Phytostabilisation can benefit from phytostimulatory rhizobacteria. Forty-three bacterial strains were isolated from the roots of the metallicolous legume Anthyllis vulneraria ssp. carpatica grown in a highly contaminated mine tailing (total Cd, Pb and Zn were up to 1200; 34,000; and 170,000 mg kg⁻¹, respectively). We aimed at evaluating their phytostimulatory effects on the development of leguminous metallophytes. Strains were screened for fluorescent siderophores and auxin synthesis, inorganic P solubilisation and 1-amino-cyclopropane-1-carboxylate deaminase (ACCd) activity to define a subset of 11 strains that were inoculated on the leguminous metallophytes A. vulneraria and Lotus corniculatus grown in diluted mine spoil (Zn 34,653; Pb 6842; and Cd 242, all in mg kg⁻¹). All strains were affiliated to Pseudomonas spp. (except two), synthetised auxins and siderophores and solubilised P (except three), and seven of them were ACCd positive. The inoculation effects (shoot-root-nodule biomass, chlorophyll content) depended on legume species and bacterial strain genotype. Phytostimulation scores were unrelated to siderophore/auxin synthesis and P solubilisation rates. Inoculations of the strain nos. 17–43 triggered a 1.2-fold significant increase in the chlorophyll content of A. vulneraria. Chlorophyll content and root biomass of L. corniculatus were significantly increased following the inoculations of the strain nos. 17–22 (1.5–1.4-fold, respectively). The strongest positive effects were related to increases in the nodule biomass of L. corniculatus in the presence of three ACCd-positive strains (1.8-fold), one of which was the highest auxin producer. These data suggest to focus on interactions between ACCd activity and auxin synthesis to enhance nodulation of metallicolous legumes.

Mining activities generate spoils and effluents with extremely high metal concentrations of heavy metals that might have adverse effects on ecosystems and human health. Therefore, information on soil and plant metal concentrations is needed to assess the severity of the pollution and develop a strategy for soil reclamation such as phytoremediation. Here, we studied soils and vegetation in three heavily contaminated sites with potential toxic metals and metalloids (Zn, Pb, Cd, As, TI) in the mining district of Les Malines in the Languedoc region (southern France). Extremely high concentrations were found at different places such as the Les Aviniéres tailing basins (up to 160,000 mg kg–1 Zn, 90,000 mg kg–1 Pb, 9,700 mg kg–1 of As and 245 mg kg–1 of Tl) near a former furnace. Metal contamination extended several kilometres away from the mine sites probably because of the transport of toxic mining residues by wind and water. Spontaneous vegetation growing on the three mine sites was highly diversified and included 116 plant species. The vegetation cover consisted of species also found in non-contaminated soils, some of which have been shown to be metal-tolerant ecotypes (Festuca arvernensis, Koeleria vallesiana and Armeria arenaria) and several Zn, Cd and Tl hyperaccumulators such as Anthyllis vulneraria, Thlaspi caerulescens, Iberis intermedia and Silene latifolia. This latter species was highlighted as a new thallium hyperaccumulator, accumulating nearly 1,500 mg kg–1. These species represent a patrimonial interest for their potential use for the phytoremediation of toxic metal-polluted areas.

Kidney vetch (Anthyllis vulneraria L.) is a well-known Zn hyperaccumulator. Zn often occurs with Pb in one ore; thus, plants inhabiting waste dumps are exposed not only to Zn but also to Pb toxicity. While the response of kidney vetch to Zn toxicity is relatively well known, the Pb survival strategy of Anthyllis vulneraria has not been the subject of investigations. The aim of presented research was to determine the survival strategy of kidney vetch exposed to high lead concentrations. Shoot explants of a calamine kidney vetch ecotype were placed on agar media containing 0.0, 0.5, 1.0, and 1.5 mM Pb. Morphological, physiological, and biochemical responses, in particular photosynthetic apparatus of plantlets, were examined. The most pronounced changes were observed in plants grown on media supplemented with 1.5 mM Pb after 8 weeks of culture. Increased dry weight and high lead accumulation were observed in roots. Similarly, in shoots, increased dry weight and a decreased number of newly formed shoots were recorded. The accumulation of lead was many times lower in shoots than in roots. In leaf cells’ ultra-structure, looser arrangement of chloroplast thylakoid grana was observed. Despite the decrease in chlorophyll a and carotenoid content, the photosynthetic apparatus remained efficient due to the lack of photoinhibition and increased electron transport rate beyond photosystem II (PSII). For the first time, an acclimatization mechanism based on maintaining the high efficiency of photosynthetic apparatus resulting from increasing of electron transport rate was described.

Anthyllis vulneraria was highlighted here as a Zn-hyperaccumulator for the development of a pilot phytoextraction process in the mine site of Les Avinières in the district of Saint-Laurent-Le-Minier. A. vulneraria appeared to hyperaccumulate the highest concentration of Zn in shoots with a better metal selectivity relative to Cd and Pb than the reference Zn-hyperaccumulator Noccea caerulescens. A bigger biomass production associated to a higher Zn concentration conducted A. vulneraria to the highest total zinc gain per hectare per year. As a legume, A. vulneraria was infected by rhizobia symbionts. Inoculation of A. vulneraria seeds showed a positive impact on Zn hyperaccumulation. A large-scale culture process of symbiotic rhizobia of A. vulneraria was investigated and optimized to allow large-scale inoculation process. Contaminated shoots of A. vulneraria were not considered as wastes and were recovered as Eco-Zn catalyst in particular, examples of organic synthesis, electrophilic aromatic substitution. Eco-Zn catalyst was much more efficient than conventional catalysts and allowed greener chemical processes.