A new North American species of Etainia (Lepidoptera, Nepticulidae), feeding on Arbutus and Arctostaphylos species (Ericaceae).

Etainiathoraceleuca van Nieukerken, Epstein & Davis, sp. nov. is the second native American species of Etainia Beirne, 1945, and the second known Etainia species feeding on Ericaceae. The species is known from light-collected adults in the USA (California, Arizona) and Canada (Ontario). These were linked via DNA barcodes to larvae that make short leafmines on Arbutus and Arctostaphylos species, then continue feeding in stems and branches, causing damage in nurseries and planted trees in Sonoma and Marin Counties, California. The holotype was accidentally reared from Arbutusarizonica, without observing the damage. Life history and damage are described in detail. Damage in Arctostaphylosuva-ursi found in Washington State probably belongs to E.thoraceleuca, which is a sister species to the European E.albibimaculella (Larsen, 1927).

Poleward increase in feeding efficiency of leafminer Stigmella lapponica (Lepidoptera: Nepticulidae) in a latitudinal gradient crossing a boreal forest zone.

Damage to plant communities imposed by insect herbivores generally decreases from low to high latitudes. This decrease is routinely attributed to declines in herbivore abundance and/or diversity, whereas latitudinal changes in per capita food consumption remain virtually unknown. Here, we tested the hypothesis that the lifetime food consumption by a herbivore individual decreases from low to high latitudes due to a temperature-driven decrease in metabolic expenses. From 2016 to 2019, we explored latitudinal changes in multiple characteristics of linear (gallery) mines made by larvae of the pygmy moth, Stigmella lapponica, in leaves of downy birch, Betula pubescens. The mined leaves were larger than intact leaves at the southern end of our latitudinal gradient (at 60°N) but smaller than intact leaves at its northern end (at 69°N), suggesting that female oviposition preference changes with latitude. No latitudinal changes were observed in larval size, mine length or area, and in per capita food consumption, but the larval feeding efficiency (quantified as the ratio between larval size and mine size) increased with latitude. Consequently, S. lapponica larvae consumed less foliar biomass at higher latitudes than at lower latitudes to reach the same size. Based on space-for-time substitution, we suggest that climate warming will increase metabolic expenses of insect herbivores with uncertain consequences for plant-herbivore interactions.

Phylogeny of gracillariid leaf-mining moths: evolution of larval behaviour inferred from phylogenomic and Sanger data.

Gracillariidae is the most taxonomically diverse cosmopolitan leaf-mining moth family, consisting of nearly 2000 named species in 105 described genera, classified into eight extant subfamilies. The majority of gracillariid species are internal plant feeders as larvae, creating mines and galls in plant tissue. Despite their diversity and ecological adaptations, their phylogenetic relationships, especially among subfamilies, remain uncertain. Genomic data (83 taxa, 589 loci) were integrated with Sanger data (130 taxa, 22 loci), to reconstruct a phylogeny of Gracillariidae. Based on analyses of both datasets combined and analyzed separately, monophyly of Gracillariidae and all its subfamilies, monophyly of the clade "LAMPO" (subfamilies: Lithocolletinae, Acrocercopinae, Marmarinae, Phyllocnistinae, and Oecophyllembiinae) and relationships of its subclade "AMO" (subfamilies: Acrocercopinae, Marmarinae, and Oecophyllembiinae) were strongly supported. A sister-group relationship of Ornixolinae to the remainder of the family, and a monophyletic leaf roller lineage (Callicercops Vári + Parornichinae) + Gracillariinae, as sister to the "LAMPO" clade were supported by the most likely tree. Dating analyses indicate a mid-Cretaceous (105.3 Ma) origin of the family, followed by a rapid diversification into the nine subfamilies predating the Cretaceous-Palaeogene extinction. We hypothesize that advanced larval behaviours, such as making keeled or tentiform blotch mines, rolling leaves and galling, allowed gracillariids to better avoid larval parasitoids allowing them to further diversify. Finally, we stabilize the classification by formally re-establishing the subfamily ranks of Marmarinae stat.rev., Oecophyllembiinae stat.rev. and Parornichinae stat.rev., and erect a new subfamily, Callicercopinae Li, Ohshima and Kawahara to accommodate the enigmatic genus Callicercops.

Identity of some recently described Lepidoptera from France-re-assessed with DNA barcodes and morphology.

Seventy-three species of Lepidoptera described from France since 2000, particularly by Jacques Nel and Thierry Varenne, are re-assessed from largely unpublished molecular data. We tried to obtain DNA barcode sequences from 62 holotypes, supplemented by paratypes of eight species and on one case by non-type material, whereas one previously synonymized species was not sequenced. Altogether we obtained 78 DNA barcode sequences for 65 nominal taxa while sequencing failed for six holotypes. An integrative analysis from molecular data and morphology supports the validity of the majority of species but also resulted in the re-assessment of several taxa. The following 13 new synonymies are established: Stigmella cyrneorolandi Nel Varenne, 2013 syn. nov. of Stigmella rolandi van Nieukerken, 1990; Stigmella thibaulti Varenne Nel, 2019 syn. nov. of Stigmella nivenburgensis (Preissecker, 1942) (Nepticulidae); Nemapogon peslieri Varenne Nel, 2017 syn. nov. of Nemapogon inexpectata Varenne Nel, 2017 (Tineidae); Phyllonorycter acericorsica Varenne Nel, 2015 syn. nov. of Phyllonorycter ochreojunctella (Klimesch, 1942) (Gracillariidae); Ancylis paraobtusana Varenne, Nel, Peslier, 2020 syn. nov. of Ancylis comptana (Frölich, 1828) (Tortricidae); Celypha paludicolella Varenne Nel, 2017 syn. nov. of Celypha doubledayana (Barrett, 1872) (Tortricidae); Cydia oxytropidana Nel Varenne, 2016 syn. nov. of Cydia oxytropidis (Martini, 1912) (Tortricidae); Sorhagenia orocorsa Varenne Nel, 2016 syn. nov. of Sorhagenia janiszewskae Riedl, 1962 (Cosmopterigidae); Chionodes cerdanica Peslier, Nel Varenne, 2020 syn. nov. of Chionodes distinctella (Zeller, 1839) (Gelechiidae); Elachista bidentata Varenne Nel, 2019 syn. nov. of Elachista orstadii Palm, 1943; Elachista karsticola Varenne Nel, 2018 syn. nov. of Elachista maculosella Chrétien, 1896 (Elachistidae); Scythris chablaisensis Delmas, 2018 syn. nov. of Scythris laminella ([Denis Schiffermüller], 1775) (Scythrididae); Epermenia pumila (Buvat Nel, 2000) syn. nov. of Epermenia profugella (Stainton, 1856) (Epermeniidae). Finally, the status of some taxa still remains unclear due to the lack of DNA barcodes of closely related species and the absence of convincing diagnostic characters in morphology.

Splitting the leafmining shield-bearer moth genus Antispila Hübner (Lepidoptera, Heliozelidae): North American species with reduced venation placed in Aspilanta new genus, with a review of heliozelid morphology.

The new genus Aspilanta gen. n. is described to harbour Nearctic heliozelid moths with reduced venation, previously placed in Antispila Hübner, 1825, with type species Antispila oinophylla van Nieukerken & Wagner, 2012. The erection of this genus has become possible now that monophyly has been supported by a recent phylotranscriptomics analysis. Six species are combined in this genus: Aspilanta oinophylla (van Nieukerken & Wagner, 2012), comb. n., A. hydrangaeella (Chambers, 1874), comb. n., A. ampelopsifoliella (Chambers, 1874), comb. n., A. voraginella (Braun, 1927), comb. n., A. argentifera (Braun, 1927), comb. n., A. viticordifoliella (Clemens, 1860), comb. n. and two candidate species are recognised. DNA barcode COI sequences of Malaise trapped specimens suggest a rich fauna of Aspilanta in Central America. All are leafminers, with Vitaceae as main host family, and single species feeding respectively on Hydrangeaceae and Myricaceae. The species are briefly diagnosed, and data on biology, DNA barcodes and distribution are provided. To place the genus in context, a review of heliozelid morphology and phylogeny is presented and a key to Nearctic genera is given. The genus is confined to North and Central America, possibly also occurring in South America. Aspilanta oinophylla is also an invasive species on grapevine in Italy. The genus is sister to Coptodisca Walsingham, 1895. Another species is removed from Antispila: Heliozela eugeniella (Busck, 1900), comb. n., feeding on Eugenia (Myrtaceae), from Florida.

Sex Pheromones of Two Leafminer Species, Antispila oinophylla and Holocacista rivillei (Lepidoptera: Heliozelidae) Infesting Grapevine in Italy.

Two heliozelid species, Antispila oinophylla van Nieukerken & Wagner and Holocacista rivillei (Stainton) severely infest Italian grapevines. The volatile pheromones from calling females were collected by solid phase micro extraction (SPME) and analyzed by gas chromatography with electroantennographic detection (GC-EAD). Two compounds from A. oinophylla females eliciting electrophysiological activity from the conspecific male antenna were identified as (Z)-5-tetradecenal and (Z)-7-tetradecenal by coupled gas chromatography/mass spectrometry (GC/MS) analysis. SPME collections from H. rivillei produced no GC-EAD active compounds but analysis of fatty acyl moieties in the pheromone gland, demonstrated the presence of the putative pheromone biosynthetic precursors (Z)-5-dodecenoic acid and (Z)-7-tetradecenoic acid. Field trapping experiments in Italy confirmed that (Z)-5-tetradecenal and (Z)-7-tetradecenal are essential for the attraction of male A. oinophylla in a blend ratio of 15:100 respectively, whereas (Z)-5-dodecenal and (Z)-7-tetradecenal attract male H. rivillei in a blend ratio of 100:6.

Stigmellamultispicata Rociene. & Stonis, an Asian leafminer on Siberian elm, now widespread in eastern North America (Lepidoptera, Nepticulidae).

Stigmellamultispicata Rociene & Stonis, 2014, previously known from the single male holotype from Primorye, Russia, is reported as a new invasive species mining leaves of Siberian elm, Ulmuspumila L., in eastern North America. Both adults and leafmines have been reported from many sites as unidentified Nepticulidae since 2010. Crucial for the identification was a match of the DNA barcode of a single larva collected on Ulmuspumila in Beijing with adults from North America. The single larva constitutes a new record for China. Stigmellamultispicata is closely related to the European S.ulmivora (Fologne, 1860), feeding likewise on Ulmus, but differs in details of external morphology and genitalia, particularly in the female, where S.multispicata has a remarkable elongated narrow ovipositor, suitable for oviposition in underside hairy leaf vein axils, where all mines start. In North America S.multispicata is the only Ulmus-feeding nepticulid with green larvae. Currently the species is known from USA: Illinois, Indiana, Iowa, Maryland, Massachusetts, Minnesota, New York, Ohio, Tennessee, Wisconsin, and Canada: Ontario and Québec. In Sagaponack, on Long Island, New York, larvae have been reported to occur en masse on Siberian elms from at least two sites. The current distribution could be reconstructed thanks also to many online photographs from observation websites. The species is redescribed, with the first descriptions of female, larva, and leafmine, and compared with S.ulmivora, which is fully redescribed. The two native North American nepticulid Ulmus leafminers, S.apicialbella (Chambers, 1873) and Ectoedemiaulmella (Braun, 1912), are diagnosed and new provincial and state records are provided. A key to linear mines on Ulmus in North America is provided. We suspect that trade of live plants through nurseries played a role in the sudden spread of this invasive species.

A preliminary molecular phylogeny of shield-bearer moths (Lepidoptera: Adeloidea: Heliozelidae) highlights rich undescribed diversity.

Heliozelidae are a widespread, evolutionarily early diverging family of small, day-flying monotrysian moths, for which a comprehensive phylogeny is lacking. We generated the first molecular phylogeny of the family using DNA sequences of two mitochondrial genes (COI and COII) and two nuclear genes (H3 and 28S) from 130 Heliozelidae specimens, including eight of the twelve known genera: Antispila, Antispilina, Coptodisca, Heliozela, Holocacista, Hoplophanes, Pseliastis, and Tyriozela. Our results provide strong support for five major Heliozelidae clades: (i) a large widespread clade containing the leaf-mining genera Antispilina, Coptodisca and Holocacista and some species of Antispila, (ii) a clade containing most of the described Antispila, (iii) a clade containing the leaf-mining genus Heliozela and the monotypic genus Tyriozela, (iv) an Australian clade containing Pseliastis and (v) an Australian clade containing Hoplophanes. Each clade includes several new species and potentially new genera. Collectively, our data uncover a rich and undescribed diversity that appears to be especially prevalent in Australia. Our work highlights the need for a major taxonomic revision of the family and for generating a robust molecular phylogeny using multi-gene approaches in order to resolve the relationships among clades.

New taxa, including three new genera show uniqueness of Neotropical Nepticulidae (Lepidoptera).

After finding distinct clades in a molecular phylogeny for Nepticulidae that could not be placed in any known genera and discovering clear apomorphic characters that define these clades, as well as a number of Neotropical species that could be placed in known genera but were undescribed, three new genera and nine new species are here described from the Neotropics: Stigmella gallicola van Nieukerken & Nishida, sp. n. reared from galls on Hampea appendiculata (Malvaceae) in Costa Rica, representing the first example of a gall making Stigmella; Stigmella schinivora van Nieukerken, sp. n. reared from leafmines on Schinus terebinthifolia (Anacardiaceae) in Argentina, Misiones; Stigmella costaricensis van Nieukerken & Nishida, sp. n. and Stigmella intronia van Nieukerken & Nishida, sp. n. each from a single specimen collected the same night in Costa Rica, Parque Nacional Chirripó; Stigmella molinensis van Nieukerken & Snyers, sp. n. reared from leafmines on Salix humboldtiana, Peru, Lima, the first Neotropical species of the Stigmella salicis group sensu stricto; Ozadelpha van Nieukerken, gen. n. with type species Ozadelpha conostegiae van Nieukerken & Nishida, sp. n., reared from leafmines on Conostegia oerstediana (Melastomataceae) from Costa Rica; Neotrifurcula van Nieukerken, gen. n. with type species Neotrifurcula gielisorum van Nieukerken, sp. n. from Chile; Hesperolyra van Nieukerken, gen. n.. with type species Fomoria diskusi Puplesis & Robinson, 2000; Hesperolyra saopaulensis van Nieukerken, sp. n., reared from an unidentified Myrtaceae, Sao Paulo, Brasil; and Acalyptris janzeni van Nieukerken & Nishida, sp. n. from Costa Rica, Guanacaste. Five new combinations are made: Ozadelpha ovata (Puplesis & Robinson, 2000), comb. n. and Ozadelpha guajavae (Puplesis & Diskus, 2002), comb. n., Hesperolyra diskusi (Puplesis & Robinson, 2000), comb. n., Hesperolyra molybditis (Zeller, 1877), comb. n. and Hesperolyra repanda (Puplesis & Diskus, 2002), comb. n. Three specimens are briefly described, but left unnamed: Ozadelpha specimen EvN4680, Neotrifurcula specimen EvN4504 and Neotrifurcula specimen RH2.

Revised classification and catalogue of global Nepticulidae and Opostegidae (Lepidoptera, Nepticuloidea).

A catalogue of all named Nepticulidae and Opostegidae is presented, including fossil species. The catalogue is simultaneously published online in the scratchpad http://nepticuloidea.info/ and in Catalogue of Life (http://www.catalogueoflife.org/col/details/database/id/172). We provide a historical overview of taxonomic research on Nepticuloidea and a brief 'state of the art'. A DNA barcode dataset with 3205 barcodes is made public at the same time, providing DNA barcodes of ca. 779 species, of which 2563 are identified as belonging to 444 validly published species. We recognise 862 extant and 18 fossil species of Nepticulidae in 22 extant genera and the fossil form genus Stigmellites. We count 192 valid Opostegidae species in 7 genera, without fossils. We also list seven dubious Nepticulidae names that cannot be placed due to absent type material and poor descriptions, 18 unavailable names in Nepticulidae that cannot be placed and we also list the 33 names (including four fossils) that once were placed as Nepticulidae or Opostegidae but are now excluded. All synonyms and previous combinations are listed. The generic classification follows the Molecular phylogeny that is published almost simultaneously. Subfamilies and tribes are not recognised, Trifurculinae Scoble, 1983 is synonymised with Nepticulidae Stainton, 1854 and Opostegoidinae Kozlov, 1987 is synonymised with Opostegidae Meyrick, 1893. The status of Casanovula Hoare, 2013, Etainia Beirne, 1945, Fomoria Beirne, 1945, Glaucolepis Braun, 1917, Menurella Hoare, 2013, Muhabbetana Koçak & Kemal, 2007 and Zimmermannia Hering, 1940 is changed from subgenus to full genus, whereas two genera are considered synonyms again: Manoneura Davis, 1979, a synonym of Enteucha Meyrick, 1915 and Levarchama Beirne, 1945, a synonym of Trifurcula Zeller, 1848. We propose 87 new combinations in Nepticulidae and 10 in Opostegidae, largely due to the new classification, and re-examination of some species. We propose the following 37 new synonymies for species (35 in Nepticulidae, 2 in Opostegidae): Stigmella acerifoliella Dovnar-Zapolski, 1969 (unavailable, = Stigmella acerna Puplesis, 1988), Stigmella nakamurai Kemperman & Wilkinson, 1985 (= Stigmella palionisi Puplesis, 1984), Nepticula amseli Skala, 1941 (unavailable = Stigmella birgittae Gustafsson, 1985), Stigmella cathepostis Kemperman & Wilkinson, 1985 (= Stigmella microtheriella (Stainton, 1854)), Stigmella populnea Kemperman & Wilkinson, 1985 (= Stigmella nivenburgensis (Preissecker, 1942)), Nepticula obscurella Braun, 1912 (revised synonymy, = Stigmella myricafoliella (Busck, 1900)), Nepticula mandingella Gustafsson, 1972 (= Stigmella wollofella (Gustafsson, 1972)), Stigmella rosaefoliella pectocatena Wilkinson & Scoble, 1979 (= Stigmella centifoliella (Zeller, 1848)), Micropteryx pomivorella Packard, 1870 (= Stigmella oxyacanthella (Stainton, 1854)), Stigmella crataegivora Puplesis, 1985 (= Stigmella micromelis Puplesis, 1985), Stigmella scinanella Wilkinson & Scoble, 1979 (= Stigmella purpuratella (Braun, 1917)), Stigmella palmatae Puplesis, 1984 (= Stigmella filipendulae (Wocke, 1871)), Stigmella sesplicata Kemperman & Wilkinson, 1985 (= Stigmella lediella (Schleich, 1867)), Stigmella rhododendrifolia Dovnar-Zapolski & Tomilova, 1978 (unavailable, = Stigmella lediella (Schleich, 1867)), Stigmella oa Kemperman & Wilkinson, 1985 (= Stigmella spiculifera Kemperman & Wilkinson, 1985), Stigmella gracilipae Hirano, 2014 (= Stigmella monticulella Puplesis, 1984), Nepticula chaoniella Herrich-Schäffer, 1863 (= Stigmella samiatella (Zeller, 1839)), Bohemannia piotra Puplesis, 1984 (= Bohemannia pulverosella (Stainton, 1849)), Bohemannia nipponicella Hirano, 2010 (= Bohemannia manschurella Puplesis, 1984), Sinopticula sinica Yang, 1989 (= Glaucolepis oishiella (Matsumura, 1931)), Trifurcula collinella Nel, 2012 (= Glaucolepis magna (A. Lastuvka & Z. Lastuvka, 1997)), Obrussa tigrinella Puplesis, 1985 (= Etainia trifasciata (Matsumura, 1931)), Microcalyptris vittatus Puplesis, 1984 and Microcalyptris arenosus Falkovitsh, 1986 (both = Acalyptris falkovitshi (Puplesis, 1984)), Ectoedemia castaneae Busck, 1913, Ectoedemia heinrichi Busck, 1914 and Ectoedemia helenella Wilkinson, 1981 (all three = Zimmermannia bosquella (Chambers, 1878)), Ectoedemia chloranthis Meyrick, 1928 and Ectoedemia acanthella Wilkinson & Newton, 1981 (both = Zimmermannia grandisella (Chambers, 1880)), Ectoedemia coruscella Wilkinson, 1981 (= Zimmermannia mesoloba (Davis, 1978)), Ectoedemia piperella Wilkinson & Newton, 1981 and Ectoedemia reneella Wilkinson, 1981 (both = Zimmermannia obrutella (Zeller, 1873)), Ectoedemia similigena Puplesis, 1994 (= Ectoedemia turbidella (Zeller, 1848)), Ectoedemia andrella Wilkinson, 1981 (= Ectoedemia ulmella (Braun, 1912)), Nepticula canadensis Braun, 1917 (= Ectoedemia minimella (Zetterstedt, 1839)), Opostega rezniki Kozlov, 1985 (= Opostega cretatella Chrétien, 1915), Pseudopostega cyrneochalcopepla Nel & Varenne, 2012 (= Pseudopostega chalcopepla (Walsingham, 1908)). Stigmella caryaefoliella (Clemens, 1861) and Zimmermannia bosquella (Chambers, 1878) are taken out of synonymy and re-instated as full species. Lectotypes are designated for Trifurcula obrutella Zeller, 1873 and Nepticula grandisella Chambers, 1880.

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.

Identification of the Female-Produced Sex Pheromone of the Leafminer Holocacista capensis Infesting Grapevine in South Africa.

We report the first identification of a sex pheromone in a heliozelid moth, Holocacista capensis van Nieukerken & Geertsema. This leafminer recently infested grapevine in South Africa. Compared to solvent extraction of pheromone glands, solid phase microextraction (SPME) proved to be highly effective for collection of the pheromone from calling females. The volatiles collected by SPME were analyzed by gas chromatography with electroantennographic detection (GC/EAD). Three compounds eliciting electrophysiological activity from the male antenna were identified as (Z)-5-tetradecenal, (Z)-7-tetradecenal, and (Z)-9-hexadecenal by coupled gas chromatography-mass spectrometry (GC/MS). GC/MS analysis of dimethyldisulphide (DMDS) derivatives of fatty acyl moieties in pheromone gland extracts confirmed the presence of the corresponding putative pheromone precursors with double bonds in the same position and with Z geometry. Field trapping experiments in a South African vineyard confirmed that both (Z)-5-tetradecenal and (Z)-7-tetradecenal are essential for the attraction of male H. capensis, whereas addition of (Z)-9-hexadecenal to the blend did not affect the attractiveness. The composition of the pheromone is discussed in relation to the phylogeny of this family of moths.

A new leafminer on grapevine and Rhoicissus (Vitaceae) in South Africa within an expanded generic concept of Holocacista (Insecta, Lepidoptera, Heliozelidae).

A grapevine leafminer found recently in table grape orchards and vineyards in the Paarl region (Western Cape, South Africa) is described as Holocacistacapensis sp. n. It has also been found on native Rhoicissusdigitata and bred on that species in the laboratory. It is closely related to Holocacistasalutans (Meyrick, 1921), comb. n. (from Antispila), described from Durban in KwaZulu-Natal, but widespread in southern Africa and a native leafminer of various Vitaceae: Rhoicissustomentosa, Rhoicissusdigitata, Rhoicissustridentata and Cissuscornifolia. Holocacistacapensis has been found on Vitisvinifera both in Gauteng and Western Cape, the earliest record being from 1950 in Pretoria. The initial host shift from native Vitaceae to Vitis must have occurred much earlier. The species is sometimes present in high densities, but hitherto no sizeable damage to the crops has been noted. The genus Holocacista Walsingham & Durrant, 1909, previously known from the single European grapevine leafminer Holocacistarivillei (Stainton, 1855), is expanded and redescribed and for the first time reported from Africa, East and South-East Asia and Australia. It comprises seven named species and at least 15 unnamed species. The following species are also recombined with Holocacista: transferred from Antispilina: South-African Holocacistavarii (Mey, 2011), comb. n., feeding on Pelargonium, transferred from Antispila: the Indian species Holocacistamicrarcha (Meyrick, 1926), comb. n. and Holocacistapariodelta (Meyrick, 1929), comb. n., both feeding on Lanneacoromandelica, and Holocacistaselastis (Meyrick, 1926), comb. n. on Psychotriadalzelii. We also remove the following from Antispila: Heliozelaanna (Fletcher, 1920), comb. n. and Heliozelaargyrozona (Meyrick, 1918), comb. n., whereas the following Indian Vitaceae feeding species are confirmed to belong in Antispila s. str.: Antispilaargostoma Meyrick, 1916 and Antispilaaristarcha Meyrick, 1916. Holocacistasalutans and Holocacistavarii are redescribed and diagnosed against Holocacistacapensis and other South African Heliozelidae. DNA barcodes are provided for 13 species of Holocacista.

A global phylogeny of leafmining Ectoedemia moths (Lepidoptera: Nepticulidae): exploring host plant family shifts and allopatry as drivers of speciation.

BACKGROUND: Host association patterns in Ectoedemia (Lepidoptera: Nepticulidae) are also encountered in other insect groups with intimate plant relationships, including a high degree of monophagy, a preference for ecologically dominant plant families (e.g. Fagaceae, Rosaceae, Salicaceae, and Betulaceae) and a tendency for related insect species to feed on related host plant species. The evolutionary processes underlying these patterns are only partly understood, we therefore assessed the role of allopatry and host plant family shifts in speciation within Ectoedemia. METHODOLOGY: Six nuclear and mitochondrial DNA markers with a total aligned length of 3692 base pairs were used to infer phylogenetic relationships among 92 species belonging to the subgenus Ectoedemia of the genus Ectoedemia, representing a thorough taxon sampling with a global coverage. The results support monophyletic species groups that are congruent with published findings based on morphology. We used the obtained phylogeny to explore host plant family association and geographical distribution to investigate if host shifts and allopatry have been instrumental in the speciation of these leafmining insects. SIGNIFICANCE: We found that, even though most species within species groups commonly feed on plants from one family, shifts to a distantly related host family have occasionally occurred throughout the phylogeny and such shifts are most commonly observed towards Betulaceae. The largest radiations have occurred within species groups that feed on Fagaceae, Rosaceae, and Salicaceae. Most species are restricted to one of the seven global biogeographic regions, but within species groups representatives are commonly found in different biogeographic regions. Although we find general patterns with regard to host use and biogeography, there are differences between clades that suggest that different drivers of speciation, and perhaps drivers that we did not examine, have shaped diversity patterns in different clades.

A Linnaeus NG (TM) interactive key to the Lithocolletinae of North-West Europe aimed at accelerating the accumulation of reliable biodiversity data (Lepidoptera, Gracillariidae).

We present an interactive key that is available online through any web browser without the need to install any additional software, making it an easily accessible tool for the larger public. The key can be found at http://identify.naturalis.nl/lithocolletinae. The key includes all 86 North-West European Lithocolletinae, a subfamily of smaller moths ("micro-moths") that is commonly not treated in field guides. The user can input data on several external morphological character systems in addition to distribution, host plant and even characteristics of the larval feeding traces to reach an identification. We expect that this will enable more people to contribute with reliable observation data on this group of moths and alleviate the workload of taxonomic specialists, allowing them to focus on other new keys or taxonomic work.

Genetic patterns in European geometrid moths revealed by the Barcode Index Number (BIN) system.

BACKGROUND: The geometrid moths of Europe are one of the best investigated insect groups in traditional taxonomy making them an ideal model group to test the accuracy of the Barcode Index Number (BIN) system of BOLD (Barcode of Life Datasystems), a method that supports automated, rapid species delineation and identification. METHODOLOGY/PRINCIPAL FINDINGS: This study provides a DNA barcode library for 219 of the 249 European geometrid moth species (88%) in five selected subfamilies. The data set includes COI sequences for 2130 specimens. Most species (93%) were found to possess diagnostic barcode sequences at the European level while only three species pairs (3%) were genetically indistinguishable in areas of sympatry. As a consequence, 97% of the European species we examined were unequivocally discriminated by barcodes within their natural areas of distribution. We found a 1:1 correspondence between BINs and traditionally recognized species for 67% of these species. Another 17% of the species (15 pairs, three triads) shared BINs, while specimens from the remaining species (18%) were divided among two or more BINs. Five of these species are mixtures, both sharing and splitting BINs. For 82% of the species with two or more BINs, the genetic splits involved allopatric populations, many of which have previously been hypothesized to represent distinct species or subspecies. CONCLUSIONS/SIGNIFICANCE: This study confirms the effectiveness of DNA barcoding as a tool for species identification and illustrates the potential of the BIN system to characterize formal genetic units independently of an existing classification. This suggests the system can be used to efficiently assess the biodiversity of large, poorly known assemblages of organisms. For the moths examined in this study, cases of discordance between traditionally recognized species and BINs arose from several causes including overlooked species, synonymy, and cases where DNA barcodes revealed regional variation of uncertain taxonomic significance.

Phylogeny and host-plant relationships of the Australian Myrtaceae leafmining moth genus Pectinivalva (Lepidoptera, Nepticulidae), with new subgenera and species.

The phylogeny of the mainly Australian nepticulid genus Pectinivalva Scoble, 1983 is investigated on the basis of morphology, and a division into three monophyletic subgenera is proposed on the basis of these results. These subgenera (Pectinivalva, Casanovula Hoare, subgen. n. and Menurella Hoare, subgen. n. ) are described and diagnosed, the described species of Pectinivalva are assigned to them, and representative new species are described in each: Pectinivalva (Pectinivalva) mystaconota Hoare, sp. n., Pectinivalva (Casanovula) brevipalpa Hoare, sp. n., Pectinivalva (Casanovula) minotaurus Hoare, sp. n., Pectinivalva (Menurella) scotodes Hoare, sp. n., Pectinivalva (Menurella) acmenae Hoare, sp. n., Pectinivalva (Menurella) xenadelpha Van Nieukerken & Hoare, sp. n., Pectinivalva (Menurella) quintiniae Hoare & Van Nieukerken, sp. n., and Pectinivalva (Menurella) tribulatrix Van Nieukerken & Hoare, sp. n. Pectinivalva (Menurella) quintiniae (from Quintinia verdonii, Paracryphiaceae) is the first known member of the genus with a host-plant not belonging to Myrtaceae. Pectinivalva (Menurella) xenadelpha from Mt Gunung Lumut, Kalimantan, Borneo, is the first pectinivalvine reported from outside Australia. Keys to the subgenera of Nepticulidae known from Australia, based on adults, male and female genitalia, and larvae, are presented. Host-plant relationships of Pectinivalva are discussed with relation to the phylogeny, and a list of known host-plants of Pectinivalva, including hosts of undescribed species, is presented. DNA barcodes are provided for most of the new and several unnamed species.

Identification of the female-produced sex pheromone of Tischeria ekebladella, an oak leafmining moth.

The native leafmining moth Tischeria ekebladella (Lepidoptera: Tischeriidae) feeds on oaks and recently has become a pest of silviculture and urban green areas in central Europe. The behavioral responses of male moths to hexane extracts of whole bodies of calling females or males were tested under laboratory conditions. Only extracts of females elicited responses from males. Analysis of extracts by coupled gas chromatography/electroantennographic detection revealed the presence of two electroantennogram-active peaks. Structure elucidation of these compounds, by gas-chromatography/mass spectrometry and independent synthesis revealed them to be (3Z,6Z,9Z)-tricosa-3,6,9-triene and (3Z,6Z,9Z,19Z)-tricosa-3,6,9,19-tetraene. While the triene was present in both sexes, the tetraene was female-specific. The latter is a new structure for a pheromone component of Lepidoptera and a novel natural product. Field trapping tests, carried out in a mixed oak forest near Budapest (Hungary), using synthetic compounds applied to rubber dispensers, showed that the tetraene per se elicited catches of males in large numbers. When the triene was added to the tetraene in a ratio of 1:1, there was no increase in trap catch; the triene alone did not elicit catches of males. For monitoring this insect, the tetraene, applied to rubber dispensers at a dose of 300 µg, is a potent sex attractant.

New species in the Old World: Europe as a frontier in biodiversity exploration, a test bed for 21st century taxonomy.

The number of described species on the planet is about 1.9 million, with ca. 17,000 new species described annually, mostly from the tropics. However, taxonomy is usually described as a science in crisis, lacking manpower and funding, a politically acknowledged problem known as the Taxonomic Impediment. Using data from the Fauna Europaea database and the Zoological Record, we show that contrary to general belief, developed and heavily-studied parts of the world are important reservoirs of unknown species. In Europe, new species of multicellular terrestrial and freshwater animals are being discovered and named at an unprecedented rate: since the 1950s, more than 770 new species are on average described each year from Europe, which add to the 125,000 terrestrial and freshwater multicellular species already known in this region. There is no sign of having reached a plateau that would allow for the assessment of the magnitude of European biodiversity. More remarkably, over 60% of these new species are described by non-professional taxonomists. Amateurs are recognized as an essential part of the workforce in ecology and astronomy, but the magnitude of non-professional taxonomist contributions to alpha-taxonomy has not been fully realized until now. Our results stress the importance of developing a system that better supports and guides this formidable workforce, as we seek to overcome the Taxonomic Impediment and speed up the process of describing the planetary biodiversity before it is too late.

Antispila oinophylla new species (Lepidoptera, Heliozelidae), a new North American grapevine leafminer invading Italian vineyards: taxonomy, DNA barcodes and life cycle.

A grapevine leafminer Antispila oinophylla van Nieukerken & Wagner, sp. n., is described both from eastern North America (type locality: Georgia) and as a new important invader in North Italian vineyards (Trentino and Veneto Region) since 2006. The species is closely related to, and previously confused with Antispila ampelopsifoliella Chambers, 1874, a species feeding on Virginia creeper Parthenocissus quinquefolia (L.) Planchon., and both are placed in an informal Antispila ampelopsifoliella group. Wing pattern, genitalia, and DNA barcode data all confirm the conspecificity of native North American populations and Italian populations. COI barcodes differ by only 0-1.23%, indicating that the Italian populations are recently established from eastern North America. The new species feeds on various wild Vitis species in North America, on cultivated Vitis vinifera L. in Italy, and also on Parthenocissus quinquefolia in Italy. North American Antispila feeding on Parthenocissus include at least two other species, one of which is Antispila ampelopsifoliella. Morphology and biology of the new species are contrasted with those of North American Antispila Hübner, 1825 species and European Holocacista rivillei (Stainton, 1855). The source population of the introduction is unknown, but cases with larvae or pupae, attached to imported plants, are a likely possibility. DNA barcodes of the three European grapevine leafminers and those of all examined Heliozelidae are highly diagnostic. North American Vitaceae-feeding Antispila form two species complexes and include several as yet unnamed taxa. The identity of three out of the four previously described North American Vitaceae-feeding species cannot be unequivocally determined without further revision, but these are held to be different from Antispila oinophylla. In Italy the biology of Antispila oinophylla was studied in a vineyard in the Trento Province (Trentino-Alto Adige Region) in 2008 and 2009. Mature larvae overwinter inside their cases, fixed to vine trunks or training stakes. The first generation flies in June. An additional generation occurs from mid-August onwards. The impact of the pest in this vineyard was significant with more than 90% of leaves infested in mid-summer. Since the initial discovery in 2006, the pest spread to several additional Italian provinces, in 2010 the incidence of infestation was locally high in commercial vineyards. Preliminary phylogenetic analyses suggest that Antispila is paraphyletic, and that the Antispila ampelopsifoliella group is related to Coptodisca Walsingham, 1895, Holocacista Walsingham & Durrant, 1909 and Antispilina Hering, 1941, all of which possess reduced wing venation. Vitaceae may be the ancestral hostplant family for modern Heliozelidae.