Subject Editor: Sven Erlacher
The
Fabricius described
Also
Based on genetic data,
The present study aims to provide a contribution to the clarification of the taxonomic problems still unresolved, through an analysis of the different populations that make up this complex.
The study is based on material deposited in the collections listed in Table
Abbreviations of the collections.
Taxon delimitation was based on the combined study of external morphology of the adults (coloration and pattern of the upperside and underside wings), morphology of the female genitalia (signum and lamella postvaginalis) and tympanal organs, as well as of molecular data.
For the extraction of the female genitalia and tympanal organs the abdomen was macerated cold in a 20% KOH solution for 12 hours. Then the longitudinally sectioned abdomen was stained using chlorazol black-alcohol mixture for two minutes. Genitalia were cleaned in water and mounted on a slide in dimethyl hydantoin formaldehyde (DMHF). Tympanal organs were removed from the abdomen, cleaned and mounted on a slide.
Because the signum is stellate with a more or less elliptical shape, in each specimen the smaller diameter and the larger diameter of this ellipse were measured (both up to the ends of the tips), averaging the two diameters.
For the DNA analyses, one leg was removed from dried specimens. DNA extraction, amplification, and sequencing of the barcode region of the mitochondrial cytochrome c oxidase I (COI) gene (658 base pairs) were carried out in the Canadian Centre for DNA Barcoding, Ontario, Canada, using standard protocols (
The Barcode Index Number (BIN) is an online framework of BOLD database that clusters barcode sequences algorithmically (
Information on molecular data was provided by A. Hausmann (SNSB-Bavarian State Collection of Zoology, Munich, Germany), who also examined the specimens from the ZSM collection. Sequence divergence was calculated by Hausmann using the Kimura 2-parameter model (
The results from the molecular analysis are shown in Figure
The
Figure
Barcoded specimens (from BOLD data systems).
Taxon | Country | Region | Locality and date | Collector and collection | Specimen ID | Barcode Index Number (BIN) |
---|---|---|---|---|---|---|
Germany | Hesse | Lahn-Dill-Kreis, Hirzenhain, 4.vii.2002 | D. Feierabend leg., |
BC ZFMK Lep 00683 |
|
|
Germany | Thuringia | Gumpelstadt, Jauchsental, 11-vi.2006 | U. Buechner leg., |
SE |
|
|
Germany | Thuringia | Wahlhausen, 22.v.1998 | R. Rommel leg., |
SE |
|
|
Germany | Bavaria | Oberpfalz, Schwarzenbach, 11.v.1994 | G. Nowak leg., |
SE |
|
|
France | Upper Normandy | Seine Maritime, Anneville-Ambourville, 22.v.1995 | B. Dardenne leg., |
LN-BD1103 |
|
|
|
Italy | Liguria | Ligurian Alps, Ventimiglia, 600 m, 27.v.2001 | E.O. Bonora leg., |
BC ZFMK Lep 00684 |
|
|
Italy | Tuscany | Apuan Alps, under Monte Corchia, Passo Croce, 18.vi.1999 | L. Dapporto leg., ZSM | BC ZSM Lep 63447 |
|
|
Italy | Lombardy | Bergamasque Prealps, Camerata Cornello, towards Passo Grialeggio, 23.vii.2010 | M. Massaro and W. Zucchelli leg., |
BC ZSM Lep 97046 |
|
|
Italy | Trentino-Alto Adige | Monte Baldo, 1550 m, 8.vi.1994 | S. Erlacher leg., |
SE |
|
|
Austria | Carinthia | [Gitschtal], |
Ch. Wieser leg., |
|
|
|
Italy | Friuli-Venezia Giulia | Julian Prealps, Val Venzonassa, Jof Ungarina SW side – Malga Confin W, 1280 m, 12.vi.2007 | P. Huemer leg., |
|
|
|
Bulgaria | W Stara Planina Mts | between Gorni Lom Village and Midzhur Top, 1425 m, 14.vii.2009 | S. Beshkov leg., |
BC SB Lep 0017 |
|
|
Romania | Buzău County | Eastern Carpathians, Nemira Mountains, Lassuag, 1100 m, 22.vi.1996 | S. and.Z. Kovacs leg., ZSM | BC ZSM Lep 73989 |
|
|
Romania | Dâmbovița County | Southern Carpathians, Bucegi Mountains, Valea Jepii, 1800 m, 7.vi.2007 | S.and.Z. Kovacs leg., ZSM | BC ZSM Lep 73986, BC ZSM Lep 73987 |
|
Germany | Thuringia | Erlau, Zeltplatz, 4.vii.1999 | S. Erlacher leg., |
SE |
|
|
Germany | Saxony-Anhalt | Bitterfeld, Marke: 0.5 km east the motorway exit Dessau-Süd, 85 m, 24.v.2012 | J. Gelbrecht leg., ZSM | BC ZSM Lep 91967 |
|
|
Germany | Bavaria | Spalt, 6.v.1997 | A. Zoglauer leg., |
BC ZFMK Lep 00685 |
|
|
Germany | Bavaria | Mittelfranken, Roth, Spalt, 400 m, 17.vii.2007 | A. Zoglauer leg., |
BC ZSM Lep 24010 |
|
|
Germany | Bavaria | Mittelfranken, Nuernberger Land, Leinburg, Wolfsgrube Ost 405 m, 20.vii.2010 | A. H. Segerer leg., ZSM | BC ZSM Lep 51392 |
|
Distribution of the BINs of the complex
Distribution of the taxa of the complex
The diagnostic characters of
According to
J. Gelbrecht (pers. comm.) found differences in the size of the signum in the female genitalia.
The morphological characters observed in the material examined by us are summarized in Table
Our observations confirm that the posterior margin of the lamella postvaginalis is often more curved in
Upperside of males (3–7) and females (8–12).
Underside of males (13–17).
Underside of females (18–22).
Variability of the lamella postvaginalis.
Variability of the bullae tympani (bars point at the rounded lobes).
J. Gelbrecht (pers. comm.) has bred material of the taxa
Gelbrecht also did breeding experiments (pers. comm.). The taxa
We have not found any images of the preimaginal stages of the Italian populations and we do not believe that breeding experiments have ever been carried out between them.
Populations delineated on the basis of molecular data, the size of the signum and the presence (or absence) of a roundish lobe in the bullae tympani, can be divided into two groups: one including the taxa
Furthermore, the two groups of populations are not completely separated from each other. Some populations of the northern Apennines (taxon
The genetic distance between the two groups of populations of the
Gelbrecht’s breeding experiments suggest the conspecificity between
These results emphasize the need to further investigate the patterns of mitochondrial DNA in “hybrid” populations. In our research on
Analysis of the morphological characters observed in the material examined.
Taxon | Upperside | Underside | Female genitalia and tympanal organs |
---|---|---|---|
BIN | |||
place of origin of the material | |||
(♂♀) Forewing more or less irrorated with brown scales (sometimes almost absent except in the costa and in the terminal fascia). Width of the forewing dark terminal fascia about 2 mm (rarely 1 mm), hindwing dark terminal fascia generally well developed, rarely reduced; hindwing often densely covered with dark scales. | (♂♀) Forewing scattered with dark vertical stripes or more densely sprinkled with dark scales. Hind wing more or less densely covered with dark scales, except the whitish radiating streaks. | Signum: 0.7–1 mm (n=2, from Spain and Belgium). | |
|
Bullae tympani with a roundish lobe (visible only in one bulla in a female from Spain). | ||
Spain, Belgium, Germany (North Rhine-Westphalia), Switzerland | |||
|
(♂♀) Forewing almost free of dark scales (except the costa and the terminal fascia). Hindwing with more dark scales. Width of the forewing dark terminal fascia generally just over 1 mm (rarely 2 mm, 3 mm in the |
(♂♀) Hindwing scattered with dark vertical stripes (much less numerous on the forewing). Whitish radiating streaks of the hindwing present and generally well evident (barely perceptible in the |
Signum: 0.8–0.9 mm (exceptionally 1.2 mm in a female from the Apennines near Bologna; 0.9 mm in another female from the same locality) (n=5, from France and Italy: Alpes-Maritimes, Ligurian Alps, Ligurian Apennines and Tuscan-Emilian Apennines). |
|
Bullae tympani with a roundish lobe (France SE, Ligurian Alps, Ligurian Apennines); a female from the Tuscan-Emilian Apennines (near Bologna) lacks it, in a male from the same locality it is barely visible. | ||
France SE, Italy (Ligurian Alps, Northern Apennines) | |||
|
(♂♀) Forewing almost free of dark scales (except the costa and the terminal fascia). Hindwing with a few dark scales. Width of the forewing dark terminal fascia 1.5–2 mm, hindwing dark terminal fascia narrow or reduced to a few scales. | (♂♀) Hindwing scattered with dark vertical stripes (much less numerous on the fore wing). Whitish radiating streaks of the hindwing present. | Signum: 1.4 mm (n=1). |
not barcoded | Bullae tympani without roundish lobe. | ||
Italy (northern Piedmont: Macugnaga) | |||
|
(♂♀) Forewing often with only a few dark scales (except the costa and the terminal fascia), a little more dense in the hindwing; sometimes both wings scattered with dense groups (or short stripes) of dark scales. Width of the dark terminal fascia generally just over 1 mm. | (♂♀) Fore- and hindwing scattered with dark vertical stripes; ground colour of the wings almost identical or more orange in the fore wing. Whitish radiating streaks of the hindwing present. | (altogether in the taxon |
|
Signum: 1.1 mm (exceptionally 1.6 mm in a female from Lombardy) (n=6, from northeastern Piedmont, Lombardy, Monte Baldo, Carnic Prealps, Italian Julian Alps and Bulgaria). | ||
Italy (northeastern Piedmont, Lombardy, Monte Baldo) | Bullae tympani without roundish lobe. | ||
|
(♂♀) Wings scattered with sparse groups (or short stripes) of dark scales, sometimes with more dense dark vertical stripes. Width of the dark terminal fascia just over 1 mm. | (♂♀) Fore- and hindwing scattered with dark vertical stripes (more dense on the hindwings); ground colour of forewing more orange than the hindwing. Whitish radiating streaks of the hindwing present. | |
|
|||
Julian Alps (Italy and Slovenia) | |||
|
(♂) Forewing scattered with sparse groups (or short stripes) of dark scales, hindwing with dense dark vertical stripes. Width of the dark terminal fascia just over 1 mm. | (♂) Fore and hindwing scattered with dense dark vertical stripes; ground colour of forewing more orange. Whitish radiating streak of the hindwing present. | |
not barcoded | |||
(most likely |
|||
Macedonia | |||
|
(♂♀) Ground colour yellowish scattered with irregular dark vertical stripes and groups of dark scales, often more dense in the hindwing. Width of the dark terminal fascia just under 2 mm. | (♂♀) Fore- and hindwing scattered with dark vertical stripes; ground colour of fore wing slightly more orange. Whitish radiating streak(s) of the hindwing present but generally not very evident (sometimes barely perceptible). | (altogether in the taxon |
|
Signum: 1.1 mm (exceptionally 1.6 mm in a female from Lombardy) (n=6, from northeastern Piedmont, Lombardy, Monte Baldo, Carnic Prealps, Italian Julian Alps and Bulgaria). | ||
Bulgaria (Vitosha) | Bullae tympani without roundish lobe. | ||
|
(♂♀) Very similar to the Bulgarian specimens. | (♂♀) Very similar to the Bulgarian specimens. | |
|
|||
Romania (Buzău and Dâmbovița Counties) | |||
(♂♀) Ground colour deep yellow with irregular dark vertical stripes, sometimes divided into groups of scales (especially in the forewing) or thickened one to the other. Width of the dark terminal fascia 2–3 mm. | (♂♀) Fore- and hindwing with identical pattern: ground colour orange yellow scattered with dark vertical stripes (a little more dense on the hind wings). Hindwing without a trace of whitish radiating streaks. | (altogether in the taxon |
|
|
Signum: 1.2–1.4 mm (n=3, from Slovakia and Germany). | ||
Czech Republic, Slovakia | Bullae tympani without roundish lobe. | ||
(♂♀) Fore- and hindwing with identical pattern: ground colour orange scattered with dark vertical stripes. Width of the dark terminal fascia 2–2.5 mm, rarely less in very small specimens. | (♂♀) Fore- and hindwing with identical pattern: ground colour orange scattered with dark vertical stripes (a little more dense on the hind wings). Hindwing without a trace of whitish radiating streaks. | ||
|
|||
Germany (Bavaria) |
Variability of the signum.
Pupal cremaster (drawings by J. Gelbrecht).
Our findings indicate that the relation of barcodes to morphology is a complicated issue, and a ‘correct’ taxonomic interpretation may be impossible to reach solely with these approaches. The case of
Colour images of specimens from Great Britain are shown in
(Figs
In the female genitalia signum small (0.7–1 mm). Bullae tympani with a roundish lobe (visible only in one bulla in a female from Spain).
Great Britain (Scotland and eastern England, now extinct), part of Germany (cf.
According to
(Figs
In the female genitalia signum small (0.8–0.9 mm, exceptionally 1.2 mm in a female from the Apennines near Bologna: see Table
France SE, Italy (Piedmont, Liguria, Tuscany, Emilia-Romagna: see Fig.
The two barcoded specimens (both from Italy) diverge from those of the taxon
The distribution of the taxa
(Figs
In the only female examined signum large (1.4 mm). Bullae tympani without roundish lobe.
Valle Anzasca (Italy: northern Piedmont), on the southern side of the Monte Rosa massif (Fig.
Not yet barcoded. The size of the signum and the absence of a roundish lobe in the bullae tympani indicate a greater similarity with
The underside of a male from Gitschtal (Austria: Carinthia) is figured by
(Figs
In the female genitalia signum large (1.1 mm, exceptionally 1.6 mm). Bullae tympani without roundish lobe.
Northern Italy (see Fig.
The westernmost populations (Lombardy and Monte Baldo in Italy, Carinthia in Austria) correspond to a separate BIN, at a distance of 2.0%, while all the others (from the Julians Alps to Bulgaria and Romania) share the same BIN of the following subspecies; however, no constant morphological character corresponds to these genetic differences and it is not possible to distinguish the specimens of the more western regions from those of the Julian Alps (both populations are very variable). The exact border between these two BINs is not known, since no specimens from the Carnic Prealps have been barcoded. Three specimens from Romania diverge into a separate cluster, but they are morphologically very similar to those from Bulgaria.
Colour images of specimens from Poland are shown by
(Figs
On the upperside, the dark terminal fascia is variable: according to
In the female genitalia signum large (1.2–1.4 mm). Bullae tympani without roundish lobe.
North-eastern and south-eastern Germany (cf.
We are grateful to Axel Hausmann (SNSB-Bavarian State Collection of Zoology, Munich, Germany), who examined the specimens from the
Neighbour joining tree (Kimura 2-parameter distance model for COI-5P marker, data from BOLD data systems) for 23 European specimens. Terminals with taxon, specimen ID and geography.