Mounds formed by pocket gophers (Geomys pinetis) in a longleaf pine—wiregrass savanna had larger daily soil temperature fluctuations than surrounding matrix locations. Microsite temperature differences persisted for more than a year, but were largest during summer months. Mound soil also had lower total carbon, total nitrogen, available ammonium, available phosphate and slightly lower soil moisture than matrix locations. These differences occurred in the context of an ecosystem where frequent fire consumes organic matter and removes herbaceous cover. The distinct microclimate and nutrient characteristics of gopher mounds did not significantly influence wiregrass germination, wiregrass seedling survival or longleaf pine seedling survival. Wiregrass germination and seedling survival in the first growing season was low and did not differ between mound and matrix locations. Longleaf pine seedling survival in the first growing season was 64–77% in the absence of fire, but no more than 15% in blocks that were burned. To see a relationship between mound microsite conditions and biotic response, it may be necessary to focus on microbes or turn to other ecosystems.