Assessment of chromium phytotoxicity, phytoremediation and tolerance potential of Sesbania sesban and Brachiaria mutica grown on chromite mine overburden dumps and garden soil.

The addition of toxic chromium in agrarian soils from mine overburden dump is of serious concern. To combat the toxicity of chromium an experiment was carried out at the Department of Botany, Utkal University, India by taking the chromium rich overburden dump (OBD) from Sukinda chromite mine, India. Different proportions of chromium rich OBD (10%, 20%, 30%, 40%, 50%, 70% and 100% OBD) were supplied to garden soil and plants of legume and non-legume categories (Sesbania sesban L. and Brachiaria mutica L.) were grown for assessment of phytoaccumulation ability of chromium, tolerance index, chlorophyll, protein and proline and the activity of oxidative inhibitors enzymes. After 60 days of experimentation, Cr phytotoxicity on physiological and biochemical parameters were assessed. The outcome of the results showed that the activity of antioxidant enzymes enhanced considerably in roots as compared to shoots with enhancing concentration of chromium. To compare the phytoaccumulation ability, the tolerance index (TI), bio-concentration factor (BCF) and transportation index (Ti) were measured in two different species. The results showed that at 100% OBD the TI, BCF and Ti of S. sesban have 22.30, 0.45 and 71.06 and B. mutica have 20.83, 0.43 and 42.45 respectively. The result showed that S. sesban not only had the highest bioaccumulation capacity of chromium but also have high tolerance index and transportation index as compared to B. mutica. Hence S. sesban can be recommended for phytostabilization programme to alleviate toxic chromium from chromite overburden mining sites.

Autotoxicity of root exudates varies with species identity and soil phosphorus.

Root exudate autotoxicity (i.e. root exudates from a given plant have toxic effects on itself) has been recognized to be widespread. Here we examined how plant species identity and soil phosphorus (P) availability influenced this autotoxicity and the possible stoichiometric mechanisms. We conducted an experiment with three species (Luctuca sativa, Sesbania cannabina, and Solidago canadensis), which were subject to four treatments consisting of activated carbon (AC) and soil P. AC addition increased the whole-plant biomass of each species under high P conditions and this AC effect varied strongly with species identity. For Solidago, the relative increase in whole-plant biomass due to AC addition was larger in the low P than in the high P. Root exudate autotoxicity differed between roots and shoots. AC addition decreased root N:P ratios but failed to influence shoot N:P ratios in three species. These findings suggest that soil P enrichment might mediate root exudate autotoxicity and that this P-mediated autotoxicity might be related to root N and P stoichiometry. These patterns and their implications need to be addressed in the context of plant communities.

Chromium translocation, concentration and its phytotoxic impacts in in vivo grown seedlings of Sesbania sesban L. Merrill.

The present in vivo pot culture study showed hexavalent chromium (Cr+6) induced phytotoxic impacts and its translocation potential in 21 days old sesban (Sesbania sesban L. Merrill.) seedlings. Cr+6 showed significant growth retardation in 21 days old sesban (Sesbania sesban L. Merrill.) seedlings. Germination of seeds at 10,000 mg L-1 of Cr+6 exhibit 80% inhibition in germination. Seedling survival was 67% after 7 days of seedling exposure to 300 mg kg-1 of Cr+6. Shoot phytotoxicity was enhanced from 6% to 31% with elevated supply of Cr+6 from 10 mg kg-1 to 300 mg kg-1. Elevated supply of Cr+6 exhibited increasing and decreasing trends in % phytotoxicity and seedling tolerance index, respectively. Elevated supply of chromium showed decreased chlorophyll and catalase activities. Peroxidase activities in roots and leaves were significantly higher at increased supply of Cr+6. Cr bioconcentration in roots was nearly 10 times more than stems whereas leaves showed nearly double accumulation than stems. Tissue specific chromium bioaccumulation showed 53 and 12 times more in roots and shoots respectively at 300 mg kg-1 Cr+6 than control. The present study reveals potential of sesban for effective Cr translocation from roots to shoots as evident from their translocation factor and Total Accumulation Rate values.

Effect of lead on phytotoxicity, growth, biochemical alterations and its role on genomic template stability in Sesbania grandiflora: a potential plant for phytoremediation.

The present study was aimed at evaluating phytotoxicity of various concentrations of lead nitrate (0, 100, 200, 400, 600, 800 and 1000mgL(-1)) in Sesbania grandiflora. The seedling growth was significantly affected (46%) at 1000mgL(-1) lead (Pb) treatment. Accumulation of Pb content was high in root (118mgg(-1) dry weight) than in shoot (23mgg(-1) dry weight). The level of photosynthetic pigment contents was gradually increased with increasing concentrations of Pb. Malondialdehyde (MDA) content increased in both the leaves as well as roots at 600mgL(-1) Pb treatment and decreased at higher concentrations. The activity of antioxidative enzymes such as superoxide dismutase and peroxidase were positively correlated with Pb treatment while catalase and ascorbate peroxidase activities increased up to 600mgL(-1) Pb treatment and then slightly decreased at higher concentrations. Isozyme banding pattern revealed the appearance of additional isoforms of superoxide dismutase and peroxidase in Pb treated leaf tissues. Isozyme band intensity was more consistent with the respective changes in antioxidative enzyme activities. Random amplified polymorphic DNA results indicated that genomic template stability (GTS) was significantly affected based on Pb concentrations. The present results suggest that higher concentrations of Pb enhanced the oxidative damage by over production of ROS in S. grandiflora that had potential tolerance mechanism to Pb as evidenced by increased level of photosynthetic pigments, MDA content, and the level of antioxidative enzymes. Retention of high levels of Pb in root indicated that S. grandiflora has potential for phytoextracting heavy metals by rhizofiltration.

Interaction between saflufenacil and imazethapyr in red rice (Oryza ssp.) and hemp sesbania (Sesbania exaltata) as affected by light intensity.

BACKGROUND: Saflufenacil is a broadleaf herbicide for preplant burndown and pre-emergence applications in various crops. This study was established to evaluate the absorption and translocation of saflufenacil in hemp sesbania and imazethapyr in red rice as a function of their post-emergence interaction and light intensity. RESULTS: Imazethapyr plus saflufenacil provided a greater uptake (30%) and translocation (35%) of (14) C-imazethapyr than imazethapyr alone. In the section above treated leaf (ATL), a higher percentage of the absorbed imazethapyr (23%) was quantified in the imazethapyr plus saflufenacil treatment after 168 h. Faster basipetal movement of imazethapyr was identified under higher light availability. Absorption of (14) C-saflufenacil ranged from approximately 40 to 60% among herbicide and light intensity treatments. At 12 and 24 h after treatment (HAT) a greater percentage (15-20%) of the absorbed saflufenacil was quantified above the treated leaf at the two lower light intensities. Similar trends were observed for basipetal movement of saflufenacil. CONCLUSION: Saflufenacil enhanced absorption, overall translocation and acropetal movement of imazethapyr in the TX4 red rice. Basipetal movement of imazethapyr was faster under higher light intensities. Overall, imazethapyr improved absorption of saflufenacil in hemp sesbania plants. Reduction in light intensity resulted in greater translocation of saflufenacil, promoting acropetal and basipetal distribution at the two lower light intensity treatments.

Selected morphological characteristics, lead uptake and phytochelatin synthesis by coffeeweed (Sesbania exaltata Raf.) grown in elevated levels of lead-contaminated soil.

Remediation of lead-contaminated soil is significant due to the inherent toxicity of lead (Pb), and the quantity of Pb discharged into the soil. One of the most cost-effective and environmentally sound technologies for the cleanup of metal-contaminated soils is through the use of plants. While much is known about the ecological evolution of metal tolerance in plants, the physiological, biochemical, and genetic mechanisms of tolerance is not well understood in the majority of resistant ecotypes such as the legume, Sesbania exaltata Raf. This study was therefore conducted to determine the morphological and physiological characteristics of Sesbania that had been grown in Pb-contaminated soil, and to assess phytochelatin synthesis as a way of elucidating its relative Pb tolerance. Sesbania plants were grown in the greenhouse and exposed to various levels of Pb: 0, 1000, and 2000 mg Pb/kg soil. Plants were harvested after 6, 8, and 10 weeks of growth and morphological characteristics (e.g., root and shoot biomass, root length, number of root nodules, shoot height, number of leaves, number of flowers, number and length of pods) were recorded. Generally, there were no statistical differences in morphological characteristics among the treatments. Further, no discernible phytotoxic symptoms, such as chlorosis, wilting, or necrotic lesions, in neither roots nor shoots were observed. We concluded that while Sesbania did not fit the model of a hyperaccumulator, the plant was, nonetheless, tolerant to elevated Pb levels. Our assessment for phytochelatin synthesis as a tolerance mechanism was inconclusive and further investigations of tolerance mechanisms are warranted.

Interactive effects of lead, copper, nickel and zinc on growth, metal uptake and antioxidative metabolism of Sesbania drummondii.

Sesbania drummondii seedlings were grown in a medium to which lead (Pb), copper (Cu), nickel (Ni) and zinc (Zn) were added singly and in combinations in order to assess the effects of metal interactions on seedling growth, metal accumulation and anti-oxidative system. S. drummondii growth was significantly inhibited with metal treatments. S. drummondii accumulated substantially higher concentrations of metals in the roots than shoots. The uptake of metals followed the order Pb>Cu>Zn>Ni in roots and Pb>Zn>Cu>Ni in shoots. In addition, uptake of a single metal by S. drummondii was affected by the presence of a second metal, suggesting an antagonistic effect or competition between metals at the plant uptake site. A significant increase in both enzymatic [superoxide dismutase (SOD), ascorbate peroxidase (APX), glutathione reductase (GR)] and non-enzymatic (glutathione) antioxidant was observed in the S. drummondii seedlings exposed to different metal treatments. The enhancement in enzyme activities followed the order of Cu>Ni>Pb>Zn. However, compared to the effect of individual metal, metals in combination increased the enzyme activities to varying degrees.

Assessing phytotoxicity of heavy metals in remediated soil.

Copper (Cu), zinc (Zn) and chromium (Cr) are pollutants that usually are accumulated in soils. Their toxicity can be decreased by applying amendments. We proposed to evaluate changes in Cu, Zn, and Cr availability, due to the application of amendments, through chemical analysis and phytotoxicity tests. The phytotoxicity test was carried out using species belonging to Sesbania genus; plant parameters were measured 48, 72, 96, and 168 hours after the start of incubation. The treatments included enriched soil, in addition to biosolid compost and triple superphosphate. Cu and Zn amounts were higher in treatments without amendments, indicating immobilization on the part of these. The amounts of Cr tended to decrease with amendments application. The amendments increased pH values and decreased EC; however, this had no impact on the results. No relationship was found among pH, EC, and plant parameters. Different behaviors were observed. S. virgata showed germination seed delay. In addition, while in S. virgata the IG increased during the assay, in S. punicea it diminished. The application of compost, fertilizer or both combined could be of interest for contaminated soils remediation. The use of chemical analysis and phytotoxicity tests allowed to estimate heavy metal availability and the effect on both Sesbania species.

Assessment of the efficacy of chelate-assisted phytoextraction of lead by coffeeweed (Sesbania exaltata Raf.).

Lead (Pb), depending upon the reactant surface, pH, redox potential and other factors can bind tightly to the soil with a retention time of many centuries. Soil-metal interactions by sorption, precipitation and complexation processes, and differences between plant species in metal uptake efficiency, transport, and susceptibility make a general prediction of soil metal bioavailability and risks of plant metal toxicity difficult. Moreover, the tight binding characteristic of Pb to soils and plant materials make a significant portion of Pb unavailable for uptake by plants. This experiment was conducted to determine whether the addition of ethylenediaminetetraacetic acid (EDTA), ethylene glycol tetraacetic acid (EGTA), or acetic acid (HAc) can enhance the phytoextraction of Pb by making the Pb soluble and more bioavailable for uptake by coffeeweed (Sesbania exaltata Raf.). Also we wanted to assess the efficacy of chelates in facilitating translocation of the metal into the above-ground biomass of this plant. To test the effect of chelates on Pb solubility, 2 g of Pb-spiked soil (1000 mg Pb/kg dry soil) were added to each 15 mL centrifuge tube. Chelates (EDTA, EGTA, HAc) in a 1:1 ratio with the metal, or distilled deionized water were then added. Samples were shaken on a platform shaker then centrifuged at the end of several time periods. Supernatants were filtered with a 0.45 mum filter and quantified by inductively coupled plasma-optical emission spectrometry (ICP-OES) to determine soluble Pb concentrations. Results revealed that EDTA was the most effective in bringing Pb into solution, and that maximum solubility was reached 6 days after chelate amendment. Additionally, a greenhouse experiment was conducted by planting Sesbania seeds in plastic tubes containing top soil and peat (2:1, v:v) spiked with various levels (0, 1000, 2000 mg Pb/kg dry soil) of lead nitrate. At six weeks after emergence, aqueous solutions of EDTA and/or HAc (in a 1:1 ratio with the metal) or distilled deionized water were applied to the root zones. Plants were harvested at 6 days after chelate addition to coincide with the duration of maximum metal solubility previously determined in this study. Results of the greenhouse experiment showed that coffeeweed was relatively tolerant to moderate levels of Pb and chelates as shown by very slight reductions in root and no discernable effects on shoot biomass. Root Pb concentrations increased with increasing levels of soil-applied Pb. Further increases in root Pb concentrations were attributed to chelate amendments. In the absence of chelates, translocation of Pb from roots to shoots was minimal. However, translocation dramatically increased in treatments with EDTA alone or in combination with HAc. Overall, the results of this study indicated that depending on the nature and type of Pb-contaminated soil being remediated, the bioavailability and uptake of Pb by coffeeweed can be enhanced by amending the soil with chelates especially after the plants have reached maximum biomass.

Bioavailability and uptake of lead by coffeeweed (Sesbania exaltata Raf.).

Lead (Pb) is recognized as one of the most pervasive environmental health concerns in the industrialized world. While there has been a substantial reduction in the use of Pb in gasoline, water pipes, and Pb-based residential paint, residual Pb from their use is still in the environment and constitutes an important source of Pb in the atmosphere, water, and soil. Soil acts as a sink for these anthropogenic sources of Pb, accumulating the deposits over time in the upper 2 - 5 cm of undisturbed soil. Generally, Pb binds strongly to soil particles and renders a significant soil-metal fraction insoluble and largely unavailable for phytoremediation or plant uptake. A major objective of current phytoremediation research, therefore, is to induce desorption of Pb from the soil matrix into solution and increase the propensity for plant uptake. We hypothesized that the bioavailability of Pb for plant uptake can be increased through chelate amendments. To test this hypothesis, we mixed delta top soil and peat (2:1) and added lead nitrate [Pb (NO3)2[ to generate a Pb-contaminated soil concentration of 2000 mg Pb/kg dry soil. After incubating the Pb-spiked soil in a greenhouse for 6 weeks, Sesbania plants were grown in the soil and harvested at 6, 8, and 10 weeks after emergence. Six days before each harvest, a chelating agent, ethylenediaminetetraacetic acid (EDTA) was applied to the root zone as an aqueous solution in a 1:1 ratio with the Pb concentration in the soil. Sequential extraction procedures were used to assess selective chemical fractions of Pb in the soil. Our results showed that a higher exchangeable fraction of Pb was available for plant uptake after chelate amendment compared to pre-chelate amendment. We also saw higher root and shoot Pb uptake after chelate amendment compared to pre-chelate amendment, especially at 10 weeks after emergence. Together, these results suggest that chelate amendments can promote the bioavailability of Pb in the soil and increased the propensity for uptake by plants into roots and shoots. Further, these results indicate that Sesbania exaltata can be grown under elevated Pb conditions and may be suitable as a potential crop rotation species for phytoextraction.

Identification of lead-regulated genes by suppression subtractive hybridization in the heavy metal accumulator Sesbania drummondii.

Heavy metal contamination of soils is of widespread occurrence as a result of human, agricultural and industrial activities. Among heavy metals, lead is a potential pollutant that readily accumulates in soils and sediments. Although lead is not an essential element for plants, it gets easily absorbed and accumulated in Sesbania drummondii, which exhibits a significant level of tolerance to lead. The response of a metal tolerant plant to heavy metal stress involves a number of biochemical and physiological pathways. To investigate the overall molecular response of a metal-tolerant plant to lead exposure, suppression subtractive hybridization (SSH) was used to construct a cDNA library enriched in lead induced mRNA transcripts from lead-tolerant Sesbania. Screening the library by reverse Northern analysis revealed that between 20 and 25% of clones selected from the library were differentially regulated in lead treated plants. After differential screening, we isolated several differentially expressed cDNA clones, including a type 2 metallothionein (MT) gene which is involved in detoxification and homeostasis and shown to be differentially regulated in lead treated plants. The data from the reverse Northern analysis was further confirmed with conventional Northern analysis of a select group of genes including MT, ACC synthase/oxidase, cold-, water stress-, and other abiotic stress-induced genes, which are up-regulated rapidly in response to lead treatment. The mRNA levels of MT increased substantially after lead treatment indicating a potential role for it under lead stress in Sesbania. The present results show that SSH can serve as an effective tool for isolating genes induced in response to lead heavy metal tolerance in Sesbania. A better understanding of lead induced gene expression in Sesbania should help select candidates associated with remediation of heavy metal toxicity. The possible link between this result and the heavy-metal response of plants is discussed.

Antioxidative responses to mercury in the cell cultures of Sesbania drummondii.

The effect of mercury (Hg) on the growth and the response of antioxidative systems have been investigated in Sesbania cell cultures to determine the tolerance limits and the mechanisms of metal (Hg) tolerance in plant cells. Cell cultures of Sesbania were developed in different concentrations (0-50 microM) of mercury. Cultures tolerated Hg up to a concentration of 40 microM and showed an increase in the fresh weight growth by 620% in 3 weeks. The levels of antioxidants: glutathione (GSH) and non-protein thiols (NPSH) and the activities of antioxidative enzymes: superoxide dismutase (SOD, EC 1.15.1.1), ascorbate peroxidase (APX, EC 1.11.1.11) and glutathione reductase (GR, EC 1.6.4.2) were influenced by Hg treatments. The contents of GSH, NPSH and GSH/GSSG ratio increased up to a concentration of 40 muM Hg and then severely declined at 50 microM Hg. The activities of antioxidative enzymes, SOD, APX and GR followed the same trends as antioxidants, first increased up to a concentration of 40 muM Hg and then declined in the presence of 50 microM Hg.

Effects of lead and chelators on growth, photosynthetic activity and Pb uptake in Sesbania drummondii grown in soil.

Effects of lead (Pb) and chelators, such as EDTA, HEDTA, DTPA, NTA and citric acid, were studied to evaluate the growth potential of Sesbania drummondii in soils contaminated with high concentrations of Pb. S. drummondii seedlings were grown in soil containing 7.5 g Pb(NO(3))(2) and 0-10 mmol chelators/kg soil for a period of 2 and 4 weeks and assessed for growth profile (length of root and shoot), chlorophyll a fluorescence kinetics (F(v)/F(m) and F(v)/F(o)) and Pb accumulations in root and shoot. Growth of plants in the presence of Pb+chelators was significantly higher (P<0.05) than the controls grown in the presence of Pb alone. F(v)/F(m) and F(v)/F(o) values of treated seedlings remained unaffected, indicating normal photosynthetic efficiency and strength of plants in the presence of chelators. On application of chelators, while root uptake of Pb increased four-five folds, shoot accumulations increased up to 40-folds as compared to controls (Pb only) depending on the type of chelator used. Shoot accumulations of Pb varied from 0.1 to 0.42% (dry weight) depending on the concentration of chelators used.

Cadmium accumulation and antioxidative responses in the Sesbania drummondii callus.

The effect of cadmium (Cd) on growth, accumulation, and antioxidative response was studied in Sesbania drummondii callus, cultivated on different concentrations of Cd (0-250 microM) for four weeks. Callus growth was comparable to that of the control for concentrations up to 50 microM Cd; however, concentrations higher than 50 microM affected growth. A concentration of 100 microM Cd inhibited growth by 16%, with respect to control. Cd concentration in callus increased with increasing Cd concentrations in the growth medium. Callus accumulated 530 mg Cd kg(-1) of their dry weight at 100 microM Cd concentration. Sesbania callus responded to Cd-induced oxidative stress by modulating antioxidants (glutathione and other non-protein thiols) level and antioxidative enzymes: superoxide dismutase (SOD), ascorbate peroxidase (APX), and glutathione reductase (GR). The content of the glutathione (GSH) and GSH/GSSG ratio first increased up to a concentration of 50 microM Cd and then decreased. The content of other non-protein thiols significantly increased with increasing Cd concentrations in the growth medium. The activities of antioxidative enzymes, SOD, APX, and GR, followed the same trends as antioxidants first increasing up to a concentration of 50 microM Cd and then decreasing. These results suggest that antioxidative defense mechanisms play a significant role in Cd detoxification and accumulation in Sesbania drummondii.

Translocation of metals from fly ash amended soil in the plant of Sesbania cannabina L. Ritz: effect on antioxidants.

The plants of Sesbania cannabina Ritz grown on different amendments of fly ash (FA), have shown a high accumulation of metals (Fe, Mn, Zn, Cu, Pb and Ni). The highest accumulation of Fe the and lowest level of Ni were recorded in these plants. The different amendments of fly ash with garden soil (GS) were extracted with DTPA and the levels of metals were found to be decreased with an increase in fly ash application ratio from 10% to 50% FA. The analysis of the results showed an increase in the level of malondialdehyde (MDA) content of the roots for all the exposure periods. The maximum increases of 136% (roots) and 120% (leaves) were observed in MDA content at 100% FA after 30 d of growth of the plant, compared to GS. The level of antioxidants was found to increase for all the exposure periods in the roots of the plants to combat metal stress. At 30 d, the maximum increase of 57% (ascorbic acid) and 78% (free proline) was observed in the roots of the plants grown on 100% and 10% FA, respectively, as compared to their respective GS. At 90 d, a maximum increase of 42% (cysteine) and 117% (NPSH) was recorded in the roots of the plants grown on 25% and 100% FA, respectively, as compared to their respective GS. In leaves, a significant increase in antioxidants i.e. cysteine, NPSH and free proline content was recorded after 30 d, whereas no such trend was observed for the rest of the exposure periods. The chlorophyll and carotenoid contents increased with an increase in the FA amendment ratio from 10% to 50% FA for all the exposure periods as compared to GS. In both roots and leaves, the level of protein content increased in all the amendments and 100% FA at 30 d as compared to GS. Thus, there is a balance in the level of MDA content and level of antioxidants in the plants at 90 d. In view of its tolerance, the plants may be used for phytoremediation of metals from fly ash contaminated sites and suitable species for plantation on fly ash land fills.