Short Communication |
Corresponding author: Vladimir A. Lukhtanov ( lukhtanov@mail.ru ) Academic editor: Vladimir Gokhman
© 2017 Vladimir A. Lukhtanov, Nazar A. Shapoval.
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Citation:
Lukhtanov VA, Shapoval NA (2017) Chromosomal identification of cryptic species sharing their DNA barcodes: Polyommatus (Agrodiaetus) antidolus and P. (A.) morgani in Iran (Lepidoptera, Lycaenidae). Comparative Cytogenetics 11(4): 759-768. https://doi.org/10.3897/compcytogen.v11i4.20876
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DNA barcoding has been suggested as a universal tool for molecular species identification; however, it cannot be applied in cases when morphologically similar species share their DNA barcodes due to the common ancestry or mitochondrial introgression. Here we analyze the karyotype of Polyommatus (Agrodiaetus) morgani (Le Cerf, 1909) from the region of its type locality in the southern Zagros Mountains in Iran, provide first chromosomal evidence for P. (A.) antidolus (Rebel, 1901) in Iran and demonstrate that these two species can be easily identified through analysis of their karyotypes whereas they share their mitochondrial barcodes.
Ancestral polymorphism, biodiversity, chromosomes, chromosomal fusion/fission, cryptic species, cytogenetics, DNA barcoding, incomplete lineage sorting, karyosystematics, mitochondrial introgression, nomenclature, species identification, taxonomy
Cryptic species, morphologically indistinguishable or highly similar biological entities, represent a substantial portion of plant and animal diversity, and therefore the search for these species is important for taxonomic, ecological and evolutionary studies (
Polyommatus (Agrodiaetus) antidolus (Rebel, 1901), P. (A.) kurdistanicus (Forster, 1961) and P. (A.) morgani (Le Cerf, 1909), a complex of three closely related allopatric species distributed in east Turkey as well as in west and central Iran (Fig.
Map of Iran showing the type locality of Polyommatus (Agrodiaetus) morgani and the localities of the analyzed specimens of P. (A.) morgani and P. (A.) antidolus. 1 type locality of P. (A.) morgani, “Deh-Tcheshma” (Deh Cheshme near Farsan, Chaharmahal and Bakhtiari Province) 2P. (A.) morgani, n=27, vic. Sibak, Esfahan Province 3P. (A.) morgani, n=27-29, 25 km E of Mahabad, W. Azerbaijan Province 4P. (A.) morgani, n=27-29, 15 km W of Mahabad, W. Azerbaijan Province 5P. (A.) antidolus, n=39-41, Seir, 4 km S of Urmia, W. Azerbaijan Province.
The butterflies were collected in 2016 in north-west and central Iran: in a mountain valley between Fereydunshahr and Sibak (locality 2), in the vicinity of Darman (25 km E of Mahabad) (locality 3), in the vicinity of Khalifen (15 km W of Mahabad) (locality 4) and in Seir (near Urmia) (locality 5) (Fig.
List of studied material (17 specimens). Asterisks indicate unsequenced specimens. Collectors: E. Pazhenkova (EP), N. Shapoval (NS), V. Lukhtanov (VL).
Field Code | GenBank number | Taxon | Chromosome number (n) | Locality | Altitude | Date | Collectors/ References |
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Q055* | morgani ♂ | n=27 | Iran, Esfahan Prov., Sibak (N32°55'; E50°04') | 2700 m | 02.08.2017 | EP, NS, VL | |
Q060* | morgani ♂ | n=27 | Iran, Esfahan Prov., Sibak (N32°55'; E50°04') | 2700 m | 02.08.2017 | EP, NS, VL | |
Q150 | MG457163 | morgani ♂ | n=28-29 | Iran, W. Azerbaijan Prov., vic. Darman, 25 km E of Mahabad (N36°45'; E45°52') | 1900–2000 m | 10.08.2017 | EP, NS, VL |
Q170 | MG457164 | morgani ♂ | n=27 | Iran, W. Azerbaijan Prov., vic. Darman, 25 km E of Mahabad (N36°45'; E45°52') | 1900–2000 m | 10.08.2017 | EP, NS, VL |
Q171 | MG457165 | morgani ♂ | n=27 | Iran, W. Azerbaijan Prov., vic. Darman, 25 km E of Mahabad (N36°45'; E45°52') | 1900–2000 m | 10.08.2017 | EP, NS, VL |
Q181 | MG457166 | morgani ♂ | n=28 | Iran, W. Azerbaijan Prov., vic. Darman, 25 km E of Mahabad (N36°45'; E45°52') | 1900–2000 m | 10.08.2017 | EP, NS, VL |
Q196 | MG457167 | morgani ♂ | n=27-28 | Iran, W. Azerbaijan Prov., vic. Khalifen, 15 km W of Mahabad (N36°45'; E45°32') | 2100–2200 m | 11.08.2017 | EP, NS, VL |
Q197 | MG457168 | morgani ♂ | n=27 | Iran, W. Azerbaijan Prov., vic. Khalifen, 15 km W of Mahabad (N36°45'; E45°32') | 2100–2200 m | 11.08.2017 | EP, NS, VL |
Q198 | MG457169 | morgani ♂ | n=28-29 | Iran, W. Azerbaijan Prov., vic. Khalifen, 15 km W of Mahabad (N36°45'; E45°32') | 2100–2200 m | 11.08.2017 | EP, NS, VL |
Q237 | MG457170 | antidolus ♂ | n=40-41 | Iran, W. Azerbaijan Prov., vic. Seir, Urmia (N37°28'; E45°02') | 1750 m | 14.08.2017 | EP, NS, VL |
Q238 | MG457171 | antidolus ♂ | n=39-40 | Iran, W. Azerbaijan Prov., vic. Seir, Urmia (N37°28'; E45°02') | 1750 m | 14.08.2017 | EP, NS, VL |
Q239 | MG457172 | antidolus ♂ | n=39 | Iran, W. Azerbaijan Prov., vic. Seir, Urmia (N37°28'; E45°02') | 1750 m | 14.08.2017 | EP, NS, VL |
AY557093 | antidolus ♂ | n=42 | Turkey, Hakkari Prov., Dez Çay | 1500 m | 22.07.1999 |
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AY557095 | antidolus ♂ | n=ca44 | Turkey, Hakkari Prov., Haruna Geçidi, SE Yüksekova | 2000 m | 21.07.1999 |
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AY557108 | kurdistanicus ♂ | n=ca>55 | Turkey, Van Prov., Erek Dagi | 2200 m | 25.07.1999 |
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AY557074 | kurdistanicus ♂ | n=ca54-56 | Turkey, Van Prov., Çatak | 1600–1900 m | 25.07.1999 |
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AY496762 | kurdistanicus ♂ | n=62 | Turkey, Van Prov., Çatak | July 2001 |
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Divergence between COI sequences. The numbers of base differences per site between sequences are shown. The shared barcodes (uncorrected COI p-distance = 0) are shown in bold.
1 | 2 | 3 | 4 | 5 | 6 | 7 | 8 | 9 | 10 | 11 | 12 | 13 | 14 | 15 | |
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(1) AY557095antidolus | |||||||||||||||
(2) AY557089antidolus | 0.0015 | ||||||||||||||
(3) AY557108kurdistanicus | 0.0015 | 0 | |||||||||||||
(4) AY557074kurdistanicus | 0.0015 | 0 | 0 | ||||||||||||
(5) AY496762kurdistanicus | 0.0015 | 0 | 0 | 0 | |||||||||||
(6) Q150 morgani | 0.0015 | 0 | 0 | 0 | 0 | ||||||||||
(7) Q170 morgani | 0.0029 | 0 | 0 | 0 | 0 | 0 | |||||||||
(8) Q171 morgani | 0.0029 | 0.0015 | 0.0015 | 0.0015 | 0.0015 | 0.0015 | 0.0015 | ||||||||
(9) Q181 morgani | 0.0015 | 0 | 0 | 0 | 0 | 0 | 0 | 0.0015 | |||||||
(10) Q196 morgani | 0.0015 | 0 | 0 | 0 | 0 | 0 | 0 | 0.0015 | 0 | ||||||
(11) Q197 morgani | 0.0015 | 0 | 0 | 0 | 0 | 0 | 0 | 0.0015 | 0 | 0 | |||||
(12) Q198 morgani | 0.0015 | 0 | 0 | 0 | 0 | 0 | 0 | 0.0015 | 0 | 0 | 0 | ||||
(13) Q237 antidolus | 0.0015 | 0 | 0 | 0 | 0 | 0 | 0 | 0.0015 | 0 | 0 | 0 | 0 | |||
(14) Q238 antidolus | 0.0015 | 0 | 0 | 0 | 0 | 0 | 0 | 0.0015 | 0 | 0 | 0 | 0 | 0 | ||
(15) Q239 antidolus | 0.0015 | 0 | 0 | 0 | 0 | 0 | 0 | 0.0015 | 0 | 0 | 0 | 0 | 0 | 0 |
In order to investigate the topotypical population of P. (A.) morgani, we first searched for it in its exact type locality in “Deh Tcheshma” (mountain area near the village Deh Cheshme, close to the city Farsan, Chaharmahal and Bakhtiari Province, Iran) (locality 1 in Fig.
Male wing pattern of P. (A.) morgani, P. (A.) antidolus and P. (A.) kurdistanicus. aP. (A.) morgani, Iran, Kordestan Province, Senandaj, 1800 m, 20 July 2000, leg. P. Hofmann bP. (A.) antidolus Turkey, Hakkari Province, Ogul-Tal, 1500–1900 m, 1 August 1984, leg. Schurian cP. (A.) kurdistanicus Turkey, Van Province, 10 km S of Van, 1900–2100 m, 10 August 1978, leg. Görgner.
Karyotype of P. (A.) morgania Q060, MI, n=27 b Q055, MI, n=27 c Q170, prometaphase I, n=27 d Q171, MI, n=27 e Q197, prometaphase I, n=27 f Q196, MI, n=27 g Q196, MII, n=27 h Q196, MII, n=28 i Q196, MII, n=28. Bar = 10 μ.
Finally, in three specimens collected in Seir (near Urmia, locality 5 in Fig.
Karyotypes of P. (A.) morgani and P. (A.) antidolus. aP. (A.) morgani, Q150, MII, n=28 bP. (A.) morgani, Q150, MII, n=29 cP. (A.) antidolus, Q239, MI, n=39, squash preparation dP. (A.) antidolus, Q239, MI, n=39, intact metaphase plate eP. (A.) antidolus, Q237, MII, n=40 fP. (A.) antidolus, Q237, MII, n=41. Scale bar = 10 μ.
We are grateful to Wolfgang ten Hagen for providing information about sampling localities of P. (A.) morgani in Iran. The complete financial support for this study was provided by the grant from the Russian Science Foundation N 14-14-00541 to The Zoological Institute of the Russian Academy of Sciences. The work was partially performed using equipment of the Centre for Molecular and Cell Technologies and Department of Entomology of St. Petersburg State University. A part of this equipment was purchased with support of the St. Petersburg University grant 1.40.490.2017.