Tijdschrift voor Entomologie 156 (2013) 191–210 brill.com/tve The Bupleurum (Apiaceae) feeding species of Trifurcula (Glaucolepis): new species, biology and distribution (Lepidoptera: Nepticulidae) Zdenekˇ Laštuvka,˚ Aleš Laštuvka˚ & Erik J. van Nieukerken The Bupleurum feeding species of Trifurcula (Glaucolepis) Braun, 1917 are revised. Five species are recognised: T. bupleurella (Chrétien, 1907), T. sanctibenedicti Klimesch, 1979, T. megaphallus van Nieukerken, Z. Laštuvka˚ & A. Laštuvka˚ sp. n. feeding on Bupleurum gibraltarium in southern Spain, T. chretieni Z. Laštuvka,˚ A. Laštuvka˚ & van Nieukerken sp. n. feeding on Bupleurum rigidum in southern France, Spain and Portugal, and T. siciliae Z. Laštuvka,˚ A. Laštuvka˚ & van Nieukerken sp. n. feeding on B. fruticosum in Sicily. The group is restricted to southwestern Europe and northern Africa, the area where most shrubby Bupleurum species occur. A NJ and Bayesian analysis of DNA barcodes of four out of five species suggest a single origin of Bupleurum feeding in the subgenus Glaucolepis. Zdenekˇ Laštuvka*,˚ Mendel University, Zemedelská 1, CZ-613 00 Brno, Czech Republic. [email protected] Aleš Laštuvka,˚ Slavíckovaˇ 15, Prostejov,ˇ Czech Republic Erik J. van Nieukerken, Department of Terrestrial Zoology, Naturalis Biodiversity Center, PO Box 9517, NL-2300 RA Leiden, The Netherlands. [email protected] Introduction set of the (sub)genus (van Nieukerken & Johansson The subgenus Glaucolepis Braun, 1917 of the genus 1990, Klimesch 1977, Laštuvka˚ & Laštuvka˚ 1997, Trifurcula Zeller, 1848 is diverse in the Mediter- van Nieukerken et al. 2010). Hitherto two species ranean region, where the majority of the 30 named were known to feed on the genus Bupleurum,a species is found. Whereas the type species, T. (Glau- genus of Apiaceae with several shrubby species in the colepis) saccharella (Braun, 1912), is the only known Mediterranean. Despite the fact that Apiaceae usu- North American species of the genus, and a leafminer ally harbour large numbers of leafmines (most fre- on Acer, the Mediterranean species make leaf- or quently caused by Diptera: Agromyzidae, eg. Ellis stem-mines on small shrubs and herbs belonging 2012), they are a very unusual hostplant family for to Lamiaceae, Plantaginaceae (genus Globularia), Nepticulidae. The only other known nepticulid to Apiaceae (genus Bupleurum) and single species on feed on these plants is the taxonomically isolated and Linaceae (Linum) and Asteraceae (Launaea)respec- rare European Ectoedemia (Fomoria) viridissimella tively. A revision for the subgenus as a whole does (Caradja, 1920) that feeds on Peucedanum cervaria not yet exist, but several recent papers treat a number (L.) Lapeyr. (van Nieukerken & Johansson 1990, of species taxonomically (Klimesch 1975, 1976, van Laštuvka˚ & Laštuvka˚ 1997). The known Bupleurum Nieukerken & Puplesis 1991, Laštuvka˚ & Laštuvka˚ feeders are T. bupleurella (Chrétien 1907), reared 1998, 2007, Laštuvka˚ & Laštuvka˚ 2000, Ivinskis et from Bupleurum fruticosum and T. sanctibenedicti al. 2012) and some papers review at least a sub- Klimesch 1979 from B. fruticescens. During our field- Tijdschrift voor Entomologie 156: 191–210, Table 1. Figs 1–51. [ISSN 0040-7496]. brill.com/tve © Nederlandse Entomologische Vereniging. Published by Koninklijke Brill NV, Leiden. Published 20 December 2013. DOI 10.1163/22119434-00002028 Downloaded from Brill.com10/11/2021 07:29:54AM * Corresponding author via free access 192 Tijdschrift voor Entomologie, volume 156, 2013 work in the western Mediterranean, we encountered DNA barcoding leafmines on several other Bupleurum species, partly DNA was extracted from caterpillars or from dry belonging to new taxa. We describe here three new adult abdomens. DNA extraction from larvae was species and provide new data for the known species, usually destructive; from abdomens and some lar- including DNA barcoding data for four out of the vae the non-destructive protocol by Knölke et al. five species. (2005) was followed, allowing the preparation of the genitalia or larval skin as well. Details of meth- ods are presented by van Nieukerken et al. (2012), Material and methods weprovideheretheCOIDNAbarcodeforfour In the material lists we do not repeat records from out of the five species, only lacking Trifurcula si- France, Spain or Portugal that we published earlier ciliae sp. n. They are compared with and included (Laštuvka˚ & Laštuvka˚ 2011, van Nieukerken et al. in the recently published dataset of Glaucolepis bar- 2004, 2006). Adults were collected at various types codes (Ivinskis et al. 2012), with Trifurcula (s.s.) pal- of light (8 to 25 W fluorescent tubes, UV lamp lidella (Duponchel, 1843), type species of the genus, 125 W) or during the day by beating the host plants as outgroup. Data of all barcodes and vouchers can with a sweep net; mines with larvae were collected be found on the Barcode of Life webpages (http:// and partly reared, or larvae taken out and kept in www.barcodinglife.com/views/login.php) under the ethanol for morphological or molecular analysis. The dataset “Bupleurum feeding Trifurcula (Glaucolepis)” genitalia of both sexes were prepared in the usual [doi: dx.doi.org/10.5883/DS-BUPGLAU]. We give way (e.g. van Nieukerken 1985, van Nieukerken et here the specimen Registration numbers, and the al. 1990), genitalia were either embedded in perma- Barcode Identification Number (BIN) for each nent slides, using euparal, or in small vials with glyc- species, and Genbank accession numbers when avail- erine. Watercolours and drawings were prepared by able. For the most recent records they were not yet AL, those of genitalia were often made from genitalia available when going to press and can later be found embedded in glycerine, which could be easier manip- in the BOLD dataset. ulated and then viewed from all sides. Photographs of moths, leafmines and genitalia slides were taken Analysis by EvN with a Zeiss AxioCam (HR or MR5) dig- Neighbor Joining (NJ) trees based on DNA barcode ital camera attached respectively to a Zeiss Stemi sequences of all available specimens were prepared SV11 stereo-microscope and a Zeiss Axioskop H, us- with tools on BOLD (Ratnasingham & Hebert ing Carl Zeiss AxioVision software (version 4), for 2007). Genetic distance calculations were performed some photographs using the module “Extended fo- using uncorrected P distance (Srivathsan & Meier cus”. Hostplant names follow Hand (2011). Dis- 2012). tribution maps were prepared with Dmap (Morton A Bayesian Analysis was carried out with the same 2000). dataset in MrBayes 3.1.2 (Ronquist & Huelsenbeck For the sake of comparability the morphological 2003). Each analysis was run twice, starting from terms follow our earlier treatments of the genus random starting trees, for 1 million generations and (e.g. Nieukerken & Johansson 1990, Laštuvka˚ & sampling from the posterior every 1000 generations. Laštuvka˚ 2007, van Ivinskis et al. 2012), even though No partition was used. A conservative burn-in of the correct morphological term for aedeagus would 25% was chosen. be phallus and terminal cilia and cilia-line could better be termed fringe and fringe-line. Taxonomy Material is either from the collection of A. Laštuvka˚ (AL), or from one of the following museum All species and hostplants are listed in Table 1. collections: Musée Nationale de l’Histoire Naturelle, Paris (France), Naturalis Biodiversity Center, Lei- Trifurcula (Glaucolepis) bupleurella den (RMNH), Staatliches Museum für Naturkunde (Chrétien) Stuttgart (Germany), Zoologische Staatssammlung Figs 1, 10, 15, 16, 20, 25, 26, 31, 32, 46 München (Germany), Zoological Museum, Natural History Museum of Denmark, Copenhagen (Den- Nepticula bupleurella Chrétien 1907: 91. Syntypes (3): [France], Hérault, Aude, mines on Bupleurum fruticosum, mark). We refer to the collections by their cities. The September, adults emerged June, July, Chrétien (MNHN) code RMNH is used for registry numbers in Lei- [not examined]. den. Ectoedemia (Dechtiria) bupleurella: Klimesch 1975: 863 [re- description]. Trifurcula (Glaucolepis) bupleurella: van Nieukerken 1986: 15 [new combination]; Laštuvka˚ & Laštuvka˚ 1997: 131 [re- Downloaded from Brill.com10/11/2021 07:29:54AM via free access Laštuvka˚ et al.: Bupleurum feeding Trifurcula (Glaucolepis) 193 Table 1. List of species and hostplants. Trifurcula (Glaucolepis) bupleurella (Chrétien, 1907) Bupleurum fruticosum L. ∗ Trifurcula (Glaucolepis) megaphallus sp. n. Bupleurum gibraltarium Lam. Trifurcula (Glaucolepis) chretieni sp. n. Bupleurum rigidum L. subsp. rigidum Bupleurum rigidum subsp. paniculatum (Brot.) H. Wolff Trifurcula (Glaucolepis) siciliae sp. n. Bupleurum fruticosum L. Trifurcula (Glaucolepis) sanctibenedicti Klimesch, 1979 Bupleurum fruticescens L. subsp. fruticescens Bupleurum fruticescens subsp. spinosum (Gouan) O. Bolòs & Vigo ∗ Often incorrectly spelled as gibraltaricum. description]; van Nieukerken et al. 2004: 231 [distribution Female genitalia (Figs 20, 25, 26). Total bursa length Spain, Portugal]; van Nieukerken et al. 2006: 62 [distribu- 835–950 μm. Anal papillae distinct, rounded, with tion France]. ca. 35 short setae each. T8 with groups of short setae and scales; S8 with two transversal rows of several Diagnosis long setae. Posterior and anterior apophyses of same Recognised from other Bupleurum feeders by the length, thin. Signa about 310–365 μm, up to 6– combination of its size, large tornal spot and in male 8rowsofcellswide,threetimesaslongaswide. the absence of yellow androconial scales on forewing