Corresponding author: Stanislav K. Korb ( firstname.lastname@example.org )
Academic editor: Thomas Schmitt
© 2016 Stanislav K. Korb, Zdenek F. Fric, Alena Bartoňová.
This is an open access article distributed under the terms of the Creative Commons Attribution License (CC BY 4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
Citation: Korb SK, Fric ZF, Bartoňová A (2016) Phylogeography of Koramius charltonius (Gray, 1853) (Lepidoptera: Papilionidae): a case of too many poorly circumscribed subspecies. Nota Lepidopterologica 39(2): 169-191. https://doi.org/10.3897/nl.39.7682
Koramius charltonius (Gray, 1853) (Lepidoptera: Papilionidae) is distributed in the mountains of Central Asia. We analysed genetic and phylogeographic patterns throughout the western part of its range using a mitochondrial marker (COI). We also analysed the wing pattern using multivariate statistics. We found that the species contains several unique haplotypes in the west and shared haplotypes in the east. The haplotype groups do not correspond to the wing pattern and also the described subspecies do not correspond to either the haplotypes or the groups circumscribed by the wing pattern. Currently, there are more than ten subspecies of K. charltonius in Central Asia; based on our analyses we suggest a reduction to only five of them. The following nomenclatural changes are applied: (1) K. charltonius aenigma Dubatolov & Milko, 2003, syn. n., K. charltonius sochivkoi Churkin, 2009, syn.n., and K. charltonius alrashid Churkin & Pletnev, 2012, syn. n. are new synonyms of K. charltonius romanovi (Grum-Grshimailo, 1885); (2) K. charltonius marusya Churkin & Pletnev, 2012, syn. n., K. charltonius eugenia Churkin, 2009, syn. n., K. charltonius anjuta Stshetkin & Kaabak, 1985, syn. n., and K. charltonius mistericus Kaabak, Sotchivko & Titov, 1996, syn. n. are new synonyms of K. charltonius vaporosus (Avinov, 1913); and (3) K. charltonius safronovi Korb, Shaposhnikov, Zatakovoy & Nikolaev, 2013, syn. n. is a new synonym of K. charltonius voigti (Bang-Haas, 1927).
The systematics of Parnassiinae (Papilionidae, Lepidoptera) is complicated and has been a subject of many debates. The first classification was created by
Molecular studies in the last two decades did not change these systems in general.
Using all the available data on Parnassiinae and published phylogenetic trees, we can conclude that 1) directions in trophic evolution in Parnassiinae (Kreuzberg and Dyakonov 1990) are similar to those in morphological evolution; and that 2) all morphological modifications in male genitalia are synapomorphic (i.e., every group having characteristic features is monophyletic) (these morphological groups correspond to published molecular groupings – for more details about male genitalia in Parnassiinae see
Based on these conclusions, we follow the Parnassiinae classification developed by
Genus Driopa Korshunov, 1988. Diagnostic characters: primary host-plants are Fumariaceae; saccus well developed; uncus and tegumen connection solid; gnathos present; valva without harpa, no caudal branches, no chaetae; sphragis massive, sclerotized, and occupying more than 3 abdominal tergits. In cladograms as a separate clade or sister to Koramius (see for example
Genus Parnassius. Diagnostic characters: primary host-plants are Crassulaceae, secondary host-plants are Fumariaceae; uncus and tegumen connection not solid; saccus reduced; valva with a massive harpa; sphragis not massive, occupying fewer than 3 abdominal tergits. In cladograms always as separate clade.
Genus Tadumia Moore, 1902. Diagnostic characters: host-plants are Crassulaceae or Fumariaceae; gnathos present; valva divided into sclerotised basal lobe with long harpa and non-sclerotised caudal lobe; uncus massive, sclerotised; sphragis weakly sclerotised. In cladograms always close to Koramius.
Genus Kreizbergius Korshunov, 1990. Diagnostic characters: host-plants are Scrophulariaceae; sphragis absent; uncus with only one lobe; uncus and tegumen connection solid; valva small, scale-shaped; phallus very long and curved (almost 90 degrees). In cladograms always as separate clade.
Genus Sachaia Korshunov, 1988. Diagnostic characters: primary host-plants are Fumariaceae; subscaphium present; uncus consisting of two separate lobes; saccus massive and with two parts; sphragis skinny. In cladograms most often clustering with Kreizbergius, but sometimes as separate clade (
Genus Koramius Moore, 1902. Diagnostic characters: host-plants are Fumariaceae; gnathos absent; uncus with direct connection to tegumen; harpa located in the median part of valva; valva with sets of chaetae; saccus cylindrical; sphragis tube-shaped. In cladograms always as separate clade. Divided into two subgenera: Koramius – with a two-lobed tube-shaped sphragis, short uncus (shorter than tegumen) and short harpa (less than half of valval length); Kailasius Moore, 1902 – with a one-lobe tube-shaped sphragis, long uncus (equal length or longer than tegumen) and long harpa (longer than half of valval length). See male genitalia in Figs
Male genitalia of Driopa mnemosyne (Linnaeus, 1758) from different locations: A – Kirghizia, Suusamyrtoo Mts., right shore of Suusamyr River, 2300 m; B – Russia, Caucasus, Teberda; C – EU, Slovakia, Boleraz; D – Kirghizia, Kirghiz Mts., Issyk-Ata Valley, 2000 m; E – Kirghizia, Alai Mts., Kul-Kush Valley; F – Kazakhstan, Transili Alatau Mts., Bolshaya Almaatinka Valley, 2500 m.
Male genitalia of Parnassius apollonius (Eversmann, 1847) from the same locality. Kirghizia, Suusamyr Valley, west shore of Karakol River, 2200–2500 m.
Male genitalia of Parnassiinae. A – Koramius (Kailasius) autocrator, Tajikistan, Sarezskoe Lake; B – Lingamius szechenyii (Frivaldsky, 1886), China, Qinhai near Tsa-Ka; C, D – Sachaia tenedius (Eversmann, 1851), Russia, Transbaikal, Mondy (in C subscaphius removed); E – Kreizbergius simonius (Staudinger, 1889), lectotype; F – Koramius (Koramius) staudingeri (Bang-Haas, 1882), lectotype; G – K. stoliczkanus (C. & R. Felder, 1865), Kashmir.
Koramius charltonius (Gray, 1853) (=Parnassius charltonius) is one of the most enigmatic butterfly species from the Central Asian mountains and as such it appears to be of never-ending interest to butterfly collectors. Almost every known population has been described as a standalone subspecies, making one wonder whether this is a case of taxonomic inflation, similar to what is seen in other species of the genus Parnassius (
The mountains of Central Asia represent a biodiversity hotspot between rather poor arid lowlands and quite rich mountainous habitats (
Although there are several different species concepts, there is some agreement among practicing systematists on what constitutes a species. However, the same cannot be said for subspecies, but as the discussion about subspecies concept is not the aim of our study, we follow the one by
We use Koramius charltonius as an example of a species with many described subspecies of uncertain or unstable status. From Central Asia, subspecies have been described from almost all locations where the species was found in the last 20 years. The distribution is probably wider than known and could correspond to a distribution of its host-plants, Corydalis stricta Steph. & Fisch. and C. gortschakovii Schrenk. (Fumariaceae) (
Koramius charltonius’ (Fig.
Koramius charltonius (Gray, 1853). Imago (mistericus, paratype, KP689312), genitalia (Chitral, Baroghil Pass), host-plant (Corydalis gortschakovii), habitat (Tajikistan, West Pamirs, 35–45 km N of Khorog, the type locality of safronovi) and sequenced specimens.
Many described taxa belonging to K. charltonius are very difficult to place into a logical system because differences described in original descriptions begin to disappear in larger series. The wing pattern and even wing venation in Parnassiinae are very variable and cannot be used for taxon differentiation without statistical support (
Distribution map of Koramius charltonius (Gray, 1853) in Central Asia. Black circles – type localities of described subspecies with no DNA sampled by us (mined from GenBank); black circles with grey ovals – type localities with sampled specimens (paratypes or topotypes). For numbers see Table
Material used in the morphometric analysis.
|N||Subspecies||No. males||No. females||Locality|
|1||aenigma||29||8||East Aiai, Kok-Su/Kysyl-Su (Kyrgyzstan)|
|2||alrashid||13||13||East Alai, Gulcha (Kyrgyzstan)|
|6||flaugeri||13||13||Jammu and Kashmir (India)|
|7||kabiri||15||14||Vantch Mts., Gushkhon (Tajikistan)|
|8||ljudmilae||15||12||Hissarsky Mts. (Tajikistan)|
|10||mistericus||-||11||Dunkeldyk Lake env. (Tajikistan)|
|12||platon||16||15||Turkestansky Mts., Isfana (Kyrgyzstan)|
|13||romanovi||29||27||Aram-Kungei, Transalai Mts (Kyrgyzstan)|
|14||safronovi||2||7||Khorog env. (Tajikistan)|
|15||sochivkoi||26||26||North-East Alai, Ak-Bura (Kyrgyzstan)|
|16||vaporosus||42||38||Darvaz, Viskharvi Pass; Darvaz, Obimazor River (Tajikistan)|
|17||varvara||10||10||Tian-Shan, Dzhaman-Too Mts. (Kyrgyzstan)|
|18||voigti||11||11||Panjshir Valley (Afghanistan)|
Sampled specimens of K. charltonius (see Fig.
|N||“Subspecies” ID||Sequence ID||GenBank Accession Number||Locality||Haplotype (#, group)|
|1||anjuta||CHAR22/14||KP689301||Tajikistan, East Pamir, Mynkhadzhir mountains, 4200 m (PARATYPE)||14, C|
|2||aenigma||CHAR004/14||KP689302||Kyrgyzstan, eastern part of Alai valley, confluence of Rivers Kok-Suu and Kyzyl-Suu, 3000 m.||15, B|
|3||alrashid||IDPARN12/14||KP689303||Kyrghyzstan, Alai Mts., Gulcha River valley, 3200 m.||17, B|
|4||eugenia||PARN222/14||KP689308||Tajikistan, NW Pamirs, Muksu River valley, 3600 m.||10, A|
|5||ljudmilae||PARNB09/14||KP689304||Tajikistan, Gissarsky Mts., Shakhrinav environs, 3700 m.||12, A|
|6||mistericus||KORB0018/12||KP689312||Tajikistan, Sarykolsky Mts., Dunkeldyk lake, 4200 m. (PARATYPE)||14, C|
|7||marusya||PARN221/14||KP689307||Tajikistan, Darvazsky Mts., Obikhingou River valley, 3700 m (PARATYPE)||9, A|
|8||platon||KORB0017/12||KP689311||Kyrgyzstan, Turkestansky Mts., Sarkat River valley, 1500 m. (PARATYPE)||26, D|
|9||romanovi||IDPARN04/14||KP689305||Kyrgyzstan, Alai Mts., Kichik-Alai gorge, Isfairamsay River valley, 3500 m.||16, B|
|10||safronovi||CHAR003/14||KP689309||Tajikistan, W. Pamirs, 35- 40 km N of Khorog, 3800 m (HOLOTYPE)||21, C|
|11||sochivkoi||CHAR012/14||KP689306||Kyrgyzstan, Alai Mts., Kichik-Alai gorge, Ak-Bura River valley, 3500 m. (PARATYPE)||17, B|
|12||varvara||KORB0016/12||KP689310||Kyrgyzstan, Dzhaman-Too Mts., Karasu River valley, 2900 m (PARATYPE)||1, A|
|13||voigti||KORB0021/12||KP689313||Afghanistan, Koh-i-Baba Mts.||29, D|
aenigma Dubatolov & Milko, 2003. Type locality: “Kyrghyzstan, east from the Alai valley, right bank of the Kyzylsuu river at its confluence with the Koksuu river, loess-pebble bluff, 2900 m”.
alrashid Churkin & Pletnev, 2012. Type locality: “NE Alai, Gulcha river”.
anjuta Stshetkin & Kaabak, 1985. Type locality: “East Pamir, Mynkhadzhim mountain gorge, 4200 m”.
deckerti Verity, 1907. Type locality: “Kaschmir, 4200–4800 m”.
eugenia Churkin, 2009. Type locality: “Tadjikistan, Muksu R.”.
ljudmilae Lesin & Kaabak, 1991. Type locality: “Tadzhikskaya SSR, Gissarsky Mts., Diakhan-Dara upper stream 40 km N of Shakhrinav, 3700 m”.
marusya Churkin & Pletnev, 2012. Type locality: “Tajikistan, Obikhingou r.”.
mistericus Kaabak, Sotchivko & Titov, 1996. Type locality: “Tadjikistan, South-East Pamirs, western spurs of Sarykolsky Mts Ridge, Dunkeldyk Lake, rocky crumbling slope, 4300 m above sea level”.
platon Sotchivko & Kaabak, 2011. Type locality: “SW Kyrgyzstan, Turkestansky Mts. Range, Sarkat River, 1500 m above s. l.”.
romanovi Grum-Grshimailo, 1885. Type locality: “Transalai” (by the lectotype). Actually type locality shown in the original description located in mountainous pass near Katta-Karamuk village, South Kyrgyzstan.
safronovi Korb, Shaposhnikov, Zatakovoy & Nikolaev, 2013. Type locality: “Tajikistan, South-West Pamir, Shakhdarinsky Mts., 35–45 km N of Khorog, 3700 m”.
sochivkoi Churkin, 2009. Type locality: “North-East Alai, Ak-Bura River, 2600–2700 m”.
vaporosus Avinov, 1913. Type locality: “Bukhara, Darvaz, Viskharvi Pass”.
varvara Churkin, 2009. Type locality: “Kyrgyzstan, Dzhaman-Too Mts., Karasu R., …2900 m”.
Recently the phylogeography of several Parnassiinae species has been studied using DNA markers (
The species K. charltonius is currently of high commercial interest; the new subspecies are becoming valuable commodities on the market. We stress three main aims of this study: 1) to reveal the relationships of individual populations of K. charltonius throughout the Central Asian mountains (western part of the distribution); 2) to investigate the phylogeographic pattern in the area; and 3) to use the available information to try to test the validity of the described subspecies.
For this study, we selected populations known from Central Asia (we use this term to refer to the countries of Kazakhstan, Kyrgyzstan, Tajikistan, Turkmenistan, and Uzbekistan). No special attention to selection of phenotypes has been done; rather, we paid much more attention to the reliability of localities for our specimens than to the external features, based on the principle that if a specimen originates from the locality which is now treated as an area of a certain subspecies, it must have this subspecies’ features (including the COI sequence). All sequenced specimens are depicted in Fig.
Samples of K. charltonius from GenBank used in current study.
|N||“Subspecies” ID||GenBank Accession Number||Locality||Haplotype (#, group)|
|1||aenigma||KJ961627||Kyrgyzstan: Irkeshtam, Chinese Kyzyl-Su River||29, D|
|2||alrashid||KJ961626||Kyrgyzstan: Gulcha River (2200–2500 m), East Alai||28, D|
|3||amabilis||KJ961611||India: Bhaga valley, Baralacha La (4400 m)||7, A|
|4||anjuta||KJ961616||Tajikistan: Mynkhadzhyr Mts., East Pamir||1, A|
|5||charltonius||KJ961609||China: Mandhata Mt. (4800-5200 m), W. Tibet||30, -|
|6||charltonius||DQ407774||Pakistan: Satrapa Pass||22, D|
|7||deckerti||KJ961610||India: Lamayuru (3900 m), Ladakh||5, A|
|8||deckerti||KJ961613||India: S. of Khardung La (5000 m), Ladakh||3, A|
|9||ducalis||KJ961618||Pakistan: Birmoglasht (2500-4000 m), Chitral||1, A|
|10||eisnerianus||KJ961612||India: N. of Tsokar (4600 m), Ladakh||6, A|
|11||ella||KJ961617||Pakistan: Soost (3000-4000 m), Khudabaad Mts.||2, A|
|12||ella||KJ961615||Pakistan: Phakora to Naltar (2000-3200 m), Gilgit||1, A|
|13||eugenia||KJ961629||Tajikistan: Muksu River, Peter I Mts.||23, D|
|14||flaugeri||KJ961614||Pakistan: Babusar Pass||4, A|
|15||kabiri||KJ961625||Tajikistan: Gyshkhun (3000 m), Vanch Mts., W. Pamir||13, C|
|16||kabiri||KJ961624||Tajikistan: Kuh-i-Lal (3500 m), W. Pamir||14, C|
|17||ljudmilae||KJ961630||Tajikistan: Karatag River basin (3500 m), Ghissar||24, D|
|18||marusya||KJ961623||Tajikistan: Obikhingou River (2200–2400 m), Darvas||23, D|
|19||platon||KJ961621||Tajikistan: Yangiaryk (1800-1900 m)||25, D|
|20||romanovi||KJ961620||Kyrgyzstan: Aram Kungei River (3500 m), Trans-Alai||22, D|
|21||sakai||AM231451||Sonamarg, Kashmir, India||8, A|
|22||sochivkoi||KJ961631||Kyrgyzstan: Ak-Bura River (2600–2700 m), NE Alai||27, D|
|23||varvara||KJ961628||Kyrgyzstan: Karasu River (3000 m), Dzhaman-Too Mts., Tian-Shan||29, D|
|24||vaporosus||KJ961622||Tajikistan: Obimazor (3500 m), Mazorsky Mts., Darvas||19, C|
|25||vaporosus||EF473789||No data||14, C|
|26||voigti||KJ961619||Afghanistan: Panjshir valley (3500-4000 m), Parvan Prov.||11, A|
Samples of other Koramius and other outgroup species from GenBank used in this study.
|N||Species ID||GenBank Accession Number||Country|
|5||K. charltonius (wrongly determined as inopinatus)||EF473790||Afghanistan|
The type specimens of the following K. charltonius taxa have been studied: aenigma (paratypes, 1 ♂, 1 ♀), anjuta (holotype ♂, paratypes 3 ♂, 3 ♀), charltonius (syntype 1 ♀), eugenia (paratypes 2 ♂, 2 ♀), ljudmilae (holotype ♂), mistericus (holotype ♂, paratypes 3 ♂, 3 ♀), platon (paratypes 2 ♂, 2 ♀), romanovi (paralectotypes 1 ♂, 1 ♀), safronovi (complete type series), sochivkoi (paratypes 1 ♂, 1 ♀), vaporosus (lectotype ♂), varvara (paratype ♂).
Additional material studied (all topotypes): aenigma (12 ♂, 6 ♀), alrashid (2 ♂), anjuta (14 ♂, 6 ♀), eugenia (2 ♂), ljudmilae (2 ♂, 1 ♀), mistericus (32 ♂, 12 ♀), platon (26 ♂, 26 ♀), romanovi (112 ♂, 43 ♀), sochivkoi (12 ♂, 2 ♀), vaporosus (56 ♂, 42 ♀); also non-topotypic material have been studied for morphometric analysis (see Table
The genitalia in Parnassiinae do not show much variability and the majority of the taxa were described using only wing pattern. Therefore we selected ten wing pattern characteristics for males and females for 18 subspecies described from Central Asia. In total, we measured 347 males and 315 females (Table
DNA extraction, PCR and sequencing were carried out according to the protocols described in
For the sequence processing, phylogenetic analysis, and producing a timed-tree we used the software BioEdit (
We ended up excluding several samples from
Ordination plots of CCA analyses of variables of wing pattern elements on subspecies of K. charltonius. A) males, B) females. Plotted are ordination axes 1 and 2. Continuous variables are plotted as arrows, dummy variables as white triangles. Black triangles represent the centroid of subspecies.
The Bayesian inference (BI) and maximum likelihood (ML) trees of Koramius charltonius and closely related taxa. Outgroups are shown in grey. The colours represent the four haplotype lineages.
The majority of our measured wing characteristics poorly separated subspecies of K. charltonius. The only excluded variable from the final model was the colour of MRedSp in males (for single-term tests, see Table
Single term tests of wing pattern analyses for males and females of K. charltonius from CCA with 999 Monte Carlo permutations.
According to our molecular analyses (Fig.
K. ch. charltonius from Tibet, i.e. the nominotypical subspecies, is sister to all the other populations of K. charltonius, which are split into two clades (ML tree; this is also supported by the network analysis and thus we prefer this arrangement), or with K. ch. voigti sister to other Central Asian populations (BI tree). With this exception, the BI and ML trees are very similar despite low support for the main clades. One clade is formed by populations from east Tajikistan and Kashmir (haplotype group A in Fig.
Haplotype network analysis shows high haplotype divergence (Fig.
The messiness of the phylogenetic relationships among these putative taxa is reflected in the messiness of the systematics of this species. Clearly, using the phylogenetic tree, it is impossible to circumscribe separate subspecies and this result supports our conclusions about the unrealistic number of subspecies of K. charltonius. On the other hand, the haplotype network visualizes the grouping and connections between the populations and it can be used for taxonomic interpretations, as it was done recently, for example, by
Even when we take into account the differences between ML and BI trees, we see that the number of described subspecies is not realistic. Moreover, our results do not support the traditional division of described subspecies into groups (sensu
The genetic distances were marginally positively correlated with geographic distances (Mantel test, r = 0.271, p = 0.071), but they were not correlated with morphological distances (r = 0.051, p = 0.324). Even when we made comparison with only males (r = -0.021, p = 0.567) or females (r = 0.082, p = 0.253), we did not find any correlation. Also, when comparing morphology distance matrices for males and females, we did not find a correlation (r = -0,001, p = 0.481). Thus the differences between these subspecies are not real; for some specimens it is not possible to identify to which subspecies of K. charltonius they belong without having geographical data. Recognizable phenotypes are present only in subspecies romanovi, varvara, and platon. Additionally, it is possible to identify some groups of subspecies according to the wing pattern.
Based on the results of our examinations, we propose a preliminary subspecific division of K. charltonius in Central Asia. It is possible that it will be necessary to further reduce the number of subspecies in the future because there is no clear pattern in phylogenetic trees. We used the haplotype network and morphometric data as a basis for the following division:
varvara, characterised by island-distribution in the Inner Tian-Shan (it is possible that this subspecies name will be synonymized after further investigation).
romanovi, characterized by distribution in the Alai and Transalai Mts. The southern border of this subspecies is on the high-mountainous plain starting from the southern slope of the Transalai Mts., the eastern border is at the beginning of the Zeravshan River Valley. Synonyms: aenigma Dubatolov & Milko, 2003, syn. n., sochivkoi Churkin, 2009, alrashid Churkin & Pletnev, 2012, syn. n.
platon, characterized by distribution in the north-eastern limit of the species range in Central Asia (Turkestansky Mts.) and by the mid-altitude mountainous vertical distribution (1500–2100 m). This kind of vertical distribution is unusual for this species and makes a notable ecological isolation from other known populations.
ljudmilae, characterized by the distribution in the south-eastern limit of the species distribution in Central Asia (Hissarsky Mts.). There are long distances (over 200 km) to its other closest known populations.
vaporosus, characterized by distribution in east, central and north-west Pamir and Darvaz. The borders of this subspecies are Alichur River in the south, highland plains of East Pamir in the east, the high-mountainous plain starting from the southern slope of Transalai Mts. in the north, and the eastern slopes of Darvaz Mts. in the west. Synonyms: marusya Churkin & Pletnev, 2012, syn. n., eugenia Churkin, 2009, syn. n., anjuta Stshetkin & Kaabak, 1985, syn. n., mistericus Kaabak, Sotchivko & Titov, 1996, syn. n.
voigti, distributed in West Pamir and Afghan Badakhshan. The northern border is in the Akademii Nauk Mts. and adjacent glaciers. Synonym: safronovi Korb, Shaposhnikov, Zatakovoy & Nikolaev, 2013, syn. n.
We present the phylogenetic relationships of populations of Koramius charltonius based on one mitochondrial gene from the western part of its range. Whereas two haplotype groups (A and D) are widely distributed throughout the range, two other haplotype groups (B and C) indicate a separation of larger historical areas by vicariance, first in isolated sections of the westernmost parts of the area (isolation of platon and ljudmilae), then separation of voigti, and then separation of the southern (vaporosus) and northern (romanovi) branches with deckerti in between.
The observed pattern suggests that the species had a wide distribution in Central Asia in the past, at least up to the southern ridges of Tian-Shan (probably wider). The first Pleistocene stage divided the area of the K. charltonius ancestor into several large areas separated by high mountain ridges, which were covered by glaciers: e.g., Alai and Transalai, Sary-Kol etc. (
These results indicate a shrinking of the original range, with western populations becoming isolated due to the gradual drying of the mountains rather than dispersal. Also, it is evident that the eastern populations are not really genetically isolated and thus the majority of the described subspecies lack genetic justification.
We found very weak support for the traditional division (
Many of the new subspecies descriptions are frequently inadequately erected, based rather on partial data than on systematic comparison of large series and using adequate analyses. Even worse, many of such subspecies might have been established only to increase the price of the insect specimens on the market. However, these taxa are still valid in the sense of the zoological nomenclature. Taxonomists must acknowledge every described taxon, they must place it in its correct position or synonymize it, and we have done our best with this difficult case based on all the currently available data.
We are thankful to Mr V.V. Titov (Zheleznodorozhnyi, Russia) for granting most of used material for DNA studying; we also thankful to Mr P. Egorov (Almaty, Kazakhstan) for the same. We thank Dr A.V. Sviridov (Moscow State University, Zoological Museum, Moscow, Russia), Dr S.Yu. Sinev and Dr A.L. Lvovsky (Zoological Institute of the Russian Academy of Sciences, St.-Petersburg, Russia), Dr W. Mey (Museum für Naturkunde, Berlin, Germany) for giving access to study of curated collections. For providing materials for study we are greatly indebted also to Mr D.A. Pozhogin (Nizhny Novgorod, Russia), Mr A.A. Shaposhnikov (Podolsk, Russia), Mr A.A.Zatakovoi (Nizhny Novgorod, Russia), Dr L.V. Kaabak (Moscow, Russia). We are indebted to M. Nuss and Z. Varga for reviews of an earlier version, which helped us improve our text. We are tankful to J. Rota for final editing of the manuscript. M. Sweney helped with linguistic corrections. The sequencing was supported by Czech Science Foundation (GA CR) (14-36098G), by the University of South Bohemia (168/2013/P) and by the Nizhny Novgorod State University (contract 22-11-2013/c).