Four myriapod relatives - but who are sisters? No end to debates on relationships among the four major myriapod subgroups.

BACKGROUND: Phylogenetic relationships among the myriapod subgroups Chilopoda, Diplopoda, Symphyla and Pauropoda are still not robustly resolved. The first phylogenomic study covering all subgroups resolved phylogenetic relationships congruently to morphological evidence but is in conflict with most previously published phylogenetic trees based on diverse molecular data. Outgroup choice and long-branch attraction effects were stated as possible explanations for these incongruencies. In this study, we addressed these issues by extending the myriapod and outgroup taxon sampling using transcriptome data. RESULTS: We generated new transcriptome data of 42 panarthropod species, including all four myriapod subgroups and additional outgroup taxa. Our taxon sampling was complemented by published transcriptome and genome data resulting in a supermatrix covering 59 species. We compiled two data sets, the first with a full coverage of genes per species (292 single-copy protein-coding genes), the second with a less stringent coverage (988 genes). We inferred phylogenetic relationships among myriapods using different data types, tree inference, and quartet computation approaches. Our results unambiguously support monophyletic Mandibulata and Myriapoda. Our analyses clearly showed that there is strong signal for a single unrooted topology, but a sensitivity of the position of the internal root on the choice of outgroups. However, we observe strong evidence for a clade Pauropoda+Symphyla, as well as for a clade Chilopoda+Diplopoda. CONCLUSIONS: Our best quartet topology is incongruent with current morphological phylogenies which were supported in another phylogenomic study. AU tests and quartet mapping reject the quartet topology congruent to trees inferred with morphological characters. Moreover, quartet mapping shows that confounding signal present in the data set is sufficient to explain the weak signal for the quartet topology derived from morphological characters. Although outgroup choice affects results, our study could narrow possible trees to derivatives of a single quartet topology. For highly disputed relationships, we propose to apply a series of tests (AU and quartet mapping), since results of such tests allow to narrow down possible relationships and to rule out confounding signal.

Sperm accessory microtubules suggest the placement of Diplura as the sister-group of Insecta s.s.

Sperm ultrastructure and spermiogenesis of the dipluran Japygidae (Japyx solifugus, Metajapyx braueri and Occasjapyx japonicus) and Campodeidae (Campodea sp.) were studied with the aim of looking for potential characters for the reconstruction of the phylogenetic relationships of basal hexapods. Both Japygidae and Campodeidae share a common sperm axonemal model 9+9+2, provided with nine accessory microtubules. These microtubules, however, after their formation lose the usual position around the 9+2 and migrate between the two mitochondria. In Japygidae, four of these microtubules are very short and were observed beneath the nucleus after negative staining and serial sections. Accessory microtubules have 13 protofilaments in their tubular wall. Diplura have a sperm morphology which is very different from that of the remaining Entognatha (Protura+Collembola). On the basis of the present results, the presence of accessory microtubules suggests that Diplura are the sister-group of the Insecta s.s.. Moreover, Japygidae and Campodeidae differ with regards to the relative position of the sperm components, the former having the axoneme starting from beneath the nucleus (above which sits the short acrosome), while the latter having a long apical acrosome and a nucleus running parallel with the proximal part of the axoneme. The present study also allowed to redescribe the male genital system of Japyx.