Palynological and geochemical analyses of sediment cores collected on two tree islands in the Florida Everglades indicate long-term hydrologic and chemical differences between tree islands and surrounding marshes and sloughs. Gumbo Limbo and Nuthouse tree islands are elongate, teardrop-shaped islands in Water Conservation Area 3B. Prior to tree island formation at both sites, pollen records indicate that sites on modern tree island heads were covered with sawgrass marshes with abundant weedy annuals. Such vegetation is characteristic of moderate water depths and hydroperiods with frequent droughts or disturbances. Contemporaneously deposited sediments on tree island tails indicate progressively deeper water conditions with increasing distance from the head; wetlands surrounding tree islands were covered by sloughs with deep water and long hydroperiods. Tree island formation occurred at about 1200 BC on Gumbo Limbo Island, with mature tree island vegetation established by about 800 AD. On Nuthouse Island, tree island formation occurred around 300 AD, shifting to mature tree island vegetation around 1400 AD. Thus, tree island formation began on these islands between 3.2 Ka and 1.7 Ka. Maturation of tree islands took between 1,000 and 2,000 years, and vegetation on these tree islands has been relatively stable for the last 600–1,200 years. Phosphorus levels on tree island heads have been extremely high (approximately six times greater than baseline levels in marshes) throughout the history of the sites, and phosphorus content in tree island tails began increasing when tree island formation occurred. Elevated phosphorus content may reflect the long-term presence of wading birds at these sites and provide a proxy for reconstructing the historic distribution of wading bird populations.