Temperature Dependence of the Linkage of Quantum Yield of Photosystem II to CO2 Fixation in C4 and C3 Plants

The temperature dependence of quantum yields of electron transport from photosystem II (PSII) ([phi]II, determined from chlorophyll a fluorescence) and CO2 assimilation ([phi]CO2, apparent quantum yield for CO2 assimilation) were determined simultaneously in vivo. With C4 species representing NADP-malic enzyme, NAD-malic enzyme, and phosphoenolpyruvate carboxykinase subgroups, the ratio of [phi]II/[phi]CO2 was constant over the temperature range from 15 to 40[deg]C at high light intensity (1100 [mu]mol quanta m-2 s-1). A similar response was obtained at low light intensity (300 [mu]mol quanta m-2 s-1), except the ratio of [phi]II/[phi]CO2 increased at high temperature. When the true quantum yield for CO2 fixation ([phi]CO2*) was calculated by correcting for respiration in the light (estimated from temperature dependence of dark respiration), the ratio of [phi]II/[phi]C02* remained constant with varying temperature and under both light intensities in all C4 species examined. Because the [phi]II/[phi]CO2* ratio was the same in C4 monocots representing the three subgroups, the ratio was not affected by differences in the bio-chemical mechanism of concentrating CO2 in the bundle sheath cells. The results suggest that PSII activity is closely linked to the true rate of CO2 fixation in C4 plants. The close relationship between [phi]II and [phi]CO2* in C4 species under varying temperature and light intensity conditions is apparently due to a common low level of photorespiration and a primary requirement for reductive power in the C3 pathway. In contrast, in a C3 plant the [phi] II/[phi]CO2* ratio is higher under normal atmospheric conditions than under nonphotorespiratory conditions and it increases with rising temperature. This decrease in efficiency in utilizing energy derived from PSII for CO2 fixation is due to an increase in photorespiration. In both the C3 and C4 species, photochemistry is limited under low temperature, and thus excess energy must be dissipated by nonphotochemical means.     

Estimation of Bundle Sheath Cell Conductance in C4 Species and O2 Insensitivity of Photosynthesis

Low conductance to CO2 of bundle sheath cells is required in C4 photosynthesis to maintain high [CO2] at the site of ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco). Elevated [CO2] allows high CO2 assimilation rates by this enzyme and prevents Rubisco oxygenase activity and O2 inhibition of carboxylation. Bundle sheath conductance to CO2 was estimated by chemically inhibiting phosphoenolpyruvate carboxylase and calculating the slope of the linear response of leaf CO2 uptake to [CO2]. The inhibitor 3,3-dichloro-2-dihydroxyphosphinoylmethyl-2-propenoate was supplied to detached leaves of Panicum maximum, Panicum miliaceum, and Sorghum bicolor at 4 mM. Uptake of CO2 was measured at 210 mL L-1 O2 over the CO2 concentration range of 0.34 to 28 mL L-1. Without the inhibitor, CO2 uptake increased steeply at low [CO2] and saturated at about 1 mL L-1. After inhibition, CO2 uptake was a linear function of [CO2] over much of the range tested. The slope of this CO2 response, taken as bundle sheath conductance, was 2.35, 1.96, and 1.13 mmol m-2 s-1 for P. maximum, P. miliaceum, and S. bicolor, respectively, on a leaf area basis. Conductance based on bundle sheath area was 0.76, 0.93, and 0.54 mmol m-2 s-1, respectively. Uptake of CO2 by leaves of P. maximum supplied with the inhibitor was not affected by reduction of [O2] from 210 to 20 mL L-1 over the range of [CO2] used. Because [CO2] in bundle sheath cells of inhibited leaves is likely to be much lower than ambient, the lack of O2 sensitivity of CO2 uptake cannot be ascribed to lack of O2 reaction with ribulose bisphosphate and is probably due to the low conductance of bundle sheath cells, especially at low ambient [CO2]. The likely result of reducing [O2] from 210 to 20 mL L-1 is to stimulate carboxylation of ribulose bisphosphate, thus further reducing [CO2] in bundle sheath cells and increasing CO2 diffusion to these cells from the mesophyll. However, the increase in diffusion is greatly limited by low conductance of the bundle sheath cell walls. Calculations based on estimated bundle sheath conductance show that changes in bundle sheath [CO2] of 0.085 to 0.5 mL L-1, which might be associated with reduced [O2], would have a negligible effect on CO2 uptake.     

C4 Photosynthesis (The CO2-Concentrating Mechanism and Photorespiration)

Despite previous reports of no apparent photorespiration in C4 plants based on measurements of gas exchange under 2 versus 21% O2 at varying [CO2], photosynthesis in maize (Zea mays) shows a dual response to varying [O2]. The maximum rate of photosynthesis in maize is dependent on O2 (approximately 10%). This O2 dependence is not related to stomatal conductance, because measurements were made at constant intercellular CO2 concentration (Ci); it may be linked to respiration or pseudocyclic electron flow. At a given Ci, increasing [O2] above 10% inhibits both the rate of photosynthesis, measured under high light, and the maximum quantum yield, measured under limiting light ([phi]CO2). The dual effect of O2 is masked if measurements are made under only 2 versus 21% O2. The inhibition of both photosynthesis and [phi]CO2 by O2 (measured above 10% O2) with decreasing Ci increases in a very similar manner, characteristically of O2 inhibition due to photorespiration. There is a sharp increase in O2 inhibition when the Ci decreases below 50 [mu]bar of CO2. Also, increasing temperature, which favors photorespiration, causes a decrease in [phi]CO2 under limiting CO2 and 40% O2. By comparing the degree of inhibition of photosynthesis in maize with that in the C3 species wheat (Triticum aestivum) at varying Ci, the effectiveness of C4 photosynthesis in concentrating CO2 in the leaf was evaluated. Under high light, 30[deg]C, and atmospheric levels of CO2 (340 [mu]bar), where there is little inhibition of photosynthesis in maize by O2, the estimated level of CO2 around ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) in the bundle sheath compartment was 900 [mu]bar, which is about 3 times higher than the value around Rubisco in mesophyll cells of wheat. A high [CO2] is maintained in the bundle sheath compartment in maize until Ci decreases below approximately 100 [mu]bar. The results from these gas exchange measurements indicate that photorespiration occurs in maize but that the rate is low unless the intercellular [CO2] is severely limited by stress.     

Expressing an RbcS Antisense Gene in Transgenic Flaveria bidentis Leads to an Increased Quantum Requirement for CO2 Fixed in Photosystems I and II

It was previously shown with concurrent measurements of gas exchange and carbon isotope discrimination that the reduction of ribulose-1,5-bisphosphate carboxylase/oxygenase by an antisense gene construct in transgenic Flaveria bidentis (a C4 species) leads to reduced CO2 assimilation rates, increased bundle-sheath CO2 concentration, and leakiness (defined as the ratio of CO2 leakage to the rate of C4 acid decarboxylation; S. von Caemmerer, A. Millegate, G.D. Farquhar, R.T. Furbank [1997] Plant Physiol 113: 469-477). Increased leakiness in the transformants should result in an increased ATP requirement per mole of CO2 fixed and a change in the ATP-to-NADPH demand. To investigate this, we compared measurements of the quantum yield of photosystem I and II ([phi]PSI and [phi]PSII) with the quantum yield of CO2 fixation ([phi]CO2) in control and transgenic F. bidentis plants in various conditions. Both [phi]PSI/[phi]CO2 and [phi]PSII/[phi]CO2 increased with a decrease in ribulose-1,5-bisphosphate carboxylase/oxygenase content, confirming an increase in leakiness. In the wild type the ratio of [phi]PSI to [phi]PSII was constant at different irradiances but increased with irradiance in the transformants, suggesting that cyclic electron transport may be higher in the transformants. To evaluate the relative contribution of cyclic or linear electron transport to extra ATP generation, we developed a model that links leakiness, ATP/NADP requirements, and quantum yields. Despite some uncertainties in the light distribution between photosystem I and II, we conclude from the increase of [phi]PSII/[phi]CO2 in the transformants that cyclic electron transport is not solely responsible for ATP generation without NADPH production.     

Regulation of Electron Transport in Photosystems I and II in C3, C3-C4, and C4 Species of Panicum in Response to Changing Irradiance and O2 Levels

Regulation of the quantum yields of linear electron transport and photosystem II photochemistry ([phi]II) with changing irradiance and gas-phase O2 concentration was studied in leaf tissue from Panicum bisulcatum (C3), Panicum milioides (C3-C4), and Panicum antidotale (C4) at 200 [mu]bars of CO2 and 25[deg]C using infrared gas analysis and chlorophyll fluorescence yield measurements. When the O2 level was increased from 14 to 213 mbars at high irradiance, [phi]II increased by as much as 115% in P. bisulcatum but by no more than 17% in P. antidotale. Under the same conditions [phi]II increased to an intermediate degree in P. milioides. Measurements of accumulation of the photooxidized form of the photosystem I reaction center (P700+) based on the light-dependent in vivo absorbance change at 830 nm indicate that the steady-state concentration of P700+ varied in an antiparallel manner with [phi]II when either the irradiance or O2 concentration was changed. Hence, O2-dependent changes in [phi]II were indicative of variations in linear photosynthetic electron transport. These experiments revealed, however, that a significant capacity was retained for in vivo regulation of the apparent quantum yield of photosystem I ([phi]I) independently of [phi]II+ Coordinate regulation of quantum yields of photosystems I and II (expressed as [phi]I:[phi]II in response to changing irradiance and O2 level differed markedly for the C3 and C4 species, and the response for the C3-C4 species most closely resembled that observed for the C4 species. The fraction of total linear electron transport supporting photorespiration at 213 mbars of O2 was negligible in the C4 species and was 13% lower in the C3-C4 species relative to the C3 species as calculated from fluorescence and gas-exchange determinations. At high photon-flux rates and high O2 concentration, the potential benefit to light use for net CO2 uptake arising from lower photorespiration in P. milioides was offset by a reduced capacity for total CO2- and O2-dependent noncyclic electron transport in this species compared with P. bisulcatum.     

Aluminium modulation of the photosynthetic carbon reduction cycle in Zea mays

Two-weeks-old maize (Zea mays L. cv. XL-72.3) plants were submitted to Al concentrations of 0-81 g m-3 for 20 d, after which the A1 concentration-dependent effects on CO2 uptake by the mesophyll tissue and subsequent CO2 assimilation in the photosynthetic carbon reduction cycle of bundle sheath cells were investigated. The net photosynthetic rate (PN) and stomatal conductance (gs) increased continuously up to 27 g m-3 Al, whereas the intercellular CO2 concentration showed minimum values with the 27 g m-3 Al treatment. Moreover, the starch and saccharide concentrations, and fructose-1,6-bisphosphatase did not change significantly with increasing Al concentrations. The photosynthetic electron transport rates along with photosystems 2 and 1 started falling from 9 g m-3 Al onwards, while thylakoid acyl lipid composition did not show a clear pattern. With the Al concentration at 81 g m-3, NADP-malate dehydrogenase activity decreased to minimum values, whereas the opposite occurred with those of pyruvate dikinase, NADP-malic enzyme, and phosphoenolpyruvate carboxylase. Thus in vivo Al concentrations modulate the photosynthetic reduction cycle, possibly by interacting with the carbon flow rate exported to the cytosol. Although the inhibition of NADP-malate dehydrogenase activity might limit pyruvate dikinase, NADP-malic enzyme, and phosphoenolpyruvate carboxylase activities, in vivo the balance between phosphoenolpyruvate production and its carboxylation remains unaffected.     

Aluminium modulation of the photosynthetic carbon reduction cycle in Zea mays

Two-weeks-old maize (Zea mays L. cv. XL-72.3) plants were submitted to Al concentrations of 0-81 g m-3 for 20 d, after which the A1 concentration-dependent effects on CO2 uptake by the mesophyll tissue and subsequent CO2 assimilation in the photosynthetic carbon reduction cycle of bundle sheath cells were investigated. The net photosynthetic rate (PN) and stomatal conductance (gs) increased continuously up to 27 g m-3 Al, whereas the intercellular CO2 concentration showed minimum values with the 27 g m-3 Al treatment. Moreover, the starch and saccharide concentrations, and fructose-1,6-bisphosphatase did not change significantly with increasing Al concentrations. The photosynthetic electron transport rates along with photosystems 2 and 1 started falling from 9 g m-3 Al onwards, while thylakoid acyl lipid composition did not show a clear pattern. With the Al concentration at 81 g m-3, NADP-malate dehydrogenase activity decreased to minimum values, whereas the opposite occurred with those of pyruvate dikinase, NADP-malic enzyme, and phosphoenolpyruvate carboxylase. Thus in vivo Al concentrations modulate the photosynthetic reduction cycle, possibly by interacting with the carbon flow rate exported to the cytosol. Although the inhibition of NADP-malate dehydrogenase activity might limit pyruvate dikinase, NADP-malic enzyme, and phosphoenolpyruvate carboxylase activities, in vivo the balance between phosphoenolpyruvate production and its carboxylation remains unaffected.     

Salinity and salt composition effects on seed germination and root length of four sugar beet cultivars

Salinization is one of the most important factors affecting agricultural land in the world. Salinization occurs naturally in arid and semiarid regions where evaporation is higher than rainfall. Sugar beet yield declines with an increase in salinity, but the sensitivity to salts varies with salt composition in water and sugar beet growth stage. The aim of this study was to determine the effect of water salinity levels and salt composition on germination and seedling root length of four sugar beet cultivars (PP22, IC2, PP36, and 7233). The experiments were undertaken with irrigation water with two salt compositions (NaCl alone and mixture of MgSO₄ + NaCl + Na₂SO₄ + CaCl₂) in three replicates. Thirteen salinity levels with electrical conductivity (EC) of the irrigation water ranging from 0 to 30 dS/m were applied to each cultivar in both experiments. Seed germination percentage and seedling root length growth were determined in 13 days. Statistical analysis revealed that germination and root length were significantly affected by salt composition, cultivars and salinity levels. Regardless of salt composition, seed germination and seedling root length were significantly affected by the irrigation water with EC up to 8 dS/m and 4 dS/m, respectively. Except for cultivar PP22, the adverse effect of salinity of the irrigation water on seed germination and seedling root length was higher for NaCl alone than for the salt mixture, which refers to lower salt stress in field conditions with natural salt composition. Presented at the International Conference on Bioclimatology and Natural Hazards, Poľana nad Detvou, Slovakia, 17–20 September 2007.     

The response of lucerne (Medicago sativa L.) to sodium sulphate and chloride salinity

Sodium and sulphate-dominated salinity is a serious environmental problem occurring in soils and groundwater in many parts of the world. The effect of Na2SO4 and NaCl, at electrical conductivity levels ranging from 2 to 17 dS m-1, on the growth and tissue ion concentrations of 16 lines of lucerne (Medicago sativa L.) was examined in the greenhouse over a 2 month period. Averaged across all lines, plants grown at 17 dS m-1 produced 66% of the dry matter of plants grown at 2 dS m-1. However there were significant differences among lines in relative salt tolerance (as defined by the slope of the reduction in dry matter) versus electrical conductivity. Dry matter production was negatively correlated with shoot concentrations of Na+, Cl- and S2- and generally lines that were more tolerant to salinity had lower concentrations of those ions in the shoots. We conclude that lucerne is moderately tolerant to Na2SO4 -predominated salinity, and that the degree of intraspecific variation that exists within this species will allow more tolerant lines to be selected for establishment in conditions where sulphate salinity is a problem.Collaborator     

Grafting influence on the weight and quality of tomato fruit under salt stress

Two commercial tomato cultivars were used to determine whether grafting could prevent decrease of fruit weight and quality under salt stress conditions. The cultivars Buran F1 and Berberana F1 were grafted onto rootstock ‘Maxifort’ and grown under three levels of elevated soil salinity (EC 3.80 dS m^?1, 6.95 dS m^?1 and 9.12 dS m^?1). Fruit weight reduction of grafted plants was lower (about 20–30%) in comparison with non‐grafted ones. Salt stress at the second salinity level (EC 6.95 dS m^?1) induced the highest alteration of examined growth and quality parameters. The total increase of phenols, flavonoids, ascorbate and lycopene content in the fruits of both grafted and non‐grafted plants for both cultivars had a similar trend and intensity, though some inter‐cultivar variation was observed. The possibility of grafting tomato plants to improve salt tolerance without fruit quality loss is discussed.     

Leaf development,transpiration and ion uptake and distribution in sugarcane cultivars grown under salinity

The effects of salinity on leaf growth, initiation and senescence, on transpiration rates, on leaf water potential and on uptake and distribution of several ions were studied in two sugarcane cultivars differing in salinity sensitivity. Plants, growing in a growing mixture in pots, were exposed to salinized irrigation water for 68 days, starting 60 days after planting. EC values of the irrigation water were 1.0, 2.0, 4.0, 8.0 and 12 dS/m, obtained by using a mixture of NaCl and CaCl2. Plants were also grown in nutrient solution and were at a similar age when exposed to a salinity level of 3 dS/m for 30 days followed by 6.0 dS/m for an additional 30 days. Two Na:Ca ratios of 18:1 and 1:2 were used for salinization of the nutrient solution. Both leaf dry weight and area decreased with increasing salinity, but in the more salinity tolerant cultivar H69-8235, the decrease was moderate. Salinity hardly reduced average area per leaf in H69-8235, while the number of leaves declined sharply. This decline was caused by enhanced senescence of mature leaves and not by a decreased rate of leaf initiation. In the more sensitive cultivar, H65-7052, leaf area and initiation of new leaves were sharply reduced by salinity while leaf senescence was less affected. Leaf water potential decreased during the early stages of salinity exposure, and the reduction in water potential was larger in H69-8235. Salinity also decreased the rate of transpiration rate but to a lesser extent than leaf development and growth. The accumulation of Cl and Na in the TVD (top visible dewlap) leaf of the tolerant cultivar H69-8235 was greater than in the sensitive cultivar H65-7052. The concentration of Cl in the TVD leaf was more than 10 times that of Na in both cultivars. The concentration of both ions, but not of K, increased during the early stages of salinity exposure and then remained constant. A gradient in concentration of Cl and Na over the plant was found in both cultivars at all salinity levels, and was steepest between the TVD and younger leaves. No specific Na effect on leaf growth or transpiration could be detected. The accumulation of Cl and Na but not of K occurred primarily in the roots rather than in the leaves and stalks. This revised version was published online in August 2006 with corrections to the Cover Date.     

不同CO2浓度和N肥水平对粳稻茎鞘非结构性碳水化合物含量与积累的影响

为了解CO2浓度升高和N肥水平对水稻茎鞘内非结构性碳水化合物(NSC)含量和积累量的影响,利用开顶式气室(OTC),以常规粳稻"南粳9108"为试验材料,设置3个CO2浓度水平:对照T0(背景大气)、T0+120μmol·mol-1(T1)和T0+200μmol·mol-1(T2)。在OTC内采用盆栽方式,设置3个氮(N)肥水平:10 g N·m^-2(N1)、20 g N·m^-2(N2)和30g N·m^-2(N3)。分别于水稻抽穗期、灌浆期(抽穗后20 d)和成熟期对地上部分各器官生物量、茎鞘NSC含量以及顶部四张叶片的N含量进行分析。结果表明:CO2浓度升高对抽穗期叶N含量总体无显著影响,但显著降低灌浆期N2和N3水平的叶N含量;CO2浓度升高对抽穗期茎鞘NSC含量和积累量无显著影响,抽穗期置换到高CO2浓度环境使灌浆期茎鞘NSC积累显著增加,置换到低CO2浓度环境使NSC积累显著减少。同一CO2浓度条件下,NSC含量和积累量均为N1>N2>N3,且N1处理均显著高于N3处理,CO2浓度升高和N水平的交互作用对灌浆期茎鞘NSC含量影响显著。水稻产量在不同CO2浓度水平间无显著差异,但随施氮水平的提高而增加。抽穗期与灌浆期水稻茎鞘NSC含量和积累量与茎鞘干重呈极显著正相关,与叶N含量呈极显著负相关;叶N衰减越慢,灌浆期水稻茎鞘NSC残留比(RNSC)越低;结实率和产量与RNSC呈显著负相关,RNSC越大,茎鞘NSC转移的越少,结实率和产量越低。     

Effect of salinity on growth,water use and nutrient use in radish (Raphanus sativus L.)

Radish (Raphanus sativus L.) plants were grown at five soil salinity levels (1, 2, 4, 9 and 13 dS m^-1) to analyse the effects on growth, dry matter partitioning, leaf expansion and water and nutrient use. Salinity was varied by proportionally changing the concentration of all macro nutrients. When the electrical conductivity (EC) of the soil solution increased from 1 to 13 dS m^-1, the influx concentration of the nutrients absorbed by the plants (the ratio between the uptakes of nutrients and water) increased only from 1.6 to 3.5 dS m^-1. The total nutrient uptake showed an optimum at an EC of the soil solution of about 4 dS m^-1. The data suggest that at low salinity level (≤ 2 dS m^-1) the nutrient uptake was limited by availability while at high salinity (>4 dS m^-1) it was limited by the growth of the plant. Total water use by the plants decreased and water use efficiency increased at high salinity. Plant growth was optimal at 2–4 dS m^-1. At salinities higher than 4 dS m^-1 total plant dry weight decreased 2.8% per dS m^-1. About 80% of the growth reduction at high salinity could be attributed to reduction of leaf area expansion and hence to reduction of light interception. The remaining 20% of the salinity effect on growth was most likely explained by a decrease in stomatal conductance. The small leaf area at high salinity was related to a reduced specific leaf area and increased tuber/shoot weight ratio. The latter could be attributed to tuber formation starting at a smaller plant size at high salinity. This revised version was published online in June 2006 with corrections to the Cover Date.     

Effects of Elevated Atmospheric CO2 Concentration on Leaf Anatomy and Morphology in Panicum Species Representing Different Photosynthetic Modes

Panicum tricanthum Nees, Panicum antidotale Retz., and Panicum decipiens Nees ex Trin. were selected to represent C3, C4, and C3/C4 intermediate perennial species of Panicum, respectively. Plants grown from seed with 900 ppm [CO2] under natural sunlight and controlled temperatures (30 degrees /22 degrees C) were compared with plants grown with ambient [CO2]. The anatomy of the last fully expanded leaf of the main tiller was studied by light microscopy with computerized graphic image analysis and by transmission electron microscopy. Leaf anatomy did not change qualitatively in response to elevated [CO2], but there were changes in leaf thickness and in the proportions of total transsectional area occupied by mesophyll, bundle sheath cells, vascular elements, and sclerenchyma, according to species. The abaxial stomatal frequency decreased by 22% for P. tricanthum but increased by ca. 30% for the other two species. With 900 ppm CO2, all three species showed a considerable increase in leaf starch content (to >30% of dry matter). Starch granules accumulated in chloroplasts of the mesophyll and bundle sheath cells. Increased leaf glaucousness in response to elevated [CO2] was the result of increased or modified deposition of epicuticular wax on both leaf surfaces, a response to elevated [CO2] that is unusual and one that has not been previously recorded for monocotyledons. The wax patterns were studied by scanning electron microscopy. Panicum decipiens did not respond to elevated [CO2] in a truly intermediate fashion; its responses resembled those of either the C3 or the C4 species. C3/C4 intermediates may thus be interpreted as developmental chimeras and not as species in transition between C3 and C4 modes in an evolutionary sense.     

Synthesis of Phytochelatins and Homo-Phytochelatins in Pisum sativum L

In the roots of pea plants (Pisum sativum L.) cultivated with 20 [mu]M CdCl2 for 3 d, synthesis of phytochelatins [PCs or ([gamma]EC)nG, where [gamma]EC is [gamma]glutamylcysteine and G is glycine] and homophytochelatins [h-PCs, ([gamma]EC)n[beta]-alanine] is accompanied by a drastic decrease in glutathione (GSH) content, but an increase in homoglutathione (h-GSH) content. In contrast, the in vitro activity of GSH synthetase increases 5-fold, whereas h-GSH synthetase activity increases regardless of Cd exposure. The consititutive enzyme PC synthase, which catalyzes the transfer of the [gamma]-EC moiety of GSH to an acceptor GSH molecule thus producing ([gamma]EC)2G, is activated by heavy metals, with Cd and Cu being strong activators and Zn being a very poor activator. Using h-GSH or hm-GSH for substrate, the synthesis rate of([gamma]EC)2[beta]-alanine and [gamma]EC)2-serine is only 2.4 and 0.3%, respectively, of the sythesis rate of ([gamma]EC)2G with GSH as substrate. However, in the presence of a constant GSH level, increasing the concentration of h-GSH or hm-GSH results in increased synthesis of ([gamma]EC)2[beta]-alanine or ([gamma]EC)2-serine, respecively; simultaneously, the synthesis of ([gamma]EC)2G is inhibited. [gamma]EC is not a substrate of PC synthase. These results are best explained by assuming that PC synthase has a [gamma]EC donor binding site, which is very specific for GSH, and a [gamma]EC acceptor binding site, which is less specific and accepts several tripeptides, namely GSH, h-GSH, and hm-GSH.     

Preliminary investigations on inducing salt tolerance in maize through inoculation with rhizobacteria containing ACC deaminase activity

Twenty rhizobacterial strains containing 1-aminocyclopropane-1-carboxylate deaminase were isolated from the rhizosphere of salt-affected maize fields. They were screened for their growth-promoting activities under axenic conditions at 1, 4, 8, and 12 dS x m-1 salinity levels. Based upon the data of the axenic study, the 6 most effective strains were selected to conduct pot trials in the wire house. Besides one original salinity level (1.6 dS x m-1), 3 other salinity levels (4, 8, and 12 dS x m-1) were maintained in pots and maize seeds inoculated with selected strains of plant growth-promoting rhizobacteria, as well as uninoculated controls were sown. Results showed that the increase in salinity level decreased the growth of maize seedlings. However, inoculation with rhizobacterial strains reduced this depression effect and improved the growth and yield at all the salinity levels tested. Selected strains significantly increased plant height, root length, total biomass, cob mass, and grain yield up to 82%, 93%, 51%, 40%, and 50%, respectively, over respective uninoculated controls at the electrical conductivity of 12 dS x m-1. Among various plant growth-promoting rhizobacterial strains, S5 (Pseudomonas syringae), S14 (Enterobacter aerogenes), and S20 (Pseudomonas fluorescens) were the most effective strains for promoting the growth and yield of maize, even at high salt stress. The relatively better salt tolerance of inoculated plants was associated with a high K+/Na+ ratio as well as high relative water and chlorophyll and low proline contents.     

Lateral CO2 diffusion inside dicotyledonous leaves can be substantial: quantification in different light intensities

Substantial lateral CO(2) diffusion rates into leaf areas where stomata were blocked by grease patches were quantified by gas exchange and chlorophyll a fluorescence imaging in different species across the full range of photosynthetic photon flux densities (PPFD). The lateral CO(2) flux rate over short distances was substantial and very similar in five dicotyledonous species with different vascular anatomies (two species with bundle sheath extensions, sunflower [Helianthus annuus] and dwarf bean [Phaseolus vulgaris]; and three species without bundle sheath extensions, faba bean [Vicia faba], petunia [Petunia hybrida], and tobacco [Nicotiana tabacum]). Only in the monocot maize (Zea mays) was there little or no evident lateral CO(2) flux. Lateral diffusion rates were low when PPFD <300 micromol m(-2) s(-1) but approached saturation in moderate PPFD (300 micromol m(-2) s(-1)) when lateral CO(2) diffusion represented 15% to 24% of the normal CO(2) assimilation rate. Smaller patches and higher ambient CO(2) concentration increased lateral CO(2) diffusion rates. Calculations with a two-dimensional diffusion model supported these observations that lateral CO(2) diffusion over short distances inside dicotyledonous leaves can be important to photosynthesis. The results emphasize that supply of CO(2) from nearby stomata usually dominates assimilation, but that lateral supply over distances up to approximately 1 mm can be important if stomata are blocked, particularly when assimilation rate is low.     

Increased Salt and Drought Tolerance by D-Ononitol Production in Transgenic Nicotiana tabacum L

A cDNA encoding myo-inositol O-methyltransferase (IMT1) has been transferred into Nicotiana tabacum cultivar SR1. During drought and salt stress, transformants (I5A) accumulated the methylated inositol D-ononitol in amounts exceeding 35 [mu]mol g-1 fresh weight In I5A, photosynthetic CO2 fixation was inhibited less during salt stress and drought, and the plants recovered faster than wild type. One day after rewatering drought-stressed plants, I5A photosynthesis had recovered 75% versus 57% recovery with cultivar SR1 plants. After 2.5 weeks of 250 mM NaCl in hydroponic solution, I5A fixed 4.9 [plus or minus] 1.4 [mu]mol CO2 m-2 s-1, whereas SR1 fixed 2.5 [plus or minus] 0.6 [mu]mol CO2 m-2 s-1. myo-Inositol, the substrate for IMT1, increases in tobacco under stress. Preconditioning of I5A plants in 50 mM NaCl increased D-ononitol amounts and resulted in increased protection when the plants were stressed subsequently with 150 mM NaCl. Pro, Suc, Fru, and Glc showed substantial diurnal fluctuations in amounts, but D-ononitol did not. Plant transformation resulting in stress-inducible, stable solute accumulation appears to provide better protection under drought and salt-stress conditions than strategies using osmotic adjustment by metabolites that are constitutively present.     

Alleviation of salinity stress in chickpea byRhizobium inoculation or nitrate supply

Influence of inoculation with efficient rhizobia or nitrate fertilization in alleviating salinity (NaCl, CaCl₂ and Na₂SO₄) stress was investigated in sand culture experiments. Shoot dry mass declined beyond salinity level corresponding to electrical conductivity (EC) 5.6 dS m^?1 in control or in inoculated plants and after EC 7.4 dS m^?1 in nitrate fed ones. Root growth was more sensitive and decreased at EC 3.3 dS m^?1. Nitrate reductase activity in leaves reduced at EC 3.3 dS m^?1 but in inoculated and nitrate fed plants it reduced at EC 5.6 dS m^?1. Na⁺ accumulation increased at EC 5.6 and 7.4 dS m^?1 in roots and, shoots, respectively. In inoculated and nitrate fed plants Na⁺ content in roots increased at EC 7.4 dS m^?1. Content of Ca²⁺ increased slightly only in shoots and content of K⁺ was unaffected. Besides inoculation, application of small doses of nitrogen should prove beneficial for legume cultivation in saline soils.     

Predominant localization of mitochondria enriched with glycine-decarboxylating enzymes in bundle sheath cells of Alternanthera tenella, a C3–C4 intermediate species

Mesophyll protoplasts and bundle sheath cells were prepared by enzymatic digestion of leaves of Alternanthera tenella, a C₃-C₄ intermediate species. The intercellular distribution of selected photosynthetic, photorespiratory and respiratory (mitochondrial) enzymes in these meso-phyll and bundle sheath cells was studied. The activity levels of photosynthetic enzymes such as PEP carboxylase (EC 4.1.1.31) or NAD-malic enzyme (EC 1.1.1.39) and photorespiratory enzymes such as glycolate oxidase (EC 1.1.3.1) or NADH-hydroxypyruvate reductase (EC 1.1.1.29) were similar in the two cell types. The activity levels of mitochondrial TCA cycle enzymes such as citrate synthase (EC 4.1.3.7) or fumarase (EC 4.2.1.2) were 2- to 3-fold higher in bundle sheath cells. On the other hand, the activity levels of mitochondrial photorespiratory enzymes, namely glycine decarboxylase (EC 2.1.2.10) and serine hydroxymethyltransferase (EC 2.1.2.1), were 6-9-fold higher in bundle sheath cells than in mesophyll protoplasts. Such preferential localization of mitochondria enriched with the glycine-decarboxylating system in the inner bundle sheath cells would result in efficient refixa-tion of CO₂ from not only photorespiration but also dark respiration before its exit from the leaf. We propose that predominant localization of mitochondria specialized in glycine decarboxylation in bundle sheath cells may form the basis of reduced photorespiration in this C₃-C₄ intermediate species.