Charles Pepe-Ranney

and 3 more

ABSTRACT Biological soil crusts (BSC) are key components of ecosystem productivity in arid lands and they cover a substantial fraction of the terrestrial surface. In particular, BSC N₂-fixation contributes significantly to the nitrogen (N) budget of arid land ecosystems. In mature crusts, N₂-fixation is largely attributed to heterocystous cyanobacteria, however, early successional crusts possess few N₂-fixing cyanobacteria and this suggests that microorganisms other than cyanobacteria mediate N₂-fixation during the critical early stages of BSC development. DNA stable isotope probing (DNA-SIP) with ¹⁵N₂ revealed that _Clostridiaceae_ and _Proteobacteria_ are the most common microorganisms that assimilate ¹⁵N₂ in early successional crusts. The _Clostridiaceae_ identified are divergent from previously characterized isolates, though N₂-fixation has previously been observed in this family. The Proteobacteria identified share >98.5 %SSU rRNA gene sequence identity with isolates from genera known to possess diazotrophs (e.g. _Pseudomonas_, _Klebsiella_, _Shigella_, and _Ideonella_). The low abundance of these heterotrophic diazotrophs in BSC may explain why they have not been characterized previously. Diazotrophs play a critical role in BSC formation and characterization of these organisms represents a crucial step towards understanding how anthropogenic change will affect the formation and ecological function of BSC in arid ecosystems. KEYWORDS: microbial ecology / stable isotope probing / nitrogen fixation / biological soil crusts