the expected changes induced by drought or [3-(3,4-dichlorophenyl)-1,1-dimethylurea] (DCMU) stress. Analysis showed that for short-pulse illumination, the PQ kinetic states of the reaction centres in an initially dark-adapted plant leaf can be represented as a time-invariant bilinear system in a five-dimensional state space. The system becomes linear for constant illuminations, but the system matrix and the kinetic behaviour are illumination dependent. In particular, the system behaves differently between lights-on and lights-off conditions. The simplicity of the model structure, nonetheless, permits observation and analysis of the PQ kinetics of PSII reaction centres from DF measurements by using linear-system techniques.

Comparison of thermostability of PSII between the chromatic and green leaf cultivars of Amaranthus tricolor L.

In the present study, we investigated the antioxidative potential in leaves of the chromatic (CC) versus green (GC) Amaranthus tricolor L. under moderate high-temperature stress at 45°C. Before heat stress, CC had significantly higher levels of betacyanins [about 3.2 mg g−1(FM)] than the green [1.8 mg g−1(FM) (p<0.01), while similar chlorophyll (Chl) content [about 2 mg g−1(FM)] was observed between both cultivars. After exposure to high temperature (45°C) for 6 days, betacyanins in leaves of CC were remarkably increased (about 2 times of that in control samples grown at 30°C). In contrast, betacyanins in GC significantly decreased by 56% in comparison with that of the control. Chl level in CC was higher than that in GC after heat stress for 6 days. Flavonoids and total phenolics in both cultivars were increased, but much more in CC. Significantly less H2O2 accumulation was observed in the leaves and stems of CC than in those of GC under heat stress. Interestingly, much stronger circadian oscillation in fluorescence was observed in both cultivars after treatment at 45°C, which suggested that heat stress stimulates endogenous rhythms of photosystem II (PSII). Under moderate high-temperature stress, Chl fluorescence parameters Fv/Fm (maximum quantum yield of PSII), qP (coefficient of photochemical quenching), ΦPSII (effective PSII quantum yield), and ETR (electron transport rate) exhibited a gradual decrease, NPQ (nonphotochemical quenching) showed a slight increase followed by a gradual decline, whereas Fo (minimum fluorescence of a dark-adapted leaf) increased continuously. In contrast to GC, after 120 h of high-temperature treatment, CC exhibited significantly lower Fo level, and higher levels of Fv/Fm and NPQ. It is clear that PSII in CC was more stable than that in GC. The results indicate that betacyanins are an effective antioxidant, and probably contribute greatly to the higher thermal stability of PSII and higher tolerance to heat stress.

Leaf Chlorophyll Fluorescence: Background and Fundamentals for Plant Biologists

This review on chlorophyll a fluorescence starts with an overview of the primary photochemistry occurring at PSII and a characterization of the so-called “open” and “closed” states of its reaction centers. This provides the theoretical background for understanding the origin of PSII-emitted fluorescence and how its yield varies with the fraction of open reaction centers. The review proceeds to discuss the changes in fluorescence emission following illumination of a dark-adapted leaf and to define the PSII intrinsic quantum yield of photochemistry, which in turn provides an indication of PSII capacity. In light-adapted leaves, it is discussed how the use of modulated fluorometers and the double lighting technique allow an evaluation of photochemical and non-photochemical quenching, two parameters that give useful information about the plant’s photosynthetic performance under field conditions. Finally, it is described how the PSII operational efficiency can be used to calculate the photosynthetic electron transport rate and the conditions under which this is linearly related to the CO2 assimilation rate. Some requirements for a valid application of the technique as well as some limitations in interpreting its results are discussed.

A comparative study of oak (Quercus, Fagaceae) seedling physiology during summer drought in southern California

Natural recruitment of oaks appears to be declining throughout the northern hemisphere. Summer drought poses a potentially important barrier to oak recruitment in southern California. To evaluate this barrier, we grew evergreen Quercus agrifolia and deciduous Q. lobata from seeds near parental trees. We measured water relations, chlorophyll fluorescence, and gas exchange during these seedlings' fourth and fifth summers and compared them to neighboring adults. Most seedlings had substantially lower values for predawn xylem pressure potential (Ψ(pd)), minimum photosystem II (PSII) quantum efficiency (Φ(PSIIMIN)), maximum quantum efficiency for PSII under dark-adapted leaf conditions (Fv/Fm), and maximum photosynthetic assimilation (Amax), and higher values for maximum nonphotochemical quenching (NPQmax) than did conspecific adults. The high, unvarying Ψ(pd) values of the adults suggest they use perennially available groundwater. Quercus lobata seedlings commonly had lower values for Ψ(pd) than did Q. agrifolia, and values for Ψ(pd) and Φ(PSIIMIN) were significantly related to size in Q. lobata but not in Q. agrifolia. These data suggest important interspecific differences in root architecture. Lower values for Φ(PSIIMIN), Fv/Fm, and higher NPQmax in Q. agrifolia indicate that Q. agrifolia seedlings were usually under more stress than Q. lobata, which typically had higher Amax rates than did Q. agrifolia seedlings. Diurnal photosynthesis curves were quite flat for Q. agrifolia, but they peaked in the morning for Q. lobata. Established seedlings appeared to be under more stress than adults, but this stress did not appear severe enough to cause death. Access to perennially available groundwater may be crucial for the seedling to sapling transition.

Influence of brick air scrubber by-product on growth and development of corn and hybrid poplar

Studies were conducted to determine the effects of spent reagent from air pollution control scrubbers used at a brick manufacturing facility on emergence, growth, and physiological responses of corn and hybrid poplar plants. Scrubber by-product was obtained from General Shale Brick, Louisville, KY. Potting substrate was weighed and quantities of scrubber by-product were added to the substrate to obtain treatments of 0%, 6.25%, 12.5%, 25%, 50%, 75%, and 100% scrubber by-product (w:w) for the corn study. Each treatment mix was potted into nine replicate polyethylene pots and four corn seeds were sown per pot. The pots were randomized in a greenhouse at Clemson University and the number of seedlings emerging from each treatment, dark-adapted leaf chlorophyll a fluorescence, and shoot heights were measured at the end of a 21-day growth period. Then, dry shoot biomass was determined for plants from each treatment and plant tissues were analyzed for selected constituents. For the poplar study, nine-inch cuttings of hybrid poplar clone 15–29 ( Populus trichocarpa × P. deltoides) and clone OP367 ( P. deltoides × P. nigra) were planted in treatments of scrubber by-product-potting soil mixes of 0% , 5% , 10% , and 25% w:w. Leaf chlorophyll a fluorescence was measured over six weeks and cumulative leaf area, dry biomass, and nutrient content of tissues were determined upon harvest. Results of these studies indicate that percent seedling emergence for corn plants decreased with increasing scrubber by-product application rates. Application rates up to 12.5% scrubber by-product w:w had no adverse effect on corn seedling emergence. Shoot elongation, biomass production, and the status of the photosynthetic apparatus of the seedlings were also not severely impaired at applications below this level. A critical value of 58.2% w:w scrubber by-product was estimated to cause 25% inhibition of seedling emergence. Biomass production, cumulative leaf area, and chlorophyll a fluorescence of hybrid poplar plants were not affected by scrubber by-product applications of up to 5% w:w.

A thermoluminescence study of Photosystem II back electron transfer reactions in rice leaves – effects of salt stress

The recombination reactions of Photosystem II have been investigated in vivo in rice leaves by using the thermoluminescence (TL) emission technique. Excitation of dark-adapted leaf segments at 0 degrees C with different number of single turn-over flashes induced the appearance of complex TL glow curves. The mathematical analysis of these curves showed the existence of four TL components: B1-band (temperature maximum, t(max), at 24 degrees C, originating from S3QB - recombination), B2-band (tmax at 35 degrees C, from S2QB -), AG-band (tmax at 46 degrees C) and C-band (tmax at 55 degrees C, from TyrD +QA -). Their contributions to the total TL signal were different depending on the number of flashes given. AG-band seems to reflect a special electron transfer from some unknown stroma donor to PS II. Q-band (tmax at 19 degrees C), originating from S2QA - recombination, was recorded after flashing samples incubated in the presence of DCMU. The recombination halftimes (t1/2) at 20 degrees C of S2QA -, S3QB -, S2QB - and TyrD +QA - were, respectively, 0.8 s, 48 s, 74 s and about 1 h. A sharp AG-band (tmax at 50 degrees C and t1/2 of 210 s) could be also observed after illumination of leaves with far-red light and after a dark incubation period of whole plants. Incubation of leaf segments with 0.5 M NaCl abolished the inductions of AG-band by darkness and far-red illumination, significantly decreased Q-band intensity, whereas induced a strong increase in C-band intensity. The possible inhibition of S2/S3 formation and quinone oxidation by saline stress are discussed.

Modulation of phosphoenolpyruvate carboxylase in vivo by Ca 2+ in Amaranthus hypochondriacus, a NAD-ME type C 4 plant: Possible involvement of Ca 2+ in up-regulation of PEPC-protein kinase in vivo

The properties of phosphoenolpyruvate carboxylase (PEPC) were studied, with respect to calcium (Ca2+), in leaves of Amaranthus hypochondriacus, a C4 plant. Experiments were conducted in vitro (by adding Ca2+ during enzyme assay) or in vivo (by feeding Ca2+ to intact leaves through petiole). Inclusion of 10 microM Ca2+ during assay marginally increased (<30%) malate sensitivity of PEPC in extracts from dark-adapted leaves. The effect of Ca2+ was marginal on PEPC in extracts from illuminated leaves. Upon applying a low concentration of Ca2+ to leaves, the PEPC activity in leaves increased by 1.5-fold, while inhibition by malate decreased markedly. The light activation of PEPC in Ca2+-fed leaves was slightly higher than in the absence of Ca2+-ethyleneglycol-bis-(beta-aminoethyl ether) N,N,N',N'-tetra acetic acid (EGTA). To assess further the role of Ca2+, 5 mM EGTA (Ca2+ chelator) was either added during the enzyme assay or fed to leaves through petiole. EGTA had no effect on PEPC, when added during enzyme assay. Upon feeding EGTA, the PEPC activity in the dark-adapted leaf extracts increased by 30%, and the effect on malate sensitivity was marginal. However, there was a decrease in PEPC activity in illuminated extracts, resulting in a marked decrease in the extent of light activation of PEPC. The extent of phosphorylation of PEPC was much higher in Ca2+ or Ca2+-EGTA-fed leaves than in the control, but EGTA decreased the light-induced phosphorylation. Our results suggest that optimal alone concentration of Ca2+ is essential for PEPC in leaves of A. hypochondriacus, particularly in vivo. We suggest that Ca2+ regulates PEPC, at an upstream level, such as transcription, by modulating PEPC-protein kinase, thus facilitating the light activation of PEPC.