Effect of abscisic acid on heat stress tolerance in the calli from two ecotypes of <Emphasis Type="Italic">Phragmites communis</Emphasis>

Dune reed (DR) and swamp reed (SR) are two ecotypes of reed (Phragmites communis Trin.) that displayed differences in stress tolerance. To uncover the molecular basis for such difference, the effects of heat stress were studied using the calli derived from the two ecotypes. Heat stress caused increased ion leakage, inhibited growth, decreased cell viability, and elevated hydrogen peroxide (H₂O₂) and malondialdehyde (MDA) contents in the calli of both ecotypes, but DR callus showed better heat tolerance than SR callus. In DR callus, heat stress caused significant increase in the endogenous ABA content but not in SR callus. Application of fluridone (an ABA synthesis inhibitor) aggravated the heat stress damages on the DR callus whereas it had only minimal impact on the SR callus. Exogenous application of ABA alleviated the heat stress symptoms in the calli of both ecotypes. ABA treatment increased the activities of superoxide dismutase, catalase, ascorbate peroxidase and peroxidase, and also decreased H₂O₂ and MDA contents. These results indicate that the ability of ABA synthesis under heat stress is a key factor attributing to the higher heat tolerance of DR than SR.     

Glucose-6-phosphate dehydrogenase plays a central role in modulating reduced glutathione levels in reed callus under salt stress

In the present study, we investigated the role of glucose-6-phosphate dehydrogenase (G6PDH) in regulating the levels of reduced form of glutathione (GSH) to the tolerance of calli from two reed ecotypes, Phragmites communis Trin. dune reed (DR) and swamp reed (SR), in a long-term salt stress. G6PDH activity was higher in SR callus than that of DR callus under 50–150 mM NaCl treatments. In contrast, at higher NaCl concentrations (300–600 mM), G6PDH activity was lower in SR callus. A similar profile was observed in GSH contents, glutathione reductase (GR) and glutathione peroxidase (GPX) activities in both salt-stressed calli. After G6PDH activity and expression were reduced in glycerol treatments, GSH contents and GR and GPX activity decreased strongly in both calli. Simultaneously, NaCl-induced hydrogen peroxide (H₂O₂) accumulation was also abolished. Exogenous application of H₂O₂ increased G6PDH, GR, and GPX activities and GSH contents in the control conditions and glycerol treatment. Diphenylene iodonium (DPI), a plasma membrane (PM) NADPH oxidase inhibitor, which counteracted NaCl-induced H₂O₂ accumulation, decreased these enzymes activities and GSH contents. Furthermore, exogenous application of H₂O₂ abolished the N-acetyl-l-cysteine (NAC)-induced decrease in G6PDH activity, and DPI suppressed the effect of buthionine sulfoximine (BSO) on induction of G6PDH activity. Western-blot analyses showed that G6PDH expression was stimulated by NaCl and H₂O₂, and blocked by DPI in DR callus. Taken together, G6PDH activity involved in GSH maintenance and H₂O₂ accumulation under salt stress. And H₂O₂ regulated G6PDH, GR, and GPX activities to maintain GSH levels. In the process, G6PDH plays a central role.     

Nitric oxide protects against polyethylene glycol-induced oxidative damage in two ecotypes of reed suspension cultures

Dune reed (DR) is the more tolerant ecotype of reed to environmental stresses than swamp reed (SR). Under osmotic stress mediated by polyethylene glycol (PEG-6000), the suspension culture of SR showed higher ion leakage, and more oxidative damage to the membrane lipids and proteins was observed compared with the relatively tolerant DR suspension culture. Treatment with sodium nitroprusside (SNP) can significantly alleviated PEG-induced ion leakage, thiobarbituric acid reactive substances (TBARS) and carbonyl contents increase in SR suspension culture. The levels of H(2)O(2) and O(2)(-) were reduced, and the activities of antioxidant enzymes such as superoxide dismutase (SOD), catalase (CAT) and ascorbate peroxidase (APX) were increased in both suspension cultures in the presence of SNP under osmotic stress, but lipoxygenase (LOX) activity was inhibited. 2-(4-carboxy-2-phenyl)-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide (PTIO), a specific Nitric oxide (NO) scavenger, blocked the SNP-mediated protection. Depletion of endogenous NO with PTIO strongly enhanced oxidative damage in DR compared with that of PEG treatment alone, whereas had no effect on SR. Moreover, NO production increased significantly in DR while kept stable in SR under osmotic stress. Taken together, these results suggest that PEG induced NO release in DR but not SR can effectively protect against oxidative damage and confer an increased tolerance to osmotic stress in DR suspension culture.     

Pathways of ROS homeostasis regulation in Mesembryanthemum crystallinum L. calli exhibiting differences in rhizogenesis

A comparison of the hydrogen peroxide (H₂O₂) content, proline and betacyanin concentration and activities of some antioxidant enzymes (catalase, superoxide dismutase, guaiacol and ascorbate peroxidases) was made in Mesembryanthemum crystallinum L. calli differing in rhizogenic potential. Callus was induced from hypocotyls of 10-day-old seedlings on a medium containing 1?mg?l^?1 2,4-dichlorophenoxyacetic acid and 0.2?mg?l^?1 kinetin, which was either supplemented with 40?mM NaCl (CIM-NaCl medium) or did not contain any salt (CIM medium). The callus obtained on CIM-NaCl was rhizogenic, whereas the callus induced on the medium without salt was non-rhizogenic throughout the culture. The rhizogenic callus differed from the non-rhizogenic callus in lower betacyanin and H₂O₂ content, but the rhizogenic callus displayed a higher proline level. The activity of H₂O₂ scavenging enzymes, such as catalase (CAT), ascorbate peroxidase (APX) and guaiacol peroxidase (POD), was markedly higher in the rhizogenic callus than in the non-rhizogenic callus, but the total activity of superoxide dismutase (SOD) was higher in the non-rhizogenic callus than in the rhizogenic callus. Aminotriazole (CAT inhibitor) and diethyldithiocarbamate (SOD inhibitor) were added solely to the CIM and CIM-NaCl media to manipulate the concentration of reactive oxygen species (ROS) in the cultured tissues. Both CAT and SOD inhibitors brought about an increase in H₂O₂ content in calli cultured on CIM-NaCl and the loss of rhizogenic potential. Conversely, the addition of inhibitors to the medium without salt led to a decrease in H₂O₂ content. This corresponded with a significant decrease in the endogenous concentration of betacyanins, but did not change the lack of rhizogenic ability.     

Photosynthetic Electron Transport,Photophosphorylation, and Antioxidants in Two Ecotypes of Reed (Phragmites Communis Trin.) from Different Habitats

We compared chloroplast photochemical properties and activities of some chloroplast-localised enzymes in two ecotypes of Phragmites communis, swamp reed (SR, C₃-like) and dune reed (DR, C₄-like) plants growing in the desert region of north-west China. Electron transport rates of whole electron transport chain and photosystem (PS) 2 were remarkably lower in DR chloroplasts. However, the electron transport rate for PS1 in DR chloroplasts was more than 90 % of the activity similar in the SR chloroplasts. Activities of Mg²⁺-ATPase and cyclic and non-cyclic photophosphorylations were higher in DR chloroplasts than in the SR ones. The activities of chloroplast superoxide dismutase (SOD) and ascorbate peroxidase (APX), both localised at or near the PS1 complex and serving to scavenge active oxygen around PS1, and the content of ascorbic acid, a special substrate of APX in chloroplast, were all higher in DR chloroplasts. Hence reed, a hydrophytic plant, when subjected to intense selection pressure in dune habitat, elevates its cyclic electron flow around PS1. In consequence, it provides extra ATP required by C₄ photosynthesis. Combined high activities of active oxygen scavenging components in DR chloroplasts might improve protection of photosynthetic apparatus, especially PS1, from the damage of reactive oxygen species. This offers new explanation of photosynthetic performance of plant adaptation to long-term natural drought habitat, which is different from those, subjected to the short-term stress treatment or even to the artificial field drought.     

Exogenous spermidine and spermine enhance cadmium tolerance of <Emphasis Type="Italic">Potamogeton malaianus</Emphasis>

In order to investigate the effects of spermidine (Spd) and spermine (Spm) on cadmium stress, the content of chlorophyll, hydrogen peroxide (H₂O₂), malondialdehyde (MDA), soluble protein and proline, the rate of O₂^·− generation, and activities of antioxidant enzymes (superoxide dismutase (SOD), peroxidase (POD), ascorbate peroxidase (APX), and glutathione reductase (GR)) in Potamogeton malaianus Miq. were measured. Exogenous application of Spd or Spm significantly enhanced the level of proline, retarded the loss of chlorophyll, enhanced photosynthesis, decreased the rate of O₂^·− generation and H₂O₂ content, and prevented Cd-induced lipid peroxidation. Spd and Spm also effectively maintained the balance of antioxidant enzyme activities under Cd stress; however, GR activity was found to increase only slightly in response to polyamines (PAs). The antioxidant systems, which were modified by PAs, were able to moderate the radical-scavenging system and to lessen in this way the oxidative stress. These results suggest that both Spd and Spm can enhance Cd tolerance of P. malaianus.     

Effects of exogenous spermidine on antioxidant system of tomato seedlings exposed to high temperature stress

The effects of foliar spraying with spermidine (Spd) on antioxidant system in tomato (Lycopersicon esculentum Mill.) seedlings were investigated under high temperature stress. The high temperature stress significantly inhibited plant growth and reduced chlorophyll (Chl) content. Application of exogenous 1 mM Spd alleviated the inhibition of growth induced by the high temperature stress. Malondialdehyde (MDA), hydrogen peroxide (H₂O₂) content and superoxide anion (O₂) generation rate were significantly increased by the high temperature stress, but Spd significantly reduced the accumulation of reactive oxygen species (ROS) and MDA content under the stress. The high temperature stress significantly decreased glutathione (GSH) content and activities of superoxide dismutase (SOD), peroxidase (POD), catalase (CAT), ascorbate peroxidase (APX), glutathione reductase (GR), monodehydroascorbate reductase (MDHAR) and dehydroascorbate reductase (DHAR), but increased contents of dehydroascorbic acid (DHA), ascorbic acid (AsA), and oxidized glutathione (GSSG) in tomato leaves. However, Spd significantly increased the activities of antioxidant enzymes, levels of antioxidants and endogenous polyamines in tomato leaves under the high temperature stress. In addition, to varying degrees, Spd regulated expression of MnSOD, POD, APX2, APX6, GR, MDHAR, DHAR1, and DHAR2 genes in tomato leaves exposed to the high temperature stress. These results suggest that Spd could change endogenous polyamine levels and alleviate the damage by oxidative stress enhancing the non-enzymatic and enzymatic antioxidant system and the related gene expression.     

Calcium and L-histidine effects on ascorbate-glutathione cycle components under nickel-induced oxidative stress in tomato plants

The effects of NiSO₄, calcium, and L-histidine (His) on the components of ascorbate-glutathione cycle, antioxidant enzymes and lipid peroxidation in a tomato cultivar Early Urbana Y was investigated. The activities of enzymes including catalase (CAT), guaiacol peroxidase (GPX), ascorbate peroxidase (APX), superoxide dismutase (SOD), glutathione reductase (GR), lipoxygenase (LOX), and phenylalanine ammonia lyase (PAL) were measured. In addition, the content of H₂O₂, ascorbate (ASC), dehydroascorbate (DHA), reduced glutathione (GSH), chlorophyll (Chl) a+b, carotenoids, proteins, malondialdehyde (MDA), membrane aldehydes, and electrolyte leakage (EL) were determined. Results suggest that the excess of Ni increased the content of H₂O₂, MDA, membrane aldehydes and proteins in roots as well as GPX, LOX, APX activities, and EL in leaves, whereas Ca and His ameliorated these effects. Moreover, decreasing leaf GSH and DHA content and GR activity were observed under the Ni stress, but these parameters were raised by Ca plus His treatment. However, no improvement in leaf protein, ASC, root GSH content, and activities of PAL and CAT were observed by using Ca or His under Ni stress.     

Solute levels and osmoregulatory enzyme activities in reed plants adapted to drought and saline habitats

The pattern of solute accumulation and the activities of key enzymes involved in metabolism of proline and betaine were investigated in three ecotypes of reed from different habitats: swamp reed (SR), dune reed (DR), and heavy salt meadow reed (HSR). The two terrestrial reed ecotypes, DR and HSR, exhibited a higher capacity for osmotic adjustment; they accumulated higher contents of K⁺ and Ca²⁺ in the leaves in comparison with SR. DR also had the highest soluble sugar content in its leaves. HSR has higher levels of Na⁺ in its root environment and this was reflected by considerable accumulation of Na⁺ in the HSR rhizome. However, the different zones of its leaf lamina (upper, middle and lower) did not exhibit increased levels of Na⁺, suggesting that HSR has the ability to accumulate Na⁺ in the rhizome to protect the shoots from excessive Na⁺ toxicity. DR and HSR had higher levels of proline and betaine in the leaves than did SR. This difference was consistent with the activities of the various biosynthetic enzymes: betaine aldehyde dehydrogenase (BADH), pyrroline-5-carboxylate reductase (P5CR) and ornithine--aminotransferase (OAT) were enhanced in DR and HSR as compared to SR, whereas proline oxidase (PO) activities were inhibited. These findings suggest that changes in the activities of enzymes involved in osmotregulation might play important roles in the adaptation of reed, a hydrophilic plant, to more extreme dune and saline habitats. The relative contributions of the various proline synthetic pathways are also discussed.     

Antioxidant defense system in <Emphasis Type="Italic">Phragmites communis</Emphasis> Trin. ecotypes

The antioxidant defense system in three ecotypes of reed (Phragmites communis Trin.), swamp reed (SR), dune reed (DR), and heavy salt meadow reed (HSMR), from northwest China were investigated. The HSMR possessed the highest ratio of ascorbate (ASC)/dehydroascorbate (DHA) and activities of superoxide dismutase (SOD) and catalase among the three reed ecotypes, whereas, the DR exhibited the highest ratio of glutathione/glutathione disulfide and activities of ASC peroxidase (APX) and DHA reductase. Malondialdehyde and hydrogen peroxide contents were highest in HSMR, intermediate in SR, and lowest in DR. In addition, different isoenzymes of glutathion reductase, APX, SOD and DHA were also observed in three reed ecotypes.     

Effects of cytokinin on callus proliferation associated with physiological and biochemical changes in <Emphasis Type="Italic">Vitis vinifera</Emphasis> L.

The essential role of 6-benzylaminopurine (BA) in plant tissue culture has been widely known; however, physiological and biochemical mechanisms behind BA requirement have not been fully understood yet. BA may have an important role on callus growth by regulating antioxidant enzyme activities and acting as an effective free radical scavenger. To test this hypothesis, the impact of exogenous BA concentrations on antioxidative system in Vitis vinifera L. cv. ‘Bogazkere’ callus was investigated under in vitro conditions. Relative fresh weight growth (RFWG) of calli, total phenolics (TP) content, endogenous hydrogen peroxide (H₂O₂), malondialdehyde (MDA), proline concentrations, percentage of electrolyte leakage (EL), and some of the antioxidant enzyme activities; such as superoxide dismutase (SOD), and guaiacol peroxidase (POD), catalase (CAT), and ascorbate peroxidase (APX) were measured. Inhibitory effect of high concentrations of BA on antioxidant enzyme activities and RFWG was found. In the presence of BA at 0.1 mg L⁻¹, SOD, POD, and APX activities decreased, while CAT activity increased in comparison with the controls. Exogenous BA treatments higher than 0.1 mg L⁻¹ resulted in an increase in cellular TP, H₂O₂, MDA, proline contents, and percentage of EL, while RFWG of calli decreased. Based on the findings, it may be concluded that only 0.1 mg L⁻¹ BA concentration combined with NAA could play a direct role in reducing the level of free radicals and phenolic production associated with proliferation capacity of grape cells under in vitro conditions. Furthermore, cytokinin was effective in the antioxidative enzyme system, lipid peroxidation, and electrolyte leakage.     

High-frequency vibration improve callus growth via antioxidant enzymes induction in <Emphasis Type="Italic">Hyoscyamus kurdicus</Emphasis>

Effect of high-frequency vibration on growth rate, membrane stability and activities of some antioxidant enzymes were studied in callus tissues of Hyoscyamus kurdicus. Calli initiated from leaf (LE), shoot (SE) and root (RE) explants, and sinusoidal vibrations at 0, 50, 100 and 150 Hz for 30 min were applied on the H. kurdicus calli. Results showed that sinusoidal vibration at 50 and 100 Hz promoted the growth rate as compared to control, and the optimum growth was found at 50 Hz. Sinusoidal vibration increased significantly protein and proline contents and activity of superoxide dismutase (SOD), ascorbate peroxidase (APX) and peroxidase (POX) enzymes, and decreased total carbohydrate, H₂O₂ level and CAT activity as compared to control. Lipid peroxidation also decreased under sinusoidal vibration in all the calli, and the maximum percentage of decrease was observed at 50 Hz. Native polyacrylamide gel electrophoresis indicated different isoform profiles in vibration treated and untreated plants concerning antioxidant enzymes. The responses of different types of calluses were different, and RE callus showed the highest growth, membrane stability and antioxidant enzymes activity as compared to LE and SE calli. These results suggest sinusoidal vibration at optimum frequency could improve callus growth by induction of antioxidative enzymes activity and membrane stability in calli of H. kurdicus.     

Silicon-mediated changes of some physiological and enzymatic parameters symptomatic for oxidative stress in spinach and tomato grown in sodic-B toxic soil

We investigated effect of silicon (Si) on the growth, uptake of sodium (Na), chloride (Cl), boron (B), stomatal resistance (SR), lipid peroxidation (MDA), membrane permeability (MP), lipoxygenase (LOX) activity, proline (PRO) accumulation, H₂O₂ accumulation, non-enzymatic antioxidant activity (AA) and the activities of major antioxidant enzymes (superoxide dismutase, SOD; catalase, CAT and ascorbate peroxidase, APX) of spinach and tomato grown in sodic-B toxic soil. Si applied to the sodic-B toxic soil at 2.5 and 5.0 mM concentrations significantly increased the Si concentration in the plant species and counteracted the deleterious effects of high concentrations of Na, Cl and B on root and shoot growth by lowering the accumulation of these elements in the plants. Stomatal resistance, MP, MDA and the concentrations of H₂O₂ and PRO were higher in the plants grown in sodic-B toxic soil without Si: LOX activity of excised leaves of both species was increased by Si. Antioxidant activities of both species were significantly affected by Si, with the activities of SOD, CAT and APX decreased and AA increased by applied Si. For most of the parameters measured, it was found that 5 mM Si was more effective than the 2.5 mM Si. Based on the present work, it can be concluded that Si alleviates sodicity and B toxicity of the plants grown in sodic-B toxic soil by preventing both oxidative membrane damage and also translocation of Na, Cl and B from root to shoots and/or soil to plant, and lowering the phytotoxic effects of Na, Cl and B within plant tissues. It was concluded that tomato was more responsive to Si than spinach since it was more salt sensitive than spinach. To our knowledge, this is the first report that Si improves the combined salt and B tolerance of spinach and tomato grown in naturally sodic-B toxic soil, and which describes membrane-related parameters and antioxidant responses.     

Recovery of bean plants from boron-induced oxidative damage by zinc supply

The effects of zinc on growth, boron uptake, lipid peroxidation, membrane permeability (MP), lypoxygenase (LOX) activity, proline and H₂O₂ accumulation, and the activities of major antioxidant enzymes (superoxide dismutase (SOD), catalase (CAT), and ascorbate peroxidase (APX)) in bean plants were investigated under greenhouse conditions. Treatments consisted of control, 20 mg/kg B, and 20 mg/kg B plus 20 mg/kg Zn. When the plants were grown with 20 mg/kg Zn, B toxicity was less severe. Zinc supplied to soil counteracted the deleterious effects of B on root and shoot growth. Excess B significantly increased and Zn treatment reduced B concentrations in shoot and root tissues. Applied Zn increased the Zn concentration in the roots and shoots. While the concentrations of H₂O₂ and proline were increased by B toxicity, their concentrations were decreased by Zn supply. Boron toxicity increased the MP, malondialdehyde content, and LOX activity in excised bean leaves. Applied Zn significantly ameliorated the membrane deterioration. Compared with control plants, the activity of SOD was increased while that of CAT was decreased and APX remained unchanged in B-stressed plants. However, application of Zn decreased the SOD and increased the CAT and APX activities under toxic B conditions. It is concluded that Zn supply alleviates B toxicity by preventing oxidative membrane damage. Published in Russian in Fiziologiya Rastenii, 2009, Vol. 56, No. 4, pp. 555–562. This text was submitted by the authors in English.     

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.     

不同生境两种生态型芦苇的抗氧化系统

以分布于甘肃临泽平川乡的两种芦苇生态型——水生芦苇(水芦)和重度盐化草甸芦苇(盐芦)叶片为材料,研究了其抗氧化系统的特征。结果表明,与水芦相比,盐芦中未出现活性氧和MDA(丙二醛)的积累,抗氧化酶SOD(超氧化物歧化酶)、CAT(过氧化氢酶)、POD(过氧化物酶)和APX(抗坏血酸过氧化物酶)的活性显著升高。总抗坏血酸和类胡萝卜素含量在两种生态型芦苇中没有差异,但还原型抗坏血酸和总谷胱甘肽含量在盐芦中显著升高。而且,盐芦的LOX(脂氧合酶)活性比水芦低。这些结果表明,盐芦中有效的抗氧化防御系统对抵抗盐渍胁迫起着重要的作用。此外,盐芦中高活性的Ca^2 -和Mg^2 -ATPase对细胞中过多离子的转运以及避免离子毒害起着重要的作用。     

Interactive effects of salicylic acid and silicon on oxidative damage and antioxidant activity in spinach (<Emphasis Type="Italic">Spinacia oleracea</Emphasis> L. cv. Matador) grown under boron toxicity and salinity

We investigated individual and combined effects of salinity, soil boron (B), silicon (Si) and salicylic acid (SA) on the activities of major antioxidant enzymes (superoxide dismutase, SOD; catalase, CAT and ascorbate peroxidase, APX) and non-enzymatic antioxidants (AA), proline, chlorophyll, anthocyanin, H₂O₂ concentration, stomatal resistance (SR), lipid peroxidation (MDA), membrane permeability (MP), and the uptake of sodium (Na), chloride (Cl), boron and Si of spinach plants. In general, salinity significantly increased H₂O₂ and proline concentrations, antioxidant activity, membrane permeability, lipid peroxidation and SR of the spinach plants, indicating that they were stressed, whereas application of B only increased proline concentration. However, plant fresh weights did not decline with either treatment. The application of Si decreased H₂O₂ and increased the activity of SOD and CAT. The application of SA increased SOD activity. Neither SA nor Si had any effect on the proline concentration, or MP. However, application of Si increased chlorophyll concentration and decreased lipid peroxidation (MDA concentration). Si treatment had no effect on SR. The concentration of B in the tissues, which was strongly increased by B treatment, was decreased by NaCl. As a result of salinity, concentrations of Na⁺ and Cl⁻ ions were increased in the plant tissues, and application of Si slightly increased these concentrations. These results indicate that exogenous Si application increases stress tolerance of spinach, a plant that is naturally reasonably resistant to combined salinity and B toxicity, by the enhancement of antioxidant mechanisms that reduce membrane damage. Exogenous SA has a less obvious effect, although the levels of salinity and boron stress applied were not sufficient in this experiment to reduce plant fresh weight.     

Effects of short-term heat stress on oxidative damage and responses of antioxidant system in Lilium longiflorum

This paper aims to determine the changes in reactive oxygen species (ROS) and the responses of the lily (Lilium longiflorum L.) antioxidant system to short-term high temperatures. Plants were exposed to three levels of heat stress (37°C, 42°C, 47°C) for 10 h when hydrogen peroxide (H₂O₂) and superoxide (O₂⁻) production rate along with membrane injury indexes, and changes in antioxidants were measured. Compared with the control (20°C), electrolyte leakage and MDA concentration varied slightly after 10 h at 37°C and 42°C, while increased significantly at 47°C. During 10 h at 37°C and 42°C, antioxidant enzyme activities, such as SOD, POD, CAT, APX and GR, were stimulated and antioxidants (AsA and GSH concentrations) maintained high levels, which resulted in low levels of O₂⁻ and H₂O₂ concentration. However, after 10 h at 47°C, SOD, APX, GR activities and GSH concentration were similar to the controls, while POD, CAT activities and AsA concentration decreased significantly as compared with the control, concomitant with significant increase in O₂⁻ and H₂O₂ concentrations. In addition, such heat-induced effects on antioxidant enzymes were also confirmed by SOD and POD isoform, as Cu/ZnSOD maintained high stability under heat stress and the intensity of POD isoforms reduced with the duration of heat stress, especially at 47°C. It is concluded that in lily plants, the oxidative damage induced by heat stress was related to the changes in antioxidant enzyme activities and antioxidants.     

Involvement of G6PDH in heat stress tolerance in the calli from Przewalskia tangutica and Nicotiana tabacum

Glucose-6-phosphate dehydrogenase (G6PDH) has been implicated in supplying reduced nicotine amide cofactors for biochemical reactions and in modulating the redox state of cells. In this study, the role of G6PDH in thermotolerance of the calli from Przewalskia tangutica and tobacco (Nicotiana tabacum L.) was investigated. Results showed that Przewalskia tangutica callus was more sensitive to heat stress than tobacco callus. The activity of G6PDH and antioxidant enzymes (ascorbate peroxidase, catalase, peroxidase and superoxide dismutase) in calli from Przewalskia tangutica and tobacco increased after 40 °C treatment, although two calli exhibited a difference in the degree and timing of response to heat stress. When G6PDH was partially inhibited by glucosamine pretreatment, the antioxidant enzyme activities and thermotolerance in both calli significantly decreased. Simultaneously, the heat-induced H₂O₂ content and the plasma membrane NADPH oxidase activity were also reduced. Application of H₂O₂ increased the activity of G6PDH and antioxidant enzymes in both calli. Diphenylene iodonium, a NADPH oxidase inhibitor, counteracted heatinduced H₂O₂ accumulation and reduced the heat-induced activity of G6PDH and antioxidant enzymes. Moreover, exogenous H₂O₂ was effective in restoring the activity of G6PDH and antioxidant enzymes after glucosamine pretreatment. Western blot analysis showed that G6PDH gene expression in both calli was also stimulated by heat and H₂O₂, and blocked by DPI and glucosamine under heat stress. Taken together, under heat stress G6PDH promoted H₂O₂ accumulation via NADPH oxidase and the elevated H₂O₂ was involved in regulating the activity of antioxidant enzymes, which in turn facilitate to maintain the steady-state H₂O₂ level and protect plants from the oxidative damage.     

Opposing roles for superoxide and nitric oxide in the NaCl stress-induced upregulation of antioxidant enzyme activity in cotton callus tissue

The roles of superoxide and NO in the NaCl-induced upregulation on antioxidant enzyme activity were investigated in NaCl-tolerant cotton calli. Both NaCl and paraquat treatments resulted in significant increases in superoxide production. The activities of ascorbate peroxidase (APX), catalase, glutathione reductase (GR), and peroxidase also increased significantly within 2 h after applying the stress. Pre-treatment with the superoxide scavenger, N-acetyl l-cysteine (NAC), completely removed the superoxide and inhibited the upregulation of antioxidant enzyme activity in the tissue treated with either NaCl or paraquat. NaCl stress also resulted in a significant increase in the NO level. Experiments were also carried out to measure antioxidant enzyme activity in cotton calli exposed to NO, the NO producer sodium nitroprusside (SNP), and the NO scavenger 2-phenyl-4,4,5,5-tetramethyl-imidazoline-1-oxyl-3-oxide (PTIO) under different salt stress conditions. The direct addition of NO gas produced no change in the activities of catalase and GR and caused a significant decrease in APX activity when compared to the controls. When the calli was treated with SNP in the absence of NaCl stress, APX and GR activities decreased significantly and catalase activity was only slightly higher than the control. Treatment with SNP in the presence of NaCl stress resulted in a significant decrease in APX activity, and GR and APX activities were not significantly different from those observed in the NaCl treatment alone. In the presence of PTIO, the activities of all three enzymes increased in the presence or absence of NaCl stress. These results suggest that reactive oxygen species (ROS) such as superoxide radicals may serve as signal transduction molecules to switch “on” the early NaCl-induced up-regulation of antioxidant enzyme activity, while NO may play a role in switching “off” the response after other mechanisms in the cascade of events responsible for NaCl tolerance have been activated.