Assisted Phytoremediation between Biochar and Crotalaria pumila to Phytostabilize Heavy Metals in Mine Tailings.

The increasing demand for mineral resources has generated mine tailings with heavy metals (HM) that negatively impact human and ecosystem health. Therefore, it is necessary to implement strategies that promote the immobilization or elimination of HM, like phytoremediation. However, the toxic effect of metals may affect plant establishment, growth, and fitness, reducing phytoremediation efficiency. Therefore, adding organic amendments to mine tailings, such as biochar, can favor the establishment of plants, reducing the bioavailability of HM and its subsequent incorporation into the food chain. Here, we evaluated HM bioaccumulation, biomass, morphological characters, chlorophyll content, and genotoxic damage in the herbaceous Crotalaria pumila to assess its potential for phytostabilization of HM in mine tailings. The study was carried out for 100 days on plants developed under greenhouse conditions under two treatments (tailing substrate and 75% tailing/25% coconut fiber biochar substrate); every 25 days, 12 plants were selected per treatment. C. pumila registered the following bioaccumulation patterns: Pb > Zn > Cu > Cd in root and in leaf tissues. Furthermore, the results showed that individuals that grew on mine tailing substrate bioaccumulated many times more metals (Zn: 2.1, Cu: 1.8, Cd: 5.0, Pb: 3.0) and showed higher genetic damage levels (1.5 times higher) compared to individuals grown on mine tailing substrate with biochar. In contrast, individuals grown on mine tailing substrate with biochar documented higher chlorophyll a and b content (1.1 times more, for both), as well as higher biomass (1.5 times more). Therefore, adding coconut fiber biochar to mine tailing has a positive effect on the establishment and development of C. pumila individuals with the potential to phytoextract and phytostabilize HM from polluted soils. Our results suggest that the binomial hyperaccumulator plant in combination with this particular biochar is an excellent system to phytostabilize soils contaminated with HM.

Multi-biomarker approach reveals the effects of heavy metal bioaccumulation in the foundation species Prosopis laevigata (Fabaceae).

Mining is a major economic activity in many developing countries. However, it disturbs the environment, producing enormous quantities of waste, known as mine tailings, which can have deleterious environmental impact, due to their high heavy metals (HM) content. Often, foundation species that establish on mine tailings are good candidates to study the effects of HM bioaccumulation at different levels of biological organization. Prosopis laevigata is considered a HM hyperaccumulator which presents attributes of a foundation species (FS) and establishes naturally on mine tailings. We evaluated the bioaccumulation of Cu, Pb, and Zn in P. laevigata foliar tissue, the leaf micro- and macro-morphological characters, DNA damage, and population genetic effects. In total, 80 P. laevigata individuals (20/site) belonging to four populations: The individuals from both sites (exposed and reference) bioaccumulated HMs (Pb > Cu > Zn). However, in the exposed individuals, Pb and Cu bioaccumulation was significantly higher. Also, a significant effect of macro- and micro-morphological characters was registered, showing significantly lower values in individuals from the exposed sites. In addition, we found significant differences in genotoxic damage in P. laevigata individuals, between the exposed and reference sites. In contrast, for the micro-morphological characters, none of the analyzed metals had any influence. P. laevigata did not show significant differences in the genetic structure and diversity between exposed and reference populations. However, four haplotypes and four private alleles were found in the exposed populations. Since P. laevigata is a species that establishes naturally in polluted sites and bioaccumulates HM in its foliar tissues, the resulting genetic, individual and population effects have not been severe enough to show detrimental effects; hence, P. laevigata can be a useful tool in phytoremediation strategies for soils polluted with Pb and Cu, maintaining its important ecological functions.

The bioaccumulation potential of heavy metals by Gliricidia sepium (Fabaceae) in mine tailings.

As a result of mining activities, waste of different types is generated. One example is mine tailings that contain potentially toxic elements such as heavy metals that negatively impact the environment and human health. Hence, developing treatments to guarantee its efficient elimination from the environment is necessary. Among these treatments, phytoremediation takes advantage of the potential of different plant species, to remove heavy metals from polluted sites. Gliricidia sepium is a tree that grows up to 15 m high and distributed from southern Mexico to Central America. This study evaluates the heavy metal bioaccumulation capacity in roots and leaves, and the effect of such bioaccumulation on fifteen macro- and one micro-morphological characters of G. sepium growing during 360 days in control, and in mine tailing substrates. G. sepium individuals growing on the exposed substrate registered the following average heavy metal bioaccumulation pattern in the roots: Fe > Pb > Zn > Cu, while in the leaf tissue, the bioaccumulation pattern was Cu > Fe > Pb > Zn. Macro- and micro-morphological characters evaluated in G. sepium decreased in plants exposed to metals. The translocation factor showed that Cu and Pb registered average values greater than 1. In conclusion, G. sepium is a species with potential for the phytoremediation of soils contaminated with Fe, Cu, and Pb, and for phytostabilizing soils polluted with Fe, Pb, Zn, and Cu, along with its ability to establish itself and turn into an abundant plant species in polluted sites, its capacity to bioaccumulate heavy metals in roots and leaves, and its high rate of HM translocation.

Anoxybacillus sp. Strain UARK-01, a New Thermophilic Soil Bacterium with Hyperthermostable Alkaline Laccase Activity.

We describe the isolation and characteristics of a novel thermophilic bacterium from soil. The organism is a member of the Anoxybacillus genus based on phylogenetic analysis of the 16S rRNA gene. The 16S rRNA of the organism shares >99% sequence identity with those of two species, Anoxybacillus rupiensis and A. geothermalis. We named this isolate as Anoxybacillus sp. strain UARK-01. UARK-01 grows optimally in the presence of oxygen at 55 °C and pH 8. It grew excellently in the presence of lignin as the sole carbon source. Culture supernatant from UARK-01 grown on lignin was rich in laccase activity. The laccase activity was optimal at 90 °C and pH 9, and there was comparable activity at 80 and 100 °C. The crude laccase decolorized approximately 75% of Congo Red in 7 h under optimal conditions. A single laccase gene was identified from the draft genome sequence of Anoxybacillus sp. UARK-01. The UARK-01 laccase (Anox_Lacc) was cloned and overexpressed in Escherichia coli and was partially purified. The partially purified Anox_Lacc decolorized approximately 1.64+/0.21 nanomoles of Congo Red per microgram protein in 30 min at 90 °C and pH 9. Anox_Lacc is a member of the multicopper polyphenol oxidoreductase laccase family (pfam02578 Cu-oxidase_4) and has novel characteristics. Multiple sequence analysis of Anox_Lacc with six homologs from the family revealed four conserved copper ligands and several new residues that are fully conserved. Anox_Lacc is enriched in leucine, glutamine, and lysine, and it contains fewer alanine, arginine, glycine, and serine residues. Skewed amino acid composition of Anox_Lacc likely contributes to the exceptional thermochemical properties of the laccase activity from UARK-01. Both lignin utilization and production of hyperthermostable alkaline laccase are new findings in the Anoxybacillus genus.