Corresponding author: Peter Huemer (
Academic editor: A. Zilli
We explore the potential value of DNA barcode divergence for species delimitation in the genus
Huemer P, Karsholt O, Mutanen M (2014) DNA barcoding as a screening tool for cryptic diversity: an example from
The genus
Extensive generic descriptions and diagnoses of European species of
Specimens. Our study is based on about 50 specimens of the
DNA Barcodes. Full-length lepidopteran DNA barcode sequences are a 648 base-pair long segment of the 5’ terminus of the mitochondrial COI gene (
Photographic documentation. Photographs of the adults were taken with an Olympus SZX 10 binocular microscope and an Olympus E 3 digital camera and processed using the software Helicon Focus 4.3 and Adobe Photoshop CS4 and Lightroom 2.3. Genitalia photographs were taken with an Olympus E1 Digital Camera from Olympus BH2 microscope.
The Natural History Museum (British Museum, Natural History) London (United Kingdom)
Tiroler Landesmuseum Ferdinandeum, Innsbruck, Austria
Zoological Museum, University of Helsinki, Finland
Zoological Museum, Natural History Museum of Denmark, Copenhagen, Denmark
Zoological Museum, University of Oulu, Finland
Forty-four of 51 European species were successfully sequenced, resulting in a full-length barcode fragment for 191 specimens and more than 500 bp for further 26 specimens (
Neighbour-joining tree (Kimura 2 parameter, built with MEGA 5; cf.
Intraspecific mean K2P (Kimura 2 Parameter) divergences, maximum pairwise distances and distance to nearest neighbor.
Species | Mean Intra-Sp | Max Intra-Sp | Nearest Neighbour | Nearest Species | Distance to NN |
---|---|---|---|---|---|
|
4.85 | 4.85 | PHLAE427-11 |
|
3.81 |
|
3.05 | 4.76 | LEATC402-13 |
|
5.21 |
|
0.16 | 0.3 | LEFIK150-10 |
|
5.78 |
|
0.4 | 0.81 | LEFIB755-10 |
|
5.78 |
|
0.21 | 0.46 | LEATD656-13 |
|
3.94 |
|
0.42 | 0.61 | PHLAI019-12 |
|
5.07 |
|
1.99 | 6.95 | PHLAA069-09 |
|
6.61 |
|
N/A | N/A | PHLAF489-11 |
|
4.54 |
|
0.21 | 0.31 | LEATC-402-13 |
|
5.41 |
|
0 | 0 | PHLAB900-10 |
|
5.29 |
|
1.02 | 1.39 | PHLAI203-13 |
|
4.57 |
|
3.4 | 6.27 | LEFIF467-10 |
|
4.67 |
|
0 | 0 | LEFIC281-10 |
|
4.5 |
|
0.47 | 1.7 | PHLAI156-12 |
|
4.55 |
|
N/A | N/A | PHLAI019-12 |
|
6.54 |
|
N/A | N/A | LEFIJ1014-11 |
|
4.88 |
|
0.4 | 0.77 | PHLAI156-12 |
|
5.55 |
|
1.08 | 1.08 | PHLAI019-12 |
|
4.39 |
|
0 | 0 | LEFIF480-10 |
|
4.55 |
|
4.25 | 4.25 | PHLAA055-09 |
|
5.56 |
|
0.31 | 0.61 | LEEUA184-11 |
|
4.9 |
|
N/A | N/A | PHLAI447-13 |
|
5.29 |
|
1.47 | 3.79 | PHLAG331-12 |
|
3.76 |
|
0.12 | 0.3 | PHLAG331-12 |
|
4.24 |
|
0 | 0 | LEEUA182-11 |
|
0.3 |
|
1 | 1.54 | PHLAI203-13 |
|
0.3 |
|
N/A | N/A | LEATE421-13 |
|
3.76 |
|
0.3 | 0.46 | PHLAE427-11 |
|
4.87 |
|
N/A | N/A | LEEUA388-11 |
|
3.81 |
|
3.58 | 5.69 | PHLAB899-10 |
|
3.93 |
|
0.97 | 2.26 | PHLAH147-12 |
|
4.88 |
|
0.23 | 0.61 | PHLAD576-11 |
|
3.85 |
|
0.41 | 0.61 | PHLAI019-12 |
|
3.94 |
|
2.07 | 3.61 | LEATC292-13 |
|
3.12 |
|
0 | 0 | PHLAE429-11 |
|
3.33 |
|
0 | 0 | LEFIJ778-10 |
|
6.61 |
|
N/A | N/A | PHLAD573-11 |
|
3.42 |
|
0.06 | 0.15 | PHLSA085-11 |
|
3.42 |
|
0.77 | 1.24 | PHLSA085-11 |
|
3.74 |
|
N/A | N/A | PHLAD576-11 |
|
3.33 |
|
N/A | N/A | PHLAG580-12 |
|
4.54 |
|
1.09 | 2.02 | PHLAD576-11 |
|
4.01 |
|
0 | 0 | PHLAB622-10 |
|
3.93 |
|
0 | 0 | PHLAI014-12 |
|
4.67 |
|
1.48 | 2.7 | PHLAF105-11 |
|
5.36 |
|
0.22 | 0.47 | LEEUA388-11 |
|
4.16 |
Sequences of the COI barcode region of all analysed morphospecies reveal significant interspecific genetic distances with barcode gaps ranging from a minimum of 3.11% to the nearest neighbour (
The
Adults.
Male genitalia.
Adult (
Variation. No variation observed except for size, which differs considerably in two reared specimens from Italy and Greece.
The name “
The species is known from widely separated localities in northern Italy, Switzerland and Greece, indicating a more widespread distribution in Sub-Mediterranean and Mediterranean Europe. However, the host-plants are much more widespread, ranging to northern Europe in the north and to Central Asia in the east. No sympatric occurrence with
The larva has been found in early spring, feeding in the stem of
The majority of collected material belongs to females whereas
Lectotype ♂[with nine labels]: ‘Fennia Ab Bromarf’ ‘R. F:tius’ ‘21.7.21’ [piece of celluloid where genitalia was mounted] ‘Type ♂’ [red] ‘Lita amaurella m. det. Mart. Hering ♂’ ‘Mus. Zool. H:fors spec. typ. No 7016 Lita amaurella Hering’ ‘Lita amaurella m. ♂ Sch.-Armatur Bromarf 21.7.21 Fabritius’ ‘LECTOTYPE O. Karsholt design.’.
See above.
Adult (
Variation. The colour of the forewings varies from greyish to blackish. Worn specimens look lighter than fresh ones. Sometimes there are no white scales in the middle of the wing.
Male genitalia.
Details of male genitalia (vinculum-valva-complex).
Female genitalia.
Female genitalia.
With certainty known from scattered records from northern and Central Europe and Turkey. All the specimens from north of the Alps that we have been able to cross-check are correctly attributed to
Turkish specimens of
One of the examined specimens of
The genus
Intraspecific variation shows a different pattern. The majority of species has a low (<2%) maximum intraspecific divergence and thus seems taxonomically well defined. However, a remarkable number of species (8 species, nearly one quarter of all, 9 species with only one sample not considered) is characterized by maximum divergence exceeding 3% (
We are particularly grateful to Paul Hebert and his team at the Canadian Centre for DNA Barcoding (Guelph, Canada), whose sequencing work was enabled by funding from the Government of Canada to Genome Canada through the Ontario Genomics Institute. We are also grateful to the Ontario Ministry of Research and Innovation and to NSERC for their support of the BOLD informatics platform.
Stefan Heim (TLMF) is acknowledged for his kind assistance with photographic work. We thank Robert J. Heckford (Plymouth, GB), Petri Hirvonen (Porvoo, Finland), Thierry Varenne (Nice, France), Christian Wieser (Klagenfurt, Austria) and Josef Wimmer (Steyr, Austria) for providing material for our examination, and Lauri Kaila (ZMUH) for access to the type of
We are particularly indebted to the Promotion of Educational Policies, University and Research Department of the Autonomous Province of Bolzano - South Tyrol for helping to fund the project “Genetic biodiversity archive - DNA barcoding of
Last, but not least, we thank Martin Corley (Faringdon, UK) for linguistic improvement of the manuscript.
Sample information for specimens included in this study.
Data type: species data
Explanation note: Process IDs are sequence identifiers in BOLD; Sample IDs are specimen identifiers; BINs are Barcode Identification Numbers in BOLD. Details of collecting data, images, sequences, and trace files for the barcoded specimens are available in the public BOLD dataset “DS-LECARY”, accessed at