Plant steroid hormones produced under Ni stress are involved in the regulation of metal uptake and oxidative stress in Brassica juncea L

Brassinosteroids (BRs) are involved in the amelioration of various biotic and abiotic stresses. With an aim to explore the role of BRs under heavy metal stress, plants of Brassica juncea L. were grown in pots. The plants were subjected to various concentrations of Nickel metal (0.0, 0.2, 0.4 and 0.6 mM) and harvested on 60th day in order to observe the expression of these hormones. The isolated BRs from the leaves of Brassica plants characterized by GC-MS include 24-Epibrassinolide (24-EBL), Castasterone, Dolicholide and Typhasterole. The effect of isolated 24-EBL was studied on Ni metal uptake and antioxidative defense system in 60 d old plants of Brassica. It was observed that 24-EBL significantly increased the activities of stress ameliorating enzymes and lowered the metal uptake in plants. This is the first report in B. juncea L. plants showing the expression of BRs under metal treatments and effect of the isolated 24-EBL on metal uptake and in oxidative stress management.     

Differential physiological,ultramorphological and metabolic responses of cotton cultivars under cadmium stress

Cadmium (Cd) stress may cause serious physiological, ultramorphological and biochemical anomalies in plants. Cd-induced physiological, subcellular and metabolic alterations in two transgenic cotton cultivars (BR001, GK30) and their parent line (Coker 312) were evaluated using 10, 100 and 1000 μM Cd. Germination, fresh biomass of roots, stems and leaves were significantly inhibited at 1000 μM Cd. Root volume tolerance index significantly increased (124.16%) in Coker 312 at 1000 μM Cd. In non-Cd stressed conditions, electron micrographs showed well-configured root meristem and leaf mesophyll cells. At 1000 μM Cd, greater ultramorphological alterations were observed in BR001 followed by GK30 and Coker 312. These changes were observed in nucleus, vacuoles, mitochondria and chloroplast. Dense precipitates, probably Cd, were seen in vacuoles, which were also attached to the cell walls. A considerable increase in number of nuclei, vacuoles, starch granules and plastoglobuli was observed in the electron micrographs of both roots and leaves at 1000 μM Cd. MDA contents were higher in roots of BR001 at 1000 μM Cd. Mean values of SOD activity in leaves of both BR001 and GK30 at 1000 μM Cd significantly increased as compared to the controls. POD activity in roots of BR001 and Coker 312 was greater at all Cd (10, 100, 1000 μM) levels over the control. Regarding APX, highest percent increase (71.64%) in roots of GK30 at 1000 μM Cd was found. Non-significant differences in CAT activity were observed at all levels of Cd stress in leaves of BR001 and GK30. Both transgenic cotton cultivars and their parental line invariably responded towards Cd stress. However, Coker 312 showed Cd-resistant behavior as compared to its progeny lines (BR001 and GK30).     

Oxidative stress related enzymes in response to chromium (Ⅵ) toxicity in Oxya chinensis ( Orthoptera : Acridoidae)

The toxic effects of Cr（Vl） on antioxidant enzymes of Oxya chinensis（Orthoptera： Acridoidae） were determined. Changes in the activities of the antioxidant enzymes superoxide dismutase（SOD）, catalase（CAT）, and guaiacol peroxidase（GPx） were measured in O. chinensis insects injected with Cr（VI）. Fifth-nymphs of O. chinensis insects were injected with Cr（VI） with different concentrations （0, 75, 150, 225, 300, 375, 450 mg/kg of body weight）. The results showed that Cr（VI） led to the change of SOD, CAT, and GPx activities at different concentrations, which revealed that： （1） The oxidative stress of SOD increased with the increase of Cr （VI） concentration. （2） With the increase of Cr （VI） concentrations, CAT activities for females increased at lower concentrations, but decreased at higher concentration range, which indicated that antioxidant system of O. chinensis was not influenced by the presence of Cr （VI）. A very similar response to Cr（VI） effect for males indicated that Cr（VI） concentrations were not high enough to damage O. chinensis in terms of CAT. （3） The GPx activity for females increased in all treatments, which revealed that the damage power of Cr（VI） was increased with the increase of Cr（Vi） concentrations in terms of GPx, but the effect was not so remarkable. There was not a consistent trend of GPx activities for males in all treatments of Cr（VI）. Cr（VI）-induced changes in antioxidant enzymes were different for SOD, CAT and GPx, of which the tendency was that activities generally changed with increase of concentrations of Cr（VI） suggesting SOD, CAT, and GPx could serve as indices of oxidative stress to some extent.     

Antioxidative responses of duckweed (<Emphasis Type="Italic">Lemna minor</Emphasis> L.) to short-term copper exposure

Goal, Scope and Background

Elevated concentrations of copper in the environment result in accumulation of the metal in plants and cause an increase in reactive oxidative species (ROS). The first response to elevated amounts of ROS is increased levels of enzymatic and non-enzymatic antioxidants that reduce oxidative stress. The aim of our study was to evaluate the early stages of antioxidative responses to the low copper concentrations usually present in moderately polluted environments. In addition, some other parameters were examined to evaluate the effect of copper on plants.

Methods

Duckweed (Lemna minor L.) was exposed to different concentrations of copper sulphate for up to 24 hours. Glutathione concentration and enzymatic activities of catalase, guaiacol peroxidase and glutathione reductase were measured spectrophotometrically. Additionally, delayed and prompt chlorophyll fluorescence was measured by luminometry and fluorometry, respectively. The accumulation of copper in plants exposed for 24 hours to various concentrations of copper sulphate was measured by flame atomic absorption spectrophotometry.

Results

The treatment of plants with copper sulphate resulted in an immediate decrease of the glutathione pool, which was replenished after 24 hours at CuSO₄ concentrations lower than 2 μM. Higher CuSO₄ concentrations caused a decrease of reduced glutathione. The responses of the antioxidant enzymes glutathione reductase, guaiacol peroxidase and catalase to CuSO₄ differed during the first six hours of exposure, but their enzyme activities all increased after 24 hours of exposure. All these enzymes displayed biphasic activity curves with maximum values between 0.5 μM and 1 μM CuSO₄. The response of guaiacol peroxidase was the most pronounced and statistically significantly specific and that of catalase the least. Delayed chlorophyll fluorescence decreased after exposure to 1 μM CuSO₄, but no significant effect on maximum quantum yield of photosystem II (Fv/Fm) was observed. L. minor accumulated relatively high concentrations of copper. The accumulation rate was higher at lower concentrations of copper in the test medium (up to 2 μM CuSO₄) than at concentrations above 2 μM CuSO₄.

Discussion

One of the most pronounced antioxidative responses to copper exposure was modified levels of oxidized and reduced forms of glutathione. The decrease of the glutathione pool is most probably coupled with induced production of phytochelatins. Antioxidative enzymes showed the biphasic enzyme activity characteristic of stress response. Guaiacol peroxidase exhibited the greatest significant increase of activity, even at higher CuSO₄ concentrations at which the activity of catalase and glutathione reductase dropped. The intensity of delayed chlorophyll fluorescence decreased, indicating reduced photosynthesis of plants under stress. All the measured parameters showed that plants respond to even low copper concentrations very soon after exposure. The accumulation rate of copper in duckweed tissues indicates that L. minor is an accumulator species.

Conclusions

The synchronized and prompt inducibility of antioxidants indicates their involvement in a general plant defence strategy for coping with metal-induced oxidative stress. Glutathione concentration and guaiacol peroxidase activity were found to be the most sensitive of the early indicators of exposure to copper concentrations present in polluted water bodies.

Recommendation and Perspectives

The experimental design of the present study allowed us to compare the sensitivity of various methods and parameters for detecting plant responses to heavy metal-induced oxidative stress. The level of glutathione and the enzyme activities of guaiacol peroxidase and glutathione reductase could be used as a rapidly determined early warning system in toxicity studies.

     

Oxidative stress related enzymes in response to chromium (Ⅵ) toxicity in Oxya chinensis ( Orthoptera : Acridoidae)

The toxic effects of Cr(Ⅵ) on antioxidant enzymes of Oxya chinensis(Orthoptera: Acridoidae) were determined. Changes in the activities of the antioxidant enzymes superoxide dismutase(SOD), catalase(CAT), and guaiacol peroxidase(GPx) were measured in O. chinensis insects injected with Cr(Ⅵ). Fifth-nymphs of O. chinensis insects were injected with Cr(Ⅵ) with different concentrations (0, 75, 150, 225, 300, 375, 450 mg/kg of body weight). The results showed that Cr(Ⅵ) led to the change of SOD, CAT, and GPx activities at different concentrations, which revealed that: (1) The oxidative stress of SOD increased with the increase of Cr (Ⅵ)concentration. (2) With the increase of Cr (Ⅵ) concentrations, CAT activities for females increased at lower concentrations, but decreased at higher concentration range, which indicated that antioxidant system of O. chinensis was not influenced by the presence of Cr (Ⅵ). A very similar response to Cr(Ⅵ) effect for males indicated that Cr(Ⅵ) concentrations were not high enough to damage O. chinensis in terms of CAT. (3) The GPx activity for females increased in all treatments, which revealed that the damage power of Cr(Ⅵ)was increased with the increase of Cr(Ⅵ) concentrations in terms of GPx, but the effect was not so remarkable. There was not a consistent trend of GPx activities for males in all treatments of Cr(Ⅵ). Cr(Ⅵ)-induced changes in antioxidant enzymes were different for SOD, CAT and GPx, of which the tendency was that activities generally changed with increase of concentrations of Cr(Ⅵ) suggesting SOD, CAT, and GPx could serve as indices of oxidative stress to some extent.     

Epibrassinolide ameliorates Cr (VI) stress via influencing the levels of indole-3-acetic acid, abscisic acid, polyamines and antioxidant system of radish seedlings

The present investigation determined the effects of epibrassinolide (EBL) on the levels of indole-3-acetic acid (IAA), abscisic acid (ABA), and polyamine (PA) and antioxidant potential of 7-d old Raphanus sativus L. cv. ‘Pusa chetki’ seedlings grown under Cr (VI) metal stress. Reduced titers of free (0.767 μg g⁻¹ FW) and bound (0.545 μg g⁻¹ FW) IAA in Cr (VI) stressed seedlings were observed over untreated control. Supplementations of EBL to Cr (VI) stressed seedlings were able to enhance both free (2.14-5.68 μg g⁻¹ FW) and bound IAA (2.45-7.78 μg g⁻¹ FW) concentrations in comparison to Cr (VI) metal treatment alone. Significant rise in free (13.49 μg g⁻¹ FW) and bound (12.17 μg g⁻¹ FW) ABA contents were noticed for Cr (VI) stressed seedlings when compared to untreated control. No significant increase in ABA contents were recorded for Cr (VI) stressed seedlings upon supplementation with EBL over Cr (VI) treatment alone. A significant increase in Put (18.40 μg g⁻¹ FW) and Cad (9.08 μg g⁻¹ FW) contents were found for 10⁻⁹ M EBL plus Cr (VI) metal treatments when compared to Cr (VI) treatment alone. Spermidine (Spd) contents were found to decline significantly for EBL treatment alone or when supplemented with Cr (VI) treatments over untreated controls and Cr (VI) treatment alone. Antioxidant levels were found to enhance, with glutathione (57.98 mg g⁻¹ FW), proline (4.97 mg g⁻¹ FW), glycinebetaine (39.01 μmol mL⁻¹), ascorbic acid (3.17 mg g⁻¹ FW) and phytochelatins (65.69 μmol g⁻¹ FW) contents noted for EBL supplemented to Cr (VI) metal solution over Cr (VI) treatment alone. Reduced activities of guaiacol peroxidase (0.391 U mg⁻¹ protein) and catalase (0.221 U mg⁻¹ protein) and enhanced activities of glutathione reductase (7.14 U mg⁻¹ protein), superoxide dismutase (15.20 U mg⁻¹ protein) and ascorbate peroxidase (4.31 U mg⁻¹ protein) were observed in seedlings treated with EBL plus Cr (VI) over Cr metal treatment alone. Reduced MDA (2.55 μmol g⁻¹ FW) and H₂O₂ (33.24 μmol g⁻¹ FW) contents were recorded for 10⁻⁹ M EBL supplemented to Cr (VI) stress over Cr (VI) treatment alone. Enhancement in free radical scavenging potential as indicated by higher values of 1,1-diphenylpicrylhydrazyl, deoxyribose and reducing power activity assays, and increased levels of phenols and soluble sugars also showed significant influence of EBL in alleviating Cr (VI) stress in radish seedlings.     

反胶束体系中漆酶催化愈创木酚的研究

采用4种不同属性类型的表面活性剂(CTAB,Tween-80, AOT以及鼠李糖脂)构建反胶束体系,以木质素模式物愈创木酚为底物,研究了漆酶在反胶束环境中催化愈创木酚的特性,通过对不同反胶束环境中漆酶酶活的对比,确定在4种反胶束体系中,适合酶解反应的最佳表面活性剂浓度和对应的最佳含水率w0.分别为:CTAB浓度20mmol/L,w0=20;Tween 80浓度50mmol/L,w0=25;AOT浓度30mmol/L,w0=10;鼠李糖脂浓度10mmol/L,w0=30.并考察了表面活性剂类型属性对愈创木酚在反胶束体系中酶解行为的影响和底物动力学.结果表明,最佳表面活性剂浓度以及对应的最适含水率条件下,阴离子表面活性剂AOT和鼠李糖脂的反胶束体系中愈创木酚底物的催化效果较好,鼠李糖脂反胶束体系中底物的催化率最高,反应6h后分别比 CTAB、Tween-80以及AOT反胶束体系中的转化率高44.12%、34.6%、29.43%.