This dataset contains environmental and microbial diversity data relating to biological soil crusts collected at four locations in the Chihuahuan Desert. This study aims to 1) assess the carbon fixation capacity of biocrust types under varying incubation times; 2) assess biocrust nitrogen fixation capacity; 3) understand microbial community composition as well as biomass differences within these biocrust types; and 4) identify how geographic location and soil properties can explain differences seen in carbon fixation and microbial diversity. In order to characterize the ecology and microbial diversity ofbiocruststypes at a range of Chihuahuan Desert sites, vegetation and soil data was collected along with soil crust samples at 4 sites in southern New Mexico, USA: the Jornada Experimental Range (near T-West), the A-Mountain BLM site, the Volcanic BLM site, and White Sands Missile Range. Data and sample collection occurred between May and July of 2020, and the soil crust types of interest were light cyanobacterial, dark cyanobacterial, Peltula lichen, Clavascidium lichen, and moss crusts (n=5 samples for each type). After collection, crust samples were sequenced, targeting the 16S rRNA gene region for bacteria. This dataset includes the environmental context data, including vegetation cover (measured by the line-point intercept method) and soil chemistry data, as well as mean relative abundance for cyanobacterial species and phospholipid fatty acid analyses (PLFA) of crust samples for the purpose of comparing biomass, biodiversity and functional type differences between different biocrust types at the four sites. Soil metagenomic data is hosted at the NCBI Sequence Read Archive, and links to those data are provided in an inventory file. This study is complete.