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Copper, though essential, is potentially toxic heavy metal at supraoptimal level and has widespread contamination. The present investigation was carried out to study the responses induced by lower as well as higher doses of copper (0.1-25 microM) in an aquatic macrophyte, Hydrilla verticillata (L.f.) Royle for a period of 1-7 days. The plants accumulated copper in high amount with a maximum of 770 microg g(-1) dw on day 7 at 25 microM. Biomass and photosynthetic pigments showed less alteration up to 1 microM while at higher concentrations, significant decline occurred. Malondialdehyde (MDA) content and electrical conductivity (EC) also showed sharp increase at higher concentrations indicating oxidative stress. In response to copper exposure, plants showed significant induction of proteins and enzymes like superoxide dismutase (SOD), ascorbate peroxidase (APX), guaiacol peroxidase (GPX), catalase (CAT) and glutathione reductase (GR), however, only up to moderate exposures. Total non-protein thiols (NP-SH) and cysteine levels increased significantly up to 5 microM copper exposure while at 25 microM, their level declined drastically. Reduced glutathione (GSH) showed decrease at all concentrations while oxidized glutathione (GSSG) simultaneously increased. Phytochelatins (PCs) were also induced significantly at studied concentrations of 1 and 5 microM on day 4 in comparison to control. However, copper chelation depicted by PC-SH to copper ratio was found to be low (6.5% at 1 microM and 2.4% at 5 microM) suggesting that PCs play only a part in integrated mechanisms of copper homeostasis and detoxification. Tolerant response of plants to moderate copper exposures and high accumulation potential warrants their suitability for remediation of moderately copper polluted water bodies.

Chloroplast-localized adenosine-5'-phosphosulphate reductase (APR) generates sulfite and plays a pivotal role in reduction of sulfate to cysteine. The peroxisome-localized sulfite oxidase (SO) oxidizes excess sulfite to sulfate. Arabidopsis wild type, SO RNA-interference (SO Ri) and SO overexpression (SO OE) transgenic lines infiltrated with sulfite showed increased water loss in SO Ri plants, and smaller stomatal apertures in SO OE plants compared with wild-type plants. Sulfite application also limited sulfate and abscisic acid-induced stomatal closure in wild type and SO Ri. The increases in APR activity in response to sulfite infiltration into wild type and SO Ri leaves resulted in an increase in endogenous sulfite, indicating that APR has an important role in sulfite-induced increases in stomatal aperture. Sulfite-induced H2O2 generation by NADPH oxidase led to enhanced APR expression and sulfite production. Suppression of APR by inhibiting NADPH oxidase and glutathione reductase2 (GR2), or mutation in APR2 or GR2 , resulted in a decrease in sulfite production and stomatal apertures. The importance of APR and SO and the significance of sulfite concentrations in water loss were further demonstrated during rapid, harsh drought stress in root-detached wild-type, gr2 and SO transgenic plants. Our results demonstrate the role of SO in sulfite homeostasis in relation to water consumption in well-watered plants. [ABSTRACT FROM AUTHOR]

Arsenic (As) is a non-essential element whose entry into rice grains is an issue of public concern. The need, therefore, exists to understand the regulatory mechanisms of As accumulation and distribution patterns in plants. This study analyzed the effect of sulfur (S) supply on As accumulation and distribution in rice ( Oryza sativa L. var. IR64) plants. Nine day old seedlings were grown in 0.798 mM S (Normal S), 0.2 mM S (Low S) and 0.003 mM S (Zero S) for 7 days and then subjected to AsIII (20 μM) exposure for 7–15 days. The concentration of As varied significantly in different S treatments with a consistent trend of decrease in As concentration in depleted S supply treatments on both 7 days and 15 days. Arsenic concentration declined from 726 μg g − 1 DW to 537 μg g − 1 DW in roots and from 29 μg g − 1 DW to 8 μg g − 1 DW in shoots at 7 days, and from 1536 μg g − 1 DW to 1062 in roots and from 58 μg g − 1 DW to 16 μg g − 1 DW in shoots at 15 days, when S supply declined from normal to zero. The subcellular distribution of As was also found to vary with alteration in S supply and also differentially in shoot and root. In general, an up-regulation of sulfate transporters of groups 1 and 2 was observed while As transporters (Lsi1 and Lsi2) were down-regulated in response to As exposure to maintain sulfate and to regulate As levels. In spite of S depletion, an increase in cysteine, glutathione (GSH) and phytochelatin (PC) levels was observed upon As exposure. However, some negative impact of S depletion and As could be seen on the growth of plants. The study concluded that even in conditions of decreased S availability, plants continue to rely on thiol metabolism to tackle As levels and its toxicity effectively and altered subcellular distribution of As contributes only partially. [ABSTRACT FROM AUTHOR]

Widespread contamination of arsenic (As) is recognized as a global problem due to its well-known accumulation by edible and medicinal plants and associated health risks for the humans. In this study, phytotoxicity imposed upon exposure to arsenate [As(V); 0-250 μM for 1-7 days] and ensuing biochemical responses were investigated in a medicinal herb Bacopa monnieri L. vis-à-vis As accumulation. Plants accumulated substantial amount of As (total 768 μg g−1 dw at 250 μM As(V) after 7 days) with the maximum As retention being in roots (60%) followed by stem (23%) and leaves (17%). The level of cysteine and total nonprotein thiols (NP-SH) increased significantly at all exposure concentrations and durations. Besides, the level of metalloid binding ligands viz., glutathione (GSH) and phytochelatins (PCs) increased significantly at the studied concentrations [50 and 250 μM As(V)] in both roots and leaves. The activities of various enzymes viz., arsenate reductase (AR), glutathione reductase (GR), superoxide dismutase (SOD), guaiacol peroxidase (GPX), ascorbate peroxidase (APX), and catalase (CAT) showed differential but coordinated stimulation in leaves and roots to help plants combat As toxicity up to moderate exposure concentrations (50 μM). However, beyond 50 μM, biomass production was found to decrease along with photosynthetic pigments and total soluble proteins, whereas lipid peroxidation increased. In conclusion, As accumulation potential of Bacopa may warrant its use as a phytoremediator but if Bacopa growing in contaminated areas is consumed by humans, it may prove to be toxic for health. © 2011 Wiley Periodicals, Inc. Environ Toxicol 28: 419-430, 2013. [ABSTRACT FROM AUTHOR]

Abstract: Thiourea (TU) has been found to enhance the stress tolerance of plants in our earlier field trials. In the present study, the TU mediated effect on the redox and antioxidant responses were studied in response to salinity (NaCl) stress in Indian mustard (Brassica juncea (L.) Czern.) seedlings. Biochemical analyses of reactive oxygen species (ROS) and lipid peroxidation revealed that TU supplementation to NaCl brought down their levels to near control values as compared to that of NaCl stress. These positive effects could be correlated to the significant increases in the 1,1-diphenyl-2-picrylhydrazyl (DPPH)-radical scavenging activity, in the levels of reduced glutathione (GSH) and GSH/GSSG (reduced/oxidized glutathione) ratio and in the activities of superoxide dismutase (SOD; EC 1.1.5.1.1) and glutathione reductase (GR; EC 1.6.4.2) in NaCl+TU treatment as compared to that of NaCl treatment. Further, TU supplementation allowed plants to avoid an over-accumulation of pyridine nucleotides, to stimulate alternative pathways (through higher glycolate oxidase activity; EC 1.1.3.15) for channeling reducing equivalents and thus, to maintain the redox state to near control levels. These positive responses were also linked to an increased energy utilization (analyzed in terms of ATP/ADP ratio) and presumably to an early signaling of the stress through stimulated activity of ascorbate oxidase (EC 1.10.3.3), an important component of stress signaling. A significant reduction observed in the level of sodium ion (Na+) accumulation indicated that TU mediated tolerance is attributable to salt avoidance. Thus, the present study suggested that TU treatment regulated redox and antioxidant machinery to reduce the NaCl-induced oxidative stress. [Copyright &y& Elsevier]

enic (As) contamination of the environment has emerged as a concerning issue recently for which phytoremediation has been suggested as a viable solution. Hydrilla verticillata (L.f.) Royle is a widely distributed rapidly growing aquatic weed possessing significant potential to accumulate As and is thus a potential candidate for the purpose of As phytoremediation. In the present study, an investigation of thiol metabolism was conducted in H. verticillata, which revealed differential effects upon exposure to arsenite [As(III)] and arsenate [As(V)]. The accumulation of arsenic was found to be higher upon exposure to As(III) than to As(V). Besides, As(III) was found to induce the activities of enzymes, such as cysteine synthase and γ-glutamylcysteine synthetase and the amounts of cysteine and glutathione (GSH) to higher levels than that observed with As(V). The activity of glutathione-S-transferase was, however, stimulated to a higher level upon exposure to As(V) than As(III). The activity of arsenate reductase was found to increase upon As(V) exposure at all concentrations and durations. In addition, a significant stimulation in the activity of phytochelatin synthase was noticed in vitro with an increase in As/GSH concentration and time of incubation. Arsenic detoxification in H. verticillata thus appeared to involve an induction of thiol synthesis and consumption in a coordinated manner, though differentially upon exposure to As(III) and As(V). The information gained through this study would help in better designing of the pilot experiment at the field level depending on the chemical composition of the contaminated water. [ABSTRACT FROM AUTHOR]

In the present study, Hydrilla verticillata plants were exposed to arsenate (AsV; 50μM) and arsenite (AsIII; 5μM) under variable S supply: deficient (2μM S, –S), normal (1mM S, +S) and excess (2mM S, +HS). Arsenic accumulation (μgg−1 dw) in +HS plants was about 2-fold higher upon exposure to both AsV (30) and AsIII (50) than that observed in +S (12 & 24) and –S (14 & 26) plants. Despite lower As accumulation, –S plants experienced the maximum oxidative stress owing to an inadequate response of enzymatic and molecular antioxidants and significant decline in total thiols and the ratio of reduced glutathione (GSH) to oxidized glutathione (GSSG). By contrast +HS plants had significant increase in total thiols and an improved redox status, did not demonstrate any negative impact to antioxidants except catalase and hence experienced the least increase in oxidative stress parameters. In conclusion, an increase in S supply to plants may improve their accumulation capacity for As through enhanced tolerance caused by a positive effect on thiol metabolism and antioxidant status of the plants. [Copyright &y& Elsevier]

The physiological, biochemical, and proteomic changes in germinating rice seedlings were investigated under arsenic stress. A marked decrease in germination percentage, shoot, and root elongation as well as plant biomass was observed with arsenic treatments, as compared to control, whereas accumulation of arsenic and malondialdehyde (MDA) in seedlings were increased significantly with increasing arsenic concentration (both AsIII and AsV). The up-regulation of some antioxidant enzyme activities and the isozymes of superoxide dismutase (SOD, EC 1.15.1.1), ascorbate peroxidase (APX, EC 1.11.1.11), peroxidase (POD, EC 1.11.1.7), and glutathione reductase (GR, 1.6.4.2) substantiated that arsenic accumulation generated oxidative stress, which was more pronounced in As(III) treatment. We also studied the protective effect of reduced glutathione (GSH) and cysteine (Cys) to As(III)/As(V) stressed seedlings. Both GSH and Cys imparted enhanced tolerance to seedlings against arsenic stress. Seedlings growth improved while level of MDA declined significantly when GSH and Cys were supplemented to As(III)/As(V) treatments suggesting GSH and Cys-mediated protection against oxidative stress. The arsenic content was highest in roots of seedlings grown in As(III) in the presence of GSH/Cys. However, in case of As(V) plus GSH or Cys, the arsenic content in seedlings was highest in shoots. The results are suggestive of differential metabolism of As(III) and As(V) in rice. [Copyright &y& Elsevier]

Abstract: Ceratophyllum demersum L. is known to be a potential accumulator of arsenic (As), but mechanisms of As detoxification have not been investigated so far. In the present study, we analyzed the biochemical responses of Ceratophyllum plants to arsenate (AsV; 0–250μM) exposure to explore the underlying mechanisms of As detoxification. Plants efficiently tolerated As toxicity up to concentrations of 50μM AsV and durations of 4 d with no significant effect on growth by modulating various pathways in a coordinated and complementary manner and accumulated about 76μgAsg−1 dw. Significant increases were observed in the levels of various thiols including phytochelatins (PCs), the activities of enzymes of thiolic metabolism as well as arsenate reductase (AR). These primary responses probably enabled plants to detoxify at least some part of AsV through its reduction and subsequent complexation. The maximum proportion of As chelated by PCs was found to be about 30% (at 50μM AsV after 2 d). Simultaneously, a significant increase in the activities of antioxidant enzymes was observed and hence plants did not experience oxidative stress when exposed to 50μM AsV for 4 d. Exposure of plants to higher concentrations (250μM AsV) and/or for longer durations (7 d) resulted in a significant increase in the level of As (maximum 525μgg−1 dw at 250μM after 7 d) and an inverse relationship between As accumulation and various detoxification strategies was observed that lead to enhanced oxidative stress and hampered growth. [Copyright &y& Elsevier]

In the present study, the level of thiols and activity of related enzymes were investigated in coontail (Ceratophyllum demersum L.) plants to analyze their role in combating the stress caused upon exposure to cadmium (Cd; 0–10μM) for a duration up to 7d. Plants showed the maximum accumulation of 1293μgCdg−1 dw after 7d at 10μM. Significant increases in the level of total non-protein thiols (NP-SH) including phytochelatins (PCs) as well as upstream metabolites of the PC biosynthetic pathway, cysteine and glutathione (GSH) were observed. In addition, significant increases in the activities of cysteine synthase (CS), glutathione-S-transferase (GST), glutathione reductase (GR), as well as in vitro activation of phytochelatin synthase (PCS), were noticed in response to Cd. In conclusion, under Cd stress, plants adapted to a new metabolic equilibrium of thiols through coordinated synthesis and consumption to combat Cd toxicity and to accumulate it. [Copyright &y& Elsevier]