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
INTRODUCTORY PARAGRAPH We explored the microbial contributions to decomposition using a sophisticated approach to DNA Stable Isotope Probing (SIP). Our experiment evaluated the dynamics and ecological characteristics of functionally defined microbial groups that metabolize labile and structural C in soils. We added to soil a complex amendment representing plant derived organic matter substituted with either ¹³C-xylose or ¹³C-cellulose to represent labile and structural C pools derived from abundant components of plant biomass. We found evidence for ¹³C-incorporation into DNA from ¹³C-xylose and ¹³C-cellulose in 49 and 63 operational taxonomic units (OTUs), respectively. The types of microorganisms that assimilated ¹³C in the ¹³C-xylose treatment changed over time being predominantly _Firmicutes_ at day 1 followed by _Bacteroidetes_ at day 3 and then _Actinobacteria_ at day 7. These ¹³C-labeling dynamics suggest labile C traveled through different trophic levels. In contrast, microorganisms generally metabolized cellulose-C after 14 days and did not change to the same extent in phylogenetic composition over time. Microorganisms that metabolized cellulose-C belonged to poorly characterized but cosmopolitan soil lineages including _Verrucomicrobia_, _Chloroflexi_ and _Planctomycetes_.