Integrating Nonindigenous Aquatic Plant Control with Protection of Snail Kite Nests in Florida

The endangered snail kite (Rostrhamus sociabilis) feeds primarily on the freshwater apple snail (Pomacea paludosa) in Florida. The nonindigenous, floating water hyacinth (Eichhornia crassipes) and water lettuce (Pistia stratiotes) impede kites from finding snails. Effective control of these aquatic plants in the littoral zone of central and south Florida lakes benefits kites by maintaining open foraging habitat. However, incidental herbicide spraying of nesting substrates result in nest collapse when kites breed in nonwoody, emergent plants [cattail (Typha spp.) and giant bulrush (Scirpus validus)] in the outer littoral zone during lower lake levels. Many endangered species recovery plans and their implementation have experienced problems due to inaction and/or noncooperation by various governmental agencies and their personnel. Herein, we describe the development and implementation of a buffer zone strategy to prevent secondary impacts from an aquatic plant control program to snail kites nesting on lakes in central and south Florida. A strategy was jointly developed by personnel of five state and federal agencies to control herbicide application near kite nesting areas during the normal breeding season. Although requiring various modifications during its implementation, this cooperative effort successfully integrated aquatic plant control objectives with snail kite conservation on Lake Okeechobee during 1988. The program was expanded the following year to lakes Kissimmee and Tohopekaliga. Since the implementation of the snail kite impact preclusion program, no nest loss was attributed to incidental herbicide applications on lakes Okeechobee, Kissimmee, and Tohopekaliga.     

Does radioactive contamination affect the shell morphology of the pond snail <Emphasis Type="Italic">Lymnaea stagnalis</Emphasis> in the exclusion zone of the Chernobyl NPP (Ukraine)?

A distinct external shell abnormality, coarse seam, was found in a high percentage of the pond snail Lymnaea stagnalis (L.) collected near Chernobyl Nuclear Power Plant (NPP), Ukraine. The negative cytogenetical and hematological effects of long-term irradiation on pond snails in the Chernobyl Exclusion Zone have been repeatedly reported; consequently, it has been assumed that radioactivity may also affect shell morphology. However, due to the absence of this shell abnormality in pond snails collected from two radioactive sites in Russia, it can be concluded that the appearance of abnormal shells in snail population near Chernobyl NPP cannot directly be explained by the radioactive contamination.     

Glutathione Related Enzyme Levels of Freshwater Fish as Bioindicators of Pollution

The livers and kidneys of freshwater fish species, Oreochromis niloticus and Clarias lazera, collected from sewage polluted sites (Ismailia and El-Bahr El-Azam) and industrial polluted sites (Shubra and El-Tebin) of Nile River were analyzed for different antioxidant defense enzymes. The liver and kidney glutathione transferase (GST), glutathione reductase (GR) and glutathione peroxidase (GPx) were higher in O. niloticus captured from all the polluted areas compared to the control. Low GST activities were found in 33.3%, 60% and 53% in the livers and 100%, 80% and 53% in the kidneys of C. lazera captured from El-Bahr El-Azam, Shubra and El-Tebin. GR and GPx activities increased in livers and kidneys of C. lazera collected from all areas except for Shubra, in which, GPx of livers and kidneys were low in 100% of C. lazera. Metals Ni, Co, Cr, Se, Cd and Pb resulting from industrial wastes and metal mining wastes were enhanced at the polluted sites. SDS-PAGE of liver and kidney of O. niloticus and C. lazera indicated the increase in bands number and intensity of protein bands with subunit molecular weights between 30–20 KDa in polluted areas. Several enzymes from glutathione system (activity and protein) constitute a sensitive biochemical indicator of chemical pollution. Relative changes of glutathione-dependent enzymes in both fish species suggest a different susceptibility to toxins.     

Evaluation of tidal marsh restoration: Comparison of selected macroinvertebrate populations on a restored impounded valley marsh and an unimpounded valley marsh within the same salt marsh system in Connecticut,USA

Macroinvertebrates were examined on an impounded valley marsh in Stonington, Connecticut, that has changed from aTypha-dominated system to one with typical salt-marsh vegetation during 13 years following the reintroduction of tidal exchange. Animal populations on this restored impounded marsh were evaluated by comparing them with populations on a nearby unimpounded valley marsh of roughly the same size. Populations of the high marsh snail,Melampus bidentatus Say, were quantitatively sampled along transects that extended from the water-marsh edge to the upland; those of the ribbed mussel,Geukensia demissa Dillwyn, were sampled in low marsh areas on transects along the banks of creeks and mosquito ditches. The occurrence of other marsh invertebrates also was documented, but their abundance was not measured. The mean density ofMelampus was 332±39.6 SE/m² on the restored impounded marsh and 712±56.0 SE/m² on the unimpounded marsh. However, since snails were larger on the restored impounded marsh, the difference in snail biomass was less pronounced than the difference in snail density. MeanMelampus biomass was 4.96±0.52 SE g dry wt/m² on the restored impounded marsh and 6.96±0.52 SE g dry wt/m² on the unimpounded marsh. On the two marshes, snail density and biomass varied in relation to plant cover and other factors. The density and biomass ofGeukensia at the edge of the marsh were comparable on the restored impounded and unimpounded marshes. Mean mussel densities ranged from 80 to 240/m² and mean mussel biomass varied from 24.8–64.8 g dry wt/m² in different low marsh areas. In contrast, below the impoundment dike, meanGeukensia density was 1100±96.4 SE/m² and meanGeukensia biomass was 303.6±33.28 SE g dry wt/m². A consideration of all available evidence leads to the conclusion that the impounded marsh is in an advanced phase of restoration.     

Application of magnetic field and iron in order to change medicinal products of <Emphasis Type="Italic">Ocimum basilicum</Emphasis>

In the present research, the effects of static magnetic field, with or without iron on the growth, the activity of certain enzymes (like polyphenol oxidase and phenyle alanine ammonialyase), the content of phenolic compounds and the essential oil composition of sweet basil (Ocimum basilicum) were studied. Treatment of plants with static magnetic field with or without iron led to decrease of the growth of plants as well as decrease in the activity of phenyle alanine ammonialyase and of phenolic compounds content. In addition in those plants exposed to magnetic field the amount of essential oils, notably of methyl chavicol, was increased. Exposure of plants to the magnetic field decreased the activity of polyphenol oxidase, whereas treatment of plants with iron when exposed to magnetic field increased polyphenol oxidase activity. In this group the content of essential oil was reduced.     

Resistance of pulmonate snail populations to repeated treatments of copper sulfate

Two species of pulmonate snails,Lymnaea cat-ascopium andPhysa integra, were collected from Douglas and Houghton Lakes. Snail populations from the former lake (pristine) had never been exposed to copper sulfate, whereas those from the latter one (treated) had been subjected to the molluscicide for more than 40 years. Molluscs from the treated lake were more resistant to the copper at concentrations of 0.5 and 1.0 ppm than those from the pristine lake. Furthermore, larger lymnaeids had a higher survival rate than smaller ones.     

AQUATIC ORGANISMS RESPONSE TO SEVERE STRESS FOLLOWING ACUTELY SUBLETHAL TOXICANT EXPOSURE1

ABSTRACT: Snails, Goniobasis livescens (Menke), were exposed to acutely sublethal concentrations of p-nitrophenol and the lampricide, 3-triflourmethyl-4 nitrophenol(TFM), and then to acutely lethal thermal shocks. The same species were also exposed to acutely sublethal concentrations of zinc followed by exposure to acutely lethal concentrations of p-nitrophenol; and to acutely sublethal concentrations of TFM followed by exposure to acutely lethal concentrations of zinc. Brown trout, Salmo trutta, were exposed to acutely sublethal concentrations of TFM and then to an acutely lethal thermal shock. Results of these experiments indicate that prior exposure to acutely sublethal toxicant concentrations may reduce survival time for a subsequent exposure to acutely lethal concentrations of a second toxicant (e.g., snails exposed to Zn⁺⁺ then p-nitrophenol) but neither prior or concomitant exposure to acutely sublethal toxicant concentrations insures that the median survival time for a lethal exposure will be significantly altered (e.g., snails exposed to 0.2 × 48 hour LC50 for TFM then Zn⁺⁺). However, some acutely sublethal concentrations of a toxicant may significantly alter survival time of snails to a lethal concentration of a different toxicant (e.g., exposure to 0.4 × 48 hour LC50 TFM then a lethal dose of Zn⁺⁺). The brown trout exposed to an acutely sublethal concentration of TFM and then an acutely lethal thermal shock did have significantly altered survival patterns.     

ACID MINE DRAINAGE (AMD) AND ITS IMPACT ON A SMALL VIRGINIA STREAM1

ABSTRACT: A study was conducted to 1) chemically characterize three AMD streams entering Chestnut Creek in southwestern Virginia below the town of Galax, Virginia, and 2) to assess the biological and chemical alterations in the creek. Over a six mile reach, the benthic macroinvertebrate population was reduced to zero, the naturally low alkalinity (～25mg/1) of the stream was reduced to less than 5 mg/1, and the pH was reduced from 7.2 to 6.3. Increased concentration of iron from less than 0.01 mg/I to more than 4.0 mg/1 were accompanied by the deposition of a coating of iron hydroxide up to 0.25 in. thick in the stream bed, a phenomenon most likely responsible for the absence of benethic macroinvertebrates. In situ bioassays with bluegill sunfish and one snail species showed that the creek water, after confluence with all the AMD streams, was not toxic in 192 hours to fish, and snails survived 96 hours before they began to die. The undiluted AMD itself was highly toxic.     

Agave salmiana Plant Communities in Central Mexico as Affected by Commercial Use

Agave salmiana is a native plant species harvested for the commercial production of mezcal (Agave spirits) in the highlands of central Mexico. The objective of this study was to identify vegetation changes in natural communities where A. salmiana has been differentially harvested for commercial purposes. Three plant community categories were identified in the state of Zacatecas based on their history of A. salmiana utilization: short (less than 10 years of use), moderate (about 25 years), and long (60 or more years). Species cover, composition, and density were evaluated in field surveys by use category. A gradient of vegetation structure of the communities parallels the duration of A. salmiana use. A. salmiana density was greatest (3,125 plants ha⁻¹) in the short-use areas and less (892 plants ha⁻¹) in the moderate-use areas, associated with markedly greater density of shrubs (200%) and Opuntia spp. (50%) in moderate-use areas. The main shrubs were Larrea tridentata, Mimosa biuncifera, Jatropha dioica and Buddleia scordioides while the main Opuntia species were Opuntia leucotricha and Opuntia robusta. A. salmiana density was least (652 plants ha⁻¹) in the long-use areas where shrubs were less abundant but Opuntia spp. density was 25% higher than in moderate-use areas. We suggest that shrubs may increase with moderate use creating an intermediate successional stage that facilitates the establishment of Opuntia spp. Long-term Agave use is generating new plant communities dominated by Opuntia spp. (nopaleras) as a replacement of the original communities dominated by A. salmiana (magueyeras).     

Biochemical responses,growth and reproduction of earthworm in low density polyethylene (LDPE)

There is an unprecedented production of plastic that is accelerating its disposal while affecting the fitness of the terrestrial as well as the aquatic environment. The term microplastics refers to plastic fragments that are less than 5 mm in size and are widely distributed in the environment. Therefore, the present study intends to explore the biological response of earthworms (Eisenia fetida) toward different concentrations of low-density polyethylene. E. fetida treated with low-density polyethylene concentration (Control), 250 mg kg⁻¹, 1000 mg kg⁻¹, 6000 mg kg⁻¹, 12,000 mg kg^−1, and 25,000 mg kg⁻¹. The above ratios were thoroughly mixed with 1kg of artificial soil and tested for growth, reproduction (cocoons and hatchling count), and enzymatic activities namely superoxide dismutase, guaiacol peroxidase, glutathione-S-transferase, and glutathione reductase and molecular docking studies. No mortality was observed during the exposure period at any concentrations. On the 28th day, when compared to the control the highest decrease in body weight of earthworms was observed in 25,000 mg (28.4%) followed by 12,000 mg (12.2%) and 6000 mg (3.4%). The cocoon and hatchlings significantly declined as the dose of microplastics increases. Enzymatic activity such as SOD and POD showed declined trend as the dose increased, while GST and GR increased with an increase in microplastic concentrations on 28th day. Furthermore, molecular docking showed that LDPE can modulate the activity of all four enzymes significantly.     

Macroinvertebrate and fish populations in a restored impounded salt marsh 21 years after the reestablishment of tidal flooding

During the last two decades, the State of Connecticut has restored tidal flow to many impounded salt marshes. One of the first of these and the one most extensively studied is Impoundment One in the Barn Island Wildlife Management Area in Stonington, Connecticut. In 1990, twelve years after the re-establishment of tidal flooding, the density of the marsh snail Melampus bidentatus, the numerically dominant macroinvertebrate of the high marsh, in Impoundment One was about half that in reference marshes below the breached impoundment dike. By 1999 the densities of Melampus above and below the dike were not significantly different, but the shell-free biomass was greater above the dike as a result of the somewhat larger number and size of the snails there. Twenty-one years after the renewal of tidal flooding, three marsh macroinvertebrates (the amphipods Orchestia grillus and Uhlorchestia spartinophila and the mussel Geukensia demissa) were significantly less abundant in the previously impounded marsh than in the reference marshes, whereas another amphipod (Gammarus palustris) was more abundant above the breached dike where conditions appeared to be somewhat wetter. In 1991 the fish assemblage in a mosquito-control ditch in Impoundment One was similar to that in a ditch below the breached dike; however, the common mummichog Fundulus heteroclitus appeared to be less abundant in the restoring marsh. By 1999 the number of mummichogs caught in ditches was significantly greater in Impoundment One than in the reference marsh, but the numbers of mummichogs trapped along the tidal creek were comparable above and below the dike. The results obtained in this study and those of other restoring marshes at Barn Island indicate the full recovery of certain animal populations following the reintroduction of tidal flow to impounded marshes may require up to two or more decades. Furthermore, not only do different species recover at different rates on a single marsh, but the time required for the recovery of a particular species may vary widely from marsh to marsh, often independently of other species.     

THE USE OF OVERWINTER DRAW DOWN FOR AQUATIC VEGETATION MANAGEMENT1

ABSTRACT: Overwinter draw down can be a useful technique for aquatic plant management. Its effectiveness depends largely on the susceptibility of nuisance species to draw down. A single overwinter draw down provided good control of aquatic plants in a flowage dominated by Potamogeton robbinsii Little additional control was gained by a second draw down the following winter. Rapid reinvasion of plants after draw down ceased dictates continued management. To avoid fish kills caused by low dissolved oxygen levels caution is advised when using overwinter draw down. The growth of Zizania aquatica was not negatively influenced by draw down. The influence on water quality of nutrient release from decaying vegetation and exposed bottom sediments was uncertain.     

Protective influence of Vitamins against Petroleum-induced free radical toxicity in Rabit

The effect of exposure to crude oil contaminated diet on the blood antioxidant defence system, lipid peroxidation, lipid profile as well as possible protective roles of vitamins E and C were studied in rabbits. Oxidative stress induction by crude oil was indicated by significantly (P < 0.05) increased lipid peroxidation and a non-significant (P < 0.05) decrease in superoxide dismutase and catalase activities. A similar pattern was also detected in the lipid profile: total cholesterol and LDL-cholesterol insignificantly (P < 0.05) increased while HDL-cholesterol and triglyceride significantly decreased relative to rabbits fed normal diet. The reciprocal relationship between HDL-cholesterol and LDL-cholesterol in addition to compromised antioxidant enzymes could predispose exposed animals to coronary heart disease. However, pre-treatment of the diet with vitamins C and E exhibited a protective role on the toxic effect of crude oil on lipid profile, lipid peroxidation as well as antioxidant enzymes. The order of protection was vitamin E + C > vitamin E > vitamin C. These observations seemed to suggest that the protective role of vitamins C and E is synergistic. The protective role of the vitamins is probably time-dependent as significant (P < 0.05) restoration of lipid profile as well as antioxidant enzymes activities to control values was effected after four weeks of exposure. It is therefore suggested that toxic effect of crude oil may be reduced by dietary supplementation of vitamins C and E.     

Genotypic Variation in the Phytoremediation Potential of Indian Mustard for Chromium

The term “phytoremediation” is used to describe the cleanup of heavy metals from contaminated sites by plants. This study demonstrates phytoremediation potential of Indian mustard (Brasicca juncea (L.) Czern. & Coss.) genotypes for chromium (Cr). Seedlings of 10 genotypes were grown hydroponically in artificially contaminated water over a range of environmentally relevant concentrations of Cr (VI), and the responses of genotypes in the presence of Cr, with reference to Cr accumulation, its phytotoxity and anti-oxidative system were investigated. The Cr accumulation potential varied largely among Indian mustard genotypes. At 100 μM Cr treatment, Pusa Jai Kisan accumulated the maximum amount of Cr (1680 μg Cr g⁻¹ DW) whereas Vardhan accumulated the minimum (107 μg Cr g⁻¹ DW). As the tolerance of metals is a key plant characteristic required for phytoremediation purpose, effects of various levels of Cr on biomass were evaluated as the gross effect. The extent of oxidative stress caused by Cr stress was measured as rate of lipid peroxidation. The level of thiobarbituric acid reactive substances (TBARS) was enhanced at all Cr treatments when compared to the control. Inductions of enzymatic and nonenzymatic antioxidants were monitored as metal-detoxifying responses. All the genotypes responded to Cr-induced oxidative stress by modulating nonenzymatic antioxidants [glutathione (GSH) and ascorbate (Asc)] and enzymatic antioxidants [superoxide dismutase (SOD), ascorbate peroxidase (APX), and glutathione reductase (GR)]. The level of induction, however, differed among the genotypes, being at its maximum in Pusa Jai Kisan and its minimum in Vardhan. Pusa Jai Kisan was grown under natural field conditions with various Cr treatments, and Cr-accumulation capacity was studied. The results confirmed that Pusa Jai Kisan is a hyperaccumulator of Cr and hypertolerant to Cr-induced stress, which makes this genotype a viable candidate for use in the development of phytoremediation technology of Cr-contaminated sites.     

Chromosomal genes conferring tolerance to heavy metal (Ag) toxicity

Bacterial strains were isolated from sediment samples from the Thames River. Successive transfer growth of the various strains on nutrient agar containing increasing concentrations of AgNO₃ revealed that three of the bacterial isolates were found to be capable of tolerating high concentrations of AgNO₃ ranging from 20 to 80 mM on a solid medium and up to 10 mM AgNO₃ in liquid medium. Molecular characterization and identification based on 16S rDNA gene sequencing of three strains of bacteria that are tolerant to silver nitrate showed that the major tolerant strains include the superbug, Shewanella oneidensis, Pseudomonas sp. and Bacillus sp. Protein extraction and two-dimensional (2D) sodium dodecyl sulfate SDS-polyacrylamide gel electrophoresis (PAGE) of the protein extracts in bacteria exposed to very high concentrations of AgNO₃ revealed a general reduction in the number of expressed proteins, although two protein spots were conspicuously over expressed in the exposed bacteria compared to control. The N-terminal amino acid sequence analysis of the protein spots identified the major up-regulated proteins as the outer membrane protein To1C (45.2 kDa) and the structural protein of the flagellar filament, flagellin (28.34 kDa), encoded for by the to1C and fliC genes, respectively. The roles of these genes in a number of multi-drug resistant pathogen and potentials for biotechnological applications in toxic metal control for treatment of contaminated ecosystems and biomining were discussed.     

Radical Scavengers Suppress Low Frequency EMF Enhanced Proliferation in Cultured Cells and Stress Effects in Higher Plants

Summary In previous publications, we reported that sinusoidal varying magnetic fields (SVMF) modify the activity and dynamics of the malignancy marker adenosine deaminase, and enhance the proliferation of chick embryo fibroblasts (CEF). While the SVMF examined by us (50, 60 & 100 Hz / 0.06–0.7 mT) were all below kT, they may have the potential of altering chemical processes in which excited radicals are involved. We tested this hypothesis in two experimental systems: CEF in culture and Spirodela oligorrhiza (Lemnaceae) (a small aquatic plant, commonly known as Duckweed). CEF were exposed to SVMF of 100 Hz/0.7 mT for 24 h. The addition of the exogenous radical scavengers catalase, superoxide dismutase or vitamin E to the cells during exposure significantly suppressed enhancement of cell proliferation caused by the field (by 79, 67 and 82%, respectively, as evaluated by the MTT colorimetric assay). ¹⁵N NMR analysis of Duckweed plants fed by ¹⁵N-labeled ammonium chloride and exposed to SVMF at 60 and 100 Hz/0.7 mT for 24 h, revealed augmented alanine production. Alanine did not accumulate in the absence of SVMF. The addition of vitamin C, a radical scavenger, reduced alanine production by 82%. Exposure to SVMF resulted in specific metabolic stress effects in Duckweed plants and enhanced proliferation of CEF. In both cases, it is suggested that free radicals are involved.     

EVALUATION OF COPPER ACCUMULATION IN PART OF THE CALIFORNIA AQUEDUCT1

ABSTRACT: Copper sulfate has been used extensively in the California Aqueduct to control phytoplankton and the alga Cladophora. Since 1969 more than 250,000 pounds of copper sulfate has been added to a part of the aqueduct. Although copper sulfate is effective in controlling algae, copper tends to accumulate in the system in which it is applied. Samples of water, biota including plants, clams, and snails, as well as sediment were analyzed for copper. Results of these analyses showed that copper concentrations in the water, with three exceptions, were less than 10 micrograms per litre. Samples of plant tissue showed a 198 percent increase in copper concentration in the treated reach of the aqueduct, and clam tissue showed a 68 percent increase. Snails in the treated reach had 77 percent more copper than in the untreated reach whereas copper concentrations in the sediment doubled. None of the concentrations found are considered to be harmful to the biota.     

Physiological responses of Avicennia marina seedlings to the phytotoxic effects of the water-soluble fraction of light Arabian crude oil

Stomatal behavior, growth performance and the accumulation of polynuclear aromatic hydrocarbons (PAHs) were evaluated in seedlings of the mangrove Avicennia marina (Forssk.) Vierh., treated with a water-soluble fraction (WSF) of Abu-Dhabi light Arabian crude oil through foliar spraying or soil application.Irregular stomatal behavior and weak stomatal control over transpiration were observed during the first 24 hours, where stomatal resistances of plants sprayed with 150 and 300 g PAHs plant^–1 were significantly lower than that of the control plants. After six weeks, all treated plants showed no significant difference in their relative growth rate (RGR) or in the net assimilation rate (NAR) compared with the control plants.Tri-aromatic hydrocarbons were the most accumulated in tissues of the treated plants. Penta- and hexa-aromatics, on the other hand, were undetectable in the WSF and consequently in the treated plants. A linear relationship was observed between the dose applied to plants and the amounts of tissue accumulated PAHs (r ²=0.515 for soil application and r ²=0.984 for foliar spray). In plants sprayed with 300 g PAHs plant^–1, the total PAHs accumulated were more than that accumulated in plants treated through soil application.These findings suggest that: aqueous extraction of crude oil tends to signify the percentage of the low molecular weight PAHs, e.g. naphthalene, to the total PAHs; disturbed stomatal behavior in the first day of the treatment may be due to the venting of the volatile low molecular weight aromatic hydrocarbons (e.g. benzene, toluene, and xylenes) through the stomata; and uptake of water-soluble hydrocarbons by plants is equally possible through both of the root system and the foliage. The ecological implications of these finding are discussed in relation to oil pollution of mangrove stands under field conditions.     

Biogeochemical cycling constraints on stream ecosystem recovery

In systems where production is limited by the availability of a nutrient, nutrient input to and recycling within the system is related to the resilience, or speed of recovery, of a system to its steady state following a disturbance. In particular, it is shown that the return timeT _s of the system to steady state, or the inverse of the resilience, is approximately equal to the mean turnover time of the limiting nutrient in the system. From this relationship, it is possible to understand and predict how various properties of food webs and their environments affect resilience. These properties include nutrient input rate, loss rate, size of the detritus compartment, and trophic structure. The effects of these properties on resilience are described by using simple mathematical models. To test model predictions, experimental studies of the response of periphyton-dominated stream ecosystems to disturbance are being conducted on a set of laboratory streams in which nutrient inputs and grazing intensity are regulated at different levels. In streams without snail grazers (low-grazed streams), 90% recirculation of stream water to reduce nutrient inputs resulted in longer turnover times (T _r ) of phosphorus within the stream compared with once-through flow. However, in streams with snail grazers (high-grazed streams), there were no differences in phosphorus turnover time between once-through and partially recirculated treatments. Results on the rate of recovery of periphyton from a flood/scour disturbance to each stream partially support the model prediction of a positive relationship between ecosystem return time (T _s ) and nutrient turnover time (T _r ) within the streams.     

Analysis of transgenic Indian mustard plants for phytoremediation of metal-contaminated mine tailings

Transgenic Indian mustard [Brassica juncea (L.) Czern.] plants overproducing the enzymes gamma-glutamylcysteine synthetase (ECS) or glutathione synthetase (GS) were shown previously to have increased levels of the metal-binding thiol peptides phytochelatins and glutathione, and enhanced Cd tolerance and accumulation. Furthermore, transgenic Indian mustard plants overexpressing adenosine triphosphate sulfurylase (APS) were shown to have higher levels of glutathione and total thiols. These results were obtained with a solution culture. To better examine the phytoremediation potential of these transgenics, a greenhouse experiment was performed in which the transgenics were grown on metal-contaminated soil collected from a USEPA Superfund site near Leadville, Colorado. A grass mixture used for revegetation of the site was included for comparison. The ECS and GS transgenics accumulated significantly (P < 0.05) more metal in their shoot than wild-type (WT) Indian mustard, while the APS plants did not. Of the six metals tested, the ECS and GS transgenics accumulated 1.5-fold more Cd, and 1.5- to 2-fold more Zn, compared with wild-type Indian mustard. Furthermore, the ECS transgenics accumulated 2.4- to 3-fold more Cr, Cu, and Pb, relative to WT. The grass mixture accumulated significantly less metal than Indian mustard: approximately 2-fold less Cd, Cu, Mn, and Zn, and 5.7-fold less Pb than WT Indian mustard. All transgenics removed significantly more metal from the soil compared with WT Indian mustard or an unplanted control. While WT did not remove more metal than the unplanted control for any of the metals tested, all three types of transgenics significantly reduced the soil metal concentration, and removed between 6% (Zn) and 25% (Cd) of the soil metal. This study is the first to demonstrate enhanced phytoextraction potential of transgenic plants using polluted environmental soil. The results confirm the importance of metal-binding peptides for plant metal accumulation and show that results from hydroponic systems have value as an indicator for phytoremediation potential.