Differential biochemical responses of wheat shoots and roots to nickel stress: antioxidative reactions and proline accumulation

Wheat (Triticum aestivum L. cv. ‘Zyta’) seedlings were treated with 10, 100 and 200 μM Ni. Tissue Ni accumulation, length, relative water content (RWC), proline and H₂O₂ concentrations as well as the activities of superoxide dismutase (SOD), catalase (CAT), ascorbate peroxidase (APX), guaiacol peroxidase (POD) and glutathione S-transferase (GST) were studied in the shoots and roots after 6 days of Ni exposure. Treatment with Ni, except for its lowest concentration, resulted in a significant reduction in wheat growth. In comparison to the shoots, the roots showed greater inhibition of elongation, which corresponded with higher accumulation of Ni in these organs. Both shoots and roots responded to Ni application with a decrease in RWC and enhancement in proline concentration. Greater dehydration of the shoot tissue was accompanied by more intense accumulation of proline. Treatment of the wheat seedlings with the highest concentration of Ni led to about 60% increase in H₂O₂ concentration in both studied organs. Apart from CAT, constitutive activities of antioxidative enzymes were much higher in the roots than in the shoots. Exposure of the seedlings to Ni resulted in SOD activity decline, which was more marked in the roots. While the shoots showed a substantial decrease (up to 30%) in CAT activity, in the roots the activity of this enzyme remained unchanged. After Ni application APX, POD and GST activities increased several-fold in the shoots, whereas in the roots they were not significantly altered. The results suggest that differential antioxidative responses of the shoots and roots of wheat seedlings to Ni stress might be related to diverse constitutive levels of antioxidant enzyme activities in both organs.     

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.     

Nickel-induced oxidative stress and the role of antioxidant defence in rice seedlings

Seedlings of rice (Oryza sativa L.) cv. Pant-12 grown in sand cultures containing 200 and 400 μM NiSO₄, showed a decrease in length and fresh weight of roots and shoots. Nickel was readily taken up by rice seedlings and the concentration was higher in roots than shoots. Nickel-treated seedlings showed increased rates of superoxide anion (O₂ ^•− ) production, elevated levels of H₂O₂ and thiobarbituric acid reactive substances (TBARS) demonstrating enhanced lipid peroxidation, and a decline in protein thiol levels indicative of increased protein oxidation compared to controls. With progressively higher Ni concentrations, non-protein thiol and ascorbate (AsA) increased, whereas the level of low-molecular-weight thiols (such as glutathione and hydroxyl-methyl glutathione), the ratio of these thiols to their corresponding disulphides, and the ratio of AsA to dehydroascorbic acid declined in the seedlings. Among the antioxidant enzymes studied, the activities of all isoforms of superoxide dismutase (Cu-Zn SOD, Mn SOD and Fe SOD), guaiacol peroxidases (GPX) and ascorbate peroxidase (APX) increased in Ni-treated seedlings, while no clear alteration in catalase activity was evident. Activity of the ascorbate-glutathione cycle enzymes monodehydroascorbate reductase (MDHAR), dehydroascorbate reductase (DHAR) and glutathione reductase (GR)—significantly increased in Ni-treated seedlings. However such increase was apparently insufficient to maintain the intracellular redox balance. Results suggest that Ni induces oxidative stress in rice plants, resulting in enhanced lipid peroxidation and decline in protein thiol levels, and that (hydroxyl-methyl) glutathione and AsA in conjunction with Cu-Zn SOD, GPX and APX are involved in stress response.     

Effect of nickel on ROS content and antioxidative enzyme activities in wheat leaves

Influence of 100 μM Ni on growth, Ni accumulation,, H₂O₂ and lipid peroxides contents as well as the activities of superoxide dismutase (SOD), catalase (CAT), ascorbate peroxidase (APX), guaiacol peroxidase (POD) and glutathione peroxidase (GSH-Px) were studied in the leaves of wheat plants on the 3rd, 6th and 9th days after treatment. Exposure of the plants to Ni for only 3 days led to almost 200-fold increase in this metal concentration in the leaf tissue but later the rate of Ni accumulation was much slower. Length and fresh weight of the leaves were substantially reduced, up to 25% and 39%, respectively at the end of experiment. Visible symptoms of Ni toxicity: chlorosis and necrosis were observed following the 3rd day. Treatment with Ni resulted in the increase in and H₂O₂ contents in the leaves. Both showed their highest values, approximately 250% of those of the control, on the 3rd day and then their levels decreased but still markedly exceeded the control values. SOD and CAT activities decreased significantly in response to Ni treatment, however a several-fold increase in APX and POD activities was found. No significant changes in lipid peroxides content were observed in the leaves after Ni application. The activity of GSH-Px showed a 29% induction on the 3rd day. Our results indicated that despite prolonged increases in and H₂O₂ levels, oxidative damage, measured as the level of lipid peroxidation, did not occur in the leaves of Ni-treated wheat.     

IBA-induced changes in antioxidant enzymes during adventitious rooting in mung bean seedlings: The role of H2O2

The changes in antioxidant enzyme activity during the induction of adventitious roots in mung bean seedlings treated with Indole-3-butyric acid (IBA), hydrogen peroxide (H₂O₂), ascorbic acid (ASA) and diphenylene iodonium (DPI) were investigated. As compared with the controls, treatments of seedlings with 10 μM IBA significantly decreased POD activity by 55% and 49.6% at 3 h and 12 h of incubation, respectively, and significantly increased by 49.8% at 36 h of incubation; treatments of seedlings with 10 mM H₂O₂ significantly decreased POD activity by 42%, 60%, 39% and 38% at 3 h, 12 h, 24 h and 48 h of incubation, respectively, the changes in POD activity were coincident with those in IBA-treated seedlings during the 0–12 h incubation period; treatments of seedlings with 2 mM ASA significantly decreased APX activities by 27% only at 3 h of incubation, the varying trend of POD activity was similar to incubation with water; 10 μM DPI treatments significantly decreased POD activity by 42%, 40%, 54% and 28% at 3 h, 6 h, 12 h and 48 h of treatment, respectively. CAT activities remained at relatively stable levels and no major changes occurred from 0 h to 48 h during the incubation phase of adventitious rooting. The results may imply that CAT, an H₂O₂-metabolizing enzyme, is inactivated by H₂O₂ during the formation of adventitious roots. As compared with the controls, IBA treatments significantly decreased APX activities by 48%, 53% and 66% at 3 h, 9 h and 12 h of treatment, respectively; H₂O₂ treatments significantly decreased APX activities by 59%, 51% and 57% at 3 h, 12 h and 36 h of incubation, respectively; ASA treatments significantly decreased APX activities by 37% only at 3 h of incubation; DPI treatments significantly decreased APX activities by 54%, 53% and 63% at 3 h, 6 h and 12 h of incubation, respectively, and significantly increased APX activity by 106% at 24 h. These results indicated that the influence of IBA, H₂O₂, ASA and DPI on the changes in APX activity were the same as on the changes in POD activity. Furthermore, similar trends in the changes of APX activity and POD activity were observed during the induction and initiation rooting phase. This finding implies that APX and POD serve the same functions, possibly related to the level of H₂O₂, during the formation of adventitious roots. The early decrease of POD and APX activities in the initiation phase of IBA- and H₂O₂-treated seedlings may be one mechanism underlying the IBA- and H₂O₂-mediated facilitation of adventitious rooting.     

Antioxidant system activation by mercury in Pfaffia glomerata plantlets

Oxidative stress caused by mercury (Hg) was investigated in Pfaffia glomerata plantlets grown in nutrient solution using sand as substrate. Thirty-day-old acclimated plants were treated for 9 days with four Hg levels (0, 1, 25 and 50 μM) in the substrate. Parameters such as growth, tissue Hg concentration, toxicity indicators (δ-aminolevulinic acid dehidratase, δ-ALA-D, activity), oxidative damage markers (TBARS, lipid peroxidation, and H₂O₂ concentration) and enzymatic (superoxide dismutase, SOD, catalase, CAT, and ascorbate peroxidase, APX) and non-enzymatic (non-protein thiols, NPSH, ascorbic acid, AsA, and proline concentration) antioxidants were investigated. Tissue Hg concentration increased with Hg levels. Root and shoot fresh weight and δ-ALA-D activity were significantly decreased at 50 μM Hg, and chlorophyll and carotenoid concentration were not affected. Shoot H₂O₂ concentration increased curvilinearly with Hg levels, whereas lipid peroxidation increased at 25 and 50 μM Hg, respectively, in roots and shoots. SOD activity showed a straight correlation with H₂O₂ concentration, whereas CAT activity increased only in shoots at 1 and 50 μM Hg. Shoot APX activity was either decreased at 1 μM Hg or increased at 50 μM Hg. Conversely, root APX activity was only increased at 1 μM Hg. In general, AsA, NPSH and proline concentrations increased upon addition of Hg, with the exception of proline in roots, which decreased. These changes in enzymatic and non-enzymatic antioxidants had a significant protective effect on P. glomerata plantlets under mild Hg-stressed conditions.     

Antioxidant response of wheat roots to drought acclimation

Wheat (Triticum aestivum L.) seedlings of a drought-resistant cv. C306 were subjected to severe water deficit directly or through stress cycles of increasing intensity with intermittent recovery periods. The antioxidant defense in terms of redox metabolites and enzymes in root cells and mitochondria was examined in relation to membrane damage. Acclimated seedlings exhibited higher relative water content and were able to limit the accumulation of H₂O₂ and membrane damage during subsequent severe water stress conditions. This was due to systematic up-regulation of superoxide dismutase, ascorbate peroxidase (APX), catalase, peroxidases, and ascorbate–glutathione cycle components at both the whole cell level as well as in mitochondria. In contrast, direct exposure of severe water stress to non-acclimated seedlings caused greater water loss, excessive accumulation of H₂O₂ followed by elevated lipid peroxidation due to the poor antioxidant enzyme response particularly of APX, monodehydroascorbate reductase, dehydroascorbate reductase, glutathione reductase, and ascorbate–glutathione redox balance. Mitochondrial antioxidant defense was found to be better than the cellular defense in non-acclimated roots. Termination of stress followed by rewatering leads to a rapid enhancement in all the antioxidant defense components in non-acclimated roots, which suggested that the excess levels of H₂O₂ during severe water stress conditions might have inhibited or down-regulated the antioxidant enzymes. Hence, drought acclimation conferred enhanced tolerance toward oxidative stress in the root tissue of wheat seedlings due to both reactive oxygen species restriction and well-coordinated induction of antioxidant defense.     

Medium salt strength induced changes in growth, physiology and secondary metabolite content in adventitious roots of Morinda citrifolia: the role of antioxidant enzymes and phenylalanine ammonia lyase

In an attempt to improve growth and secondary metabolite production, and to understand the possible mechanism involved in relation to the changes in physiology and activities of antioxidant enzymes, we cultured Morinda citrifolia adventitious roots in different strength (0.25, 0.50, 0.75, 1.0, 1.5 and 2.0) of Murashige and Skoog (MS) medium supplemented with 5 mg l⁻¹ indole butyric acid and 30 g l⁻¹ sucrose. Quarter-strength MS medium was proven suitable for the production of both root biomass and secondary metabolites [anthraquinone (AQ), phenolics and flavonoids]. With the increasing salt strength, root growth and AQ accumulation decreased significantly. Higher (1.5 and 2 MS) salt strength provoked osmotic stress resulted in more than twofold free proline accumulation than lower salt strength treated roots and induced free radical scavenging activity. Phenylalanine ammonia lyase activity showed a positive correlation in relation to salt strength that leads to an increase in phenol biosynthesis in expense of AQ formation. The elevated catalase (CAT), guaiacol peroxidase (G-POD) and superoxide dismutase activities and decreased ascorbate peroxidase (APX) activities were observed in roots treated with 2.0 MS. On the other hand, APX activity was strongly activated along with considerable increase in CAT activity at 0.25 MS treated culture. However, the joint functions of CAT, G-POD and APX at 0.25 MS treated cultures were efficient to eliminate the potential danger of hydrogen peroxide (H₂O₂) as evidenced from low H₂O₂ accumulation and low level of lipid peroxidation.     

Calcium protects <Emphasis Type="Italic">Trifolium repens</Emphasis> L. seedlings against cadmium stress

The effect of calcium (Ca²⁺) on Trifolium repens L. seedlings subjected to cadmium (Cd²⁺) stress was studied by investigating plant growth and changes in activity of antioxidative enzymes. Physiological analysis was carried out on seedlings cultured for 2 weeks on half-strength Hoagland medium with Cd²⁺ concentrations of 0, 400 and 600 μM, and on corresponding medium supplied with CaCl₂ (5 mM). Exposure to increasing Cd²⁺ reduced the fresh weight of the upper part (stems + leaves) of the seedlings more strongly than that of the root system. In both parts of T. repens seedlings H₂O₂ level and lipid peroxidation increased. In the upper part, Cd²⁺ exposure led to a significant decrease in the activity of superoxide dismutase, catalase and glutathione peroxidase and an increase in ascorbate peroxidase activity. In contrast, the roots showed an increase in the activity of antioxidative enzymes under Cd²⁺ stress. Ca²⁺ addition to medium reduced the Cd²⁺ accumulation, and considerably reversed the Cd²⁺-induced decrease in fresh mass as well as the changes in lipid peroxidation in the both parts of T. repens seedlings. Ca²⁺ application diminished the Cd²⁺ effect on the activity of antioxidative enzymes in the upper part, even though it did not significantly affect these enzymes in the roots. So the possible mechanisms for the action of Ca²⁺ in Cd²⁺ stress were considered to reduce Cd²⁺ accumulation, alleviate lipid peroxidation and promote activity of antioxidative enzymes.     

Antioxidant responses of shoots and roots of lentil to NaCl-salinity stress

The effect of salt stress (100 mM and 200 mM NaCl) on antioxidant responses in shoots and roots of 14-day-old lentil (Lens culinaris M.) seedlings was investigated. Salt stress caused a significant decrease in length, wet-dry weight and an increase in proline content of both shoot and root tissues. In leaf tissues, high salinity treatment resulted in a 4.4 fold increase in H₂O₂ content which was accompanied by a significant level of lipid peroxidation and an increase in electrolyte leakage. Root tissues were less affected with respect to these parameters. Leaf tissue extracts exhibited four activity bands, of which two were identified as Cu/Zn-SOD and others as Fe-SOD and Mn-SOD. Fe-SOD activity was missing in root extracts. In both tissues Cu/Zn-SOD activity comprised 70–75% of total SOD activity. Salt stress did not cause a significant increase in total SOD activity of leaf tissues but a significant enhancement (88%) was observed in roots mainly due to an enhancement in Cu/ZnSOD isoforms. Compared to leaf tissues a significantly higher constitutive ascorbate peroxidase (APX) and glutathion reductase (GR) activity was observed in root tissues. Upon salt stress no significant change in the activity of APX, catalase (CAT) and GR was observed in root tissues but a higher APX activity was present when compared to leaf tissues. On the other hand, in leaf tissues, with the exception of CAT, salt stress caused significant enhancement in the activity of other antioxidant enzymes. These results suggested that, root tissues of lentil are protected better from NaCl stress induced oxidative damage due to enhanced total SOD activity together with a higher level of APX activity under salinity stress. To our knowledge this is the first report describing antioxidant enzyme activities in lentil.     

Differences in Cowpea Root Growth Triggered by Salinity and Dehydration are Associated with Oxidative Modulation Involving Types I and III Peroxidases and Apoplastic Ascorbate

The aim of this work was to investigate the balance between the activities of ascorbate peroxidase (APX) and phenol peroxidases (POD) and cowpea root growth in response to dehydration and salt stress. Root growth and indicators of oxidative response were markedly changed in response to salinity and dehydration. Salt treatment strongly inhibited root elongation, which was associated with an increase in lignin content and a significant decrease in the concentrations of apoplastic hydrogen peroxide (H₂O₂) and ascorbate. In conditions of extreme salinity, cytosol–APX activity was significantly decreased. In contrast, cell-wall POD activity was greatly increased, whereas lipid peroxidation was unchanged. These results indicate that POD could be involved in both H₂O₂ scavenging and the inhibition of root elongation under high salinity. In contrast, dehydration stimulated primary root elongation and increased lipid peroxidation and apoplastic ascorbate content, but it did not change APX and POD activities or H₂O₂ concentration. When cowpea roots were subjected to salinity followed by dehydration, the water and pressure potentials were decreased, and lipid peroxidation was markedly increased, highlighting the additive nature of the inhibitory effects caused by salt and dehydration. The proline concentration was markedly increased by dehydration alone, as well as by salt followed by dehydration, suggesting a possible role for proline in osmotic adjustment. Salinity and dehydration induce contrasting responses in the growth and morphology of cowpea roots. These effects are associated with different types of oxidative modulation involving cytosolic-APX and cell-wall POD activities and apoplast H₂O₂ and ascorbate levels.     

Effects of aluminum on superoxide dismutase and peroxidase activities,and lipid peroxidation in the roots and calluses of soybeans differing in aluminum tolerance

The seedlings of two soybean genotypes, Al-tolerant PI 416937 (PI) and Al-sensitive Young, were cultured in the solution containing 0, 25 or 50 μM Al (AlCl₃·6H₂O) for 24, 36 or 48 h in the hydroponics, and the calluses induced from two genotypes were cultured in medium containing 0, 10, 50 or 100 μM Al for 5, 10 or 15 days, respectively. The effects of Al on growth of seedling roots and calluses, antioxidant enzyme activities of superoxide dismutase (SOD) and peroxidase (POD) and lipid peroxidation were investigated. Under Al stress, PI was more tolerant to Al toxicity than Young at both intact plant and tissue levels and lower concentrations of Al significantly stimulated the root and callus growth of PI. Al application enhanced the activities of SOD and POD and lipid peroxidation in both roots and calluses of two genotypes. Although the differences of SOD activities between two genotypes in response to Al toxicity depended on Al concentration and durations of treatment, SOD activities in the roots of PI were higher than those in the roots of corresponding Young in the presence of Al for 36 or 48 h. Meanwhile, the POD activities in PI roots increased as the Al levels and durations of treatment increased, significantly higher than those in the corresponding Young roots. Moreover, Al-treated PI had significantly lower lipid peroxidation than Young at both root and callus levels. These results suggest that the enhanced antioxidant-related enzyme activities and reduced lipid peroxidation in PI might be one of Al-tolerant mechanisms.     

山黧豆根系对H2O2诱导氧化胁迫的生理应答

以水培7d苗龄的山黧豆幼苗为材料,向水培溶液中施加不同浓度H2O2处理山黧豆幼苗24h,分析山黧豆根系受氧化胁迫的程度与抗氧化系统的应答特征,以揭示山黧豆对氧化胁迫的耐受机制。结果显示:(1)随外源H2O2处理浓度的不断增加,山黧豆幼苗侧根的数目无显著变化,而其根的鲜重则显著降低。(2)同时,根系组织的内源H2O2染色范围和程度显著增高,但根尖区域始终保持较低水平的H2O2;相反,O-·2染色范围和程度明显减少,根尖区域却始终保持较高水平的O-·2。(3)同期根系抗坏血酸(ASC)含量及过氧化氢酶(CAT)、过氧化物酶(POD)与抗坏血酸过氧化物酶(APX)的活性均表现出了先升高后降低的趋势,而超氧化物歧化酶(SOD)一直表现为持续上升的趋势。研究表明,在外源H2O2胁迫条件下,山黧豆根系O-·2的积累可能与其生长和活力呈正相关,而根系H2O2的积累则与其受氧化胁迫程度呈正相关;低浓度的H2O2处理可以提高山黧豆抗氧化系统对体内活性氧的清除能力。     

IBA-induced changes in antioxidant enzymes during adventitious rooting in mung bean seedlings: The role of H2O2

The changes in antioxidant enzyme activity during the induction of adventitious roots in mung bean seedlings treated with Indole-3-butyric acid (IBA), hydrogen peroxide (H₂O₂), ascorbic acid (ASA) and diphenylene iodonium (DPI) were investigated. As compared with the controls, treatments of seedlings with 10 μM IBA significantly decreased POD activity by 55% and 49.6% at 3 h and 12 h of incubation, respectively, and significantly increased by 49.8% at 36 h of incubation; treatments of seedlings with 10 mM H₂O₂ significantly decreased POD activity by 42%, 60%, 39% and 38% at 3 h, 12 h, 24 h and 48 h of incubation, respectively, the changes in POD activity were coincident with those in IBA-treated seedlings during the 0–12 h incubation period; treatments of seedlings with 2 mM ASA significantly decreased APX activities by 27% only at 3 h of incubation, the varying trend of POD activity was similar to incubation with water; 10 μM DPI treatments significantly decreased POD activity by 42%, 40%, 54% and 28% at 3 h, 6 h, 12 h and 48 h of treatment, respectively. CAT activities remained at relatively stable levels and no major changes occurred from 0 h to 48 h during the incubation phase of adventitious rooting. The results may imply that CAT, an H₂O₂-metabolizing enzyme, is inactivated by H₂O₂ during the formation of adventitious roots. As compared with the controls, IBA treatments significantly decreased APX activities by 48%, 53% and 66% at 3 h, 9 h and 12 h of treatment, respectively; H₂O₂ treatments significantly decreased APX activities by 59%, 51% and 57% at 3 h, 12 h and 36 h of incubation, respectively; ASA treatments significantly decreased APX activities by 37% only at 3 h of incubation; DPI treatments significantly decreased APX activities by 54%, 53% and 63% at 3 h, 6 h and 12 h of incubation, respectively, and significantly increased APX activity by 106% at 24 h. These results indicated that the influence of IBA, H₂O₂, ASA and DPI on the changes in APX activity were the same as on the changes in POD activity. Furthermore, similar trends in the changes of APX activity and POD activity were observed during the induction and initiation rooting phase. This finding implies that APX and POD serve the same functions, possibly related to the level of H₂O₂, during the formation of adventitious roots. The early decrease of POD and APX activities in the initiation phase of IBA- and H₂O₂-treated seedlings may be one mechanism underlying the IBA- and H₂O₂-mediated facilitation of adventitious rooting.     

Salinity induced the changes of root growth and antioxidative responses in two wheat cultivars

This study aimed to investigate the inhibitory mechanism of root growth and to compare antioxidative responses in two wheat cultivars, drought-tolerant Ningchun and drought-sensitive Xihan, exposed to different NaCl concentrations. Ningchun exhibited lower germination rate, seedling growth, and lipid peroxidation than Xihan when exposed to salinity. The loss of cell viability was correlated with the inhibition of root growth induced by NaCl stress. Moreover, treatments with H₂O₂ scavenger dimethylthiourea and catalase (CAT) partly blocked salinity-induced negative effects on root growth and cell viability. Besides, the enhancement of superoxide radical and H₂O₂ levels, and the stimulation of CAT and diamine oxidase (DAO) as well as the inhibition of glutathione reductase (GR) were observed in two wheat roots treated with salinity. However, hydroxyl radical content increased only in Xihan roots under NaCl treatment, and the changes of soluble peroxidase (POD), ascorbate peroxidase (APX), superoxide dismutase (SOD), and cell-wall-bound POD activities were different in drought-tolerant Ningchun and drought-sensitive Xihan exposed to different NaCl concentrations. In conclusion, salinity might induce the loss of cell viability via a pathway associated with extracellular H₂O₂ generation, which was the primary reason leading to the inhibition of root growth in two wheat cultivars. Here, it was also suggested that increased H₂O₂ accumulation in the roots of drought-tolerant Ningchun might be due to decreased POD and GR activities as well as enhanced cell-wall-bound POD and DAO ones, while the inhibition of APX and GR as well as the stimulation of SOD and DAO was responsible for the elevation of H₂O₂ level in drought-sensitive Xihan roots.     

Heat shock-mediated H2O2 accumulation and protection against Cd toxicity in rice seedlings

Rice (Oryza sativa L.) seedlings stressed with CdCl₂ (0.5 mM or 50 μM) showed typical Cd toxicity (leaf chlorosis, decrease in chlorophyll content, or increase in H₂O₂ and malondialdehyde contents). Rice seedlings pretreated with heat shock at 45°C (HS) for 2 or 3 h were protected against subsequent Cd stress. Rice seedlings pretreated with HS had similar Cd concentration in leaves caused by CdCl₂ as those non-HS. The content of H₂O₂ increased in leaves 1 h after HS exposure. However, APX and GR activities were higher in HS-treated leaves than their respective control, and it occurred after 2 h of HS treatment. Pretreatment of rice seedlings with H₂O₂ under non-HS conditions resulted in an increase in APX, GR, and CAT activities and protected rice seedlings from subsequent Cd stress. HS-induced H₂O₂ production and protection against subsequent Cd stress can be counteracted by imidazole, an inhibitor of NADPH oxidase complex. Results of the present study suggest that early accumulation of H₂O₂ during HS signals the increase in APX and GR activities, which in turn prevents rice seedlings from Cd-caused oxidative damage.     

Lack of K-Dependent Oxidative Stress in Cotton Roots Following Coronatine-Induced ROS Accumulation

Coronatine [COR] is a novel type of plant growth regulator with similarities in structure and property to jasmonate. The objective of this study was to examine the relationship between increased root vitality induced by 10nM COR and reactive oxygen species scavenging under potassium (K)-replete (2.5mM) and K-deficient (0.05mM) conditions in hydroponic cultured cotton seedlings. K-replete and K-deficient conditions increased root vitality by 2.7- and 3.5-fold, respectively. COR treatment significantly decreased lipid peroxidation in cotton seedlings determined by reduction in MDA levels. These results suggest that COR improves the functioning of both enzymatic and non-enzymatic antioxidant systems. Under K-replete and K-deficient conditions, COR significantly increased the activities of antioxidant enzymes SOD (only for K-repletion), CAT, GPX, and APX comparing; COR also significantly increased DPPH-radical scavenging activity. However, COR led to 1.6- and 1.7-fold increases in superoxide anion (O₂^•-) concentrations, and 5.7- and 2.1-fold increases in hydrogen peroxide (H₂O₂) levels, respectively. Additionally, COR intensified the DAB staining of H₂O₂ and the NBT staining of O₂^•-. Therefore, our results reveal that COR-induced ROS accumulation stimulates the activities of most antioxidant enzymes but does not induce oxidative stress in cotton roots.     

Cadmium toxicity and translocation in rice seedlings are reduced by hydrogen peroxide pretreatment

A hydroponic experiment was carried out to study the role of hydrogen peroxide (H₂O₂) in enhancing tolerance and reducing translocation of cadmium (Cd) in rice seedlings. Plant growth (length and biomass of shoot and root) was significantly repressed by Cd exposure. However, pretreatment with 100 μM H₂O₂ for 1d mitigated Cd stress by inducing enzyme activities for antioxidation (e.g., superoxide dismutase (SOD), catalase (CAT), guaiacol peroxidase (GPX), ascorbate peroxidase (APX)) and detoxification (e.g., glutathione S-transferase (GST)) as well as by elevating contents of reduced glutathione (GSH) and ascorbic acid (AsA). As a result, H₂O₂ and malondialdehyde (MDA) content decreased in plants and the seedling growth was less inhibited. On the other hand, H₂O₂ pretreatment decreased Cd concentration in shoots, thus lowered the ratio of Cd concentration in shoots and roots (S/R), indicating that H₂O₂ may affect Cd distribution in rice seedlings. The improved Cd tolerance is partly due to an enhanced antioxidative system that efficiently prevents the accumulation of H₂O₂ during Cd stress. Increased Cd sequestration in rice roots may contribute to the decline of Cd translocation.     

Dimethylthiourea, a hydrogen peroxide trap, partially prevents stress effects and ascorbate peroxidase increase in spermidine-treated maize roots

Inhibition of root growth and accumulation of putrescine caused by exogenous spermidine in roots of maize seedlings (Zea mays L., cv Samodek) were partially prevented by a concomitant treatment with dimethylthiourea (DMTU), that traps H₂O₂ produced from spermidine by the activity of polyamine oxidase (PAO) in the apoplast. Treatment with spermidine caused a strong increase of ascorbate peroxidase (APX) gene expression, that was induced to a lesser extent by removing spermidine-generated H₂O₂ by DMTU. Over-expression of APX was associated with increased APX activity in spermidine-treated seedlings whereas the addition of DMTU to spermidine completely prevented spermidine-induced increase of APX activity. Thus, DMTU permitted the demonstration that exogenous spermidine supplied to maize seedlings causes an oxidative stress and induces APX, a key enzyme of the antioxidant defence mechanism, through H₂O₂, a spermidine catabolic product.     

Aluminum inhibits root growth and induces hydrogen peroxide accumulation in Plantago algarbiensis and P. almogravensis seedlings

We have evaluated the impact of aluminum (Al) on germination, relative root growth, Al accumulation in roots tips, H₂O₂ levels, plasma membrane integrity, pigment levels, protein content, and the activities of superoxide dismutase (SOD) and catalase (CAT) in seedlings of the endangered Portuguese species Plantago algarbiensis and Plantago almogravensis. We found that up to 400 μM Al had no impact on the germination percentage in either species but inhibited root growth in a concentration-dependent manner (more severely in P. algarbiensis). Al accumulation in the root tips of both species was concentration dependent up to 200 μM but declined thereafter despite the absence of membrane damage. We observed a concentration-dependent induction of SOD activity but no change in CAT activity resulting in the accumulation of H₂O₂ (a known growth inhibitor), although its impact in P. almogravensis may be partially ameliorated by the accumulation of carotenoid pigments. Our data suggest an association between Al uptake, H₂O₂ production, and the inhibition of root growth during early seedling development in P. algarbiensis and P. almogravensis, although the latter is more tolerant towards higher concentrations of the metal.