Species-Level Para- and Polyphyly in DNA Barcode Gene Trees: Strong Operational Bias in European Lepidoptera.

The proliferation of DNA data is revolutionizing all fields of systematic research. DNA barcode sequences, now available for millions of specimens and several hundred thousand species, are increasingly used in algorithmic species delimitations. This is complicated by occasional incongruences between species and gene genealogies, as indicated by situations where conspecific individuals do not form a monophyletic cluster in a gene tree. In two previous reviews, non-monophyly has been reported as being common in mitochondrial DNA gene trees. We developed a novel web service "Monophylizer" to detect non-monophyly in phylogenetic trees and used it to ascertain the incidence of species non-monophyly in COI (a.k.a. cox1) barcode sequence data from 4977 species and 41,583 specimens of European Lepidoptera, the largest data set of DNA barcodes analyzed from this regard. Particular attention was paid to accurate species identification to ensure data integrity. We investigated the effects of tree-building method, sampling effort, and other methodological issues, all of which can influence estimates of non-monophyly. We found a 12% incidence of non-monophyly, a value significantly lower than that observed in previous studies. Neighbor joining (NJ) and maximum likelihood (ML) methods yielded almost equal numbers of non-monophyletic species, but 24.1% of these cases of non-monophyly were only found by one of these methods. Non-monophyletic species tend to show either low genetic distances to their nearest neighbors or exceptionally high levels of intraspecific variability. Cases of polyphyly in COI trees arising as a result of deep intraspecific divergence are negligible, as the detected cases reflected misidentifications or methodological errors. Taking into consideration variation in sampling effort, we estimate that the true incidence of non-monophyly is ∼23%, but with operational factors still being included. Within the operational factors, we separately assessed the frequency of taxonomic limitations (presence of overlooked cryptic and oversplit species) and identification uncertainties. We observed that operational factors are potentially present in more than half (58.6%) of the detected cases of non-monophyly. Furthermore, we observed that in about 20% of non-monophyletic species and entangled species, the lineages involved are either allopatric or parapatric-conditions where species delimitation is inherently subjective and particularly dependent on the species concept that has been adopted. These observations suggest that species-level non-monophyly in COI gene trees is less common than previously supposed, with many cases reflecting misidentifications, the subjectivity of species delimitation or other operational factors.

Revision of the genus Pollanisus Walker, 1854(Lepidoptera: Zygaenidae: Procridinae).

The genus Pollanisus is endemic for Australia. Its revision is mainly based on head studies and includes 21 known species and 7 new species, Pollanisus jumbun sp. n., Pollanisus yugambeh sp. n., Pollanisus horakae sp. n., Pollanisus worimi sp. n., Pollanisus kalliesi sp. n., Pollanisus jirrbal sp. n. and Pollanisus nocturna sp. n. Moreover, two species, Pollanisus eumetopus syn. n. and Pollanisus eungellae syn. n., are synonymized with Pollanisus acharon (Fabricius, 1775). The status of 8 unnamed species (Tarmann, 2004) is discussed. New data on the phenology and bionomics are provided. New host plants in the genus Hibbertia (Dilleniaceae) are reported.

A new species of Piarosoma Hampson, 1893 (Lepidoptera, Zygaenidae, Procridinae, Artonini) from Arunachal Pradesh, India.

A new species, Piarosoma arunachalensis sp. nov., is described and illustrated based on external characters and the genitalia of two male specimens from Tale Wildlife Sanctuary, Lower Subansiri District, Arunachal Pradesh, India. The new species is distinguished from other Piarosoma by a combination of: a) presence of large hyaline patches on the wings, b) the absence of any white or yellow markings on the patagia and the abdominal segments, and c) differences in male genitalia.

Drastic decline of extensive grassland species in Central Europe since 1950: Forester moths of the genus Jordanita (Lepidoptera, Zygaenidae) as a type example.

The decline of biodiversity in general and of insect diversity in particular has been recognized as a major environmental problem in recent years. In this study, we analyze the distribution and the decline of populations of forester moths of the genus Jordanita in Central Europe since 1950 as a type example of the loss of grassland biodiversity, and discuss potential drivers causing this decline. Based on the extensive work in museums and private collections, a literature review and own observations, and including data as far back as 1834, this genus helps to understand the deeper reasons of insect population and biodiversity decline, as the well investigated six Central European species cover a broad range of extensive grassland habitats (fens to low-production grassland and xerothermic steppes) from low altitudes to high alpine meadows. Therefore, they monitor processes relevant also to other, less investigated grassland species. Although there are differences in research intensity over time and in different natural areas, we show that in the whole of Central Europe, the populations of all six investigated Jordanita species broke down massively in the past decades, both in terms of number of populated habitats (about 400 recorded localities after the year 2000 compared with a total number of about 1600 at all times, cumulated for all six species) and in terms of number of individuals. On the other hand, some natural areas on a regional scale have more or less maintained their Jordanita populations, due to conservative land use and due to the early implementation of conservation and protection management plans. The reasons of decline are manifold and monitored in detail by the different species with their different habitat requirements. They comprise (1) loss of habitats due to land use changes (both intensification and abandonment), (2) loss of habitats due to urbanization and construction work, (3) loss of habitat networks to cope with small-scale extinction events, (4) more intensive growth of grass at the expense of other plants in otherwise undisturbed habitats due to fertilization through the air (increased nitrogen levels due to human activities) and (5) use of pesticides.

Phylogenetic position of the 'extinct' Fijian coconut moth, Levuana iridescens (Lepidoptera: Zygaenidae).

Levuana iridescens Bethune-Baker, 1906, a day-flying moth purported to be endemic to the Fijian Island of Viti Levu and a former pest of its coconut palm trees, was last observed in 1956 and has been officially declared extinct by IUCN since 1996. The controversial classical biological control method that resulted in the (presumed) demise of this moth has given this species an iconic status in biological control studies. We investigated the sister-group relationships and phylogenetic placement of this moth using NGS-obtained ancient DNA sequences from museum specimens of L. iridescens collected in the 1920s, combined with 31 morphological characters used in earlier studies and 2 new characters. Our findings show that Levuana is most closely related to the Australian genus Myrtartona. The significance of these findings is discussed.

"Ino Budensis var. Mollis" Grum-Grshimailo, 1893 (Lepidoptera: Zygaenidae) from Eastern Asia recognized as a valid species on the base of morphological and molecular analysis.

Based on morphological and molecular data, Ino budensis var. mollis Grum-Grshimailo, 1893, from China, so far treated as a synonym of Jordanita (Roccia) paupera (Christoph, 1887), is here recognized as a good species, Jordanita (Roccia) mollis (Grum-Grshimailo, 1893), stat. nov. This species is recorded as new for the fauna of Russia and Korea. An identification key for this species is provided.