Molecular phylogeny and systematics of the Pieridae (Lepidoptera: Papilionoidea): higher classification and biogeography

The systematic relationships of the butterfly family Pieridae are poorly understood. Much of our current understanding is based primarily on detailed morphological observations made 50–70 years ago. However, the family and its putative four subfamilies and two tribes, have rarely been subjected to rigorous phylogenetic analysis. Here we present results based on an analysis of molecular characters used to reconstruct the phylogeny of the Pieridae in order to infer higher‐level classification above the generic level and patterns of historical biogeography. Our sample contained 90 taxa representing 74 genera and six subgenera, or 89% of all genera recognized in the family. Three complementary approaches were employed: (1) a combined analysis of a 30 taxon subset for sequences from four gene regions, including elongation factor‐1 alpha (EF‐1α), wingless, cytochrome oxidase subunit I (COI), and 28S (3675 bp, 1031 parsimony‐informative characters), mainly to establish higher‐level relationships, (2) a single‐gene analysis of the 90 taxon data set for sequences from EF‐1α (1066 bp, 364 parsimony‐informative characters), mainly to establish lower‐level relationships, and (3) an all available data analysis of the entire data set for sequences from the four genes, to recover both deep and shallow nodes. Analyses using maximum parsimony, maximum likelihood and Bayesian inference provided similar results. All supported monophyly for the four subfamilies but not for the two tribes, with the Anthocharidini polyphyletic and the Pierini paraphyletic. The combined and all available data analyses support the following relationships among the subfamilies: ((Pseudopontiinae + Dismorphiinae) + (Coliadinae + Pierinae)), corroborating Ehrlich’s 1958 phenetic hypothesis. On the basis of these analyses, and additional morphological and life history evidence, we propose a reclassification of the subfamily Pierinae into two tribes (Anthocharidini s.s., Pierini s.s.) and two informal groups (Colotis group, Leptosia), with the tribe Pierini s.s. subdivided into three subtribes (Appiadina, Pierina, Aporiina) and three genera (Elodina, Dixeia, Belenois) of uncertain status (incertae sedis). The combined and all available data analyses support the following relationships among the Pierinae: (Colotis group + Anthocharidini s.s. + Leptosia + (Elodina + ((Dixeia + Belenois) + Appiadina + Pierina + Aporiina))). Application of a molecular clock calibrated using fossil evidence and semiparametric rate smoothing suggests that divergence between the Pierina and Aporiina occurred no later than the Palaeocene (> 60 Myr). The minimum estimate for the age of the crown‐group of the Pieridae was 112–82 Myr, with a mean of 95 Myr. A historical biogeographical hypothesis is proposed to explain the present‐day distribution of the clade Pseudopontiinae + Dismorphiinae, which argues for an origin of the two subfamilies in western Gondwana (Africa + South America) during the Late Cretaceous. © 2006 The Linnean Society of London, Zoological Journal of the Linnean Society, 2006, 147 , 239–275.     

Here be dragons: a phylogenetic and biogeographical study of the Smaug warreni species complex (Squamata: Cordylidae) in southern Africa

Taxonomy of the Smaug warreni species complex remains contentious despite known morphological differences and geographical separation of the various taxa. This study uses an 11‐gene dataset to recover phylogenetic relationships between the seven nominal members of the S. warreni complex. Eight well‐supported clades were returned, with S. warreni barbertonensis found to be paraphyletic. A time‐calibrated analysis of molecular data indicates that all eight clades in the S. warreni complex separated in the late Miocene, much earlier than the date suggested by the existing hypothesis of vicariance through the ingression of Kalahari sands. Ecological niche modelling indicates that although all clades are allopatric, a slight decrease in temperature could potentially render them sympatric, supporting an hypothesis of range expansion through climatic change. © 2014 The Linnean Society of London     

Monophyletic groups within 'higher land birds'– comparison of morphological and molecular data

The relationships within the ‘higher land birds’ and putatively related taxa are analysed in a study using 89 morphological characters and DNA sequences of three nuclear, protein‐coding genes, c‐myc, RAG‐1, and myoglobin intron II. Separate analyses of the different data sets and a ‘total evidence’ analysis in which the data sets of the morphological and molecular analyses were combined are compared. All three analyses support the hitherto disputed sister group relationship between Pici (Ramphastidae, Indicatoridae and Picidae) and Galbulae (Galbulidae and Bucconidae). Previously unrecognized osteological synapomorphies of this clade are presented. All analyses further resulted in monophyly of the taxon [Aegothelidae + (Apodidae/Hemiprocnidae + Trochilidae)]. Analysis of the morphological data and of the combined data set also supported monophyly of the taxon [Strigiformes + (Falconidae + Accipitridae)]. The morphological data further support monophyly of the taxon (Upupidae + Bucerotidae). Other placements in the three analyses received either no or only weak bootstrap support.     

How many marmoset (Primates: Cebidae: Callitrichinae) genera are there? A phylogenetic analysis based on multiple morphological systems

The marmosets, tribe Callitrichini, are the most speciose clade in the subfamily Callitrichinae, containing 21 species. However, there is no consensus among molecular and morphological systematists as to how many genera should be recognized for the group. To test the morphological support for the alternative generic classifications, this study presents a comprehensive phylogenetic analysis. It is the first such analysis to include all 21 species and employ continuous and discrete osteological, pelage and tegument, karyological and vocal characters. This dataset was combined with nucleotide sequences from two mitochondrial and four nuclear regions. Separate analyses showed that, among morphological datasets, osteological characters were best at solving relationships at more inclusive levels, whilst pelage characters were most informative at the interspecific level. This suggests the presence of different transformation rates for the two character sets. When a single most parsimonious tree was obtained using the 83‐character matrix, three main clades were identified, supporting the division of the marmosets into three genera: Callithrix, Cebuella and Mico. The total evidence analysis that included an additional 3481 molecular characters corroborated most of the morphology‐based clades and also supported a three‐genus classification of the marmosets. This is the first morphological study to support an Amazonian marmoset clade (Cebuella + Mico), which is also strongly supported in exclusively molecular phylogenies, and to synonimize Callibella under Mico.     

Phylogeny of the Neuropterida: a first molecular approach

Abstract. In a first molecular approach specially dedicated to examining the phylogeny of the Neuropterida, two nuclear and two mitochondrial genes were tested: 18S rRNA, translation elongation factor‐1α, cytochrome c oxidase subunit 3 and 16S rRNA. Molecular results are discussed in the light of a previous holomorphological cladistic analysis. The hypothesis of a sister‐group relationship Raphidioptera + (Neuroptera + Megaloptera) put forward in recent morphological analyses is supported by our data, which is in contrast to the traditional view (Raphidioptera + Megaloptera) + Neuroptera. Furthermore, the Nevrorthidae (constituting the suborder Nevrorthiformia) as a sister group of all other Neuroptera is confirmed. The disruption of the suborder Hemerobiiformia is the most conflicting result of the molecular analysis. Sisyridae and Osmylidae do not cluster within Hemerobiiformia, but represent two distinct and widely separated branches. The remaining Hemerobiiformia emerge as the sister group of the suborder Myrmeleontiformia, which is once more confirmed as monophyletic. Among the genes tested, cytochrome c oxidase subunit 3 proved to be most potent for resolving the phylogenetic relationships among Neuropterida. The nuclear gene for the ribosomal 18S rRNA is too conserved within the alignable regions, whereas the variable sections are too divergent to be applicable within this evolutionary time frame. The elongation factor‐1α gene proved to exist in more than one copy in Neuropterida, and thus is not applicable in the present state of knowledge. With respect to the mitochondrial sequences (cytochrome c oxidase subunit 3, 16S rRNA), saturation impedes the unambiguous resolution of deeper nodes. Apparently, due to early diversification of the heterogeneous Neuroptera, phylogenetic analysis of this group remains a challenge with respect to selection of the proper genes and mutatis mutandis the morphological approach.     

Mosaic nature in the skeleton of East Asian crocodylians fills the morphological gap between “Tomistominae” and Gavialinae

Crocodylian systematics has long been confounded by conflicting hypotheses of higher level relationships—although molecular data sets strongly supported the sister-taxon relationship of Tomistoma and Gavialis, morphological data sets placed Gavialis as sister to all other living taxa. One of the perceived difficulties in interpreting morphological character evolution on the molecular tree is the extensive character reversal occurring in Gavialinae, the mechanism of which has yet to be explained. Here, we provide evidence of gavialine-specific atavistic characters from East Asian “tomistomines” Penghusuchus pani and Toyotamaphimeia machikanensis. These taxa exhibit a mosaic assembly of “tomistomine” and gavialine features, which fill the gap between the two longirostrine groups. Although the parsimony analysis of morphological data (69 taxa, 254 characters) still supports the previous morphological hypothesis, the alternative tree that was forced to fit the molecular hypothesis was insignificantly (5/954 steps; 0.52%) longer than the unconstrained tree, suggesting that morphological evolution can also be interpreted on the molecular tree. Although the problem of stratigraphic gaps remains, future studies may be directed to resolving the interrelationships within Gavialoidea, a large longirostrine group of crocodylians, in the molecular tree context.     

Phylogenetic position of the house dust mite subfamily Guatemalichinae (Acariformes: Pyroglyphidae) based on integrated molecular and morphological analyses and different measures of support

Based on multilocus phylogenetic analyses (18S, 28S, EF1‐α, SRP54, HSP70, CO1, 10 860 nt aligned), we show that the house dust mite subfamily Guatemalichinae is nested within non‐onychalgine pyroglyphid mites and forms the sister group to the genus Sturnophagoides (bootstrap support 100, posterior probability 1.0). Because high bootstrap support values may be misleading in the presence of incongruence, we evaluate robustness of the Guatemalichinae+Sturnophagoides clade using: (1) internode certainty indices to estimate the frequency of conflicting bipartitions in maximum‐likelihood bootstrap trees, (ii) consensus networks to investigate conflict among different loci; and (iii) statistical hypothesis testing based on information theory, both multi‐scale and regular bootstrap. Results suggest that this grouping is very well supported given the data. The molecular analyses were integrated with detailed morphological study using scanning electron and light microscopy. We suggest that the subfamilial status of Guatemalichinae should be reconsidered, and this lineage should be placed within the subfamily Dermatophagoidinae. The latter subfamily is currently accepted in the literature as a monophyletic group but was here inferred as paraphyletic and was not supported by any morphological synapomorphy. The paraphyly involved the most species‐rich and medically important genus, Dermatophagoides. Our findings suggest the need for a comprehensive revision of the higher‐level relationships of pyroglyphid house dust mites using both DNA sequences and morphology coupled with a broad taxonomic sampling.     

Phylogenetic relationships of flesh flies in the subfamily Sarcophaginae based on three mtDNA fragments (Diptera: Sarcophagidae)

In an effort to improve our knowledge of the phylogenetic relationships among species and genera of the subfamily Sarcophaginae, we analysed data from three mitochondrial gene fragments. Sequence data for portions of the genes cytochrome oxidase I (COI), cytochrome oxidase II (COII) and dehydrogenase subunit 4 (ND4) were obtained from 43 species of Sarcophagidae representing 15 genera. We used a Bayesian approach to simultaneously choose how best to partition the data and which substitution model to apply to each partition. Phylogenetic relationships were inferred using Bayesian Inference and Maximum Likelihood methods. Our results are consistent with monophyly of the subfamily Sarcophaginae (posterior probability 1; bootstrap support 93%), as well as with monophyly of several genera within the Sarcophaginae (including Sarcophaga s.l.; posterior probability 1; bootstrap support 97%). We found support for a sister‐group relationship between Ravinia Robineau‐Desvoidy and Oxysarcodexia Townsend, which has been hypothesised by past authors on the basis of morphological similarities, although this was supported only in the Bayesian analyses (posterior probability 0. 81–0. 98), and for some novel supra‐generic clades. Contrary to a recent morphological hypothesis, we do not find Helicobia Coquillett to be nested within Sarcophaga Meigen; our data suggest, but do not strongly support, a hypothesis that Peckia Robineau‐Desvoidy is the sister group to Sarcophaga.     

The deep divergences of neornithine birds: a phylogenetic analysis of morphological characters

Consensus is elusive regarding the phylogenetic relationships among neornithine (crown clade) birds. The ongoing debate over their deep divergences is despite recent increases in available molecular sequence data and the publication of several larger morphological data sets. In the present study, the phylogenetic relationships among 43 neornithine higher taxa are addressed using a data set of 148 osteological and soft tissue characters, which is one of the largest to date. The Mesozoic non‐neornithine birds Apsaravis, Hesperornis, and Ichthyornis are used as outgroup taxa for this analysis. Thus, for the first time, a broad array of morphological characters (including both cranial and postcranial characters) are analyzed for an ingroup densely sampling Neornithes, with crown clade outgroups used to polarize these characters. The strict consensus cladogram of two most parsimonious trees resultant from 1000 replicate heuristic searches (random stepwise addition, tree‐bisection‐reconnection) recovered several previously identified clades; the at‐one‐time contentious clades Galloanseres (waterfowl, fowl, and allies) and Palaeognathae were supported. Most notably, our analysis recovered monophyly of Neoaves, i.e., all neognathous birds to the exclusion of the Galloanseres, although this clade was weakly supported. The recently proposed sister taxon relationship between Steatornithidae (oilbird) and Trogonidae (trogons) was recovered. The traditional taxon “Falconiformes” (Cathartidae, Sagittariidae, Accipitridae, and Falconidae) was not found to be monophyletic, as Strigiformes (owls) are placed as the sister taxon of (Falconidae + Accipitridae). Monophyly of the traditional “Gruiformes” (cranes and allies) and ”Ciconiiformes” (storks and allies) was also not recovered. The primary analysis resulted in support for a sister group relationship between Gaviidae (loons) and Podicipedidae (grebes)—foot‐propelled diving birds that share many features of the pelvis and hind limb. Exclusion of Gaviidae and reanalysis of the data set, however, recovered the sister group relationship between Phoenicopteridae (flamingos) and grebes recently proposed from molecular sequence data.     

Molecular systematics and evolution of the Ptinidae (Coleoptera: Bostrichoidea) and related families

The Ptinidae (Coleoptera: Bostrichoidea) are a cosmopolitan, ecologically diverse, but poorly known group of Coleoptera and, excluding a few economic pests, species are rarely encountered. This first broad phylogenetic study of the Ptinidae s.l. (i.e. including both the spider beetles and anobiids) examines relationships based on DNA sequence data from two mitochondrial genes (16S and COI) and one nuclear gene (28S), using out‐group taxa from both the Bostrichidae and Dermestidae. Topologies varied depending on the genes used and whether data were analysed with either parsimony or Bayesian methods. Generally the two mitochondrial genes supported relationships near the tips of the phylogeny, whereas the nuclear gene supported the basal relationships. The monophyly of the Ptinidae was not inferred by all of the gene combinations and analysis methods, although the combined Ptinidae and Bostrichidae have a single origin in all cases. Alternative relationships include the Ptinidae s.s. (i.e. Ptininae and Gibbiinae) as sister to the anobiids (i.e. the nine remaining subfamilies of Ptinidae s.l.) + Bostrichidae, or the Bostrichidae as sister to the Ptinidae s.s.+ anobiids. Most of the larger subfamilies within the Ptinidae are not monophyletic. Further analysis with more taxa and more genes will be required to clarify and decide upon the best hypothesis of relationships found within the clades of the Bostrichidae and Ptinidae. © 2012 The Linnean Society of London, Zoological Journal of the Linnean Society, 2012, 165 , 88–108.     

Relationships within Podocarpaceae based on DNA sequence,anatomical, morphological,and biogeographical data

Despite considerable recent progress in understanding intergeneric relationships, a comprehensive analysis of Podocarpaceae at the species level using molecular data, biogeography, anatomy, and morphology has not been previously attempted. Here we present sequence analyses of rbcL, nrITS1 and NEEDLY intron 2 for two‐thirds (183 accessions of 145 taxa) of all Podocarpaceae species representing all genera except Parasitaxus. These analyses include many more species and accessions than previous studies and result in a more resolved phylogeny. The comprehensive anatomical and morphological study ensures that the identification of taxa is correct and also provides clade support. Bayesian and parsimony analyses were used to resolve 20 well‐supported monophyletic groups including 11 groups of the formerly poorly resolved subgenera Podocarpus and Foliolatus. The well‐resolved topology is supported by anatomical and morphological features and is highly congruent with geographical distribution. © The Willi Hennig Society 2011.     

Phylogenetic relationships of Steinernema Travassos, 1927 (Nematoda: Cephalobina: Steinernematidae) based on nuclear, mitochondrial and morphological data

Entomopathogenic nematodes of the genus Steinernema are lethal parasites of insects that are used as biological control agents of several lepidopteran, dipteran and coleopteran pests. Phylogenetic relationships among 25 Steinernema species were estimated using nucleotide sequences from three genes and 22 morphological characters. Parsimony analysis of 28S (LSU) sequences yielded a well-resolved phylogenetic hypothesis with reliable bootstrap support for 13 clades. Parsimony analysis of mitochondrial DNA sequences (12S rDNA and cox 1 genes) yielded phylogenetic trees with a lower consistency index than for LSU sequences, and with fewer reliably supported clades. Combined phylogenetic analysis of the 3-gene dataset by parsimony and Bayesian methods yielded well-resolved and highly similar trees. Bayesian posterior probabilities were high for most clades; bootstrap (parsimony) support was reliable for approximately half of the internal nodes. Parsimony analysis of the morphological dataset yielded a poorly resolved tree, whereas total evidence analysis (molecular plus morphological data) yielded a phylogenetic hypothesis consistent with, but less resolved than trees inferred from combined molecular data. Parsimony mapping of morphological characters on the 3-gene trees showed that most structural features of steinernematids are highly homoplastic. The distribution of nematode foraging strategies on these trees predicts that S. hermaphroditum, S. diaprepesi and S. longicaudum (US isolate) have cruise forager behaviours.     

Phylogeny and classification of whirligig beetles (Coleoptera: Gyrinidae): relaxed‐clock model outperforms parsimony and time‐free Bayesian analyses

Phylogenetic relationships among members of the family Gyrinidae (Coleoptera: Adephaga) were inferred from analysis of 42 morphological characters and DNA sequence data from the genes 12S rRNA, cytochrome c oxidase I and II, elongation factor 1 alpha (2 different copies) and histone III. Eighty‐nine species of Gyrinidae were included representing all known subfamilies, tribes and genera. Outgroups include species from Noteridae, Paelobiidae and Dytiscidae. Analyses include parsimony analysis, and partitioned time‐free and relaxed‐clock Bayesian analyses of the combined data using reversible‐jump MCMC to simultaneously integrate over all possible 4 × 4 nucleotide substitution models. Analyses resulted in conflicting topologies between the combined parsimony and Bayesian analyses on the one hand, and the relaxed‐clock analysis on the other. The marginal likelihoods of competing models were calculated with stepping‐stone sampling and used in a Bayes factor test, which, along with arguments from morphology, supported the topology generated by the relaxed‐clock analysis. This phylogenetic hypothesis is adopted to revise the higher classification of Gyrinidae. Major taxonomic conclusions include: (i) monophyletic Gyrinidae, (ii) the Nearctic Spanglerogyrinae Folkerts (with one species, Spanglerogyrus albiventris Folkerts) sister to all other Gyrinidae, (iii) the Madagascar endemic Heterogyrinae Brinck stat. n. (with one species, Heterogyrus milloti Legros) sister to all Gyrinidae except Spanglerogyrinae, (iv) monophyletic Gyrininae Latreille including three monophyletic tribes with the following relationship: Orectochilini Régimbart + (Gyrinini Latreille + Enhydrini Régimbart), (v) monophyletic Orectochilini comprising four monophyletic genera with the following relationships: (Gyretes Brullé + Patrus Aubé stat. n. ) + (Orectogyrus Régimbart + Orectochilus Dejean), (vi) monophyletic Gyrinini comprising three genera with the following relationships: Gyrinus Geoffroy + (Metagyrinus Brinck + Aulonogyrus Motschulsky), each monophyletic except Metagyrinus with only one included species and not tested for monophyly, and (vii) monophyletic Enhydrini comprising five genera with the following relationships: (Porrorhynchus Laporte + Dineutus MacLeay) + (Enhydrus Laporte + (Andogyrus Ochs + Macrogyrus Régimbart)), each monophyletic except Porrorhynchus, Enhydrus and Andogyrus each with one included species and untested for monophyly. Each subfamily, tribe and genus is diagnosed and discussed. The female reproductive tract of each group is presented, illustrated and discussed with respect to the phylogenetic conclusions.     

Phylogenetic analysis of the genus Hexachlamys (Myrtaceae) based on plastid and nuclear DNA sequences and their taxonomic implications

Myrtaceae are one of the most species‐rich families of flowering plants in the Neotropics. They include several complex genera and species; Hexachlamys is one of the complex genera. It has not been recognized as a distinct genus and has been included in Eugenia, based on morphological grounds. Therefore, molecular systematic studies may be useful to understand and to help to solve these relationships. Here, we performed a molecular phylogenetic analysis using plastid and nuclear data in order to check the inclusion of Hexachlamys in Eugenia. Plastid (accD, rpoB, rpoC1, trnH‐psbA) and nuclear (ITS2) sequence data were analysed using Bayesian and maximum parsimony methods. The trees constructed using ITS2 and trnH‐psbA were the best able to resolve the relationships between species and genera, revealing the non‐monophyly of Hexachlamys. The molecular phylogenetic analyses were in agreement with previous morphological revisions that have included Hexachlamys in Eugenia. These results reinforce the importance of uniting knowledge and strategies to understand better issues of delimitation of genera and species in groups of plants with taxonomic problems. © 2013 The Linnean Society of London, Botanical Journal of the Linnean Society, 2013, 172 , 532–543.     

Phylogenetic relationships of the family Axinellidae (Porifera: Demospongiae) using morphological and molecular data

Alvarez, B., Crisp, M.D., Driver, F., Hooper, J.N.A. & Van Soest, R.W.M. (2000). Phylogenetic relationships of the family Axinellidae (Porifera: Demospongiae) using morphological and molecular data. —Zoologica Scripta, 29, 169–198. Twenty‐seven species of marine sponges belonging to Axinellidae and related groups (Halichondriidae, Dictyonellidae, Agelasida) were selected to test the monophyly of Axinellidae and investigate their phylogenetic relationships using parsimony and maximum likelihood methods. Partial 28S rDNA sequences, including the D3 domain, and traditional morphological characters (mainly skeletal ones) were used independently to construct phylogenetic trees. Sequences were aligned using the appropriate model of secondary structure of the RNA and compared to that produced by the multiple sequence alignment program, ClustalW. The alignment using secondary structure constraints produced a better estimate of the phylogeny and was demonstrated to be an effective and objective method. Results of the cladistic analyses of the molecular and morphological data sets were not fully congruent; the morphological data suggest that Axinellidae is monophyletic, however, the molecular data suggest that it is nonmonophyletic. The single most‐parsimonious tree derived from the molecular data showed that species of Axinella (except A. polypoides) are united in a clade that is more closely related to members of Agelasida than to other species of Axinellidae; the remaining members of Axinellidae form a monophyletic group that is closely related to the families Dictyonellidae and Halichondriidae. The consensus tree of 20 most‐parsimonious trees from the morphological analysis, on the other hand, showed that all the sampled species of Axinellidae belong to a monophyletic group which is closely related to the species of Dictyonellidae and Halichondriidae. Only two branches were identical in both cladograms, the one uniting the species of Ptilocaulis and Reniochalina and the one with the species of Dictyonellidae. The robustness of the molecular and morphological trees (or parts of the trees), was tested using bootstrap, jack‐knife, PTP and T‐PTP tests. The results of the PTP test were significant indicating significant cladistic structure in both data sets. The bootstrap and jack‐knife values indicate that the molecular tree is in general better supported than the morphological one. The lack of morphological characters and the homoplastic nature of some may explain the weak support of the morphological tree. A T‐PTP test of nonmonophyly showed that the nonmonophyly of Axinellidae, as indicated by the results of the molecular analysis, is not significant; however, a T‐PTP test of monophyly of Axinellidae, as indicated by the morphological tree, produced significant results. This indicates that the monophyly of Axinellidae based on morphological data cannot be rejected; the family however, cannot be defined in terms of a unique diagnostic character common to all members of the ingroup. Tests of heterogeneity (reciprocal T‐PTP and partition homogeneity test) indicated that the data partitions are heterogeneous, which could be due to sampling errors (in either data set) or differences in the underlying phylogenies; therefore data were not combined in a single analysis. Further, both data sets are unequally sized (95 informative molecular characters vs. 16 informative morphological characters), which means that the molecular signal could swamp the morphological signal if the data is combined. Nonmonophyly of Axinellidae is supported by chemical and genetic evidence available in the literature and DNA sequences data of axinellid species from New Zealand. However, this needs to be confirmed using independent evidence from different genes (or gene regions), biochemistry, histology or cell ultrastructure. Therefore, no changes to the taxonomic position of the family in the higher classification are proposed at this stage.     

Phylotranscriptomics resolves ancient divergences in the Lepidoptera

Classic morphological studies of the oldest, so‐called nonditrysian lineages of Lepidoptera yielded a well‐resolved phylogeny, supported by the stepwise origin of the traits characterizing the clade Ditrysia, which contains over 98% of extant lepidopterans. Subsequent polymerase chain reaction (PCR)‐based molecular studies have robustly supported many aspects of the morphological hypothesis and strongly contradicted others, while leaving some relationships unsettled. Here we bring the greatly expanded gene sampling of RNA‐Seq to bear on nonditrysian phylogeny, especially those aspects that were not conclusively resolved by the combination of morphology and previous PCR‐based multi‐gene studies. We analysed up to 2212 genes in each of 28 species representing all 12 superfamilies and 15 of 21 families of nonditrysians, plus trichopteran outgroups and representative Ditrysia. Our maximum likelihood phylogeny estimates used both nonsynonymous changes only (degen1 coding) and all nucleotides (nt123) partitioned by codon position, recovering a novel hypothesis for early glossatan relationships that is the most strongly supported to date. We find strong support for Micropterigidae alone as the sister group to all other Lepidoptera, in agreement with morphology and early molecular evidence, but in contrast to recent PCR‐based studies. Also very strongly supported are the previously recognized clades Angiospermivora, Heteroneura, Eulepidoptera and Euheteroneura. Finally, we find strong support for paraphyly of the southern hemisphere family Palaephatidae, with the South American genus Palaephatus Butler forming the previously undetermined sister group to Ditrysia. The remaining palaephatids, Australian and South American, form the sister group to Tischeriidae.