Soil fungi for mycoremediation of arsenic pollution in agriculture soils.

AIMS: Soil arsenic (As) contamination of food-chains and public health can be mitigated through fungal bioremediation. To enumerate culturable soil fungi, soils were collected from the As-contaminated paddy fields (3-35 mg kg(-1) ) of the middle Indo-Gangetic Plains. METHODS AND RESULTS: Total 54 fungal strains were obtained and identified at their molecular level. All strains were tested for As tolerance (from 100 to 10,000 mg l(-1) arsenate). Fifteen fungal strains, tolerant to 10,000 mg l(-1) arsenate, were studied for As removal in-vivo for 21 days by cultivating them individually in potato dextrose broth enriched with 10 mg l(-1) As. The bioaccumulation of As in fungal biomass ranged from 0·023 to 0·259 g kg(-1). The biovolatilized As ranged from 0·23 to 6·4 mg kg(-1). CONCLUSIONS: Higher As bioaccumulation and biovolatilization observed in the seven fungal strains, Aspergillus oryzae FNBR_L35; Fusarium sp. FNBR_B7, FNBR_LK5 and FNBR_B3; Aspergillus nidulans FNBR_LK1; Rhizomucor variabilis sp. FNBR_B9; and Emericella sp. FNBR_BA5. These fungal strains were also tested and found suitable for significant plant growth promotion in the calendula, withania and oat plants in a greenhouse based pot experiment. SIGNIFICANCE AND IMPACT OF STUDY: These fungal strains can be used for As remediation in As-contaminated agricultural soils.

Constructed wetland as an ecotechnological tool for pollution treatment for conservation of Ganga river.

With aim to develop an efficient and ecofriendly approach for on-site treatment of sewage, a sub-surface flow constructed wetland (CW) has been developed by raising potential aquatic macrophytes; Typha latifolia, Phragmites australis, Colocasia esculenta, Polygonum hydropiper, Alternanthera sessilis and Pistia stratoites in gravel as medium. Sewage treatment potential of CW was evaluated by varying retention time at three different stages of plant growth and stabilization. After 6 months, monitoring of fully established CW indicated reduction of 90%, 65%, 78%, 84%, 76% and 86% of BOD, TSS, TDS, NO3-N, PO4-P and NH4-N, respectively in comparison to inlet after 36 h of retention time. Sewage treatment through CW also resulted in reduction of heavy metal contents. Thus, CW proved an effective method for treatment of wastewater and may be developed along river Ganga stretch as an alternative technology. Treated water may be drained into river to check further deterioration of Ganga water quality.

Investigation of arsenic accumulation and tolerance potential of Sesuvium portulacastrum (L.) L.

Sesuvium portulacastrum (L.) L., a facultative halophyte, is considered a suitable candidate for the phytoremediation of metals. An investigation of As accumulation and tolerance was conducted in Sesuvium plants upon exposure to As(V) (100-1000 µM) for 30 d. Plants demonstrated a good growth even after prolonged exposure (30 d) to high As(V) concentrations (1000 µM) and a significant As accumulation (155 µg g⁻¹ dry weight) with a bioaccumulation factor of more than ten at each concentration. The results of shoot and root dry weight, malondialdehyde accumulation, photosynthetic pigments, and total soluble proteins demonstrated that plants did not experience significant toxicity even at 1000 µM As(V) after 30 d. However, metabolites (total non-protein thiols and cysteine) and enzymes (serine acetyltransferase, cysteine synthase and γ-glutamylcysteine synthetase) of thiol metabolism, in general, remained either unaffected or showed slight decline. Hence, plants tolerated high As(V) concentrations without an involvement of thiol metabolism as a major component. Taken together, the results indicate that plants are potential As accumulator and may find application in the re-vegetation of As contaminated sites.

Arsenate exposure affects amino acids, mineral nutrient status and antioxidants in rice (Oryza sativa L.) genotypes.

Simulated pot experiments were conducted on four rice (Oryza sativa L.) genotypes (Triguna, IR-36, PNR-519, and IET-4786) to examine the effects of As(V) on amino acids and mineral nutrient status in grain along with antioxidant response to arsenic exposure. Rice genotypes responded differentially to As(V) exposure in terms of amino acids and antioxidant profiles. Total amino acid content in grains of all rice genotypes was positively correlated with arsenic accumulation. While, most of the essential amino acids increased in all cultivars except IR-36, glutamic acid and glycine increased in IET-4786 and PNR-519. The level of nonprotein thiols (NPTs) and the activities of superoxide dismutase (SOD; EC 1.15.1.1), glutathione reductase (GR; EC 1.6.4.2) and ascorbate peroxidase (APX; EC 1.11.1.11) increased in all rice cultivars except IET-4786. A significant genotypic variation was also observed in specific arsenic uptake (SAU; mg kg(-1)dw), which was in the order of Triguna (134) > IR-36 (71) > PNR-519 (53) > IET-4786 (29). Further, application of As(V) at lower doses (4 and 8 mg L(-1) As) enhanced the accumulation of selenium (Se) and other nutrients (Fe, P, Zn, and S), however, higher dose (12 mg L(-1) As) limits the nutrient uptake in rice. In conclusion, low As accumulating genotype, IET-4786, which also had significantly induced level of essential amino acids, seems suitable for cultivation in moderately As contaminated soil and would be safe for human consumption.

Differential transcriptional expression following thidiazuron-induced callus differentiation developmental shifts in rice.

Very little is known about molecular events associated with callus differentiation in indica rice. The genes expressed differentially during shoot meristem initiation were identified on genomic arrays applied to efficiently regenerating rice calli. A thidiazuron (TDZ; N-phenyl-N-thiadiazol-1,2,3-5,ylurea)-dependent regeneration protocol was developed for efficient embryogenesis in indica rice. The regenerating embryogenic calli induced by TDZ for 10 days showed transcriptional modulation of a number of genes associated with photosynthesis, hormone metabolism, plant development, signal transduction, light response, and plant defense. Eighteen candidate miRNAs were predicted to target the genes expressed differentially in the embryogenic calli grown in TDZ-containing medium. The majority of the photosynthesis-related genes up-regulated in differentiating calli were not expressed or were down-regulated in developing seeds and inflorescences. Most of the genes down-regulated in differentiating calli were up-regulated in developing seeds. The transcriptome of differentiating callus most closely resembled that of the germinating whole seed.

Arsenic accumulation pattern in 12 Indian ferns and assessing the potential of Adiantum capillus-veneris, in comparison to Pteris vittata, as arsenic hyperaccumulator.

The present study was undertaken to evaluate the ability of some Indian ferns to accumulate and tolerate arsenic. Twelve species of Indian ferns were exposed to 10 mg L(-1) arsenic as sodium arsenate for 15 days in hydroponic system. Depending on the arsenic uptake in the plant parts--Pteris vittata, Pteris cretica, Adiantum capillus-veneris and Nephrolepis exaltata may be categorised as arsenic accumulator. Further, A. capillus-veneris plants were grown in arsenic contaminated soil (200-600 mg kg(-1)) under green-house condition, to assess its arsenic accumulation and tolerance mechanism, in comparison to known As-hyperaccumulator--P. vittata Linn., growing in the same conditions. The experiment identified A. capillus-veneris having a potential to tolerate arsenic up to 500 mg kg(-1). The plants were analysed for the extent of oxidative stress, as a result of arsenic accumulation. A. capillus-veneris was able to detoxify the arsenic stress through induction of anti-oxidant defence system.

Cadmium-induced biochemical responses of Vallisneria spiralis.

The following study was carried out to investigate the cadmium (Cd) accumulating potential of Vallisneria. After subjecting plants to different concentrations of Cd, it was observed that plants are able to accumulate ample amount of metal in their roots (5,542 microg g(-1) dw) and leaves (4,368 microg g(-1) dw) in a concentration- and duration-dependent manner. Thus, it is evident that the accumulation in roots was 1.3 times higher than the shoots. It was also noted that with increasing Cd accumulation, roots of the plant appeared darker in color and harder in texture. In response to metal exposure, amount of low molecular weight antioxidants such as cysteine and nonprotein thiols (NP-SH) and activity of enzymes such as APX and GPX were significantly enhanced at lower concentrations of Cd, followed by decline at higher doses. It was also observed that in exposed plants, activity of APX enzyme was higher in roots (ca. 3 times) as compared to leaves. However, chlorophyll and protein content was found to decline significantly in a dose-dependent manner. Results suggested that due to its high accumulation potential, Vallisneria may be effectively grown in water bodies moderately contaminated with Cd.

Arsenic affects mineral nutrients in grains of various Indian rice (Oryza sativa L.) genotypes grown on arsenic-contaminated soils of West Bengal.

The exposure of paddy fields to arsenic (As) through groundwater irrigation is a serious concern that may not only lead to As accumulation to unacceptable levels but also interfere with mineral nutrients in rice grains. In the present field study, profiling of the mineral nutrients (iron (Fe), phosphorous, zinc, and selenium (Se)) was done in various rice genotypes with respect to As accumulation. A significant genotypic variation was observed in elemental retention on root Fe plaque and their accumulation in various plant parts including grains, specific As uptake (29-167 mg kg(-1) dw), as well as As transfer factor (4-45%). Grains retained the least level of As (0.7-3%) with inorganic As species being the dominant forms, while organic As species, viz., dimethylarsinic acid and monomethylarsonic acid, were non-detectable. In all tested varieties, the level of Se was low (0.05-0.12 mg kg(-1) dw), whereas that of As was high (0.4-1.68 mg kg(-1) dw), considering their safe/recommended daily intake limits, which may not warrant their human consumption. Hence, their utilization may increase the risk of arsenicosis, when grown in As-contaminated areas.

Lead bioaccumulation potential of an aquatic macrophyte Najas indica are related to antioxidant system.

Plants of Najas indica bioaccumulated significantly higher amounts of Pb (3554 microg g(-1) dw) when, exposed to varying concentrations of Pb(NO(3))(2).This also led to increased malondialdehyde (MDA), electrical conductivity (EC) and H(2)O(2) content. In response to this, the activities of antioxidant enzymes such as superoxide dismutase (SOD), ascorbate peroxidase (APX), guaiacol peroxidase (GPX), catalase (CAT) and glutathione reductase (GR) were elevated along with the induction of various molecular antioxidants including GSH, cysteine, ascorbic acid and proline. Further, Pb exposed plants showed significantly increased cysteine synthase and glutathione-S-transferase activity. Visible symptoms of toxicity were evident at 50 microM after 4d showing chlorosis and fragmentation of leaves with mucilaginous discharge. It seems that bioaccumulated Pb is efficiently tolerated by Najas plants through activation of antioxidant system and thiolic pathways which was evident by the increased biomass up to 10 microM Pb. Therefore, it appears that due to metal tolerance characteristics with high concentration factor these plants can find use in phytoremediation of aquatic system highly contaminated by Pb.

Impact of rhizobacteria on growth and chromium accumulation in Scirpus lacustris L. grown under chromium supplementation.

Four chromate tolerant rhizobacterial strains viz., RZB-01, RZB-02, RZB-03 and RZB-04 were isolated from rhizosphere of Scirpus lacustris collected from Cr-contaminated area. These strains characterized at morphological and biochemical levels. The most efficient chromate tolerant strain RZB-03 was inoculated to fresh plant of S. lacustris and grown in 2 microg ml(-1) and 5 microg ml(-1) of Cr+6 supplemented nutrient solution under controlled laboratory condition. The effects of rhizobacterial inoculation on growth and chromium accumulation in S. lacustris were evaluated. The inoculation of rhizobacteria increased biomass by 59 and 104%, while total chlorophyll content by 1.76 and 15.3% and protein content increased by 23 and 138% under 2 microg ml(-1) and 5 microg ml(-1) concentrations of Cr+6, respectively after 14 d as compared to non-inoculated plant. Similarly, the Cr accumulation also increased by 97 and 75% in shoot and 114 and 68% in root of inoculated plants as compared to non inoculated plants at 2 microg ml(-1) and 5 microg ml(-1) Cr+6 concentrations, respectively after 14 d. The chromate tolerant rhizobacteria which play an important role in chromium uptake and growth promotion in plant may be useful in development of microbes assisted phytoremediation system for decontamination of chromium polluted sites.

Characterization of native microalgal strains for their chromium bioaccumulation potential: phytoplankton response in polluted habitats.

Due to its various uses, Cr contamination has become widespread in a diverse array of environments. The present study was carried out during 2007-2008 to investigate the accumulation potential of metals (Cr, Cu, Fe, Mn, Ni and Zn) and metalloid (As) by green (GA) and blue green (BGA) microalgae growing naturally in selected Cr-contaminated sites in districts Unnao and Kanpur (Uttar Pradesh, India). This investigation is a preliminary work to identify suitable native microalgae for biomonitoring and phytoremediation purposes. A total of 22 GA and 11 BGA were encountered in three seasons (summer, rainy and winter). Among these, the accumulation potential was evaluated in high biomass producing strains of BGA (three) and GA (nine). The maximum accumulation of Cr was shown by Phormedium bohneri (8550 microg g(-1)dw) followed by Oscillatoria tenuis (7354 microg g(-1)dw), Chlamydomonas angulosa (5325 microg g(-1)dw), Ulothrix tenuissima (4564 microg g(-1)dw), and Oscillatoria nigra (1862 microg g(-1)dw); all of which demonstrated a transfer factor of >10% for Cr. The results also indicate that the phytoplankton diversity was modified by Cr pollution. BGA represented the dominant community where Cr concentration was higher (11.84 and 2.27 mg L(-1)) (r=0.695), whereas GA showed negative correlation with respect to Cr concentration (r=-0.567). In conclusion, different algal species were able to grow in Cr-contaminated sites and to accumulate significant amounts of Cr with a high transfer factor.

Antioxidant defense mechanism in hydroponically grown Zea mays seedlings under moderate lead stress.

The present study was designed to study the process of stress adaptation in roots and shoot of Zea mays seedlings grown under hydroponic conditions during exposure to lead (Pb) (0-200 microM) for 1-7 d. The alterations in growth and in the level of various biochemical parameters were accessed vis-à-vis Pb accumulation. The accumulation of Pb increased in a concentration-duration-dependent manner, however its translocation from root to shoot was low. At the same time, the level of malondialdehyde (MDA) increased with increasing Pb concentration. However, growth parameters, such as dry weight and root length did not show a significant decline to any of the Pb concentrations. In addition, the level of photosynthetic pigments decreased only upon exposure to high Pb concentrations. These results suggested an alleviation of the stress that was presumably being achieved by antioxidants viz., superoxide dismutase (SOD) and catalase (CAT) as well as ascorbic acid (AsA), which increased linearly with increasing Pb levels and exposure time. However, the level of non-protein thiols (NP-SH) in roots, in general, showed a decline beyond 4d that could be attributed to their consumption for the purpose of Pb detoxification. In conclusion, Zea mays can be used as an indicator species for Pb, and the various antioxidants might play a key role in the detoxification of Pb induced toxic effects.

Chromium (VI) induced phytotoxicity and oxidative stress in pea (Pisum sativum L.): biochemical changes and translocation of essential nutrients.

Due to widespread industrial use, chromium (Cr) is considered a hazardous environmental pollutant. It is known to inhibit plant growth and development. The present study provides the evidence of the phytotoxicity of this metal on the pea (Pisum sativum L. cv Azad) plants. The plants of pea (Pisum sativum L.) were grown in refined sand under different concentrations i.e. 0.05, 0.1, 0.2, 0.3 and 0.4 mM of Cr (VI) in order to study the effect on growth and yield, photosynthetic pigments, relative water content, non-reducing sugar and protein with activity of certain enzymes like catalase, peroxidase, starch phosphorylase and ribonuclease. The analysis of the results showed that photosynthetic pigments (68.68%), relative water contents (62.77%), non-reducing sugar (66.66%) and protein (81.57%) were decrease along with reduction in plant height (52.69%) and leaf area (50.81%) of the pea plants. However, in response to various concentration of Cr exposed plants showed significant induction of reducing and total sugars with enzymes like catalase, starch phosphorylase and ribonuclease. The translocation of Cr in various part of pea plant have been found in order of root> stem> leaves>seeds which ranged between 34.8 to 217.3 mg g(-1) d.wt. (dry weight) in roots, 6.5 to 173.13 mg g(-1) d.wt. in shoot, 4.2 to 74.43 mg g(-1) d.wt. in leaves and 0.94 to 8.64 mg g(-1) d.wt. in seeds, that is also reflected by the transfer factor of Cr from refined sand to tested species.

Thiol metabolism play significant role during cadmium detoxification by Ceratophyllum demersum L.

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; 0-10 microM) for a duration up to 7d. Plants showed the maximum accumulation of 1293 microg Cd g(-1)dw after 7d at 10 microM. 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.

Responses of cyanobacterium Anabaena doliolum during nickel stress.

Growth and biochemical responses of heterocystous nitrogen fixing cyanobacteria Anabaena doliolum were studied upon exposure to various concentrations of nickel (0.1 to 100 microM) for duration of up to 4 days, in view of its tolerance. The growth of the cyanobacterium was increased in terms of cell density (O.D), heterocyst frequency chlorophyll-a, carotenoid and c-phycocynin up to moderate exposure (10 microM for 96 hr of Ni treatment). Protein and total non protein thiol were measured as stress responsive metabolites and frequency of heterocyst and spores were observed in responses to Ni. The present study demonstrated that the tested cyanobacterium exhibited dose specific responses of metal towards studied different morphological, physiological and biochemical parameters.

Arsenic accumulation in root and shoot vis-a-vis its effects on growth and level of phytochelatins in seedlings of Cicer arietinum L.

Arsenic (As) contamination of water and soil has become a subject of prime interest due to its direct effect on human health through drinking water and food. In present study two varieties (CSG-8962 and C-235) of chickpea, Cicer arietinum L., which is a major supplementary food in many parts of India and a valuable source of protein, has been selected to estimate the level of arsenate in root and shoot of five day old seedlings vis-à-vis effect of arsenate on seedling growth and induction of thiols including glutathione (GSH) and phytochelatins (PCs) and their homologues. Both varieties accumulated arsenate to similar levels and most of the metalloid was confined to roots, only about 2.5% was translocated to shoot. Plant growth was also not affected significantly in both the varieties. Arsenate exposure significantly induced the levels of thiols including PCs and homophytochelatins (hPCs). The induction of thiols was much higher in roots than shoots and was greater in var C-235 between the two tested ones. Thus, both varieties tolerated and detoxified arsenic through chelation with GSH, PCs and hPCs, primarily in roots, however var C-235 performed better

Screening of native plants and algae growing on fly-ash affected areas near National Thermal Power Corporation, Tanda, Uttar Pradesh, India for accumulation of toxic heavy metals.

The present investigation was carried out to screen native plants growing in fly-ash (FA) contaminated areas near National Thermal Power Corporation, Tanda, Uttar Pradesh, India with a view to using them for the eco-restoration of the area. A total number of 17 plants (9 aquatic, 6 terrestrial and 2 algal species) were collected and screened for heavy metal (Fe, Zn, Cu, Mo, B, Si, Al, Cr, Pb, Cd, Hg and As) accumulation. Differential accumulation of various heavy metals by different species of plants was observed. Hydrilla verticillata was found to be the most efficient metal accumulator among 9 aquatic plants, Eclipta alba among 6 terrestrial plants and Phormedium papyraceum between 2 algal species. In general, the maximum levels of most metals were found in terrestrial plants while the lowest in algal species. However, translocation of the metals from root to shoot was found to be higher in aquatic plants than terrestrial ones. These results suggest that various aquatic, terrestrial and algal species of plants may be used in a synergistic way to remediate and restore the FA contaminated areas.

Phytoremediation efficiency of Portulaca tuberosa rox and Portulaca oleracea L. naturally growing in an industrial effluent irrigated area in Vadodra, Gujrat, India.

Phytoremediation is a novel, solar-driven and cost-effective technology for the remediation of heavy metal contaminated environments through exploitation of plants ability to accumulate heavy metals in their harvestable shoot parts. In the present investigation, we collected plants of two species of Portulaca i.e. P. tuberosa and P. oleracea from field sites in Vadodra, Gujrat, India. At one site, field was being irrigated with industrial effluent while at other with tube well water. Analysis of heavy metals was performed in industrial effluent, tube well water, soils irrigated with them, and in different parts viz., roots, stem, leaves and flowers of the plant samples. Industrial effluent and soil irrigated with it had very high level of heavy metals (Fe, Zn, Cd, Cr and As) as compared to the tube well water and soil irrigated with that. Plants of both the species growing in effluent irrigated soils showed high accumulation of metals in all plant parts with the maximum being in roots and the least in flowers. Interestingly, both species of Portulaca hyperaccumulated more than one heavy metal viz., Cd, Cr and As. The total shoot concentrations (microg g(-1) dw) of Cd, Cr and As in P. tuberosa were 1,571, 7,957 and 3,118, respectively while in P. oleracea, these were 1,128, 7,552 and 2,476, respectively. Portulaca plants have good biomass and high regeneration potential; hence appear to be suitable for the remediation of effluent (metal) contaminated areas.

Response of antioxidant enzymes in coontail (Ceratophyllum demersum L.) plants under cadmium stress.

Cadmium (Cd) contamination of aquatic systems is of major concern since it is a nonessential element and hampers plant growth upon accumulation. The aim of this study was to investigate the Cd accumulation behavior of coontail plant, Ceratophyllum demersum L., toxicity induced and response of the antioxidant system. Plants were exposed to various concentrations of Cd (0-10 microM) for a period of 1-7 days. Accumulation of Cd was found to be a concentration duration dependent phenomenon. The maximum accumulation of Cd, 1293 microg g(-1) dw, was observed after 7 days at 10 microM. Plants showed significant stimulation of the activities of various antioxidant enzymes viz., superoxide dismutase (EC 1.15.1.1), ascorbate peroxidase (EC 1.11.1.11), guaiacol peroxidase (EC 1.11.1.7), catalase (EC 1.11.1.6), and glutathione reductase (EC 1.6.4.2) and tolerated toxicity of Cd up to moderate concentration of 5 microM. At 10 microM exposure, enzyme activities declined and plants experienced toxicity, which was evident by the significant decrease in the photosynthetic pigments and by increase in the levels of H(2)O(2), lipid peroxidation and ion leakage. In conclusion, modulation of antioxidant system in a coordinated manner in response to Cd accumulation appears to help plants tolerate toxicity of Cd up to 5 microM.

Screening and identification of early warning algal species for metal contamination in fresh water bodies polluted from point and non-point sources.

The water bodies of Lucknow, Unnao and Kanpur (U.P.), India polluted through various point and non point sources were found to be either eutrophic or oligotrophic in nature. These water bodies supported a great number of algal diversity, which varied seasonally depending upon the physico-chemical properties of water. Further, the water bodies polluted through non point sources supports diverse algal species, while the water bodies polluted through point sources supports growth of tolerant blue green algae. High biomass producing algal species growing in these water bodies have accumulated significant amount of metals in their tissues. Maximum amount of Fe was found accumulated by species of Oedogonium sp. II (20,523.00 microg g(-1) dw) and Spirogyra sp. I (4,520.00 microg g(-1) dw), while maximum Chromium (Cr) was found accumulated in Phormedium bohneri (2,109.00 microg g(-1) dw) followed by Oscillatoria nigra (1,957.88 microg g(-1) dw) and Oedogonium sp. I (156.00 microg g(-1) dw) and Ni in Ulothrix sp. (495.00 microg g(-1) dw). Results showed that some of these forms growing in polluted environment and accumulating high amounts of toxic metals may be used as bioindicator species, however, their performance in metal contaminated water under different ecological niche is to be ascertained.