Research Article |
Corresponding author: Vladimir A. Lukhtanov ( lukhtanov@mail.ru ) Academic editor: Christina Nokkala
© 2017 Vladimir A. Lukhtanov, Alexander V. Dantchenko.
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:
Lukhtanov VA, Dantchenko AV (2017) A new butterfly species from south Russia revealed through chromosomal and molecular analysis of the Polyommatus (Agrodiaetus) damonides complex (Lepidoptera, Lycaenidae). Comparative Cytogenetics 11(4): 769-795. https://doi.org/10.3897/CompCytogen.v11i4.20072
|
Finding a new species is a rare event in easy-to-see and well-studied organisms like butterflies, especially if they inhabit well-explored areas such as the Western Palaearctic. However, even in this region, gaps in taxonomic knowledge still exist and here we report such a discovery. Using a combined analysis of chromosomal and molecular markers we demonstrate that Polyommatus blue populations from Daghestan (South Russia), previously identified as P. aserbeidschanus, represent in fact a new species which is described here as P. australorossicus sp. n. We also show that the enigmatic Polyommatus damonides described as a form of Polyommatus damone and later considered as an entity similar to P. poseidon or P. ninae is conspecific with a taxon previously known as P. elbursicus. As a result of our study, we propose several taxonomic changes within the P. damonides species complex and suggest the following new combinations: P. damonides elbursicus Forster, 1956, comb. n. and P. damonides gilanensis Eckweiler, 2002, comb. n.
Ancestral polymorphism, biodiversity, chromosomes, chromosomal fusion/fission, COI , cryptic species, DNA barcoding, incomplete lineage sorting, inverted meiosis, karyosystematics, molecular phylogenetics, mitochondrial introgression, phylogeography, speciation
Agrodiaetus , a subgenus of the species-rich Palaearctic genus Polyommatus Latreille, 1804 (
Species identification in Agrodiaetus is complicated. The morphology of male genitalia is uniform for most of the species. With a few exceptions, it can help to separate groups of species (
Here we use a combination of molecular mitochondrial (COI) and nuclear chromosomal (karyotype) markers to analyze the taxa and populations close to Polyommatus damonides (= lineage VIII in
Specimens examined (Supplementary Table 1, Fig.
The Bayesian tree of studied Polyommatus samples based on analysis of the cytochrome oxidase subunit I (COI) gene. Numbers at nodes indicate Bayesian posterior probability. I, II and III are recovered haplogroups of the P. damonides species complex. Polyommatus shamil, phenotypically similar to P. australorossicus, but genetically distant, was used to root the tree.
Karyotypes were obtained from 157 adult males representing eight species and were processed as previously described (
Standard COI barcodes (658-bp 5’ segment of mitochondrial cytochrome oxidase subunit I) were studied. COI sequences were obtained from 30 specimens representing the P. damonides species group and from 9 samples of P. shamil (Dantchenko, 2000) which was selected as outgroup. Legs were sampled from these specimens, and sequence data from the DNA barcode region of COI were obtained at the Canadian Centre for DNA Barcoding (CCDB, Biodiversity Institute of Ontario, University of Guelph) using protocols described in
We also used 28 published COI sequences (
The barcode analysis involved 67 COI sequences. Sequences were aligned using the BioEdit software (
157 specimens were karyotyped (Supplementary Table 1, Figs
P. ninae (Fig.
At the MI/MII stages, the number of chromosome elements was found to vary from n=ca32 to n=34-36 in 21 studied specimens from different localities, with n=33 and n=34 as distinct modal numbers. All chromosome elements formed a gradient size row. The species seemed to be polymorphic for at least one chromosomal fusion/fission resulting in specimens possessing 33 bivalents (homozygotes for fused chromosomes) (Fig.
Karyotypes of P. ninae, P. aserbeidschanus and P. australorossicus sp. n. Trivalents are indicated by arrows. a P. ninae, sample 2014VL34, MI, n=34 b P. ninae, sample 2014VL39, MI, n=33 c P. ninae, sample 2014VL33, MI, n=32 bivalents + 1 trivalent (heterozygote for fusion/fission) d P. ninae, sample 2014VL60, n=32 bivalents + 1 trivalent (heterozygote for fusion/fission) e P. ninae, sample 2014VL33, MII, n=33 f P. aserbeidschanus, sample 05A406, MI, n=32 g P. aserbeidschanus, sample 05A387, MI, n=33 h P. aserbeidschanus, sample 05A387, MI, n=33 i P. australorossicus sp. n., sample DK-27-97-1, MI, n=23. Bar = 10 μ.
P. aserbeidschanus (Fig.
At the MI/MII stages, the number of chromosome elements was found to vary from n=32 to n=37 in 17 studied specimens from different localities, with n=33 as a modal number. MI/MII metaphases consisted of elements of progressively decreasing size.
P. australorossicus sp. n. (Fig.
At the MI/MII stages, the haploid chromosome number n = 23 was found in 6 studied individuals. Elements were found to form a gradient size row in which the largest element was approximately 5 times larger than the smallest element. In two specimens, the diploid chromosome number was estimated as 2n = 46 in male atypical meiosis. In the sample DK-7-97 we counted approximately n = ca22 and in the sample from Chonkatau we counted approximately n = ca24 at the MI stage. The last two counts were done with an approximation due to the overlapping of some bivalents, therefore interpretation of these deviating numbers (a real variation or a mistake of counting) is difficult.
P. damonides damonides from Azerbaijan and Armenia (Fig.
At the MI/MII stages, the haploid chromosome number n = 18 was found in 10 studied individuals. Elements formed a gradient size row in which the largest element was approximately 2-2.5 times larger than the smallest element. In two specimens, the diploid chromosome number was determined as 2n = 36 in male atypical meiosis.
Karyotype of P. damonides from Ordubad vicinity in Nakhchivan (Azerbaijan) and Meghri vicinity in Armenia. a sample 005A14K, three MI plates (two from polar view and one from equatorial view) displaying n=18 b sample 005A14K, MI, n=18 c sample 005A14K, MII, n=18 d sample 2014VL04, male atypical meiosis, 2n=36. Bar = 10 μ.
P. damonides from Iran (previously known as P. elbursicus) (Fig.
At the MI/MII stages, the haploid chromosome number n = 18 was found in 26 studied individuals. Elements constituted a gradient size row in which the largest element was approximately 2-2.5 times larger than the smallest element. In 7 specimens, the diploid chromosome number was determined as 2n = 36 in male atypical meiosis. Thus, the karyotype of these samples from Iran is indistinguishable from the karyotype of the samples of P. damonides from Azerbaijan and Armenia.
Karyotype of P. damonides from Iran (previously known as P. elbursicus). a sample E234, MI, n=18 b sample E193, MI, n=18 c sample E460, MI, n=18 d sample E237, MI, n=18 e sample E459, MI, n=18 f sample E193, MII, n=18 g sample J573, two sister MII plates, n=18 h sample E234, male atypical meiosis, 2n=36. Bar = 10 μ.
P. damonides elbursicus (Forster, 1956) (Fig.
At the MI/MII stages, the haploid chromosome number n = 17 was found in four studied individuals. Elements formed a gradient size row in which the largest element was approximately 2-2.5 times larger than the smallest element. In the sample VL311, the diploid chromosome number was determined as 2n = 34 in male atypical meiosis.
Karyotype of P. damonides elbursicus from Elburs Mts (north Iran). a sample M804, MI, n=17 b sample VL302, two MI plates, each displaying n=17 c sample VL302, diakinesis – early prometaphase, n=17 d sample VL302, prometaphase, n=17 e sample VL302, MII, n=17. Bar = 10 μ.
P. damonides gilanensis Eckweiler, 2002 (Fig.
At the MI/MII stages, the number of chromosome elements was found to vary from n=18 to n=19 in three studied specimens collected in the type-locality of this taxon. Elements formed a gradient size row in which the largest element was approximately 2-2.5 times larger than the smallest element. The population was found to be polymorphic for a chromosomal fusion/fission resulting in specimens possessing 18 bivalents (homozygotes for fused chromosomes), 17 bivalents + 1 trivalents (heterozygotes for fusion/fission) (Fig.
Karyotype of P. damonides gilanensis from its type-locality. Trivalents and triple chromatid are indicated by arrows. a sample J111, MI, n=18 b sample J111, MI, n=18 c sample J111, MII, n=18 d sample J112, MI, n=19 e sample J112, MI, n=19 f sample J112, MI, n=19. Bar = 10 μ.
P. zarathustra (Fig.
At the MI/MII stages, the number of chromosome elements was found to vary from n=20-21 to n=24 in 6 studied specimens from different localities, with n=22 as a modal number. Elements formed a gradient size row in which the largest element was approximately 5 times larger than the smallest element. The species seemed to be polymorphic for several, still unrecognized chromosomal rearrangements resulting in chromosome number variation.
Karyotypes of P. zarathustra, P. arasbarani, P. lukhtanovi and P. shamil. a P. zarathustra, sample Z401, MI, n=24 b P. arasbarani arasbarani, sample N98, MI, n=24 c P. arasbarani neglectus, sample Q457, MI, n=ca25-26 d P. lukhtanovi, sample F875, MI, n=22 e P. lukhtanovi, sample H717, MI, n=21 f P. shamil, sample F958, MI, n=17. Bar = 10 μ.
P. arasbarani arasbarani (Fig.
At the MI/MII stages, the number of chromosome elements was found to vary from n=23-24 to n=25 in 6 studied specimens, most likely due to polymorphism for a single chromosomal fusion/fission. Elements formed a gradient size row in which the largest element was approximately 5-6 times larger than the smallest element.
P. arasbarani neglectus Dantchenko, 2000 (Fig.
At the MI stage, the number of chromosome elements was determined to be n=25 in the sample B447. In the samples KA-95-99, 2001-Q456 and 2001-Q457 the number of elements was estimated with an approximation as n=24-25 and n=25-26 due to the overlapping of some bivalents. In the sample KA-98-99, the diploid chromosome number was estimated as 2n = ca 48. Elements formed a gradient size row in which the largest element was approximately 5-6 times larger than the smallest element.
P. lukhtanovi (Fig.
At the MI/MII stages, in 28 of 33 studied specimens the haploid chromosome number was determined as n=22. In one of these 28 specimens atypical meiosis displayed 2n=44. In 3 of 33 studied samples the haploid chromosome number was determined as n=21, and in two samples intraindividual variation in the number of elements was observed: n=21-22. We interpret this result as an evidence for polymorphism for a single fusion/fission resulting in in specimens possessing n=21 (Fig.
P. shamil (Fig.
At the MI/MII stages, in all 12 studied specimens the haploid chromosome number was determined as n=17. In three of these 12 specimens atypical meiosis displayed 2n=34. Elements formed a gradient size row in which the largest element was approximately 2 times larger than the smallest element.
The COI barcode analysis revealed three major, highly supported clusters within the studied samples (Fig.
Distribution of COI haplogroups (I – III) and haploid chromosome numbers (n) in the P. damonides species complex.
Within its distribution range P. ninae demonstrates a diversity of COI haplotypes; however, no distinct intraspecific sublineages were discovered. Polyommatus ninae firuze (Carbonell, 1993) described from Turkey (Gümüşhane, Kelkit) shows no differentiation from topotypical populations from Armenia, and in our opinion should be considered no more than a synonym of P. ninae.
The specimens of P. aserbeidschanus collected in the type locality of this taxon “Armenia, mts. Zangezur, pag. Kadzharantz, pr. Mts. Kapudzhich” (
On the tree obtained, the samples of P. australorossicus were intermixed together with the samples of P. ninae. Although no shared haplotypes were found, the uncorrected p-distances between the P. ninae and P. australorossicus samples were quite low varying from 0.2 % to 1.4 %. Thus, both P. ninae and P. australorossicus appeared on the tree as genetically undifferentiated, non-monophyletic assemblages.
The second lineage (haplogroup II) is represented by samples of P. damonides, P. elbursicus, P. elbursicus gilanensis, P. zarathustra, P. arasbarani and P. lukhtanovi. This is the southern lineage of the P. damonides complex distributed in the south-eastern part of Nakhchivan (Ordubad district, Azerbaijan), south-eastern part of Armenia (Meghri district), south-eastern part of Azerbaijan (Talysh) and Iran (Fig.
The third lineage (haplogroup III) (south-western group) is represented by samples of P. pierceae. This lineage inhabits the south–eastern part of Turkey (Fig.
The P. damonides species complex demonstrates a high rate of karyotype evolution resulting in a great interspecific diversity of chromosome numbers (from n=17 to n 34-36) (Figs
There are two possible ways of the first meiotic division in a cell with the fusion/fission trivalent: (i) resulting in a daughter cell containing two smaller chromosomes and a daughter cell containing one larger chromosome, and (ii) resulting in two daughter cells, each containing an element consisting of three triple chromatids (
The fact that the discovered fusions/fissions can exist in populations in both homo- and heterozygous conditions indicates, most likely, that these rearrangements can pass through meiosis and are not strongly underdominant. Previously, the low or no underdominance of chromosomal fusions/fissions was demonstrated for butterflies of the genus Leptidea Billberg, 1820 (
Thus, the fixed differences in karyotype are not only (syn)apomorphic characters demonstrating that chromosomal races represents distinct phylogenetic lineages, i.e. species from the point of view of phylogenetic species concepts, but also indirect evidence for at least partial reproductive isolation.
The studied complex demonstrates a high level of chromosomal differentiation between taxa and a relatively low level of differentiation with respect to the mitochondrial COI gene, with many distinct taxa intermixed on the COI tree obtained (Fig.
In case of the pair P. ninae – P. aserbeidschanus which are indistinguishable in both molecular and chromosomal characters, we can also hypothesize that these two nominal taxa are conspecific. These two taxa have been long time considered as distinct species because of a wrong assumption about their karyotypic differentiation (
However, P. ninae and P. aserbeidschanus are not identical with respect to their morphology and ecological preferences. Male specimens of P. aserbeidschanus (mostly collected around the type locality in South Zangezur Range) have specific dark brown coloration on the wing underside, blue ground color with violet tint on the wing upperside and significantly smaller size compared with the males of P. ninae. Polyommatus aserbeidschanus is known only from the subalpine belt of the South Zangezur mountain area and connected trophically with the Astragalus species preliminary determined as Astragalus prilipkoanus (sectio Incani) (Fabaceae) (
A similar case is found in the pair P. zarathustra – P. arasbarani. These two taxa are allopatric, and similar with respect to morphology, karyotypes and COI barcodes. However, they are differentiated with respect to ecological preferences: P. zarathustra is associated with dry areas in central Iran, whereas P. arasbarani is associated with meadow-like biotopes in subalpine zone of the north-west Iran. Polyommatus arasbarani neglectus is known only from low and middle altitude on southern slopes of the Meghri mountain range, it inhibits dry glades and clearance in an oak forest belt and trophically connected with astragalus species preliminary determined as Astragalus fedorovi (sectio Incani) (Fabaceae). This ecological differentiation does not allow synonymaizing these taxa, and further studies are required to clarify this situation, too.
Lycaena Damone var. Damonides Staudinger, 1899 is the oldest taxon described within the studied complex. Therefore analysis of its identity is of great importance for solving nomenclatural problems within the group. The taxon was described as a form of Polyommatus damone (hypothesis 1) and later considered as an entity close to P. poseidon (
According to our observations P. damonides damonides inhabits tragacanth and Paliurus plant communities from 1000m. alt. (in Armenia) to 2100 m. alt. (in Nakhchivan, vicinity of Ordubad) and is trophically connected with Astragalus ordubadensis (sectio Incani) (Fabaceae) which is endemic of South Zangezur mountain range. It is also important to note that in Meghri-Ordubad region we have found sympatry/syntopy for the species pairs P. arasbarani neglectus/P. damonidesdamonides, P. arasbarani neglectus/P. aserbeidschanus and P. damonides/P. aserbeidschanus.
(Fig.
(BOLD process ID BPAL2013-13; GenBank accession number MG243366).
ACATTATATTTTATTTTTGGAATTTGAGCAGGAATAGTAGGAACATCCNTAAGAATTTTAATTCGTATAGAATTGAGAA CTCCTGGATCCTTAATTGGAGATGATCAAATTTATAACACTATTGTTACAGCTCATGCATTTATTATAATTTTTTTTATA GTTATACCTATTATAATCGGAGGATTTGGTAACTGATTAGTTCCTTTAATATTAGGGGCACCTGATATAGCCTTTCCACG ACTAAATAATATAAGATTCTGATTATTACCGCCATCATTAATACTACTAATTTCCAGAAGAATTGTAGAAAATGGAGCAG GAACAGGATGAACAGTTTACCCCCCACTTTCATCTAATATTGCACATAGAGGATCATCTGTAGATTTAGCAATTTTCTCT CTTCATTTAGCAGGAATTTCTTCAATTTTAGGAGCAATTAATTTTATTACAACTATTATTAATATACGGGTAAATAATTT ATCTTTTGATCAAATATCATTATTTATTTGAGCAGTGGGAATTACAGCATTATTATTACTTTTATCTTTACCTGTATTAG CTGGAGCAATTACCATATTATTAACTGATCGAAATCTTAACACCTCATTCTTTGATCCAGCTGGTGGAGGAGATCCAATT TTATATCAACATTTA
9 males. (1) BOLD process ID BPAL2011-13, field # CCDB-17947_B04; karyotype preparation DK-34-1-97; Russia, Caucasus, Daghestan, Gimrinsky Range, Gunib, 1800 m, 15 August 1997, A. Dantchenko leg. (2) BOLD process ID BPAL2012-13, field # CCDB-17947_B05; karyotype preparation DK-34-2-97, n=ca23; Russia, Caucasus, Daghestan, Gimrinsky Range, Gunib, 1800 m, 15 August 1997, A. Dantchenko leg. (3) BOLD process ID BPAL2014-13, field # CCDB-17947_B07; karyotype preparation DK-7-97, n=ca22; Russia, Caucasus, Daghestan, Gimrinsky Range, Gunib, 1800 m, 12 August 1997, A. Dantchenko leg. (4) karyotype preparation DK-23-97, n=23, 2n=46; Russia, Caucasus, Daghestan, Gimrinsky Range, Gunib, 1800 m, 15 August 1997, A. Dantchenko leg. (5) karyotype preparation DK-30-97, n=23; Russia, Caucasus, Daghestan, Gimrinsky Range, Gunib, 1800 m, 15 August 1997, A. Dantchenko leg. (6) karyotype preparation DK-23-97-3, n=23; Russia, Caucasus, Daghestan, Gimrinsky Range, Gunib, 1800 m, 14 August 1997, A. Dantchenko leg. (7) karyotype preparation DK-23-97-4, 2n=ca46; Russia, Caucasus, Daghestan, Gimrinsky Range, Gunib, 1800 m, 14 August 1997, A. Dantchenko leg. (8) karyotype preparation DK-27-97-2, n=23; Russia, Caucasus, Daghestan, Gimrinsky Range, Gunib, 1800 m, 14 August 1997, A. Dantchenko leg. (9) karyotype preparation n=?24; Russia, Caucasus, Daghestan, Chonkatau, V. Tikhonov leg. All paratypes are deposited in the Zoological Institute of the Russian Academy of Science (St. Petersburg).
(no DNA, no karyotype). 10 males: Russia, Caucasus, Daghestan, Gimrinsky Range, Gunib, 1450–1950 m, 11–16 August 1997, A. Dantchenko leg.
Males. Forewing length 16.5–18.5 mm.
Upperside: Ground colour bright glossy violet blue with narrow black marginal line, marginal part of forewings and hindwings slightly dusted with black scales, discal strokes absent, veins darkened distally, costal area of the forewings white, basal part of fringe dark grey on forewings, light grey on hindwings, distal part white.
Underside: Forewing ground colour grey, submarginal row blurred, but clear visible; discoidal strokes black, bordered with white; postdiscal rows of black spots bordered with white, 80% males have basal black spots; hindwing ground colour grey with ocherous tint, basal area with strong greenish suffusion; discal stroke less prominent than on forewings; postdiscal row of black spots bordered with white, submarginal and antemarginal marking not strong but clear visible; submarginal row bordered distally with reddish brackets, more pronounced to anal end of row; white streak sharp, equal in width; basal half of fringes pale grayish on fore- and hindwings, distal part white.
Females remains unknown.
Genitalia. The male genitalia have a structure typical for other species of the subgenus Agrodiaetus (
Stony steppe and dry meadows from 1500 up to 2000 m a.s.l. Flight period: mid-July to end of August, in a single generation. The new species flights syntopically and synchronously with P. shamil but on average about one decade earlier. Host plant is preliminary determined as Astragalus buschiorum (Fabaceae). Hibernation as first instar larvae.
Phenotypically P. (A.) australorossicus sp. n. is practically indistinguishable from allopatric closely related P. ninae, P. aserbeidschanus and P. lukhtanovi but the ground colour of the underside of the hindwings is grey in the new species, with ocherous tint, not light or dark brown. The new species differs from sympatric (syntopic and synchronous) P. shamil (Fig.
Specimens of Polyommatus (Agrodiaetus) australorossicus sp. n. and P. (A.) shamil. Both samples collected in Gunib (Russia, Caucasus, Daghestan, Gimrinsky Range, 1600-1800 m), 14 August 1997, by A. Dantchenko. a, b upperside (a) and underside (b) of the holotype of Polyommatus (Agrodiaetus) australorossicus sp. n. DK-27-97, n=23; arrow indicates basal black spot c, d upperside (c) and underside (b) of the paratype of Polyommatus (Agrodiaetus) shamil, CCDB-17947_B11, DK-97-18, n=17, 2n=34. Bar = 10 mm.
Pubescence of the anterior part of the forewing upper surface. a, b, c, d the anterior part of the forewing upper surface possesses a strong white pubescence in the area bordered by the costal margin (CM) and the veins R and R2+3, i.e. this area is densely covered with relatively long white hairs (a P. aserbeidschanus b P. ninae, c P. arasbarani neglectus d P. australorossicus) e, f the white pubescence of the anterior part of the forewing upper surface is strongly reduced and limited to the only costal margin, the white hairs are short (e P. ciscaucasicus f P. shamil) g the white pubescence of the anterior part of the forewing upper surface is reduced, not dense (P. damonides) h schematic picture showing the venation of the forewing in Polyommatus and the photographed area (shaded). Bar = 3 mm.
Genetically P. australorossicus and P. shamil are not close. They belong to two different species groups within the subgenus Agrodiaetus: to P. carmon group (P. australorossicus) and to P. cyaneus group (P. shamil).
The new species differs drastically from the genetically most closely related P. ninae and P. aserbeidschanus by its karyotype (by at least 9 fixed chromosomal fusions/fissions).
The new species is similar (but not identical) to P. lukhtanovi (n=21-22) and P. pierceae (n=22) with respect to the chromosome number. However, it differs from these species by COI barcodes and represents a different lineage of evolution within the P. damonide complex.
The name australorossicus is an adjective of the masculine gender. This species name originates from the Latin words “australis” (south) and “rossicus” (Russian).
We propose the following taxonomic arrangement of the P. damonides species complex (chromosome numbers are in parentheses):
i. Polyommatus (Agrodiaetus) ninae lineage
P. (A.) ninae (Forster, 1956) (Agrodiaetus transcaspica ninae Forster, 1956; =Agrodiaetus ninae firuze Carbonell, 1993) (n=33-35)
P. (A.) aserbeidschanus (Forster, 1956) (Agrodiaetus transcaspica aserbeidschana Forster, 1956) (n=32-37)
P. (A.) australorossicus, sp. n. (n=23)
ii. Polyommatus (Agrodiaetus) damonides lineage
P. (A.). damonides (Staudinger,1899)
P. (A.) damonides damonides (Staudinger, 1899) (Lycaena Damone var. Damonides Staudinger, 1899) (n=18)
P. (A.) damonides elbursicus (Forster, 1956), comb. n. (Agrodiaetus transcaspica elbursica Forster, 1956) (n=17)
P. (A.) damonides gilanensis Eckweiler, 2002, comb. n. (Polyommatus (Agrodiaetus) elbursicus gilanensis Eckweiler, 2002) (n=18-19)
P. (A.) lukhtanovi (Dantchenko, 2005) (Agrodiaetus lukhtanovi Dantchenko, 2005) (n=21-22)
P. (A.) zarathustra Eckweiler, 1997 (Polyommatus (Agrodiaetus) zarathustra Eckweiler, 1997) (n=20-24)
P. (A.) arasbarani (Carbonel & Naderi, 2000)
P. (A.) arasbarani arasbarani Carbonel & Naderi, 2000 (Agrodiaetus arasbarani Carbonel & Naderi, 2000) (n=24-25)
P. (A.) arasbarani neglectus Dantchenko, 2000 (Polyommatus (Agrodiaetus) zarathustra neglectus Dantchenko, 2000; = Polyommatus (Agrodiaetus) arasbarani ihmal Koçak & Kemal, 2008) (n=24-26)
iii. Polyommatus (Agrodiaetus) pierceae lineage
P. (A.) pierceae (Lukhtanov & Dantchenko, 2002) (Agrodiaetus pierceae Lukhtanov & Dantchenko, 2002) (n=22)
Comment. The name Polyommatus (Agrodiaetus) arasbarani ihmal was suggested by
The volume 48 of Neue Entomologische Nachrichten with description of Polyommatus (Agrodiaetus) zarathustra neglectus Dantchenko, 2000 was published and distributed in May 2000. Additional studies are required to clarify what taxon (described by Dantchenko or described by Stradomsky and Arzanov) was published first. Until this situation is resolved in a future revision, we see no other way than to use P. (A.) arasbarani neglectus Dantchenko, 2000 as a valid name.
We thank Mark Kalashian (Scientific Center of Zoology and Hydrobiology, Yerevan, Armenia), Vahe Sargsyan (Arevik National Park, Meghri, Armenia), David Tarkhnishvili (Ilia State University, Tbilisi, Georgia), and Valentin Tikhonov (Russia, Pyatigorsk) for generous hospitality and help in butterflies collecting. Valentin Tikhonov provided also several samples important for this study. We also thank George Fayvush (Institute of Botany, Yerevan, Armenia) for supplying us botanic literature and Andrei Sytin (Komarov Botanical Institute, St. Petersburg, Russia) for help in determination of Astragalus species. The financial support for this study was provided by the grant N 14-14-00541 from the Russian Science Foundation to the Zoological Institute of the Russian Academy of Sciences. The work was partially performed using equipment of the ‘Chromas’ Core Facility, the Centre for Molecular and Cell Technologies and the Department of Entomology of St. Petersburg State University. Some of this equipment was partly purchased with support of the St. Petersburg University grant 1.40.490.2017. 350.
Table S1
Data type: PDF table.
Explanation note: Chromosome numbers of the studied Polyommatus (Agrodiaetus) samples with their IDs, localities, dates and collectors.