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ABSTRACT Investigation of microbial communities has led to many advances in our understanding of ecosystem function, whether that ecosystem is a subglacial lake or the human gut. Within these communities, much emphasis has been placed on interspecific variation and between-species relationships. However, with current advances in sequencing technology resulting in both the reduction in sequencing costs and the rise of shotgun metagenomic sequencing, the importance of intraspecific variation and within-species relationships is becoming realized. Our group conducts multi-omic analyses to understand how spatial structure and resource availability influence diversification within a species and the potential for long-term coexistence of multiple ecotypes within a microbial community. Here, we present examples of ecotypic variation observed in the lab and in the wild, current challenges faced when investigating intraspecies diversity, and future developments that we expect to define the field over the next 5 years.