Prototaxites was an extinct lineage of multicellular terrestrial eukaryotes

Prototaxites was the first giant organism to live on the terrestrial surface, reaching sizes of 8 metres in the Early Devonian. However, its taxonomic assignment has been debated for over 165 years1-7. Tentative assignments to groups of multicellular algae or land plants1,2,8-11 have been repeatedly ruled out based on anatomy and chemistry5,7,11-16 resulting in two major alternatives: Prototaxites was either a fungus5,6,17-22 or a now entirely extinct lineage 7,16,23. Recent studies have converged on a fungal affinity5-7,17-20,22. Here we test this by contrasting the anatomy and molecular composition of Prototaxites with contemporary fungi from the 407-million-year-old Rhynie chert. We report that Prototaxites taiti was the largest organism in the Rhynie ecosystem and its anatomy was fundamentally distinct from all known extant or extinct fungi. Furthermore, our molecular composition analysis indicates that cell walls of P. taiti include aliphatic, aromatic, and phenolic components most similar to fossilisation products of lignin, but no fossilisation products characteristic of chitin or chitosan, which are diagnostic of all groups of extant and extinct fungi, including those preserved in the Rhynie chert. We therefore conclude that Prototaxites was not a fungus, and instead propose it is best assigned to a now entirely extinct terrestrial lineage.