Two-dimensional (2D) material powders are featured with large specific surface areas, ample catalytically active sites, high solution processibilities, and widely tunable microstructures, showing some tremendous application prospects in various fields, such as energies, electronics, catalysis, and environments, etc. Low-cost and batch production of high-quality 2D material powders with tunable microstructures is the vital prerequisite for developing their large-scale applications. This review summarized the research progresses on diatomite-templated syntheses of 2D material powders (e.g., graphene, graphidiyne, transition metal nitrides, and transition metal dichalcogenides powders), and introduced the advances in the applications of as-prepared 2D material powders in energy-storage devices, printed electronics, electrocatalytic hydrogen evolution reactions, and wastewater treatments, etc. In addition, some problems and challenges in diatomite-templated syntheses of 2D material powders were represented, and their potential application directions were also discussed.