Macroecology of biodiversity disentangles local and regional drivers of biodiversity by exploring large-scale biodiversity relationships with environmental or biotic gradients, generalizing local biodiversity relationships across regions, or comparing biodiversity patterns among species groups. A macroecological perspective is also important at local scales: a full understanding of local biodiversity drivers, including human impact, demands that regional processes be taken into account. This requires knowledge of which species could inhabit a site (the species pool), including those that are currently absent (dark diversity). Macroecology of biodiversity is currently advancing quickly owing to an unprecedented accumulation of biodiversity data, new sampling techniques and analytical methods, all of which better equip us to face current and future challenges in ecology and biodiversity conservation.