Drastic mycorrhizal community shifts in Sceptridium ferns during the generation transition from fully mycoheterotrophic gametophytes to photosynthetic sporophytes.

Many plant species experience a prolonged subterranean phase during which they rely entirely on mycorrhizal fungi for carbon. While this mycoheterotrophic strategy spans liverworts, lycophytes, and ferns, most empirical research has centered on angiosperms. This study explores the fungal associations of Sceptridium (Ophioglossaceae), an early-diverging fern with mycoheterotrophic gametophytes. We analyzed germination patterns and fungal associations in Sceptridium gametophytes, comparing them to the distribution and mycorrhizal partners of photosynthetic sporophytes. High-throughput sequencing data reveal that mycoheterotrophic gametophytes consistently associate with a single Entrophospora fungus in the order Entrophosporales (Glomeromycotina), while photosynthetic sporophytes primarily partner with fungi from Glomeraceae (Glomerales, Glomeromycotina). Consequently, gametophytes exhibit spatial clustering without association with adult plants. This is the first documentation of an association between Entrophosporaceae (and the order Entrophosporales) and mycoheterotrophic plants. The drastic shifts in Sceptridium mycorrhizal communities across life stages likely reflect changing physiological needs during development. Further research is essential to determine whether the association with Entrophosporaceae is widespread among mycoheterotrophic species and to elucidate the functional and physiological mechanisms underlying these mycorrhizal shifts.