Chloroplast morphology and pyrenoid ultrastructural analyses reappraise the diversity of the lichen phycobiont genus Trebouxia (Chlorophyta)

Trebouxiophyceae is a wide class of green algae comprising coccoid and elliptic unicells, filaments, blades and colony-forming species that occur in diverse terrestrial and aquatic environments. Within this class, the genus Trebouxia Puymaly is among the most widespread lichen phycobionts worldwide. However, the 29 formally described species based on the combination of morphological traits and genetic diversity woefully underrepresented the overall species-level diversity recognized in the genus. In Trebouxia, reliable differentiation and characterization of the species-level lineages can be achieved by studying the diversity of key diagnostic features of pyrenoid ultrastructure and chloroplast morphology of axenically grown algal cultures. Here, we used transmission electron microscopy (TEM) coupled with confocal laser scanning microscopy (CLSM) to analyze the pyrenoid and the chloroplast of 20 Trebouxia species-level lineages grown directly on solid agar medium and on cellulose-acetate discs laid over the agar medium. With the new, detailed morphoanatomical characterization of these species-level lineages, we reappraise Trebouxia taxonomy in light of the most recent phylogenetic delimitation provided by Muggia et al. (2020).

Automated summary

Trebouxiophyceae is a diverse class of green algae found in terrestrial and aquatic environments. Among them, the genus Trebouxia is widely distributed in lichens worldwide. However, the current species descriptions do not fully represent the overall diversity of the genus. By studying the ultrastructure of pyrenoids and the morphology of chloroplasts, it is possible to differentiate and characterize species-level lineages effectively.