Groundwater ecosystems constitute the largest terrestrial freshwater biome. They are dark, extremely low in energy and do not provide much space but they contain an unexpectedly high diversity of living forms showing characteristic adaptive features. The restricted accessibility along with the enormous 'invisible' heterogeneity challenged for a long time testing of scientific theories and unraveling of ecosystem functioning. Triggered by an improved interdisciplinarity, comprehensive sampling strategies and current developments in biotechnology and statistical analysis, groundwater ecology gains momentum entering a new era of research. We are only beginning to understand adaptive mechanisms, species distribution patterns and ecosystem functioning. Ninety-five percent of global liquid freshwater is stored in the terrestrial subsurface constituting a major source of water for drinking, irrigation and industrial purposes. There is an urgent need to integrate evolutionary and ecological research for developing a holistic perspective of the functional roles of biodiversity and ecosystem services and predicting global changes under alternative groundwater resource use scenarios.