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Abstract Antarctic soils are unique from those found nearly anywhere else on Earth yet can still harbor a broad diversity of microorganisms able to tolerate the challenging conditions typical of the continent. For these reasons, microbiologists have been drawn to Antarctica for decades. However, our understanding of which microbes thrive in Antarctic soils and how they to do so remains limited. To help resolve these knowledge gaps, we analyzed a collection of 200 archived Antarctic soils—from Livingston Island on the Antarctic Peninsula to Cape Hallett in northern Victoria Land. We analyzed the prokaryotic and fungal communities in these soils using both cultivation-independent marker gene sequencing and cultivation-dependent approaches (microbial isolation), paired with extensive soil geochemical analyses. Our cultivation-independent analyses indicate that colder, saltier, and drier soils harbor less diverse communities of bacteria and fungi, distinct from those found in soils with less challenging conditions. We also built a culture collection from a subset of these soils that encompasses more than 50 bacterial and fungal genera, including cold-tolerant organisms, such asCryobacteriumandCryomyces. By directly comparing the diversity of our cultured isolates against our cultivation-independent data, we show that many of the more abundant Antarctic taxa are not readily cultivated and highlight bacterial and fungal taxa that should be the focus of future cultivation efforts. Together, we hope that our collection of isolates, the comprehensive data compiled from the cultivation-independent analyses, and our geochemical analyses will serve as a community resource to accelerate the study of Antarctic soil microbes. 

This data package offers comprehensive insights into Antarctic soil microbial diversity and composition. From 2003 to 2023, a total of 186 samples were collected from diverse locations spanning the Antarctic Peninsula to East Antarctica, representing a wide range of environmental gradients and climatic conditions. Soils were stored at -20°C to preserve their integrity for downstream analyses. This data package integrates cultivation-independent sequencing of prokaryotic and fungal communities alongside a robust cultivation-dependent culture collection to enable direct comparisons across microbial diversity assessment methods. Accompanying geochemical, physicochemical, and environmental parameters provide critical context for biogeographical analyses, offering a valuable resource for studying microbial adaptations and community dynamics in extreme Antarctic environments.