Comparative study on calli from two reed ecotypes under heat stress

The different physiological responses to heat stress in calli from two ecotypes of common reed (Phragmites communis Trin.) plants (dune reed (DR) and swamp reed (SR)) were studied. The relative water content, the relative growth rate, cell viability, membrane permeability (MP), H₂O₂ content, MDA content, proline level, and the activities of enzymes, such as superoxide dismutase (SOD), catalase (CAT), peroxidase (POD), ascorbate peroxidase (APX), glutathione reductase (GR), and lipoxygenase (LOX) were assayed. Results showed that under heat stress, DR callus could maintain the higher relative growth rate and cell viability than SR callus, while H₂O₂ content, MDA content, and MP in SR callus increased more than in DR callus. The activities of antioxidant enzymes, such as SOD, CAT, POD, APX, and GR in two calli were enhanced by high temperature. However, antioxidant enzymes in DR callus showed the higher thermal stability than those in SR callus. LOX activity increased more in SR callus than in DR callus under heat stress. High temperature markedly elevated proline content in DR callus whereas had no effect on that in SR callus. Taken together, DR callus is more thermotolerant than SR callus, which might be due to the higher activity of antioxidant enzymes and proline level compared with SR callus under heat stress.     

cGMP regulates hydrogen peroxide accumulation in calcium-dependent salt resistance pathway in <Emphasis Type="Italic">Arabidopsis thaliana</Emphasis> roots

3′,5′-cyclic guanosine monophosphate (cGMP) is an important second messenger in plants. In the present study, roles of cGMP in salt resistance in Arabidopsis roots were investigated. Arabidopsis roots were sensitive to 100 mM NaCl treatment, displaying a great increase in electrolyte leakage and Na⁺/K⁺ ratio and a decrease in gene expression of the plasma membrane (PM) H⁺-ATPase. However, application of exogenous 8Br-cGMP (an analog of cGMP), H₂O₂ or CaCl₂ alleviated the NaCl-induced injury by maintaining a lower Na⁺/K⁺ ratio and increasing the PM H⁺-ATPase gene expression. In addition, the inhibition of root elongation and seed germination under salt stress was removed by 8Br-cGMP. Further study indicated that 8Br-cGMP-induced higher NADPH levels for PM NADPH oxidase to generate H₂O₂ by regulating glucose-6-phosphate dehydrogenase (G6PDH) activity. The effect of 8Br-cGMP and H₂O₂ on ionic homeostasis was abolished when Ca²⁺ was eliminated by glycol-bis-(2-amino ethyl ether)-N,N,N′,N′-tetraacetic acid (EGTA, a Ca²⁺ chelator) in Arabidopsis roots under salt stress. Taken together, cGMP could regulate H₂O₂ accumulation in salt stress, and Ca²⁺ was necessary in the cGMP-mediated signaling pathway. H₂O₂, as the downstream component of cGMP signaling pathway, stimulated PM H⁺-ATPase gene expression. Thus, ion homeostasis was modulated for salt tolerance.     

In situ Inactivation of the Oxidase Activity of Xylem Peroxidases by H2O2 in the H2O2-Producing Xylem of Zinnia elegans

Zinnia elegans stems with 3,3′, 5, 5′-tetramethylbenzidine (TMB) in the presence and in the absence of catalase reveals the presence of xylem oxidase activities in the H₂O₂-producing lignifying xylem cells. This staining of lignifying xylem cells with TMB is the result of two independent mechanisms: one is the catalase-sensitive (H₂O₂-dependent) peroxidase-mediated oxidation of TMB, and the other the catalase-insensitive (H₂O₂-independent) oxidation of TMB, probably due to the oxidase activity of xylem peroxidases. The response of this TMB-oxidase activity of xylem peroxidases to different exogenous H₂O₂ concentrations was studied, and the results showed that H₂O₂ at high concentrations (100–1,000 mM) clearly acted as an inactivator of this xylem TMB-oxidase activity, although some inhibitory effect could still be appreciated at 10 mM H₂O₂. This xylem TMB-oxidase activity resided in a strongly basic cell wall-bound peroxidase (pl about 10.5). Given such a scenario, it may be concluded that this TMB-oxidase activity of peroxidase is located in tissues capable of sustaining H₂O₂ production, and that the in situ oxidase activity shown by this enzyme is inactivated by high H₂O₂ concentrations. Received 20 April 1999/ Accepted in revised form 16 August 1999     

ROS generation and proline metabolism in calli of halophyte Nitraria tangutorum Bobr. to sodium nitroprusside treatment

Nitric oxide (NO) is a stress factor or a signal molecule involved in various plant physiological and developmental processes. In the present study, the generation of reactive oxygen species and the metabolism of proline due to different sodium nitroprusside (SNP, an NO donor) concentrations were investigated in callus from halophyte Nitraria tangutorum Bobr. Treatment with SNP led to significant increases of hydrogen peroxide (H₂O₂) content and cell viability but notable reductions in hydrogen radical level and lipid peroxidation degree, and superoxide onion (O₂ ^?) content also enhanced in 100 μM SNP-treated calli. Using a chemical inhibitor for plasma membrane (PM) NADPH oxidase diphenylene iodonium (DPI), we found low O₂ ^? generation in untreated and 25 μM SNP-treated calli, whereas in those treated with 100 μM SNP O₂ ^? level exhibited a very little alteration, comparable to the absence of DPI. These suggest a high activity of PM NADPH oxidase in untreated calli. H₂O₂ scavenging enzymes (catalase, peroxidase [POD] and ascorbate peroxidase) and H₂O₂ forming enzymes (superoxide dismutase [SOD], cell wall-POD and diamine oxidase [DAO]) stimulated significantly in calli treated with different SNP concentrations while glutathione reductase activity decreased. In addition, a reduction in proline content was observed in SNP-treated calli. Moreover, different SNP concentrations stimulated proline dehydrogenase (PDH) and ornithine δ-aminotransferase but inhibited r-glutamyl kinase (GK). In conclusion, our results suggest that the increasing H₂O₂ generation was associated with the stimulation of SOD, cell wall-POD and DAO, and that the reduction of proline content might be the consequence of increased PDH activity and decreased GK activity in N. tangutorum Bobr. calli under SNP treatment.     

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.     

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.     

Pre-treatment with H<Subscript>2</Subscript>O<Subscript>2</Subscript> induces salt tolerance in Barley seedlings

Barley seedlings were pre-treated with 1 and 5 μM H₂O₂ for 2 d and then supplied with water or 150 mM NaCl for 4 and 7 d. Exogenous H₂O₂ alone had no effect on the proline, malondialdehyde (MDA) and H₂O₂ contents, decreased catalase (CAT) activity and had no effect on peroxidase (POX) activity. Three new superoxide dismutase (SOD) isoenzymes appeared in the leaves as a result of 1 μM H₂O₂ treatment. NaCl enhanced CAT and POX activity. SOD activity and isoenzyme patterns were changed due to H₂O₂ pre-treatment, NaCl stress and leaf ageing. In pre-treated seedlings the rate of ¹⁴CO₂ fixation was higher and MDA, H₂O₂ and proline contents were lower in comparison to the seedlings subjected directly to NaCl stress. Cl⁻ content in the leaves 4 and 7 d after NaCl supply increased considerably, but less in pre-treated plants. It was suggested that H₂O₂ metabolism is involved as a signal in the processes of barley salt tolerance.     

Exogenous proline alleviates the effects of H2O2-induced oxidative stress in wild almond species

The effect of proline on the antioxidant system in the leaves of eight species of wild almond (Prunus spp.) exposed to H₂O₂-mediated oxidative stress was studied. The levels of endogenous proline (Pro) and hydrogen peroxide, and the activities of total superoxide dismutase (SOD), catalase (CAT), ascorbate peroxidase (APX), glutathione reductase (GR), and guaiacol peroxidase (POD) were measured. The degradation of chlorophyll but not carotenoids occurred in leaves in the solution of 5 mM H₂O₂. An increase in membrane lipid peroxidation was observed in H₂O₂ treatment, as assessed by MDA level and percentage of membrane electrolyte leakage (EL). Significant increases in total SOD and CAT activities, as well as decreases in APX and POD activities, were detected in H₂O₂-treated leaves. The three SOD isoforms showed different behavior, as Mn-SOD activity was enhanced by H₂O₂, whereas Fe-SOD and Cu/Zn-SOD activities were inhibited. In addition, Pro accumulation up to 0.1 ??mol/g fr wt, accompanied by significant decreases in ascorbate and glutathione levels, was observed in H₂O₂-treated leaves. After two different treatments with 10 mM Pro + 5 mM H₂O₂, total SOD and CAT activities were similar to the levels in control plants, while POD and APX activities were higher if compared to the leaves exposed only to H₂O₂. Pro + H₂O₂ treatments also caused a strong reduction in the cellular H₂O₂ and MDA contents and EL. The results showed that Pro could have a key role in protecting against oxidative stress injury of wild almond species by decreasing membrane oxidative damage.     

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.     

Signal regulation of proline metabolism in callus of the halophyte Nitraria tangutorum Bobr. grown under salinity stress

Callus of the halophyte Nitraria tangutorum Bobr. was used to investigate proline metabolism and its signal regulation under salinity stress. Enhanced levels of proline and hydrogen peroxide (H₂O₂) were observed in calli exposed to salinity stress, and elevated levels of calcium (Ca) were detected in early responses to 75?mM NaCl treatment. Additionally, NaCl treatment induced significant elevation of ornithine-??-aminotransferase (OAT) activity, but notable decreases occurred in the activities of glutamyl kinase (GK) and proline dehydrogenase (PDH). H₂O₂ scavenger dimethylthiourea and pyruvate inhibited the accumulation of proline and the stimulation of OAT in salinity-stressed calli. Moreover, the utilization of Ca chelator EGTA and Ca channel blocker verapamil abolished the enhancement of proline level induced by 75?mM NaCl treatment for 3?days. These results suggest that the accumulation of proline is correlated to the increase of OAT activity and the decrease of PDH activity in response to salinity, and that elevated Ca signal during the early stage of NaCl treatment and the excitation of OAT activity resulting from the increase of H₂O₂ generation are essential for proline accumulation in salinity-stressed calli.     

Changes in morphological, photosynthetic and physiological responses of Mono Maple seedlings to enhanced UV-B and to nitrogen addition

In the southeast of the Qinghai-Tibetan Plateau of China, Mono Maple is a common species in reforestation processes. The paper mainly investigated the changes in morphological, photosynthetic and physiological responses of Mono Maple seedlings to UV-B radiation, nitrogen supply and their combination. The experimental design included two levels of UV-B treatments (ambient UV-B, 11.02 KJ m⁻² day⁻¹; enhanced UV-B, 14.33 KJ m⁻² day⁻¹) and two nitrogen levels (0; 20 g N m⁻² a⁻¹)—to determine whether the adverse effects of UV-B on plants are eased by nitrogen supply. Enhanced UV-B caused a marked decline in growth parameters, net photosynthetic rate, and photosynthetic pigments, whereas it induced an increase in reaction oxygen species (hydrogen peroxide accumulation and the rate of superoxide radical production) and malondialdehyde content. Enhance UV-B also induced an increase in antioxidant compounds of Mono Maple, such as UV-B absorbing compounds, proline content, and activities of antioxidant enzymes (peroxidase, superoxide dimutase and catalase). On the other hand, nitrogen supply caused an increase in some growth parameters, net photosynthetic rate, photosynthetic pigments and antioxidant compounds (peroxidase, proline content and UV-B absorbing compounds), and reduced the content of reaction oxygen species (H₂O₂ accumulation, the rate of O₂⁻ production) and malondialdehyde content under ambient UV-B. However, under enhanced UV-B, nitrogen supply inhibited some growth parameters, and increased H₂O₂ accumulation, the rate of O₂⁻ production and MDA content, though proline content, UV-B absorbing compounds and activities of POD and SOD increased. These results implied that enhanced UV-B brought harmful effects on Mono Maple seedlings and nitrogen supply made plants more sensitive to enhanced UV-B, though increased some antioxidant activity.     

Effect of salinity on antioxidant enzymes in calli of the halophyte <Emphasis Type="Italic">Nitraria tangutorum</Emphasis> Bobr.

Nitraria tangutorum Bobr., a typical desert halophyte, plays an important ecological role because of its superior tolerance to severe drought and high salinity. Very little is known about the physiological adaptative mechanism of this species to environmental stresses. The aim of this study was to investigate the changes of antioxidant enzyme activities and the regulatory mechanism of ascorbate peroxidase (APX) activity in the calli from Nitraria tangutorum Bobr. after treatment with different NaCl concentrations. The activities of superoxide dismutase (SOD) and catalase (CAT) significantly increased in the calli treated with NaCl, while the peroxidase activity decreased. APX activity was also elevated significantly in response to NaCl, but the increase was partly abolished by H₂O₂ scavenger dimethylthiourea (DMTU). Furthermore, the excitatory effect of salinity on APX could be alleviated by the addition of exogenous CAT and nicotinamide adenine dinucleotide phosphate (NADPH) oxidase inhibitor diphenylene iodonium, indicating that the modulation of the APX activity in Nitraria tangutorum Bobr. calli might be associated with NADPH oxidase-dependent H₂O₂ generation. Measurement and analysis using fluorescent dye 2′,7′-dichlorodihydrofluorescein diacetate showed the increase of H₂O₂ content in salinity-treated calli. The investigation of NADPH-dependent O₂⁻ production in plasma membrane (PM) vesicles isolated from Nitraria tangutorum Bobr. calli revealed that salinity treatment stimulated NADPH oxidase activity. In conclusion, these results suggest that the higher activities of antioxidant enzymes play an important role in the salt tolerance of Nitraria tangutorum Bobr. calli and that the extracellular production of H₂O₂, depending on the excitation of PM NADPH oxidase, is responsible for enhancing the APX activity in Nitraria tangutorum Bobr. calli under salinity stress.     

Calcium is involved in the abscisic acid-induced ascorbate peroxidase, superoxide dismutase and chilling resistance in Stylosanthes guianensis

The objective of this work was to test whether Ca²⁺, a second messenger in stress response, is involved in ABA-induced antioxidant enzyme activities in Stylosanthes guianensis. Plants were sprayed with abscisic acid (ABA), calcium channel blocker, LaCl₃, calcium chelator, ethylene glycol-bis(β-amino ethyl ether)-N,N,N′, N′-tetraacetid acid (EGTA), and ABA in combination with LaCl₃ or EGTA. Their effects on superoxide dismutase (SOD) and ascorbate peroxidase (APX) activities and chilling resistance were compared. The results showed that ABA decreased electrolyte leakage and lipid peroxidation but increased maximum photochemical efficiency measured as variable to maximum fluorescence ratio (F_v/F_m) under chilling stress. Treatment with LaCl₃ or EGTA alone and in combination with ABA increased electrolyte leakage and lipid peroxidation, decreased Fv/Fm, suggesting that the block in Ca²⁺ signalling decreased chilling resistance of S. guianensis and the ABA-enhanced chilling resistance. ABA-induced SOD and APX activities were suppressed by LaCl₃ or EGTA. The results suggested that Ca²⁺ is involved in the ABA-enhanced chilling resistance and the ABA-induced SOD and APX activities in S. guianensis.     

Changes in photosynthesis and antioxidant defenses of Picea asperata seedlings to enhanced ultraviolet-B and to nitrogen supply

The paper mainly studied the effects of ultraviolet‐B (UV‐B) radiation, nitrogen, and their combination on photosynthesis and antioxidant defenses of Picea asperata seedlings. The experimental design included two levels of UV‐B treatments (ambient UV‐B, 11.02 KJ m⁻² day⁻¹; enhanced UV‐B, 14.33 KJ m⁻² day⁻¹) and two nitrogen levels (0; 20 g m⁻² a⁻¹ N) – to determine whether the adverse effects of UV‐B are eased by supplemental nitrogen. Enhanced UV‐B significantly inhibited plant growth, net photosynthetic rate (A), stomatal conductance to water vapor (Gs), transpiration rate and photosynthetic pigment, and increased intercellular CO₂ concentration, UV‐B absorbing compounds, proline content, malondialdehyde (MDA) content, and activity of antioxidant enzymes (peroxidase (POD), superoxide dimutase, and glutathione reductase). Enhanced UV‐B also reduced needle DW and increased hydrogen peroxide (H₂O₂) content and the rate of superoxide radical (O₂⁻) production only under supplemental nitrogen. On the other hand, supplemental nitrogen increased plant growth, A, Gs, chlorophyll content and activity of antioxidant enzymes (POD, ascorbate peroxidase, and catalase), and reduced MDA content, H₂O₂ content, and the rate of O₂⁻ production only under ambient UV‐B, whereas supplemental nitrogen reduced activity of antioxidant enzymes under enhanced UV‐B. Carotenoids content, proline content, and UV‐B absorbing compounds increased under supplemental nitrogen. Moreover, significant UV‐B × nitrogen interaction was found on plant height, basal diameter, A, chlorophyll a, activity of antioxidant enzymes, H₂O₂, MDA, and proline content. These results implied that supplemental nitrogen was favorable for photosynthesis and antioxidant defenses of P.asperata seedlings under ambient UV‐B. However, supplemental nitrogen made the plants more sensitive to enhanced UV‐B, although some antioxidant indexes increased.     

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.     

Hexanoic acid 2-(diethylamino)ethyl ester enhances chilling tolerance in strawberry seedlings by impact on photosynthesis and antioxidants

Strawberry (Fragaria ananassa Duch.) seedlings were pretreated with hexanoic acid 2-(diethylamino)ethyl ester (DA-6) in concentrations of 0, 10, 20 and 40 mg dm⁻³ and then subjected to chilling and rewarming. The effects of applied DA-6 on the generation of reactive oxygen species (O₂ ⁻, H₂O₂), lipid peroxidation, proline accumulation and photosynthesis were evaluated. Pretreatment with DA-6 alleviated the inhibition of superoxide dismutase (SOD), catalase (CAT) and ascorbate peroxidase (APX) activities caused by chilling stress thus reducing O₂ ⁻ and H₂O₂ production and lipid peroxidation in pretreated plants. DA-6 pretreatment also accelerated accumulation of proline and reduce the decrease in proline content after rewarming. DA-6 pretreatment increases maximum quantum yield of photosystem 2 (F_v/F_m), actual photochemical efficiency of photosystem 2 (Φ_PS2), photochemical quenching coefficient (qP) and net photosynthetic rate (P_N) and decreases non-photochemical quenching coefficient (qNP) of the seedlings under chilling stress. DA-6 pretreatment also increased the recovery rate of photosynthesis after rewarming.     

Arsenic-induced root growth inhibition in mung bean (<Emphasis Type="Italic">Phaseolus aureus</Emphasis> Roxb.) is due to oxidative stress resulting from enhanced lipid peroxidation

Arsenic (As) toxicity and its biochemical effects have been mostly evaluated in ferns and a few higher plants. In this study, we investigated the effect of As (10.0 and 50.0 μM) on seedling growth, root anatomy, lipid peroxidation (malondialdehyde and conjugated dienes), electrolyte leakage, H₂O₂ content, root oxidizability and the activities of antioxidant enzymes in mung bean (Phaseolus aureus Roxb.). Arsenic significantly enhanced lipid peroxidation (by 52% at 50.0 μM As), electrolyte leakage and oxidizability in roots. However, there was no significant change in H₂O₂ content. Arsenic toxicity was associated with an increase in the activities of superoxide dismutase (SOD), guaiacol peroxidase (GPX) and glutathione reductase (GR). In response to 50.0 μM As, the activities of SOD and GR increased by over 60% and 90%, respectively. At 10.0 μM As, the activity of ascorbate peroxidase (APX) increased by 83%, whereas at 50.0 μM it declined significantly. The catalase (CAT) activity, on the other hand, decreased in response to As exposure, and it corresponded to the observed decrease in H₂O₂ content. We conclude that As causes a reduction in root elongation by inducing an oxidative stress that is related to enhanced lipid peroxidation, but not to H₂O₂ accumulation.     

Effect of salt stress on some physio-biochemical traits and antioxidative enzymes of two Brassica species under callus culture

The changes in lipid peroxidation, H₂O₂, proline, protein, involvement of different antioxidant systems (catalase, guaiacol peroxidase, ascorbate peroxidase) and callus-related traits were investigated under salt stress in the callus of two different ploidy levels of Brassica including B. juncea and B. oleracea. The calluses of B. juncea genotypes were less sensitive to NaCl stress in comparison with those of B. oleracea while increasing the concentrations of NaCl from 0 to 200 mM. Tetraploid genotype (B. juncea cr3356) showed a significant increase in the contents of protein and proline, and guaiacol peroxidase activity and catalase enzymes at higher salinity levels. In addition, a significant decrease occurred in the amount of H₂O₂ and malondialdehyde along with increasing the salinity intensity. Diploid cultivar (B. oleracea bra 2828) had the lowest enzymatic activities and the highest content of H₂O₂ and malondialdehyde along with an increase in the salinity level. Therefore, this genotype was identified as the most sensitive cultivar under the salinity stress. The salinity resistance difference between diploid and amphidiploid species could be attributed to the differences in the ploidy level of these species. This result underlines the fact that the tetraploid genome of B. oleracea could be considered as a suitable candidate for production under salinity conditions through maintaining higher activities of antioxidant enzymes.

     

外源H2O2处理对唐古特白刺愈伤组织脯氨酸代谢的影响

以唐古特白刺（Nitraria tangutorum Bobr.）愈伤组织为材料,研究外源H₂O₂（2和10 μmol·L^-1）处理下其脯氨酸含量及相关代谢酶活性的变化,试图从细胞水平揭示H₂O₂影响脯氨酸代谢的生理机制。结果显示,2和10 μmol·L^-1 H₂O₂处理24 h使唐古特白刺愈伤组织脯氨酸含量分别变为对照的112%和92%,而处理72 h后,脯氨酸含量增加为对照的141%和119%;与对照相比,外源H₂O₂处理诱导愈伤组织脯氨酸脱氢酶活性降低,而谷氨酸激酶活性升高,但鸟氨酸转氨酶活性无显著变化;此外,H₂O₂处理使唐古特白刺愈伤组织内源性H₂O₂含量升高。结果表明,外源H₂O₂诱导了唐古特白刺愈伤组织H₂O₂含量的增高和脯氨酸的积累,且H₂O₂处理下脯氨酸脱氢酶活性的降低及谷氨酸激酶的升高与愈伤组织脯氨酸的积累有关。     

Chilling,oxidative stress and antioxidant responses inArabidopsis thaliana callus

Chilling ofArabidopsis thaliana (L.) Heynh. callus tissue to 4 °C led to conditions of oxidative stress, as indicated by increased levels of the products of peroxidative damage to cell membranes. Cellular H₂O₂ was also observed to increase initially upon chilling but by day 8 cellular levels had declined to below control levels. Although levels of catalase activity remained similar to those in unchilled tissue, activity of ascorbate peroxidase increased between days 4 and 8 of chilling to 4 °C. In callus held at 23 °C, levels of reduced glutathione remained static whereas they rose in callus held at 4 °C. Levels of oxidised glutathione were initially low but increased significantly by day 4 in the chilled callus. At 23 °C, however, levels of oxidised glutathione remained low. Between days 1 and 3 at 4 °C, levels of glutathione reductase activity increased but by day 8 glutathione reductase activity was similar to that in cells held at 23 °C. Exposure of callus to abscisic acid at 23 °C also led to increased activities of ascorbate peroxidase and glutathione reductase.Abbreviations ABA abscisic acid - GSH reduced glutathione - GSSG oxidised glutathione - TTC 2,35-triphenyltetrazolium chloride This work is supported by a grant from the Biotechnology and Biological Sciences Research Council.