Maria Heikkilä

Cretaceous origin and repeated tertiary diversification of the redefined butterflies

Although the taxonomy of the ca 18 000 species of butterflies and skippers is well known, the family-level relationships are still debated. Here, we present, to our knowledge, the most comprehensive phylogenetic analysis of the superfamilies Papilionoidea, Hesperioidea and Hedyloidea to date based on morphological and molecular data. We reconstructed their phylogenetic relationships using parsimony and Bayesian approaches. We estimated times and rates of diversification along lineages in order to reconstruct their evolutionary history. Our results suggest that the butterflies, as traditionally understood, are paraphyletic, with Papilionidae being the sister-group to Hesperioidea, Hedyloidea and all other butterflies. Hence, the families in the current three superfamilies should be placed in a single superfamily Papilionoidea. In addition, we find that Hedylidae is sister to Hesperiidae, and this novel relationship is supported by two morphological characters. The families diverged in the Early Cretaceous but diversified after the Cretaceous–Palaeogene event. The diversification of butterflies is characterized by a slow speciation rate in the lineage leading to Baronia brevicornis, a period of stasis by the skippers after divergence and a burst of diversification in the lineages leading to Nymphalidae, Riodinidae and Lycaenidae.

Morphology reinforces proposed molecular phylogenetic affinities: a revised classification for Gelechioidea (Lepidoptera)

Gelechioidea are one of the most species rich and least studied superfamilies of Lepidoptera. We examine the interrelationships within the superfamily using the densest taxon sampling to date, combined with the most extensive ever morphological and molecular character data. We perform partitioned and combined analyses using maximum likelihood, Bayesian and parsimony approaches. The combined dataset consists of 155 exemplar species of Gelechioidea, representing nearly all subfamilies recognized in recent classifications. Parsimony analyses are performed with a dataset including 28 additional terminal taxa with only morphological data available. We use eight genes with a total of 6127 bp, and morphological data with 253 characters derived from larval, pupal, and adult morphology. The analyses of combined data yield more resolved trees and significantly better‐supported groupings than either dataset when analysed alone. The recurrent monophyletic groupings in all our model‐based analyses support a revision of the family classification. Deeper relationships vary between analyses and data partitions, leaving them ambiguous. The place of the root remains a challenge for future research. We propose a revised classification and suggest the division of Gelechioidea into 16 families. We redefine Depressariidae Meyrick, 1883 for a monophylum that includes Acriinae, Aeolanthinae, Cryptolechiinae, Depressariinae, Ethmiinae, Hypercalliinae, Hypertrophinae, Peleopodinae, Oditinae, Stenomatinae, Carcina, and a diversity of predominantly New World taxa previously excluded from Lypusidae (Amphisbatidae s. authors) but left without family position. A monophyletic Oecophoridae s. s., including Deuterogoniinae and Pleurotinae, is obtained for the first time with significant support. Elachistidae s. l. is found to be polyphyletic, and Elachistidae is restricted to comprise Agonoxeninae, Elachistinae, and Parametriotinae. Batrachedridae are polyphyletic, with several genera pending further study. Apart from the core Batrachedra, the taxa previously included in this family are grouped in an expanded Pterolonchidae, together with Coelopoetinae and Syringopainae. Lypusidae s. s. and Chimabachidae form a monophylum; Chimabachinae is united with Lypusidae as a subfamily, stat. n. Our results contradict the subfamily classifications of several families, notably Lecithoceridae and Autostichidae, but due to insufficient sampling of taxa we refrain from comprehensive taxonomic conclusions on the subfamily level, and encourage focused studies to resolve these groups.

Elusive ditrysian phylogeny: an account of combining systematized morphology with molecular data (Lepidoptera)

BackgroundDitrysia comprise close to 99 % of all butterflies and moths. The evolutionary relationships among the ditrysian superfamilies have received considerable attention in phylogenetic studies based on DNA and transcriptomic data, but the deepest divergences remain for large parts unresolved or contradictory. To obtain complementary insight into the evolutionary history of the clade, and to test previous hypotheses on the subdivision of Ditrysia based on morphology, we examine the morphology of larvae, pupae and adult males and females of 318 taxa representing nearly all ditrysian superfamilies and families. We present the most comprehensive morphological dataset on Ditrysia to date, consisting of over 500 morphological characters. The data are analyzed alone and combined with sequence data (one mitochondrial and seven nuclear protein-coding gene regions, sequenced from 422 taxa). The full dataset consists of 473 exemplar species. Analyses are performed using maximum likelihood methods, and parsimony methods for the morphological dataset. We explore whether combining morphological data and DNA-data can stabilize taxa that are unstable in phylogenetic studies based on genetic data only.ResultsMorphological characters are found phylogenetically informative in resolving apical nodes (superfamilies and families), but characters serving as evidence of relatedness of larger assemblages are few. Results include the recovery of a monophyletic Tineoidea, Sesioidea and Cossoidea, and a stable position for some unstable taxa (e.g. Epipyropidae, Cyclotornidae, Urodoidea + Schreckensteinioidea). Several such taxa, however, remain unstable even though morphological characters indicate a position in the tree (e.g. Immidae). Evidence supporting affinities between clades are suggested, e.g. a novel larval synapomorphy for Tineidae. We also propose the synonymy of Tineodidae with Alucitidae, syn. nov.ConclusionsThe large morphological dataset provides information on the diversity and distribution of morphological traits in Ditrysia, and can be used in future research on the evolution of these traits, in identification keys and in identification of fossil Lepidoptera. The “backbone” of the phylogeny for Ditrysia remains largely unresolved. As previously proposed as an explanation for the scarcity of molecular signal in resolving the deeper nodes, this may be due to the rapid radiation of Ditrysia in the Cretaceous.

Phylogeny and feeding trait evolution of the mega-diverse Gelechioidea (Lepidoptera: Obtectomera): new insight from 19 nuclear genes

The Gelechioidea (>18 000 species), one of the largest superfamilies of Lepidoptera, are a major element of terrestrial ecosystems and include important pests and biological model species. Despite much recent progress, our understanding of the classification, phylogeny and evolution of Gelechioidea remains limited. Building on recent molecular studies of this superfamily and a recently revised family/subfamily classification, we provide an independent estimate of among‐family relationships, with little overlap in gene sample. We analysed up to five nuclear genes, totalling 6633 bp, for each of 77 gelechioids, plus up to 14 additional genes, for a total of 14 826 bp, in 45 of those taxa and all 19 outgroup taxa. Our maximum‐likelihood (ML) analyses, like those of previous authors, strongly support monophyly for most multiply‐sampled families and subfamilies, but very weakly support most relationships above the family level. Our tree looks superficially divergent from that of the most recent molecular study of gelechioids, but when the previous tree is re‐rooted to accord maximally with ours, the two phylogenies agree entirely on the deepest‐level divergences in Gelechioidea, and strongly though incompletely on among‐family relationships within the major groups. This concordance between independent studies is evidence that the groupings (or at least the unrooted branching order) are probably accurate, despite the low bootstrap values. After re‐rooting, both trees divide the families into three monophyletic groups: a ‘Gelechiid Assemblage,’ consisting of Gelechiidae and Cosmopterigidae; a ‘Scythridid Assemblage,’ consisting of Stathmopodidae, Scythrididae, Blastobasidae, Elachistidae, Momphidae, Coleophoridae and Batrachedridae; and a ‘Depressariid Assemblage,’ consisting of Autostichidae, Xyloryctidae, Lecithoceridae, Oecophoridae, Depressariidae and Lypusidae. Within the largest family, Gelechiidae, our results strongly support the pairing of Anomologinae with Gelechiinae, in accordance with a recent study of this family. Relationships among the other subfamilies, however, conflict moderately to strongly between studies, leaving the intrafamily phylogeny unsettled. Within the ‘Scythridid Assemblage,’ both trees support an ‘SSB clade’ consisting of Blastobasidae + (Scythrididae + Stathmopodidae), strongly resolved only in our results. Coleophoridae + Batrachedridae is supported, albeit weakly, in both trees, and only Momphidae differ in position between studies. Within the ‘Depressariid Assemblage,’ both trees support an ‘AXLO’ clade consisting of Autostichidae, Xyloryctidae, Lecithoceridae and Oecophoridae. The monophyly of this clade and relationships therein are supported weakly in previous results but strongly in ours. The recently re‐defined family Depressariidae is paraphyletic in our tree, but the evidence against depressariid monophyly is very weak. There is moderate support for a core group of Depressariidae consisting, among the seven subfamilies we sampled, of Depressariinae, Aeolanthinae and Hypertrophinae. We show that gelechioids have a higher total number and percentage of species that are saprophagous as larvae than any other apoditrysian superfamily, that saprophagy is concentrated primarily in the ‘AXLO clade,’ and that the ancestral gelechioid condition was probably feeding on live plants. Among the living‐plant feeders, concealed external feeding was probably the ancestral state. The multiple origins of internal feeding of various kinds, including leaf mining (otherwise almost unknown in Apoditrysia), are restricted mostly to the Scythridid and Gelechiid Assemblages. The traits that predispose or permit lineages to adopt these unusual life histories are worthy of study.

Reassessment of the enigmatic Lepidopteran family Lypusidae (Lepidoptera: Tineoidea; Gelechioidea).

Abstract. The genus Lypusa Zeller, 1852 has been assigned to Tineoidea (Lepidoptera) with varying family positions. The systematic affinities of this genus, currently placed in its own family Lypusidae, were studied using extensive data derived from larval, pupal and adult morphology and certain behavioural traits. In total, 193 characters were considered. On the basis of the results of a parsimony analysis, Lypusa is transferred to the superfamily Gelechioidea, in a monophyletic, exclusively Palaearctic assemblage with the genera Amphisbatis and Pseudatemelia. Several phylogenetically relevant characters support this position, including similarly constructed larval cases, densely porose larval head, and a modification of the pupal abdominal segment 8. The composition of the family Amphisbatidae is discussed and it is proposed that it be delimited as comprising solely the Palaearctic genera Pseudatemelia, Amphisbatis and Lypusa. A detailed diagnosis of the genus Lypusa is given.

The Trials and Tribulations of Priors and Posteriors in Bayesian Timing of Divergence Analyses: the Age of Butterflies Revisited

The need for robust estimates of times of divergence is essential for downstream analyses, yet assessing this robustness is still rare. We generated a time-calibrated genus-level phylogeny of butterflies (Papilionoidea), including 994 taxa, up to 10 gene fragments and an unprecedented set of 12 fossils and 10 host-plant node calibration points. We compared marginal priors and posterior distributions to assess the relative importance of the former on the latter. This approach revealed a strong influence of the set of priors on the root age but for most calibrated nodes posterior distributions shifted from the marginal prior, indicating significant information in the molecular dataset. We also tested the effects of changing assumptions for fossil calibration priors and the tree prior. Using a very conservative approach we estimated an origin of butterflies at 107.6 Ma, approximately equivalent to the Early Cretaceous–Late Cretaceous boundary, with a credibility interval ranging from 89.5 Ma (mid Late Cretaceous) to 129.5 Ma (mid Early Cretaceous). This estimate was robust to alternative analyses changing core assumptions. With 994 genera, this tree provides a comprehensive source of secondary calibrations for studies on butterflies.

Re-examining the rare and the lost: a review of fossil Tortricidae (Lepidoptera)

We re-evaluate eleven fossils that have previously been assigned to the family Tortricidae, describe one additional fossil, and assess whether observable morphological features warrant confident assignment of these specimens to this family. We provide an overview of the age and origin of the fossils and comment on their contribution towards understanding the phylogeny of the Lepidoptera. Our results show that only one specimen, Antiquatortia histuroides Brown Baixeras gen. and sp. nov., shows a character considered synapomorphic for the family. Six other fossils (Electresia zalesskii Kusnezov, 1941; Tortricidrosis inclusa Skalski, 1973; Tortricites skalskii Kozlov, 1988; Tortricibaltia diakonoffi Skalski, 1992; Polyvena horatis Poinar and Brown, 1993 and a trace fossil purported to be larval feeding damage of Retinia resinella (Linnaeus, 1758)) exhibit a combination of homoplastic characters typical of tortricid moths or characteristic feeding damage. An unnamed species doubtfully assigned to Olethreutinae by Skalski (1992), Spatalistiforma submerga Skalski, 1992, Tortricites florissantanus (Cockerell, 1907), Tortricites destructus (Cockerell, 1916) and Tortricites sadilenkoi Kozlov, 1988 do not show enough character evidence to be convincingly placed in Tortricidae. Therefore, we transfer the three latter species from the collective group Tortricites Kozlov, 1988, defined as an assemblage of fossil leafrollers that cannot be placed with certainty in known genera, to Paleolepidopterites Kozlov, new collective group, defined as a group of fossil lepidopterans whose assignment to a certain family is currently impossible.

Development of a nutrition knowledge questionnaire for young endurance athletes and their coaches

Both athletes and coaches should have adequate nutrition knowledge to understand the importance of diet on athletic performance, recovery, and health. Nutrition knowledge can be assessed reliably only by validated knowledge questionnaires. The aim of this study was to develop a reliable and valid questionnaire for assessing the nutrition knowledge of young endurance athletes and their coaches. The questionnaire was developed with an expert panel and pilot tested by athletes, coaches, and students. Content, face, and construct validities both as test‐retest reliability and internal consistency reliability were ensured when the current questionnaire was developed. Athletes (n = 16) and coaches (n = 13) pilot tested the 127‐item questionnaire. After item analysis and proposals from the expert panel, 41 items were removed. Internal consistency of the 86‐item questionnaire in the pilot study was 0.87, measured using Cronbachs α. Construct validity was evaluated by the difference in knowledge between nutrition (n = 20) and humanities students (n = 22). Nutrition students had significantly higher knowledge scores (P < .001). Test‐retest reliability for all knowledge sections between those groups was 0.85 measured using Pearsons r. Final adjustments to the questionnaire were made on the grounds of feedback from the respondents and proposals from the experts (n = 6). These adjustments resulted in minor changes in the construct of the items, the layout of the questionnaire, and the removal of 7 items. The final questionnaire had 79 items. The questionnaire can be used to measure the overall nutrition knowledge of endurance athletes and their coaches and to find potential gaps in nutrition knowledge.

Three New Species of Rectiostoma Becker, 1982 (Lepidoptera: Gelechioidea: Depressariidae) from Area De Conservación Guanacaste, Northwestern Costa Rica

Abstract. We describe three new species of Rectiostoma Becker, 1982 from northwestern Costa Rica: R. annemayae Heikkilä and Metz, n. sp., R. eowilsoni Heikkilä and Metz, n. sp., and R. philipmayi Heikkilä and Metz, n. sp. We used a data set of DNA barcodes (cytochrome oxidase I) accumulated for Lepidoptera collected at Area de Conservación Guanacaste (ACG) to initially delimit the species, then we confirmed identities with morphology of the male and female genitalia. We provide a neighbor-joining tree with supporting statistics, illustrations of adult specimens and their genitalia, and the first host plant records for Neotropical Rectiostoma species. We also include photographs of the larvae of R. annemayae and R. eowilsoni.

Figure 11 In Reassessment Of Known Fossil Pyraloidea (Lepidoptera) With Descriptions Of The Oldest Fossil Pyraloid And A Crambid Larva In Baltic Amber

The identifications of known fossils currently placed in the lepidopteran superfamily Pyraloidea are critically re-examined. Of the eleven fossils examined, only three are confirmed to show morphological characters supporting placement in the superfamily. These fossils include a crambid larva in Baltic Amber, Baltianania yantarnia, Solis gen. n. et sp. n. and the oldest known fossil pyraloid, Eopyralis morsae Simonsen, gen. n. et sp. n. The third fossil, Glendotricha olgae Kusnezov, 1941, displays apomorphic characters for Pyraloidea, but is shown to be an inclusion in copal, not Baltic amber as had been reported. Seven fossil specimens lack reliable characters and cannot be assigned to Pyraloidea with certainty: Pyralites obscurus Heer, 1856; Pyralites preecei Jarzembowski, 1980; Petisca dryellina Martins-Neto, 1998; three fossil larvae tentatively identified as Pyralidae by Zeuner (1931); and Gallerites keleri Kernbach, 1967. A possible fossil pyraloid in Mizunami amber could not be located in museum collections and available literature does not provide details to assess the validity of the identification. We discuss the contribution of the reliably identified fossils towards better understanding the evolutionary history of Pyraloidea.