Deutscher Titel: Josef J. de Freina, Yeray Monasterio León, Carlos Alberto Antonietty & Roger Vila, Anmerkungen zur Biologie, Verbreitung und Systematik von Chondrostega Lederer, 1857, auf der Iberischen Halbinsel mit Beschreibung von Chondrostega escobe- sae sp. nov. aus Südspanien (Lepidoptera: Lasiocampidae, Chondrosteginae) Josef J. de Freina, Yeray Monasterio León, Carlos Alberto Antonietty & Roger Vila, Notes on Chondrostega in the Iberian Peninsula Notes on the biology, distribution and taxonomy of Chondrostega Lederer, 1857 in the Iberian Peninsula with a description of the southern Spanish Chondrostega escobesae sp. nov. (Lepidoptera: Lasiocampidae, Chondrosteginae) ● Josef J. de Freina, Yeray Monasterio León, Carlos Alberto Antonietty & Roger Vila Abstract. Spanish populations of Chon- Resumen. Consideraciones sobre la Zusammenfassung. Von spanischen drostega Lederer, 1857 are examined biología, distribución y taxonomía de Populationen der Gattung Chondrostega based on external and internal morphol- Chondrostega Lederer, 1857 en la pe- Lederer, 1857 werden äußere und inne- ogy as well as molecular data from the nínsula ibérica, con la descripción de re morphologische Merkmale wie auch mitochondrial cytochrome oxidase sub- Chondrostega escobesae sp. nov. del molekulare Datensätze der mitochond- unit I gene (COI). According to relevant sur de España (Lepidoptera: Lasio- rial cytochrome oxidase subunit I gene diagnostic features and genetic differen- campidae, Chondrosteginae) (COI) untersucht und ausgewertet. Die tiation, two species with allopatric dis- Se realiza una revisión taxonómica de las Diagnose belegt die Existenz zweier al- tribution are recognized within the Ibe- poblaciones españolas de Chondrostega lopatrisch wie auch genetisch getrennter rian Peninsula. Andalusian populations, Lederer, 1857, a partir del estudio de Arten auf der Iberischen Halbinsel. Po- here described as Chondrostega escobesae caracteres morfológicos internos y exter- pulationen Andalusiens werden als sp. nov., are compared to the closely re- nos, así como del gen mitocondrial cito- Chondrostega escobesae sp. nov. beschrie- lated Chondrostega vandalicia (Millière, cromo oxidasa subunidad I (COI). Aten- ben und mit ihrer Schwesterart Chond- 1865), which is limited to the northern diendo a características diagnósticas rostega vandalicia (Millière, 1865) ver- half of the Iberian Peninsula. They can relevantes, el género se divide en dos glichen, deren Verbreitung sich auf die be recognized by largely constant mor- especies alopátridas en la península ibé- Nordhälfte der Iberischen Halbinsel be- phological features such as a different rica. Las poblaciones presentes en Anda- schränkt. Beide Arten sind anhand kon- pattern on the underside of the wings of lucía se describen aquí como Chondros- stanter morphologischer Unterschiede the male, adult male genital structure, tega escobesae sp. nov. y son comparadas wie der verschieden ausgeprägten Hin- different structure of the canthus and the con Chondrostega vandalicia (Millière, terflügelzeichnung, der männlichen Ge- colour of hairs in the final instars. Phy- 1865), limitada a la mitad norte de Ibe- nitalstruktur, der Form des Canthus und logenetic analyses based on COI identify ria. Estas especies pueden ser diferencia- der deutlich voneinander abweichenden the two species as sister clades with a das a partir de varias características Färbung erwachsener Raupen unter- minimum genetic divergence of 3.8 %. como la existencia de un patrón diferen- scheidbar. Die Auswertung der DNA- Adults, including the apterous females, te en el reverso de las alas de los machos, Barcoding ergibt einen DNA-Sequenz- immature stages, larval host plants and la estructura genital masculina, diferen- unterschied des mtDNA Cytochromoxy- natural habitat are described and illus- te estructura del canthus y la colouración dase-I-Gens (COI) zwischen nord- und trated. A neotype is designated for C. de la pilosidad de los últimos estadios südspanischen Tieren von mindestens vandalicia. larvarios. El análisis filogenético basado 3,8 %. Der Beitrag liefert Beschreibungen en el COI muestra las dos especies como und Abbildungen der Imagines ein- clados hermanos con una divergencia schließlich des apteren Weibchens, der genética del 3.8 %. Se describen e ilus- letzten Präimaginalstadien, Wirtspflan- tran los adultos de ambos sexos, estadios zen und Lebensräume. Für C. vandalicia inmaduros, plantas nutricias de las oru- wird ein Neotypus designiert. gas y varios hábitats. Se designa además un neotipo para C. vandalicia. Key words. Lepidoptera, Lasiocampoidea, Lasiocampidae, Chondrosteginae, Chondrostega, Iberian Peninsula, new species, neotype, taxonomy, Palaearctic region, DNA barcoding, mitochondrial DNA. Introduction Chondrostega Lederer, 1857 is a complex and western Asia, whereas Chondrostego- dispersed populations. This is especially of species and forms, rather difficult to ides Aurivillius, 1905 is limited to sou- true for the Chondrostega vandalicia (Mil- classify, that is distributed from the sou- thern Africa. Theaptery of Chondrostega lière, 1865) species group, one of the thern Europe and North Africa to central females has presumably resulted in many cases with controversial taxonomy in this 1 Entomologische Zeitschrift · Schwanfeld · 125 (4) 2015 genus (Rougeot & Viette 1978, de Freina the adults. Based on these morphological preserved in absolute ethanol. Total ge- & Witt 1987, Leraut 2006), which dis- differences and mitochondrial DNA re- nomic DNA was extracted using Chelex plays scattered populations from the Ibe- sults, we propose that C. vandalicia exists 100 resin, 100–200 mesh, sodium form rian Peninsula and over the entire Magh- in the northern half of the Iberian Penin- (Biorad), under the following protocol: reb to the Levant. Absence of diagnostic sula, while the Andalusian populations the tissue was introduced into 100 μL of characters both in genitalia and larval represent a separate species, here descri- Chelex 10 % and 5 μL of Proteinase K stages has led to descriptions of a conside- bed as Chondrostega escobesae sp. nov. (20 mg⁄mL) were added. The samples rable number of taxa of uncertain validity, were incubated overnight at 55 °C and listed below: were subsequently incubated at 100 °C for Methods and materials 15 minutes. Samples were then centri- C. constantina Aurivillius, 1894; C. pow- Morphologie and repositories fuged for 10 seconds at 3.000 rpm. A 655- elli Oberthür, 1912; C. tingitana Powell, Cleaned, stained genitalia were stored and bp fragment at the 5’ end of the mitochon- 1916; C. maghrebica de Joannis, 1929 examined in 30 % ethanol, and slide- drial gene (COI) was amplified by poly- (described from populations of Morocco mounted in Euparal before being photo- merase chain reaction (PCR) using the and Algeria); C. zanoni Turati, 1922; C. graphed. primers UniLepF1 (5’-TAA TAC GAC TCA misuratana (Krüger, 1939 (described CTA TAG GGA TTC AAC CAA TCA TAA from Libya); C. subfasciata Klug, 1830; C. Repository abbreviations are as follows: AGA TAT TGG AAC-3’) and UniLepR1 (5’- palaestrana Staudinger, 1891; C. fasciana BC – Barcode [no.]; CFM – Collection de ATT AAC CCT CAC TAA AGT AAA CTT Staudinger, 1891; C. longespinata Auri- Freina, München; CML – Collection Mon- CTG GAT GTC CAA AAA ATC A-3’), with villius, 1894; C. aurivillii Püngeler, 1902; asterio León, Logroño; GPdF – Genitalia universal T7 and T3 tails here highlighted C. goetschmanni Sterz, 1915; C. osthel deri dissection [no.] de Freina; IBEB – Institut in bold.PCR was carried in 25-μL volumes Püngeler, 1925; C. intacta Gaede, 1933; de Biologia Evolutiva (CSIC-UPF), Barce- containing: 14.4 μL autoclaved Milli-Q C. pauli Gaede, 1933; C. schwingenschussi lona; ID – Identifying number; MWM – water, 5 μL 5x buffer, 2 μL 25 mM MgCl2, Zerny, 1933; C. elema Wiltshire, 1941; Collection of Lepidopterological Museum 0.5 μL 10 mM dNTPs, 0.5 μL of each prim- C. brunneicornis Wiltshire, 1944 (all de- Witt, Munich; ZMHU – Museum für er (10 μM), 0.1 μL Taq DNA Polymerase scribed from Levantine populations). Naturkunde, Berlin (formerly Zoologis- (Promega, 5U/μL) and 2 μL of extracted ches Museum der Humoldt-Universität, DNA. The typical thermal cycling profile To test the validity of these taxa, further Germany); ZSM – Bavarian State Collec- was: first denaturation at 92 °C for 60 s, research is essential and molecular data tion of Zoology, Munich. followed by five cycles of 92 °C for 15 s, may shed light on the extent of diversifica- 49 °C for 45 s and 62 °C for 150 s, and then tion among populations. It should be ex- Description and comparative diagnosis is by 35 cycles of 92 °C for 15 s, 52 °C for 45 s pressly mentioned here that due to lack of supplemented by biogeographical and and 62 °C for 150 s and a final extension at scientific verification we can not accept biological data concerning C. vandalicia 62 °C for 420 s. PCR products were purified the arbitrary taxonomic changes within and the new species C. escobesae. This in- and sequenced by Macrogen Inc. using the the Chondrostega group made by Leraut formation has been obtained as a result of T7 and T3 universal primers. Sequences (2006). This means that we still consider intensive observations of immature stages were edited and aligned using GENEIOUS C. vandalicia as non-conspecific with any and life history both in natural conditions PRO 6.0.5 created by Biomatters (http:// of the North African taxa. and captive breeding by Y. M. L., C. A. A. www.geneious.com/). A Neighbour-Join-