Conservation scientists have adapted conservation planning principles designed for protection of habi- tats ranging from terrestrial to freshwater ecosystems. We applied current approaches in conservation planning to prioritize California watersheds for management of biodiversity. For all watersheds, we compiled data on the pre- sence/absence of herpetofauna and fishes; observations of freshwater-dependent mammals, selected invertebrates, and plants; maps of freshwater habitat types; measures of habitat condition and vulnerability; and current manage- ment status. We analyzed species-distribution data to identify areas of high freshwater conservation value that op- timized representation of target taxa on the landscape and leveraged existing protected areas. The resulting priority network encompasses 34% of the area of California and includes ≥10% of the geographic range for all target taxa. High-value watersheds supported nontarget freshwater taxa and habitats, and focusing on target taxa may provide broad conservation value. Most of the priority conservation network occurs on public lands (69% by area), and 46% overlaps with protected areas already managed for biodiversity. A significant proportion of the network area is on private land and underscores the value of programs that incentivize landowners to manage freshwater species and habitats. The priority conservation areas encompass more freshwater habitats/ha than existing protected areas. Land use (agriculture and urbanization), altered fire regimes, nonnative fish communities, and flow impairment are the most important threats to freshwater habitat in the priority network, whereas factors associated with changing climate are the key drivers of habitat vulnerability. Our study is a guide to a comprehensive approach to freshwater conservation currently lacking in California. Conservation resources are often limited, so prioritiza- tion tools are valuable assets to land and water managers.