Evolutionary mobility and genetic dynamics of MORFFO genes: shuttling among ancient plant lineages

Plastid genomes (plastomes) of land plants are characterized by their architectural and genic content stability. However, fern plastomes exhibit unexpected dynamism, characterized by the presence of mobile protein-coding genes (CDS) - Mobile Open Reading Frames in Fern Organelles (MORFFOs). We investigate the evolutionary dynamics of MORFFOs in 30 species of Anemiaceae (Schizaeales), an ancient lineage of ferns, focusing on their transposition, substitution patterns, codon usages, and RNA editing patterns. MORFFOs expand plastome size and occur in diverse intergenic regions, exhibiting dynamic locations, genealogies, and exceptionally high substitution rates compared with canonical plastid CDS. Sliding window and codon usage analyses demonstrate that MORFFOs are under purifying selection but exhibit distinct codon preferences that deviate from those of other plastid CDS, suggesting functional constraints. Phylogenetic incongruence between MORFFOs and other plastid CDS, along with their extraordinary substitution rates and mobility, implies their replication outside plastids. Our findings highlight that MORFFOs are dynamic, potentially selfish genetic elements capable of transcription, translation, and replication independently from plastomes, and fern plastomes might acquire these mobile CDS through frequent horizontal gene transfer and possibly intracellular gene transfer.