Phylogenetic resolution of deep eukaryotic and fungal relationships using highly conserved low-copy nuclear genes

A comprehensive and reliable eukaryotic tree of life is important for many aspects of biological studies from comparative developmental and physiological analyses to translational medicine and agriculture. Both gene-rich and taxon-rich approaches are effective strategies to improve phylogenetic accuracy and are greatly facilitated by marker genes that are universally distributed, well conserved, and orthologous among divergent eukaryotes. In this article, we report the identification of 943 low-copy eukaryotic genes andwe showthatmany of these genes arepromising tools in resolving eukaryotic phylogenies, despite the challenges of determining deep eukaryotic relationships. As a case study,we demonstrate that smaller subsets of ~20 and 52 genes could resolve controversial relationships among widely divergent taxa and provide strong support for deep relationships such as themonophyly and branching order of several eukaryotic supergroups. In addition, the use of these genes resulted in fungal phylogenies that are congruent with previous phylogenomic studies that usedmuch larger datasets, and successfully resolved several difficult relationships (e.g., forming a highly supported clade with Microsporidia, Mitosporidium and Rozella sister to other fungi). We propose that these genes are excellent for both gene-rich and taxon-rich analyses and can be applied atmultiple taxonomic levels and facilitate a more complete understanding of the eukaryotic tree of life.