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Tolerance of Bradyrhizobium and Azorhizobium strains and isolates to copper, cadmium and zinc "in vitro"

Two experiments were carried out at the Soil Science Department of the Federal University of Lavras (MG), to assess the tolerance to heavy metals "in vitro" of inoculant strains (I), and isolates from both heavy metals contaminated soil (ICS) and uncontaminated soil (IUS) of Bradyrhizobium from Enterolobium contortisiliquum and Acacia mangium and of Azorhizobium from Sesbania virgata. In the first experiment ten strains and, or, isolates of each plant species, were tested in YMA media modified by addition of biological buffers (HEPES and MES), supplied with different concentrations of Cu, Cd and Zn. Copper and Cd were tested at concentrations from 0 to 40 mg L-1 for both genera, whereas Zn varied from 0 to 1.000 mg L-1 for Bradyrhizobium and from 0 to 500 mg L-1 for Azorhizobium. Growth at the different metal concentrations was evaluated with attribution of values to patterns observed (0 to 5). The ICS of both genera were more tolerant but Zn tolerance to Bradyrhizobium (800 mg L-1) and Cu (40 mg L-1) was, respectively, two and eight times higher than that of Azorhizobium. In the second experiment, strains and isolates tolerant (T), sensitive (S) and medium tolerant (MT) to metals selected in modified YMA media, were studied in aqueous solutions with different concentrations of Cu (0 to 0.01 mg L-1) Cd and Zn (0 to 1.0 mg L-1). The evaluation of the number of viable cells in metallic solutions was accomplished by counting colony forming units at 0, 24, 48, 72 and 96 h of incubation, by the method of successive dilutions and inoculation in YMA. Although the metal solutions have been more selective than modified YMA media, the two methods showed that: (1) Azorhizobium was more sensitive than Bradyrhizobium, (2) ICS of both genera were more tolerant than IUS and (3) the order of toxicity of heavy metals was Cu > Cd > Zn.

rhizobia; heavy metals; contamination of soil; symbionts


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