Literature on distributions, habitat characteristics, and identification of Opuntia species in the southeastern US is scant. Regional treatments of Opuntia are often ambiguous, limited studies based primarily on herbarium specimens. The present work provides a detailed description of the opuntias found in Mississippi along with their distributions and habitat descriptions. Observations, collections, and measurements were made from natural populations within Mississippi and other parts of the southeastern United States. Plants of all taxa were greenhouse grown for approximately two years to monitor morphological characteristics, which sometimes can be quite plastic. Our two-year study encompassed nearly 150 natural populations among several eastern states. Five species are recognized. Of these, Opuntia cespitosa, O. humifusa, and O. pusilla are the most common in Mississippi. O. aff allairei and O. stricta are found infrequently.