Assessing the validity of the two-subfamily system in the Nemouridae: An integrated taxonomic approach.

Phylogenomic approaches have recently helped to elucidate evolutionary relationships across numerous taxa. Nemouridae (Order Plecoptera) is a Holarctic family divided into two subfamilies, Amphinemurinae and Nemourinae, with seven and 14 extant genera, respectively, plus an additional three genera that are currently known only from the fossil record. The separation of the two subfamilies has been predicated, in part, on the presence of three paraproct lobes on adult males (Amphinemurinae) vs. two (Nemourinae). However, the validity of the two subfamily system in Nemouridae has yet to be holistically addressed using more modern phylogenetic methods, in part, due to limited taxon sampling across the family. In this study, we sequenced the mitochondrial genome of 13 Nemouridae species from China, among which 2 genera (Illiesonemoura, Zapada) were sequenced for the first time. In addition, we sequenced low coverage whole genomes of 10 Nemouridae species from China, including 4 genera (Illiesonemoura, Indonemoura, Sphaeronemoura, Mesonemoura) for the first time. The site-heterogeneous model C20+F+R was applied to a maximum likelihood analysis of Nemouridae for the first time. All conclusions support the monophyly of the Amphinemurinae, but the Nemourinae is not monophyletic. All results confirmed Lednia + (Prostoia + Podmosta) as the sister group to the remaining genera of Nemouridae.