Nanoscale Sulfur Improves Plant Growth and Reduces Arsenic Toxicity and Accumulation in Rice (Oryza sativa L.).

Rice is known to accumulate arsenic (As) in its grains, posing serious health concerns for billions of people globally. We studied the effect of nanoscale sulfur (NS) on rice seedlings and mature plants under As stress. NS application caused a 40% increase in seedling biomass and a 26% increase in seed yield of mature plants compared to untreated control plants. AsIII exposure caused severe toxicity to rice; however, coexposure of plants to AsIII and NS alleviated As toxicity, and growth was significantly improved. Rice seedlings treated with AsIII + NS produced 159 and 248% more shoot and root biomass, respectively, compared to plants exposed to AsIII alone. Further, AsIII + NS-treated seedlings accumulated 32 and 11% less As in root and shoot tissues, respectively, than the AsIII-alone treatment. Mature plants treated with AsIII + NS produced 76, 110, and 108% more dry shoot biomass, seed number, and seed yield, respectively, and accumulated 69, 38, 18, and 54% less total As in the root, shoot, flag leaves, and grains, respectively, compared to AsIII-alone-treated plants. A similar trend was observed in seedlings treated with AsV and NS. The ability of sulfur (S) to alleviate As toxicity and accumulation is clearly size dependent as NS could effectively reduce bioavailability and accumulation of As in rice via modulating the gene expression activity of As transport, S assimilatory, and glutathione synthesis pathways to facilitate AsIII detoxification. These results have significant environmental implications as NS application in agriculture has the potential to decrease As in the food chain and simultaneously enable crops to grow and produce higher yields on marginal and contaminated lands.

Root system architecture, physiological analysis and dynamic transcriptomics unravel the drought-responsive traits in rice genotypes.

Drought is the major abiotic factors that limit crop productivity worldwide. To withstand stress conditions, plants alter numerous mechanisms for adaption and tolerance. Therefore, in the present study, 106 rice varieties were screened for drought tolerance phenotype via exposing different concentrations of polyethylene glycol 6000 (PEG) in the hydroponic nutrient medium at the time interval of 1, 3, and 7 days to evaluate the changes in their root system architecture. Further, based on root phenotype obtained after PEG-induced drought, two contrasting varieties drought-tolerant Heena and -sensitive Kiran were selected to study transcriptional and physiological alterations at the same stress durations. Physiological parameters (photosynthesis rate, stomatal conductance, transpiration), and non-enzymatic antioxidants (carotenoids, anthocyanins, total phenol content) production indicated better performance of Heena than Kiran. Comparatively higher accumulation of carotenoid and anthocyanin content and the increased photosynthetic rate was also observed in Heena. Root morphology (length, numbers of root hairs, seminal roots and adventitious roots) and anatomical data (lignin deposition, xylem area) enable tolerant variety Heena to better maintain membrane integrity and relative water content, which also contribute to comparatively higher biomass accumulation in Heena under drought. In transcriptome profiling, significant drought stress-associated differentially expressed genes (DEGs) were identified in both the varieties. A total of 1033 and 936 uniquely upregulated DEGs were found in Heena and Kiran respectively. The significant modulation of DEGs that were mainly associated with phytohormone signaling, stress-responsive genes (LEA, DREB), transcription factors (TFs) (AP2/ERF, MYB, WRKY, bHLH), and genes involved in photosynthesis and antioxidative mechanisms indicate better adaptive nature of Heena in stress tolerance. Additionally, the QTL-mapping analysis showed a very high number of DEGs associated with drought stress at AQHP069 QTL in Heena in comparison to Kiran which further distinguishes the drought-responsive traits at the chromosomal level in both the contrasting varieties. Overall, results support the higher capability of Heena over Kiran variety to induce numerous genes along with the development of better root architecture to endure drought stress.

Bioremediation of arsenic by soil methylating fungi: Role of Humicola sp. strain 2WS1 in amelioration of arsenic phytotoxicity in Bacopa monnieri L.

Fungi mediated arsenic (As) stress modulation has emerged as an important strategy for the mitigation of As mediated stress management in plants for reducing As contamination to the food chain. In the present study, total of 45 fungal strains were isolated from the three As contaminated sites of West Bengal, India. These strains were morphologically different and inhibited variable As tolerance (10 to 5000 mg l-1As). Total 21 fungal isolates, tolerant up to 5000 mg l-1 AsV, were investigated for As removal (10 mg l-1 As) after 21 d of cultivation under laboratory conditions. The As bioaccumulation in fungal biomass ranged between 0.146 to 11.36 g kg-1 biomass. Range of volatilized As was between 0.05 to 53.39 mg kg-1 biomass. Most promising bioaccumulation and biovolatilization potential were observed in strains viz., 2WS1, 3WS1 and 2WS9. Strain 2WS1 showed highest As biovolatilization (53.39 mg kg-1 biomass) and was identified as Humicola sp. using ITS/5.8S rDNA gene sequencing. This is the first report of Humicola sp. having As biomethylation property. Best first 8 As biomethylating fungal strains were further tested for their As remediation and PGP potential in Bacopa monnieri plant grown in As contaminated soil (20 mg kg-1) in a pot experiment under greenhouse conditions. The highest leaf stem ratio and lowest As content in leaf tissues were observed in 2WS1 inoculated Bacopa monnieri plants. The presence of arsM gene in 2WS1 strain suggests As biovolatilization as possible bioremediation and As stress mitigation strategy of 2WS1. Therefore, application of this strain of Humicola sp. strain 2WS1 in As contaminated soils could be a potential and realistic mitigation strategy for reducing As contamination to cropping system coupled with enhanced productivity.

The Journey of Arsenic from Soil to Grain in Rice.

Arsenic (As) is a non-essential toxic metalloid whose elevated concentration in rice grains is a serious issue both for rice yield and quality, and for human health. The rice-As interactions, hence, have been studied extensively in past few decades. A deep understanding of factors influencing As uptake and transport from soil to grains can be helpful to tackle this issue so as to minimize grain As levels. As uptake at the root surface by rice plants depends on factors like iron plaque and radial oxygen loss. There is involvement of a number of transporters viz., phosphate transporters and aquaglyceroporins in the uptake and transport of different As species and in the movement to subcellular compartments. These processes are also affected by sulfur availability and consequently on the level of thiol (-SH)-containing As binding peptides viz., glutathione (GSH) and phytochelatins (PCs). Further, the role of phloem in As movement to the grains is also suggested. This review presents a detailed map of journey of As from soil to the grains. The implications for the utilization of available knowledge in minimizing As in rice grains are presented.

Arsenic tolerant Trichoderma sp. reduces arsenic induced stress in chickpea (Cicer arietinum).

Toxic metalloids including arsenic (As) can neither be eliminated nor destroyed from environment; however, they can be converted from toxic to less/non-toxic forms. The form of As species and their concentration determines its toxicity in plants. Therefore, the microbe mediated biotransformation of As is crucial for its plant uptake and toxicity. In the present study the role of As tolerant Trichoderma in modulating As toxicity in chickpea plants was explored. Chickpea plants grown in arsenate spiked soil under green house conditions were inoculated with two plant growth promoting Trichoderma strains, M-35 (As tolerant) and PPLF-28 (As sensitive). Total As concentration in chickpea tissue was comparable in both the Trichoderma treatments, however, differences in levels of organic and inorganic As (iAs) species were observed. The shift in iAs to organic As species ratio in tolerant Trichoderma treatment correlated with enhanced plant growth and nutrient content. Arsenic stress amelioration in tolerant Trichoderma treatment was also evident through rhizospheric microbial community and anatomical studies of the stem morphology. Down regulation of abiotic stress responsive genes (MIPS, PGIP, CGG) in tolerant Trichoderma + As treatment as compared to As alone and sensitive Trichoderma + As treatment also revealed that tolerant strain enhanced the plant's potential to cope with As stress as compared to sensitive one. Considering the bioremediation and plant growth promotion potential, the tolerant Trichoderma may appear promising for its utilization in As affected fields for enhancing agricultural productivity.

Natural variations in expression of regulatory and detoxification related genes under limiting phosphate and arsenate stress in Arabidopsis thaliana.

Abiotic stress including nutrient deficiency and heavy metal toxicity severely affects plant growth, development, and productivity. Genetic variations within and in between species are one of the important factors in establishing interactions and responses of plants with the environment. In the recent past, natural variations in Arabidopsis thaliana have been used to understand plant development and response toward different stresses at genetic level. Phosphorus deficiency negatively affects plant growth and metabolism and modulates expression of the genes involved in Pi homeostasis. Arsenate, As(V), a chemical analog of Pi, is taken up by the plants via phosphate transport system. Studies suggest that during Pi deficiency, enhanced As(V) uptake leads to increased toxicity in plants. Here, the natural variations in Arabidopsis have been utilized to study the As(V) stress response under limiting Pi condition. The primary root length was compared to identify differential response of three Arabidopsis accessions (Col-0, Sij-1, and Slavi-1) under limiting Pi and As(V) stress. To study the molecular mechanisms responsible for the differential response, comprehensive expression profiling of the genes involved in uptake, detoxification, and regulatory mechanisms was carried out. Analysis suggests genetic variation-dependent regulatory mechanisms may affect differential response of Arabidopsis natural variants toward As(V) stress under limiting Pi condition. Therefore, it is hypothesized that detailed analysis of the natural variations under multiple stress conditions might help in the better understanding of the biological processes involved in stress tolerance and adaptation.

Salicylic acid modulates arsenic toxicity by reducing its root to shoot translocation in rice (Oryza sativa L.).

Arsenic (As) is posing serious health concerns in South East Asia where rice, an efficient accumulator of As, is prominent crop. Salicylic acid (SA) is an important signaling molecule and plays a crucial role in resistance against biotic and abiotic stress in plants. In present study, ameliorative effect of SA against arsenate (As(V)) toxicity has been investigated in rice (Oryza sativa L.). Arsenate stress hampered the plant growth in terms of root, shoots length, and biomass as well as it enhanced the level of H2O2 and MDA in dose dependent manner in shoot. Exogenous application of SA, reverted the growth, and oxidative stress caused by As(V) and significantly decreased As translocation to the shoots. Level of As in shoot was positively correlated with the expression of OsLsi2, efflux transporter responsible for root to shoot translocation of As in the form of arsenite (As(III)). SA also overcame As(V) induced oxidative stress and modulated the activities of antioxidant enzymes in a differential manner in shoots. As treatment hampered the translocation of Fe in the shoot which was compensated by the SA treatment. The level of Fe in root and shoot was positively correlated with the transcript level of transporters responsible for the accumulation of Fe, OsNRAMP5, and OsFRDL1, in the root and shoot, respectively. Co-application of SA was more effective than pre-treatment for reducing As accumulation as well as imposed toxicity.

Trichoderma inoculation augments grain amino acids and mineral nutrients by modulating arsenic speciation and accumulation in chickpea (Cicer arietinum L.).

Trichoderma reesei is an industrially important fungi which also imparts stress tolerance and plant growth promotion in various crops. Arsenic (As) contamination of field soils is one of the challenging problems in agriculture, posing potential threats for both human health and the environment. Plants in association with microbes are a liable method to improve metal tolerance and enhance crop productivity. Chickpea (Cicer arietinum L.), is an important grain legume providing cheap source of protein in semi-arid regions including As affected areas. In this study we report the role of T. reesei NBRI 0716 (NBRI 0716) in supporting chickpea growth and improving soil quality in As simulated conditions. NBRI 0716 modulated the As speciation and its availability to improve grain yield and quality (amino acids and mineral content) in chickpea (C. arietinum L.) plants grown in As spiked soil (100 mg As kg(-1) soil). Arsenic accumulation and speciation results indicate that arsenate [As(V)] was the dominant species in chickpea seeds and rhizosphere soil. The Trichoderma reduced total grain inorganic As (Asi) by 66% and enhanced dimethylarsonic acid (DMA) and monomethylarsinic acid (MMA) content of seed and rhizosphere soil. The results indicate a probable role of NBRI 0716 in As methylation as the possible mechanism for maneuvering As stress in chickpea. Analysis of functional diversity using carbon source utilization (Biolog) showed significant difference in diversity and evenness indices among the soil microbial rhizosphere communities. Microbial diversity loss caused by As were prevented in the presence of Trichoderma NBRI 0716.

Comparative Transcriptional Profiling of Contrasting Rice Genotypes Shows Expression Differences during Arsenic Stress.

Accumulation of arsenic (As) in rice (Oryza sativa L.) grain is a serious concern worldwide. Long-term exposure to As affects nutritional status in rice grain and is associated with higher rates of skin, bladder, and lung cancers, and heart disease. Genotypic variations in rice for As accumulation or tolerance are prevalent and are regulated by genetic and environmental factors. To understand molecular networks involved in As accumulation, genome-wide expression analysis was performed in roots of low- and high-As accumulating rice genotypes (LARGs and HARGs). Six rice genotypes with contrasting As accumulation potential and tolerance were used in this study. Genome-wide expression analysis suggested their differential response against As stress. This study suggests up- and downregulation of a number of unique genes involved in various pathways and biological processes in response to As stress in rice genotypes. A comparison of gene expression profiles, principal component analysis, and K-means clustering suggests that an independent pathway is operating during As stress tolerance or accumulation in contrasting genotypes. It was also observed that the differential behavior of aus genotype, Nayanmoni, from other LARGs might be due to its different genetic background. Cis-motif profiling of As-induced coexpressed genes in diverse rice genotypes led to the identification of unique cis-motifs present in differentially expressed genes. This study suggests that the genetic mechanism regulating the differential As accumulation in different genotypes may not be dependent on gene expression at the transcriptional level. However, many genes identified in this study can be analyzed and used for marker-trait associations related to As accumulation in diverse genotypes around the world.

Nitric Oxide Alleviated Arsenic Toxicity by Modulation of Antioxidants and Thiol Metabolism in Rice (Oryza sativa L.).

Nitric oxide (NO) is a gaseous signaling molecule and has a profound impact on plant growth and development. It is reported to serve as pro oxidant as well as antioxidant in plant system. In the present study, we evaluated the protective role of NO against arsenate (As(V)) toxicity in rice plants. As(V) exposure has hampered the plant growth, reduced the chlorophyll content, and enhanced the oxidative stress, while the exogenous NO supplementation has reverted these symptoms. NO supplementation has reduced the arsenic (As) accumulation in root as well as shoot. NO supplementation to As(V) exposed plants has reduced the gene expression level of OsLsi1 and OsLsi2. As(V) stress significantly impacted thiol metabolism, it reduced GSH content and GSH/GSSG ratio, and enhanced the level of PCs. NO supplementation maintained the GSH/GSSG ratio and reduced the level of PCs. NO supplementation reverted As(V) induced iron deficiency in shoot and had significant impact of gene expression level of various iron transporters (OsYSL2, OsFRDL1, OsIRT1, and OsIRO2). Conclusively, exogenous application of NO could be advantageous against As(V) toxicity and could confer the tolerance to As(V) stress in rice.

Roles for root iron plaque in sequestration and uptake of heavy metals and metalloids in aquatic and wetland plants.

Toxic metal(loid) contamination of soil and sediment poses long term risk to soil and human health through plant-human or plant-animal-human food chain pathways. Iron plaque (IP) formation is frequent in aquatic and wetland plant species and is responsible for the sequestration of various metal(loids). The presence of IP may act as a buffer or barrier and may thus enhance or reduce the uptake of potentially phytotoxic metals and metalloids by plants. If IP acts as a barrier, then low IP producing macrophytes/aquatic plants may be better accumulators of toxic metals and may find use in constructed wetlands for remediation of pollutants, while high IP forming edible plant species could be safer for human consumption. Conversely, if IP acts as a buffer for mineral nutrients and toxic elements then those cultivars may be rich in nutrients, but may also cause toxicity. However, an ecotoxicological risk is also inevitable if IP rich macrophyte roots containing heavy metals are consumed by herbivores. In this review, we summarize the current understanding about the role of IP in metal and metalloid sequestration, uptake, and transport. Furthermore, we will address the role of root IP in Oryza sativa for arsenic (As) sequestration leading to lower grain As translocation, reducing the risk of human exposure.

Synthetic phytochelatins complement a phytochelatin-deficient Arabidopsis mutant and enhance the accumulation of heavy metal(loid)s.

Phytochelatins (PCs) are naturally occurring thiol-rich peptides containing gamma (γ) peptide bonds and are well known for their metal-binding and detoxification capabilities. Whether synthetic phytochelatins (ECs) can be used as an alternative approach for enhancing the metal-binding capacity of plants has been investigated in this study. The metal-binding potential of ECs has been demonstrated in bacteria; however, no report has investigated the expression of ECs in plants. We have expressed three synthetic genes encoding ECs of different lengths in wild type (WT) Arabidopsis (Col-0 background) and a phytochelatin-deficient Arabidopsis mutant (cad1-3). After exposure to different heavy metals, the transgenic plants were examined for phenotypic changes, and metal accumulation was evaluated. The expression of EC genes rescued the sensitive phenotype of the cad1-3 mutant under heavy metal(loid) stress. Transgenic Arabidopsis plants expressing EC genes accumulated a significantly enhanced level of heavy metal(loid)s in comparison with the WT plant. The mutant complementation and enhanced heavy metal(loid) accumulation in the transgenic Arabidopsis plants suggest that ECs work in a manner similar to that of PCs in plants and that ECs could be used as an alternative for phytoremediation of heavy metal(loid) exposure.

Trichoderma inoculation ameliorates arsenic induced phytotoxic changes in gene expression and stem anatomy of chickpea (Cicer arietinum).

Arsenic, a carcinogenic metalloid severely affects plant growth in contaminated areas. Present study shows role of Trichoderma reesei NBRI 0716 (NBRI 0716) in ameliorating arsenic (As) stress on chickpea under greenhouse conditions. Arsenic stress adversely affected seed germination (25%), chlorophyll content (44%) and almost eliminated nodule formation that were significantly restored on NBRI 0716 inoculation. It also restored stem anomalies like reduced trichome turgidity and density, deformation in collenchymatous and sclerenchymatous cells induced by As stress. Semi-quantitative RT-PCR of stress responsive genes showed differential expression of genes involved in synthesis of cell wall degrading enzymes, dormancy termination and abiotic stress. Upregulation of drought responsive genes (DRE, EREBP, T6PS, MIPS, and PGIP), enhanced proline content and shrunken cortex cells in the presence of As suggests that it creates water deficiency in plants and these responses were modulated by NBRI 0716 which provides a protective role. NBRI0716 mediated production of As reductase enzyme in chickpea and thus contributed in As metabolism. The study suggests a multifarious role of NBRI0716 in mediating stress tolerance in chickpea towards As.

Transcriptomic and metabolomic shifts in rice roots in response to Cr (VI) stress.

BACKGROUND: Widespread use of chromium (Cr) contaminated fields due to careless and inappropriate management practices of effluent discharge, mostly from industries related to metallurgy, electroplating, production of paints and pigments, tanning, and wood preservation elevates its concentration in surface soil and eventually into rice plants and grains. In spite of many previous studies having been conducted on the effects of chromium stress, the precise molecular mechanisms related to both the effects of chromium phytotoxicity, the defense reactions of plants against chromium exposure as well as translocation and accumulation in rice remain poorly understood. RESULTS: Detailed analysis of genome-wide transcriptome profiling in rice root is reported here, following Cr-plant interaction. Such studies are important for the identification of genes responsible for tolerance, accumulation and defense response in plants with respect to Cr stress. Rice root metabolome analysis was also carried out to relate differential transcriptome data to biological processes affected by Cr (VI) stress in rice. To check whether the Cr-specific motifs were indeed significantly over represented in the promoter regions of Cr-responsive genes, occurrence of these motifs in whole genome sequence was carried out. In the background of whole genome, the lift value for these 14 and 13 motifs was significantly high in the test dataset. Though no functional role has been assigned to any of the motifs, but all of these are present as promoter motifs in the Database of orthologus promoters. CONCLUSION: These findings clearly suggest that a complex network of regulatory pathways modulates Cr-response of rice. The integrated matrix of both transcriptome and metabolome data after suitable normalization and initial calculations provided us a visual picture of the correlations between components. Predominance of different motifs in the subsets of genes suggests the involvement of motif-specific transcription modulating proteins in Cr stress response of rice.

Effect of arsenic on growth, oxidative stress, and antioxidant system in rice seedlings.

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.

Thiol metabolism and antioxidant systems complement each other during arsenate detoxification in Ceratophyllum demersum L.

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 (As(V); 0-250 microM) exposure to explore the underlying mechanisms of As detoxification. Plants efficiently tolerated As toxicity up to concentrations of 50 microM As(V) and durations of 4 d with no significant effect on growth by modulating various pathways in a coordinated and complementary manner and accumulated about 76 microg As g(-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 As(V) through its reduction and subsequent complexation. The maximum proportion of As chelated by PCs was found to be about 30% (at 50 microM As(V) 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 50 microM As(V) for 4 d. Exposure of plants to higher concentrations (250 microM As(V)) and/or for longer durations (7 d) resulted in a significant increase in the level of As (maximum 525 microgg(-1)dw at 250 microM 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.

Arsenic hazards: strategies for tolerance and remediation by plants.

Arsenic toxicity has become a global concern owing to the ever-increasing contamination of water, soil and crops in many regions of the world. To limit the detrimental impact of arsenic compounds, efficient strategies such as phytoremediation are required. Suitable plants include arsenic hyperaccumulating ferns and aquatic plants that are capable of completing their life cycle in the presence of high levels of arsenic through the concerted action of arsenate reduction to arsenite, arsenite complexation, and vacuolar compartmentalization of complexed or inorganic arsenic. Tolerance can also be conferred by lowering arsenic uptake by suppression of phosphate transport activity, a major pathway for arsenate entry. In many unicellular organisms, arsenic tolerance is based on the active removal of cytosolic arsenite while limiting the uptake of arsenate. Recent molecular studies have revealed many of the gene products involved in these processes, providing the tools to improve crop species and to optimize phytoremediation; however, so far only single genes have been manipulated, which has limited progress. We will discuss recent advances and their potential applications, particularly in the context of multigenic engineering approaches.

Copper-induced oxidative stress and responses of antioxidants and phytochelatins in Hydrilla verticillata (L.f.) Royle.

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.

Synthesis of phytochelatins and modulation of antioxidants in response to cadmium stress in Cuscuta reflexa--an angiospermic parasite.

Effect of cadmium on growth, antioxidative enzymes namely catalase, peroxidase, glutathione reductase, level of glutathione and phytochelatin synthesis was investigated in callus and seedlings of Cuscuta reflexa. A time, concentration and tissue dependent response of Cd was observed. Cd inhibited the growth of callus and seedlings by 50% at 300 and 500 micromol/L concentrations, respectively. Shorter exposure of low concentration of Cd led to augmentation of antioxidant activity, both in callus and seedlings, while longer exposure and high concentration of Cd led to a concentration dependent decrease in callus. Analysis of phytochelatin (PC) synthesis in callus and seedlings of C. reflexa revealed both quantitative and qualitative changes. Cd at low concentrations led to synthesis of predominantly PC4, while at higher concentrations, PC3 was the major form being synthesized. Amelioration of antioxidative systems of C. reflexa in response to Cd stress might be playing a protective role, alleviating the damaging effects of ROS, generated during Cd stress. Concomitantly, chelation and sequestering of toxic Cd ions in this parasite was mediated by synthesis of PC. The response to Cd stress shown by this holoparasitic plant was found to be similar to those of non-parasitic plants (hosts).

Impacts of fly-ash on soil and plant responses.

Coal combustion produces carbon dioxides, SO x, NO x and a variety of byproducts, including fly-ash, flue gas and scrubber sludge. Fly-ash consists of minute glass-like particles and its deposition on leaves inhibits the normal transpiration and photosynthesis of plants. Fly-ash also affects the physicochemical characteristics of soil because it is generally very basic, rich in various essential and non-essential elements, but poor in both nitrogen and available phosphorus. The massive fly-ash materials have been a potential resource for the agricultural activities as well as the other industrial purposes. Practical value of fly-ash in agriculture as an 'effective and safe' fertiliser or soil amendment can be established after repeated field experiments. Here remains to be disclosed the biological processes and interactions due to 'lack and excess' of the fly-ash exposures along with abiotic and biotic factors. These may involve the symbiotic fixation of nitrogen and the biological extraction of metals following immobilisation of toxic heavy metal ions, as well as other neutralisation and equilibration processes during weathering. Nitrogen-fixing plants with an apparent heavy metal-tolerance can be helpful as the early colonisers of fly-ash dumps and nearby areas.