Analyses of morphology and mitochondrial DNA reveal a deep split within Parnassius charltonius

The article presents the synthesis of mtDNA, morphological and geographical data on different groups of taxa united under the name Parnassius charltonius Gray, [1853]. Parnassius romanovi gruM-grShiMailo, 1885 stat. nov. constitutes a bona species distributed in Alai, Ghissar, West Pamirs/Badakhshan and the SW edges of Tian-Shan. It is characterized by a reduced uncus (which is shorter than the tegumen) and a reduced third submarginal spot on the HW underside. The status of the taxa deckerti Verity, 1907 and voigti Bang-haaS, 1927 is questionable and needs further clarification. Especially the latter taxon could constitute a separate species. Comparative analysis of mtDNA and morphological data makes it possible to interpret the action of evolutionary factors in the history of the taxa. Introduction: The butterfly family Papilionidae has recently been the object of numerous molecular and phylogenetic studies, many of which included representatives of Parnassius latreille, 1804 (oMoto et al., 2004; Katoh et al., 2005; Nazari et al., 2007; Gratton et al., 2008; MiChel et al., 2008; oMoto et al., 2008; TodiSCo et al., 2010, 2012; CondaMine et al., 2012). During the same period, field studies resulted in the discovery of a number of new taxa in the subgenus Kailasius Moore, 1902, which includes some of the most spectacular members of Parnassius latr. One new species (P. davydovi Churkin, 2006) and at least six new subspecies of P. charltonius Gray, [1853] were found and described recently, the distribution area of the latter being significantly enlarged as a result. During these studies, it was found that the structure of the † genitalia of P. charltonius Gray differs significantly from that of other representatives of Kailasius Moore (Churkin, 2006) and, moreover, that different charltonius taxa have remarkably different genitalia (Churkin & PletneV, 2012). The necessity to confirm and extend these findings prompted us to undertake an in-depth comparative study of taxa united under the name „Parnassius charltonius Gray“ and to this end, we chose to associate morphological analyses with the sequencing of DNA. Such combined analyses have been published for various Vertebrate complexes, but are not yet commonplace for insects, including butterflies (for Parnassiinae, the study of the Zerynthia polyxena complex by Zinetti et al., 2013, constitutes a recent exception). The primary goal of this work was to clarify the status of the northern populations (united under the oldest available name romanovi gruM-grShiMailo, 1885), but our results, which reveal a deep, ancient split within P. charltonius Gray, also raise questions about the microevolution, adaptive radiation and multidimensional analysis of the species involved. At the same time, it should be noted that ascertaining the status of individual subspecies is beyond the limits of the present article, if only since this will require systematic work based on lectotype designations that are necessary to restrict the type localities. Unfortunately, that remains to be done for many Parnassius taxa, including P. charltonius Gray (only in the romanovi group the taxonomic work was finished recently by the senior author of the present paper). Abbreviations: FW fore wing, HW hind wing, TL type locality, mtDNA mitochondrial DNA I. Morphological studies 1. Groups of taxa and distribution The distribution area of P. charltonius Gray (fig. F1) extends from Western Tibet (close to its frontiers) and Western Nepal to Bamyan in Afghanistan; its northern limits are the Alai and Turkestan ranges and the south-western part of Inner Tian-Shan. It is not known from the main territory of Tian-Shan, West Ghissar or the western edges of Afghanistan. According to different authors, the subspecific structure of P. charltonius Gray is formed of three or four groups (plate 1-2). KreuzBerg (1985) confirmed the existence of four groups (“deckerti”, “charltonius”, “romanovi” and “voigti”), but distinctive characters were not listed. roSe & WeiSS (2011) used three groups only, uniting the romanovi and voigti groups, but again without a detailed analysis of distinctive features. The romanovi-group occupies the northern part of the species range, i.e Ghissar, Alai, Transalai (including the upper stream of the Chinese Kyzyl-Su river.), Darwas, West Pamirs and the SE part of Tian-Shan. In total, 11 subspecies are known, mainly with deep hiatuses in colouration; the detailed situation will be discussed separately. For the sake of comparative analysis of group distinctions, it is important to note the clinal structure of the characters found among populations of vaporosus aVinoV, 1913 kabiri eiSner & nauMann, 1980 (Churkin & PletneV, 2014). While butterflies from the Vanch range (West Pamirs) are the most variable ones within this complex, that population has no possibility to contact individuals from the voigti and deckerti groups. In contrast, butterflies from the Dzhilandy or Ishkashimsky ranges (which lie closest to the distribution areas of the voigti and deckerti groups) exhibit less variaAtalanta 45 (1-4): 97-125, Würzburg (Sept. 2014), ISSN 0171-0079