Isolation of biocrust cyanobacteria and evaluation of Cu, Pb, and Zn immobilisation potential for soil restoration and sustainable agriculture.

Soil contamination by heavy metals represents an important environmental and public health problem of global concern. Biocrust-forming cyanobacteria offer promise for heavy metal immobilisation in contaminated soils due to their unique characteristics, including their ability to grow in contaminated soils and produce exopolysaccharides (EPS). However, limited research has analysed the representativeness of cyanobacteria in metal-contaminated soils. Additionally, there is a lack of studies examining how cyanobacteria adaptation to specific environments can impact their metal-binding capacity. To address this research gap, we conducted a study analysing the bacterial communities of cyanobacteria-dominated biocrusts in a contaminated area from South Sardinia (Italy). Additionally, by using two distinct approaches, we isolated three Nostoc commune strains from cyanobacteria-dominated biocrust and we also evaluated their potential to immobilise heavy metals. The first isolation method involved acclimatizing biocrust samples in liquid medium while, in the second method, biocrust samples were directly seeded onto agar plates. The microbial community analysis revealed Cyanobacteria, Bacteroidota, Proteobacteria, and Actinobacteria as the predominant groups, with cyanobacteria representing between 13.3 % and 26.0 % of the total community. Despite belonging to the same species, these strains exhibited different growth rates (1.1-2.2 g L-1 of biomass) and capacities for EPS production (400-1786 mg L-1). The three strains demonstrated a notable ability for metal immobilisation, removing up to 88.9 % of Cu, 86.2 % of Pb, and 45.3 % of Zn from liquid medium. Cyanobacteria EPS production showed a strong correlation with the removal of Cu, indicating its role in facilitating metal immobilisation. Furthermore, differences in Pb immobilisation (40-86.2 %) suggest possible environmental adaptation mechanisms of the strains. This study highlights the promising application of N. commune strains for metal immobilisation in soils, offering a potential bioremediation tool to combat the adverse effects of soil contamination and promote environmental sustainability.

Biocrusts intensify water redistribution and improve water availability to dryland vegetation: insights from a spatially-explicit ecohydrological model.

Biocrusts are ecosystem engineers in drylands and structure the landscape through their ecohydrological effects. They regulate soil infiltration and evaporation but also surface water redistribution, providing important resources for vascular vegetation. Spatially-explicit ecohydrological models are useful tools to explore such ecohydrological mechanisms, but biocrusts have rarely been included in them. We contribute to closing this gap and assess how biocrusts shape spatio-temporal water fluxes and availability in a dryland landscape and how landscape hydrology is affected by climate-change induced shifts in the biocrust community. We extended the spatially-explicit, process-based ecohydrological dryland model EcoHyD by a biocrust layer which modifies water in- and outputs from the soil and affects surface runoff. The model was parameterized for a dryland hillslope in South-East Spain using field and literature data. We assessed the effect of biocrusts on landscape-scale soil moisture distribution, plant-available water and the hydrological processes behind it. To quantify the biocrust effects, we ran the model with and without biocrusts for a wet and dry year. Finally, we compared the effect of incipient and well-developed cyanobacteria- and lichen biocrusts on surface hydrology to evaluate possible paths forward if biocrust communities change due to climate change. Our model reproduced the runoff source-sink patterns typical of the landscape. The spatial differentiation of soil moisture in deeper layers matched the observed distribution of vascular vegetation. Biocrusts in the model led to higher water availability overall and in vegetated areas of the landscape and that this positive effect in part also held for a dry year. Compared to bare soil and incipient biocrusts, well-developed biocrusts protected the soil from evaporation thus preserving soil moisture despite lower infiltration while at the same time redistributing water toward downhill vegetation. Biocrust cover is vital for water redistribution and plant-available water but potential changes of biocrust composition and cover can reduce their ability of being a water source and sustaining dryland vegetation. The process-based model used in this study is a promising tool to further quantify and assess long-term scenarios of climate change and how it affects ecohydrological feedbacks that shape and stabilize dryland landscapes.

Towards large scale biocrust restoration: Producing an efficient and low-cost inoculum of N-fixing cyanobacteria.

Dryland soil degradation is increasing due to global change and traditional restoration methods are not successful due to water scarcity. Thus, an alternative technology based on inoculating biocrust-forming cyanobacteria on degraded soils has emerged. Biocrusts are communities of mosses, lichens, cyanobacteria or fungi that colonize soil surface forming a stable and fertile layer. Previous studies have shown the benefits of inoculating cyanobacteria to restore soils at a small scale. However, to face field restoration projects, it is necessary to produce high quantities of biomass at an affordable cost. In this work, we analyze if the previously tested cyanobacteria Scytonema hyalinum, Tolypothrix distorta (heterocystous strains) and Trichocoleus desertorum (a bundle-forming one) can be produced with agricultural fertilizers. Different culture media were used: two containing pure chemicals (BG11 and BG110, this N-free medium was used just for heterocystous strains) and two containing fertilizers (BG11-F and MM-F). The performance of the cultures was monitored by measuring the biomass concentration and photosynthetic stress. Afterwards, we analyzed their capacity to induce biocrusts and improve soil properties by inoculating the biomass on a mine substrate indoors and measuring, three months later, the albedo, chlorophyll a and organic carbon content. Results show that the bundle-forming cyanobacterium was unable to grow in the media tested, whereas both heterocystous cyanobacteria grew in all of them and induced the formation of biocrusts improving the organic carbon substrate content. The best results for S. hyalinum were found using the MM-F medium, and for T. distorta using a medium containing pure chemicals (BG11). However, results were also positive when using a medium containing fertilizers (BG11-F). Thus, agricultural fertilizers can be used to undertake the production of heterocystous cyanobacteria for large scale restoration in drylands. On the other hand, more research is needed to find sustainable techniques to produce biomass of bundle-forming cyanobacteria.

Non-rainfall water inputs: A key water source for biocrust carbon fixation.

Links between water and carbon (C) cycles in drylands are strongly regulated by biocrusts. These widespread communities in the intershrub spaces of drylands are able to use non-rainfall water inputs (NRWI) (fog, dewfall and water vapour) to become active and fix carbon dioxide (CO2), converting biocrusts into the main soil C contributors during periods in which vegetation remains inactive. In this study, we first evaluated the influence of biocrust type on NRWI uptake using automated microlysimeters, and second, we performed an outdoor experiment to examine how NRWI affected C exchange (photosynthesis and respiration) in biocrusts. NRWI uptake increased from incipient cyanobacteria to well-developed cyanobacteria and lichen biocrusts. NRWI triggered biocrust activity but with contrasting effects on CO2 fluxes depending on the main NRWI source. Fog mainly stimulated respiration of biocrust-covered soils, reaching net CO2 emissions of 0.68 µmol m-2 s-1, while dew had a greater effect stimulating biocrust photosynthesis and resulted in net CO2 uptake of 0.66 µmol m-2 s-1. These findings demonstrate the key role that NRWI play in biocrust activity and the soil C balance in drylands.

Polyphasic evaluation of key cyanobacteria in biocrusts from the most arid region in Europe.

Cyanobacteria are key microbes in topsoil communities that have important roles in preventing soil erosion, carbon and nitrogen fixation, and influencing soil hydrology. However, little is known regarding the identity and distribution of the microbial components in the photosynthetic assemblages that form a cohesive biological soil crust (biocrust) in drylands of Europe. In this study, we investigated the cyanobacterial species colonizing biocrusts in three representative dryland ecosystems from the most arid region in Europe (SE Spain) that are characterized by different soil conditions. Isolated cyanobacterial cultures were identified by a polyphasic approach, including 16S rRNA gene sequencing, phylogenetic relationship determination, and morphological and ecological habitat assessments. Three well-differentiated groups were identified: heterocystous-cyanobacteria (Nostoc commune, Nostoc calcicola, Tolypothrix distorta and Scytonema hyalinum), which play an important role in N and C cycling in soil; nonheterocystous bundle-forming cyanobacteria (Microcoleus steenstrupii, Trichocoleus desertorum, and Schizothrix cf. calcicola); and narrow filamentous cyanobacteria (Leptolyngbya frigida and Oculatella kazantipica), all of which are essential genera for initial biocrust formation. The results of this study contribute to our understanding of cyanobacterial species composition in biocrusts from important and understudied European habitats, such as the Mediterranean Basin, a hotspot of biodiversity, where these species are keystone pioneer organisms.

Ecology and functional roles of biological soil crusts in semi-arid ecosystems of Spain.

Biological soil crusts (BSCs), composed of lichens, cyanobacteria, mosses, liverworts and microorganisms, are key biotic components of arid and semi-arid ecosystems worldwide. Despite they are widespread in Spain, these organisms have been historically understudied in this country. This trend is beginning to change as a recent wave of research has been identifying BSCs as a model ecological system. Many studies and research projects carried out in Spain have explored the role of BSCs on water, carbon and nitrogen fluxes, the interactions between BSCs and vascular plants, their dynamics after disturbances, and their response to global change, among other topics. In this article we review the growing body of research on BSCs available from semi-arid areas of Spain, highlighting its importance for increasing our knowledge on this group of organisms. We also discuss how it is breaking new ground in emerging research areas on the ecology of BSCs, and how it can be use to guide management and restoration efforts. Finally, we provide directions for future research on the ecology of BSCs in Spain and abroad.