The fossil flora of the Matzitzi Formation, is a fluvial unit of late Paleozoic age exposed in southern Mexico, has been studied since the last century; however, the discovery of new plant elements during the last decade imposes a reevaluation of two aspects that have always been the subject of debate: the depositional age of the unit and the taxonomy of its paleoflora. The objective of the present work is to report new paleofloral records from two areas of the Matzitzi Formation as well as their chronological implications. The study sites are named Carretera and Coatepec. Nineteen genera and 11 species have been identified in beds from the Carretera site, whereas 23 genera and 16 species in beds from the Coatepec site. The analysis of floral assemblages of both sites shows that they share 49% of taxa. Ovopteris, Lesleya, Lobatopteris, and Trigonocarpus are unique to the Carretera site, while Coatepec is characterized by the presence of cf. Bjuvia, Schizoneura, and Velascophyllum. When analyzing the greatest ranges of the paleoflora awareness for the Carretera site indicates that deposition of these beds took place in a time interval that may span between Pennsylvanian and lower Permian. On the other side, the floral assemblage recovered from the Coatepec site indicates that the hosting beds were deposited in upper Permian time. U–Pb detrital zircon geochronology indicates that the maximum depositional age of the fossiliferous beds at the Carretera and Coatepec sites are of 276 ± 15 Ma and 254 ± 11 Ma, respectively. The integration of the paleontological and geochronological data obtained in this work suggests that the analyzed floral assemblage recovered from the Carretera site is of Kungurian age, whereas the one from the Coatepec site is of Lopingian age. Alternatively, we explore the possibility that deposits that are presently grouped within the Matzitzi Formation could be the vestiges of different fluvial cycles associated with different tectonic settings. Finally, the work on this unit has not finished, since there are still localities where its flora and age are still unknown.