A nested polymerase chain reaction (PCR) assay was used to detect phytoplasmas in stem tissues from declining silver date (Phoenix sylvestris), Canary Island date (Phoenix canariensis), edible date (Phoenix dactylifera), Queen (Syagrus romanozoffiana) and Mexican fan (Washingtonia robusta) palms, all displaying symptoms similar to lethal yellowing (LY) disease, in a tri-county region of west central Florida. Restriction fragment length polymorphism analysis or sequencing of PCR-amplified rDNA products (1.6 kb) identified three distinct group 16SrIV phytoplasma strains among 38 infected palms. Most palms (86.8%) contained Texas Phoenix decline (TPD) phytoplasma, a subgroup 16SrIV-D strain, while two P. canariensis harboured palm LY phytoplasma, a subgroup 16SrIV-A strain. A novel phytoplasma, initially detected in W. robusta and tentatively classified as a subgroup 16SrIV-F strain, also occurred with LY phytoplasma as mixed infections in two P. dactylifera palms. Collectively, these findings extend the known geographic range of TPD in the USA to include Florida and the listing of TPD-susceptible palm species to include P. dactylifera, P. sylvestris and S. romanozoffiana. Moreover, discovery of a novel subgroup 16SrIV-F strain in W. robusta adds to the list of phytoplasma host palm species and complexity of phytoplasma-vector-palm pathosystems newly identified in the west central region of the state. The emergence of new lethal disease of palms beyond southern subtropical region of the state is unprecedented but follows similar developments in other regions where LY is known to occur.