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NoTA LEPIDOPTEROLOGICA

A journal focussed on Palaearctic and General Lepidopterology

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Cover image: Ochromolopis ictella, imago (Fig. 1 in Gaedike & Mally: On the taxonomic status of Ochromolopis

ictella (Hübner, 1813) and O. zagulajevi Budashkin & Sachkov, 1991 (Lepidoptera, Epermeniidae))

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NOTA LEPIDOPTEROLOGICA

VOLUME 37 No. 1 : Sofia, 15.06.2014 ° ISSN 0542-7536

Jadranka Rota. Nota Lepidopterologica combines tradition and

innovation through open access and advanced publishing model....... 1-2

Vladimir Hula, Jana Niedobova. The first record of Pyrgus

malvoides (Elwes & Edwards, 1897) in the Czech Republic

(Lepidoptera, Hesperiidae): an accidental introduction? .................... 3-8

Balazs Toth, Laszlo Ronkay. Revision of the Palaearctic and

Oriental species of the genus Naarda Walker, 1866 (Lepidoptera,

Erebidae, Hypeninae). Part 3. Description of three new species

IROTENSI A ee ee re 9-18

Frans Cupedo. Reproductive isolation and intraspecific structure in

Alpine populations of Erebia euryale (Esper, 1805) (Lepidoptera,

Neanphalidae, Saba) De den di nou 19-36

Hossein Rajaei Sh., Gyula M. Laszlo. Scotopteryx kurmanjiana,

a new species from the Kopet-Dagh Mountains (Lepidoptera,

Geomeinigae, Lärenlinae) sen ae 37-42

Bernard Landry, Giorgio Baldizzone. Description of the reduced

mouth parts of Coleophora micronotella Toll (Lepidoptera,

Coleophoridae), with a New SYNONVML 1.104 43-48

Reinhard Gaedike, Richard Mally. On the taxonomic status

of Ochromolopis ictella (Hübner, 1813) and O. zagulajevi

Budashkin & Sachkov, 1991 (Lepidoptera, Epermeniidae)............ 49-62

Dmitry F. Shovkoon, Tatiana A. Trofimova. Description of the

female of Ethmia cribravia Wang and Li 2004 (Lepidoptera,

Fac las ticle ple ANA), ee ten 63-65

Oleksiy V. Bidzilya. A remarkable new species of the genus

Catatinagma Rebel, 1903 (Lepidoptera, Gelechiidae) from

Turkmenistan... au ee ee ene eee eee 67-74

Jure Jugovic, Toni Koren. Wing pattern morphology of three

closely related Melitaea (Lepidoptera, Nymphalidae) species

reveals highly inaccurate external morphology-based species

IJENLIRCAUION.. na ee SERRE 75-90

Jadranka Rota, Antonio M. F. Aguiar, Ole Karsholt. Choreutidae of

Madeira: review of the known species and description of the male

of Anthophila threnodes (Walsingham, 1910) (Lepidoptera)....... 91-103

Book review: I Lepidotteri del Parco Naturale delle Capanne di

MTarCarolo..... ere nese eee 105-106

Nota Lepi. 37(1) 2014: 1-2 | DOI 10.3897/n1.37.8008

Nota Lepidopterologica combines tradition and innovation through

open access and advanced publishing model

JADRANKA Rota!

1 Laboratory of Genetics/Zoological Museum, Department of Biology, University of Turku, FI-20014 Turku,

Finland; jadranka.rota@utu.fi

The Societas Europaea Lepidopterologica (SEL) has joined forces with Pensoft as our new

publisher to bring the Society’s journal Nota Lepidopterologica on the way to open access and

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The scope of Nota has not changed and we continue to publish contributions to the study of

mainly but not exclusively Palaearctic Lepidoptera, including taxonomy, morphology and anat-

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2 Editorial

per year, one on June 15 and one on December 15, in which we will gather all the papers that

were published online in the previous six months.

We hope you will enjoy the new editorial system and outlook of Nota Lepidopterologica and

we welcome you to the first issue of Nota published by Pensoft. In this issue you can read about

an unexpected discovery of Pyrgus malvoides in the Czech Republic, the revision of Naarda

(Erebidae), a detailed examination of the Alpine populations of the butterfly Erebia euryale,

an interesting new species of larentiine geometrids from the Kopet-Dagh Mountains, reduced

mouth parts in coleophorids, the taxonomic status of two species of Ochromolopis (Epermenii-

dae), the description of the previously unknown female of Ethmia cribravia (Elachistidae), a

remarkable new species of gelechiids from Turkmenistan, difficulties of identifying species of

Melitaea based on their wing morphology, and about choreutids of Madeira.

Nota Lepi. 37(1) 2014: 3-8 | DOI 10.3897/n1.37.7936

The first record of Pyrgus malvoides (Elwes & Edwards, 1897)

in the Czech Republic (Lepidoptera, Hesperiidae): an accidental

introduction?

VLADIMIR HULA', JANA NIEDOBOVA!”

1 Department of Zoology, Faculty of Agronomy, Mendel University, Zemedelska 1, CZ-613 00, Brno, Czech

Republic; Hula@mendelu.cz

2 Department of Forest Botany, Dendrology and Geobiocoenology, Faculty of Forestry and Wood Technology,

Zemedelska 3, CZ-613 00, Brno, Czech Republic

http://zoobank.org/A10070BE-B52B-4B2E-AD78-3777910CDF97

Received 7 August 2013; accepted 14 October 2013; published: 15 June 2014

Subject Editor: Zdenëk F. Fric

Abstract. The Mediterranean skipper, Pyrgus malvoides (Elwes & Edwards 1897), is newly recorded for

the Czech Republic, some 400 km away from its nearest known population. The specimen was collected

in June 1993 in Hejna, Bohemia and was discovered in the collection of Oldfich Jakes during a revision

of other Pyrgus species. The presence of this species elsewhere in the region could not be established and

the best current explanation for this record is that it was an accidental introduction.

Introduction

In recent decades, only a few new butterfly species have been recorded in the fauna of the

Czech Republic. For example, in the last ten years Pyrgus trebevicensis (Warren, 1926) (Benes

et al. 2001; Hané 2011) and Spialia orbifer (Hiibner, 1823) (Sumpich 2011) (both Hesperiidae)

were recorded for the first time in the Czech Republic, and just two additional species were dis-

covered during the last 20 years: Leptidea reali Reissinger, 1989 (Pieridae) and Hyponephele

lupinus (Costa, 1836) (Nymphalidae); the first one very abundant and the latter already extinct,

discovered as a museum specimen (Krälicek and Povolny 1992; Laëtüvka et al. 1995).

During a revision of skippers belonging to the genus Pyrgus (Hübner, 1819), an unusual

specimen was found in the collection of Oldiich Jakes with the collecting data: CZ — Bohemia,

Hejnä, 1.vi.1993, Jake’, O. Igt. (see Fig. 1). This locality is in the foothills of Sumava Mts. near

the town of SuSice (49°17’13”N, 13°40’18”E); it is covered by supramontane xeric vegetation

on limestone, and is part of the Pucanka Nature Reserve.

It was immediately noticed that this male specimen was unusually large and had different

colouration from the common Pyrgus malvae (Linnaeus, 1758), which is known to occur in the

region. The wingspan of this specimen is 16 mm, which is about three millimetres greater than

the 11-13 mm wingspan described by Slamka (2004) for P malvae, and it has a light brown

colour (Figs 2, 3), which is atypical for P malvae in the Czech Republic. After this discovery,

the first author decided to examine more material from across the Czech Republic. The results

+ HuLA & NIEDOBOWA: The first record of Pyrgus malvoides (Elwes & Edwards, 1897)...

of this investigation, presented below, led us to conclude that this unusual specimen is actually

P. malvoides (Elwes & Edwards 1897).

Methods

The examined collections are listed below. All material is from the Czech Republic. The first

author focused on the determination of different specimens of P. malvae from the region, study-

ing 13 specimens from 6 localities near the foothills of Sumava Mts, and then the first author

examined more material from other localities in the Czech Republic to attempt to find another

specimen of P malvoides. In addition to the examination of external characters, all genitalia

were dissected and determined by the first author (det. V. Hula).

Abbreviations

Collections

AP Alois Pavlicko, private collection, in Prachatice, Czech Republic

MM Miroslav Mikat, private collection, in Hradec Krälové, Czech Republic

MZMB Moravian museum Brno, in Brno, Czech Republic

OJ Oldïich Jake, private collection, in Brno, Czech Republic

PS Pavel Skala, private collection, in Praha, Czech Republic

RMM Regional museum Mikulov, in Mikulov, Czech Republic

VCMHK East-bohemian museum Hradec Krälové, in Hradec Kralové, Czech Republic

VCMP East-bohemian museum Pardubice, in Pardubice, Czech Republic

VH Vladimir Hula, private collection, in Brno, Czech Republic

ZCMP West-bohemian museum Pilsen, in Pilsen, Czech Republic

ZFF Zdenék Faltynek Fric, Institute of Entomology, in Ceské Budéjovice, Czech Republic

ZN Zdenék Navratil, private collection, in Brno, Czech Republic

Other abbreviations

NPR National nature reserve

PP Nature monument

PR Nature reserve

Material examined. Bohemia: Rakovnik — Papirna, 4.vi.1991 (1 ex), leg., det. et coll. VH. PR Niva u Zdirce nad

Doubravou, 28.v.2003 (2 ex), leg., det. et coll. WH. Brazec, 7.vi.2002 (1 ex), leg., det. et coll. VH. Kramolin - Podhora,

20.vi.2003 (1 ex), leg., det. et coll. VH. Plzen, 28.iv.1957 (2 ex), 9.v.1946 (1 ex.), 25.iv.1966 (1 ex), 27.1v.1966 (1 ex.),

leg. VI. Skala, coll. ZCMP; 23.iv.1960 (1 ex), leg. Fraj, coll. ZCMP; Dub. vrch, 9.v.1955 (1 ex), 14.1v.1946 (2 ex),

leg. VI. Skala, coll. ZCMP. Dobrany, Brez. vrch, 20.v.1956 (1 ex), leg. VI. Skala, coll. ZCMP; 28.v.1956 (1 ex), leg.

VI. Skala, coll. ZCMP. Chlistov, 6.vi.1954 (1 ex), leg. VI. Skala, coll. ZCMP; Jarov, 22.v.1925 (1 ex), coll. ZCMP;

Vladaï, 1908 (1 ex), coll. ZCMP; NPR VySenské kopce, 20.vi.2001 (1 ex), leg. et coll. ZFF; Hfedle, 27.v.1995 (1 ex),

leg. et coll. ZFF; Hlinky, 19.vi.1994 (1 ex), leg. J. Franz, coll. WH; Police nad Metuji, 9.iv.1944 (1 ex), leg. G. Petr,

coll. VCMHK; Srbsko, 21.v.1961 (1 ex), 9.v.1962 (1 ex) coll. VCMHK; Cihana, 22.v.1971 (1 ex), 9.vi.1972 (1 ex),

leg. J. Franz, coll. VH; Nové Kounice, 30.v.1992 (1 ex), leg. J. Franz, coll. VH; Rybniéna, 28.1v.1996 (1 ex), 29.v.1991

(1 ex), 31.v.1991 (1 ex), leg. J. Franz, coll. VH; Javorna, 3.vi.1972 (1 ex), leg. J. Franz, coll. VH; Pila, 27.vi.1991 (1

ex), leg. et coll. VH; Horni TaSovice, 31.v.1972 (1 ex), leg. Franz J., coll. VH; Tepliëka, 14.v.1992 (1 ex), leg. Franz J.,

coll. VH; Némecky Chloumek, 9.vi.1972 (1 ex), leg. Franz J., coll. VH; Ceska Trebovä, 20.v.1964 (1 ex), leg. Siroky

L., coll. VCMHK; PP Ban, Hradéany, 1.v.2001 (1 ex), leg. et coll. MM; PP Na Plachté, Hradec Krälové, 15.v.2001 (1

ex), 25.v.2002 (1 ex), leg. et coll. MM; Vysoka and Labem, 8.v.1976 (1 ex), leg. et coll. MM; Slezia, TéSin, 18.v.1930

(1 ex), coll. VCMHK; Oblik, 2.v.1987 (1 ex), leg. Pacl, coll. VCMHK,; Prachatice, 18.v.1982 (1 ex), 27.v.1978 (1 ex),

20.vi.1978 (1 ex), 26.iv.1981 (1 ex), 29.11.1980 (1 ex), 10.vii.1977 (1 ex), leg. et coll. AP; Setun, Prachatice, 24.v.1985

Nota Lepi. 37(1) 2014: 3-8 5

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Figures 1-5. 1. Grid square map of the Czech Republic with marked square of record of Pyrgus malvo-

ides (Elwes & Edwards, 1897). 2-3. The collected specimen Pyrgus malvoides, CZ — Bohemia, Hejnä,

1.vi.1993, Jakes, O. let. et coll., Hula, V. det. 4. Male genitalia (valva, lateral view) of collected specimen

P. malvoides. 5. General view from the collecting locality of limestone quarry.

(1 ex), leg. et coll. AP; Irü Dvür, Prachatice, 9.v.1986 (1 ex), leg. et coll. AP; Lipa, Prachatice, 10.vii.1977 (1 ex), leg.

et coll. AP; Kunéticka hora - Raby, 9.v.1994 (1 ex), leg. J. Macourek, coll. VCMP; Stfizovicky vrch, Usti nad Labem,

8.v.2002 (2 ex), leg. et coll. VH; PR Puéanka, Hejnä, 1.iv.1993 (1 ex), leg. OJ, coll. VH; Kliëava, (1 ex), leg. Dr. VI.

Vala, coll. RMM. Moravia: Filipovské üdoli, 5.vi.1951 (1 ex), leg. Dr. VI. Vala, coll. RMM; Lideïovice, 11.v.1950

6 HULA & NIEDOBOWA: The first record of Pyrgus malvoides (Elwes & Edwards, 1897)...

(2 ex), leg. Dr. VI. Vala, coll. RMM; Luhacovice, 20.v.1946 (1 ex), 13.v.1951 (lex), leg. Dr. VI. Vala, coll. RMM;

Uherské Hradisté, 9.v.1949 (1 ex), leg. Dr. VI. Vala, coll. RMM; Kfriscanovice, 13.v.1947 (1 ex), leg. Dr. VI. Vala, coll.

RMM; Bfeclav, 13.vi1.1979 (1 ex), leg. E. Sandtner, coll. RMM; Valtice, 28.1v.1968 (1 ex), 10.v.1975 (1 ex), leg. E.

Sandtner, coll. RMM; Milovice, 18.v11.1962 (1 ex), leg. Minarik, coll. RMM; NPR Malhotky, 25.vi.2004 (1 ex), leg.,

det. et coll. VH; Moravsky Krumlov, 3.v1.1944 (1 ex), coll. MZMB; Senorady, 29.v.1935 (1 ex), leg. Lemberk, coll.

MZMB; Habrüvka, 26.v.1982 (1 ex), leg. Kotlan and Veprek, coll. MZMB; Hädy, 1962-1982 (1 ex), leg. Kotlan and

Veprek, coll. MZMB; Macoëskä Stran, 4.v1.1994 (1 ex), leg. et coll. PS; Brno - Medlanky, 30.iv.1994 (1 ex), leg. J.

Macourek, coll. VCMP; 15.v.1983 (1 ex), leg. R. Cech, coll. ZN. Macoëskä Strän, 20.vi.1995 (1 ex), leg. J. Macourek,

coll. VCMP; Zeleëice, 30.v.1991 (1 ex), leg. J. Macourek, coll. VCMP; Perna, CHKO Palava, 9.vi.2006 (1 ex), leg. et

coll. OJ; Mohelno, 29.v.1977 (1 ex), leg. et coll. ZN; Kamenny vrch, Brno, 6.v.1979 (3 ex), leg. et coll. ZN; SobéSice,

11.v.1977 (1 ex), leg. et coll. ZN; Radéjov, 25.1v.1948 (4 ex), 26.vi.1954 (1 ex), leg. Dr. VI. Vala, coll. RMM.

Results and discussion

Twelve collections from 58 sites in the Czech Republic were studied. This included 88 speci-

mens of P. malvae from a wide range of territories in the Czech Republic, and even one spec-

imen of P. malvae from the same locality as the P malvoides was found. A thorough search

uncovered no other specimens of P. malvoides.

Pyrgus malvoides is a typical west-Mediterranean species with its distribution ending

in Slovenia, Italy, westernmost Austria, Liechtenstein, Switzerland and France (Kaufmann

1955; Aistleiner 1996; Lepidopterologen Arbeitsgruppe 1997; Karsholt & Nieukerken 2011).

Some authors consider the taxon P. malvoides only as a subspecies of P. malvae (see Tolman

& Lewington 1997; Kudrna et al. 2011). These taxa are able to hybridize on the border of

their distribution (Guillaumin 1962, 1971) and the hybrids show characters of both species

(Albeti 1956). De Jong (1972, 1987) studied the whole complex of species in the P malvae

group and summarised the distribution of all species, specific characters and morphological

phylogeny of the group. In this work clear morphological differences in male and female

genitalia can be found. The specimen of P malvoides that we discovered in the collection of

Oldfich Jakes has no marks of hybridisation (Fig. 4).

The origin of the specimen is unclear. All available material of P malvae from the region

(the foothills of Sumava Mts., the mainly limestone regions and some other places across the

whole Czech Republic) was investigated. It is very unlikely that undiscovered populations exist

because the butterfly fauna of the Czech Republic has been very well investigated (Benes and

Konvicka 2002). It is also unlikely that P malvoides had flown in from the nearest population,

which is about 400 kilometres distant.

It is likely that this specimen of P malvoides was accidentally introduced into the locality.

There are several possibilities, but the most probable seems to be the introduction by mining

techniques, as the locality is in the vicinity of a very large limestone quarry (Fig. 5) owned by

the French company Lafarge. Nevertheless, it is an interesting record of this species in Central

Europe, and illustrates the importance of paying close attention even to species as common as

Pyrgus malvae.

Nota Lepi. 37(1) 2014: 3-8 7

Acknowledgements

We are grateful to Oldfich Jakeë for providing the material of P malvae and P. malvoides and all infor-

mation about collecting. We are grateful to all who shared their Pyrgus collections. Thanks to anonymous

referees for their comments. Many thanks to Marina Isaac for proofreading. This study was funded by

the Internal Grant Agency of Agronomical Faculty (IGA TP6/2013 and TP 7/2014) and the Faculty of

Forestry and Wood Technology (73/2013) of Mendel University.

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Sumpich J (2011) Motyli Närodnich parkü Podyji a Thayatal (Die Schmetterlinge der Nationalparke Podyji

und Thayatal). Ist ed. National Park Podyji Administration Office (Znojmo), Znojmo. 428 pp.

Tolman T, Lewington R (1997) Collins field guide of butterflies of Britain & Europe. Ist ed. Harper Co-

llins Publishers, London. 320 pp., 104 pls.

Nota Lepi. 37(1) 2014: 9-18 | DOI 10.3897/n1.37.7957

Revision of the Palaearctic and Oriental species of the genus Naarda

Walker, 1866 (Lepidoptera, Erebidae, Hypeninae). Part 3.

Description of three new species from Asia

BALAZS TOTH', LASZLO RONKAY!

1 Collection Lepidoptera, Hungarian Natural History Museum, Baross utca 13, HU-1083 Budapest, Hungary;

balazs0toth@gmail.com, ronkay@nhmus.hu

http://zoobank.org/B2D8E504-EBDS&-4A C6-B2A9-A75B1320B6D2

Received 30 April 2013; accepted 21 October 2013; published: 15 June 2014

Subject Editor: Alberto Zilli

Abstract. We revise the Naarda egrettoides species-group and describe and illustrate three new Naarda

species: N. ardeola sp. n. (Thailand), N. egrettoides sp. n. (Thailand) and N. pocstamasi sp. n. (Vietnam).

Introduction

The authors began revising the genus Naarda in 2000 with the study of the Noctuidae s.l.

fauna of Taiwan. In the process, it was discovered that more species occur in Taiwan than was

formerly indicated in the literature (Wileman 1915; Strand 1920; Heppner & Inoue 1992). Sub-

sequent studies carried out on the extensive Naarda material from different parts of eastern and

south-eastern Asia led to an unexpected increase of the known species richness of the genus,

with the number of Asiatic species exceeding 80.

These results were first presented by BT (2010) at the IX" European Congress of Entomolo-

gy (Lepidoptera taxonomy and biogeography section), announcing the publication of the unde-

scribed species in the near future before the revision of the whole genus, including its African,

Eurasiatic and Australian species, could be accomplished.

The first two parts of the relevant series of articles deal with the taxonomy and biogeography

of the genus Naarda Walker, 1866 and contain an overview of this large and diverse group,

including the general morphological characterisation of the main lineages and the descriptions

of altogether 39 new species from eastern and south-eastern Asia (Toth & Ronkay in press a, b).

The present paper contains the survey of a compact and in the male genitalia highly apo-

morphic species-group, the Naarda egrettoides lineage, including the description of three new

species from the south-eastern border zone of the Palaearctic region.

10 TOTH & RONKay: Revision of the Palaearctic and Oriental species of the genus Naarda Walker...

Material and methods

All but one specimen of the material examined belonging to the Naarda egrettoides lineage

are deposited at the Hungarian Natural History Museum, Budapest (HNHM); a specimen of a

currently unnamed species from eastern China is held by the Alexander Koenig Museum, Bonn

(ZFMK).

Genitalia dissections were made using the standard method of preparation resulting in per-

manent microscopic slides. Genital structures were macerated in 10% KOH solution and lactic

acid, stained with alcoholic solution of Eosine and mounted in Euparal. Eosine was preferred

in contrast to Chlorazol Black because of the dominance of chitinous structures in the genitalia.

The genital terminology follows the detailed explanation given in the first part of the series of

papers (Toth & Ronkay in press a).

Abbreviations

BMNH The Natural History Museum, London

HNHM Hungarian Natural History Museum, Budapest

MNHN Museum National d’Histoire Naturelle, Paris

RL genitalia slide of Läszlö Ronkay

TB genitalia slide of Balazs Toth

ZFMK Alexander Koenig Museum, Bonn — Zoologisches Forschungsinstitut und Museum Alexander Koenig

Characterisation of the Naarda egrettoides species-group

The species-group is very compact and comprises three closely related species, which are eas-

ily separable from members of all other lineages of the genus by the features of the males (see

below); the females are also characteristic but display no key features except for the colouration

and the fine and less prominent dark markings.

External morphology. The diagnostic features are the long bipectinate male antenna, the

very long, porrect labial palpi of both sexes, the rather large size, the long and relatively broad

forewings with straight costa, and the weak, fine darker markings on a pale greyish or brown-

ish ground colour. The rami of the antenna are at the maximum of its width 9-10 times longer

than the axis of the antenna; the apical segments lack the rami. The palpi are ca. 5 times as

long as the diameter of the eye, with the third segment comparatively longer, and more elon-

gated than in most other members of the genus. The wingspan is 19-22 mm.

Male genitalia. The most prominent autapomorphy of the group is the structure of the uncus

(see Figs 1, 3, 5), which appears as a head of a long-billed bird. The uncus has a large, rounded

subbasal dorsal bulb bearing a small but acute frontal spine and a large bundle of long hairs

standing apart (like a forelock) and long, straight, apically finely dilated and rounded distal sec-

tion. Other group features of the clasping apparatus are the large, broad tegumen, the well-de-

veloped and strong transtilla and the variably broad, generally triangular valvae with which

the entire structure resembles a flying bird (wader). Phallus short, thick and straight, vesica

basally broad and inflated, ductus ejaculatorius directed forward in the axis of the phallus and

vesica. Cornutus variably large, flattened and somewhat ‘sponge cake’-like, situated subbasally

at ventral side.

Nota Lepi. 37(1) 2014: 9-18 1]

Synopsis

ardeola sp. n. (Northern Thailand)

pocstamasi sp. n. (Northern Vietnam)

egrettoides sp. n. (Northern Thailand)

Descriptions of the new species

Naarda ardeola sp. n.

http://zoobank.org/AB56DE34-4DA4-4BE4-87A D-DCD915EC5040

Figs 1, 2, 7, 8

Holotype. À, ‘Thailand: Prov. Chiang Mai | between Chiang Dao and Kariang, 900 m | 99°48’E, 19°25’N, 26.x.2002 |

leg. B. Herczig & G. Ronkay’ slide No. RL7898m (coll. HNHM).

Paratypes. Thailand: 29, data as holotype, slide No. TB398f; 19, ‘Prov. Chiang Mai | 1600m, between Fang and

Nor Lae | 99°09’E, 20°02’N, 28.x.2002 | leg. B. Herczig & G. Ronkay’ (coll. HNHM); 19, ‘50 km NW Mae Hong-

Son | by Shan, 800 m, 14.vii.2003 | leg. M. Fibiger’ slide No. TB424f (coll. HNHM).

Description. Wingspan 20-21 mm, length of forewing 10-11 mm. Antennae bipectinate

in male but apical segments lack rami, filiform and ciliate in female; longest male rami at mid

third, 9 times longer than diameter of flagellum, rami with cilia as long as diameter of flagel-

lum; in female each segment with two cilia as long as diameter of flagellum. Labial palps simi-

lar in both sexes; their length 5 times diameter of eyes; 3rd segment relatively long and narrow,

its tip pointed, light; 2nd segment broad, dorsal scales long, their length descending towards

tip. Scale-hood of vertex broad, tapering, in male relatively long, apically slightly bifurcate, in

female long, its tip rounded. Base of male forewing with scent-organ built up of long, hairy

scales. Characteristic wing pattern features: forewing costa minutely concave in male; pattern

similar in both sexes: ground colour light brown, subterminal line slightly paler, slightly sinu-

ous, with some blackish dots at inner side; postmedial line slightly darker than ground colour,

indistinct; reniform stigma big, oval, deep ochreous, hardly visible, with small blackish dot at

bottom section; orbicular stigma also deep ochreous, indistinct. Hindwing slightly paler than

forewing, with slightly more conspicuous fasciae.

Male genitalia (Fig. 1). Uncus relatively long, straight, tapering, its tip rounded, basal part

containing globular bulb with short spine close to elongate part of uncus, and a hairy area op-

positely. Scaphium very short, straight. Tegumen longer than vinculum. Transtilla enormously

wide; its basal part is the broadest in the genus. Saccus not visible. Base of valva narrow; valva

slightly tapering, its tip very narrow, rounded. Sacculus wide, short, distally fused with the

homogeneous fused structure constructing the apical two-thirds of valva. Phallus elongate,

straight, not tapering; carina with two narrow, curved processes; vesica oval, smooth, with

long, broad diverticulum and strong, straight cornutus with rounded tip.

Female genitalia (Fig. 2). Ovipositor lobes slightly elongate, angular. Apophyses quite broad

and long, apophyses posteriores 1.3 times longer than apophyses anteriores. Lamella antevagi-

nalis short, angular. Sinus very short and wide. Sternum A7 with two strongly sclerotised nar-

row bands laterally from ostium, and a proximally located sclerotised triangular field. Ductus

12 TOTH & RonKAY: Revision of the Palaearctic and Oriental species of the genus Naarda Walker...

Figures 1-6. Male and female genitalia ofthe species ofthe Naarda egrettoides species-group. 1. Naarda

ardeola sp. n. (RL7898m); 2. Naarda ardeola sp. n. (TB398f); 3. Naarda pocstamasi sp. n. (RL10762m);

4. Naarda sp. (TB443f); 5. Naarda egrettoides sp. n. (RL7899m); 6. Naarda egrettoides sp. n. (TB390f).

Nota Lepi. 37(1) 2014: 9-18 13

bursae short, membranous, colliculum small. Corpus bursae elongate, not tapering, with dense

scobination except for a dorsal area.

Diagnosis. The external appearance of N. ardeola is nearly unique: it can only be con-

fused with N. pocstamasi sp. n., although the ground colour of N. ardeola is somewhat more

yellowish, and the transverse lines are less conspicuous than in the latter taxon. The male

genitalia of N. ardeola differ from those of N. pocstamasi by the more angular basal bulge

and the slightly broader elongate section of the uncus, the longer and considerably narrower

valva, and the more simple structure of the vesica of phallus, with shorter and narrower

cornutus. The very elongate valvae in the male genitalia are somewhat similar to those of

N. serra Holloway, 2008, but the other characters are very different: the basal part of uncus

is much broader, with a bulb which is absent in N. serra, the scaphium is much shorter, and

the base of transtilla is much broader. The penicular processes, being typical of N. serra,

are absent in N. ardeola. The apical half of the valva is made of different structures: in N.

ardeola it is the complete fusion of costa, cucullus and harpe, while in N. serra this part is

the free cucullus.

Etymology. The heron-shaped male genitalia inspired the specific name.

Distribution. The species is known from the mountains of northern Thailand.

Naarda pocstamasi sp. n.

http://zoobank. org/3364C604-943 B-46B5-B4C7-4F604BA28DF2

Figs 3, 9

Holotype. À, ‘Vietnam, Prov. Lao Cai | Sa-pa, 1650 m | 23.ix.1963 | T. Pöcs’ ‘lieu herb. second. | à la lumière” slide

No. RL10762m (coll. HNHM).

Description. Wingspan 22 mm, length of forewing 11 mm. Antennae bipectinate (apical

segments also), longest male rami at mid third, maximum 10 times longer than diameter of fla-

gellum, rami with cilia ca 1.5 as long as diameter of flagellum. Labial palps slightly longer than

5 times diameter of eyes; 3rd segment quite long and narrow, its distal half lighter; 2nd segment

relatively narrow, slightly tapering. Scale-hood of vertex broad and long, triangular and apical-

ly pointed. Fore- and midtibiae densely hairy. Characteristic wing pattern features: forewing

costa not concave; ground colour light brownish grey (the specimen is somewhat worn), termi-

nal line prominent, fragmented to blackish dots, subterminal, postmedial and antemedial lines

present but indistinct, slightly sinuous; reniform stigma inconspicuous, medium-sized, broad,

somewhat 8-shaped, with traces of dark edge, brownish ochreous with blackish dot at bottom

third and traces of a brownish line in the axis; orbicular stigma rounded with indistinct blackish

edge, its colouration like that of reniform, with some dark scales in its centre. Colouration and

pattern of hindwing like that of forewing.

Male genitalia (Fig. 3). Uncus medium long, straight, its basal part a densely haired, elongate,

apical part evenly narrow, its tip rounded. Tegumen as long as vinculum. Saccus broad-based,

long, gradually tapering, its tip rounded. Juxta large, angular. Transtilla large, broad. Saccus

not visible. Valva long, medium wide, gradually tapering, but dorsal edge slightly sinuous. Val-

val tip slightly rounded-truncate. Sacculus with very broad but short, densely haired basal half

14 TOTH & RoNKAY: Revision of the Palaearctic and Oriental species of the genus Naarda Walker...

Figures 7-12. Adults of the species of the Naarda egrettoides species-group. 7. Naarda ardeola sp. n.

male; 8. Naarda ardeola sp. n. female; 9. Naarda pocstamasi sp. n. male; 10. Naarda sp. female; 11. Naar-

da egrettoides sp. n. male; 12. Naarda egrettoides sp. n. female.

and narrow, straight distal half connected to apical fused structure. Phallus very thick, evenly

wide, straight; vesica globular, longitudinally striated with a large, straight, rounded, somewhat

tongue-like cornutus.

Diagnosis. The external appearance of N. pocstamasi is similar to that of N. egrettoides

sp. n., but the ground colour is lighter, and the transverse lines are more prominent in N. poc-

stamasi, making it easily recognisable. The male genitalia of this new species are very similar

Nota Lepi. 37(1) 2014: 9-18 15

to those of N. egrettoides, but the basal bulb of uncus is more elongate, the valva is somewhat

narrower and more elongate, the costa is more sinuous, the phallus is shorter and thicker, and

the cornutus is larger. In comparison with those of N. kinabaluensis Holloway, 2008, the uncus

is much narrower and shorter, the scaphium is much shorter, the valva is somewhat narrower

and its tip is pointed instead of being truncate.

Etymology. This new species is dedicated to its collector, a famous Hungarian botanist,

Tamas Pocs.

Distribution. Northern Vietnam.

Remarks. This species appears to be the closest hitherto known relative of N. egrettoides

sp. n. according to the male genitalia.

Naarda sp. near egrettoides

Figs 4, 10

Material examined. ©, ‘China: Shanghai | Prov. Kiangsu | leg. Hone’ slide No. TB443f (coll. ZFMK).

Morphology. Wingspan 21 mm, length of forewing 10 mm. Antennae lost except for a few

basal segments, those being ciliate, without rami or crests, length of cilia half times diameter of

flagellum. Labial palps longest in the genus: length slightly less than 6 times diameter of eyes;

3rd segment relatively long, narrow, its apical third light; 2nd segment broad, dorsal scales

very short (or maybe worn). Scale-hood of vertex broad-based, relatively short, its tip pointed.

Characteristic wing pattern features: costa minutely concave; the only one known specimen

is worn and possibly faded, ground colour greyish brown; transverse lines indistinct; bottom

half of postmedial line hardly visible, dark, narrow, probably strongly angled inwards below

cell; reniform stigma hardly visible, quite small, oval, dark ochreous, with a big blackish dot at

the bottom third; orbicular stigma also hardly visible, small, dark ochreous. Ground-colour of

hindwing like that of forewing, with two fasciae.

Female genitalia (Fig. 4). Ovipositor lobes small, angular. Apophyses long; apophyses pos-

teriores ca 1.5 times longer than apophyses anteriores. Lamella antevaginalis angular, scle-

rotised, sinus absent. Ductus bursae broad but short, membranous, its mouth to corpus bursae

located at the dorsal side of corpus bursae. Corpus bursae elongate, pyriform, posterior 2/3 part

being scobinate, becoming sparser anteriorly, posterior 1/3 part with a very broad but quite

short, smooth appendix.

Diagnosis. The wing shape of N. sp. nr. egrettoides is similar to that of N. egrettoides and N.

ardeola, but the reniform stigma seems to be narrower, and the orbicular is smaller than in the

latter taxa. The female genitalia are somewhat similar to those of N. barlowi Holloway, 2008,

especially in the configuration of the appendix bursae, which is broad and situated close to the

posterior tip of corpus bursae. This species lacks the sinus and the colliculum of ductus bursae

and the corpus bursae is only sparsely scobinate, in contrast to N. barlowi.

Distribution. South-eastern China (Prov. Jiangsu), at the transitional zone of the Palaearctic

and Oriental Regions.

Remarks. The long labial palps and the main structure of the female genitalia are similar to

those of N. egrettoides and N. ardeola, but more material, and especially the knowledge of the

16 TOTH & RONKay: Revision of the Palaearctic and Oriental species of the genus Naarda Walker...

male genitalia, are essential to decide on the more exact taxonomic assignment of this individ-

ual. To make the situation even more difficult, the type locality has apparently been destroyed

by development in the last seventy years.

Naarda egrettoides sp. n.

http://zoobank.org/B7E4A 1BB-DAA1-4CA0-A952-2COAEFCEFB78

Figs 5, 6, 11, 12

Holotype. À, ‘Thailand: Prov. Chiang Mai | 4 km S Kop Dong, 1800 m | 99°03’E, 19°52’N, 6.xi.2002 | leg. B. Herczig

& G. Ronkay’ slide No. RL7897m (coll. HNHM).

Paratypes. Thailand: 14, 39, ‘Prov. Nan | Doi Phu Kha NP | between Pua and Bo Luang, 1350 m | 101°05’E,

19°12’N, 3.xi.2002 | leg. B. Herczig & G. Ronkay’ slide No. RL7899m (coll. HNHM); 24, 29, ‘Prov. Chiang Mai |

1600m, between Fang and Nor Lae | 99°09’E, 20°02’N, 28.x.2002 | leg. B. Herczig & G. Ronkay’ slide No. TB390f

(coll. HNHM).

Description. Wingspan 19-22 mm, length of forewing 10—11 mm. Antennae bipectinate in

male but apical segments lacking rami, filiform and ciliate in female; longest male rami at mid

third, 9 times longer than diameter of flagellum, rami with cilia as long as diameter of flagel-

lum; in female each segment with four cilia on each side shorter than diameter of flagellum.

Labial palps similar in both sexes; length 5 times diameter of eyes; 3rd segment quite long and

narrow, its tip pointed, light; 2nd segment broad, dorsal scales long, in male length of scales

descending towards tip, in female scale length uniform. Scale-hood of vertex broad-based and

long in both sexes, in male its tip finely bifurcate, in female rounded. Characteristic wing pat-

tern features: forewing costa not concave in male; sexes similar; ground colour greyish brown,

subterminal and postmedial lines parallel, sinuous, inner part of them dark grey, outer part

mouse-grey; medial and antemedial lines also present but indistinct; reniform stigma large,

thick, oval, deep ochreous (honey-coloured) with blackish dot at bottom third; orbicular stigma

longitudinally elongate, also honey-coloured, with indistinct blackish edge. Hindwing slightly

paler than forewing, postmedial line more visible than subterminal line.

Male genitalia (Fig. 5). Uncus relatively long, straight, apically slightly dilated, its tip round-

ed, basal bulb oval, spine on bulb tiny, situated in front of the connection of bulb and tegumen.

Scaphium very short, straight. Transtilla very broad, its base wide. Tegumen slightly longer

than vinculum. Saccus not visible. Valva relatively broad-based, triangular, gradually tapering,

its tip pointed. Dorsal edge of valva slightly concave. The fused structure present at distal half

of valva. Sacculus with broad but short basal half and narrow, straight distal half connected to

apical fused structure. Phallus thick, slightly curved, slightly tapering towards carina. Vesica

globular, longitudinally striated, with strong, straight, broad and apically rounded, somewhat

tongue-shaped cornutus (seen from edge on the figure).

Female genitalia (Fig. 6). Ovipositor lobes slightly elongate, relatively small, angular. Ap-

ophyses narrow, apophyses posteriores very slightly (1.1 times) longer than apophyses ante-

riores. Lamella antevaginalis short, angular. Sinus absent. Sternum A8 with small triangular

plate arranged anteriorly. Ductus bursae broad, short, membranous. Corpus bursae composed

from of two equal-sized globular halves connected by narrow region; posterior part densely

Nota Lepi. 37(1) 2014: 9-18 17

scobinate by tiny spines, anterior part smooth except for small, sparsely scobinate area near the

connection to distal part.

Diagnosis. Based on their morphology, it appears that the sister-species of N. egrettoides

is N. pocstamasi; their detailed comparison is given under the diagnosis of the latter. N. egret-

toides differs externally from the other close relative, N. ardeola, by the darker, more greyish

colouration. In the male genitalia, N. egrettoides and N. pocstamasi have, in comparison with

N. ardeola, much broader valvae, more rounded basal bulge of uncus and much larger cor-

nutus in the vesica. The male genitalia of N. egrettoides are somewhat similar to those of N.

ineffectalis (Walker, 1858), but in the new species the tip of uncus is not concave while the

base of uncus is much broader, the juxta is smaller, the valva is more elongate, the sacculus is

broader, the cornutus of phallus is straighter and the vesica is larger and more globular than in

N. ineffectalis.

Etymology. Egrettoides = egret-like; the shape of male genitalia is similar to a flying egret.

Distribution. The mountains of northern Thailand.

Remarks. The longitudinally elongate shape of the orbicular stigma is a rare feature in the

genus.

Acknowledgements

We are grateful to Dieter Stiining (ZFMK), Martin Honey (BMNH) and Joél Minet and Jerome Barbut

(MNHN) for access to the material of their institutes and the loan of several Naarda specimens, which

were essential to our work. The authors express their thanks to Béla Herczig, Gabor Ronkay, and the late

Michael Fibiger for lending Naarda specimens for examination. The first author would like to express

his thanks to the leaders of the Hungarian Natural History Museum for obtaining research authorisation

for the examination of the Naarda material hosted at HNHM. He is indebted also to the staff members

of the Lepidoptera Collection for their generous help during his research. The visits of Laszlo Ronkay

to the BMNH and the MNHN were supported by the SyntheSys Project (Grant Nos GB-TAF-2656 and

FR-TAF-562).

References

Heppner JB, Inoue H (Ed) (1992) The Lepidoptera of Taiwan, vol. 1, part 2: Checklist: 1-1, 1-276. Asso-

ciation for Tropical Lepidoptera & Scientific Publishers, Gainsville, Florida, USA.

Holloway JD (2008) The moths of Borneo: family Noctuidae, subfamilies Rivulinae, Phytometrinae,

Herminiinae, Hypeninae, Hypenodinae. Malayan Nature Journal 60 (1-4): 1-268.

Poole RW (1989) Noctuidae. In Heppner JB (Ed). Lepidopterorum Catalogus (New Series) 118. E. J. Brill

/ Flora & Fauna Publications, Leiden, New York, Kobenhavn & Köln, pp. 501-1013.

Strand E (1920) H. Sauter’s Formosa-Ausbeite: Noctuidae II., nebst Nachtrage zu den Familien Arctiidae,

Lymantriidae, Notodontidae, Geometridae, Thyrididae, Pyralidae, Tortricidae, Gelechiidae und Oeco-

phoridae. Archiv für Naturgeschichte 84 A 12: 102-197.

Töth B (2010) The Asian species of the genus Naarda Walker, 1866: review and taxonomy (Lepidop-

tera: Noctuidae, Hypeninae). Oral presentation. IX" European Congress of Entomology, Budapest,

Hungary.

Toth B, Ronkay L (in press a) Revision of the Palaearctic and Oriental species of the genus Naarda

Walker, 1866 (Lepidoptera: Erebidae, Hypeninae). Part 1. Taxonomic notes and description of 29 new

species from Asia. Oriental Insects.

18 TOTH & RONKay: Revision of the Palaearctic and Oriental species of the genus Naarda Walker...

Toth B, Ronkay L (in press b) Revision of the Palaearctic and Oriental species of the genus Naarda

Walker, 1866 (Lepidoptera: Erebidae, Hypeninae). Part 2. Description of 10 new species from

Asia. Acta Zoologica Academiae Scientiarum Hungaricae.

Wileman AE (1915) New species of Noctuidae from Formosa. The Entomologist 48: 191-196.

Nota Lepi. 37(1) 2014: 19-36 | DOI 10.3897/n1.37.7935

Reproductive isolation and intraspecific structure in Alpine

populations of Erebia euryale (Esper, 1805) (Lepidoptera,

Nymphalidae, Satyrinae)

FRANS CUPEDO!

1 Processieweg 2, 6243 BB Geulle, Netherlands; frans@cupedo.eu

http://zoobank. org/C7D22F 60-7585-4970-BC35-CCES3FC2698C

Received 2 May 2013; accepted 31 January 2014; published: 15 June 2014

Subject Editor: Thomas Schmitt

Abstract. The subspecies of Erebia euryale (Esper, 1805) have been split into three groups based on

morphology, differing in male genital characters. Two of them, the euryale group and the adyte group,

are known to be strongly, but not completely, reproductively isolated. There is genetic evidence that

their separation preceded the differentiation of subspecies within the euryale group. No such data

exist on the third group, the recently recognized kunzi group. In this study, the degree of reproductive

isolation between the kunzi group and the other two groups is assessed. In three secondary contact

zones, a series of E. euryale populations were sampled in a transect perpendicular to the dividing line.

Morphological characteristics showed a clinal gradient along each transect. The steepest gradient was

found between the euryale and kunzi groups. Morphologically detectable introgression did not exceed

two kilometres. This is comparable to the situation described earlier in contact zones of the euryale and

adyte groups. In the contact area of the kunzi and adyte groups, the character gradient slope is more

gradual and the morphologically detectable introgression zone is at least five times wider. In contrast

to this, contact between subspecies belonging to the same group leads to virtually unrestricted morpho-

logical intermingling. It is concluded that the euryale group is reproductively more strongly isolated

from the other two groups than the kunzi group is from the adyte group, and that subspecies belonging

to the same group are interfertile to a high degree. It is argued that loss of genetic compatibility by long

term separation is the main cause of the reproductive isolation between groups, and that, consequently,

the actual intraspecific structure of E. euryale results from at least two, probably three, temporally

separated differentiation events.

Introduction

Erebia euryale (Esper, 1805) is a butterfly species with a highly disjunctive distribution and

considerable geographic variation. Both its genetic diversity and its distribution pattern have

been mainly shaped by climatic fluctuations during the Pleistocene (Schmitt and Haubrich

2008), as was the case in most Palaearctic organisms (Hewitt 1996; Comes and Kadereit

1998; Hewitt 1999, 2000; Kropf et al. 2002; Tribsch and Schönswetter 2003; Schönswetter

et al. 2005; Schmitt 2007, 2009). Climate induced range shifts, either latitudinal or altitudi-

nal, repeatedly led to area fragmentation and to retraction or expulsion into glacial refugia.

Long-term isolation in these refugia resulted in genetic divergence and, as a consequence,

in morphologic differentiation and decreased reproductive compatibility. Each of these three

aspects contributed to the actual knowledge of the intraspecific structure of E. euryale. Al-

20 Cupebo: Reproductive isolation and intraspecific structure in Alpine populations of Erebia euryale...

lozyme data suggest a two-level intraspecific structure. Schmitt and Haubrich (2008) deter-

mined the genetic distances between eleven E. euryale populations. These clustered into four

groups. The genetic distances between three of them were roughly equal, while the fourth

one was more distant. The authors suggested that this group had split off earlier, and that

the observed intraspecific genetic structure has resulted from two subsequent differentiation

events. This nested structure was less obvious in a study of mitochondrial DNA (Vila et al.

2011). The morphological structure of E. euryale shows two differentiation levels as well.

Cupedo (2010) analysed 72 populations, covering all described subspecies, and found them

clustering into three groups. These groups differ in male genital anatomy. They are known as

the euryale, adyte and kunzi group. The morphological and genetic structuring are concor-

dant: the genetically most distant cluster belongs to the adyte group; the three more coherent

clusters consistently represent different subspecies of the euryale group. The scarce existing

data on reproductive isolation fit into this pattern. Representatives of the euryale group and

the adyte group tend to remain separated in secondary contact zones (Rezbanyai-Reser 1991;

Sonderegger 2005). Transitional zones are narrow, and hardly contain any hybrids. Some

subspecies of the euryale group, on the other hand, build transitional zones of up to 40 km in

width (Cupedo 2010), mainly consisting of morphological hybrids.

Altogether, genetics, morphology, as well as the degree of reproductive isolation, support

the hypothesis of a two-level nested structure of E. euryale, at least as far as the euryale and

adyte groups are concerned. Little is known, though, on the kunzi group. Morphologically, it

has to be ranked in the first level of hierarchical differentiation because it differs considera-

bly from both the euryale and adyte groups in male genital anatomy. At present, genetic data

are lacking, and little is known about contact sites with the other two morphological groups

(Cupedo 2010).

The aim of the present study is to assess the degree of reproductive isolation between the

kunzi group and the other two groups, and to determine whether this is concordant with the

morphological traits. For this purpose, three known contact zones, one with the adyte group and

two with the euryale group, were intensively sampled. For each of these contact zones, three

questions were addressed: (i) Are hybrid populations present in the contact zone? If so, (11) what

is their composition, and (iii) does their composition show a clinal character gradient across the

contact zones? If the latter is found to be the case, the steepness of the cline will provide infor-

mation regarding the strength of reproductive barriers between the groups. Finally, all available

data on reproductive isolation in E. euryale are combined in order to establish whether or not

they support the hypothesis that two intraspecific differentiation levels exist.

Material and methods

The kunzi group

The kunzi group occupies a restricted but well-defined part of the Italian Alps (Fig. 1). Its dis-

tribution area comprises (i) the entire pre-Alps between Lake Como and the Valcellina, (11) the

Bergamasque Alps, (iii) the Southern Rhaetian Alps (Ortler, Adamallo-Presanella, Brenta and

Nonsberg Alps), and (iv) the Dolomites south of the Latemar-Focobon chain and west of the

Nota Lepi. 37(1) 2014: 19-36 21

Cordevole river. This territory is almost entirely bordered by insurmountable river valleys and

mountain chains.

Contact sites

Secondary contact with populations of other groups requires natural interruptions in this chain

of barriers. Four such “exchange windows” exist, three of which were investigated in this study.

These are (i) the Falcade region and (11) the Passo Rolle region, where the kunzi group (ssp.

kunzi) is in contact with the euryale group (represented by ssp. ocellaris), and (111) the Trafoi

valley and the Sulden valley upstream of their confluence near Gomagoi, where the kunzi group

(represented by ssp. pseudoadyte) meets the adyte group. The fourth exchange window, the

upper Valtellina (Adda valley), was not sampled.

Sampling

E. euryale has a two year life cycle. In the contact regions, samples were collected in 2009,

2011 and 2013, so the cohorts on the wing were the same in each collecting season. Samples

in the Falcade region are labelled F1-F6 (Fig. 2), in the Passo Rolle region RI-RS (Fig. 3),

and in the Trafoi region T1—T5 (Fig. 4). In the Passo Rolle region, special attention was paid

Figure 1. The Alps, with geographic boundaries of the taxa of E. euryale discussed in this paper. Light

grey: mountain areas above 1000 m, dark grey: lakes. Solid lines: group boundaries. Dotted lines: subspe-

cies boundaries. Circle — the adyte group, with ssp. adyte (1); squares — the euryale group with ssp. isarica

(2) and ssp. ocellaris (3); diamonds — the kunzi group, with ssp. pseudoadyte (4) and ssp. kunzi (5). T =

Trafoi test region, R = Passo Rolle test region, F = Falcade test region. The intergradation zone isarica /

ocellaris is included in the ocellaris area.

22 CupEDo: Reproductive isolation and intraspecific structure in Alpine populations of Erebia euryale...

to Passo Colbricon. Here, a short transect was sampled and treated as a separate sample set:

C1-C3 (Fig. 3).

Samples from the contact regions (called “test samples”) were compared to samples from

populations outside the contact region (“reference samples”). Each reference sample consists

of 150 individuals of one subspecies. These originate from five localities, scattered in the ter-

ritory, but at a distance of at least 40 km from the exchange regions. In the adyte territory, no

samples were taken west of Lago Maggiore, since ssp. adyte might not be genetically homoge-

neous in its entire distribution area (Schmitt and Haubrich 2008). For sampling locations and

sampling sizes see Figs 2-4 and Table 1.

Female genitalia of different groups are indistinguishable. Female wing pattern enables a

certain separation of the ssp. kunzi and ocellaris, but not of the ssp. adyte and pseudoadyte.

Therefore this study is entirely based on male characters.

Genital preparation

Male abdominal tips were macerated for 10 min in a 10% KOH solution at 100°C, the genital

apparatus was extracted, dehydrated in ethanol (96%) for 10 min, and embedded in euparal.

Variables

Individual males were characterised on the basis of four variables. Three of these are char-

acteristics of the valve and one is derived from the wing pattern. Valve characteristics were

measured on the right valve, as described and figured in Cupedo (2010): 1. shoulder index:

the height/width ratio of the dorsal shoulder; 2. first tooth: the relative position of the most

proximal tooth; 3. tooth length: the relative length of the longest tooth on the shoulder, as a

percentage of the valve length; 4. the presence or absence of discriminating elements in the

male wing pattern. A character is regarded discriminating if both its specificity and its positive

predictive value are > 0.90, according to the extensive dataset in Cupedo (2010). Discrimi-

nating elements are the following (see Table 2): for ssp. ocellaris brown ringed ocelli on the

hindwing underside; for ssp. kunzi white pupils in the ocelli on the forewing upperside, the

absence of ocelli on the forewing upperside or on the hindwing upperside, no traces of the

brown postdiscal band on the forewing upperside or on the forewing underside. In the case of

ssp. adyte and ssp. pseudoadyte, none of the characters met these criteria. Character 3, on the

other hand, does not discriminate between ssp. kunzi and ssp. ocellaris. Hence, in the contact

zone adyte-pseudoadyte, characterization of individuals in test populations was based on vari-

ables 1, 2 and 3, in the contact zone kunzi-ocellaris on variables 1, 2 and 4.

Measurements for shoulder index and first tooth were made using a Mitutoyo 176-902 meas-

uring microscope (magnification 30-fold). Tooth length was measured from calibrated micro-

photographs on a monitor (final magnification 1000-fold). Variable 4 was assessed with +2

dioptre glasses.

Nota Lepi. 37(1) 2014: 19-36

Scoring system and data analysis

Characterizing individuals and samples. A

scoring system was developed by which each

individual and each sample could be charac-

terised. For each variable, the values of all in-

dividuals in both reference groups (e.g. adyte

and pseudoadyte) were combined. The hereby

obtained numerical range was split into seven

categories, labelled -3 to +3. The centre of the

zero category of the scale coincides with the in-

tersection of the frequency distributions of the

two reference groups. For each male, the val-

ue of each variable was converted into a score,

equal to the category it falls into, thus ranging

from -3 to +3. Each individual male was char-

acterised by the sum of its scores for the three

variables, potentially ranging from -9 (the most

adyte-like individuals) to +9 (the most pseudo-

adyte-like ones). The scoring procedure was

essentially the same in the analysis of the kunzi

and ocellaris samples, except for the fact that

variable 3 was replaced by variable 4, which

has only three categories: -3 (ocellaris), +3

(kunzi) or zero (no discriminating wing char-

acter present). Each sample was characterised

by the frequency distribution of its individual

scores.

Identifying transitional samples. The fre-

quency distributions of the scores in test sam-

ples were compared with those in the reference

samples, using the Mann-Whitney U test. A test

sample was considered transitional if it differed

significantly (p < 0.05, two-sided) from both

reference samples.

Test for hybridization. The question

whether a transitional sample contains hybrids

requires recognition of hybridization, not nec-

essarily of hybrid individuals. Testing for hy-

bridization was based on the assumption that

hybrid butterflies are hardly ever equal to one

of their parents: they exhibit either a combi-

nation of parental characters, or they have in-

termediate characters (Mayr 1963). In either

Figures 2-4. Topography of the test regions. 2.

Falcade test region, 3. Passo Rolle test region and

4. Trafoi test region. Dotted: locations of test popu-

lations. Solid line in Figure 3: mountain chain. PR

= Passo Rolle; PC = Passo Colbricon. Note the di-

fferent scale in Figure 4. Reproduced from Tobacco

maps 022 (Figures 2 and 3) and 08 (Figure 4).

24 CupEpo: Reproductive isolation and intraspecific structure in Alpine populations of Erebia euryale...

Table 1. Sampling locations, sample codes and sample sizes of the sampled E. euryale populations. Code

= sample code used in this paper; N = sample size

Sample location Code Code N

ssp. adyte ad 150 Test region Falcade

Eggen am Simplon (CH) 30 | Falcade-1 F1 60

Pontresina (CH) 26 | Falcade-2 F2 35

Monte Tamaro (CH) 26 | Falcade-3 F3 60

Langtauferertal (1) 30 | Falcade-4 F4 48

Fusio (CH) 38 | Valle di Gares F5 14

ssp. pseudoadyte ps 150 | Valle di Gares F5 14

Val Malga, Adamello (1) 33 Passo San Pellegrino F6 15

Monte Baldo (I) 30 Test region Passo Rolle |

Monte Tremalzo (I) 30 | Paneveggio R1 | 22

Monte Legnone (1) 27 Passo Rolle road, west R2 29

Pradalago, Presanella (1) 30 | Sentiero laghi di Colbricon R3 44

ssp. kunzi ku 150 | Path Rolle - Colbricon | R3 50

Monte Cavallo (I) 47 | Passo Rolle road, east R4 30

Vette Feltrine (I) 30 San Martino di Castrozza R5 30

Cimonega (I) 30 | Passo Colbricon (north) C1 37

Col Visentin (I) 30 | Passo Colbricon C2 25

Monte Grappa (I) 13 | Passo Colbricon (south) C3 50

ssp. ocellaris oc 150 Test region Trafoi

Geissler Gruppe (1) 30 | Trafoi, Madatsch T1 60

Sesto (I) 30 | Trafoi, left bank T2 60

Plöckenpass (I) 30 | Trafoi, south of camping T3 39

Passo Fedaia (I) 30 | Sulden, south of Karnerbriicke T4 60

Lienzer Dolomites (A) 30 | Martelltal, Lify alm TS 60

case, the scores tend to drift to zero, i.e. towards lower absolute values. To test for such shift,

category labels were made absolute, and the values of identical categories were added (-9 and

+ 9 became 9, and the numbers in the categories -9 and +9 were added in category 9, etc.).

The cumulative graph of the values thus obtained (hereafter called the “absolute graph”)

was compared to the graph of an imaginary cohabitational population without hybridization,

created by combining both reference samples. If a test sample contains hybrids, its graph will

show a left shift when compared to the reference graph. The magnitude of such a shift is an

empirical estimation of the degree of hybridization in the transitional population. The proce-

dure is illustrated in Fig. 5.

Test for clinal variation. In each test sample, the proportions of individuals with a neg-

ative score and with a positive score were calculated. This provides a good approximation

of the proportions of parental characters in the population, without the need of individual

identification, and regardless of the hybridization rate in the population. These proportions

were used to detect geographic clines in the contact zones. Because of the small overlap of the

distributions of scores in the adyte and pseudoadyte reference samples, 3.3% of the positive

scores are incorrectly classified as pseudoadyte, and 5.3% of the negative scores are incorrect-

ly classified as adyte. The more one of both types is predominant, the more this will affect the

Nota Lepi. 37(1) 2014: 19-36

Table 2. Specificity (sp) and positive predictive value (ppv) of characters discriminating between the

subspecies ocellaris and kunzi of E. euryale. * —Data underlying Table 4 in Cupedo (2010). Abbrevia-

tions: Hw = hindwing; Fw = forewing; Up = upperside; Un = underside; Oc = apical ocelli; B = brown

postdiscal band.

ssp character | value #true* | # false* nn en En no

ocellaris | HwUnOc | with brown ring | pos 314 7 0.9857 0.9708-0.9940

| neg 54 484 0.9782 0.9556-0.9911

kunzi FwUpOc with white pupil | pos 183 3 0.9918 0.9763-0.9982

| neg 308 365 0.9839 0.9535-0.9965

kunzi FwUpOc absent pos 78 7 B 0.9810 0.9612-0.9923

neg 413 361 0.9176 0.8376-0.9661

kunzi HwUpOc _ | absent pos 323 30 0.9158 0.8857-0.9443

neg 167 338 0.9150 0.8809-0.9419

kunzi FwUpB |absent pos 157 4 0.9891 0.9724-0.9970

neg 334 364 0.9752 0.9375-0.9930

kunzi FwUnB absent pos 115 0 | 1.0000 0.9899-1.0000

neg 376 368 1.0000 | 0.9681-1.0000

adyte / pseudoadyte ratio. As a result, any clinal character gradient in the adyte / pseudoadyte

contact zone will be slightly underestimated.

Statistics

Statistical tests were performed with the SPSS 12.0 package. Specificity and positive predictive

value of discriminating characters were calculated with MedCalc online statistical calculators

for Windows, version 12.7.8.

Material deposition

Samples and genital preparations are deposited in the collection of the author.

Results

Identifying transitional samples

The distributions of individual scores per sample are shown in Table 3. The results of a statistic

comparison of these distributions (Mann Whitney test), with each other and with the reference

samples, for each of the contact zones, are presented in Fig. 6. In each contact zone there were (i)

test samples that did not differ significantly either from each other or from one of the reference

samples, but did differ from the second reference sample (grey in Fig. 6), and (ii) test samples that

differed significantly from both reference samples and from the test samples mentioned under (i).

The latter were considered transitional. These were: F2, F3 and F4 in the Falcade region; R3, Cl

and C2 in the Passo Rolle region and T3 and T4 in the Trafoi region.

26 Cupebo: Reproductive isolation and intraspecific structure in Alpine populations of Erebia euryale...

(| Ref 1 = Ref2 : MRefi+Ref2 MTest samp

Ref 1+Ref2 Test sample

7 A TR Te

-9 -6 =? =6 -5 -4 -3 -2.51 0 1 2456 7 8.9

Figure 5. Illustration of the procedure for detection of hybrids. A: frequency distributions of scores in

reference samples; B: frequency distribution of scores in the combined reference samples, and in a test

sample; C: as B, X-axis categories are made absolute. D: as C, cumulative. X-axis: score (A and B) or

absolute score (C and D); Y-axis: proportion of the sample.

Test for hybridization in transitional samples

In Fig. 7, the absolute graphs of all samples are compared to the pooled reference samples. A

left shift is obvious in all transitional samples and, to a much lower degree, in some samples

that are not classified as transitional. This implies that intergroup mating produced adult hy-

brıds in each of the studied contact zones.

The composition of the transitional samples

The score distributions of the transitional samples in the hybrid zones kunzi / ocellaris are differ-

ent from those in the hybrid zone adyte / pseudoadyte (Fig. 8). In the former, the ranges almost

cover the combined ranges of both reference samples. F2, R3 and C2 show an explicit bimodal

distribution; in F3, the distribution is strongly biased towards the kunzi phenotype. In the contact

area adyte / pseudoadyte, on the other hand, the range of the individual scores covers no more

(T3) or hardly more (T4) than the range of one of the reference samples. Typical pseudoadyte in-

dividuals are lacking in T3, and typical adyte are absent from T4. The distributions are unimodal,

but both are skewed towards the hybrid end of the scale.

Test for clinal variation in the transition zones

The fractions of negative and positive scores (Table 4) exhibited a clinal gradient in each of the

contact zones. In the Falcade region, this is along the (F6-F 1 )-F2-F3-F5 line; in the Passo Rolle

region along the RI-R2-R3-(R4-R5) line and along C1-C2-C3, and in the Trafoi test region

Nota Lepi. 37(1) 2014: 19-36 97

Table 3. Distributions of individual scores per sample, ın reference samples and test samples of E. eu-

ryale.

|| -9 | -8 | -7 | -6 SE EN

Reference | ad| | 1 | 2 | 11 | 12 | 23] 34/36/21| 5 | 4/1 150

samples | ps | 3 | 5 | 11] 31 | 33 150

oc 21918 a0 255) ous 150

ku 4] ı 2 | ı | 150

Falcade | Fi HAINE 1 60

F2 Aree E in BEIEIEIEIFREIE 35

F3 2 AEIEZEITEENITREITIE 60

F4 je Irre 48

FS | | Axe ® TE 14

F6 SETZIETE 15

Passo 2.1 6408,12. 3 1 22

Rolle R2 | 1 SEFPIRIEIE 29

R3 Deere 50

R4 2] 1 me 2 30

R5 2 30

C1 eae 218 RAP 37

C2 2 Er gese 4 25

C3 ae ARE”

Trafoi T1 Daran 4 60

T2 64 at i a eS: sut 60

T3 Aa 415 39

T4 ail APTE UE TE 60

TS DÉFIER Pi 60

along (T1-T2)-T3-T4-T5. Samples in brackets do not differ significantly from each other (p <

0.05, two-sided).

Discussion

The composition of the hybrid samples

Transitional samples were present in each of the three contact zones, and in each of them hy-

bridization took place, so the contact zones discussed in this paper explicitly are hybrid zones.

There is, however, a noticeable variation in the score distributions of the test samples, both

among and within contact areas. Field observations yield enough additional information to

explain these differences.

I. The Falcade test region. In the Falcade contact zone, the northern slopes of the Valle

di Vales are inhabited by ssp. ocellaris, whereas ssp. kunzi occupies the southern slopes.

Samples F1 and F3 were taken from high-density populations, living in clearings in a mixed

fir-larix forest, with F1 at the upper tree limit on the northern slope (1870 m), and F3 in the

lower part of the opposite slope (1670 m). In between, individuals of E. euryale were scarce,

flying along roadsides and on hay meadows. A local concentration was only found at location

28 Cupebo: Reproductive isolation and intraspecific structure in Alpine populations of Erebia euryale...

Ai ee ETES.

| ad . =

13 a " lca

| | | | BE

el) | |

ps | | | JON

MOCÈC

>| |

Figure 6. Mann-Whitney’s significance levels for pair wise comparison of the frequency distributions of

the scores of all samples. A: Falcade test region; B: Trafoi test region; C: Passo Rolle test region; D: Passo

Colbricon. For sample codes see Table 1. Legend for p values: * p < 0.05, ** p < 0.01, *** p < 0.001, n-s.

not significant.

F2. The contact zone, which was sampled in 2009, was revisited in 2013. The situation at F1

and F3 was unaltered. In between, though, more individuals were present than in 2009, but

no concentration was observed at site F2. This gives the impression of two stable, permanent

populations (F1 and F3), from where individuals swarm out, annually, into the less suitable

zone in between. This would explain the temporary character of F2, as well as its bimod-

al distribution. The score distribution of F1 does not differ significantly from the ocellaris

reference sample, and F3 shows a kunzi-like distribution, which tails out on the left side,

evidencing hybridization with ocellaris. Apparently, down slope roaming (F1 — F3) exceeds

uphill movement in this locality.

One kilometre downstream, the valley floor (at 1250 m) was inhabited by the hybrid popu-

lation F4. Due to the inaccessibility of the slopes here, no migration was actually observed, but

the composition of the population strongly supports a regular influx from both sides. Given the

Nota Lepi. 37(1) 2014: 19-36 29

ere et |

— Sample F1 _ — Sample R1 0.2 2 4a — Sample T1

— ocellaris + kunzi | | — ocellaris + kunzi — adyte + pseudoadyte

I 0.0 RTE ar ok 1 TI ree RES DDS |

4 5 6 7 8 9

— a)

Tea _—

= Sample F2 — Sample R2 — Sample T2

— ocellaris + kunzi — ocellaris + kunzi — adyte + pseudoadyte | |

| r = i 0.0 a TRS =] = TEEN = |

! |

4 5 6 7 8 9 o 1 2 3 4 5 6 7 8 9

= = A NENNE ee ee ee a

| 1.0

| — Sample F3 — Sample R3 io — Sample T3 |

— ocellaris + kunzi — ocellaris + kunzi — adyte + pseudoadyte

1 RERO EISEN | 0.0 ! 1 | CS ERS TER EE

4 5 6 7 #8 9

0 19273 4.50 7 8 à

elle

|— Sample F4 — Sample R4 02 - 2 — Sample T4 |

| ee — ocellaris + kunzi — ocellaris + kunzi | — adyte + pseudoadyte | |

1 | 0.0 r 1 re (AGE A RS L £ (oS SS 1 T CE) |

OMAN 2 3 4 5 6 7 8 9 0 1 2 3 4 5 6 7 8 9 | O°, 2053 AM 596 7.487029

1.0

— Sample T5

— adyte + pseudoadyt

— Sample F5

— ocellaris + kunzi

— Sample R5 02

— ocellaris + kunzi

Os ein 2.3 4 5 6 7 +8

— Sample C2

— ocellaris + kunzi

— Sample F6

— ocellaris + kunzi

je}

— Sample C3

— ocellaris + kunzi

| _ — Sample C1

| — ocellaris + kunzi

5 6 7 8 9

Figure 7. Test for the presence of hybrids in test samples of E. euryale in contact zones between groups.

X-axis: absolute score; Y-axis: proportion of the sample. For sample codes see Table 1.

30 Cupebo: Reproductive isolation and intraspecific structure in Alpine populations of Erebia euryale...

ocellaris @ kunzi

a ENIAC NON AN ENS TA

Sample F2

D FRE H

B Sample F3

en NN

Sample F4

“9 “Bont, 6.251, 24. ES e290 10) 51,025 3 745260700829 -9 -8 -7 -6 -5 -4 -3 -2 -1 0 1 2 3 45 6 7 8 9

Figure 8. Frequency distribution of individual scores in reference samples (A, F) and in transitional test

samples of E. euryale. X-axis: score; Y-axis: proportion of the sample. For sample codes see Table 1.

Nota Lepi. 37(1) 2014: 19-36 +

Table 4. Proportions of individuals with a negative and with a positive score, in each of the £. euryale sam-

ples.

lad oe | ku RI1| R2| R| R RB ca C2] ©

<0| 0.93] 0.05| 1.00] 0.00 1.00] 0.97| 0.72] 0.00] 0.00] 0.59} 0.36] 0.00

>0| 0.03] 0.87) 0.00] 1.00 0.00} 0.03] 0.24] 1.00] 1.00] 0.30] 0.64] 1.00

Falcade Trafoi

Scores| F1] PR F3| F4] F5| F6 T1] T2] T3] T4) TS

<0| 097| 0.46] 0.08) 0.27| 0.00| 0.93 0.83] 0.90! 0.67] 0.22| 0.07

>0| 0.02] 0.54] 0.88] 0.63] 1.00] 0.00 0.07| 0.05] 0.23] 0.62! 0.80

high population density in a favourable habitat, this population has to be considered a perma-

nent and breeding population. This might explain why the hybrid ratio 1s the highest among the

analysed hybrid populations.

2. The Passo Rolle test region. In the Passo Rolle region, it is the Latemar chain and its

continuation, the Focobon chain, that separate kunzi from ocellaris. Two depressions in

this chain, the Passo Rolle and the Passo Colbricon, are potential exchange windows. The

largest one, Passo Rolle, is an ecologically devastated area, which offers no suitable habitat

to E. euryale. A single specimen was observed. Exchange of individuals takes place over

the much narrower Passo Colbricon, 2.5 km southwest of Passo Rolle. Here, E. euryale

was present in relatively high density, on the pass and on both sides. In this continuous

population, connecting the ocellaris area with the kunzi area, an extra set of three samples

was taken from nearby sites. C2 was collected on the pass (within 20 metres around the

pass mark, Fig. 9), Cl 200 to 350 m to the North of the pass and C3 between 200 and 400

m from the pass at its southern slope. This southern sample did not differ significantly from

the kunzi reference sample, nor was there any detectable hybridization (Table 3, Fig. 7). On

the pass, the ocellaris portion was 36%, in the more northern C1 sample it was already 59%.

This justifies three conclusions: the Colbricon pass, despite its small size, is an important

exchange corridor between kunzi and ocellaris; dispersal is predominantly northward, as

Cl and C2 contained a substantial portion of kunzi, but ocellaris was absent from C3; and

maintenance of this sharp separation within a continuous population requires a strong repro-

ductive isolation mechanism.

3. The Trafoi test region. In the Trafoi contact zone, adyte is widespread west of the Trafoi

valley, and pseudoadyte occurs east of the Sulden valley. The Tabaretta chain of the Ortler Mas-

sif is inserted in between (Fig. 4). The hybrid populations T3 and T4 were found at its foot. The

distance T3-T4 is 4.5 km in a straight line (across the Tabaretta ridge, 2800 m), and 8 km when

measured along the 2000 m contour line (the most probable migration path). Both distances by

far exceed the mean individual range of flight. Gene exchange therefore might take several gen-

erations, so parental individuals from T3 will normally not reach T4 and vice versa. However,

the influx of hybrid phenotypes biases the score distributions of both populations towards the

hybrid end of the scale (compare Figs 8G and 8H to Fig. 8F).

32 Cupebo: Reproductive isolation and intraspecific structure in Alpine populations of Erebia euryale...

Figure 9. Passo Colbricon (1908 m), an important exchange corridor of ssp. kunzi and ssp. ocellaris. View

to the south (31.vii.2013).

Clinal gradients in hybrid zones

In each of the contact zones, a clinal gradient of characters is obvious (Table 4). Because mor-

phologic and genetic clines have been shown to be coincident and concordant (Barton & Hewitt

1985; Collins et al. 1993; Dasmahapatra et al. 2002), the morphologic gradient can be consid-

ered to reflect genetic intergradation. Consequently, the rate of introgression can be estimated

from the steepness of the morphological gradient.

The kunzi group and the euryale group. In the Falcade contact area, the fraction of ocellaris

characters drops from 0.97 to 0.08 between F1 and F3, and the fraction of kunzi characters

from 0.88 to 0.02 in the opposite direction (Table 4). That is a mean decline of 87.5% across

one kilometre. In the Passo Rolle region, it is 97% across two kilometres (between R2 and R4),

and on the Colbricon pass it is 65% over 400 m. These declines are extremely steep, especially

when considering that the distances between adjacent samples in all cases are within the normal

individual range of flight. In both contact areas, explicit hybrid samples exist (F2, R3, C1, C2),

but the more peripheral samples are hardly affected. Morphologically detectable introgression

fades out within two kilometres.

The kunzi group and the adyte group. In the Trafoi region, the mean decline is 42% over at

least 4.5 km, more probably 8 km. Due to the different spacing of sampling, these data cannot

directly be compared to those from the kunzi / ocellaris contact regions. Nonetheless, if either

Nota Lepi. 37(1) 2014: 19-36 33

in the Falcade or in the Passo Rolle region two populations had been analysed 4.5 km apart,

hardly any morphological evidence of introgression was to be expected. In the Trafoi region,

though, the decline over this distance is only 42%. This at least justifies the conclusion that

introgression between the kunzi group and the adyte group is less inhibited by reproductive

barriers than between the kunzi group and the euryale group.

The euryale group and the adyte group. Rezbanyai-Reser (1991) described three contact

sites of the euryale group (ssp. isarica) and the adyte group (ssp. adyte) in Switzerland: Has-

liberg (BE), Gitschen (UR) and Rophaien (UR). His observations are based on wing characters

only, and quantitative data are lacking. In each of these localities, he found a transition zone

less than 100 m in width. Intermediate individuals flew together with both parental forms, in the

absence of any natural barrier. This is fully comparable to the situation in the Falcade and Passo

Rolle regions. In one of his localities, Rezbanyai-Reser (1991) made the important observation

that the situation remained unaltered over nine years. In other words, no progress of introgres-

sion was observed. Hybrid individuals seemed to be generated de novo in each generation.

Sonderegger (2005) described two comparable situations in Switzerland, based on both wing

pattern and valve characters. At Klosters (GR), a cohabitation zone of about 1 km was found.

Only at one specific site in this range, intermediate individuals were found, flying together with

both parental forms. At Monstein (GR), a cohabitation site without intermediates was observed.

An entirely different situation was recorded in two other sites (Brusons, VS and Grindelwald,

BE). Here, the ssp. isarica and ssp. adyte areas are spatially separated, and Sonderegger found

an intermediate population in between. According to Mayr (1942), this is a recurrent phenom-

enon when hybrid populations are deprived of parental influx over a long period of time. Mayr

(1942) states that these populations may achieve phenotypic stability by continuous selection

against the most unbalanced hybrid genomes.

Subspecies within groups. Hybrid zones of two subspecies of E. euryale belonging to the

same group are rare, since most of them have allopatric distributions. It is only in the Pyrenees

and in the Alps that two subspecies of the same group (the euryale group) are in secondary con-

tact. In both cases, one of the two subspecies is strongly melanistic, which enables easy identi-

fication of hybrid individuals by wing pattern. The hybrid zone in the Pyrenees is insufficiently

documented, but it covers a considerable part of the Pyrenees (pers. obs.). The hybrid zone in

the Alps (ssp. isarica and ssp. ocellaris) has been mapped (Cupedo 2010). All populations in

the intergradation zone mainly consist of hybrid individuals; parental types are rare or absent.

The hybrid zone attains its maximum width of about 40 km in the eastern Alps. More important

than its actual width is the fact that, in contrast to the hybrid zones between groups, introgres-

sion between these intra-group subspecies proceeded until further dispersal was inhibited by

natural barriers (mountain chains in the north and river valleys in the south). Obviously there

is a discrepancy between the narrow but stable hybrid zones between groups, and the freely

expanding hybrid zone between within-group subspecies.

The cause of the difference between hybrid zones

More or less stable transition zones between genetically distinct populations have been de-

scribed in a great variety of organisms (Mayr 1963; Barton & Hewitt 1985; Jiggins & Mal-

let 2000; Arnold 2006; Schmitt & Miiller 2007; Schmitt et al. 2007). They range from some

34 Cupebo: Reproductive isolation and intraspecific structure in Alpine populations of Erebia euryale...

hundreds of metres to some hundreds of kilometres. Barton and Hewitt (1985) showed that

the width of hybrid zones strongly depends on the balance of two antagonistic factors: dis-

persal and selection. Immigration into the region of overlap tends to widen the transition zone

but is continuously counteracted by selection. This selection may be either ecological, i.e. by

differences in the environment or by different adaptations to the environment, or genetic, by

selection against recombinant genotypes (Barton & Hewitt 1985; Arnold 2006). In the contact

sites studied in this paper, ecological factors can be ruled out as selecting factors. The habitats

at both sides of the intergradation zones are largely identical, and there are no indications of

different ecological preferences among Alpine subspecies of E. euryale. Consequently, genetic

selection, i.e. a decreased genomic compatibility of the populations in contact, is the most like-

ly factor determining the width of the hybrid zones in E. euryale. This selection may be pre-zy-

gotic, by assortative mating, or post-zygotic, by reduced hybrid fertility or viability. In the case

of E. euryale, the occurrence of hybrids is a sign of random or at least incompletely assortative

mating, and the low introgression rate is an indication of reduced hybrid fertility or viability.

Considering the width of the introgression zone, we should keep in mind that morphological

markers are far less sensitive than genetic ones. Barton and Hewitt (1985) showed that gradi-

ents in secondary contact zones are s-shaped, and that introgression tails out in both directions.

Morphologically, these tails will remain hidden because of a lack of resolution. In general, the

width of the intergradation zone will be underestimated when based on morphology alone. Gei-

ger and Rezbanyai (1982) have already demonstrated this phenomenon. They found a signifi-

cantly lower genetic distance (Nei 1972) between adyte and the isarica population at Hasliberg

(one of the Swiss contact sites) than between the same adyte population and a more remote, but

morphologically identical isarica population (D=0.036 and 0.073 respectively!). This demon-

strates gene flow by introgression, which could not be detected morphologically.

Differentiation levels

This study of hybrid zones reveals that strong reproductive barriers exist between the euryale

group and both the adyte group and the kunzi group. Our results suggest a less strong repro-

ductive isolation of the adyte group and the kunzi group, but the different spacing of the test

samples and the different characters used to discriminate between the groups impede an unam-

biguous numeric comparison of the results. Reproductive barriers between the subspecies is-

arica and ocellaris, both belonging to the euryale group, are so weak that they suggest random

mating and a high hybrid viability. Consequently, at least two, maybe three, hierarchical levels

of reproductive isolation exist between E. euryale populations. Since the degree of reproduc-

tive isolation is positively correlated with genetic distance, i.e. the duration of the interruption

of gene flow (Coyne & Orr 1997; Jiggins & Mallet 2000), the results support the idea of at least

two, maybe three, temporally separated differentiation events. In the latter case, disjunction of

the euryale group and the adyte-kunzi precursor would have preceded the disjunction of the

adyte group and the kunzi group. This scenario would be in agreement with the morphological

structure, as the adyte group and the kunzi group are more similar to each other, both in male

genital features and in wing pattern, than either is to the euryale group.

Nota Lepi. 37(1) 2014: 19-36 35

Acknowledgements

I am indebted to Prof. Dr. Jan E.R. Frijters, who developed the scoring system and the hybridization test,

to Dr. Tamara van Mölken for her critical remarks on the initial version of manuscript, to Prof. Dr. Thomas

Schmitt for acting as the editor, and to Mr. Hub L.E. Peters who corrected the English text.

References

Arnold ML (2006) Evolution through genetic exchange. Oxford University Press, Oxford. 252 pp.

Barton NH, Hewitt GM (1985) Analysis of hybrid zones. Annual Review of Ecology, Evolution, and

Systematics 16: 113-148.

Collins MM, Britten HB, Rivers V (1993) Allozyme analysis of a known hybrid zone between Hyalo-

phora euryalus and H. columbia gloveri (Lepidoptera: Saturniidae) in the California Sierra Nevada.

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Comes P, Kadereit JW (1998) The effect of quaternary climatic changes on plant distribution and evolu-

tion. Trends in Plant Science 3: 432-438.

Coyne JA, Orr HA (1997) Patterns of speciation in Drosophila revisited. Evolution 51: 295-303.

Cupedo F (2010) A revision of the infraspecific structure of Erebia euryale (Esper, 1805) (Nymphalidae;

Satyrinae). Nota Lepidopterologica 33: 85-106.

Dasmahapatra KK, Blum MJ, Aiello A, Hackwell S, Davies N, Bermingham EP, Mallet J (2002) Infer-

ences from a rapidly moving hybrid zone. Evolution 56: 741-753.

Geiger H, Rezbanyai L (1982) Enzymelektrophoretische Untersuchungen tiber die Verwandtschaftsbezie-

hungen bei Erebia, mit besonderer Berücksichtigung der Taxa euryale isarica Rühl und adyte Hübner

(Lep.: Satyridae). Entomologische Zeitschrift 92: 49-63.

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mountain plant Anthyllis montana L. (Fabaceae) inferred from ITS sequences and amplified fragment

length polymorphism markers. Molecular Ecology 11: 447-463.

Mayr E (1942) Systematics and the origin of species. Columbia University Press, New York. xiv+334 pp.

Mayr E (1963) Animal Species and Evolution. Belknap Press, Cambridge, MA. 797 pp.

Nei M (1972) Genetic distance between populations. American Naturalist 106: 949.

Rezbanyai-Reser L (1991) Die drei Zentralschweizer Kontaktstellen der Erebia euryale - Unterarten isar-

ica Heyne und adyte Hbn. (Lep., Satyridae). Entomologische Berichte (Luzern) 25: 77-90.

Schmitt T (2007) Molecular biogeography of Europe: Pleistocene cycles and Postglacial trends. Frontiers

in Zoology 4: 11.

Schmitt T (2009) Biogeographical and evolutionary importance of the European high mountain systems.

Frontiers in Zoology 6.

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of the butterfly Erebia medusa (Satyrinae, lepidoptera) in Central Europe. Journal of Zoological Sys-

tematics and Evolutionary Research 45: 39-46.

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of Zoological Systematics and Evolutionary Research 49: 119-132.

Nota Lepi. 37(1) 2014: 37-42 | DOI 10.3897/n1.37.7954

Scotopteryx kurmanjiana, a new species from the Kopet-Dagh

Mountains (Lepidoptera, Geometridae, Larentiinae)

HOSSEIN RAJAEI SH.', GYULA M. LASZLO?

1 Biocentre Grindel and Zoological Museum, University of Hamburg, Martin-Luther-King-Platz 3, 20146

Hamburg, Germany; Stuttgart State Museum of Natural History, Rosenstein 1, 70191 Stuttgart; hossein.rajaei.

shoorcheh@uni-hamburg.de

2 Fadrusz u. 25, H-1114 Budapest, Hungary

http://zoobank.org/E9C4F 2AA-0CA 7-4147-BSE2-B748A38F 2154

Received 19 November 2013; accepted 10 March 2014; published: 15 June 2014

Subject Editor: Sven Erlacher

Abstract. Scotopteryx kurmanjiana sp. n. is described from the Kopet-Dagh Mountains in Northeast Iran

and South Turkmenistan. The new species is considered to be closely related to Scotopteryx kuznetzovi

(Wardikian, 1957) described from Armenia. Detailed description of the external morphology and the ge-

nitalia characters of the male with six colour photographs and two illustrations of genitalia are provided.

Introduction

Scotopteryx Hiibner, 1825, is a widely distributed genus occurring from North Africa through-

out Europe to the Pacific East Asia, South Africa and Southern America (Parsons et al. 1999;

Scoble and Hausmann 2007). Traditionally it belongs to the tribe Xanthorhoini of the subfamily

Larentiinae (Pierce 1914), but recently it was placed again into the tribe Scotopterygini Warren,

1895 (Viidalepp 2011; Schmidt 2013). The genus is one of the species-richest larentiine clades,

as more than 70 species are known worldwide (Scoble & Hausmann 2007).

In the course of a recent taxonomic revision of the subfamily Larentiinae (Rajaei Sh.

2012) and after re-describing and publishing the new records of the poorly known Scotop-

teryx kuznetzovi (Wardikian, 1957) (Rajaei Sh. and Stiining 2012), two peculiar Scotopteryx

kuznetzovi-like specimens have been discovered in the Zoological State Collection of Bavarian

State and the private collection of Gyula M. Laszlo, collected in the Iranian and Turkmenian

sides of the Kopet-Dagh Mountains. Although these two specimens are undeniably closely

related to S. kuznetzovi, their wing pattern and the genitalic characters show remarkable dif-

ferences in comparison to those of S. kuznetzovi. In addition, the Kopet-Dagh, where the two

specimens were collected, is about 1000 km from the known range of S. kuznetzovi, which is

distributed in South Armenia, West Iran and East Turkey, supporting the taxonomic separation

of the examined specimens. These observations led the authors to a description of a new species

Scotopteryx kurmanjiana sp. n. from the specimens from the Kopet-Dagh Mts as a potential

sister species of S. kuznetzovi.

38 RAJAEI & LASZLO: Scotopteryx kurmanjiana, a new species from the Kopet-Dagh Mountains

Methods

Genitalia of the examined specimens were dissected following standard procedures (Robinson

1976). Permanent genitalia slides were studied and photographed with a digital stereo-micro-

scope (ZEISS-SteREO: Discovery. V20) in ZFMK. The specimens examined in this study are

deposited in the following collections (acronyms after Evenhuis and Samuelson 2007): ZFMK

— Zoologisches Forschungsmuseum Alexander Koenig, Bonn, Germany; ZSM — Zoologische

Staatssammlung Miinchen, Germany; HMIM — Hayk Mirzayans Insect Museum, Tehran, Iran;

LG — number of genitalia slide made by Gy. M. Laszlo.

Scotopteryx kuznetzovi (Wardikian, 1957)

Figs 3, 4, 6

Material. Iran: 2 3: Basmendj [NW Iran, SE Tabriz], 15.x. [19]74, [leg.] Damanabi, gen. preps slide Nos: 1063 &

1064/2010 H. Rajaei (coll. HMIM). 3 3, Prov. Azerbayejan, E-Garbi, 11 km S of Shoet, 1350m, 31.x-01.xi.2003, leg.

P. Gyulai & A. Garai, slide No.: LG 2834; 1 4, Prov. Esfahan, 2750m, C-Zagros, Golestan Khuh, 10 km S of Khansar,

10-11.x.2001, leg. P. Gyulai & A. Garai, slide No.: LG 2339; 10 3, same data, but collected at 27-28.x.2003; 1 4, Prov.

Esfahan, 7 km NW of Natanz, (to Kashan), Kuh-e-Karkas, 1500m, 18-19.x.2003, leg. P. Gyulai & A. Garai; 5 3, Prov.

Esfahan, C-Zagros, 2600m, 2 km NE of Semirom, 20-21.x.2003, leg. P. Gyulai & A. Garai, slide Nos: LG 2832, 2833

(coll. Gy. M. Läszlö). Turkey: 2 À, Ost Türkei, Van, 2700 m, Güzeldere Pass, 28.ix.1981, leg. P. Kuhna, gen. prep.

slide No.: 3074, P. Kuhna; 1 4: Prov. Bitlis, Van Gölü, 19 km E of Ahlat, 1700 m, 42°34‘ E, 38°46‘ N, 18.x.1993,

leg. Gy. Fabian, B. Herczig, Gy. Läszlé and K. Szeöke, (coll. ZFMK). 4 à, Prov. Bitlis, Van Gölü, 19 km E of Ahlat,

1700 m, 42°34’ E, 38°46’ N, 18.x.1993, leg. Gy. Fabian, B. Herczig, Gy. Laszlo and K. Szeôke, slide No. 394 Gy. M.

Laszlo; 1 3, Prov. Dogubayazit, Ishak Pasha Serayi near Dogubayazit, 14-15.x.2003, leg. P. Gyulai & A. Garai (coll.

Gy. M. Laszlo).

Remarks. A large series of Scotopteryx kuznetzovi specimens was examined in order to

determine the individual variability of morphological characters within this species. As a sup-

plementary result of this survey, S. kuznetzovi has been recorded from several new localities,

extending our knowledge of its distribution.

Scotopteryx kurmanjiana sp. n.

http://zoobank.org/ACDC8372-F2A6-445 A-BCFF-B555558076E 1

Figs 1, 2, 5

Material. Holotype €, Turkmenistan, SW Kopetdagh, Garrygala env. 1994.xi.15-30, Leg. Miatleuski J., slide No.:

LG 1747 (ZSM). Paratype: 1 4, Iran, Prov. Khorasan, Kopet-Dagh Mts, 80 km NE of Qucan [Quchan], 1900 m,

37°28° N, 58°34° E, 30.x.2000, leg. B. Benedek & Gy. Fabian, slide No. LG 1746 (coll. Gy. M. Laszlo).

Description. (Figs 1, 2). Male: Wingspan 28-33 mm (Length of forewing: 16-18 mm).

Antenna bipectinate from base to tip, except 2—3 distal segments, rami moderately long, black,

dorsally unscaled, arising ventrally from the proximal end of the flagellum segments. Head,

thorax and abdomen covered with mixed brown-white scales. Frons broad, slightly protruding,

lower part smoothly covered with very small dark brown scales, upper margin covered with

mix of whitish and pale brown scales; vertex with large white and pale brown scales. Chaeto-

Nota Lepi. 37(1) 2014: 37-42 39

Figures 1-4. Wing pattern of adults (males). 1-2: Scotopteryx kurmanjiana sp. n. 1. Holotype; NE

Quchan, Iran; 2. Paratype; Garrygala, S Turkmenistan; 3-4: S. kuznetzovi. 3. Guzeldere Pass, E. Turkey;

4. Basmendj, NW Iran; a: dorsal view, b: ventral view.

semata transversally extended. Palpi short and narrow, acute at tip, just reaching beyond the

clypeus. Haustellum almost completely reduced. Index of spurs: 0-2-4. Forewings elongated,

apex and tornus rounded; apical patch absent; ground colour pale greyish-brown; basal area

and medial band slightly darker, edged with dark brown. Basal line indented. Antemedial line

roundly curved in the middle, with two moderately acute incisions. Medial line shadow-like,

poorly visible, median area uniformly pale greyish-brown, discal spot small, blackish, shar-

ply defined. Postmedial line wavy, roundly curved outwards in the middle. Submarginal line

blurred, poorly visible. Hindwings oval, elongated, cream-brown, crossline absent, discal spot

very small, poorly visible. Fringes in both wings unicolorous and consisting of a row of shorter

and darker basal scales and a row of longer and lighter terminal scales. Underside pale greyish

40 RAJAEI & LASZLO: Scotopteryx kurmanjiana, a new species from the Kopet-Dagh Mountains

brown, generally paler than upperside, basal area and middle band darker, patterns of upper

side partly visible. Underside of the hindwing even paler than that of the forewing. Abdomen

long, narrow, light grey. Coremata absent.

Male genitalia (Fig. 5). Vinculum rather short and broad, gently rounded. Valva short, broad

at the base, sacculus well sclerotized, slightly arched, having conspicuous, trapezoidal apical

lobe. Distal margin of valva broadly rounded, setose; costal margin strongly sclerotized, slight-

ly arcuate, with an apically rounded, finger-like apical process surpassing the distal edge of val-

va. Transtilla present, curved, band-like. Uncus broad at base, triangular with narrow, acute tip,

curved ventrad. Anellus lobes conically elongated, distally rounded, setose. Juxta X-shaped.

“

’

“

Figures 5-6. Male genitalia. 5. Scotopteryx kurmanjiana sp. n., Holotype (Slide No.: LG 1747 M; S

Turkmenistan); 6. S. kuznetzovi (Slide No.: 1063 H. Rajaei; NW Iran); a: genital armature, b: phallus.

Abbreviations: an.lob. Anellus lobe; cos. Costa; tr. Transtilla; unc. Uncus; v. Valva.

Nota Lepi. 37(1) 2014: 37-42 4]

Phallus tubular, slightly longer than valva, gently curved, distal half covered with the densely

spined part of manica, apically with a well-developed finger-like process of carina; vesica bears

a bunch (over 10) of broad-based, strongly sclerotized cornuti.

Female. Unknown.

Diagnosis (Figs 1-6). According to its morphological and genital features, Scotopteryx kur-

manjiana appears to be closely related to S. kuznetzovi but is nevertheless easily distinguishable

by several characters (for the distinctive features between S. kuznetzovi and other related Scoto-

pteryx species see Rajaei Sh. and Stüning 2012). External features: the new species has less

protruded frons in comparison with S. kuznetzovi and in general less shiny wing surface; the

transverse lines are much less sharply defined, lacking the white highlight, which is very cha-

racteristic in S. kuznetzovi; the median area is much paler, less contrasting in S. kurmanjiana;

the submarginal line is inconspicuous, more or less shadow-like, whereas it is sharply defined

with whitish scales in S. kuznetzovi; the discal spots are present in both wings while these are

absent in the hindwings of S. kuznetzovi. The apical patch of S. kurmanjiana is indistinct while

a whitish triangular patch is present in S. kuznetzovi; the fringes are uniformly whitish-grey,

while they are chequered with dark brown in S. kuznetzovi. Finally, the hindwing of S. kurman-

Jiana is unicolorous, without transverse line or band, while a well-defined dark-grey medial

band is present in S. kuznetzovi.

The specific differences between the two species are well expressed in genitalia (see the Figs

5a and 6a): the new species has shorter and smaller apical lobe of sacculus, more sclerotized,

much thinner costal margin of valva with conspicuously shorter and apically more tapering cos-

tal process, broader base of uncus, somewhat more strongly sclerotized and differently shaped

juxta. The distal half of the phallus of the new species is covered with densely spined part of

manica, while the spinose part of manica is only one third of the length of the phallus in S.

kuznetzovi. Finally, the vesica of S. kurmanjiana is armed by more than 10 well-sclerotized

cornuti, while the number of cornuti in S. kuznetzovi is fewer than five.

Bionomy. The known specimens of S. kurmanjiana were collected in the late autumn, sim-

ilarly to S. kuznetzovi. The foodplant and the early stages are unknown.

Distribution. The species is known to occur on both sides of Kopet-Dagh Mountains in NE

Iran and S Turkmenistan.

Etymology. The name of the species is dedicated to the Kurmanj people in northeastern

Iran. This Kurdish tribe migrated from Kurdistan and settled in Khorasan-e Shomali, mainly in

Quchan, Shirvan and Bojnurd.

Acknowledgements

The authors are indebted to Dr. Axel Hausmann (Munich), Dr. Dieter Stiining (Bonn) and Dr. Helen Al-

ipanah (Tehran) for their kind loan of material. We are also grateful to Dr. Läszlö Ronkay for his useful

advices during the preparation of the manuscript.

References

Evenhuis NL, Samuelson A (2007) The Insect and Spider Collections of the World website of the Bishop

Museum, Honolulu, Hawaii. http://hbs.bishopmuseum.org/codens/codens-r-us.html [accessed on Oct.

7, 2013]

42 RAJAEI & LASZLO: Scotopteryx kurmanjiana, a new species from the Kopet-Dagh Mountains

Parsons MS, Scoble MJ, Honey HR, Pitkin LM, Pitkin RB (1999) The Catalogue. In Scoble MJ (ed).

Geometrid Moths of the World: A catalogue (Lepidoptera, Geometridae). Collingwood, CSIRO Pub-

lishing, 1046 pp.

Rajaei Sh. H, Stiining D (2012) Scotopteryx kuznetzovi (Wardikian, 1957) (Lepidoptera, Geometridae,

Larentiinae), a new species for the fauna of Iran and Turkey. Bonn zoological Bulletin 61 (1): 135-139.

Rajaei Sh. H (2012) Modules to the Biodiversity of the Geometridae of Iran (Lepidoptera), using classical

methods and DNA techniques (Larentiinae and Ennominae partim). PhD thesis, Bonn university-Ger-

many.

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crolepidoptera. Entomologist’s Gazette 27: 127-132.

Schmidt O (2013) The structure and musculature of male terminalia in the tribe Xanthorhoini Pierce and

related tribes (Lepidoptera: Geometridae: Larentiinae), with particular reference to the Palaearctic and

Australian regions. Zootaxa 3721 (6): 552-572.

Scoble MJ, Hausmann A [updated 2007] Online list of valid and available names of the Geometridae

of the World, http://lepbarcoding.org/geometridae/species_checklists.php [website visited March 5,

2012]

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3136: 1-44.

Wardikian CA (1957) Novyj vid pjadenitsy iz roda Ortholitha Hb. (Lepidoptera, Geometridae) iz Arm-

janskoj SSR [A new geometrid moth species of the genus Ortholitha Hb. of the Armenian SSR].

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nischen SSR] 25: 281-283. [in Russian]

Nota Lepi. 37(1) 2014: 43-48 | DOI 10.3897/n1.37.7953

Description of the reduced mouth parts of Coleophora micronotella Toll

(Lepidoptera, Coleophoridae), with a new synonym

BERNARD LANDRY!, GIORGIO BALDIZZONE”

1 Muséum d'histoire naturelle, C.P. 6434, CH-1211 Geneva 6, Switzerland; bernard.landry@ville-ge.ch

2 via Manzoni 24, 14100 Asti, Italy; giorgiobaldizzone@tin.it * Contribution to Coleophoridae knowledge

no. CXXVII

http://zoobank.org/103A6A77-76A0-4416-AEEF-1DD57AC3A019

Received 13 November 2013; accepted 12 March 2014; published 15 June 2014

Subject Editor: Lauri Kaila

Abstract. The reduced mouth parts of Coleophora micronotella Toll (Coleophoridae) are described and

illustrated. The proboscis is lacking, but rudimentary maxillary and labial palpi are present. The mouth

parts of C. micronotella are compared with those of other species of Coleophora, including some others

with reduced mouth parts, and their possible adaptive value is discussed. Ramidomia Falkovitsh, 2005 and

Ramidiomia Falkovitsh, 2005, a misspelling, are synonymized with Coleophora Hiibner, 1822.

Résumé. La morphologie des piéces buccales trés réduites de Coleophora micronotella Toll (Coleopho-

ridae) est décrite et illustrée. Cette espèce présente des palpes maxillaires et labiaux rudimentaires, mais

aucun proboscis. Les pièces buccales de C. micronotella sont comparées avec celles d’autres espèces de

Coleophora, incluant quelques-unes ayant des piéces buccales réduites, et la possible valeur adaptative

de la réduction des piéces buccales est discutée. Ramidomia Falkovitsh, 2005 et Ramidiomia Falkovitsh,

2005, à l’orthographe fautive, sont mis en synonymie avec Coleophora Hiibner, 1822.

Introduction

In Glossatan Lepidoptera, which comprise over 99.9% of the species of the order, the mouth

parts typically consist of a labrum bearing a pair of lateral pilifers anteriorly, a haustellum (pro-

boscis) formed by the modified galeae of the maxillae and a pair of maxillary palpi medially,

and a pair of labial palpi posteriorly. Mandibles are fully formed only in the non-glossatan

Micropterigidae and Heterobathmiidae. In some scattered groups of species of various families

and in some larger groups, such as a good proportion of Lasiocampoidea and Bombycoidea,

however, a reduction of the mouth parts can be observed. This phenomenon has not been dealt

with comprehensively in the literature and we offer some observations in an attempt to explain

it in one small species.

As shown in detail by Capuse (1971), the members of Coleophoridae, although generally

quite small, usually have well-developed mouth parts, especially in the form of prominent,

recurved, tri-segmented labial palpi and a functional haustellum. Some species with reduced

mouth parts are nevertheless known, such as C. minipalpella Baldizzone, 1998, described from

Andalusia, Spain and known from only four specimens. Here we describe and illustrate the ves-

++ LANDRY & BALDIZZONE: Description of the reduced mouth parts of Coleophora micronotella Toll...

tigial mouth parts of Coleophora micronotella Toll, 1956, for which there are more specimens

available for study.

In the original description, Toll (1956) mentioned the colour of the head and antennae, and

indicated that the palps are missing in all of the nine specimens of C. micronotella that he had

at hand, but said nothing about the lack of proboscis. Falkovitsh (2005) erected the monotypic

genus Ramidomia, here synonymized with Coleophora Hübner, 1822 (along with Ramidiomia

Falkovitsh, 2005, a misspelling) to accommodate C. micronotella. Along with Baldizzone et

al. (2006) and others, supported by the phylogenetic analysis of Bauer et al. (2012), we find

it more coherent to use one large genus Coleophora (1340+ described species) than a classi-

fication of Coleophora subdivided in numerous small genera that have been erected without

comprehensive phylogenetic inference, as exemplified by Falkovitsh (2005). The latter author

noted that C. micronotella has short palpi, with the third segment shorter than the second, but

did not mention the lack of proboscis. Subsequent records of the species did not add any more

morphological information (Baldizzone 1994; Baldizzone et al. 2006).

Methods

The head of one specimen was mounted on slide in Euparal after maceration in about 15%

KOH at 60°C in an oven for an hour, cleaning in 30% ethanol, staining in Orange G and chlora-

zol black, and dehydrating in baths of 95% ethanol and isopropanol for one hour each.

The photographs of the scaled head as well as that of the slide-mounted whole head were

taken with a Leica MZ APO dissecting scope and AutoMontage, the close-ups of the mouth-

parts on slide with a Zeiss microscope and AutoMontage, and the scanning electron micro-

graphs with a Zeiss DSM 940-A scanning electron microscope in the Muséum d’histoire natur-

elle de Genève (MHNG).

Coleophora micronotella Toll, 1956

Figs 1-4

Material. 2 4 (one dissected, slide MHNG ENTO 5970), © [Italy] ‘SARDEGNA, Orien. | La Caletta | 2. VIII.81-

lux | leg.Baldizzone’; 1 4 (head prepared for SEM), same data except ’29.VII.81’; 2 4 (one dissected, slide MHNG

ENTO 7501), ‘SARDEGNA, Merid. | Porto Botte | 23.VII.81-lux | leg.Baldizzone’, deposited in MHNG. 1 ¢ (dis-

sected, slide MHNG ENTO 7503) [Tunisia] ‘Mauretania | Tunesia-Sud | Oase Tozeur | 28.4.-11.5.1981 | leg. M. u. W.

Glaser’, deposited in MHNG. Additional material, undissected: 194 specimens from Sardinia (leg. Baldizzone); 30

from Tunisia (24 from Tozeur, 1921, leg. Dumont; 7 from Tozeur e Gabes, 1980, leg. Glaser); deposited in collection

Baldizzone.

Description of head. Frontal aspect with appressed scales, with short, apparent scaled

palpi, without proboscis. Ventral edge of fronto-clypeus slightly sinuate. Labrum not ap-

parent. Maxillary palpi unsegmented, very short, with few scale sockets and setae, with

apical sensillum styloconicum. Labial palpi slightly longer than maxillary palpi, reaching

apices of subgenal apophyses, with two segments visible, apical segment about half as long

as basal segment and lacking vom Rath’s organ. With sac-like rounded structure laterally

from labial palpi.

Nota Lepi. 37(1) 2014: 43-48 45

Figures 1-4. Head of Coleophora micronotella Toll. 1. Frontal view. 2. Scanning electron micrograph

(fc: fronto-clypeus; Ip: labial palp, partly collapsed; mp; maxillary palpus; se: sensillum styloconicum). 3.

Ventral view showing labial palpi. 4. Dorsal view showing maxillary palpi.

Notes. The halophilous biotope of La Caletta, where three of the specimens of C. micronotel-

la studied were collected, is about one kilometer away from the Mediterranean, on the sides of

a small canal where tamarix (Tamarix sp., Tamaricaceae) grows. The specimens were collected

with a black light in a salt marsh where Salicornia (Amaranthaceae) and other halophilous her-

baceous plants were growing. Hundreds of specimens came to light, but could not come to rest

on the sheet because of the presence of thousands of staphylinid beetles and flies. Falkovitsh

(2005) recorded that the larva of C. micronotella feeds as a borer, without making a case, in

‘green branches (assimilating shoots)’ of Halostachys caspica C.A. Mey. and Halocnemum

strobilaceum (Pallas) Bieb. (Amaranthaceae).

Described from Tozeur, Tunisia, the species is also found in Italy (mainland, Sardinia, Sici-

ly) and Spain. It has also been reported from Iran (Baldizzone 1994), and Algeria, Kazakhstan,

and Tadjikistan (Falkovitsh 2005). However, a specimen from the Iranian series mentioned in

Baldizzone (1994) was re-examined and found to represent another, probably undescribed spe-

cies. Its mouth parts are more developed than in C. micronotella, with the labial palpi about as

long as 3/4 the widest diameter of the compound eye, but without apparent proboscis. Also, the

male valva differs slightly in the distally larger and more evidently separated sacculus from the

basally narrower valvula, and the phallus is narrow, needle-like, instead of spatulate.

46 LANDRY & BALDIZZONE: Description of the reduced mouth parts of Coleophora micronotella Toll...

Other species of Coleophora with vestigial mouth parts

Coleophora galligena Falkovitsh, 1970, described from Uzbekistan, has vestigial palpi and

no proboscis. Coleophora daeva Baldizzone, 1994, C. minipalpella Baldizzone, 1998, and C.

zagella Falkovitsh, 1972, respectively described from Iran, Spain, and Mongolia, have short

proboscis and palpi. These four species and C. micronotella can be associated with the 8" group

of Toll (1953, 1962), based on their simple genitalia, but Toll (1956) only mentioned that C.

micronotella is close to C. plurifoliella Chrétien, and never associated this species with any

group subsequently. These species are associated with halophilous habitats and deserts and they

were not treated by Bauer et al. (2012).

Discussion

The sac-like rounded structures that protrude laterally from the labial palpi in C. micronotella

may represent vestigial mandibles. Rudimentary mandibles were found in all species of Co-

leophora studied by Capuse (1971), with variation in shape and size. Capuse (1971) noted that

the mandibles are usually sclerotized, but he found at least one species in which they were

membranous (C. asthenella Constant). Capuse (1971) did not study C. micronotella, but for

comparison it may be noted that among the 100 species studied he mentioned that the maxillary

palpi were missing in one only (C. asthenella). Although it is often difficult to count the number

of maxillary palp segments precisely because the limits between segments may be not easily

discernible, Capuse found that the number of segments varied from one to five, the one-seg-

mented species numbering 20, the 2-segmented 31, and the other cases in lower frequencies.

Capuse (1971) also found that C. ciconiella Zeller, a species with two maxillary palpomeres,

has the apex of the maxillary palpi adorned with a sensillum styloconicum.

The labial palpi in the Lepidoptera groundplan are tri-segmented (Kristensen 2003). This

is the condition found in Coleophora, but the overall length of the palpi, length of individual

palpomeres, shape of terminal palpomere, and vestiture vary across species. Capuse (1971)

did not study any species with markedly reduced labial palpi. The lack of vom Rath’s organ is

unusual in Coleophoridae and presumed here to be linked with the reduction of the labial palpi.

Regarding the proboscis, Capuse (1971) recorded no species in which it is lacking, but a few

in which it is slightly shorter than the labial palpi, and membranous. No species with reduced

mouth parts were found from the Western Hemisphere (J.-F. Landry, pers. comm. to G. Baldiz-

zone), but several undescribed species from the Afrotropical region have reduced proboscis

and palps.

Bauer et al. (2012) presented the first phylogeny of Coleophoridae based on a formal analy-

sis of molecular characters; one hundred and two European species were analysed, but neither

C. micronotella, any of the above-mentioned species with reduced mouth parts, nor any species

of Toll’s 8" group were included. Nevertheless, two of the species groups recovered, the albel-

la and alticolella groups, have species feeding on Amaranthaceae, with the larvae of the a/bella

group being internal miners in the first larval stages, until they become too large to be fully hid-

den in the nutritive tissue, after which they build a tubular silk case. This host plant choice and

the internal feeding habits may point to an affinity of C. micronotella with the albella group.

Nota Lepi. 37(1) 2014: 43-48 47

Although the reduction or loss of the proboscis occurs in many glossatan lineages (Kris-

tensen 2003: 54), the adaptations related to the absence of a functional proboscis are unclear,

and no discussion of this phenomenon was found in the literature. Cäpuse (1971: 23) believed

that in species with reduced galeae, hence without a functional proboscis, the reduction is cor-

related with the digestive tract, without qualifying this correlation.

Several species of Saturniidae have reduced mouth parts and do not feed as adults (Taylor

1957), presumably to place most of the energy resources acquired during the development of

the larva into the production of a larger quantity of eggs and/or into the development of a more

efficient mate locating mechanism, hence the highly plumose male antennae. A shift in energy

resource allocation may be the reason for reduced mouth parts in Coleophora micronotella and

others, but it may also be that these species live in habitats where food sources (water, nectar)

for adult Lepidoptera with low vagility are too scarce or poisonous, such as with a deleterious

salt concentration, for example. The fact that C. micronotella and other species with reduced

mouth parts live in halophilous or desert biotopes, and that the larva of C. micronotella is a

borer (a strategy to avoid ingesting too much salt and/or to secure enough water, perhaps) lend

support to this hypothesis. We believe that this phenomenon is akin to the loss of functional

wings in some Lepidoptera species living on small, wind-swept islands (e.g., Davis & Mendel

2013) or the loss of functional eyes in cave-dwelling animals.

Acknowledgements

Yuriy Budashkin, Ole Karsholt, Niels Peder Kristensen, Jean-François Landry, Rodolphe Rougerie, Jukka

Tabell, and Hugo van der Wolf replied to our requests for information. Florence Marteau and André Piuz

of MHNG respectively assembled the plate and did the scanning electron micrographs. Jean-François

Landry and two anonymous referees provided useful comments on the manuscript. We thank them all

very much for their work in our favour.

References

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Irano-Anatolica e regioni limitrofe (Lepidoptera). Associazone Naturalistica Piemontese, Memorie vol

3. Apollo Books, Stenstrup. 424 pp.

Baldizzone G, van der Wolf H, Landry J-F (2006) Coleophoridae, Coleophorinae (Lepidoptera). In:

Landry B (Ed) World Catalogue of Insects, Vol. 8. Apollo Books, Stenstrup. 215 pp.

Bauer F, Stiibner A, Neinhuis C, Nuss M (2012) Molecular phylogeny, larval case architecture, host—

plant associations and classification of European Coleophoridae (Lepidoptera). Zoologica Scripta 41:

248-265. doi: 10.1111/j.1463-6409.2012.00532.x

Capuse I (1971) Recherches morphologiques et systématiques sur la famille des Coleophoridae

(Lepidoptera). Bucarest. 116 pp.

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ery of a new brachypterous species (Lepidoptera, Tineidae). ZooKeys 341: 1-20. doi: 10.3897/zook-

eys.341.6146

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49: 869-885. [In Russian]

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Palaearctic. Entomologicheskoe Obozrenie 84: 167—176. [In Russian]

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Kristensen NP (2003) 4. Skeleton and muscles: adults, pp. 39-131. In: Kristensen NP (Ed) Handbook of

Zoology, Vol. IV, Part 36, Lepidoptera, moth and butterflies, Vol. 2, Morphology, physiology, and

development. W. de Gruyter, Berlin, New York.

Taylor JS (1957) Notes on the proboscis in the Saturniidae. Entomologist’s Record and Journal of Vari-

ation 69: 151-153.

Toll S (1953) Rodzina Eupistidae polski. Documenta Physiographica Poloniae 32 [1952]: 293 pp. + 38

pls.

Toll S (1956) Etude de quelques Coleophoridae d’ A frique du Nord et leurs genitalia (Lep). L’ Entomologiste

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Nota Lepi. 37(1) 2014: 49-62 | DOI 10.3897/n1.37.7929

On the taxonomic status of Ochromolopis ictella (Hübner, 1813)

and ©. zagulajevi Budashkin & Sachkov, 1991

(Lepidoptera, Epermeniidae)

REINHARD GAEDIKE!, RICHARD MALLY?

1 Florusstraße 5, 53225 Bonn, Germany, tinagma@msn.com

2 University Museum of Bergen, Natural History Collections, Allegaten 41, 5007 Bergen, Norway;

richard.mally@um.uib.no

http://zoobank. org/D5 18C8D6-3E65-437E-AA4A-E9212D34C085

Received 24 September 2014; accepted 17 March 2014; published: 15 June 2014

Subject Editor: Vazrick Nazari

Abstract. A detailed study of specimens from several regions of the distribution of Ochromolopis zagu-

lajevi Budashkin & Sachkov, 1991 and O. ictella (Hübner, 1813) shows that O. ictella and O. zagulajevi

are parapatric species with overlapping distribution in the Balkan Peninsula. Details of morphological and

molecular differences as well as a distribution map with locations of the examined specimens are given.

Introduction

Ochromolopis Hübner, 1825 is one of the 11 described genera of Epermeniidae Spuler, 1910,

a family currently comprising 188 species and distributed worldwide. It is the only family

within the superfamily Epermenioidea Minet, 1983, which, according to Dugdale et al. (1998),

shows indications of affinities with the alucitoid/pterophoroid assemblage, but the family

placement has not received any support in recent molecular studies across all of Lepidoptera

(Mutanen et al. 2010; Regier et al. 2013). The potential autapomorphies of Epermeniidae are

the following: hind tibia with stiff bristles; forewing fringe with groups of lamellar scales; lar-

val submentum with posterior protuberance; prothoracic prespiracular L group bisetose; pupa

unspined, abdominal segments I-IV immovable, segment IX with characteristic paired lateral

pits (Dugdale et al. 1998). The majority of species bear tufts or raised scales on the dorsum of

forewings. Further characteristics are the loss of one of the apical forewing veins (M3 and Cul

fused) and the widening of the ventral branch of anterior apophysae in females. The genus

Ochromolopis currently comprises 11 species, with four of them distributed in the Palaearctic

region, while the others have a Nearctic, Afrotropical and Oriental distribution.

We examined Ochromolopis ictella (Hiibner, 1813) and O. zagulajevi Budashkin &

Sachkov, 1991. The two species are closely related and not distinguishable superficially.

Only the genital morphology shows clear differences. A more detailed study was made to

determine the variability within the two taxa not only by using the traditional methods of

morphological investigation but also by means of molecular methods (DNA Barcoding) by

the second author.

50 GAEDIKE & MALLY : On the taxonomic status of Ochromolopis ictella (Hübner, 1813)...

Material and methods

The examined material originates from numerous collections and was provided by museum

curators as well as by private collectors. A list of examined material is given in the appendix.

Morphological methods

Genitalia of both sexes were dissected in order to study morphological variability. Phallus

and valvae were removed from the genitalia capsule (uncus-tegumen-vinculum with saccus)

during dissection. The ring-shaped connection of tegumen-vinculum was not cut laterally but

kept intact. Drawings (all at the same scale) were made from genitalia of the two taxa and their

variation was compared.

Molecular methods

For the molecular investigation of relationships between Ochromolopis ictella and O. zagu-

lajevi we analysed the Barcode fragment of the mitochondrial COI gene. In order to obtain

a high quantity of DNA, we performed the DNA extraction on the abdomen of dried speci-

mens, followed by genital dissection from the macerated abdomen, as suggested by Knölke

et al. (2005). DNA was extracted using the Macherey-Nagel NucleoSpin Tissue kit according

to the manufacturer’s suggestions. PCR amplification of an approximately 630 base pair

(bp) fragment of the COI barcode sequence was done with the primer pair HybLCO/Nancy

or in the case of fragmented DNA with the primer pairs HybLCO/K699 and Ron/Nancy,

respectively (Folmer et al. 1994; Mitchell et al. 2005; Simon et al. 1994). The sequences

were amplified with BIO-X-ACT Short DNA polymerase (Bioline). The PCR program for

BIO-X-ACT Short DNA polymerase corresponds to the manufacturer’s guidelines in con-

junction with the respective primer annealing temperature of 48°C. The PCR products were

checked for amplification success via gel electrophoresis on a 1% agarose gel, subsequent

staining with GelRed, and final examination under UV light. The PCR products were cleaned

with ExoSAP-IT (USB Corporation). The sequence PCR was performed with BigDye Ter-

minator v3.1 Cycle Sequencing Kit (Applied Biosystems). After final sodium acetate-ethanol

clean-up of the samples, sequencing was carried out on a 3130 Genetic Analyzer (Applied

Biosystems). A Mastercycler ep gradient S (Eppendorf) was used for PCR amplifications,

ExoSAP-IT clean-up and sequence PCR.

Sequence alignment was carried out manually with PhyDE 0.9971 (Miiller et al. 2008). Cal-

culation of the genetic distances as well as of the dendrogram using the Neighbor-Joining (NJ)

method (Saitou & Nei 1987; Studier & Keppler 1988) were performed under the uncorrected-p

(uncorr-p) model (Srivathsan & Meier 2012) in PAUP* 4.0b10 (Swofford 2002). Ochromol-

opis kaszabi Gaedike, 1973 was included in the analysis as an outgroup and for comparison of

the genetic distances of the two investigated taxa to a more distantly related species.

Nota Lepi. 37(1) 2014: 49-62 51

Distribution mapping

In order to examine the distributional pattern of Ochromolopis ictella and O. zagulajevi,

collection localities were compiled from labels of studied specimens and from literature.

Geographical coordinates of these collection localities were obtained via Google Earth, Ver-

sion 5.2.1.1588 and subsequently plotted on a map using DIVA-GIS, Version 7.2.3 (Hijmans

et al. 2004).

Abbreviations

coll. Arenberger Ernst Arenberger, Vienna, Austria

coll. Bengtsson Bengt A. Bengtsson, Farjestaden, Sweden

coll. Schmitz Willibald Schmitz, Bergisch-Gladbach, Germany

ETHZ Eidgenössische Technische Hochschule, Zürich, Switzerland

FMNH Finnish Museum of Natural History, Helsinki, Finland

HNHM Hungarian National History Museum, Budapest, Hungary

LMAD Löbbecke Museum und Aquazoo, Düsseldorf, Germany

MNG Museum der Natur, Gotha, Germany

MTD Museum fiir Tierkunde, Senckenberg Naturhistorische Sammlungen Dresden, Germany

NHMB Naturhistorisches Museum Basel, Switzerland

NHRS Naturhistoriska Riksmuseet, Stockholm, Sweden

NMEG Naturkundemuseum, Erfurt, Germany

NMPC National Museum (Natural History), Prague, Czech Republic

NMW Naturhistorisches Museum, Vienna, Austria

SDEI Senckenberg Deutsches Entomologisches Institut, Müncheberg, Germany

SMNK Staatliches Museum fiir Naturkunde, Karlsruhe, Germany

SMNS Staatliches Museum fiir Naturkunde, Stuttgart, Germany

TLMF Tiroler Landesmuseum Ferdinandeum, Innsbruck, Austria

ZIN Zoological Institute, Russian Academy of Sciences, St. Petersburg, Russia

ZMHB Museum fiir Naturkunde der Humboldt-Universitat, Berlin, Germany

ZMUC Zoological Museum, Copenhagen, Denmark

ZSM Zoologische Staatssammlung, Munich, Germany

Results

Redescription of morphology

The two taxa do not differ in superficial appearance (see Fig. 1 for a specimen of O. ictella).

Head, thorax, abdomen dark grey, shiny, forewing lead-grey, shiny, with two longitudinal gold-

en-yellowish stripes: the first obliquely extending from base to dorsum shortly before half of

wing, where first tuft of raised scales is situated, the second running parallel, initiating at 1/3 of

cell, pointing straight towards outer margin and ending well before apex. The distal end of first

stripe connected with the proximal end of the second stripe, forming a sustained Z. O. ictella and

O. zagulajevi only differ in the shape of the genitalia (ictella: Figs 2-9, 19-21, 30; zagulajevi:

Figs 10-18, 22-29, 31-32).

According to Budashkin & Sachkov (1991), O. zagulajevi has an area of numerous strongly

sclerotized cornuti in the phallus within the posterior half of the vesica (Fig. 12). We found that,

92 GAEDIKE & MALLY : On the taxonomic status of Ochromolopis ictella (Hübner, 1813)...

Figure 1. Ochromolopis ictella, imago.

Figures 2-9. Male genitalia of Ochromolopis ictella. 2-4. specimen from Zljeb, Montenegro: 2. un-

cus-tegumen-vinculum. 3. valva. 4. phallus. 5-9. variability in socii shape: 5. Xauen A’Faska, Maureta-

nıa. 6. Piedmont, Italy. 7. Neustadt, Germany. 8. Naumburg, Germany. 9. Vienna, Austria.

Nota Lepi. 37(1) 2014: 49-62 53

Figures 10-18. Male genitalia of Ochromolopis zagulajevi. 10-12. specimen from Crimea, Ukraine: 10. un-

cus-tegumen-vinculum. 11. valva. 12. phallus. 13-18. variability in socii shape: 13. Danubian delta, Romania.

14-15, 17. Kabardino-Balkarskij Nat. Res., Russia. 16. Crimea, Ukraine. 18. Djanik, Turkey.

due to the posteriad evertability of the vesica, the position of the cornuti may vary between differ-

ent genital dissections. Characteristic for zagulajevi is the large quantity of cornuti.

In most cases the cornuti are in a more or less compact cluster, concentrated in the proximal

fourth of the phallus, but in some cases the area of cornuti in the vesica covers the second- to

54 GAEDIKE & MALLY : On the taxonomic status of Ochromolopis ictella (Hübner, 1813)...

Figures 19-29. Variability in costa shape. 19-21. variability within Ochromolopis ictella: 19. Neustadt,

Germany. 20. Vienna, Austria. 21. Naumburg, Germany. 22-29. variability within O. zagulajevi: 22-24.

Kabardino-Balkarskij nat.res., Russia. 25. Northern Adriatic, Croatia. 26. Crimea, Ukraine. 27. Djanik,

Turkey. 28. Gjalica Ljums, Albania. 29. Danubian delta, Romania.

third-fourth of the phallus length. The vesica of ictella also exhibits minute cornuti, but their

number is mostly smaller, and they are not arranged as compactly as in zagulajevi (Fig. 4).

The shape of the narrow socii allows for an easier differentiation. In zagulajevi the socii

have a dorsally attached lobe-shaped process at half of their length, which varies in size and

shape (Figs 13-18). In contrast, in ictella this process is thorn- or hook-shaped and has a point-

ed tip, and is also variable in size and shape (Figs 5—9). The shape of the costal arm of the valva,

which is nearly as long as the valva itself, is variable in both taxa. However, the costal arm

appears to be more compact in ictella than in zagulajevi.

The female genitalia of the two taxa also exhibit some structural differences: in O. zagulaje-

vi, the posterior part of ductus bursae is more strongly sclerotized, but sometimes the posterior

sclerotization of the ductus is not developed (compare Figs 31 and 32); the median part is

wrinkled and covered with numerous minute semicircular sclerotizations. In contrast, O. ictella

lacks strong sclerotization in the posterior ductus bursae and the wrinkles at the median part are

weakly developed and have minute sclerotizations (Fig. 30).

In order to evaluate the significance of the investigated morphological characters for de-

fining the taxonomic status of the two taxa it was deemed important to include a third taxon

into the treatment, Ochromolopis kaszabi Gaedike, 1973. This species was described from

Mongolia, and its currently known distribution ranges from Altai through Mongolia to Russian

Far East and China. Superficially, O. kaszabi is not distinguishable from the above mentioned

taxa, but it shows clear differences in the male genital structures: socii are broad, more or less

parallel, nearly obliquely truncated, with a more or less pointed tip, depending on preparation

Nota Lepi. 37(1) 2014: 49-62 55

Figures 30-32. Female genitalia. 30. Ochromolopis ictella (Abruzzi, Italy). 31-32. variability within O.

zagulajevi: 31. Crimea, Ukraine. 32. Greece.

—

Figures 33-38. Male genitalia of Ochromolopis kaszabi (Mongolia; according to Gaedike, 1973). 33.

uncus-tegumen-vinculum. 34. valva. 35. phallus. 36. variable form of socius. 37-38. variability of valva

(Hoengshan, China).

56 GAEDIKE & MALLy : On the taxonomic status of Ochromolopis ictella (Hübner, 1813)...

Table 1. DNA barcoding specimen information.

Taxon Dis span] Origin, date, collector Sequence Sanlzanls

voucher length | accession no.

MTD Lep1073 Italy, Piedmont, Valdieri, reserve, 850m 29.-30.v1.2008, leg. 612 bp KF511936

O. Karsholt

ZSM Lep 27010 Germany, Bavaria, Bar 400m, 08.vi.1994, 658 bp | HM902062

ictella = ==

Macedonia, Mavrovo NP, Korab, summit ridge, 2700m,

TLE Len 0228 28.vii.2011, leg. P. Huemer & G. Tarmann 658 bp | Shonen)

Macedonia, Mavrovo NP, Korab, summit ridge, 2700m,

NER 28.vi1.2011, leg. P. Huemer & G. Tarmann a re

MTD Lep1071 Croatia, Istria, Belavici, Sn 08.-14.1x.2008, leg. W. 612 bp KF511934

MTD Lep1072 Croatia, Istria, Belavici, ae 08.-14.1x.2008, leg. W. 612 bp | KF511935

BEER. \ 1 ee

MTD Lep1074 Italy, Lucania, Mt. Pollino, 780m, 03.x.2010, leg. P. Skou 612 bp KF511937

MTD Lep1075 Italy, Lucania, Mt. Pollino, 780m, 03.x.2010, leg. P. Skou 612 bp KF511938

MTD Lep1076 SW Bulgaria, Pirin Sandanski, Ilindentsi, 500m, 28.11.04. 612 bp KFS11939

a SE BI Pin Gander Vincent, A REDE

ulgarıa, Pırın Sandanski, Ilindentsi, m, 26.111. .

MTD Lep1077 iv2011. les! NY Savenkoy 612 bp KF511940

MTD Lep1078 SW Bulgaria, Pirin, Sandanski, Ploski, 250m, 17.-31.v.2010, 612 bp KFS11941

leg. N. Savenkov

SW Bulgaria, Pirin Sandanski, Ilindentsi, 500m, 28.11.04.

MTD Lep1079 iv 2011, leg N Savenkov 612 bp KF511942

MTD Lep1080 SW Bulgaria, Pirin, Sandanski, Ploski, 250m, 17.-31.v.2010, 591 bp KFS11943

leg. N. Savenkov

aaa MTD Lep1081 Russia, Siberia, Chita, Bun a: 27.v11.1997, leg. I. 612bp | KF511933

(see Figs 33 and 36). The costal arm of the valva is about two thirds of the valval length and

has a forked distal end with a short ventral branch and a longer costal branch with pointed tip,

directed more or less obliquely upwards, edges of the two branches with minute pointed thorns.

The shape of the costal arm is variable (see Figs 37-38). Female genitalia (according to the

description in Budashkin & Sachkov 1991: figs 3, 4) are characterised by the ductus bursae

lacking strong sclerotization, but with strongly sclerotized sternal segments.

Molecular results

The barcode sequences length was 591-658 bp (see Table 1). No indels have been observed in

any of the sequences. All obtained barcode sequences have been submitted to GenBank (for

accession numbers see Table 1).

In the NJ analysis we obtained two clusters comprising 6 and 7 samples, respectively (Fig.

39). The two clusters do not correspond to the two taxa. Rather, in one cluster there are two

Ochromolopis ictella and five O. zagulajevi specimens, and in the other cluster there are two

O. ictella and four O. zagulajevi specimens. In O. zagulajevi we observed two cases in which

specimens from the same locality and period of collection are not found in the same Barcode

cluster: 1) Lep1075 versus Lep1074 (Italy, Mt. Pollino) and 2) Lep1076 & Lep1079 versus

Lep1077 (Bulgaria, Ilindentsi).

Intraspecific divergences within ictella range from 0.168% to 1.672% and in zagulajevi from

0% to 1.359% (see Table 2). Interspecific uncorr-p divergences between ictella and zagulajevi

Nota Lepi. 37(1) 2014: 49-62 37,

kaszabi_1081_Russia_Chita

ictella_BC_ZSM_Lep_27010_Germany_Nittendorf

ictella_1073_Italy_Valdieri

zagulajevi_1078_Bulgaria_llindentsi

zagulajevi_1079_Bulgaria_llindentsi

zagulajevi_1080_Bulgaria_llindentsi

zagulajevi_1076_Bulgaria_llindentsi

zagulajevi_1075_Italy_MtPollino

ictella TLMF Lep 05228 Macedonia_Korab

ictella TLMF Lep 05229 Macedonia _Korab

zagulajevi 1074 Italy _MtPollino

zagulajevi 1077 _Bulgaria_llindentsi

zagulajevi_ 1071 _ Croatia Marcana

zagulajevi_ 1072 Croatia Marcana

0.2

Figure 39. NJ dendrogram, based on uncorrected-p distances; scale bar represents 2.0% uncorrected-p

distance.

range from 0% to 1.528%. In contrast, O. kaszabi has a Barcode divergence of 2.922-3.100%

with ictella and 2.451-3.045% with zagulajevi.

Distribution map

The map of the investigated taxa (Fig. 40) shows that Budashkin & Sachkov’s (1991) presump-

tion in the original description of ©. zagulajevi concerning the distribution was right. All ex-

amined specimens from Caucasus region, Crimea, Iran, Turkey, and Southern Europe (Greece,

Bulgaria, Albania, S-Italy) belong to O. zagulajevi. On the contrary, all examined specimens

from North Africa, southwestern, central and northern Europe (Spain, France, Switzerland,

Denmark, Finland, Germany, Austria, N-Italy, Poland, Czech Republic, Slovakia, Hungary),

and Montenegro (only one specimen studied) belong to O. ictella.

58 GAEDIKE & MALLY : On the taxonomic status of Ochromolopis ictella (Hübner, 1813)...

Table 2. Uncorrected-p sequence divergence matrix with divergence values as percentage.

1081 1073 127010 105228 |05229 |1071 11072 |1074 11075 11076 |1077 |1078 |1079

ictella MTD

Lep1073 za |

ictella ZSM

27010 3.083 10.168

ictella TLMF

05228 3.100 1.476 |1.672 ee

ictella TLMF 2 922 1.299 |1.520 10.760

05229

zagulajeviMTD |) 451 11.144 [1.295 |0.654 |0.477

Lep1071

zagulajevi MTD

Lep 1072 2.451 |1.144 |1.295 0.000

zagulajevi MTD

Lep1074 2.941 |1.307 |1.465 0.490 |0.490

zagulajeviMID | o41 ]0.327 |0.490 1.144 |1.144 |1.307

Lep1075

zagulajeviMID |) 758 10.163 [0.326 0.980 |0.980 |1.144

Lep1076

zagulajevi MID Dogs |1.307 |1465 0.490 |0.490 |0.327

Lep1077

zagulajeviMID |, 941 10.000 |0.168 1.144 11.144 |1.307

Lep1078

zagulajeviMTD | 941 0.000 |0.168 1.144 |1.144 |1.307

Lep1079

zagulajevi MTD

a 3.045 |0.000 [0.172 1.187 |1.187 [1.359 |0.338 0.170 |1.356 |0.000 [0.000

In Croatia and in Macedonia both taxa occur sympatrically. Additional sympatrical distribu-

tion might be present in Italy, where O. ictella is present from the northern part southwards to

Umbria, and O. zagulajevi in the southern regions and Sicily. No material was available from

Slovenia.

Discussion

The comparison of genital morphology between the two species reveals broad concordance

of the investigated structures. Only one differing feature was found between O. ictella and

O. zagulajevi, namely the shape of the socii within male genitalia. The divergence in genital

morphology is evidently larger between O. kaszabi and the O. ictella-zagulajevi complex than

between ictella and zagulajevi.

The analysis of DNA Barcodes reveals that the range of interspecific Barcode divergence

between ictella and zagulajevi (0-1.528%) is within the range of intraspecific divergence of

0.168-1.672% in ictella and 0-1.359% in zagulajevi. This suggests that it is possible that these

two taxa might actually represent one somewhat variable species. However, the finding of one

constant morphological difference in the male genitalia between ictella and zagulajevi and the

sympatric occurrence of both taxa on the Balkan peninsula imply the validity of their species

Status.

Nota Lepi. 37(1) 2014: 49-62 39

Figure 40. Distribution map of Ochromolopis ictella (red circles) and ©. zagulajevi (black triangles) in

the West Palaearctic.

Until further molecular work with much greater specimen sampling, focusing on estab-

lishing reasons behind the two DNA barcoding clusters (e.g., incomplete lineage sorting;

Funk & Omland 2003), can provide more data on the status of the two species, we recom-

mend no change in their taxonomic status. Future work should also aim to infer a more accu-

rate distribution border between these two taxa in Europe, especially in the areas of Central

Italy southwards, of the Balkan Peninsula and from Slovenia and Hungary eastwards.

Acknowledgements

The study was only possible through the kind support of numerous entomologists by loaning material

of the examined taxa. For this important help we thank the custodians of the museums listed in the

Abbreviations section as well as Ernst Arenberger (Vienna, Austria), Bengt A. Bengtsson (Färjestaden,

Sweden), Hartmut Roweck (Kiel, Germany) and Willibald Schmitz (Bergisch-Gladbach, Germany). For

the provision of DNA Barcoding data and the respective specimens for their use in this study we thank

Peter Huemer (Innsbruck, Austria) and Andreas Segerer (Munich, Germany). We thank Matthias Nuss

for the support of the molecular analyses in the DNA laboratory of the MTD and Christian Kutzscher

(SDEI Miincheberg) for making the colour picture. We are also thankful to the reviewers for their valuable

comments.

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Nota Lepi. 37(1) 2014: 49-62 61

Appendix

A list of examined specimens of Ochromolopis ictella (Hübner 1813), O. zagulajevi Budashkin

& Sachkov, 1991 and O. kaszabi Gaedike, 1973 follows below. All specimens were determined

by R. Gaedike or were part of the original description of zagulajevi. The list contains material

examined from approximately 1965 to 2013. The localities in this list are recorded as they

appear on the labels. The current depository of the material (if available) is provided using the

abbreviations listed in the Abbreviations section.

Ochromolopis ictella (Hiibner, 1813)

Mauretania, Xauen, A’Faska: 14, 20.vi.1931, leg. Reisser (NMW). Spain, Granada: 14, 19, 12.iv.,

22.vi., leg. Staudinger (ZMHB); Escorial: 14, vii.1924; Castellon, 3km S of Forcall: 1 9, 16.vi.1989,

leg. et coll. Bengtsson; Guadalajara, 1km NW Trillo: 14, 21.vii.1988, leg. Fibiger (ZMUC); Cuenca,

5km SW Huelamo, by Rio Jucar: 19, 19.vii.1988, leg. Fibiger (ZMUC); Sieddar Nevada, Camino de

la Veleta: 14, 19, 29.vii.1985, 3.vii.1986, leg. Traugott-Olsen (ZMUC). France, Bourgogne: 16, leg.

Constant (ZSM); Mt. Panaglia, env. of Nizza: 14, 3.x.1964, leg. Glaser (SMNK); Aude, Villedeigne: 14,

9.vu.1961, leg. Burmann (SMNK); La Voulte-sur-Rhône: 2 specimens, leg. Dresney; env. of Digne: Les

Mees: 14, 20.v.1977, leg. Bruer (ZSM); Basses Alpes, la Baume: 2 specimens, 26.vii.1973, leg. Groß

(LMAD); Alpes mar., St. Bres: 1 specimen, 24.vii.1973, leg. Groß (LMAD); Provence, 4km N Eyquians:

13, 4.vii.1989, leg. et coll. Bengtsson; Corsica (Rungs, 1988). Italy, Piedmont, Valdieri: 14, 29.-30.

vi.2008, leg. Karsholt (ZMUC); Abruzzi, Mte Sirente: 19, leg. Dannehl (ZSM); Liguria, Noli (Savona):

14,21.-30.ix.1951, leg. Klimesch (ZSM); Liguria, Andora: 19, 6.ix.1965, leg. Klimesch (ZSM). Mon-

tenegro, Zljeb, Neumontenegro: 14, 1916, leg. Penther (NMW). Macedonia, NP Mavrovo, Korab, sum-

mit ridge, ca. 2700-2750m, 20°32’48”E, 41°47’20”N: 44, 28.vii.—1.viii.2011, leg. Huemer & Tarmann

(TLMF). Austria, Wien: 4, leg. Mann (NMW); Lobau: 14, 19, viii.1916, leg. Predota (NMW); Möd-

ling: 14 (NMW); Hochzire/Tirol: 34, vii.1927, viii.1927 (NMW); Umgebung Seefeld: 14, 1.vii.1922,

leg. Bauer (ZSM); Innsbruck: 14, 14.vi.1938, leg. Burmann (SMNK); Lechtaler Alpen, 1700m: 14, 19,

5.—11.viii.1940, leg. Osthelder (ZSM); Pasterz: 14; Heiligenblut: 34, vii.1896; Dürnstein: 29, iv., vii.,

leg. Klimesch (ZSM); Brennersee, 1400m: 14, 14.vii.1968, leg. Burmann (SMNK); Stuben/Vorarlberg,

1500m: 1 specimen, 11.vi1.1962, leg. Groß (LMAD); Schütt near Villach: 1 specimen, 22.vii.1972, leg.

Groß (LMAD); Tirol, Tessenberg: 14, 12.-15.vii.1981, leg. Schnack (ZMUC). Switzerland, Kanton

St. Gallen Vättnerberg: 14, 12.viii.1909, leg. Müller-Rutz (NHMB); Vättis: 14, 19, vii., leg. Müller-

Rutz (NHMB); Kanton Graubünden, Endagin: Val Fuorn: 14, 19.vi.1905 (ETHZ); Ekschis, Safien: 1,

27.vi.1929, leg. Müller-Rutz (NHMB); Parpan: 14, 19, vi.1920, vii.1920, leg. Müller-Rutz (NHMB); Fe-

tan: 1 specimen, 31.vii. 1923, leg. Miiller-Rutz (NHMB); Remüs: 14,19, vii. 1933, viii. 1935, leg. Weber

(ETHZ); Mathon: 26, 3., 5.viii.1929, leg. Weber (ETHZ); Salorino: 14, 19, 26.vii.1926, 13.vii.1927,

leg. Weber (ETHZ). Czech Republic, Env. of Litomerice, Libochovany: 1 specimen, leg. Zimmermann

(NMPC); Zalezly: 1 specimen, leg. Wihan; Lednice: 1 specimen, leg. Zimmermann (NMPC); Hraba-

sice: 14, v.1977 (ZMUC). Slovakia, Zadiel: 1 specimen, leg. Povölny; Cenko: 1 specimen, leg. Patocka;

Banska Stiavnica: 1 specimen, leg. Patocka. Hungary, Budapest: 14, 18.v.1913, leg. Uhrik (HNHM);

Csakvar: 14, 7.vii.1961, leg. Gozmäny (HNHM); Puszta Peszer: 14, 10.vii.1929, leg. Osthelder (ZSM));

Bagloyirtas, Matra: 14, 12.vi.1951, leg. Gozmäny (HNHM). Germany, Halle/Saale: 1 specimen, leg.

Eichler (ZSM); Naumburg: 1 specimen, leg. Bauer (ZSM); Kyffhäuser: 5 specimens, leg. Hockermeyer /

Lenthe / Beer / Petry / Soffner (NMEG; SDEI); Ochsenburg/Kyffhäuser: 1 specimen, leg. Sutter (SMNK);

Bad Blankenburg: 1 specimen, leg. Steuer (ZMHB); Jena: 1 specimen, leg. Nikolaus (MNG); Inselsberg,

Georgenthal: 1 specimen, leg. Lenthe (MNG); Alter Stolberg near Nordhausen: 1 specimen, leg. Petry

(NMEG); env. of Erfurt: 1 specimen, leg. Beer (MNG); Löberschütz: 1 specimen, leg. Faulwetter; Tau-

tenburg: 1 specimen, leg. Faulwetter; Flachsleite: 1 specimen, leg. Faulwetter; Gleisberg: 1 specimen,

leg. Faulwetter; Totentäler, Kreis Nebra: 1 specimen, leg. Eichler (ZSM); Braunschweig: 1 specimen, leg.

62 GAEDIKE & MALLY : On the taxonomic status of Ochromolopis ictella (Hübner, 1813)...

Heinemann; Neustadt/Haardt: 1 specimen, leg. Eppelsheim (ZSM); Kaiserlautern: 1 specimen, leg. Heu-

ser; Grünstadt: 1 specimen, leg. Wörz (SMNS); Hambach: 1 specimen, leg. Wörz (SMNS); Battenberg:

1 specimen, leg. Wörz (SMNS); Wiesbaden: 1 specimen, leg. Wörz (SMNS); Stuttart: 1 specimen, leg.

Wörz (SMNS); Schelklingen: 1 specimen, leg. Wörz (SMNS); Baden: 1 specimen, leg. Hering (ZMHB);

Ascholding: 1 specimen, leg. Osthelder (ZSM); Steinebach: 1 specimen, leg. Osthelder (ZSM); Garchin-

ger Heide: 1 specimen, leg. Osthelder (ZSM); Garmisch: 1 specimen, leg. Osthelder (ZSM); Mittenwald:

1 specimen, leg. Osthelder (ZSM); Regensburg: 1 specimen (ZSM); Ruhpolding: 1 specimen (ZSM);

Bamberg: | specimen, leg. Garthe (ZSM). Poland, Katy, distr. Zamosc: 14, 4.viii.1978, leg. Buszko

(ZMUC). Denmark, Lolland, Rodbyhaven: 1 specimen, vii 2010, leg. Larsen ( ZMUC). Finland, Karis-

lojo: 14, 12.viii.1967, leg. et coll. Krogerus.

Ochromolopis zagulajevi Budashkin & Sachkov, 1991

Italy, Lucania, Mt. Pollino, 39°50’N, 13°33’E: 45, 3.x.2010, 26.vii.2011, leg. Skou (ZMUC). Croatia,

Fiume: 24, 1853 (NMW); Istria, Belavici, Marcana: 14, 69, 8.-14.ix.2008, leg. Mey (ZMHB; SDEI);

Dalmatia, env. of Selce: 48, 8.—15.viii.1989, leg. Gestberger (SDEI). Macedonia, Stari Dojran: 19, 2.-

10.vi.1956, leg. Klimesch (ZSM); Drenovo near Kavadar: 14, 10.-20.vi.1956, leg. Klimesch (ZSM).

Albania, Kula Ljums [Kula e Lumes]: 14 (NMW); Korab: 14, 23.-31.vii.1918 (NMW); Gjalica Ljums

[Mail i Gjalices]: 24, 17.-16.vi.1918 (NMW; SDEI); Sisevo near Üsküb: 14, 11.v.1918 (NMW). Greece,

Peloponnese, Zachlorou near Kalavrita: 19, 1.-14.vii.1959, leg. Noack (LMAD); Karia: 1 specimen,

12.vii.1974, leg. et coll. Arenberger; Itea/Desfina: 24, 29.v.2006, leg. et coll. Schmitz. Bulgaria, Pirin,

Sandanski: 94, 72, 17.-31.v.2010, 28.iii—4.iv.2011, leg. Savenkov (coll. Roweck; SDEI). Turkey, Zei-

toon: 19 (ZMHB); Sivas, darende Günpinar: 14, 18.x.1986, leg. Moberg & Hilman (ZMUC); env. of

Urgiip: 1 specimen, 24.vi.1969, leg. et coll. Arenberger; 50km N Tarsus: 1 specimen, 19.v.1969, leg. et

coll. Arenberger; Yalova at Sea of Marmara: 1 specimen, 11.v.1969, leg. et coll. Arenberger; Djanik, E of

Terme: 14, 6.v.1969, leg. Glaser (SMNK); Erzurum: 14, 17.ix.1993, leg. Fibiger (ZMUC); Agri, 23km

W Dagubayazit: 14, 5.ix.1993, leg. Fibiger (ZMUC). Ukraine, Crimea, Dobroje Krasnolesje: 484, 59,

18.v.1983, 5.vii.1986, leg. Zagulajev (ZIN; SDEI); Crimea, Karadag: 14, 22.v.1984, leg. Zagulajev (ZIN).

Russia, Kaukasus, Kabardino-Balkarski nature reserve, 35km SE of Elbrus: 104,29, 9.-13.vii.1990, leg.

Jalava (FMNH; SDEI). Georgia, Tbilissi: 14, 30.v.-1.vi.1971, leg. Muche. Armenia, Geghard, 40km

E of Erewan: 14, 26.-27.vii.1976, leg. Kasy & Vartian (NMW). Iran, Keredj, Elburs Mountains: 26,

19, 15.iv.1936, leg. Brandt (NHRS); Balochistan, Bendar Tschahabahar: 14, 1937, leg. Brandt (NHRS).

Ochromolopis kaszabi Gaedike, 1973

Russia, SW-Altai, Kuragan valley, 15km S Katanda, 1200m: 64, 23.-25.vii.1983, Exp. K. Mikkola,

H. Hippa & J. Jalava (FMNH; SDEI); Buryatia, 53°40’N, 109°00’E, Svyatov Nos pns. Monahovo,

460m: 2, 13.vii.1996, leg. J. Jalava & J. Kullberg (FMNH); Sibiria, Tschita, reka [=river] Ingoda: 14,

27.vii.1997, leg. I. Kostjuk (ZMHB); Primorskij kraj, Pogranitschnyj Rajon, Barabasch-Levada: 14, 59,

14.vii.1989, leg. S. Zinjov (ZIN); Chazanskij Rajon, Kedrovaja pad’: 24, 29, 25.vii., 1., 2.viii.1988,

leg. S. Zinjov; 1 specimen, 15.vii.1974, leg. Ermolajev (ZIN); Chazanskij Rajon, Rezanovka: 24, 29,

22., 23.viii.1982, leg. S. Zinjov (ZIN); Chazanskij Rajon, 3km SE Andrejevka: 54, 59, 21., 24.vii.,

7., 11., 15.viii.1985, 12,viii.1984, leg. S. Zinjov (ZIN); Chazanskij Rajon, 7km N Zanadvorovka: 59,

14.viii.1984, leg. S. Zinjov (ZIN); Ussurijskij Rajon: 25, 49, 3., 12., 19.vii., 14., 19.viii.1982, leg. S.

Zinjov (ZIN); Ussurijskij Rajon, surround of Ussurijsk: 34, 69, 28.vi., 7., 8., 28.vii., 3.ix.1983, leg. S.

Zinjov (ZIN); Ussurijskij Rajon, 20km E of Ussurijsk: 54, 49, 20.viii.1980, 22.viii.1981, 9.vii.1984, 3.,

13.v11.1985, leg. S. Zinjov (ZIN). Mongolia, Bulgan aimak, 7km NW of Somon, Chanzar-galant, 1350m:

164, 22.vii.1968, leg. Z. Kaszab (HNHM; SDEI).

Nota Lepi. 37(1) 2014: 63-65 | DOI 10.3897/n1.37.7956

Description of the female of Ethmia cribravia Wang and Li 2004

(Lepidoptera, Elachistidae, Ethmiinae)

Dmitry F. SHOVKOON!, TATIANA A. TROFIMOVA’

1 Laboratory of Biogeography and Monitoring of Biodiversity of Steppe Institute, Ural Branch of Russian Academy

of Sciences, 460000 Orenburg, Pionerskaya st, 11; shovkoon@mail.ru

2 Laboratory of Animal Systematics and Faunistics, Samara State University, 443011 Russia, Samara, Academica

Pavlova st. 1; apame@mail.ru

http://zoobank.org/067106C4-57C9-458D-9595-22532E43388E

Received 4 March 2014; accepted 7 April 2014; published: 15 June 2014

Subject Editor: Jadranka Rota

Abstract. The previously unknown female of Ethmia cribravia Wang and Li, a species known from

Yunnan, China, is described and illustrated with colour photographs of the habitus, as well as images of

genitalia mounted on slides. The species is sexually dimorphic, with the male having prominent androco-

nial scales on the hindwing.

Pestome. B crarbe HPHBOAATCH ONHCAHHe paHe HeH3BeCTHOË camKU Ethmia cribravia. BnepBbie aHbl

IBeTHbIe WIWIOCTpalHu Oadouek, Tak Xe aHbI YepHO-OesIbIe POTOrpaADAA TEHHTANBHBIX CTPyKTyp Camila

N CAMKH.

Introduction

Ethmia cribravia was described by Wang and Li (2004) ten years ago, based on three male

specimens from Yunnan Province of China. In the collection of the Hungarian Natural History

Museum (Budapest) we found two male and two female specimens of this little-known species.

E. cribravia is peculiar as it displays remarkable sexual dimorphism. The male has a tuft of

long, piliform androconial scales on the anal margin of the hindwing (Fig. 1). A similar degree

of sexual dimorphism is known from only one other Ethmia species — the male of the African

species Ethmia melanocrates Meyrick, 1923 has similar androconial scales on the anal margin

of the hindwing (Mey & Shovkoon 2014: fig. 14).

Abbreviations

HNHM Hungarian Natural History Museum, Budapest.

64 JADRANKA Rota et al: Description of the female of Ethmia cribravia Wang and Li 2004

Figures 1-4. Ethmia cribravia. 1. Male, China, Yunnan Diqing Tibetan (HNHM). 2. Female, China,

Yunnan Diqing Tibetan (HNHM). 3. Male genitalia: a general view and b phallus; Shovkoon praeparavit,

prep. Ne 219 (HNHM). 4. Female genitalia: a general view, b bursae, ce signum; Shovkoon praeparavit,

prep. Ne 220 (HNHM).

Ethmia cribravia Wang and Li 2004

Figs 14

Material. 27, 29 China, Yunnan Diging Tibetan Auf. Pref. Tiger Leaping Gotge, SE Slope at Sean’s Guesthouse,

2000-2500 m N 27°16.113’, E 100°10.233’ 9-12.vi.2008, leg. B. Benedek. (HNHM).

Description of female. (Fig. 2) Length of forewing 29.5-30.5 mm. In general coloration

and pattern of forewing essentially as male. Hindwing lacking the dark field in the centre and

piliform androconial scales on anal margin of hindwing, typical of male.

Nota Lepi. 37(1) 2014: 63-65 65

Female genitalia (Fig. 4). Papillae anales elongated, setose. Eighth tergite medially with mem-

branous incision. Posterior apophyses slender, as long as papillae anales; anterior apophyses

wedge-shaped, distally pointed (Fig. 4a). Posterior part of antrum sclerotized with armed scle-

rotized thorns. Ductus bursae as long as abdomen, with 5—7 coils, bursa copulatrix spherical (Fig.

4b). Signum very large, cruciform, evenly covered with small teeth (Fig. 4c).

Distribution. The collecting site of the specimens examined by us is only 20 kilometers

north-east of the type locality, both sites in Yunnan Province of China.

Taxonomic notes. The moth habitus and the structure of the male (Fig. 3) and female gen-

italia (Fig. 4) suggest a close relationship with E. dehiscens Meyrick, 1924, and the species is

therefore placed in the dehiscens species-group sensu Sattler (1967).

The similarity in the appearance of the androconial scales of E. cribravia and the African

species E. melanocrates is puzzling. The two species do not belong in the same species-group

sensu Sattler (1967). Based on the morphology of the genitalia of E. melanocrates (Mey &

Shovkoon 2014: figs 17, 18), this species belongs to its own independent species-group, the

members of which are known only from Kenya, Namibia, and South Africa and are not present

in the Palaearctic region.

Additionally, these two species differ in their external appearance. The costal half of the fore-

wing of E. melanocrates is suffused with yellow scales, and the basal half with a complex pattern

formed by dark and light fields; tiny black marginal dots are present; the cilia are pale grey, with

black scales at the tip of the forewing; the hindwing is yellow with yellow cilia; and the abdomen

and thorax are grey-yellow (Mey & Shovkoon 2014: figs 14, 15).

It is very interesting that, in spite of such differences in morphology and distribution, the an-

droconial scales of these two species are similar to such an extent. In both species the piliform

androconial scales are located on the anal margin of the hindwing starting from the An, vein

and they are as long as the width of the hindwing.

Acknowledgements

We are very grateful to Dr. Zsolt Balint (Budapest) for his help in studying museum collections of Hun-

garian Natural History Museum.

References

Mey W, Shovkoon DF (in press). Synopsis of the Ethmiinae (Lepidoptera, Gelechioidea: Depressariidae) of

the Afrotropical Region and descriptions of new species. Tijdschrift voor Entomologie 157.

Sattler K (1967) Ethmiidae. In Amsel HG, Gregor F, Reisser H (Eds). Microlepidoptera Palaearctica 2

(1+2). — Georg Fromme, Wien, xviii + 185 pp., 106 pls.

Wang S, Li H (2004) Two new species of Ethmia Hübner from China Lepidoptera: Elachistidae: Ethmi-

inae. Entomological News 115 (3): 135-138.

Nota Lepi. 37(1) 2014: 67-74 | DOI 10.3897/n1.37.7935

A remarkable new species of the genus Catatinagma Rebel, 1903

(Lepidoptera, Gelechiidae) from Turkmenistan

OLEKSIY V. BIDZILyYA'

1 Kiev National Taras Shevchenko University, Zoological Museum, Volodymyrska str., 60, MSP 01601, Kyiv,

Ukraine; bidzilya@univ.kiev.ua

http://zoobank.org/8A092C1D-38A1-47D9-B56C-9513184E4F6D

Received 31 January 2014; accepted 29 April 2014; published: 15 June 2014

Subject Editor: Lauri Kaila

Abstract. A new highly specialized Catatinagma Rebel, 1903 species is described from Turkmenistan.

Both sexes have completely reduced hindwings and strongly reduced forewings. The adults are active

in February, jumping amongst Carex physodes M. Bieb. and being associated with rodent burrows. The

new species is similar to Metanarsia trisignella Bidzilya, 2008, in the male genitalia. Both species are

placed here provisionally in Catatinagma Rebel, 1903, and their position within Apatetrini is briefly

discussed. The adult and the genitalia of both sexes are illustrated, and the behaviour of the new species

is described.

Introduction

As aresult of my study of material deposited in the Zoological Institute of the Russian Acade-

my of Sciences (Russia, Sankt-Petersburg, ZIN), a very remarkable narrow-winged species of

Gelechiidae with prominent frontal process from Repetek Nature Reserve (SE Turkmenistan)

was discovered.

As it turned out after a detailed examination, the species was an undescribed member of the

subfamily Apatetrinae, tribe Apatetrini (Karsholt et al. 2013) but its generic assignment was

unclear. A well-developed beak-shaped frontal process on the head and stenoptery in both sexes

with fully reduced hindwing were recognized as external morphological specializations of the

new species. The male genitalia of the new species resemble those of Metanarsia trisignella

Bidz., but the female genitalia differ in the shape of the signum. Both species are placed provi-

sionally in Catatinagma Rebel, 1903, until a more appropriate place can be found.

In addition to strongly modified wings, the new species has a very unusual behaviour. The

adults hide in the burrows of rodents in cold weather. This unique life style and the rather re-

markable morphology of this species justify its description in advance of a broader taxonomic

revision of the Apatetrini.

68 BıpzıLyA: A remarkable new species of the genus Catatinagma Rebel, 1903...

Catatinagma stenoptera Bidzilya, sp. n.

http://zoobank.org/83 160C15-A639-4756-A9C8-3 1C73BC82F9D

Material. Holotype, 4, Karakum desert, Repetek reserv[e], Carex, 3.11.1983, Krivokhatsky (gen. slide 55/11) (ZIN).

Paratypes: 1 <, same data as holotype (gen. prep. in glycerol); 1 4, same data as holotype, but 18.ii.1983, trap Rhom-

bomys, night (ZIN-00002); 1 ©, SAME data, but 15.11.1983, trap Rhombomys (gen. slide 56/11) (ZIN-00005); 1 à,

2 ©, Repetek, SE Karakum, Turkmenia, trap Rhombomys, 25.ii.1983 (ZIN-00004 3, ZIN-00003 ©, © gen. prep. in

glycerol) (all ZIN).

Description. Adult (Figs 1-3). Wingspan 9.1-10.9 mm. Head smooth-scaled, whitish-brown,

with prominent beak-shaped, pointed process, that sometimes bears additional small dorsal

knob arising from middle; frons medially with depression resembling excavator bucket with

three short tooth-shaped projections at bottom; labial palpus weakly up-curved, outer surface

black with white basal and apical belts, inner surface white, segment 3 nearly straight, acute;

segment 2 about 1.5 times width and nearly 2.5 times length of segment 3; proboscis reduced;

antenna brown with very narrow whitish ring at base of each segment, pecten with numerous

long white hair-like scales; forewing strongly narrowed after 4, nearly filiform in distal half,

light brown, mottled white along margins, cilia white; hindwing vestigial.

Abdomen (Figs 4—7). Male sternite VIII rectangular, without modification. Female tergite

VII nearly twice length of other abdominal segments, rectangular, slightly broader than long,

densely covered posteriorly with short hair-like scales, sternum VII rectangular without mod-

ification. Sternite I+II of both sexes sub-quadratical, with pair of distinct long venulae and

well-developed apodemes, tergite I distinctly broader than long, strongly edged, anterolateral

margin rounded, posterior margin straight.

Male genitalia (Fig. 8). Uncus reduced to trapezoidal lobe with inward folded edges;

gnathos absent; tegumen narrow, considerably broader than long, posterior edge strongly

sclerotized; cucullus digitate, moderately broad, apex rounded, densely haired; sacculus

flat, about 3/4 length of cucullus and slightly broader, posterior margin straight with two

or three small teeth; transtilla lobes well developed; vinculum narrow, band-shaped, termi-

nating in short rounded saccus; phallus longer than cucullus, with tapered lateral processes,

basal half sclerotized dorsally, distal half sclerotized mainly laterally, apex beak-shaped,

base bifurcated.

Female genitalia (Fig. 9). Papilla analis rounded, with straight basal edge, densely covered

with short setae except for patch of long hair-like setae arising from dorsal margin; apophyses

anteriores about one-half length of apophyses posteriores, straight, terminally curved; tergite

and sternite VIII extremely narrow, strongly sclerotized, ribbon-shaped; lateral part of segment

VIII evenly sclerotized, band-shaped; ostium rounded, opening near anterior edge of sternite

VIII; antrum short, funnel-shaped; ductus bursae long, membranous, posterior half thin, anteri-

or half moderately wide; corpus bursae globular; signum paired, with long spines arising from

rounded plate.

Diagnosis. The new species is easily recognizable both externally and in the genitalia char-

acters. For details see the Discussion.

Distribution. SE Turkmenistan (Repetek Nature Reserve).

Derivation of name. The specific name refers to the extremely narrowed forewing, the most

characteristic feature of this species.

Nota Lepi. 37(1) 2014: 67-74 69

Figs 1-9. Catatingama stenoptera sp. n. 1. Adult, holotype. 2. Head, lateral view. 3. Head, view from

above; 4. Male sternum VIII (gen. slide 55/11). 5. Female tergum VII (gen. slide 56/11). 6. Male sternum

IH (gen. slide 55/11). 7. Male tergum I (gen. slide 55/11). 8. Male genitalia (gen. slide 55/11). 9. Female

genitalia (gen. slide 56/11).

70 BiDziLYA: A remarkable new species of the genus Catatinagma Rebel, 1903...

Biology. The new species is hitherto only known from the Repetek Nature Reserve, SE

Turkmenistan. The adults were collected from 3" to 25" of February. According to field obser-

vations by Viktor A. Krivokhatsky, who collected the type-series, the adults were active during

the warm days when they were seen jumping on the sand. They have also been collected by

sweeping amongst Carex physodes M. Bieb. (Cyperaceae). This plant is most likely the host

for this species, although the preimaginal stages have not yet been found. The adults have also

been observed and collected in the burrows of the great gerbil (Rhombomys opimus (Lichten-

stein, 1823)) and the long-clawed ground squirrel (Spermophilopsis leptodactylus (Lichten-

stein, 1823)) (Mammalia, Rodentia, Muridae, Sciuridae). The moths go deeply into the burrows

at night and when there is frost in the daytime.

Discussion

Wing reduction and biology. Within the family Gelechiidae species with different degrees

of wing reduction are known in eleven genera, but are most common in Megacraspedus

Zeller, 1839, Ephysteris Meyrick, 1908, Kiwaia Philpott, 1930, and Sattleria Povolny, 1965

(Sattler 1991). The forewing transformation, for the majority of such Gelechiidae species,

resulted in forewing reduction of length and width in the apical one-third. As a result the

forewings become “lanceolated”: their basal half remains nearly of usual width, whereas

the distal portion is considerably narrowed towards the apex. According to the classification

proposed by Sattler (1991) such species must be called “brachypterous”, whereas the species

described here has to be treated as “stenopterous”, having the forewing strongly reduced

in width without any significant reduction in length. Brachyptery and other cases of wing

reduction are mainly restricted to the female sex. Megacraspedus dolosellus (Zeller, 1839),

Eulamprotes libertinella (Zeller, 1872) and all species of the genus Sattleria are some ex-

amples of brachypterous Palaearctic Gelechiidae. Wing reduction in both sexes is a rather

rare phenomenon that is known in about 25 species from different families of Lepidoptera

world-wide (Karsholt & Sattler 1998; Sattler 1991). In Gelechioidea real stenoptery is only

known in the female of Pleurota marginella (Denis & Schiffermüller, 1775), Oecophoridae,

and previously has not been observed within Gelechiidae, making C. stenoptera sp. n. the

first case of stenoptery in both sexes in that family.

Wing reduction in both sexes characterizes the species that inhabit mainly small oceanic

islands, costal localities (e.g. dunes) and high mountain area with continuous strong winds

(Heppner 1991; Sattler 1991; Karsholt & Sattler 1998). Ephysteris brachyptera Karsholt &

Sattler, 1998, from Madeira and at least two undescribed Ephysteris species from the Tuva

Mountains (Karsholt & Sattler 1998) and Mongolia (Bidzilya in press) are examples of

brachyptery in both sexes in Palaearctic Gelechiidae. Brachyptery in females is much more

common and observed in addition to the above cases, in a large number of species with imagos

active during the cold season. C. stenoptera sp. n. undoubtedly falls into this group of “winter”

moths. The adults were found in winter time in the Karakum desert. Among Turanian Gele-

chioidea species with winter activity of the imago are known only in Ethmiidae: Dasyethmia

hiemalis Danilevsky, 1969, from the sands of south-eastern Kazakhstan (middle course of the

Chu river) flies in the daytime in January. The males of this species are densely haired, the

females are unknown but probably wing-reduced (Shovkoon 2010). Cheimoptena pennigera

Nota Lepi. 37(1) 2014: 67-74 7

Danilevsky, 1969 (Geometridae) is another winter moth described from the south-eastern Ka-

zakhstan but also known from Repetek (Krivokhatsky 1985c). The female of this species is

apterous, the males are capable of flight and densely haired, as an adaptation for activity under

low temperatures.

Adults of C. stenoptera are flightless, they move by jumping. The jumping and/or running

may be considered as a regular way of locomotion for many flightless species, that mainly

occurs in habitats with sparse vegetation and exposed soil, although jumping is quite com-

mon in many fully winged species capable of flight (Sattler 1991). It is unknown whether

jumping is the only way of locomotion in C. stenoptera or whether the adults can also walk

or run. However, it is clear that C. stenoptera is a quite mobile species: both sexes were found

deep in rodent burrows, where they hide at night and during frost also in the day time. The

association of C. stenoptera with rodent burrows is the most interesting behavioural phenom-

enon of this species. The great gerbil and the long-clawed ground squirrel are quite common

in the sand deserts of Turkmenistan. The first species is considered ecologically one of the

most important mammals, whose activity (mainly digging holes) considerably affects the

local landscape. Many invertebrate species are associated with the great gerbil and inhabit

its burrows. According to observations in the Repetek Nature Reserve (Krivokhatsky 1985a)

about 345 animal species, mainly insects, were found in the burrows of great gerbil and

other rodents. The Coleoptera (121 species of which 54 are obligate residents), Hymenop-

tera (50 species / 5 obligate) and Hemiptera (29 species / 6 obligate) predominate amongst

the insects. The Lepidoptera with 15 species occupy the fourth position. Only two species

- Calycobathra calligoni Sinev, 1979, and Asclenia decolorella Sinev, 1980 (both Cosmop-

terigidae) - overwinter in the burrows of the great gerbil and are considered as permanent

residents (Krivokhatsky 1985b). C. stenoptera is the third species strongly associated with

the great gerbil’s burrows. In contrast to the Momphidae species, the adults of C. stenoptera

use rodent burrows as a refuge during the period of their activity in winter time, but not for

overwintering in them.

The host plant of C. stenoptera is unknown. The adults have been observed amongst Carex

physodes, but Haloxylon spp., Kochia spp. (Chenopodiaceae) and Ephedra strobilacea Bunge

(Ephedraceae), which are common in the habitats of C. stenoptera, are also possible candidates

for the host plant of this species.

The function of frontal processes in Gelechiidae remains unclear. It was suggested that spe-

cies pupating under ground use the frontal process to bore through the soil after emerging

from the pupa. However, as it was correctly noted by Sattler (1976), many species in the same

habitats manage perfectly well without such modifications. Frontal modifications occur inde-

pendently in several gelechiid genera from different subfamilies and tribes, such as Ornativalva

Gozmany, 1955 (Anomologinae), Athrips Billberg, 1820 (Gelechiinae), Caulastrocecis Chré-

tien, 1931 (Apatetrinae) and others. In the Apatetrinae the frontal modifications are common

and were observed in Palaearctic Catatinagma as well as in the South African genera Cerof-

rontia Janse, 1951, and Rhynoceros Bidzilya & Mey, 2011. As most species with frontal mod-

ifications are restricted to arid areas, one may suspect some relationship of this modification to

occurrence in arid habitats, but not as an adaptation to the cold season.

The Repetek Nature Reserve, where C. stenoptera has been found, represents the south-

ern sand deserts of the Turanian province (Karakum desert). The area is characterized by an

72 BiDziLYA: A remarkable new species of the genus Catatinagma Rebel, 1903...

arid continental climate with hot summers and cold winters. The mean annual temperature

is +16.3°C, with the average summer temperature ranging +29-33°C (with a record high of

+50.1°C); temperatures below freezing are usually observed from December to February (with

a record low of -31°C). The average precipitation is 117 mm per year with a maximum in

winter and spring (Zapovednik Repetek 1990). The climate of Repetek is in general similar to

that of the type localities of D. hiemalis and Ch. pennigera, but the winter is milder. Although

these three winter species occur under very similar climatic conditions, C. stenoptera has rather

different morphological and biological adaptations for the cold season. The phenomenon of

wing reduction in cold season moths was discussed in detail by Sattler in his review of the wing

reduction in Lepidoptera (Sattler 1991).

Although our current knowledge of the distribution, habitat preferences, host plants and

behaviour of C. stenoptera is rather limited it seems clear that the regular occurrence of C.

stenoptera in the burrows of the great gerbil and the long-clawed ground squirrel is a perma-

nent adaptation for surviving under a low temperature, at least in the Repetek.

Systematics

The monotypic genus Catatinagma with the only included species C. trivitellum Rebel, 1903,

was considered for a long time as a synonym of Apatetris Staudinger, 1879. Its status has been

recently revised and it was recalled from synonymy, whilst another monotypic genus, Coloptil-

ia Fletcher, 1940, has been synonymized with Catatinagma (Junnilainen & Nupponen 2010).

According to Junnilainen & Nupponen (2010), the head with a strongly developed frontal pro-

cess within a crater-like depression and a short, nearly straight labial palpus are considered

as external characteristic features of Catatinagma. The male genitalia are characterized by a

sacculus that is strongly separated from the valva, a reduced gnathos and membranous uncus.

A paired brush-shaped signum is typical for the female genitalia of Catatinagma (Junnilainen

& Nupponen 2010).

The genus Catatinagma in this new concept comprises three species: C. trivitellum Rebel,

1903, C. kraterella Junnilainen & Nupponen, 2010 and C. conchylidella (Hofmann, 1898)

(type-species of Coloptilia). Whilst it is evident that the first two species are congeneric, the

proximity of C. conchylidella to them, and consequently the synonymy of Coloptilia with Cat-

atinagma, remains in doubt due to considerable differences between the male genitalia of C.

trivitellum and C. conchylidella (see Junnilainen & Nupponen 2010: 6, pl. 2).

The male genitalia of C. stenoptera resemble those of Metanarsia trisignella Bidzilya,

2008, having a short digitate cucullus in combination with a short, apically serrated sacculus.

The female genitalia of both species differ in the shape of the signum: triple whisk-shaped

in M. trisignella and paired, covered with strong spines in C. stenoptera. M. trisignella was

described in Metanarsia Staudinger, 1871, based on the close similarity of its male genitalia

to the other members of this genus, although a reduced gnathos and triple whisk-shaped sig-

num as well as a prominent frontal process are characteristic features of Catatinagma, but not

Metanarsia (Bidzilya 2008). C. trivitellum is the second species in addition to M. trisignella

which appears to be intermediate between Metanarsia and Coloptilia according to the current

diagnosis of these genera. It should be noted that the male genitalia of both species are more

like those of C. conchylidella than those of C. trivitellum and C. kraterella. Despite differ-

Nota Lepi. 37(1) 2014: 67-74 73

ences in the shape of the signum, both species seem to be closer relatives of each other than

any other genera of Apatetrini. The recent attempts of finding an appropriate position for such

“problematic” species within the tribe Apatetrini based on traditional morphological charac-

ters usually resulted either in their provisional association with collective genera of unclear

definition (Sakamaki 2000; Nel & Varenne 2012) or in the establishment of new genera for

the most exceptional cases (Bidzilya & Mey 2011). The possibility cannot be excluded that

both species require a separate genus. However, it seems more appropriate at the present stage

to place them in Catatinagma (Catatinagma trisignella (Bidzilya, 2008) comb. n.)), until a

global revision of Apatetrini supported by DNA-studies can be provided.

Acknowledgements

I express my gratitude to Dr Sergei Yu. Sinev (ZIN) for his assistance during my work with the collec-

tion and Dr Viktor A. Krivokhatsky (ZIN), who kindly provided information about behavior of the new

species. I am very grateful to Dr Klaus Sattler (the Natural History Museum, London), Jaakko Kullberg

(Finnish Museum of Natural History) and an anonymous reviewer for their valuable comments and lin-

guistic correction of an earlier version.

References

Bidzilya OV (2005) A review of the genus Metanarsia Staudinger, 1871 (Gelechiidae). Nota lepidopter-

ologica 27 (4): 273-297.

Bidzilya OV (2008) New species and new records of the genus Metanarsia (Lepidoptera, Gelechiidae).

SHILAP Revista de Lepidopterologia 36 (144): 531-538.

Bidzilya OV, Mey W (2011) Gelechiidae. In Mey W (ed). Basic pattern of Lepidoptera diversity in south-

western Africa. Esperiana Memoir 6: 197-216.

Junnilainen J, Nupponen K (2010) The gelechiid fauna of the southern Ural Mountains, part I: description

of seventeen new species (Lepidoptera: Gelechiidae). Zootaxa 2366: 1-34.

Heppner JB (1991) Brachyptery and aptery in Lepidoptera. Tropical Lepidoptera 2 (1): 11-40.

Karsholt O, Sattler K (1998) A new brachypterous Ephysteris Meyrick from the island of Madeira (Lepi-

doptera: Gelechiidae). Entomologist’s Gazette 49: 35-47.

Karsholt O, Mutanen M, Lee S, Kaila L (2013) A molecular analysis of the Gelechiidae (Lepidoptera,

Gelechioidea) with an interpretative grouping of its taxa. Systematic Entomology 38: 334-348.

Krivokhatsky VA (1985a) Nasekomye norovykh konsortsiy peschanych pustyn’ Srednei Asii [Insects of

burrows consortium from the sandy deserts of Middle Asia]. Thesis to acquire the degree of candidate

of biological sciences. Speciality 03.00.09 — Entomology. Leningrad, 20 pp. [in Russian].

Krivokhatsky VA (1985b) On the history of the forming of the nidicolous entomofauna of sandy deserts

of Middle Asia. Entomologicheskoe obozrenie 64 (4): 696-704 [in Russian].

Krivokhatsky VA (1985c) Nasekomye Repeteka. Kadastr vidov [Insects of Repetek. Cadastre of species].

Ylym, Ashhabad. 72 pp. [in Russian].

Nel J, Varenne T (2012) Description d’Apatetris (s.l.) mediterranella sp. n. du littoral méditerranéen de

France et d’Italie (Lepidoptera, Gelechiinae, Apatetrini). Nota lepidopterologica 35 (1): 27-32.

Sakamaki Y (2000) Japanese species of the genus Apatetris (Lepidoptera, Gelechiidae). Tijdschrift voor

Entomologie 143: 211-220.

Sattler K (1976) A taxonomic revision of the genus Ornativalva Gozmäny, 1955 (Lepidoptera, Gelechii-

dae). Bulletin of the British Museum (Natural History), Series Entomology 34 (2): 87-152.

Sattler K (1991) A review of wing reduction in Lepidoptera. Bulletin of the British Museum (Natural

History) (Entomology) 60 (2): 243-288.

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Shovkoon D (2010) New and little-known Ethmiidae (Gelechioidea) from Central Asia. Nota lepidopter-

ologica 33 (1): 135-154.

Zapovednik Repetek (1990) Zapovedniki Srednei Asii 1 Kazakhstana [Natural Reserves of Middle Asia

and Kazakhstan]. In: Sokolov VE, Syroechkovsky EE (Eds) Zapovedniki SSSR. Mysl’, Moskow.

200-216. [in Russian].

Nota Lepi. 37(1) 2014: 75-90 | DOI 10.3897/n1.37.7966

Wing pattern morphology of three closely related Melitaea

(Lepidoptera, Nymphalidae) species reveals highly inaccurate

external morphology-based species identification

JURE JuGovic!”, TONI KOREN?

1 University of Primorska, Faculty of mathematics, natural sciences and information technologies, SI-6000

Koper, Glagoljaska 8, Slovenia; jure.jugovic@zrs.upr.si

2 University of Primorska, Science and Research Centre, Institute for Biodiversity Studies, S1-6310 Izola, Giordana

Bruna 6, Slovenia; koren.tonil@gmail.com

http://zoobank. org/29B3CAES5-7E75-416E-9555-D85F 28 1SBBAO

Received 22 November 2013; accepted 5 May 2014; published: 15 June 2014

Subject Editor: Zdenék F. Fric

Abstract. Wing morphology of the three closely related species of Melitaea — M. athalia (Rottemburg,

1775), M. aurelia (Nickerl, 1850) and M. britomartis Assmann, 1847 — co-occurring in the Balkans (SE

Europe) was investigated in detail through visual inspection, morphometric analysis and multivariate

statistical analysis. Results are compared to recent phylogenetic studies, searching for concordant patterns

and discrepancies between the two approaches. The morphology of the genitalic structures is also com-

pared with the results of the other two approaches. The main conclusions are as follows: (1) small albeit

significant differences in wing morphology exist among the three species and (2) while the structure of

male genitalia and phylogenetic position of the three species are concordant, they are (3) in discordance

with the wing morphology. The present study represents another example where identification based on

external morphology would lead to highly unreliable determinations, hence identification based on phy-

logenetic studies and/or genitalia is strongly recommended not only for the three studied species but also

more broadly within the genus. Furthermore, we show that some of the characters generally used in the

identification of these three Melitaea species should be avoided in future.

Introduction

The genus Melitaea Fabricius, 1807 (Lepidoptera: Nymphalidae) as it is known today (Len-

eveu et al. 2009), comprises approximately 80 species, all restricted to the Palaearctic region.

A recent phylogenetic analysis showed that the genus can be divided into two clades (‘Meli-

taea clade’ and ‘Didymaeformia clade’) with high branch support from both maximum likeli-

hood and Bayesian analyses (Leneveu et al. 2009). Within the ‘ Melitaea clade’ five subclades

have been recognized (sensu Leneveu et al. 2009): ‘cinxia’, ‘diamina’, ‘arcesia’, ‘minerva’

and ‘athalia’ groups. The latter subclade comprises fourteen species, three of which are pres-

ent in the area of north-western Balkans: Melitaea athalia (Rottemburg, 1775), Melitaea au-

relia (Nickerl, 1850) and Melitaea britomartis Assmann, 1847. These three species belong to

two different monophyletic groups within the subclade ‘athalia’, one comprising M. aurelia

and five other species, and the other M. athalia, M. britomartis and six other species (Leneveu

et al. 2009). The split between the latter two monophyletic groups has been estimated to have

76 JUGOVIC & KOREN: Wing pattern morphology of three closely related Melitaea...

occurred about 11 million years ago (Serravallian period) while the subsequent speciation oc-

curred during the Messinian Period (5.3—7.1 Mya) (Leneveu et al. 2009), when the continuous

decrease of temperature coincided with the aridification of the climate in Eurasia and a subse-

quent expansion of grassland of C4 plants (e.g. Maki et al. 2003; Quade et al. 1995). Although

the separate evolution of those species has been in progress for a long period of time, the

clear genetic differentiation (Batori et al. 2012a) is not reflected in their external morphology

(Tolman & Lewington 2008).

The external morphology of the three species is highly similar, rendering the accurate iden-

tification based on external morphology questionable (Tolman & Lewington 2008). There

are several morphological characters that have been proposed for species identification of M

athalia, M. aurelia and M. britomartis: the coloration of the marginal line of the hindwing

underside; the spacing of the marginal, submarginal, postdiscal and discal line on the forewing

upperside; and the colour of the submarginal spots on the hindwing underside (Tolman & Lew-

ington 2008). The high variability of those characters, however, sometimes prevents proper

identification (Tolman & Lewington 2008). The genitalia of these species are highly species

Tamutis 2009). Nevertheless, even when genitalia based identification is used, only males can

be unambiguously recognized to a species rank, whereas this is not always possible in females

(e.g. see Urbahn 1952).

Melitaea athalia is a trans-European species (except for south-western Europe, where Mel-

itaea celadussa Fruhstorfer, 1910, a former subspecies of M. athalia occurs (Leneveu et al.

2009)). M. aurelia and M. britomartis are absent from the majority of SW Europe (including

the Iberian and Apennine peninsulas). M. britomartis is absent also from most of Central Eu-

rope being present only in eastern parts of Europe. A part of its distribution, however, reaches

the NW Balkans — Slovenia, Croatia and Bosnia and Herzegovina (Koren & Jugovic 2012)

— where these three species live in sympatry. In this area the distribution of M. britomartis is

separated from the center of its distribution (Tolman & Lewington 2008).

The ecology of the three species in the area of NW Balkans (Slovenia) is similar in terms

of flight period, altitudinal distribution and habitat requirements including foodplant (Koren

& Jugovic 2012; Verovnik et al. 2012). Since their habitat requirements and foodplants partly

overlap, they often occur in the same habitat (Batori et al. 2012a; Koren & Jugovic 2012).

Morphological traits can reflect either historical isolation and/or local adaptation despite re-

current gene flow (Alexandrino et al. 2005). Some taxa remain morphologically unidentifiable

despite detailed morphometric analysis, hence a consideration of a large set of morphometric

characters from different anatomical regions may greatly increase the chances of revealing tax-

onomic differences within seemingly cryptic or morphologically highly variable species (e.g.

Jugovic et al. 2011).

We wanted to check the concordance in the amount of morphological and genetic differ-

ences among the three closely related species. For this purpose a morphometric approach with

subsequent multivariate statistical analysis was used in the M. athalia complex for the first

time. We evaluated the following hypotheses: (1) the genetic divergence of the three species is

accompanied by corresponding morphological differences exceeding the intra-species variabil-

ity and (2) the level of accompanying morphological differences is concordant with the level

Nota Lepi. 37(1) 2014: 75-90 77

of molecular distance between the taxa, meaning that M. aurelia should be the most distant in

morphological space from the other two species.

Materials and methods

Samples and species identification

Samples of the three species of Melitaea were collected over the last 40 years all over the NW

Balkans (see Koren & Jugovic 2012 for the list). Altogether, samples from 56 localities were

collected, sample sizes ranging from 1 to 12 for males and 1 to 3 for females. The genitalia of

each specimen were isolated using the standard procedure for genitalia isolation (see Koren

& Jugovic 2012 for details). Specimens were attributed to a species according to the genitalia

Using these methods we were able to identify all of the collected males. A total of 42 males and

24 females of M. athalia, 29 males and 6 females of M. aurelia and 24 males and 4 females of

M. britomartis were used in further morphometric analysis.

Separation of these three species is possible using male genitalia as follows (see also Urbahn

1952): 1. uncus absent -> M. aurelia; 2. uncus present, long and slender, its processus longer

than wide -> M. athalia; 3. uncus present, short and robus, its processus almost as long as wide

-> M. britomartis. In females there are slight differences among the species, but not all speci-

mens can be reliably identified in this way (Urbahn 1952; Batori et al. 201 2a).

Morphometric methods

Specimens used in this study were mounted and photographed on a millimeter grid from the

same angle (90°) using a DSLR camera (Canon 450D). Subsequently, 25 metric characters

were measured from the photographs using freeware ImageJ (Abramoff et al. 2004), 14 on

the forewing and 11 on the hindwing (Table 1, Fig. 1). The measurements included a wide

range of morphometric characters (i.e. lengths, surfaces and angles between measured dis-

tances of different parts of both wings). Moreover, one categorical character, i.e. marginal

line colour in contrast to lunular colour (for categories see Table 1 and Appendix A) was

also recorded. In order to exclude the impact of the animal’s size, all metric characters were

transformed into 18 additional ratios (Table 1). The photographs of both wings were used for

comparison and investigated in detail in order to find some other descriptive morphological

species-specific characteristics and to describe intraspecific variability. Although no body

asymmetry was detected, all characters were measured on the right side of an animal in order

to exclude any possible influence of this phenomenon. The left side, however, was used in

rare occasions when the right side was damaged.

Statistical analysis

Since in our samples males were prevalent and not all females could be reliably identified to

a species, females were excluded from subsequent statistical analyses. With this approach, we

JUGOVIC & KOREN: Wing pattern morphology of three closely related Melitaea...

Table 1. List of structures (with morphological characters, their explanation with measuring units) mea-

sured in three species of Melitaea.

No. | Character abbreviation Description Unit

Forewing

1 |FWS forewing surface mm?

2 |FWCS forewing cell surface mm?

3 (EWI forewing length 1 mm

4 |FWL2 forewing length 2 mm

5 |FWH forewing height mm

6 la angle at forewing apex 2

7 |Al distance Al mm

8 |A2 distance A2 mm

9 |A3 distance A3 mm

10 |A4 distance A4 mm

11 |FWDS forewing dot surface mm?

12 |FWLS forewing lunule surface mm?

13 |FWLL forewing lunule length mm

14 |FWLH forewing lunule height mm

Hindwing |

15 |AWS hindwing surface mm?

16 |HWCS hindwing cell surface mm?

17 |HWLI1 hindwing length 1 mm

18 | HWL2 hindwing length 2 mm

19 |HWL3 hindwing length 3 mm

20 |B angle at hindwing apex &

21 |Bl distance Bl mm

22 ıB2 distance B2 mm

23 |HWLS hindwing lunule surface mm?

24 |HWLL hindwing lunule length mm

25 IHWLH hindwing lunule height mm

Categorical character

(1=equal; 1.5=slightly darker; 2=darker; 3=much darker)

Ratios

26 > line colour in contrast to lunular colour

FWCS/FWS

FWL2/FWLI

FWH/FWLI

A2/Al

A3/Al

FWDS/FWCS

FWDS/FWS

34 \FWLH/FWLL

FWLS/FWS

FWLS/FWCS

B/a

HWCS/HWS

|HWLI/HWL2

_|HWLS/HWS

| HWLS/HWCS

|HWL3/HWLI

B2/B1

HWLH/HWLL

Nota Lepi. 37(1) 2014: 75-90 79

Figure 1. A photograph of Melitaea sp. with measured structures. Distances are denoted with full lines,

surface measurements with dashed lines and angles with Greek alphabet. Left: upperside; right: under-

side. For explanation of characters’ abbreviations see Table 1. For categorical character, see Appendix A.

also avoided the influence of possible sexual dimorphism. Only for the analysis of the marginal

line colour were a few unambiguously identified females added to the sample (no sexual dimor-

phism was noticed in this character). We also had to remove from the analysis three aberrant

individuals which lacked measuring points for some characters, hence they are commented on

separately.

For each species, the Kolmogorov-Smirnov test (at p = 0.05) and Normal Q-Q plots were

used to examine the normality of the data distribution, and the homogeneity of variances was

evaluated visually through scatterplots. The multivariate analysis of variance (MANOVA, at p

= 0.001) was used to test for significant differences between species. One-way analysis of vari-

ance (ANOVA) was used to assess the variation within a species for each character, significant

variation in a character being accepted if the between species variation was significant at p <

0.001. Pearson’s correlation coefficients (r) were computed to evaluate the extent to which each

character contributes unique information; only one character was chosen to represent a pair or a

group of characters where |r| > 0.9. The Durbin-Watson test (at p = 0.05 and p = 0.01) was ap-

plied to test for possible spatial (latitude, longitude, altitude) and temporal (year of collection)

autocorrelations of morphometric data (Savin & White 1977; Farebrother 1980) in each of the

three species. In subsequent multivariate statistical analyses, only selected characters (accord-

ing to the limitations listed above) were used.

80 JUGOVIC & KOREN: Wing pattern morphology of three closely related Melitaea...

Multivariate Principal Component Analysis (PCA) was used to identify the structure of our

data 1.e. to detect the possible influence of the species specific characteristics. Also, Discrimi-

nant Function Analysis (DFA) was carried out to examine possible separation of the three spe-

cies. In DFA, the contribution of each species was weighted according to its sample size (num-

ber of specimens). Post-hoc Games-Howell and Bonferonni tests were performed to assess

the rates of morphological divergences between pairs of species in details. The analyses were

performed using Microsoft Excell (2010), SPSS 14.0 for Windows (2005) (Norusis, 2005) and

Past (PAlaeontological STatistics) software (Hammer et al. 2001).

Results

The multivariate analysis of variance (MANOVA) showed significant differences between

species (p < 0.001). Out of 43 morphometric characters, 12 characters (without a single

ratio) were selected after Kolmogorov-Smirnov (p < 0.05), ANOVA (p < 0.001) and Pear-

son correlation (|r| < 0.9) tests. No spatial or temporal autocorrelation was detected in these

characters neither at p = 0.01 nor at p = 0.05 (Durbin-Watson test). Statistic description of

selected 12 characters (mean, standard error, 12th percentile, 88th percentile and extreme

values) is presented in Table 2. When the Pearson correlation test was set to |r| < 0.7, only

two morphometric characters (forewing height [F WH] and distance A4) were left after step-

wise exclusion of characters, meaning that correlations among most pairs of ne

characters were high.

The principal result of the PCA (Fig. 2) run on 95 males from the NW Balkans, using

12 morphometric characters (Table 2), revealed a significant overlap of the three species

along the first two principal components (PCs). PC 1 explained almost 70% of the total

variance and PC2 explained an additional 8%. All characters are positively correlated to

PCI, starting with the smallest M. aurelia and finishing with the largest of three species, M.

athalia. Nevertheless, the overlap between the three species in the middle section along PC

l is significant.

In the DFA (Fig. 3) run using the same morphometric characters as in the PCA, 95 males

were analyzed to (1) provide adequate species-grouping according to external morphologi-

cal characters and (2) detect intra-species variability of selected characters. Two Discriminant

Functions (DFs) explain total variance, DF 1 explaining over 75% of the total variance. Species

differ significantly only along DF 1 (p < 0.001), and only M. athalia differs significantly from

the other two species (Games-Howell Test, p < 0.001). The characters most correlated with DF

1 (indicated by the standardized discriminant function coefficients: DC > 0.75, Table 3) are:

forewing height (FWH), forewing lunule surface (FWLS), forewing lunule height (FWLH) and

hindwing surface (HWS). For the differences among the three species in these four characters,

see Fig. 4 and Table 2

Although the DFA aims to find the differences between the a priori defined groups, the mis-

classification rates were high, especially when cross-validation process was employed (Fig. 5).

Considering both DFs, only 67.4% specimens were correctly classified, and in the cross-valida-

tion procedure, the percentage dropped down to only 52.6% of correctly classified specimens.

We then repeated DFA with the same twelve morphometric characters, adding the coloration

of the marginal line in contrast to the lunular colour on the underside of the hindwing, not

Nota Lepi. 37(1) 2014: 75-90

Table 2. Statistical description of twelve metric characters in males of three species of the genus Melitaea

that were used in subsequent Principal component and Discriminant Function analyses. Upper row: aver-

age + std. error; lower row: (min), 12" percentile-88" percentile (max).

forewing height

forewing lunule | FWLS

surface

forewing lunule | FWLH

height

11.62 + 0.15 10.25 + 0.16 10.73 + 0.17

(9.27) 10.50-12.79 (13.56) (8.93) 9.15-11.46 (12.23) (9.19) 9.63-11.51 (12.78)

3.82 + 0.10 2.82 + 0.10 3.09 + 0.18

(2.01) 3.07-4.79 (4.98) (1.82) 1.95-3.55 (4.21) (1.74) 2.19-4.29 (5.28)

1.69 + 0.03 1.45 + 0.03 1.56 + 0.03

(1.21) 1.50-1.99 (2.09) (1.11) 1.24-1.61 (1.66) (1.33) 1.36-1.76 (1.79)

138.70 + 4.60 109.55 + 20.50 120.20 + 4.40

hindwing surface | HWS

(96.40) 106.64-171.47 (181.71)

(81.09) 89.78-144.80 (153.48)

(93.94) 103.09-161.06 (170.21)

se Bl 3.86 + 0.09 3.21+ 0.08 3.50 + 0.09

(2.76) 3.09-4.67 (5.10) (2.39) 2.63-3.76 (4.06) (2.63) 3.02-4.09 (4.57)

hindwing cell | HWCS 9.23 + 0.24 7.614026 7.94 + 0.26

surface (6.39) 7.35-11.42 (12.66) (5.17) 5.75-9.42 (10.85) (6.30) 6.60-9.44 (12.16)

Dr 1 8.53 + 0.14 7.54 +0.18 7.90 + 0.16

hindwing length 1

(6.48) 7.52-9.88 (10.15) (5.58) 6.45-8.78 (10.20) (6.44) 7.05-8.96 (9.40)

hindwing lunule | HWLS 4.29 + 0.15 3.39 + 0.14 3.58 + 0.20

surface (2.52) 3.14-5.32 (6.75) (2.08) 2.65-4.57 (5.00) (2.44) 2.53-4.82 (6.26)

hindwing lunule | HWLL 2.48 + 0.05 2.27 + 0.05 2.20 + 0.06

length (1.73) 2.02-2.85 (3.24) (1.68) 1.95-2.62 (2.79) (1.73) 1.81-2.49 (2.82)

hindwing lunule | HWLH 1.95 + 0.04 1.71 + 0.03 1.80 + 0.05

height (1.44) 1.69-2.34 (2.70) (1.28) 1.48-1.93 (2.16) (1.46) 1.55-2.08 (2.33)

. A4 1.84 + 0.05 1.59 + 0.05 1.59 + 0.05

distance A4

(1.09) 1.39-2.14 (2.49) (1.11) 1.32-1.92 (2.18) (1.13) 1.30-1.83 (2.01)

forewing lunule | FWLL 2.46 + 0.05 213.2005 2.19+ 0.09

length

(1.63) 1.98-2.80 (3.04)

(1.52) 1.83-2.40 (2.56)

(1.56) 1.59-2.75 (3.32)

0.0 1.6

PC 1 (69.8 %)

Figure 2. Two-dimensional plot (PC 1 vs. PC 2) generated from Principal Component Analysis (PCA) run

on 95 males of three Melitaea species using twelve metric characters (see Table 2). Legend: crosses: M

athalia; diamonds: M. britomartis; squares: M. aurelia.

82 JUGOVIC & KOREN: Wing pattern morphology of three closely related Melitaea...

+ M. athalia

M. aurelia

M. britomartis

M Group Centroid

ça

™

LL.

A 0 1

Function 1 (75.7 %)

Figure 3. Plot of the scores of 95 males of three species of Melitaea on both discriminant functions, using

12 morphometric characters. Centroids are larger and darker than symbols denoting individual specimens.

Table 3. Contributions of 12 metric characters (MC) to discrimination along both discriminant functions

(DF). Discriminant function analysis run on 95 males of three species of Melitaea. Asterisk (*) denotes

the highest absolute correlation between the character and DF.

MC DF 2

forewing height (FWH) 0.85 1* —0.002

forewing lunule surface (FWLS) 0.810* 0.110

forewing lunule height (FWLH) 0.791 —0.129

hindwing surface (HWS) 0.787* —0.022

distance B1 (B1) 0.680* —0.120

hindwing cell surface (HWCS) 0.651* 0.158

hindwing length 1 (HWLI) 0.624* —0.013

hindwing lunule surface (HWLS) 0.3793 0.128

forewing lunule length (FWLL) 0.371* 0.147

hindwing lunule height (HWLH) 0.562* -0.032

distance A4 (A4) 05337 0.285

hindwing lunule length (HWEL)

considering that this character was deviating from normal distribution (Kolmogorov-Smirnov

test, all data pooled: Z — 2.642; p < 0.001). However, this did not improve the classification

significantly (original grouping: 70.5%; cross-validation procedure: 55.8%).

In addition to the characters used in the DFA, two characters frequently mentioned in the

literature as diagnostic for the recognition of (some of) Melitaea species should be mentioned.

Nota Lepi. 37(1) 2014: 75-90 83

Lc]

FWLH (mm)

M. athalia M. aurelia M. britomartis M. athalia M. aurelia M. britomartis

Figure 4. Variation in four morphometric characters in M. athalia, M. aurelia and M. britomartis: fore-

wing height (FWH), forewing lunule surface (FWLS), forewing lunule height (FWLH) and hindwing

surface (HWS). Statistically significant differences between pairs of species (Bonferroni test) are shown

with capital (p < 0.001) and small (0.001 < p < 0.05) letters. When only a trend in differences between

species is shown (0.05 < p < 0.1), small letters with apostrophe are used.

For the characters like the spacing of the marginal, submarginal, postdiscal and discal line on

the forewing upperside, the colour of the submarginal spots on the hindwing underside, the

coloration of hairs on the palps as well as that of the marginal line in contrast to the lunular

colour on the underside of the hindwing, no consistency with species attribution was found

in our examination. For the latter, slight differences were found between the three species;

however, species could not be identified with certainty using the lunular coloration on the

hindwing underside due to the high intraspecific variability of this character. The coloration of

the marginal line is in general equal to the coloration of the lunules in M. athalia, much darker

in M. britomartis and only slightly darker in M. aurelia. Nevertheless, all of these categories

were noticed in each species (Fig. 6, Appendix B).

The wing pattern and coloration show high intra- and interspecies variability. While in some

species dark melanistic forms are common (e.g. in Melitaea britomartis ssp. michielii from

Slovenian Karst and surroundings (Carnelutti 1992)), other peculiar forms also had been found.

In some of these, the pattern deviated from normal form so much so that in these animals some

84 JUGOVIC & KOREN: Wing pattern morphology of three closely related Melitaea...

rad

©.

M. aurelia M. britomartis B M. athalia M. aurelia

Percentage

M. athalia M. aurelia M. britomartis M. athalia M. aurelia

Figure 5. DFA classification results of 95 males of Melitaea athalia (first column), M. aurelia (second

column) and M. britomartis (third column); A, B — original; C, D — cross-validation procedure.

EICH] MEERE

BER UNE ER

Percentage

M. athalia (N=66) M. aurelia (N=34) M. britomartis

(N=27)

Species

Figure 6. Colour categories of marginal line colour in contrast to lunular colour on underside of hindwing

of three Meliatea species (1=equal; 1.5=slightly darker; 2=darker; 3=much darker).

metric characters could not be measured. We found three such M. athalia specimens (i.e. with

extremely reduced markings) from Kamniski Vrh and Smrekovec (both in Slovenia) and Vu-

grovec (Croatia).

Nota Lepi. 37(1) 2014: 75-90 85

Discussion

The genitalia structures (Urbahn 1952) and the phylogeny based on three genes (mitochondrial

gene cytochrome oxidase subunit I, and two nuclear genes, elongation factor-la, and wingless;

Leneveu et al. 2009) of the three Melitaea species are in agreement, clearly showing that M.

aurelia is more distant from the other two species. In our analysis this is proved by a genitalia

based species identification of males without any questionable cases. However, the phylogenet-

ic relationships and the differences in the genitalic morphology are not reflected in the external

morphology of the three species’ wing patterns.

All analysis conducted in this study indicates the important differences among the three

species in the size of most of the measured characters, however, only in average values. Due

to high variability of these characters, the clinal variation among species has been noticed

(see the results of multivariate analyses). According to literature (e.g. Tolman & Lewington

2008), M. athalia is indeed the largest of the three species, with forewing height (FWH, Table

2, Fig. 4) being the most obvious character reflecting the difference in size from our study.

Without exception, all measured characters had the smallest average values in M. aurelia on

one side of the clinal variation and the largest in M. athalia on the other. High correlations

among most of the characters show the stability of their wing shape, which can therefore be

represented by a subset of all the characters. The wing shape stability is also supported by

the exclusion of all ratios (that partly exclude the size impact and describe the wing shape)

in ANOVA tests. In order to minimize the number of highly correlated characters, only one

character was chosen from a pair or group of highly correlated characters. This goal was

only partially achieved as further stepwise removal of characters (with Pearson correlation

coefficients 0.7 < |r| < 0.9) would result in only two weakly correlated remaining characters

(|r| < 0.7) and prevent the implementation of the multivariate analyses. Although the differ-

ences are small, M. athalia is the most distant from the other two in a powerful discriminant

function analysis. All except one character that were included in the multivariate statistical

analyses significantly separate M. athalia from the other two, and only slight differences in

just one (out of 43) metric character (forewing lunule height, FWLH; ANOVA: 0.05 < p <

0.1) between M. aurelia and M. britomartis were shown. Although the differences are small,

this further supports the discrepancies between the external morphology and phylogenetic

results (sensu Leneveu et al. 2009). Considering the phylogenetic position of the three spe-

cies, M. athalia should resemble M. britomartis more than M. aurelia but in our analysis

the latter two were clustered more closely together. According to the phylogenetic data, the

first split within the ‘athalia’ group that separated the clade containing M. aurelia from the

clade containing the two other species occurred in the Tortorian period (approximately 11

MY A) and the later split from which also M. athalia and M. britomartis emerged happened

during Messinian period (approximately 7.1 — 5.3 MYA; Leneveu et al. 2009). Hence, the

three lineages that emerged — M. athalia, M. aurelia and M. britomartis — underwent at least

5 million years of separate evolution. Despite such a long time of separation, no obvious ex-

ternal morphological differences have evolved. This is somewhat surprising when compared

to the situation in some of their close relatives. For example, clear differences in external

morphology have emerged in some species of Melitaea, although they have become separate

evolutionary lineages more recently than M. athalia, M. aurelia and M. britomartis (e.g. M.

86 JUGOVIC & KOREN: Wing pattern morphology of three closely related Melitaea...

asteria—M. aurelia, also within the same ‘athalia’ group; Tolman & Lewington 2008; Len-

eveu et al. 2009).

The allozyme polymorphism of these three species was studied in samples from the Car-

pathian basin, and revealed that M. britomartis and M. aurelia are more closely related, while

M. athalia appears to be a further relative (Batori et al. 2012a). This result contradicts the

phylogenetic analysis of Leneveu et al. (2009). The latter analysis, however, finds support

also in the genitalia structures as follows: in males of M. athalia and M. britomartis, the

spines on the uncus are well developed, whereas they are completely absent in M. aurelia

(Urbahn 1952). It should not be a surprise that the level of differentiation in genitalic struc-

tures corresponds to the phylogenetic position of the species since sexual differentiation

usually represents one of the most rapid and obvious taxon-specific events during speciation

in many animal groups, invertebrates in particular (Mayr & Ashlock 1991). In addition, the

genitalic structures are in comparison to the external morphology less subjected to envi-

ronmental factors (Cesaroni et al. 1994; Dapporto et al. 2011). For example, M. celadussa,

recently elevated to a species level by Leneveu et al. (2009) from a subspecies rank of M.

athalia, has not yet been reliably recognized on the basis of wing pattern and coloration,

whereas some differences from M. athalia were found in male genitalia alone (e.g. see Hig-

gins & Riley 1978; Leneveu et al. 2009). As long as external differences are not found, this

species pair could be treated as cryptic (sensu Hawksworth 2010: “populations which are

phylogenetically distinct, but distinguished by molecular or other features that are either not

evident macroscopically or generally overlooked’’). 3

In contrast to the allozyme study, the phylogenetic analysis used as a framework for the

explanation of our results (Leneveu et al. 2009) (1) included a vast majority of known species

of Melitaea (whereas in Batori et al. 2012a only M. athalia, M. aurelia and M. britomartis

were included) and (2) three different genes provided support for the same topology within the

‘athalia’ group discussed herein. Hence, no serious consideration was given to the study of

Batori et al. (2012a).

No external characters were proven to be reliable for species delimitation even though the

coloration of the marginal line of the hindwing underside shows the trend towards the correct

identification. Although the majority of specimens correspond to the character states given

in literature for a particular species, all three presumably species-specific categories of this

character (Tolman & Lewington 2008) used in our analysis were present in each of the three

species. Moreover, the percentage of misidentifications when this is the only character used

would be very high (see Fig. 6). Other external characters suggested in literature (see Intro-

duction) show even higher rates of variability, and no species-specific correspondence was

found. Even when the twelve (out of 43, see Tables 1 and 2) most powerful characters are

simultaneously used (see results of the DFA, Fig. 2) the misidentification rate reaches almost

50% in the cross-validation procedure (Fig. 5), rendering the appropriate identification of

these species highly unreliable. To further support the high morphological variability within

a single Melitaea species, as an example we should mention a comparison of populations

of M. athalia from the Caucasian basin (Batori et al. 2012b), where small, albeit signifi-

cant differences in wing morphology were found among some of them (all contributing to a

high intraspecies variability). Although partly reflecting the molecular differences between

Nota Lepi. 37(1) 2014: 75-90 87

the populations, these differences cannot be assigned to a genetic background alone. They

rather may through selection also be a result of an adaptation to local conditions (Batori et

al. 2012b), such as food availability, climate and microhabitat selection. This phenomenon

is not uncommon since high levels of phenotypic plasticity are important for a response

that ensures the survival of a population exposed to unstable environmental factors (Shapiro

1976; Brakefield & French 1999). High species plasticity together with accommodation to

local conditions can in practice result in description of many subspecies and morphological

forms within M. athalia as well as within some other Melitaea species. Nonetheless, the

differences among populations in reality might have resulted from locally specific selection

pressure (e.g. see Tolman & Lewington 2008). On the other hand, the three species exploit

similar (and partly overlapping) resources (e.g see Verovnik et al. 2012; Koren and Jugovic

2012), which is probably the reason for the high similarity among them. It is important to

note that the regional pattern of differentiation in M. athalia from the Caucasian basin was

less expressed in genitalia than in wing characters (Bätori et al. 2012b).

The high variability of these species is further shown in their qualitative characters, for

example in wing pattern and coloration. In M. athalia three out of more than 120 sampled

specimens had a very peculiar coloration, with no visible marginal lines. Albeit unrecognized,

we believe that differences in (some of) local conditions clearly show the importance of en-

vironmental factors for the wing morphology. This further demonstrates the importance of

genitalia or phylogeny based identification in the herein investigated Melitaea species. More-

over, the latter two approaches should have a significant advantage also for the identification

of (some) other species of the genus that should undergo thorough revision in the future (cf.

Leneveu et al. 2009).

Conclusions

The morphometric analysis of three closely related Melitaea species (M. athalia, M. aurelia

and M. britomartis) revealed the following: (1) small albeit significant differences in wing

morphology exist among the three species; (2) only characters describing the size of the spe-

cies with no characters describing their wing shapes (ratios) were statistically important for the

separation of the species; (3) whereas the structure of the male genitalia and the phylogenetic

position of the three species are concordant, the sequence of phylogenetic splits is not reflected

in the rate of external morphological differences among them; (4) our study represents another

example where external morphology based identification would lead to highly unreliable deter-

minations, hence a use of genitalia based identifications is strongly recommended.

Acknowledgements

We are grateful to Ivan Jugovic (Kranj, Slovenia) for data from his collection, useful discussion and

help in identification. We also thank Barbara ZakSek (Volicina, Slovenia) for providing us with the

literature on genitalia preparation, and Elena Varljen Buzan (Koper, Slovenia) for valuable comments

regarding the molecular background. We are thankful to Roman Luëtrik (Ljubljana, Slovenia) for valu-

able help with statistical analysis. The referees and the editor contributed greatly to the improvement

of the manuscript.

88 JUGOVIC & KOREN: Wing pattern morphology of three closely related Melitaea...

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Appendices

Appendix A. Three categories of marginal line colour in contrast to lunular colour: 1=equal (a); 2=darker

(b); 3=much darker (c) were used. In some cases when decision between categories 1 and 2 was hard (i.e.

when the two authors disagree on whether the category should be assigned to category 1 or 2), value 1.5

was used.

90 JUGOVIC & KOREN: Wing pattern morphology of three closely related Melitaea...

Appendix B. Representatives of three categories of marginal line colour in contrast to lunular colour

(categories 1, 2, 3) in Melitaea athalia (a-c), M. aurelia (d-f) and M. britomartis (g-1).

Nota Lepi. 37(1) 2014: 91-103 | DOI 10.3897/n1.37.7928

Choreutidae of Madeira: review of the known species and description

of the male of Anthophila threnodes (Walsingham, 1910) (Lepidoptera)

JADRANKA ROTA!, ANTONIO M. F. AGUIAR”, OLE KARSHOLT*

1 Laboratory of Genetics/Zoological Museum, Department of Biology, University of Turku, FI-20014 Turku,

Finland; jadranka.rota@utu.fi

2 Laboratorio de Qualidade Agricola, Entomologia, Caminho Municipal dos Caboucos, 61, 9135-372 Camacha,

Madeira, Portugal; antonioaguiar.sra@gov-madeira.pt

3 Zoological Museum, University of Copenhagen, Universitetsparken 15, DK-2100 Copenhagen, Denmark;

okarsholt@snm.ku.dk

http://zoobank.org/9CD3F560-D46D-4E63-A309-E74D061799E7

Received 13 March 2014; accepted 10 May 2014; published: 15 June 2014

Subject Editor: Erik van Nieukerken

Abstract. We review and illustrate the four species of Choreutidae recorded from Madeira — Anthophila

threnodes (Walsingham), A. fabriciana (Linnaeus), Choreutis nemorana (Hübner), and Tebenna micalis

(Mann) — and describe and illustrate for the first time the male of A. threnodes, as well as the biology of

this Madeiran endemic. We provide brief notes on each of the species and give short diagnoses for cor-

rectly identifying them. Finally, we discuss previous misidentifications of Madeiran choreutids and the

occurrence of choreutids on other oceanic islands.

Introduction

The Lepidoptera fauna of the Madeira Islands consists of only 331 species (Aguiar & Karsholt

2008). This is mainly due to the isolated position of these islands in the Atlantic Ocean, and

only to a lesser extent to insufficient collecting efforts. The Macrolepidoptera fauna, and es-

pecially the butterflies (Papilionoidea), are considered to be well known, with only a few and

mostly invasive species being added in recent years. Among the Microlepidoptera, new discov-

eries still occur regularly, and a number of taxonomic problems still await resolution.

Madeiran lepidopterology began with Thomas Vernon Wollaston who, for health reasons,

stayed in Madeira for long periods between 1847 and 1855 and also made later visits to the

island. His results, including descriptions of many new species, were published by himself

(Wollaston 1858) and Stainton (1859). Subsequent important contributions to the Lepidoptera

fauna of the Madeira Islands were made by Walsingham (1894) and Rebel (1917, 1940). Eas-

ier access to the islands by airplane and the use of modern equipment (e.g., mercury vapour

lamps) during the second half of the 20" century resulted in an increasing number of specimens

available for study. Results of these efforts were summarized in the recent catalogue by Aguiar

& Karsholt (2006), and data from that work were used for the checklists by Aguiar & Karsholt

(2008) and Karsholt & Nieukerken (2011). A brief introduction to lepidopterology in Madeira

was published by Karsholt (2000).

92 Rota et al: Choreutidae of Madeira: review of the known species...

Below we treat the small family Choreutidae. Currently, 413 species of choreutids are

described, and most of them are found in the tropics (Rota unpublished database). They are

usually diurnal and often brightly coloured. So far four species have been found in Madeira.

We describe and illustrate the unknown male and the biology of the endemic Anthophila

threnodes (Walsingham), and provide images and brief notes on the other three species: An-

thophila fabriciana (Linnaeus), Choreutis nemorana (Hübner), and Tebenna micalis (Mann).

At the end we discuss earlier misidentifications of Madeiran choreutids.

Methods

Genitalia dissections and terminology follow Rota (2008a). Plant names are from the Plant

List website (2010). Photographs of adults were taken using Leica Application Suit MZ 16A

and Zerene Helicon Stacker program for stacking subsequent images at ZMUC. Photographs

of genitalia slides were taken using an Olympus SZX16 microscope with motorized focus

drive attached to an Olympus E520 digital camera and they were then combined by using

the programs Deep Focus 3.1 and Quick Photo Camera 2.3 at the Zoological Museum of the

University of Turku. All images were improved in Adobe Photoshop CS3.

We conducted standard DNA extraction from abdomens of two specimens of Anthophila

threnodes (voucher codes noted below) and attempted standard PCR amplification of the DNA

barcode (cytochrome oxidase subunit I) using primers LCO and HCO (Folmer et al. 1994).

When this failed, we used a primer internal to the DNA barcode (K699; Mitchell et al. 2005)

in combination with LCO and successfully amplified 326 base pairs from one of the specimens

(DNA voucher An_th2; see details under material examined). As this fragment was very short,

we attempted to obtain more sequence data and after some trials with the commonly amplified

nuclear genes in Lepidoptera (Wahlberg and Wheat 2008), we were finally successful in ob-

taining two fragments of the nuclear gene GAPDH using newly designed primers by Niklas

Wahlberg (GAPIF/GAP2R and GAP3F/GAPAR; primer sequences in Table 1), resulting in a

total of 580 base pairs of GAPDH. We carried out a phylogenetic analysis of the concatenated

sequences from À. threnodes together with the sequences of four other species of Anthophila,

and another choreutid, Prochoreutis inflatella, as an outgroup (Appendix), using MrBayes v.3.2

(Ronquist et al. 2012) running on the CIPRES server (Miller et al. 2010). The data were ana-

lysed unpartitioned, with two concurrent runs each with one cold and three heated chains for

one million generations, and 25% of trees were discarded as burnin. Convergence was assessed

by ensuring that standard deviation of split frequencies was well below 0.05 (it was 0.001 at the

end of the analysis), that PSRF values were all very close to 1.000 (they ranged 1.000-1.005),

and by visual inspection of plots of log likelihood and all parameters in Tracer v.1.5 (Rambaut

and Drummond 2007). Sequence length and GenBank accession number for each species are

listed in the Appendix.

Data from the type material are cited literally whereas data from the other material are given

in a standardized format.

Nota Lepi. 37(1) 2014: 91-103 93

Table 1. Primers used for PCR amplification of the nuclear gene GAPDH.

Primer pairs and sequence

GAP1F (AARGCTGGRGCTGAATATGT) / GAP2R (TAACTTTGCCRACAGCYTT)

GAP3F (GTGCCCARCARAACATCAT) / GAP4R (CGGCTGGAGTARCCATAY TC)

Abbreviations

AMFA Collection of Antonio M. F. Aguiar, Funchal, Madeira.

BMNH The Natural History Museum, London, U.K.

ICLAM Insect Collection Laboratorio de Qualidade Agricola, Madeira, Portugal

SIP Collection of Leo Sippola, Pirkkala, Finland

ZMUC Zoological Museum, University of Copenhagen, Denmark

Anthophila threnodes (Walsingham, 1910)

http://species-id.net/wiki/Anthophila_threnodes

Figs 1, 2, 9-18

Hemerophila threnodes Walsingham, 1910: 257.

Material examined. Holotype. 19, ‘B.M. Genitalia Slide 20115 A. Diak’ | ‘Type’ "Madeira, V. 1886, Leech, 62296’ |

(1/1) descr. 62296’ (BMNH). 14, 19, ‘Madeira: 1858. Wollaston. BM 1858-21.’ | ‘Simaethis fabriciana L. teste Stn.

Ann-Mag. N. H. (3. s) III. 210, no 4 (1859), Topotype Madeira.’ | ‘V’ [May] (BMNH); 14, Funchal, 20-31 .iii.1995,

leg. L. Sippola, genitalia slide Rota JR2013-05, DNA voucher for extraction An th2 (ZMUC); 14, Cabo Giräo, Cruz

da Caldeira, 8.vii.1999, leg. A. M. F. Aguiar, genitalia slide Rota JR2013-04, DNA voucher for exraction An th]

(AMFA); 19, Vereda da Entrosa, Arco de Sao Jorge, 225 m, 12.11.2000, leg. A. M. F. Aguiar (AMFA); 34, 29, same

data but la. 14.11.2001, Urtica sp., leg. A. M. F. Aguiar & J. Jesus (AMFA, ICLAM); 25, 39, same data but 200 m, la.

29.x1.2001, Urtica sp., leg. A. M. F. Aguiar & O. Karsholt, genitalia slides Hendriksen 5240, Karsholt 5236 (AMFA,

ZMUC); 29, same data but, 15.v.2003, leg. A. M. F. Aguiar & J. Jesus (AMFA); 19, AM 5089, Levada Grande, Boa-

ventura, 285m, 4.ii1.2000, larva on Urtica membranacea, leg. A. M. F. Aguiar (AMFA); 19, Monte, 20.iii.2002, leg.

L. Sippola (SIP).

Remarks. Hemerophila threnodes was originally described from one female collected in Ma-

deira by J. H. Leech in May 1886. No exact locality was stated. Based on the material collected

more recently, we provide a description of the male. We also illustrate female genitalia.

Description. 4. threnodes is characterized by its dark, almost black wings and a scattering

of light bluish scales on the thoracic dorsum and forewing upperside (Figs 1, 2). The forewings

have a cream-white spot at two-thirds of costa and another such spot at four-fifths of the dorsum;

fringes are black and cream-white beyond black fringe-line. Hindwings are uniform dark brown.

Male genitalia (Fig. 9). Tegumen triangular with small uncus extending from its apex. Pa-

pillae anales present as elongate, somewhat elliptical patches with long hairs. Gnathos well

developed as a long, somewhat curved, pointed hook. Vinculum ventrally rounded with a small,

triangular saccus (obscured in Fig. 9a, but visible in slide JR2013-04, which is not illustrated).

Valva broad, somewhat oval, with a pointed costal process and an unsclerotized triangular,

rounded extension distally; distally and ventrally covered with hairs. Juxta as a hood-like plate

(in Fig. 9b attached to phallus). Phallus shorter than valva, slightly sigmoidal, with a sharp

spine at one-third from apex (Fig. 9c).

94 Rota ef al: Choreutidae of Madeira: review of the known species...

; x i u

Figures 1-4. Adults in dorsal and lateral view. 1-2. Anthophila threnodes; 3-4. Anthophila fabriciana (scale

bar — 2 mm).

Female genitalia (Fig. 10). Apophysis posterioris slender, slightly broader at base; ca. 1.5

times as long as anterioris and much less thick. Apophysis anterioris greatly enlarged in the

middle, tapering basally and even more so distally. Ostium on segment 7. Ductus bursae very

gradually widening into corpus, with a slight twist of about one to two revolutions. Corpus

bursae oval, small, with a signum as small patch of dentations (Fig. 10b).

Host plant. Urtica membranacea Poir. ex Savigny and probably other Urtica spp. (Urti-

caceae).

Remarks. Larvae have been found in March, May, and November, and adults have been col-

lected in February, March, May, and July, indicating at least two broods. The adult flies during

the day. It occurs in open landscapes at low altitudes.

Immature stages and biology (Figs 11-18). Larva is off-white with dark brown spots (Figs

14, 15) and it spins a thin web on or around the young leaves on which it feeds (Figs 12, 13).

The pupa is reddish brown (Fig. 16), and in addition to the single rows of dorsal spines on

abdominal segments A2-7 (Figs 17, 18) it also has dorsal lacunae — small round holes in a row

immediately posterior to the spines.

Diagnosis. A. threnodes is characterized by its blackish wings, and should be relatively easy

to differentiate from the other Lepidoptera in Madeira. It resembles A. fabriciana but the wings

of that species are much lighter greyish brown. Male genitalia are very similar to those of A.

fabriciana, but they differ in having a much shorter spine on the phallus. Female genitalia are

similar to those of other species of Anthophila, but they differ in only having a hint of spiraliza-

Nota Lepi. 37(1) 2014: 91-103 95

tion in the basal third of the ductus bursae, unlike those of e.g. A. fabriciana (L.), A. abhasica

Danilevsky, and A. armata Danilevsky, where the spiralization is apparent. We cannot find dif-

ferences between the larvae of A. threnodes and A. fabriciana in their general appearance (the

differentiation between the two would likely be possible based on the ultrastructure as seen with

the scanning electron microscope, but this has not been done).

Anthophila fabriciana (Linnaeus, 1767)

Figs 3, 4

Phalaena (Tortrix) fabriciana Linnaeus, 1767: 880

Material examined. | 9, Vereda da Entrosa, Arco de Sao Jorge, 225 m, 14.11.2001, leg. A. M. F. Aguiar (ICLAM).

Host plant. Urtica sp. (Urticaceae).

Remarks. This is a new record for Madeira. The single specimen was collected at the same

place as several A. threnodes. We examined photographs of this specimen deposited in the

BMNH and are of the opinion that it is correctly identified.

Diagnosis. A. fabriciana resembles A. threnodes, but differs by its greyish brown wings. It

has the cream-white spots apically at costa and dorsum connected by a light zigzag-line, and

has a white streak near the margin in the hindwings (Figs 3, 4).

Choreutis nemorana (Hübner, 1899)

http://species-id.net/wiki/Choreutis_nemorana

Figs 5, 6, 19, 20

Tortrix nemorana Hübner [1799]: pl. 1, fig. 3.

Material examined. Serra d’ Agua, Pousada dos Vinhaticos, 660 m, 104, 139, 12-13.ix.1975, leg. O. Lomholdt &

N. L. Wolff; Säo Vicente, sea level, 19, 16.vi.1993; 54, 39, same data, but larva on Ficus carica, leg. O. Karsholt

(ZMUC); Sao Vicente, Ribeira do Inferno, 79m, 3, 12.ix.1996, leg. F. Aguiar & J. Jesus (AMFA, ICLAM); Ponta do

Pargo, Porto do Pesqueiro, 311 m, 14, 19, 23.iv.1994, leg. A. M. F. Aguiar (AMFA); Fajä da Nogueira, 600-1000 m,

19, 8.x.1994, leg. O. Karsholt (ZMUC); Curral das Freiras, 850 m, 14, 20.ix.1997, leg. O. Karsholt (ZMUC); same

data, but 597 m, 34, 29, la. 16.iv.1998, Ficus carica, leg. F. Aguiar & J. Jesus; 34, 19, same data, but 7.v.1998

(ICLAM); Estreito de Camara de Lobos, Levada do Norte, Garachico, 14, 19, 538 m, 26.vii.2001, J. Jesus (AMFA,

ICLAM).

Host plant. Ficus carica L. (Moraceae).

Remarks. This species is common wherever there are Ficus carica trees. Larvae, hiding

under a thin web, skeletonise mainly young leaves (Figs 19, 20). They have been found in June,

and adults have been collected in April-July and September-November, at altitudes from sea

level to 1000 m. C. nemorana is only a minor pest on figs in Madeira. The adult flies during

day. Male and female genitalia were illustrated by Diakonoff (1986): pl. 66, fig. 81 (male), and

pl. 142, fig. 81 (female).

Diagnosis. The almost square, brown forewings, and the black and yellow hindwings make

C. nemorana unmistakeable among Madeiran Lepidoptera (Figs 5, 6).

96 Rota ef al: Choreutidae of Madeira: review of the known species...

Figures 5-8. Adults in dorsal and lateral view. 5-6. Choreutis nemorana; 7-8. Tebenna micalis (scale bar

= 2 mm).

Tebenna micalis (Mann, 1857)

http://species-id.net/wiki/Tebenna_micalis

Figs 7, 8

Choreutis micalis Mann, 1857: 181.

Material examined. Funchal, Lido, 14, 20.viii.1974, leg. E. Traugott-Olsen, genitalia slide Wolff 4298; same

E. Traugott-Olsen, genitalia slide Wolff 4294, Rasmussen 4849; same locality, but 600-1000 m, 24, 8.x.1994,

leg. O. Karsholt, genitalia slide Hendriksen 5765; same locality, but 700 m, 34, 29, 18.ix.1997, leg. O. Karsholt;

Wolff (ZMUC); Ponta de Säo Lourenco, sea level, 94, 49, 24-27.vi.1993, 19, 11.vii.1993, 14, 17.ix.1997, leg.

O. Karsholt (ZMUC); Machico, sea level, 14, 27.vi.1993, leg. O. Karsholt (ZMUC); Achadas da Cruz, 725 m, 36,

2°, 8.vii.1993, leg. O. Karsholt (ZMUC); Porto Moniz, sea level, 19, 9.x.1994, leg. O. Karsholt (ZMUC); Santo

5766 (ZMUC); 14, Estreito de Camara de Lobos, Jardim da Serra, 1130 m, 22.x.1998, leg. F. Aguiar & J. Jesus

(ICLAM); 1°, Porto da Cruz, Chao das Feiteiras, 1251 m, 12.xi.1998, leg. A. M. F. Aguiar (AMFA); 16, 59, Santa-

na, Pico, Posto Agrario, 411 m, la. 12.x1.1998, Arctium minus; same data, but 19 1.vi.1999, leg. F. Aguiar & J. Jesus

(ICLAM); 17, Boaventura, Vereda da Entrosa, 130 m, 12.11.2000; 54, 39, same data, but 175 m, la. 14.ii.2001,

Nota Lepi. 37(1) 2014: 91-103 97

Figures 9-10. Anthophila threnodes: male genitalia (9a), phallus (9b, c) (9a and 9b from slide Karsholt 5236,

ZMUC; 9c from slide JR2013-04, ZMUC), female genitalia (10a), inset showing magnified corpus bursae

(10b) (scale bar = 0.2 mm).

Helichrysum melaleucum, leg. F. Aguiar & J. Jesus (AMFA, ICLAM); 14, 14, Estreito de Camara de Lobos, Levada

do Norte, Garachico, 538 m, 26.vii.2001 leg. J. Jesus (ICLAM); 19, Machico, Funduras, 605 m, 12.vi.2003, leg. J.

Jesus (ICLAM); 19, Santana, Achada do Gramacho/ Cais de Sao Jorge, 267 m, 21.vii.2011, leg. J. Jesus (CLAM);

14, Camacha, Levada dos Tornos, direction Camacha — Monte, 788 m, Cirsium vulgare, la. 13.vi.2013, leg. S.

Fontinha (ICLAM).

Host plants. Arctium minus (Hill.) Bernh., Cirsium vulgare (Savi) Ten., Helichrysum foeti-

dum (L.) Cass. and Helichrysum melaleucum Rchb. (Compositae).

Remarks. Larvae have been found in October, November, February, June, and July, and

adults have been collected in all months from June to November, at altitudes from sea level

to 1250 m. The adult flies during the day and comes to light. This is a widespread species. In

addition to being present on Madeira, it occurs on the Canaries, throughout the Mediterranean,

from central and eastern Europe throughout southern Asia to China and Japan, as well as pos-

sibly on Marianne Islands, Java, and New Zealand (Diakonoff 1986). It is the only choreutid

occurring in the Azores Islands, where it inhabits all larger islands (Karsholt & Vieira 2005).

98 Rota ef al: Choreutidae of Madeira: review of the known species...

Figures 11-16. Anthophila threnodes: 11. Adult on its host plant; 12-13. Larval webbing tying young

leaves; 14-15. Larva; 16. Pupa and an empty pupal shell in the inset.

Male and female genitalia were illustrated by Diakonoff (1986): pls. 46, 47, figs 54-1 and 54-2

(male), pl. 127, fig. 54 (female).

Diagnosis. With its sub-triangular forewings with black and metallic markings adult T

micalis is unique among Madeiran Lepidoptera (Figs 7, 8). It can be separated from the sim-

ilar T. bjerkandrella (Thunberg, 1784) by the presence of a subplical black and metallic spot

in the forewing in 7: micalis; such a spot is missing in 7: bjerkandrella (Diakonoff 1986).

Molecular results. The successfully amplified A. threnodes COI haplotype is unique and

it differs from the A. alpinella haplotype in 14 bases, and from the A. fabriciana haplotype in

12 bases. In the resulting phylogenetic tree, Anthophila threnodes and A. fabriciana are sister

Nota Lepi. 37(1) 2014: 91-103 99

Figures 19-20. Choreutis nemorana: larva under its webbing on the host plant Ficus carica (19) and larva

exiting its web-shelter after being disturbed (20).

species, but without statistical support (PP or posterior probability = 0.81). Together, they are

strongly supported as being the sister group to the North American species Anthophila alpinella

(PP=1).

Discussion

One of the aims of this paper was to provide the necessary information for correctly identifying

Madeiran choreutids because misidentifications have been common in the past. For example,

A. threnodes was misidentified as A. fabriciana by Stainton (1859) (as ‘Simaéthis Fabriciana

Linnaeus’) and subsequent authors, and it remained as such on the list of Madeiran Lepidoptera

100 Rota ef al: Choreutidae of Madeira: review of the known species...

Prochoreutis inflatella

Anthophila sp. (Rwanda)

Anthophila sp. (Peru)

Anthophila alpinella

Anthophila fabriciana

Anthophila threnodes

Figure 21. Bayesian tree showing the relationships among species of Anthophila. The numbers below

branches are posterior probabilities.

until 2006 (Aguiar & Karsholt 2006). The specimen of A. fabriciana listed above is the only

specimen of that species known from Madeira, while all the other ones previously identified

as A. fabriciana are actually A. threnodes. Although A. fabriciana is a common species in

mainland Europe, in addition to being recorded only once from Madeira, it has also been found

only once in the Canary Islands, “based on a single specimen (“61978”), taken in April 1884

[in Tenerife], by the late Mr. J. H. Leech” (Walsingham 1908). The Lepidoptera fauna of the

Canary Islands (and especially Tenerife) is relatively well studied, and it is surprising that A. fa-

briciana had not been found again in the islands, suggesting that the species failed to establish

itself there, which is in agreement with Rebel’s opinion (1911) that the single specimen might

have resulted from an accidental importation.

Furthermore, Tebenna bjerkandrella (Thunberg), a species similar to T. micalis, has been

noted as found in Madeira several times (e.g., by Walsingham 1894, 1908; Rebel 1911). How-

ever, all Madeiran specimens examined by us belong to the latter species, and the presence of

bjerkandrella in Madeira requires confirmation. It is likely that records of bjerkandrella from

other Macaronesian archipelagos also refer to T. micalis (Aguiar & Karsholt 2006).

The biology of the immature stages of A. threnodes is typical for the whole family. Spinning

a thin web on or around the young leaves on which the larva feeds is known from a number

of other genera (Rota 2005, Rota 2008b). Likewise, the morphology of the pupa is also shared

with several other genera. For example, the dorsal lacunae (Figs 17, 18) that are found in the

pupa of A. threnodes are also known to occur in the other species of Anthophila (Patocka 1999),

as well as in Asterivora Dugdale (Dugdale 1979), Caloreas Heppner (Keifer 1937), Hemero-

phila Hübner (Rota unpublished), Rhobonda Walker (Rota 2005), Prochoreutis Heppner, and

Tebenna Billberg (Patoëka 1999). Their function remains unknown.

Finally, we discuss other choreutids that are known to occur on oceanic islands. For exam-

ple, Brenthia leptocosma Meyrick is currently known only from Mauritius (Williams 1951).

Then there is a species of Choreutis that appears to be a pest on Ficus sp. (Moraceae) on the Ha-

walian islands (W. Nagamine, pers. comm.), and another species, Niveas kone Rota, is known

from the Solomon Islands and Papua New Guinea (Rota and Miller 2013). All of this suggests

that choreutids disperse fairly well, but most likely through passive wind dispersal because they

are small moths without strong flight capabilities.

Nota Lepi. 37(1) 2014: 91-103 101

As À. threnodes is endemic to Madeira, it would be interesting to firmly establish its sister

species so as to try to ascertain the origin of this species. We attempted to do so using molec-

ular data, but were highly limited by the small amount of sequence that we obtained from our

material and therefore our finding that A. fabriciana is the sister species of A. threnodes can

only be taken as a preliminary result although this also appears to be supported by the close

similarity of the two species in their external appearance, as well as their genitalia. Ideally,

molecular work employing phylogeographic methods, conducted on freshly collected material

of a large number of specimens of A. alpinella, A. fabriciana, and A. threnodes, as well as other

potential close relatives of A. threnodes, would provide an answer with more certainty than we

can do at this point.

Acknowledgements

We are grateful to Kevin Tuck, The Natural History Museum, London, U.K. for access to the collections

and for information and photographs of specimens; to Jurate De Prins, Royal Museum for Central Africa,

Belgium; and Leo Sippola, Pirkkala, Finland for loan of specimens. We greatly appreciate the input on an

earlier version of the manuscript provided by Martin Corley and Marko Mutanen. JR was funded by the

Finnish Kone Foundation while doing this research. OK’s work on the Lepidoptera of Madeira project

was supported by a grant from the Carlsberg Foundation (Denmark).

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103

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Nota Lepi. 37(1) 2014: 105-106 | DOI 10.3897/n1.37.7955

Book review: I Lepidotteri del Parco Naturale delle Capanne di

Marcarolo

OLE KARSHOLT !

1 The Natural History Museum of Denmark, Copenhagen, Denmark; okarsholt@snm.ku.dk

http://zoobank.org/5 9A SA3BD-242F-4749-85C5-9376144BDB70

Received 11 February 2014; accepted 12 February 2014; published: 15 June 2014

Subject Editor: Jadranka Rota

Giorgio Baldizzone, Carlo Cabella, Faustino Fiori & Pier Giuseppe Varalda, 2013. I Lep-

idotteri del Parco Naturale delle Capanne di Marcarolo. Memorie dell’ Associazione Nat-

uralistica Piemontese, vol. 12: 1-316, pls 1-32. ISBN: 978-88-902859-5-0

Can be obtained from the first author (giorgiobaldizzone@tin.it). Price: see below.

The nature park Capanne di Marcarolo is situated in north-western Italy north-west of Genova

on the border between Piedmont and Liguria, viz. in the transition zone between the Alps and

the Apennines. It is a so-called ‘regional park’ and has an area of about 82 km’. It ranges from

335 m to 1772 m (Monte delle Figne) in altitude and includes a number of vegetation types.

Much of the original forest has been cut in former times and partially replaced by pines. The

park has different types of wetlands including micro-peatbogs. One can get a good impression

about the park from a video on YouTube (http://www. youtube.com/watch?v=uPKTKO131HU).

This publication presents the result of the study of the Lepidoptera fauna of Capanne di

Marcarolo during the years 2002 to 2012, and it is the first book that treats all species of Le-

pidoptera (micro and macro) of a protected area in Italy. The research has served to draw up an

action plan for the management and conservation of the Lepidoptera of the protected area and

has resulted in the approval of the plan by the regional government of Piedmont. It is written in

Italian, but as the main part of the book is a systematic review of the collected specimens the

language should for most foreign readers of the book not be a serious problem (for writing this

review I had good help from Google translate).

The book starts with a summary (also in English), and continues with a preface, an intro-

duction to the study area (3 pp.), climate (5 pp.) and flora and vegetation (3 pp.) (these three

chapters are written by specialists other than the authors). Then follows a general chapter on the

entomological fauna of Capanne di Marcarolo (6 pp.) and a chapter with acknowledgements.

The main part of the book (pp. 34-284) presents the results of the study. Each of the 1464

(sic!) recorded species is presented with full name (with author and year of description). The

studied material with exact dates and localities is listed. For each species both the general distri-

bution and the distribution in Italy are summarized, with references to literature, as well as host

106 KARSHOLT: Book Review - I Lepidotteri del Parco Naturale delle Capanne di Marcarolo

plants and other biological details. For a few species there are more detailed remarks (like the

description of the hitherto unknown female genitalia of Homoeosoma incognitellum (Roesler)).

Even though a lot of progress has be made in the study of the Lepidoptera fauna of Mediter-

ranean countries during the last decades our knowledge of some families of Microlepidoptera

is still imperfect, and hardly any person is able to recognize all species of Lepidoptera occur-

ring in an area like Capanne di Marcarolo. The authors have solved this problem by allying

themselves with a number of taxonomic specialists of different families of Microlepidoptera.

Thereby (probably) all collected specimens have been identified probably as well as possible

at the present time.

The systematic part is followed by chapters with a discussion and conclusion (4 pp.), an ex-

tensive reference list (8 pp.) and indexes to Lepidoptera and plant names. The book is complet-

ed by a map of the area and 32 colour plates with photographs of aspects of climate, geological

features, plant species, characteristic biotopes and, especially, selected species of Lepidoptera

occurring in the studied area.

The book is published as a volume of Memorie dell’Associazione Naturalistica Piemontese

and founded by Fondazione Cassa di Risparmio di Torino. It is not literally for sale, but can

be obtained from its first author by giving a donation of at least 25€ plus shipping costs. The

donation will be used for research and publications dedicated to the knowledge of the Lepidop-

tera fauna of Italia habitats included in the Natura 2000 network. This is an interesting kind of

crowdsourcing, and both the idea and the book are herewith warmly recommended.

SOCIETAS EUROPAEA LEPIDOPTEROLOGICA e.V.

Nota Lepidopterologica wird als wissenschaftliche Zeitschrift von der Societas Europaea Lepidopterologica

(SEL) herausgegeben und den Mitgliedern der SEL zugesandt. Autoren, die Manuskripte fiir die Publikation

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adhésions ou au Trésorier.

SMITHSONIAN INSTITU

ALL TE

3 9088 0175

Contents mme

Nota Lepidopterologica combines tradition and innovation through open access and advanced

publishing model

Jadranka Rota

The first record of Pyrgus malvoides (Elwes & Edwards, 1897) in the Czech Republic

(Lepidoptera, Hesperiidae): an accidental introduction?

Vladimir Hula, Jana Niedobova

Revision of the Palaearctic and Oriental species of the genus Naarda Walker, 1866

(Lepidoptera, Erebidae, Hypeninae). Part 3. Description of three new species from Asia

Balazs Toth, Laszlé Ronkay

Reproductive isolation and intraspecific structure in Alpine populations of Erebia euryale

(Esper, 1805) (Lepidoptera, Nymphalidae, Satyrinae)

Frans Cupedo

Scotopteryx kurmanjiana, a new species from the Kopet-Dagh Mountains (Lepidoptera,

Geometridae, Larentiinae)

Hossein Rajaei Sh., Gyula M. Läszlô

Description of the reduced mouth parts of Coleophora micronotella Toll (Lepidoptera,

Coleophoridae), with a new synonym

Bernard Landry, Giorgio Baldizzone

On the taxonomic status of Ochromolopis ictella (Hübner, 1813) and O. zagulajevi Budashkin

& Sachkov, 1991 (Lepidoptera, Epermeniidae)

Reinhard Gaedike, Richard Mally

Description of the female of Ethmia cribravia Wang and Li 2004 (Lepidoptera, Elachistidae,

Ethmiinae)

Dmitry E Shovkoon, Tatiana A. Trofimova

A remarkable new species of the genus Catatinagma Rebel, 1903 (Lepidoptera, Gelechiidae)

from Turkmenistan

Oleksiy V. Bidzilya

Wing pattern morphology of three closely related Melitaea (Lepidoptera, Nymphalidae)

species reveals highly inaccurate external morphology-based species identification

Jure Jugovic, Toni Koren

Choreutidae of Madeira: review of the known species and description of the male of

Anthophila threnodes (Walsingham, 1910) (Lepidoptera)

Jadranka Rota, Antonio M. E Aguiar, Ole Karsholt

Book review: I Lepidotteri del Parco Naturale delle Capanne di Marcarolo

Ole Karsholt

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