Identity and origins of introduced and native Azolla species in Florida

•Azolla pinnata, an exotic aquatic fern, is now in the northern Everglades.•Molecular/morphological tools indicate three A. pinnata subspecies.•These tools ID the exotic as A. pinnata subsp. pinnata, the Australian subspecies.•These tools ID the threatened native as Azolla caroliniana, a hybrid.•An unique Ecuadorian Azolla is the paternal ancestor of Azolla caroliniana.

Azolla pinnata, an introduced aquatic fern, is spreading rapidly causing concern that it may displace native Azolla. It is now present in the Arthur R. Marshall Loxahatchee National Wildlife Refuge, the northernmost portion of the Florida Everglades. Because A. pinnata subspecies are native to Africa, Southeast Asia, and Australia, determining the actual geographic origin of the Florida exotic is important to the discovery of efficacious biological control agents. Both the exotic and native Azollas were examined using morphological and molecular criteria. Both criteria distinguished three A. pinnata subspecies with the Florida exotic matching the Australian A. pinnata subsp. pinnata. Molecular divergence between the A. pinnata subspecies indicates the three types should be considered separate species. The Florida native was characterized by both molecular and morphological methods as Azolla caroliniana. The discovery of a previously uncharacterized Ecuadorian Azolla, which appears to be a paternal ancestor of A. caroliniana, indicates that A. caroliniana is a hybrid species.