High spatial heterogeneity and low connectivity of bacterial communities along a Mediterranean subterranean estuary

20 pages, 8 figures, 2 tables, supporting Information https://doi.org/10.1111/mec.16695.-- Data availabitiy statement: DNA sequences and associated metadata: European Nucleotide Archive (ENA) under accession numbers PRJEB52186. The environmental metadata and the nonrarefied ASV and taxonomic tables are provided as Tables S2–S4
Subterranean estuaries are biogeochemically active coastal sites resulting from the underground mixing of fresh aquifer groundwater and seawater. In these systems, microbial activity can largely transform the chemical elements that may reach the sea through submarine groundwater discharge (SGD), but little is known about the microorganisms thriving in these land-sea transition zones. We present the first spatially-resolved characterization of the bacterial assemblages along a coastal aquifer in the NW Mediterranean, considering the entire subsurface salinity gradient. Combining bulk heterotrophic activity measurements, flow cytometry, microscopy and 16S rRNA gene sequencing we find large variations in prokaryotic abundances, cell size, activity and diversity at both the horizontal and vertical scales that reflect the pronounced physicochemical gradients. The parts of the transect most influenced by freshwater were characterized by smaller cells and lower prokaryotic abundances and heterotrophic production, but some activity hotspots were found at deep low-oxygen saline groundwater sites enriched in nitrite and ammonium. Diverse, heterogeneous and highly endemic communities dominated by Proteobacteria, Patescibacteria, Desulfobacterota and Bacteroidota were observed throughout the aquifer, pointing to clearly differentiated prokaryotic niches across these transition zones and little microbial connectivity between groundwater and Mediterranean seawater habitats. Finally, experimental manipulations unveiled large increases in community heterotrophic activity driven by fast growth of some rare and site-specific groundwater Proteobacteria. Our results indicate that prokaryotic communities within subterranean estuaries are highly heterogeneous in terms of biomass, activity and diversity, suggesting that their role in transforming nutrients will also vary spatially within these terrestrial–marine transition zones
This work was funded by the Spanish Ministry of Science, Innovation and Universities (MICINN) through the GRAMMI project (RTI2018-099740-J-I00), the Ramon y Cajal contract (RYC2019-026758-I) and partly by the project MEDISTRAES III (PID2019-110311RB-C22 and PID2019-110212RB-C21). Additional financial support was provided by grants SPIP2020-02595 (Ministry for the Ecological Transition and the Demographic Challenge [MITECO]) with funding from the Spanish government through the ‘Severo Ochoa Centre of Excellence’ accreditation (CEX2019-000928-S). JGO acknowledges the financial support of the Spanish Ministry of Science, Innovation and Universities, through the “Maria de Maeztu” programme for Units of Excellence (CEX2019-000940-M). VR acknowledges financial support from the Beatriu de Pinós postdoctoral programme of the Catalan Government (2019-BP-00241). AAK acknowledges financial support from ICTA “Unit of Excellence” (MinECo, MDM2015-0552-17-1) and PhD fellowship, BES-2017-080740. The coauthor AF is a Serra Húnter Fellow. MDF acknowledges the economic support from the FI-2017 fellowships of the Generalitat de Catalunya autonomous government (2017FI_B_00365). The authors would like to thank the support of the Generalitat de Catalunya to MERS (2017 SGR-1588), GHS (2017 SGR 1485) and 2017SGR/156, for additional fundin