A phylogenetic analysis of the genus Dichotrachelus (Curculionidae: Cyclominae) was carried out, based on a morphological matrix and, for some species, on mitochondrial cytochrome oxidase I sequences. Bayesian Inference, Maximum Likelihood and Maximum Parsimony were implemented and the results were compared. The genus is found to be isolated in the subfamily, not related to the only other Palaearctic tribe (Hipporhinini) and possibly nearer to the south-American genera of Cyclominae of the tribe Listroderini. Among these, Macrostyphlus is also equally associated to mosses as the host plant. In Dichotrachelus, two main clades were recognized, one distributed in the western part of the Mediterranean region (Iberian Peninsula, northern Africa and southern France) and the second distributed in the Alps and Apennines. Within each clade, some differentiated monophyletic subgroups could be identified. An evaluation of the most important characters that led to the phylogenetic reconstruction indicated the male genital sclerite as the most useful structure to characterize the different clades. updated in 2015 [20]. Some additional species of the genus were also included in a more generic Barcode-approach on Swiss Alpine weevils [21].Herein, we propose a phylogenetic analysis for the entire genus Dichotrachelus, based on a morphological database matrix, as well as on mt-Cox1 sequences available for some species, in order to support the main clades (or species groups) and recognize the synapomorphies that define these groups. Available sequences of other Cyclominae were used in a first attempt at recognizing the affinities of Dichotrachelus within Cyclominae.
Material and Methods
Samples OriginSee Table I in the supplementary materials for a list of the species and specimens used, and for authorship and year of publication of the names. Specimens for molecular analyses were sampled by the authors. The weevils were hand-collected, immediately killed in 95% ethanol and preserved at −20 • C. Attempts to recover complete sequences of other species from dry collection specimens were seldom successful, and usually did not provide any reliable data. For the morphological analysis, samples from Meregalli's collection were used. Almost all species were included in the morphological matrix, except for D. elongatus and D. ulbrichi, species only known from the types, or, for the former, from very few specimens which could not be analysed, and the descriptions of which are insufficiently detailed to permit the scoring of the character states used for the study. Different subspecies of the same species were united with the nominal species, with the exception of D. knechti. This species was classified as a subspecies of D. stierlini in [1], but in the morphological analysis some of the character-states were distinct from those of D. stierlini, hence we have included it in the study. Sequences of the other species of Cyclominae were retrieved from GenBank.
Outgroup SelectionThe selection of an outgroup proved particularly complex, since, a...