The Effect of Selenite Ions on Growth and Selenium Accumulation in Spirulina platensis

Selenium accumulation and the growth of cyanobacterium Spirulina platensis (Nordst.) Geitl. were studied in a culture with sodium selenite-supplemented nutritional medium. Selenite concentrations below 20 mg/l did not inhibit the growth of S. platensis. The addition of 30 mg/l of this salt somewhat decreased the growth rate during the linear growth phase, induced the earlier suspension transition to the steady-state phase, and substantially lowered the highest optical density of the suspension. However, even at 170 mg/l Na₂SeO₃, the culture still demonstrated a capacity for growth. The content of selenium in the cells depended directly on its concentration in the medium, up to the lethal level. At high selenium concentrations (100–170 mg/l), S. platensis reduced Se(IV) up to Se(0). The latter was secreted onto the cell surface and into the cultural medium. The high concentrations of Na₂SeO₃ acidified the cytoplasmic pH as was measured by ³¹P-NMR spectroscopy. At the same time, the content of protein on a dry weight basis decreased and that of carbohydrates and lipids somewhat increased, just as was observed in S. platensis cells under other stress factors. In the presence of 20 mg/l Na₂SeO₃, the selenium content in the biomass increased by 20000 times as compared to that in the control cells, whereas the biochemical composition of biomass did not change. In this case, the selenium was incorporated almost completely in the protein fraction. The selenium concentration in this fraction increased more significantly when the sulfur content was lowered in the medium.     

Kinetics and mechanism of tolerance induction on acclimation ofVillorita cyprinoides (Hanley) to copper and zinc

The typical euryhaline clamVillorita cyprinoides (Hanley) was acclimated to copper and zinc at salinity 13 × 10⁻³ and < 1 × 10⁻³ (fresh water). Acclimation enhanced the lethal tolerance, as denoted by dose-survival curves, which was more pronounced after zinc acclimation. In fresh water copper acclimation sensitized the organisms. The copper accumulation trend was significantly changed consequent to metal acclimation, especially after zinc acclimation, indicating some tissue metal regulatory effect. Acclimation to copper equiped the organism to survive for longer periods with increased body burden of copper, while zinc acclimation supressed the uptake of the more toxic ion copper. The earlier report of increased uptake of zinc by this organism during combined exposure with copper is corelated in the present context. The role of metallothionein like protein in providing protection against metal toxicity, the environmental implication of acclimation phenomena are indicated     

Biological effects of high copper and zinc concentrations and their interaction in rapeseed plants

In experiments with rapeseed (Brassica napus L., cv. Westar) plants, it was confirmed that copper was considerably more toxic than zinc. The toxic effects of 50 and 150 μM CuSO₄ were comparable to those of 1000 and 2500 μM ZnSO₄. The analysis of the effects of these concentrations of copper and zinc on photosynthetic pigment contents and on the rate of lipid peroxidation did not reveal any reasons for different toxicities of these heavy metals (HM). Among biological effects studied, significant differences were found in the organ distribution of these metals in plants grown on both the standard medium and the medium with high concentrations of copper or zinc. Copper retained in the roots in relatively small amounts and was poorly transported over the aboveground part of the plants. It stayed mainly in the lower leaves, and its distribution changed only a little during the recovery of plants following the HM treatment. In contrast, zinc proved to be highly mobile, it was concentrated in the upper leaves and actively transported when the plants were transferred to a medium with the optimal HM concentrations. High copper concentrations slowed strongly zinc uptake by the roots but practically did not change its movement over the plant. In contrast, high zinc concentrations facilitated copper uptake by the roots but reduced its transfer to the aboveground organs. The data presented here allow us to hypothesize that biological peculiarities of organ and cellular distribution of copper and zinc in plants and interaction of these HM play an important role in the toxic effects of high concentrations of these HM and the mechanisms of adaptation to them at industrial environmental pollution used by rapeseed plants.     

Adaptation of the Common Ice Plant to High Copper and Zinc Concentrations and Their Potential Using for Phytoremediation

A facultative halophite Mesembryanthemum crystallinum L. (the common ice plant) was shown to grow successively at the high concentrations of Cu and Zn. Although 25 μM CuSO₄ or 800 μM ZnSO₄ retarded markedly plant growth, they did not interfere with the completion of plant development and the formation of viable seeds. In such plants, leaves accumulated more than 200 μg of Cu and 1700 μg of Zn per 1 g of dry weight. A damaging effect of heavy metals (HMs) was manifested in a reduced content of water in leaves and proline accumulation in them. As copper is a metal with transient valence, copper salts are more toxic than zinc salts, which was manifested in a stronger inhibition of the chlorophyll synthesis. Both HMs induced oxidative stress, as evident from increased activities of guaiacol peroxidase and lipoxygenase. Moderate Cu and Zn concentrations did not damage cell membranes in leaves, as evident from the absence of their action on electrolyte leakage either under optimum conditions or after heat treatment. A capability of a substantial HM accumulation by the common ice plant and their considerable transport to shoots (up to 50 μg of Cu and 560 μg of Zn per plant) make it possible to consider the common ice plant as a promising phytoremediator. __________ Translated from Fiziologiya Rastenii, Vol. 52, No. 6, 2005, pp. 848–858. Original Russian Text Copyright ? 2005 by Kholodova, Volkov, Kuznetsov.     

Effect of heavy metals combined stress on growth and metals accumulation of three Salix species with different cutting position

This study aimed to compare growth performance and heavy metal (HM) accumulation at different cutting positions of Salix species grown in multi-metal culture. Three Salix species stems cut at different positions (apical to basal) were grown hydroponically for four weeks. The plants were then treated for three weeks with 0, 5, 10, and 20 μM Cd, Cu, Pb, and Zn, resulting in total metal concentrations of 0, 20, 40, and 80 μM. The growth parameters and HM content in shoots and initial cutting were measured. Results showed that, compared with S. fragilis, S. matsudana grew more poorly in uncontaminated condition but grew better and accumulated lower metal in shoots under mixed HM treatment. In addition, cuttings from apical parent stem position exhibited poorer growth performance before and after treatment, as well as greater metal content in shoots than base parts under the HM treatment. These results suggest that S. matsudana may undergo a special mechanism to hinder metals in the initial cutting, thus mitigating growth damage. The apical portion also showed poor resistance against the invasion of mixed HMs because of the immature structure. Therefore, in the selection of phytoremediation plants, metal accumulation ability is not proportional to growth performance.     

Capacity of Spirulina platensis to accumulate manganese and its distribution in cell

Effects of manganese salt (MnCl₂) on growth of Spirulina platensis and capacity of the cyanobacteria to accumulate the metal in various cell components were studied. S. platensis cells were shown to tolerate high concentrations of manganese and preserve, although strongly suppressed, the capacity to grow in the medium containing 5.1 mM MnCl₂. The concentrations of manganese that did not inhibit growth considerably altered cell ultrastructure and changed the protein profile. The accumulation of manganese in S. platensis cells was proportional to the period of culturing and manganese concentration in the medium, reaching a plateau at about 2.5 mM. A threshold intracellular concentration of this metal is estimated as 28 ± 3 μmol/g dry wt. The fractionation of the manganese-enriched biomass demonstrated that the major portion of intracellular manganese (over 90%) was found in the total protein fraction. The chromatographic separation of the soluble protein fraction showed that manganese was incorporated into proteins with molecular weight of 5 to 15 kD. Dry biomass adsorbed manganese cations; this evidence seems to indicate a considerable contribution of biosorption to manganese accumulation by S. platensis cells.     

Vacuolar compartmentalization as indispensable component of heavy metal detoxification in plants

Plant cells orchestrate an array of molecular mechanisms for maintaining plasmatic concentrations of essential heavy metal (HM) ions, for example, iron, zinc and copper, within the optimal functional range. In parallel, concentrations of non‐essential HMs and metalloids, for example, cadmium, mercury and arsenic, should be kept below their toxicity threshold levels. Vacuolar compartmentalization is central to HM homeostasis. It depends on two vacuolar pumps (V‐ATPase and V‐PPase) and a set of tonoplast transporters, which are directly driven by proton motive force, and primary ATP‐dependent pumps. While HM non‐hyperaccumulator plants largely sequester toxic HMs in root vacuoles, HM hyperaccumulators usually sequester them in leaf cell vacuoles following efficient long‐distance translocation. The distinct strategies evolved as a consequence of organ‐specific differences particularly in vacuolar transporters and in addition to distinct features in long‐distance transport. Recent molecular and functional characterization of tonoplast HM transporters has advanced our understanding of their contribution to HM homeostasis, tolerance and hyperaccumulation. Another important part of the dynamic vacuolar sequestration syndrome involves enhanced vacuolation. It involves vesicular trafficking in HM detoxification. The present review provides an updated account of molecular aspects that contribute to the vacuolar compartmentalization of HMs.     

Changes in lipid composition in the tissues of fresh-water plant <Emphasis Type="Italic">Hydrilla verticillata</Emphasis> induced by accumulation and elimination of heavy metals

Qualitative and quantitative composition of lipids was investigated in fresh-water vascular plant Hydrilla verticillata (L. fil.) Royle in the course of the accumulation and elimination of heavy metals (HM). The plants were incubated in 100μM solutions of metal nitrates for 10 days. The accumulation of Cu²⁺, Zn²⁺, and Pb²⁺ and their elimination from the plants depended on the duration of exposure and chemical nature of the metal. Accumulation of lead and copper salts was the greatest on the 3rd day, and zinc, on the 10th day. It was associated with changes in the composition of total lipids, polar lipids, and fatty acid (FA). Copper ions suppressed lipid metabolism stronger than other metals. Zn²⁺ and Pb²⁺ induced the accumulation of biomass and elevated the content of some phospholipids and glycolipids. The detected changes (decrease or increase) were observed both during the incubation with HM and within an afterstress period when the plants recovered in the medium free of metals. Judging by their effect on the content of lipids and FA, HM form a series: Cu²⁺ > Zn²⁺ > Pb²⁺. The responses of plant lipid metabolism to the metals of various chemical nature are discussed.     

Water status in Mesembryanthemum crystallinum under heavy metal stress

Heavy metals (HMs) are known to have negative effects on plant water status; however, the mechanisms by which plants rearrange their water relations to adapt to such conditions are poorly understood. Using the model plant Mesembryanthemum crystallinum, we studied disturbances in water status and rapid plant defence responses induced by excess copper or zinc. After a day of HM stress, reductions in root sap exudation and water deficits in leaf tissues became evident. We also observed several primary adaptive events, including a rapid decrease in the transpiration rate and progressive declines in the leaf-cell sap osmotic potential. Longer HM treatments resulted in reductions of total and relative water contents as well as proline accumulation, an increase in water retention capacity and changes in aquaporin gene expression. After 3 h of HM exposure, leaf expression of the McTIP2;2 gene, which encodes tonoplast aquaporin, was suppressed more than two-fold, thus representing one of the earliest responses to HM treatment. The expression of three additional aquaporin genes was also reduced starting at 9 h; this effect became more prominent upon longer HM exposure. These results indicate that HMs induce critical rearrangements in the water relations of M. crystallinum plants, based on the rapid suppression of transpiration flow and strong inhibition of root sap exudation. These effects then triggered an adaptive water-conserving strategy involving differential regulation of aquaporin gene expression in leaves and roots, further reductions in transpiration, and an accelerated switch to CAM photosynthesis.     

Changes in rhizosphere bacterial populations during phytoextraction of heavy metal contaminated soil with poplar

Abstract

The objective of this paper was to study the response of rhizosphere ammonia‐oxidizing bacterial (AOB) populations during phytoextraction. Hybrid poplars were grown in compartmented root containers with an aged heavy metal (HM)‐contaminated soil for 13 weeks. Bulk and poplar rhizosphere soils were analyzed by denaturing gradient gel electrophoresis (DGGE) of amoA gene fragments. DGGE patterns revealed that amoA‐containing populations in the contaminated soils were markedly different from those in the uncontaminated soils. AmoA profiles appeared to be stable over time in the bulk soils. In contrast, contaminated rhizosphere soils revealed a clear shift of populations with removal of HMs. Rhizosphere AOB populations of the HM‐contaminated soils became similar to the populations of the uncontaminated soils during phytoextraction. The effect of phytoextraction was, however, not evident in the bulk samples, which still contained large amounts of HMs. This study suggests that rhizosphere AOB populations are able to recover after the relief of HM stress by phytoextraction practices.     

Integrative Application of Soil Conditioners and Bio-augmentation for Enhanced Heavy Metal Stabilization from Wastewater and Improved Growth of Nicotiana alata L. and Petunia hydrida L.

Wastewater generated from industries contains numerous contaminants, among which heavy metals (HMs) are non-degradable. This research work highlights the use of commonly used ornamental plants, Nicotiana alata L. and Petunia hydrida L., with compost (C) and peat moss (M), and rhizospheric bacterial augmentation using Pseudomonas japonica, for the phytostabilization of HMs from synthetic wastewater. After plant–soil acclimatization, plants were exposed for 6 weeks to synthetic wastewater, containing cadmium, chromium, copper, lead, nickel, and zinc concentrations (based on the HMs level of wastewaters collected from textile and pharmaceutical industry). Physiological response, biochemical status, and enzymatic fluctuations of plants and the distribution of HMs in plant parts and soil, were quantified. With the combined use (5% each v/v conditioner/soil) of C and M, in bio-augmented soil, physiological response and enzymatic status of both plants improved, with decreased stress injury due to HMs. Further, the plant HMs uptake was reduced, with better stabilization of HM in soil. For better phytostabilization of HMs in wastewater, the use of compost, peat moss, and bacterial augmentation is recommended with Nicotiana alata L. and Petunia hydrida L.

     

Effects of Long-Term exposure to Heavy Metals upon Rhizosphere Bacteria from Baia Mare Area (Maramureş County,Romania)

Abstract

The aim of this study was to investigate the extent of heavy metal (HM) pollution and its effect on microorganisms from rhizosphere soil in Baia Mare area (Maramure? County, Romania). Two sites with different contamination degrees were included in the study: one with a long history of mining activities and one within a drinking water safeguard zone. Rhizosphere soil samples were characterized with respect to physico-chemical parameters and the Cd, Cu, Pb and Zn contents. Native bacteria were investigated for HM tolerance and biofilm formation under toxic exposure by the microdilution assay. The most resistant strains were identified and the minimum inhibitory concentrations for HMs were determined. Cd, Cu, Pb and Zn exceeded the intervention threshold in Bozânta tailings site, while Pb content exceeded the intervention level within the area of the drinking water treatment plant. Cd showed a very high potential ecological risk in Bozânta area. The long-term exposure to HMs contributed to the selection of HM-tolerant and weakly adherent strains. Biofouling was significantly reduced under the influence of copper ions. Arthrobacter, Rhodococcus and Acidovorax strains with exceptional resistant profiles were isolated from the tailings site, indicating the important role of native microorganisms in rhizosphere ecosystems of contaminated sites.     

Mycorrhizal inoculations and silicon fortifications improve rhizobial symbiosis,antioxidant defense,trehalose turnover in pigeon pea genotypes under cadmium and zinc stress

Cadmium (Cd) is a non-essential and highly toxic element for plant growth while zinc (Zn) becomes toxic at elevated levels. Presence of these heavy metals (HMs) in soils has negative impact on rhizobial symbiosis in legumes leading to reduced agricultural productivity. Role of silicon (Si) amendment and Rhizophagus irregularis in mitigating HM stress has gained importance in recent years. Present study evaluated the individual and cumulative effects of Si and/or AM on Cd (25, 50 mg/kg) or Zn (600, 1000 mg/kg) induced responses in terms of nitrogen fixing efficiency, trehalose biosynthesis, antioxidant defense and phytochelatin (PC) synthesis in pigeon pea genotypes (Tolerant-Pusa 2002, Sensitive-Pusa 991). Results indicated that although mycorrhizal colonization (MC) declined with increase in metal concentration in both genotypes, Pusa 2002 was able to form significant colonization even under stress. Cadmium and zinc stress negatively affected plant biomass and rhizobial symbiosis, with Cd more toxic than Zn. The decline in nodulation potential under both HMs was much more significant in Pusa 991 than Pusa 2002 which could be correlated with proportionately reduced MC, nutrient uptake and ultimate N accumulation. Individual application of AM was much more effective in improving nitrogen fixing efficiency by increasing trehalose biosynthesis, PC production and strengthening antioxidant defense than Si. Restoration of rhizobial symbiosis under combined applications of Si and AM could be correlated with enhanced Si uptake through mycorrhization. Thus, study suggested use of AM as a tool in enhancing benefits of Si nutrition in terms of restoration of nodule senescence and N-fixing competence in pigeon pea under HMs stress.     

Phytoextraction of initial cutting of Salix matsudana for Cd and Cu

Salix species are widely used as vegetation filters because of their flourishing root system and fast growth rate. However, studies have yet to determine whether the root system functions in vegetable filters with mixed heavy metal (HM) pollution or whether initial cutting participates in the phytoextraction of HMs. This study aims to determine the function of the root system and initial cutting as vegetation filters in the absorption and accumulation of Cd and Cu. Thick (>1?cm in diameter) and fine (<1?cm in diameter) initial cuttings of Salix matsudana were planted in a nutrient solution with single and mixed (Cd?+?Cu) treatments. The roots of several initial cuttings were removed daily to eradicate rhizofiltration. Results revealed that the existence of the root system altered distribution and interaction of Cd and Cu in plant organs and enhanced tolerance and phytoextraction capacity of plants. The initial cuttings could also absorb and accumulate HMs in the early growth stages of willow without roots. Cu inhibited the plant absorption and accumulation of Cd and promoted Cd transport to shoots. Cd inhibited the Cu absorption of the root system. Our study provided essential data regarding woody species as vegetation filters of HM pollution.     

Cadmium accumulation and interactions with zinc, copper, and manganese, analysed by ICP-MS in a long-term Caco-2 TC7 cell model

The influence of long-term exposure to cadmium (Cd) on essential minerals was investigated using a Caco-2 TC7 cells and a multi-analytical tool: microwave digestion and inductively coupled plasma mass spectrometry. Intracellular levels, effects on cadmium accumulation, distribution, and reference concentration ranges of the following elements were determined: Na, Mg, Ca, Cr, Fe, Mn, Co, Ni, Cu, Zn, Mo, and Cd. Results showed that Caco-2 TC7 cells incubated long-term with cadmium concentrations ranging from 0 to 10 μmol Cd/l for 5 weeks exhibited a significant increase in cadmium accumulation. Furthermore, this accumulation was more marked in cells exposed long-term to cadmium compared with controls, and that this exposure resulted in a significant accumulation of copper and zinc but not of the other elements measured. Interactions of Cd with three elements: zinc, copper, and manganese were particularly studied. Exposed to 30 μmol/l of the element, manganese showed the highest inhibition and copper the lowest on cadmium intracellular accumulation but Zn, Cu, and Mn behave differently in terms of their mutual competition with Cd. Indeed, increasing cadmium in the culture medium resulted in a gradual and significant increase in the accumulation of zinc. There was a significant decrease in manganese from 5 μmol Cd/l exposure, and no variation was observed with copper.     

Effect of light emitting diodes on the cultivation of Spirulina platensis using NPK‐10:26:26 complex fertilizer

NPK‐10:26:26 complex fertilizer based culture medium was studied for the mass production of Spirulina platensis using different light emitting diodes (LEDs). First, cultivation was carried out under white LED to formulate the optimum fertilizer loading for which Spirulina growth was maximized. Optimum composition for newly formulated fertilizer medium was NPK fertilizer ?0.76 g L^?1 and sodium bicarbonate ?10.0 g L^?1 and corresponding biomass productivity was found to be 76.67 mg L^?1 day^?1. The effect of different LEDs (for example, blue, white, red, green and yellow) on Spirulina growth kinetics and the accumulation of chlorophyll, protein and lipid content was determined using the optimum NPK fertilizer medium. Kinetic parameters (i.e., biomass productivity, maximum specific growth rate, maximum biomass concentration, nitrogen‐to‐biomass conversion factor and phosphorus‐to‐biomass conversion factor) and chlorophyll accumulation were affected by the use of different LEDs and follow the following trend: blue > white > red > green > yellow, whereas protein and lipid accumulation was almost independent of LEDs used. Elemental C, N, P and K concentrations were measured to find the effects of nutrients for the growth of Spirulina platensis. Physicochemical parameters (pH and conductivity) were also monitored during biomass growth under different LEDs. Finally, biomass growth using NPK‐10:26:26 fertilizer under different LEDs was compared with standard Zarrouk medium and better growth results were obtained using optimally formulated NPK‐10:26:26 fertilizer medium.     

Physiological and molecular mechanisms of heavy metal accumulation in nonmycorrhizal versus mycorrhizal plants

Uptake, translocation, detoxification, and sequestration of heavy metals (HMs) are key processes in plants to deal with excess amounts of HM. Under natural conditions, plant roots often establish ecto‐ and/or arbuscular‐mycorrhizae with their fungal partners, thereby altering HM accumulation in host plants. This review considers the progress in understanding the physiological and molecular mechanisms involved in HM accumulation in nonmycorrhizal versus mycorrhizal plants. In nonmycorrhizal plants, HM ions in the cells can be detoxified with the aid of several chelators. Furthermore, HMs can be sequestered in cell walls, vacuoles, and the Golgi apparatus of plants. The uptake and translocation of HMs are mediated by members of ZIPs, NRAMPs, and HMAs, and HM detoxification and sequestration are mainly modulated by members of ABCs and MTPs in nonmycorrhizal plants. Mycorrhizal‐induced changes in HM accumulation in plants are mainly due to HM sequestration by fungal partners and improvements in the nutritional and antioxidative status of host plants. Furthermore, mycorrhizal fungi can trigger the differential expression of genes involved in HM accumulation in both partners. Understanding the molecular mechanisms that underlie HM accumulation in mycorrhizal plants is crucial for the utilization of fungi and their host plants to remediate HM‐contaminated soils.