Alicia Margarita Godeas

Mining activities cause strong soil alterations, such as heavy metal (HM) pollution, which decreases the diversity of plant communities and rhizospheric microorganisms, including arbuscular mycorrhizal (AM) fungi. The polymetallic Paramillos de Uspallata mine in the Andes Mountains, the first mining exploitation in Argentina, provides a unique scenario to study AM fungal resilience after long-term disturbance following over 40 years of inactivity. This study aimed to analyze mycorrhizal status and AM fungal communities in the mine and a nearby unexploited area and to evaluate their associations with soil parameters to elucidate life history strategies. Long-term exposure to elevated Fe, Pb, Zn, and Ag concentrations and high electrical conductivity (EC, 5.46 mS/cm) led to the dominance of Entrophospora infrequens in association with Pappostipa speciosa, demonstrating that this AM species is a stress-tolerant strategist in symbiosis with a pioneer perennial plant, resilient in the most impacted mine areas. In contrast, the unexploited area, with an EC of 0.48 mS/cm and low HM contents, supported competitive and ruderal species, revealing distinct ecological strategies of AM fungi in disturbed versus undisturbed environments. These findings highlight the potential of E. infrequens for bioremediation and ecological restoration in post-mining landscapes.

Contaminated soils are a challenge for implementing biotechnology in bioremediation, the recovery of Critical and Strategic Raw Materials (CRMs and SRMs), and food security. European Union (EU) Governments have established strict limits on As, Pb, Cd, and Hg in foods (Document 32023R0915) and requested the recovery of 34 CRMs within a circular economy (CE) (5th CRMs list). This study proposed a biotechnological tool for the decontamination of soil with heavy metal(loid)s by arbuscular mycorrhizal (AM)-assisted phytoextraction and the subsequent recovery of CRMs or by phytostabilization to prevent their entry into the food chain. It consisted of placing Baccharis salicifolia plants, inoculated or non-inoculated with AM fungi, into bioreactors (BRs) containing mining soil with Cd, Ni, and Cu, according to the Argentinian Patent (AR090183B1). The bioextractive potential (BP) was also estimated at the highest Technological Readiness Level (TRL) using a vegetable depuration module (VDM, TRL 6). Inoculated plants showed significantly higher aerial bioaccumulation coefficients (Cd: 68.62; P: 2.99; Ni: 2.51; Cu: 0.18) in BRs, and the BP values reached 1.16 g, 9.75 g, 2.40 g, and 213.1 g for Ni, Cd, Cu, and P, respectively. Finally, these CRMs and SRMs could be recovered from biomass through hydrometallurgy within a CE framework.

Contaminated soils are a challenge for implementing biotechnology in bioremediation, recovery of Critical and Strategic Raw Materials (CRMs and SRMs), and food security. European Union (EU) Governments have established strict limits on As, Pb, Cd, and Hg in foods (Document 32023R0915), and requested the recovery of 34 CRMs within a circular economy (CE) (5th CRMs list). This study proposed a biotechnological tool for the decontamination of soil with heavy metal(loids) by arbuscular mycorrhizal (AM)-assisted phytoextraction and subsequent recovery of CRMs, or by phytostabilization to prevent their entry into the food chain. It consisted of placing Baccharis salicifolia plants, inoculated or non-inoculated with AM fungi, into Bioreactors (BRs) containing mining soil with Cd, Ni, and Cu, according to the argentinian Patent (AR090183B1). The bioextractive potential (BP) was also estimated at the highest Technological Readiness Level (TRL) using a Vegetable Depuration Module (VDM, TRL 6). Inoculated plants showed significantly higher aerial bioaccumulation coefficients (Cd: 68.62; P: 2.99; Ni: 2.51; Cu: 0.18) in BRs, and the BP values reached 1.16 g, 9.75 g, 2.40 g, and 213.1 g for Ni, Cd, Cu, and P, respectively. Finally, those CRMs and SRMs could be recovered from biomass through hydrometallurgy within a CE framework.

In recent years, critical and secondary raw materials (CRMs and SRMs, respectively) have received great interest within the circular economy model. In this work, the mycorrhizal-assisted phytomining (MAP) system, composed of Helianthus annuus–arbuscular mycorrhizal fungus Rhizophagus intraradices–Zn-volcanic ashes, was applied in bioreactors for the recovery of CRMs (Sr, P) and SRMs (Cr, Zn, Cu, Mn, Rb, Ni) from mining wastes of the Los Cóndores mine (Argentina). Our results showed high bioaccumulation of Sr, P, Mn, and Zn in the aerial tissues, and a high root-to-shoot translocation for Mn (4.02) > Sr > P > Rb > Zn (0.84). Mycorrhization treatment increased the root-to-leaf translocation for Cr and P and prevented translocation towards flower tissues in most elements. The estimated bioextracting potential of the MAP system (290 plants) in a vegetable depuration module (VDM) ranged from 158 mg/m³ P > Zn > Mn > 15.1 mg/m³ Sr. We demonstrated a promising and cost-effective biotechnology applicable in agronomical practices, given the exclusion of toxic elements in flower parts, as well as for the recovery of CRMs and SRMs by hydrometallurgy from plant biomass.

In this work, mycorrhizal-assisted phytoextraction (MAP, Helianthus annuus–arbuscular mycorrhizal fungus Rhizophagus intraradices–Zn-volcanic ashes) was applied for the recovery of secondary and critical raw materials (SRMs and CRMs, respectively) from Joda West (Odisha, India) mine residues, within a novel multidisciplinary management strategy. Mine residues were preliminarily characterized by using advanced analytical techniques, and subsequently mapped, classified and selected using multispectral satellite Sentinel-2A images and cluster analysis. Selected mine residues were treated by MAP at laboratory scale, and the fate of several SRMs (e.g., Zn, Cr, As, Ni, Cu, Ca, Al, K, S, Rb, Fe, Mn) and CRMs (such as Ga, Ti, P, Ba and Sr) was investigated. Bioconcentration factors in shoots (BC_S) and roots (BC_R) and translocation factors (TF) were: 5.34(P) > BCS > 0.00(Al); 15.0(S) > BCR > 0.038(Ba); 9.28(Rb) > TF > 0.02(Ti). Results were used to predict MAP performance at larger scale, simulating a Vegetable Depuration Module (VDM) containing mine residues (1 m³). Estimated bio-extracting potential (BP) was in the range 2417 g/m³ (K) > BP> 0.14 g/m³ (As), suggesting the eventual subsequent recovery of SRMs and CRMs by hydrometallurgical techniques, with final purification by selective electrodeposition, as a viable and cost-effective option. The results are promising for MAP application at larger scale, within a circular economy-based approach.

Los helechos presentan dos etapas en su ciclo de vida, una fase esporofítica y una gametofítica. Estos por lo general pueden colonizar diferentes ambientes y frecuentemente presentan raíces micorrizadas. Este estudio describe la asociación experimental entre Pteris vittata, un helecho leptosporangiado y una cepa de Glomus intraradices durante el ciclo de vida del helecho, desde la germinación de las esporas hasta el desarrollo del esporofito maduro. El objetivo de este estudio fue comparar los patrones de colonización de G. intraradices a lo largo de todo el ciclo de vida del helecho con los tipos encontrados en la naturaleza. Las esporas maduras fueron obtenidas de frondes fértiles de P. vittata que crecen sobre las paredes de la ciudad de Buenos Aires, Argentina. Las raíces se tiñeron y fueron observadas bajo microscopio óptico para el estudio de la colonización micorrízica. Aproximadamente 75 esporas de helecho se cultivaron en macetas con un sustrato estéril y con un inóculo de G. intraradices (N° 51.331 BAFC) en la superficie. Después de la germinación, se tomaron muestras cada 15 días hasta que se completó el ciclo de vida del helecho. Con el fin de determinar la dinámica de la colonización, cada muestra se observó con el microscopio 864 óptico y el microscopio de confocal luego de la tinción correspondiente. El gametofito fue clasificado como del tipo “Adiantum”. Los gametangios femeninos y masculinos se desarrollaron en la cara inferior del mismo. La micorrización comenzó cuando los gametangios estaban ya diferenciados y la colonización se produjo a través de los rizoides. Las esporas y las vesículas no se encontraron en esta fase del ciclo. La micorrizacion tipo Paris se observó sobre la línea de la nervadura central. El pie del esporofito fue colonizado por el micelio externo. Cuando la raíz se desarrolló, se colonizó “de novo”, y se observó una colonización de tipo Arum. Este estudio demuestra que el tipo de colonización está determinado por la estructura del helecho y no por el hongo. Tanto el gametofito como el pie del embrión tienen crecimiento definido y colonización tipo Paris, mientras que las raíces del esporofito presentan un crecimiento indeterminado y una colonización tipo Arum. Las estructuras que se encontraron bajo cultivo coinciden con las que se encontraron en condiciones naturales. Ferns, which are usually colonizing different environments and their roots frequently present mycorrhization, have two adult stages in their life cycle, the sporophytic and the gametophytic phase. This paper describes the experimental mycorrhizal association between Pteris vittata leptosporangiate fern and a strain of Glomus intraradices during the life cycle of the fern, from spore germination to the development of a mature sporophyte. The aim of this study was to compare the colonization pattern of in vitro cultures of G. intraradices along the fern life cycle with those found in nature. For this, mature spores were obtained from fertile P. vittata fronds growing in walls of Buenos Aires city, Argentina. Roots were stained and observed under the light microscope for arbuscular mycorrhizal colonization. Approximately, 75 fern spores were cultured in each pot filled with a sterile substrate and G. intraradices (BAFC N° 51.331) as inoculum on the surface. After germination took place, samples were taken every 15 days until the fern cycle was completed. In order to determine colonization dynamics each sample was observed under optical and confocal microscope after staining. Gametophyte was classified as Adiantum type. Male and female gametangia were limited to the lower face, mycorrhizal colonization started when they were differentiated and took place through the rhizoids. Spores and vesicles were not found in this cycle stage. Paris-type mycorrhizal colonization was established in the midrib and in the embrionary foot. It was colonized by external mycelium. When the first root was developed soil inoculum colonized de novo this structure and Arum-type colonization was observed. This study proves that the type of colonization is determined by the structure of the host, not by the fungus. Both the gametophyte and embryo foot have determined growth and Paris-type colonization, while, sporophyte roots have undetermined growth and Arum-type colonization. The structures found in vitro cultures were highly similar to those found under natural conditions.

We examined the mycorrhizal type of 128 plant species in two patches of native vegetation of the Chaco Serrano Woodland, central Argentina, the largest dry forest area in South America. Of the 128 plant species investigated (belonging to 111 genera in 53 families), 114 were colonized by arbuscular mycorrhizal fungi (AM), orchid mycorrhizal associations were present in the five terrestrial orchid species analyzed, one ectomycorrhiza was only present in Salix humboldtiana Willd., and 96 harbored a dark septate endophyte (DSE) association. Co-occurrence of AM and DSE was observed in 88 plant species. We determine morphological types of arbuscular mycorrhizal fungi (Arum, Paris, and intermediate AM structures) and report the mycorrhizal status in 106 new species, 12 of which are endemic to central Argentina and two, Aa achalensis Schltr. and Buddleja cordobensis Griseb., are declared to be vulnerable species. Root colonization in the Chaco Serrano Woodland is widespread and should be considered in revegetation programs due to the deterioration of this particular ecosystem. Considering the predominance of AM and DSE associations and the various potential benefits that these associations may bring to plant establishment, they should receive special attention in conservation and reforestation of these woodlands.