Abstract
Two legumes (Anthyllis vulneraria and Lotus corniculatus) adapted to heavy metals form an interesting material for phytostabilisation strategy in mining sites. As biological nitrogen fixators, these legumes associated with compatible symbiotic bacteria provide an efficient establishment of a sustainable cover vegetation limiting metal dispersion in the environment. Our objectives were to study the effects of heavy metals and the host plant on symbiotic populations naturally associated with these legumes by analyzing (i) symbiotic populations associated with A. vulneraria on 8 contaminated and uncontaminated sites (ii) rhizobial populations associated with L. corniculatus that were compared with those of Anthyllis. The distribution of mesorhizobial strains isolated from A. vulneraria root-nodules from several contaminated and uncontaminated sites depends on high levels of heavy metals in soils by selecting highly resistant strains and impacting the taxonomic composition. Strains belonging to M. metallidurans were only found in highly contaminated sites. Two new potential metal-tolerant species were detected in two distinct mines. One of them was closely related to M. ciceri and M. loti and its members had the feature of not -possessing the cadA gene, a gene involved in metal-tolerance among M. metallidurans strains. By contrast, uncontaminated sites revealed a different taxonomic diversity with new species sensitive to heavy metals. Four of these new species were defined. A. vulneraria and L. corniculatus share the same taxonomic diversity in the contaminated sites tested. By contrast, symbiotic properties of the strains vary depending on the host plant used for trapping. Strains belong either to symbiovar (sv.) anthyllidis or to sv. loti according to geographic origins and independently of heavy metal levels in soils. A. vulneraria associated with strains of sv. anthyllidis or sv. loti. In contrast, L. corniculatus only associated with strains of sv. loti. In contaminated or uncontaminated soils, A. vulneraria was preferentially nodulated by sv. anthyllidis. In conclusion, A. vulneraria and L. corniculatus established symbiotic relationships with the same taxonomic groups of Mesorhizobium but associated with different symbiovars. The finding of taxonomic groups strongly depends on geographical sites, suggesting special adaptations to environmental conditions. Use of local biological resources is the strategy we recommend for revegetation of old mines.