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CandidatusNanohaloarchaeota, an archaeal phylum within the DPANN superphylum, is characterized by limited metabolic capabilities and limited phylogenetic diversity and until recently has been considered to exclusively inhabit hypersaline environments due to an obligate association withHalobacteria. Aside from hypersaline environments,Ca.Nanohaloarchaeota can also have been discovered from deep-subsurface marine sediments.

Three metagenome-assembled genomes (MAGs) representing a new order within theCa.Nanohaloarchaeota were reconstructed from a stratified salt crust and proposed to represent a novel order,Nucleotidisoterales. Genomic features reveal them to be anaerobes capable of catabolizing nucleotides by coupling nucleotide salvage pathways with lower glycolysis to yield free energy. Comparative genomics demonstrated that these and otherCa.Nanohaloarchaeota inhabiting saline habitats use a “salt-in” strategy to maintain osmotic pressure based on the high proportion of acidic amino acids. In contrast, previously describedCa.Nanohaloarchaeota MAGs from geothermal environments were enriched with basic amino acids to counter heat stress. Evolutionary history reconstruction revealed that functional differentiation of energy conservation strategies drove diversification withinCa.Nanohaloarchaeota, further leading to shifts in the catabolic strategy from nucleotide degradation within deeper lineages to polysaccharide degradation within shallow lineages.

This study provides deeper insight into the ecological functions and evolution of the expanded phylumCa.Nanohaloarchaeota and further advances our understanding on the functional and genetic associations between potential symbionts and hosts.