SignificanceCyanobacteria were responsible for the origin of oxygenic photosynthesis, and have since come to colonize almost every environment on Earth. Here we show that their ecological range is not limited by the presence of sunlight, but also extends down to the deep terrestrial biosphere. We report the presence of microbial communities dominated by cyanobacteria in the continental subsurface using microscopy, metagenomics, and antibody microarrays. These cyanobacteria were related to surface rock-dwelling lineages known for their high tolerance to environmental and nutritional stress. We discuss how these adaptations allow cyanobacteria to thrive in the dark underground, a lifestyle that might trace back to their nonphotosynthetic ancestors.

Deep continental subsurface is defined as oligotrophic environments where microorganisms present a very low metabolic rate. To date, due to the energetic cost of production and maintenance of biofilms, their existence has not been considered in poor porous subsurface rocks. We applied fluorescence in situ hybridization techniques and confocal laser scanning microscopy in samples from a continental deep drilling project to analyze the prokaryotic diversity and distribution and the possible existence of biofilms. Our results show the existence of natural microbial biofilms at all checked depths of the Iberian Pyrite Belt (IPB) subsurface and the co-occurrence of bacteria and archaea in this environment. This observation suggests that multi-species biofilms may be a common and widespread lifestyle in subsurface environments.

Río Tinto (Huelva, southwestern Spain) is an extreme environment with a remarkably constant acidic pH and a high concentration of heavy metals, conditions generated by the metabolic activity of chemolithotrophic microorganisms thriving in the rich complex sulfides of the Iberian Pyrite Belt (IPB). Fungal strains isolated from the Tinto basin were characterized morphologically and phylogenetically. The strain identified as Purpureocillium lilacinum specifically induced the formation of a yellow-ocher precipitate, identified as hydronium-jarosite, an iron sulfate mineral which appears in abundance on the banks of Río Tinto. The biomineral was characterized by X-ray diffraction (XRD) and its formation was observed with high-resolution transmission electron microscopy (TEM) and scanning electron microscopy (SEM) coupled to energy-dispersive X-ray spectroscopy (EDX) microanalysis. Jarosite began to nucleate on the fungal cell wall, associated to the EPS, due to a local increase in the Fe(3+) /Fe(2+) ratio which generated supersaturation. Its formation has been also observed in non-viable cells, although with much less efficiency. The occurrence of P. lilacinum in an ecosystem with high concentrations of ferric iron and sulfates such as Río Tinto suggests that it could participate in the process of jarosite precipitation, helping to shape and control the geochemical properties of this environment.