The aim of this work was to test Lupinus luteus plants, inoculated with metal resistant rhizobacteria, in order to phytostabilise metals in contaminated soils. The resistance to heavy metals of strains isolated from nodules of Lupinus plants was evaluated. The strain MSMC541 showed multi-resistance to several metals (up to 13.3 mM As, 2.2 mM Cd, 2.3 mM Cu, 9 mM Pb and 30 mM Zn), and it was selected for further characterization. Furthermore, this strain was able to biosorb great amounts of metals in cell biomass. 16S rDNA sequencing positioned this strain within the genus Serratia. The presence of arsenic resistance genes was confirmed by southern blot and PCR amplification. A rhizoremediation pot experiment was conducted using Lupinus luteus grown on sand supplemented with heavy metals and inoculated with MSMC541. Plant growth parameters and metal accumulation were determined in inoculated vs. non-inoculated Lupinus luteus plants. The results showed that inoculation with MSMC541 improved the plant tolerance to metals. At the same time, metal translocation to the shoot was significantly reduced upon inoculation. These results suggest that Lupinus luteus plants, inoculated with the metal resistant strain Serratia sp. MSMC541, have a great potential for phytostabilization of metal contaminated soils.
Four strains of rhizobia nodulating Acacia were isolated from the Moroccan desert soil by trapping with seedlings of Acacia gummifera and Acacia raddiana, and were studied for their ability to tolerate high salinity and dryness conditions. The strains MDSMC 2, MDSMC 18 and MDSMC 50 were halotolerant (they tolerated up to 1 M NaCl) and they accumulated glutamate and mannosucrose. The synthesis of the latter solute, which is the major endogenous osmolyte, is partially repressed in the presence of glycine betaine. The strain MDSMC 34 was less halotolerant (growth inhibited by a concentration greater than 0.5 M NaCl), and accumulated trehalose (as the main endogenous osmolyte) and glutamate. Rhizobia from the Moroccan desert soil were highly resistant to desiccation and their tolerance to dryness was stimulated by osmotic pretreatment. Thus, the accumulation of mannosucrose or trehalose by desert rhizobia represents both an osmoadaptative response and a part of a desiccation tolerance mechanism.
Two strains of Rhizobia isolated from nodules of Vicia faba var. major and one strain isolated from nodules of Cicer arietinum L. were characterized for salt resistance. The presence of 1 mM glycine betaine or choline in a minimal medium with added NaCl had a beneficial role on the growth of the three strains. Both molecules were found to be taken up by cells obtained at low osmolarity, and whereas glycine betaine uptake activity was stimulated significantly in cells grown in the presence of 0.15 M NaCl, choline uptake activity was strongly inhibited by salt in all tested strains. However, in cells grown with exogenous choline,the uptake inhibition exerted by salt was relieved, mainly in the strain isolated from nodules of C. arietinum L. On the basis of kinetics determinations, in control cells as well as in salt-stressed cells, only high-affinity activities were observed for glycine betaine and choline(apparent Kms between 3 and 18 micro;M). Periplasmic proteins that bound glycine betaine or choline were identified. In nondenaturing conditions, these proteins extracted from the various strains showed different electrophoretic mobility with always a less negative entire charge than the analogous proteins from Rhizobium meliloti.
Rhizosphere bacterial populations associated with four metallophyte plants in one of major polymetallic (Pb-Zn-Cu) semiarid Moroccan Hercynian province (DraaˆSfar, Marrakech, Morocco) presenting long-term contamination mainly with Zn and Pb were analysed and compared to selected control soils. In the highly Zn-, Cu-, Pb-and Cdcontaminated soils, the total number of culturable heterotrophic bacteria were found in low proportions (<2.6 · 10 2 -1.6 · 10 4 g )1 soil). This bacterial content was slightly similar to that found in moderately polluted and controls soils (6.7 · 10 4 -5.8 · 10 6 ). However, the bacterial diversity and the rhizosphere/soil ratio, which compares the bacterial content (or bacterial charge) around the metallophyte plants with that in non-rhizosphere soil, were the bacteriological parameters mostly affected by heavy metal contamination. The chronic Zinc-stress results in an increase of tolerance to this metal of both the rhizosphere and non-rhizosphere bacterial communities. However, in general, the rhizosphere bacterial populations exhibited less tolerance to Zn toxicity than the bacterial population of non-rhizosphere soils. This result suggests that toxic effects of Zn decrease in the rhizosphere soils of the metallophyte plants.
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