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Tailings from the Zeïda mining region, located in the Middle Atlas Mountains of Morocco, contain high levels of lead and zinc which have many adverse effects, regarding both the environment and the health of the local human population. Finding practical methods to limit heavy metal dispersion and subsequent pollution of ecosystems in this area is therefore critical. This study aims to evaluate lead-tolerant rhizobacteria with an aim of exhibiting multiple plant growth-promoting traits. Thus, the growth of Medicago sativa may be improved and its resistance under lead stress conditions and may be subsequently used for the phytostabilization of lead-contaminated soils. Forty bacteria were isolated from the rhizospheric soil of Astragalus armatus plants growing wildly in the Zeïda mine tailings. After preventing the duplicates of obtained isolates, the resistance to various heavy metals at high levels allowed the selection of two strains (i.e. AaR114 and AaR72). These strains were evaluated in vitro for characteristics that promote plant development, such as the synthesis of 1-aminocyclopropane-1-carboxylic acid deaminase, indoleacetic acid, hydrogen cyanide, siderophore, phosphate solubilization, and antifungal activity. Inoculation of M. sativa plants with rhizobacteria AaR114 and AaR72, in the presence of 100 μg mL−1 of lead-acetate, was shown to significantly improve plant tolerance, increase aerial and root biomass, and diminished the negative impacts of heavy metals on plants. The 16S rRNA sequences analyses of the bacteria revealed that the strains AaR114 and AaR72 were linked to Bacillus subtilis DSM 10 T and Neobacillus niacini NBRC 15566 T, respectively. [ABSTRACT FROM AUTHOR]

The present paper aims to study the phytoremediation of a polymetallic abandoned mine site in the northwest of Marrakesh, Morocco, by exploring the potential of the symbiotic relationship between Pinus halepensis and the ectomycorrhizal fungi belonging to the Scleroderma genus. This process was combined with the use of sand to neutralize the acidic mine tailings (pH < 3) and to stabilize the heavy metals. Inoculated and non-inoculated plants were grown under greenhouse conditions in a substrate containing different proportions of mine tailings: 0%, 10%, 25%, 50%, 75% and 100% mixed with sand as an amendment. After a growth period of six months, the results showed a significant increase (p ≤ 0.05) of the pH (11%) and the electrical conductivity of the substrate (29%). Moreover, these treatments significantly increased (p ≤ 0.05) the amounts of metallic trace elements, lead (39%), cadmium (22%) and zinc (28%) in the roots of inoculated seedlings compared to non-inoculated ones. The bioaccumulation factor reached a value higher than 1, and the translocation factor recorded a value below 1 particularly for zinc in all treatments and for copper and lead in the 25%, 50% and 75% treatments, indicating that Pinus halepensis has the ability to accumulate significant concentrations of these metals in its roots, thus making a suitable plant for phytoremediation or phytostabilization programs. The results of this study also revealed that the mine tailing amendment had a significant impact on the efficiency of the phytoremediation process toward the metals detected in these polluted soils. [ABSTRACT FROM AUTHOR]

Mining activity in the Touissit district of Eastern Morocco has led to an unprecedented accumulation of heavy metals, mainly lead and zinc, in the tailing ponds of the open-air mines. This poses a real danger to both the environment and local population. The goal of this work was to characterize the P lant G rowth P romoting R hizobacteria (PGPR) isolated from the rhizosphere soil of R. pseudoacacia plants grown wild in the abandoned Pb- and Zn-contaminated tailing ponds in the mining district of Touissit, in Eastern Morocco. One hundred bacterial strains were isolated from the rhizosphere of black locust (Robinia pseudoacacia L.) plants growing naturally in the Touissit mine tailings. Quantitative determination of indole-acetic and siderophores production, inorganic phosphate solubilization, hydrolysis of 1-aminocyclopropane-1-carboxylic acid (ACC deaminase activity), and ability to act as a biocontrol agent allowed selection of the 3 strains, 7MBT, 17MBT and 84MBT with improved PGP properties. The three strains grew well in the presence of high concentration of Pb-acetate and ZnCl 2; and the addition of Pb or Zn to the culture medium differently affected the PGP properties analyzed. Inoculation of black locust grown with the 3 selected strains, in the presence 1000 μg ml−1 of Pb-acetate, produced varying effects on the plant dry weight. The strain 84MBT alone or in combination with strains 7MBT and 17MBT increased significantly the dry weight of the plants by 91, 62, and 85% respectively. The 16S rRNA gene sequence analysis of each strain showed that the strains 7MBT 17MBT and 84MBT had 99.34, 100, and had 99.72% similarity with Priestia endophytica (formerly B. endophyticus), B. pumilus NBRC 12092T, and B. halotolerans NBRC 15718T, respectively. • Out of 100 strains, three were characterized as the best preforming in the PGP activities. • The three strains were identified as Bacillus pumilus, B. endophyticus, and B. halotolerans. • The strains tolerate high lead and zinc concentrations. • The metal stress enhanced the PGP activities of the three strains. • The inoculation by the strains improved significantly the growth of black locust by 85–91%. [ABSTRACT FROM AUTHOR]

The abandoned Kettara pyrrhotite mine (Marrakech region, Morocco) is a real source of acid mine drainage (AMD) and heavy metal pollution from previous mining operations—which has spread, particularly because of wind erosion. A store-and-release cover system made of phosphate wastes was built on the site for preventing AMD. To ensure the integrity of this cover and its durability, it is desirable to revegetate it (phytostabilization) with plants adapted to the edaphoclimatic conditions of the region. In this paper, a study was carried out on the spontaneous vegetation around the phosphate cover in order to consider the selection of plants to promote the stabilization of the Kettara mine tailings pond. Nine species of native plants with their rhizospheric soils growing in agricultural soils and tailings from the Kettara mine were collected, and metals (As, Cd, Co, Cu, Pb, Zn, Ni, Cr) were analyzed. The soil analysis showed that the tailings contained high concentrations of Cu (177.64 mg/kg) and Pb (116.80 mg/kg) and that the agricultural soil contained high concentrations of As (25.07 mg/kg) and Cu (251.96 mg/kg) exceeding the toxicity level (Cu > 100 mg/kg, Pb > 100 mg/kg, As > 20 mg/kg). The plant analysis showed low trace metal accumulation in Scolymus hispanicus, Festuca ovina, Cleome brachycarpa, Carlina involucrata and Peganum harmala. These species had a bioconcentration factor (BCF) greater than 1 and a translocation factor (TF) less than 1, demonstrating a high tolerance to trace metals. Therefore, they are good candidates for use in the phytoremediation of the Kettara mine tailings. These species could also potentially be used for the phytostabilization of the phosphate waste cover of the Kettara mine, thus completing the rehabilitation process of this area. [ABSTRACT FROM AUTHOR]

The management of mine waste has become an urgent issue, especially in semi-arid environments. In this context, and with an aim to inhibit the oxidation of the sulfide tailings of the abandoned mine of Kettara in Morocco, a store-and-release (SR) cover made of phosphate mine waste (PW) was implemented. In order to guarantee its long-term performance, phytostabilization by local wild plant species is currently the most effective and sustainable solution. This study aimed to assess the growth performance and phytostabilization efficiency of five local wild plant species to grow on the SR cover made of PW. A field experiment was conducted for two growing seasons (2018 and 2019), without amendments and with the minimum of human care. PW and the aboveground and belowground parts of the studied plant species were collected and analyzed for As, Cd, Cu, Ni, and Zn. The bioconcentration factor (BCF) and translocation factor (TF) were also calculated. Despite the hostile conditions of the mining environment, the five plant species showed promising growth performances as follows: Atriplex semibaccata > Vicia sativa > Launaea arborescens > Peganum harmala > Asparagus horridus. The five plants showed high accumulation capacity of the trace elements, with the highest concentrations in belowground tissue. Principal component analysis distinguished A. semibaccata as having a high concentration of Cu and As, while Asparagus horridus had higher concentrations of Cd and Zn. In contrast, P. harmala, V. sativa, and L. arborescens demonstrated affinity regarding Ni. According to the BCF (<1) and TF (<1), these plant species could be used as effective phytostabilizers of the studied trace elements. The present study showed that local wild plant species have a great potential for the phytostabilization of PW, and could ensure the long-term efficiency of SR cover. [ABSTRACT FROM AUTHOR]

The aim of this work was to characterize and identify some bacteria isolated from the root nodules of Retama monosperma grown in Sidi Boubker lead and zinc mine tailings. Very few root nodules were obtained on the root nodules of R. monosperma grown in these soils. The three bacteria isolated from the root nodules were tolerant in vitro to different concentrations of heavy metals, including lead and zinc. The rep-PCR experiments showed that the three isolates have different molecular fingerprints and were considered as three different strains. The analysis of their 16S rRNA gene sequences proved their affiliation to the genus Bradyrhizobium. The analysis and phylogeny of the housekeeping genes atpD, glnII, gyrB, recA, and rpoB confirmed that the closest species was B. valentinum with similarity percentages of 95.61 to 95.82%. The three isolates recovered from the root nodules were slow-growing rhizobia capable to renodulate their original host plant in the presence of Pb-acetate. They were able to nodulate R. sphaerocarpa and Lupinus luteus also but not Glycine max or Phaseolus vulgaris. The phylogeny of the nodA and nodC nodulation genes as well as the nifH gene of the three strains showed that they belong to the symbiovar retamae of the genus Bradyrhizobium. The three strains isolated could be considered for use as inoculum for Retama plants before use in phytoremediation experiments.

Screening of native plant species from mining sites can lead to identify suitable plants for phytoremediation approaches. In this study, we assayed heavy metals tolerance and accumulation in native and dominant plants growing on abandoned Pb/Zn mining site in eastern Morocco. Soil samples and native plants were collected and analyzed for As, Cd, Cu, Ni, Sb, Pb, and Zn concentrations. Bioconcentration factor (BCF), translocation factor (TF), and biological accumulation coefficient (BAC) were determined for each element. Our results showed that soils present low organic matter content combined with high levels of heavy metals especially Pb and Zn due to past extraction activities. Native and dominant plants sampled in these areas were classified into 14 species and eight families. Principal components analysis separated Artemisia herba-alba with high concentrations of As, Cd, Cu, Ni, and Pb in shoots from other species. Four plant species, namely, Reseda alba, Cistus libanotis, Stipa tenacissima, and Artemisia herba-alba showed strong capacity to tolerate and hyperaccumulate heavy metals, especially Pb, in their tissues. According to BCF, TF, and BAC, these plant species could be used as effective plants for Pb phytoextraction. Stipa tenacissima and Artemisia herba-alba are better suited for phytostabilization of Cd/Cu and Cu/Zn, respectively. Our study shows that several spontaneous and native plants growing on Pb/Zn contaminated sites have a good potential for developing heavy metals phytoremediation strategies. [ABSTRACT FROM AUTHOR]

This study uses an ecotoxicology approach to evaluate the effectiveness of combining powdered marble as an amendment, with phytostabilization by Medicago sativa L. on the neutralization of acidic mine tailings, and the stabilization of heavy metals. The mine tailings were collected from an abandoned polymetallic mine in Southern Morocco, and mixed with powdered marble as the following proportions, 25%, 50%, and 75%. Laboratory immobilization/stabilization tests showed that the application of powdered marble in the treatments led to a significant increase in pH, and significant reductions of Cu, Zn (99%), Pb (98%), and Fe (45%). Greenhouse experiments showed that plant growth in all treatments was significantly (p ≤ 0.05) less than growth in agricultural soil. Plant growth significantly (p ≤ 0.05) decreased as the proportion of powdered marble increased. The concentration of metals in plant roots were significantly (p ≤ 0.05) higher than those of shoots. Combining immobilization by powdered marble with phytostabilization by M. sativa L. could represent a viable method of rehabilitating acidic polymetallic mine tailings.

Waste dumps generated by mining activities contain heavy metals that are dispersed into areas leading to significant environmental contamination. The objectives of this study were (i) to survey native plants and their associated AM fungal communities from waste soils in a Moroccan mine site and (ii) to follow Eucalyptus growth in soil collected from the waste-mine. AM spores from native plant species were collected from the mining site and the surrounding uncontaminated areas were multiplied and inoculated onto Eucalyptus camaldulensis. The results showed that (i) the native plant species recorded in the waste did not show an active metal uptake, (ii) the selected native plant species are associated with AM mycorrhizal fungi and (iii) the use of AM fungi adapted to these drastic conditions can improve the growth of the fast-growing tree, E. camaldulensis and its tolerance to high soil Cu content. In conclusion, it is suggested that in order to define efficient low-cost phytostabilization processes, the use of native resources (i.e., mixtures of native mycorrhizal fungi) in combination with fast-growing tree species such as Eucalyptus, could be used to optimize the establishment of a permanent cover plant in contaminated areas.