This special issue reflects the recent advances in the field of environmental genomics and exposes the attractive prospects in the light of the new, rapidly-evolving tools that are next generation sequencing (NGS) approaches. Understanding the ecology, evolution, adaptation and biodiversity of organisms in their ecosystems is one of the most challenging scientific issues to which NGS and other “omics” may contribute. The papers deal with a broad range of organisms (eukaryotes and prokaryotes) and environments, including biotic interactions and methodological enhancements. The special issue highlights the exciting paths opened by NGS in environmental genomics and the novel opportunities to go deeper in the understanding and characterization of complex biological systems.