Two new subspecies belonging to the genus Globorotalia, that we name G. mariae mariae and G. mariae gratiae, are here described and figured. They were found exclusively within the Middle Miocene and particularly from the early Langhian to the Serravallian of several Mediterranean sedimentary successions. The two subspecies have a slightly different stratigraphic range. Both were recorded from the Globigerinoides sicanus/Orbulina suturalis Zone (MMi4) to the Paragloborotalia partimlabiata Zone (MMi7). However, only G. mariae mariae is recorded from the Globorotalia praescitula Subzone (MMi4b), whereas G. mariae gratiae was found from slightly younger levels, corresponding to the Paragloborotalia siakensis-Praeorbulina glomerosa circularis Subzone (MMi4c). As far as their origin concerns, G. praescitula may have been the ancestor of G. mariae mariae. Subsequently, G. mariae gratiae evolved from G. mariae mariae. Both subspecies disappear in the Paragloborotalia partimlabiata-Globoturborotalita druryi Subzone (MMi7a). Concerning their stratigraphic range it is important to point out that in all the studied sections both subspecies were not found in the interval encompassing the Langhian/Serravallian boundary, and lasting from the upper part of the Praeorbulina glomerosa circularis Subzone (MMi4d) to the uppermost part of the Orbulina suturalis/Globorotalia peripheroronda Zone (MMi5).