Endosymbioses are a major form of biological complexity affecting the ecological and evolutionary diversification of many eukaryotic groups. These associations are exemplified by nutritional symbioses of insects for which phylogenetic studies have demonstrated numerous cases of long-term codiversification between a bacterial and a host lineage. Some insects, including most leafhoppers (Insecta: Hemiptera: Cicadellidae), have more than one bacterial symbiont within specialized host cells, raising questions regarding the patterns of codiversification of these multiple partners and the evolutionary persistence of complex symbiotic systems. Previous studies reported the presence of two dominant symbiont types in a member of the leafhopper subfamily Cicadellinae (sharpshooters). In this study, 16S rRNA sequences were obtained and used to examine the occurrence and evolutionary relationships of the two dominant symbiont types across 29 leafhopper species. Candidatus Sulcia muelleri (Bacteroidetes) was detected in all leafhopper species examined, a finding that is consistent with a previous report of its ancient association with the Auchenorrhyncha (a grouping that includes leafhoppers, treehoppers, cicadas, planthoppers, and spittlebugs). Baumannia cicadellinicola (Proteobacteria), previously known from only five sharpshooter species, was found only in the sharpshooter tribes Cicadellini and Proconiini, as well as in the subfamily Phereurhininae. Mitochondrial and nuclear gene sequences were obtained and used to reconstruct host phylogenies. Analyses of host and symbiont data sets support a congruent evolutionary history between sharpshooters, Sulcia and Baumannia and thus provide the first strong evidence for long-term co-inheritance of multiple symbionts during the diversification of a eukaryotic host. Sulcia shows a fivefold lower rate of 16S rDNA sequence divergence than does Baumannia for the same host pairs. The term 'coprimary' symbiont is proposed for such cases.

A data set comprising DNA sequences from 388 loci and >99,000 aligned nucleotide positions, generated using anchored hybrid enrichment, was used to estimate relationships among 138 leafhoppers and treehoppers representative of all major lineages of Membracoidea, the most diverse superfamily of hemipteran insects. Phylogenetic analysis of the concatenated nucleotide sequence data set using maximum likelihood produced a tree with most branches receiving high support. A separate coalescent gene tree analysis of the same data generally recovered the same strongly supported clades but was less well resolved overall. Several nodes pertaining to relationships among leafhopper subfamilies currently recognized based on morphological criteria were separated by short internodes and received low support. Although various higher taxa were corroborated with improved branch support, relationships among some major lineages of Membracoidea are only somewhat more resolved than previously published phylogenies based on single gene regions or morphology. In agreement with previous studies, the present results indicate that leafhoppers (Cicadellidae) are paraphyletic with respect to the three recognized families of treehoppers (Aetalionidae, Melizoderidae, and Membracidae). Divergence time estimates indicate that most of the poorly resolved divergence events among major leafhopper lineages occurred during the lower to middle Cretaceous and that most modern leafhopper subfamilies, as well as the lineage comprising the three recognized families of treehoppers, also arose during the Cretaceous.

This is the author manuscript accepted for publication and has undergone full peer review but has not been through the copyediting, typesetting, pagination and proofreading process, which may lead to differences between this version and the Version of Record. Please cite this article as

Hydroptilidae constitute the most diverse caddisfly family, with over 2000 species known from every habitable continent. Leucotrichiinae are exclusively New World microcaddisflies, currently including over 200 species and 17 genera. Phylogenetic analyses of Leucotrichiinae relationships based on 114 morphological characters and 2451 molecular characters from DNA sequences were conducted. DNA sequences analysed were from one mitochondrial gene, cytochrome oxidase I (653 bp), and four nuclear genes, carbamoylphosphate synthase (802 bp), elongation factor 1 (352 bp), histone 3 (308 bp) and 28S rDNA (336 bp). The morphological matrix included 94 taxa (with representatives of all included genera) and the molecular matrix included 62 taxa. Individual and combined datasets were analysed under parsimony and Bayesian inference. In addition, a relaxed molecular clock divergence time estimate was conducted to determine the age of the subfamily and major lineages. All Bayesian inference analyses strongly suggest a monophyletic Leucotrichiinae, which initially diverged at approximately 124 Ma into two monophyletic groups of genera. These groups are herein elevated to tribal status, Alisotrichiini trib.n. and Leucotrichiini Flint sensu n. Several genera of Leucotrichiini were not recovered as monophyletic clades and some nomenclatural changes are proposed to reflect their phylogenetic history. These include the synonymy of Abtrichia with Peltopsyche; transfer of Betrichia hamulifera to Costatrichia; transfer of Betrichia alibrachia and Costatrichia falsa to Leucotrichia; and transfer of Costatrichia fluminensis to Acostatrichia. Additionally, Tupiniquintrichia gen.n. is described to include Peltopsyche maclachlani and Leucotrichia procera. According to our results, crown diversifications of both Alisotrichiini trib.n. (∼80 Ma) and Leucotrichiini sensu n. (∼103 Ma) occurred after complete separation of South America from Africa. Current distributions of most leucotrichiine genera are probably a result of migration from South America towards the north using the proto-Caribbean archipelago (100 to 49 Ma).This published work has been registered in ZooBank, http://zoobank.org/urn:lsid: zoobank.org:pub:FB6A3385

"Triatoma brasiliensis species complex" was defined as a monophyletic group of the species: T. brasiliensis, T. juazeirensis, T. melanica, and T. sherlocki. An alternative grouping scheme proposed the concept of "Brasiliensis subcomplex" which included the former species together with T. melanocephala, T. petrocchiae, T. lenti, T. tibiamaculata, and T. vitticeps. To evaluate the relationship among these taxa we combined the results obtained with four mitochondrial genes (12S, 16S, COI and Cytb, adding to 1811bp) and geometric morphometric analysis of wings and heads. Panstrongylus megistus was included in the analysis as it was previously found related to T. tibiamaculata, T. melanocephala and T. vitticeps. The results of both molecular and morphometric approaches clearly grouped the species analyzed into two monophyletic units, supported by both genetic and wing variability. The first one (G1) comprises the four species originally included in the T. brasiliensis species complex plus T. lenti and T. petrocchiae. The second group (G2) was composed by T. melanocephala, T. tibiamaculata and T. vitticeps, and remarkably, P. megistus if considering wing variability and phylogenetic results. Nevertheless, geometric morphometrics of heads provided a quantitative measurement that discriminates Panstrongylus from the Triatoma species based on the position of the antennal insertion relative to eyes, as it is used as the generic distinctive character. The discrepancy among approaches questions the validity of this character to define Panstrongylus genus. Independently of the chosen group definition -"T. brasiliensis species complex" or "Brasiliensis subcomplex"-we propose to delimit it to species of G1 that are all associated with the Caatinga biome in the Brazilian Northeast. G2 are the ones associated with the Atlantic Forest biome.

Multiple fragments of mitochondrial DNA genes (cytochrome b, cytochrome oxidase I, and 16S rDNA) were used to evaluate the phylogenetic relationships among Triatoma melanocephala, Triatoma tibiamaculata, Triatoma vitticeps, and other members of Triatoma brasiliensis subcomplex under a Bayesian framework and maximum parsimony criterion. With the addition of new sequences of T. tibiamaculata and T. vitticeps, Triatoma juazeirensis, Triatoma melanica and the newly sequenced T. melanocephala, the three first sylvatic species, T. melanocephala, T. tibiamaculata and T. vitticeps, were strongly recovered into a clade separate from the other with the remaining Triatoma species from South America, such as the members of T. brasiliensis subcomplex. Panstrongylus megistus was recovered as a sister to T. tibiamaculata, whereas T. vitticeps was a sister to T. melanocephala. This study revealed the non-monophyly of the T. brasiliensis subcomplex, and the polyphyly of Triatoma was reinforced by the placement of these three sylvatic species with Dipetalogaster, Meccus, Mepraia, and Panstrongylus. The results herein shown highlight the need of generic revision in Triatomini.

The spittlebug family Cercopidae is currently divided into two subfamilies: the paraphyletic Old World Cercopinae and the monophyletic New World Ischnorhininae. The most recent classification scheme proposed by Fennah in 1968 divided the New World Cercopidae into four tribes: Tomaspidini, Ischnorhinini, Hyboscartini and Neaenini. Herein we present a phylogenetic analysis of Ischnorhininae using 108 morphological characters and including 53 of the 59 recognized genera, to evaluate the tribal-level classification and understand the processes underlying the current distributional patterns of these genera. We found significant support for the monophyly of many Neotropical genera, but Fennah's tribal classification is revised because tribes Neaenini, Ischnorhinini and Tomaspidini were recovered as polyphyletic. Hyboscartini was synonymized with Tomaspidini. A taxonomic key to tribes and genera of Neotropical spittlebugs is provided based mostly on recovered apomorphies. The biogeographical analysis suggests a Neotropical origin of ischnorhinines, more specifically in northwestern South America. This was possibly coincident spatially and temporally with the origin of grasses, with ancestral range expansions southward to the Amazonian and Paraná regions, and posterior vicariant events, possibly related to the expansion of forests in the Chacoan region, the South America diagonal of open formations. Dispersals to the Chacoan region and to the Nearctic region are hypothesized to have occurred only within genera. In the Chacoan region it is associated with more recent events, such as the diversification of C4 grasses and establishment of the savannas and seasonally dry forests.

No abstract