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All the regulations that define a maximum concentration of metals in the receiving soil are based on total soil metal concentration. However, the potential toxicity of a heavy metal in the soil depends on its speciation and availability. We studied the effects of heavy metal speciation and availability on soil microorganism activities along a Cu/Zn contamination gradient. Microbial biomass and enzyme activity of soil contaminated with both Cu and Zn were investigated. The results showed that microbial biomass was negatively affected by the elevated metal levels. The microbial biomass-C (C(mic))/organic C (C(org)) ratio was closely correlated to heavy metal stress. There were negative correlations between soil microbial biomass, phosphatase activity and NH4NO3 extractable heavy metals. The soil microorganism activity could be predicted using empirical models with the availability of Cu and Zn. We observed that 72% of the variation in phosphatase activity could be explained by the NH4NO3-extractable and total heavy metal concentration. By considering different monitoring approaches and different viewpoints, this set of methods applied in this study seemed sensitive to site differences and contributed to a better understanding of the effects of heavy metals on the size and activity of microorganisms in soils. The data presented demonstrate the relationship between heavy metals availability and heavy metal toxicity to soil microorganism along a contamination gradient.

To study Pseudomonas putida CZ1, having high tolerance to copper and zinc on the removal of toxic metals from aqueous solutions, the biosorption of Cu(II) and Zn(II) by living and nonliving P. putida CZ1 were studied as functions of reaction time, initial pH of the solution and metal concentration. It was found that the optimum pH for Zn(II) removal by living and nonliving cells was 5.0, while it was 5.0 and 4.5, respectively, for Cu(II) removal. At the optimal conditions, metal ion biosorption was increased as the initial metal concentration increased. The adsorption data with respect to both metals provide an excellent fit to the Langmuir isotherm. The binding capacity of living cells is significantly higher than that of nonliving cells at tested conditions. It demonstrated that about 40-50% of the metals were actively taken up by P. putida CZ1, with the remainder being passively bound to the bacterium. Moreover, desorption efficiency of Cu(II) and Zn(II) by living cells was 72.5 and 45.6% under 0.1M HCl and it was 95.3 and 83.8% by nonliving cells, respectively. It may be due to Cu(II) and Zn(II) uptake by the living cells enhanced by intracellular accumulation.

The role of rhizosphere bacteria in facilitating the solubility of copper (Cu) in contaminated soil and Cu accumulation in plant were studied. The bacteria strains were isolated from the rhizosphere of Elsholtzia splendens, a Cu accumulator growing on Tonglu Mountain copper mines. After the sandy soils containing 237 mg kg(-1) were incubated with the bacteria strains, it was indicated that rhizosphere microbes played an important role in influencing the availability of water-soluble Cu in soils. Soils had greater concentrations of water-extractable Cu compared with axenic soils inoculated with different bacterial strains. Further evidence for bacterial facilitation of increased solubility of Cu in the soil was obtained using the antibiotic ampicillin (0.1 mg g(-1)). There were 36% decreases in Cu concentration in the presence of bacterial strain MS12 and ampicillin together compared with bacterial inoculation alone. Different bacterial strains had different abilities on soil water-soluble Cu. To achieve the highest rates of plant Cu accumulation, it was necessary for bacteria to be present in the rhizosphere of E. splendens. Inoculated plants supplied with 20 micromol L(-1) CuSO4 had significantly greater concentrations of Cu in shoots and roots than uninoculated plants and bacterial strain MS2 was the most effective strain in promoting plant Cu uptake. There were 2.2-fold and 2.5-fold increases in Cu accumulation in the shoots and roots of plants inoculated with strain MS2 compared to axenic controls. Furthermore, when ampicillin and the bacterial strains were added together to the nutrient solution, the Cu concentrations in roots and shoots of ampicillin-treated plants were lower than those in inoculated plants. When ampicillin was added to the nutrient solution, Cu accumulation was inhibited by about 24-44% in shoots and 20-44% in roots. The above results provided a new insight into the phytoremediation of Cu-contaminated soil.

We studied the effects of copper (Cu) phytoremediation on microbial community composition under laboratory conditions. A Cu accumulator, Commelina communis, was grown on a soil containing different gradients of Cu. Results showed that the biomass of C. communis grown with Cu differed from that of the control. Concentrations of Cu in the shoots of C. communis were 73.6, 160.9, and 319.1 mg kg-1 under 200, 500, and 1000 mg kg-1 Cu treatments, respectively. Polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) fingerprint analysis revealed that additions of Cu decreased the number of bands in the soil with C. communis or without plants. The principal component analysis explained 52.7% of the variance for the different rhizospheres of soil and Cu treatments in the soil samples. These results indicated significant effects on soil bacteria activity and community composition in the rhizosphere of C. communis and provided a basis for further studies of metal-accumulator plant effects on soil microorganisms. [ABSTRACT FROM AUTHOR]