Effects of salt-tolerant rootstock grafting on ultrastructure,photosynthetic capacity,and H<Subscript>2</Subscript>O<Subscript>2</Subscript>-scavenging system in chloroplasts of cucumber seedlings under NaCl stress

Plant growth, photosynthetic parameters, chloroplast ultrastructure, and the ascorbate-glutathione cycle system in chloroplasts of self-grafted and rootstock-grafted cucumber leaves were investigated. Grafted plants were grown hydroponically and were exposed to 0, 50, and 100 mM NaCl concentrations for 10 days. Under NaCl stress, the hydrogen peroxide (H₂O₂) content in cucumber chloroplasts increased, the chloroplast ultrastructure was damaged, and the gas stomatal conductance, intercellular CO₂ concentration, as well as shoot dry weight, plant height, stem diameter, leaf area, and leaf relative water content were inhibited, whereas these changes were less severe in rootstock-grafted plants. The activities of ascorbate peroxidase (APX; EC 1.11.1.11), glutathione reductase (GR; EC 1.6.4.2), and dehydroascorbate reductase (DHAR EC 1.8.5.1) were higher in the chloroplasts of rootstock-grafted plants compared with those of self-grafted plants under 50 and 100 mM NaCl. Similar trends were shown in leaf net CO₂ assimilation rate and transpiration rate, as well as reduced glutathione content under 100 mM NaCl. Results suggest that rootstock grafting enhances the H₂O₂-scavenging capacity of the ascorbate–glutathione cycle in cucumber chloroplasts under NaCl stress, thereby protecting the chloroplast structure and improving the photosynthetic performance of cucumber leaves. As a result, cucumber growth is promoted.     

Effects of elevated CO<Subscript>2</Subscript> and/or O<Subscript>3</Subscript> on hormone IAA in needles of Chinese pine

This study examined the effects of season-long exposure of Chinese pine (Pinus tabulaeformis) to elevated carbon dioxide (CO₂) and/or ozone (O₃) on indole-3-acetic acid (IAA) content, activities of IAA oxidase (IAAO) and peroxidase (POD) in needles. Trees grown in open-top chambers (OTC) were exposed to control (ambient O₃, 55 nmol mol⁻¹ + ambient CO₂, 350 μmol mol⁻¹, CK), elevated CO₂ (ambient O₃ + high CO₂, 700 μmol mol⁻¹, EC) and elevated O₃ (high O₃, 80 ± 8 nmol mol⁻¹ + ambient CO₂, EO) OTCs from 1 June to 30 September. Plants grown in elevated CO₂ OTC had a growth increase of axial shoot and needle length, compared to control, by 20% and 10% respectively, while the growth in elevated O₃ OTC was 43% and 7% less respectively, than control. An increase in IAA content and POD activity and decrease in IAAO activity were observed in trees exposed to elevated CO₂ concentration compared with control. Elevated O₃ decreased IAA content and had no significant effect on IAAO activity, but significantly increased POD activity. When trees pre-exposed to elevated CO₂ were transferred to elevated O₃ (EC–EO) or trees pre-exposed to elevated O₃ were transferred to elevated CO₂ (EO–EC), IAA content was lower while IAAO activity was higher than that transferred to CK (EC–CK or EO–CK), the change in IAA content was also related to IAAO activity. The results indicated that IAAO and POD activities in Chinese pine needles may be affected by the changes in the atmospheric environment, resulting in the change of IAA metabolism which in turn may cause changes in Chinese pine’s growth. An erratum to this article can be found at     

高浓度二氧化碳和臭氧对蒙古栎叶片活性氧代谢的影响

利用开顶箱熏蒸法,研究了高浓度O₃(≈80 nmol·mol^-1)和高浓度CO₂(≈700 μmol·mol^-1)及其复合处理对蒙古栎叶片活性氧代谢的影响.结果表明：高浓度O₃显著增加了蒙古栎叶片超氧阴离子(O₂^•)产生速率、过氧化氢(H₂O₂)和丙二醛(MDA)含量和电解质外渗率(P<0.05),显著降低了超氧化物歧化酶(SOD)、过氧化氢酶(CAT)、抗坏血酸过氧化物酶(APX)活性和抗坏血酸(AsA)含量(P<0.05).高浓度CO₂对蒙古栎叶片活性氧代谢影响不显著.高浓度O₃和CO₂复合处理的叶片O₂^•产生速率、H₂O₂和MDA含量和电解质外渗率上升不明显,说明高浓度CO₂缓解了高浓度O₃对蒙古栎叶片的氧化胁迫.复合处理的叶片SOD、CAT、APX活性以及AsA和总酚含量显著高于O₃处理的叶片(P<0.05),说明高浓度CO₂缓解了高浓度O₃对叶片抗氧化系统的消极影响.     

Exogenous H<Subscript>2</Subscript>O<Subscript>2</Subscript> improves the chilling tolerance of manilagrass and mascarenegrass by activating the antioxidative system

The effect of foliar pretreatment by hydrogen peroxide (H₂O₂) at low concentrations of 0, 5, 10, and 15 mM on the chilling tolerance of two Zoysia cultivars, manilagrass (Zoysia matrella) and mascarenegrass (Zoysia tenuifolia), was studied. The optimal concentration for H₂O₂ pretreatment was 10 mM, as demonstrated by the lowest malondialdehyde (MDA) content and electrolyte leakage (EL) levels and higher protein content under chilling stress (7°C/2°C, day/night). Prior to initiation of chilling, exogenous 10 mM H₂O₂ significantly increased catalase (CAT), ascorbate peroxidase (APX), glutathione-dependent peroxidases (GPX), and glutathione-S-transferase (GST) activities in manilagrass, and guaiacol peroxidase (POD), APX, and glutathione reductase (GR) activities in mascarenegrass, suggesting that H₂O₂ may act as a signaling molecule, inducing protective metabolic responses against further oxidative damage due to chilling. Under further stress, optimal pretreatments alleviated the increase of H₂O₂ level and the decrease of turfgrass quality, and improved CAT, POD, APX, GR, and GPX activities, with especially significant enhancement of APX and GPX activities from the initiation to end of chilling. These antioxidative enzymes were likely the important factors for acquisition of tolerance to chilling stress in the two Zoysia cultivars. Our results showed that pretreatment with H₂O₂ at appropriate concentration may improve the tolerance of warm-season Zoysia grasses to chilling stress, and that manilagrass had better tolerance to chilling, as evaluated by lower MDA and EL, and better turfgrass quality, regardless of the pretreatment applied.     

Exogenous Salicylic Acid Alleviates Growth Inhibition and Oxidative Stress Induced by Hypoxia Stress in <Emphasis Type="Italic">Malus robusta</Emphasis> Rehd

Salicylic acid (SA) as a signal molecule mediates many biotic and environmental stress-induced physiologic responses in plants. In this study we investigated the role of SA in regulating growth and oxidative stress in Malus robusta Rehd under both normoxic and hypoxic conditions. Hypoxia stress inhibited plant growth and dramatically reduced biomass. Addition of SA significantly alleviated the plant growth inhibition. The amounts of superoxide radicals (O₂ ⁻) and hydrogen peroxide (H₂O₂) significantly increased in leaves of the plants exposed to hypoxia stress and resulted in oxidative stress, which was indicated by accumulated concentration of malondialdehyde (MDA) and electrolyte leakage. Addition of SA significantly decreased the level of O₂ ⁻, electrolyte leakage, and lipid peroxidation and enhanced the activities of superoxide dismutase (SOD), peroxidase (POD), and ascorbate peroxidase (APX) under hypoxia stress. As important antioxidants, ascorbate (AsA) and glutathione (GSH) contents in the plant leaves were slightly increased by SA treatment compared to hypoxia stress treatment alone. It was concluded that SA could alleviate the detrimental effects of hypoxia stress on plant growth and of oxidative stress by enhancing the antioxidant defense system in leaves of M. robusta Rehd.     

Arsenite treatment induces oxidative stress,upregulates antioxidant system,and causes phytochelatin synthesis in rice seedlings

The effects of arsenite treatment on generation of reactive oxygen species, induction of oxidative stress, response of antioxidative system, and synthesis of phytochelatins were investigated in two indica rice (Oryza sativa L.) cvs. Malviya-36 and Pant-12 grown in sand cultures for a period of 5–20 days. Arsenite (As₂O₃; 25 and 50 μM) treatment resulted in increased formation of superoxide anion (O₂^.−), elevated levels of H₂O₂ and thiobarbituric acid reactive substances, showing enhanced lipid peroxidation. An enhanced level of ascorbate (AA) and glutathione (GSH) was observed irrespective of the variation in the level of dehydroascorbate (DHA) and oxidized glutathione (GSSG) which in turn influenced redox ratios AA/DHA and GSH/GSSG. With progressive arsenite treatment, synthesis of total acid soluble thiols and phytochelatins (PC) increased in the seedlings. Among antioxidative enzymes, the activities of superoxide dismutase (EC 1.15.1.1), catalase (EC 1.11.1.6), total ascorbate peroxidase (APX, EC 1.11.1.11), chloroplastic ascorbate peroxidase, guaiacol peroxidase (EC 1.11.1.7), monodehydroascorbate reductase (EC 1.6.5.4), and glutathione reductase (EC 1.6.4.2) increased in arsenite treated seedlings, while dehyroascorbate reductase (EC 1.8.5.1) activity declined initially during 5–10 days and increased thereafter. Results suggest that arsenite treatment causes oxidative stress in rice seedlings, increases the levels of many enzymatic and non-enzymatic antioxidants, and induces synthesis of thiols and PCs, which may serve as important components in mitigating arsenite-induced oxidative damage.     

Enhancement of antioxidant production in <Emphasis Type="Italic">Spirulina platensis</Emphasis> under oxidative stress

The present study examined the possibility of increasing the contents of some bioactive compounds of Spirulina platensis cultivated in medium containing various hydrogen peroxide concentrations (2, 4, 6 and 8 mM) as a model for environmental stress. A positive correlation was observed between the increase of H₂O₂ and increasing amounts of cellular lipophilic antioxidants (total carotenoids and α-tocopherol) and hydrophilic antioxidants [glutathione (GSH) and ascorbic acid (AsA)]. HPLC profile of carotenoids revealed that algae responded to the change of H₂O₂ exposure by the accumulation of higher amounts of β-carotene, astaxanthine, luteine, zeaxanthin and cryptoxanthin. S. platensis showed significant linear increase in activities of antioxidant enzymes, i.e., catalase (CAT), peroxidase (PX), ascorbate peroxidase (APX) and superoxide dismutase (SOD), with increasing H₂O₂ concentrations. A pronounced increase of oxidative lesions’ indexes [thiobarbituric acid reactive substances (TBARS) and paramagnetic radical-EPR signal] was found in algal grown at 8 mM H₂O₂. These data revealed that S. platensis behaved with different strategies against H₂O₂ exposure which is dose dependent and their response strongly correlated with the scavenging enzymes (SOD, CAT, PX and APX) and antioxidant compounds (GSH, AsA, β-carotene, astaxanthine and α-tocopherol) in the antioxidant defense systems. Therefore, S. platensis could be considered as good candidates for successful cultivation in artificial open ponds under different environmental conditions, as high value health foods, functional foods and as source of wide spectrum of nutrients.     

Plant growth and responses of antioxidants of <Emphasis Type="Italic">Chenopodium album</Emphasis> to long-term NaCl and KCl stress

The effects of long-term NaCl and KCl treatment on plant growth and antioxidative responses were investigated in Chenopodium album, a salt-resistant species widely distributed in semi-arid and light-saline areas of Xinjiang, China. Growth parameters [plant height, branch number, leaf morphology and chlorophyll (Chl) content], the level of oxidative stress [superoxide anion radical (O₂ ⁻), hydrogen peroxide (H₂O₂) and malondialdehyde (MDA) concentrations], activity of antioxidant enzymes [superoxide dismutase (SOD), catalase (CAT), peroxidase (POX)], the contents of non-enzymatic antioxidants [carotenoids (Car) and ascorbic acid (AsA)] and expression of selected genes were investigated. Plants were grown in the presence of 0, 50, and 300 mM NaCl or KCl for 2 months. Growth was stimulated by 50 mM NaCl or KCl, maintained stable at 300 mM NaCl, but was inhibited by 300 mM KCl. Three hundred mM NaCl did not affect O₂ ⁻, H₂O₂, MDA, Car and AsA, but increased the activities of SOD, CAT and POX compared to the controls. RT-PCR analysis suggested that expression of some genes encoding antioxidant enzymes could be induced during long-term salt stress, which was consistent with the enzyme activities. Treatment with 300 mM KCl was associated with elevated oxidative stress, and significantly decreased Car and AsA contents. These results suggest that an efficient antioxidant machinery is important for overcoming oxidative stress induced by treatment with high NaCl concentrations in C. album. Other strategies of ion regulation may also contribute to the differential tolerance to Na and K at higher concentrations.     

Responses of the antioxidative enzymes in Malaysian rice (<Emphasis Type="Italic">Oryza sativa</Emphasis> L.) cultivars under submergence condition

The potential involvement of activated oxygen species by submergence stress was studied in two Malaysian rice cultivars, MR219-4 and MR219-9, and cultivar FR13A that is known to be tolerant to submergence. Seedlings of these three rice cultivars were subjected to different submergence periods (4, 8, and 12 days). Under 8 days of complete submergence, FR13A cultivar showed higher lipid peroxidation in terms of malondialdehyde level and activities of antioxidative enzymes, superoxide dismutase (SOD), catalase (CAT), ascorbate peroxidase (APX), and glutathione reductase (GR) when compared to the MR219-4 and MR219-9 cultivars. MR219-9 showed higher SOD, APX, and GR activities after 12 days of submergence. The levels of SOD activity indicated that detoxification of O₂^·− to H₂O₂ was maintained at a stable level throughout the submergence stress until up to 8 days and increased rapidly at 12 days of submergence. The results indicated that tolerance to submergence in rice is associated until 8 days submergence for MR219-4 and FR13A cultivars. These findings suggested that tolerance to submergence stress in rice might be proven by increased the capacity of antioxidative system. In addition, CAT activity has much higher affinity for scavenges H₂O₂ than APX. Therefore, ascorbate glutathione cycle might be more efficient to scavenge H₂O₂.     

Effects of Ca(NO<Subscript>3</Subscript>)<Subscript>2</Subscript> stress on oxidative damage,antioxidant enzymes activities and polyamine contents in roots of grafted and non-grafted tomato plants

The effects of Ca(NO₃)₂ stress on biomass production, oxidative damage, antioxidant enzymes activities and polyamine contents in roots of grafted and non-grafted tomato plants were investigated. Results showed that when exposed to 80 mM Ca(NO₃)₂ stress, the biomass production reduction in non-grafted plants was more significant than that of grafted plants. Under Ca(NO₃)₂ stress, superoxide anion radical (O₂•⁻) producing rate, hydrogen peroxide (H₂O₂) and malondialdehyde (MDA) contents of non-grafted plants roots were significantly higher than those of grafted plants, however, nitrate (NO₃ ⁻), ammonium (NH₄ ⁺) and proline contents, superoxide dismutase (SOD, EC1.15.1.1), peroxidase (POD, EC1.11.1.7), catalase (CAT, EC1.11.1.6) and arginine decarboxylase (ADC, EC 4.1.1.19) activities of grafted plants roots were significantly higher than those of non-grafted plants. Regardless of stress, free, conjugated and bound polyamine contents in roots of grafted plants were significantly higher than those of non-grafted plants. The possible roles of antioxidant enzymes, prolines and polyamines in adaptive mechanism of tomato roots to Ca(NO₃)₂ stress were discussed. Gu-Wen Zhang and Zheng-Lu Liu contributed equally to this work.     

Drought-induced oxidative damage and antioxidant responses in peanut (<Emphasis Type="Italic">Arachis</Emphasis><Emphasis Type="Italic">hypogaea</Emphasis> L.) seedlings

Two cultivars of peanut (Arachis hypogaea L.) which were designated as resistant (Florispan) and sensitive (Gazipasa) according to their growth retardation under drought stress conditions were compared for their oxidative damage and antioxidant responses. Sixteen days-old peanut seedlings were subjected to PEG-6000 solutions of two different osmotic potentials; −0.4 and −0.8 MPa, and various growth parameters, photosystem II activity, changes in malondialdehyde (MDA), hydrogen peroxide (H₂O₂) and proline levels, activities of ascorbate peroxidase (APX), catalase (CAT), peroxidase (POX) and gluthatione reductase (GR) enzymes were determined. Both cultivars exhibited water deficit at −0.8 MPa osmotic potential of PEG-6000 and H₂O₂ levels significantly increased during exposure to −0.4 MPa osmotic potential. However, H₂O₂ levels were under control in both cultivars at exposure to −0.8 MPa osmotic potential. Significant proline accumulation was observed in the tissues of cv. Florispan at −0.8 MPa osmotic potential, whereas proline accumulation did not appear to be an essential part of the protection mechanism against drought in cv. Gazipasa. No significant variation in chlorophyll fluorescence values were detected in neither of the cultivars. Enzyme activity measurements revealed that Gazipasa copes well with lesser magnitudes of drought stress by increasing the activity of mainly APX, and during harsh stress conditions, only APX maintains its activity in the tissues. In cultivar Florispan, GR activity appears to take role in lesser magnitudes of drought stress, whereas CAT and APX activities appear to be very crucial antioxidative defenses during intense stress conditions. The results indicate that, the level of proline and activities of the enzymes CAT and APX are important mechanisms for the maintenance of drought tolerance in peanut plants.     

Antioxidative response of <Emphasis Type="Italic">Hordeum maritimum</Emphasis> L<Emphasis Type="Italic">.</Emphasis> to potassium deficiency

The objective of the present study was to determine the influence of potassium deprivation on the halophyte species Hordeum maritimum grown in hydroponics for 2 weeks. Treatments were with potassium (+K) or without potassium (−K). Growth, water status, mineral nutrition, parameters of oxidative stress [malondialdehyde (MDA), carbonyl groups (C=O), and hydrogen peroxide concentration (H₂O₂) contents], antioxidant enzyme activities [superoxide dismutase (SOD, EC 1.15.1.1), catalase (CAT, EC 1.11.1.6), guaiacol peroxidase (GPX, EC 1.11.1.7), ascorbate peroxidase (APX, EC 1.11.1.11), monodehydroascorbate peroxidase (MDHAR, EC 1.6.5.4), dehydroascorbate peroxidase (DHAR, EC 1.8.5.1), and glutathione reductase (GR, EC 1.6.4.2)], and antioxidant molecules [ascorbate (ASC), and glutathione (GSH)] were determined. Results showed that the growth of vegetative organs decreased owing to potassium deficiency with roots (−36%) more affected than shoots (−12%). Water status was only diminished in roots (reduction of 24%). Potassium deprivation decreased potassium concentration in both organs, this decrease was more pronounced in roots (−81%) than in shoots (−55%). In contrast to carbonyl groups, MDA content increased owing to potassium deprivation. Except for CAT activity that remained unaffected; SOD, GPX, APX, GR, MDHAR, and DHAR activities were significantly increased. H₂O₂ concentration was negatively correlated with the activities of enzymes and the accumulation of non-enzymatic antioxidants implicated in its detoxification. In conclusion, a cooperative process between the antioxidant systems is important for the tolerance of H. maritimum to potassium deficiency.     

Cadmium effects on mineral nutrition and stress in <Emphasis Type="Italic">Potamogeton crispus</Emphasis>

The mechanisms of plant tolerance to cadmium stress were studied by short-term exposure of Potamogeton crispus L. to various concentrations of Cd ranging from 0 to 0.09 mM. The accumulation of Cd and its influence on nutrient elements, chlorophyll pigments, ultrastructure, proline and MDA contents, and free radical production, as well as the activities of superoxide dismutase (SOD), peroxidase (POD), ascorbate peroxidase (APX), and glutathione reductase (GR) were investigated. The higher Cd concentration in the medium resulted in a significant enhancement of Cd accumulation. Photosynthetic pigment content decreased and ultrastructural damage to the leaf cells was aggravated with the increase in the Cd concentrations. Disruption of chloroplasts and mitochondria was observed even at the lowest concentration of Cd. Meantime, the rate of O₂^*− generation and the contents of H₂O₂ and MDA significantly increased under Cd stress, suggesting that Cd caused oxidative stress. In addition, the antioxidant system was clearly activated following Cd exposure. SOD and POD activities increased initially and then decreased, while APX and GR activities markedly increased. Simultaneously, mineral nutrition was disturbed. While K, P, Ca, and Cu contents decreased, Na, Fe, and Mn contents increased. Induction of antioxidant enzyme activities in leaves exposed to elevated Cd concentrations may be involved in Cd tolerance of P. crispus.     

The Effects of Exogenous Ascorbic Acid on the Mechanism of Physiological and Biochemical Responses to Nitrate Uptake in Two Rice Cultivars (<Emphasis Type="Italic">Oryza sativa</Emphasis> L.) Under Aluminum Stress

As a major antioxidant in plants, ascorbic acid (AsA) plays a very important role in the response to aluminum (Al) stress. However, the effect of AsA on the mitigation of Al toxicity and the mechanism of nitrate nitrogen (NO₃ ^?–N) uptake by plants under Al stress are unclear. In this study, a hydroponic experiment was conducted using peak 1 A rice (sterile line, Indica) with weaker resistance to Al and peak 1 superior 5 rice (F1 hybrid, Indica) with stronger resistance to Al to study the effects of exogenous AsA on the physiological and biochemical responses to NO₃ ^?–N uptake by rice roots exposed to 50 μmol L^?1 Al. Al stress induced increases in the concentrations of H₂O₂ and malondialdehyde (MDA) and in the activities of antioxidant enzymes [such as superoxide dismutase (SOD), peroxidase (POD), catalase (CAT), and ascorbate peroxidase (APX)]. Plasma membrane (PM) H⁺-ATPase and H⁺-pump activities, endogenous AsA content and NO₃ ^?–N uptake in rice roots decreased under Al stress. After treatment with 2 mmol L^?1 exogenous AsA combined with Al, concentrations of H₂O₂ and MDA in roots notably decreased, and endogenous AsA content and activities of SOD, POD, CAT, and APX in rice roots increased significantly; furthermore, the interaction of PM H⁺-ATPase and the 14-3-3 protein was also enhanced significantly compared with that in control plants without AsA treatment, which clearly increased NO₃ ^?–N uptake. Based on all of these data, the application of AsA significantly reduced the accumulation of H₂O₂ and MDA and increased the activities of PM H⁺-ATPase and the H⁺-pump by increasing the endogenous AsA content, the antioxidant enzyme activities, and the interaction of PM H⁺-ATPase and the 14-3-3 protein in the roots of the two rice cultivars under Al stress, thereby improving the uptake of NO₃ ^?–N in rice.     

Manganese-excess induces oxidative stress,lowers the pool of antioxidants and elevates activities of key antioxidative enzymes in rice seedlings

Manganese (Mn) is an essential element for plant growth but in excess, specially in acidic soils, it can become phytotoxic. In order to investigate whether oxidative stress is associated with the expression of Mn toxicity during early seedling establishment of rice plants, we examined the changes in the level of reactive oxygen species (ROS), oxidative stress induced an alteration in the level of non-enzymic antioxidants and activities of antioxidative enzymes in rice seedlings grown in sand cultures containing 3 and 6 mM MnCl₂. Mn treatment inhibited growth of rice seedlings, the metal increasingly accumulated in roots and shoots and caused damage to membranes. Mn treated plants showed increased generation of superoxide anion (O₂ ^.−), elevated levels of H₂O₂ and thiobarbituric acid reactive substances (TBARS) and decline in protein thiol. The level of nonprotein thiol, however, increased due to Mn treatment. A decline in contents of reduced ascorbate (AsA) and glutathione (GSH) as well as decline in ratios of their reduced to oxidize forms was observed in Mn-treated seedlings. The activities of antioxidative enzymes superoxide dismutase (SOD) and its isoforms Mn SOD, Cu/Zn SOD, Fe SOD as well as guaiacol peroxidase (GPX) increased in the seedlings due to Mn treatment however, catalase (CAT) activity increased in 10 days old seedlings but it declined by 20 days under Mn treatment. The enzymes of Halliwell-Asada cycle, ascorbate peroxidase (APX) monodehydoascorbate reductase (MDHAR), dehyroascorbate reductase (DHAR) and glutathione reductase (GR) increased significantly in Mn treated seedlings over controls. Results suggest that in rice seedlings excess Mn induces oxidative stress, imbalances the levels of antioxidants and the antioxidative enzymes SOD, GPX, APX and GR appear to play an important role in scavenging ROS and withstanding oxidative stress induced by Mn.     

Interactive effects of gibberellin A<Subscript>3</Subscript> and ascorbic acid on lipid peroxidation and antioxidant enzyme activities in <Emphasis Type="Italic">Glycine max</Emphasis> seedlings under nickel stress

The effects of nickel in combination with ascorbic acid (AsA) and gibberellin on 7-day-old soybean seedlings were examined. Exposure to 0.25 mM NiCl₂ × 6H₂O for 5 days resulted in toxicity symptoms, such as formation of reddish-brown mottled spots on the leaf blade. Addition of 0.05 mM GA₃ or 1 mM AsA reduced toxic effects of nickel. After their simultaneous application, these symptoms did not appear. Ni decreased dry weights of roots and shoots and reduced chlorophyll content in leaves. An enhanced level of lipoxygenase activity and malondialdehyde, and changes in the activities of the antioxidant enzymes, catalase, guaiacol peroxidase and ascorbate peroxidase, in both roots and leaves indicated that Ni caused an oxidative stress in soybean plants. The Ni-stressed seedlings exposed to AsA or GA₃, especially to GA₃ plus AsA, exhibited an improved growth as compared to Ni-treated plants. GA₃ decreased Ni uptake by roots, while ascorbic acid considerably reduced root-to-shoot translocation of Ni. Interaction of AsA plus GA₃ prevented the decrease in chlorophyll content and lipid peroxidation as well as increased the activities of the antioxidant enzymes. These results suggest that GA₃ plus AsA treatment counteracts the negative effects of Ni on soybean seedlings. Published in Russian in Fiziologiya Rastenii, 2007, Vol. 54, No. 1, pp. 85–91. The text was submitted by the authors in English.     

Variations of vinblastine accumulation and redox state affected by exogenous H<Subscript>2</Subscript>O<Subscript>2</Subscript> in <Emphasis Type="Italic">Catharanthus roseus</Emphasis> (L.) G. Don

Effects of exogenous H₂O₂ application on vinblastine (VBL) and its precursors, vindoline (VIN), catharanthine (CAT) and α-3′,4′-anhydrovinblastine (AVBL), were measured in Catharanthus roseus seedlings in order to explore possible correlation of VBL formation with oxidative stress. VBL accumulation has previously been shown to be regulated by an in vitro H₂O₂-dependent peroxidase (POD)-like synthase. Experimental exposure of plants to different concentrations of H₂O₂ showed that endogenous H₂O₂ and alkaloid concentrations in leaves were positively elevated. The time-course variations of alkaloid concentrations and redox state, reflected by the concentrations of H₂O₂, ascorbic acid (AA), oxidative product of glutathione (GSSG) and POD activity, were significantly altered due to H₂O₂ application. The further correlation analysis between alkaloids and redox status indicated that VBL production was tightly correlated with redox status. These results provide a new link between VBL metabolisms and redox state in C. roseus.     

Down-regulation of Rubisco activity under combined increases of CO<Subscript>2</Subscript> and temperature minimized by changes in Rubisco k<Subscript>cat</Subscript> in wheat

Increases in growth temperature have been observed to affect photosynthesis differently under long-term exposure to ambient- and twice ambient-air CO₂ concentrations. This study investigates the causes of this interaction in wheat (Triticum aestivum L.) grown in the field over two consecutive years under temperature gradient chambers in ambient (370 μmol mol⁻¹) or elevated (700 μmol mol⁻¹) atmospheric CO₂ concentrations and at ambient or ambient +4°C temperatures, with either a low or a high nitrogen supply. The photosynthesis-internal CO₂ response curves and the activity, activation state, k_cat and amount of Ribulose-1, 5-bisphosphate carboxylase/oxygenase (Rubisco) were measured, as well as the soluble protein concentration in flag leaves at ear emergence and 8–15 days after anthesis. A high nitrogen supply increased V_cmax, the Rubisco amount and activity and soluble protein contents, but did not significantly change the Rubisco k_cat. Both elevated CO₂ and above ambient temperatures had negative effects on V_cmax and Rubisco activity, but at elevated CO₂, an increase in temperature did not decrease V_cmax or Rubisco activity in relation to ambient temperature. The amounts of Rubisco and soluble protein decreased with elevated CO₂ and temperature. The negative impact of elevated CO₂ on Rubisco properties was somewhat counteracted at elevated temperatures by an increase in k_cat. This effect can diminish the detrimental effects on photosynthesis of combined increases of CO₂ and temperature.     

Relationship between leaf reddening,ROS and antioxidants in <Emphasis Type="Italic">Buxus microphylla</Emphasis> during overwintering

To provide a theoretical basis for revealing the mechanism of winter leaf reddening in evergreen species, the relationships between winter leaf reddening, reactive oxygen species (ROS) and the antioxidant system of Buxus microphylla ‘Wintergreen’ were studied. The pigment changes, ROS production, lipid peroxidation and antioxidants activities of sun leaves during the reddening and regreening processes were investigated, using green shade leaves as controls. The carotenoids in the sun leaves increased linearly with reddening but decreased with the regreening. There was no significant difference in either the superoxide anions (O ₂ ^?· ) or malondialdehyde (MDA) changes between the sun and shade leaves, and their O ₂ ^?· contents were positively correlated with MDA. In contrast to the shade leaves, the sun leaves showed a trend in which the hydrogen peroxide (H₂O₂) changes were closely related to the reddening process and positively correlated with carotenoids content but not with the MDA content. A similar trend was observed for catalase (CAT) and ascorbate peroxidase (APX) activity between the sun and shade leaves, but superoxide dismutase (SOD) and peroxidase (POD) activity and the ascorbate (AsA) content differed between these two ecotypes. Furthermore, the sun leaves had higher CAT activity and AsA content than the corresponding shade leaves. These results suggested that H₂O₂ might play an important role in the winter reddening of sun leaves by promoting the accumulation of carotenoids. In addition, SOD, POD and AsA probably play a photoprotective role in winter-red sun leaves, while the changes in O ₂ ^?· , CAT and APX were independent of winter leaf reddening and were more likely responses to stress caused by low temperatures.     

Growth and photosynthetic response of <Emphasis Type="Italic">Fagus crenata</Emphasis> seedlings to ozone and/or elevated carbon dioxide

We investigated the effects of ozone (O₃) and/or elevated CO₂ concentration ([CO₂]) on growth and photosynthetic traits of Fagus crenata seedlings. Two-year-old seedlings were grown in four experimental treatments comprising two O₃ treatments (charcoal-filtered air and 100 nmol mol⁻¹ O₃; 6 h/day, 3 days/week) in combination with two CO₂ treatments (350 and 700 μmol mol⁻¹) for 18 weeks in environmental control growth chambers. The four treatments were designated as control, elevated O₃, elevated CO₂, and elevated CO₂ + O₃. Dry matter growth of the seedlings was greater in elevated CO₂ + O₃ than in elevated CO₂. In elevated CO₂ + O₃, a marked increase of second-flush leaves, considered a compensative response to O₃, was observed. The net photosynthetic rate of first-flush leaves in elevated CO₂ + O₃ increased earlier and was maintained for a longer period of time than that in elevated CO₂. Because emergence of second-flush leaves of F. crenata is greatly affected by the amount of assimilation products of first-flush leaves in current year, we consider that an early increase in the net photosynthetic rate of first-flush leaves contributed to the marked increase in second-flush leaf emergence under elevated CO₂ + O₃. These results imply that we must account for changes in compensative capacity with respect to not only morphological traits but also phenological traits and physiological functions such as photosynthesis when evaluating effects of O₃ on F. crenata under elevated [CO₂].