Aytoniaceae are one of the largest families of complex thalloid liverworts (Marchantiopsida), consisting of about 70 species, with most species being distributed in temperate areas. However, the phylogeny and evolution of the morphological character of Aytoniaceae are still poorly understood. Here, we employed two chloroplast loci, specifically, rbcL and trnL-F, along with a 26S nuclear ribosomal sequence to reconstruct the phylogeny and track the morphological evolution of Aytoniaceae. Our results reveal that Aytoniaceae are monophyletic, and five monophyletic clades were recovered (i.e., Asterellopsis-Cryptomitrium, Calasterella, Mannia, Reboulia-Plagiochasma, and Asterella). Asterella was divided into five clades (i.e., Asterella lindenbergiana, subg. Saccatae, subg. Phragmoblepharis, subg. Wallichianae, and subg. Asterella), except for Asterella palmeri, which is the sister of Asterellopsis grollei. Bayesian molecular clock dating indicates that the five primary clades within Aytoniaceae underwent divergence events in the Cretaceous period. Asterellopsis differentiated during the early Upper Cretaceous (c. 84.2 Ma), and Calasterella originated from the late Lower Cretaceous (c. 143.0 Ma). The ancestral Aytoniaceae plant is reconstructed as the absence of a pseudoperianth, lacking equatorial apertures, and having both male and female reproductive organs on the main thallus. At present, Asterellopsis consists of two species known in Asia and America with the new transfer of Asterella palmeri to Asterellopsis. A new subgenus, Asterella subg. Lindenbergianae, is proposed.
Keywords: Asterella; Asterellopsis; ancestral character reconstructions; complex thalloid liverworts; divergence time; new combination.