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Молекулярно-Генетическое Исследование Некоторых Кавказских Представителей Рода Kretania Beuret, 1959 (Lepidoptera: Lycaenidae) С Описанием Нового Вида

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Молекулярно-Генетическое Исследование Некоторых Кавказских Представителей Рода Kretania Beuret, 1959 (Lepidoptera: Lycaenidae) С Описанием Нового Вида Кавказский энтомол. бюллетень 11(1): 183–187 © CAUCASIAN ENTOMOLOGICAL BULL. 2015 Молекулярно-генетическое исследование некоторых кавказских представителей рода Kretania Beuret, 1959 (Lepidoptera: Lycaenidae) с описанием нового вида Molecular genetics study of some Caucasian representatives of the genus Kretania Beuret, 1959 (Lepidoptera: Lycaenidae) with the description of a new species Б.В. Страдомский1, В.В. Тихонов2 B.V. Stradomsky1, V.V. Tikhonov2 1Институт аридных зон ЮНЦ РАН, пр. Чехова, 41, Ростов-на-Дону 344006 Россия 2Северо-Кавказский федеральный университет, ул. Пушкина, 1, Ставрополь 355009 Россия 1Institute of Arid Zones of Southern Scientific Centre, Russian Academy of Sciences, Chekhov str., 41, Rostov-on-Don 344006 Russia. E-mail: [email protected] 2North-Caucasus Federal University, Pushkin str., 1, Stavropol 355009 Russia. E-mail: [email protected] Ключевые слова: Lepidoptera, Lycaenidae, Kretania, молекулярно-генетические маркеры, новый вид. Key words: Lepidoptera, Lycaenidae, Kretania, molecular genetic markers, new species. Резюме. Проведены молекулярно-генетические Romanian specimens and belong to Kretania sephirus исследования группы Kretania pylaon (Fischer von (Frivaldsky, 1835). Waldheim, 1832) различных популяций Кавказа, Северо- Восточной Турции и Ирана. Показано, что экземпляры К настоящему времени с территории Кавказа из Турции, Южной Грузии, Армении и Талыша имеют описаны 5 таксонов группы Kretania pylaon (Fischer von близкие нуклеотидные последовательности COI-гена Waldheim, 1832). С западной части Большого Кавказа и относятся к одному виду. Старейшим пригодным были описаны Plebejus (Plebejides) pylaon abchasicus названием для него является Plebeius sephirus modicus Nekrutenko, 1975 (типовое местонахождение: Verity, 1935. Устанавливается новый статус этого Абхазия, озеро Рица), Plebejus (Plebejides) pylaon таксона – Kretania modica (Verity, 1935), stat. n. albertii Nekrutenko, 1975 (типовое местонахождение: Популяции восточной половины Большого Кавказа Теберда, Джамагат), Plebeius (Plebejides) sephirus (Дагестан, Северо-Восточный Азербайджан, Восточная kubanensis Shchurov, 1999 (типовое местонахождение: Грузия) имеют отличные от других представителей рода Анапа, Сукко). Из Южного Закавказья – Lycaena Kretania последовательности молекулярных маркеров: pylaon ordubadi Forster, 1938 и Plebejus (Plebejides) митохондриального COI и ядерного ITS2, – и выделены sephirus semiturcmenicus Bálint, 1991 (типовое в новый вид Kretania stekolnikovi sp. n. местонахождение: Армения, Веди) [Forster, 1938; Популяции с Западного Кавказа генетически не Nekrutenko, 1975; Bálint, 1991; Shchurov, 1999]. Из-за отличаются от экземпляров из Румынии и относятся к высокой степени изменчивости и отсутствия четких Kretania sephirus (Frivaldsky, 1835). морфологических различий их таксономическое Abstract. Molecular genetic investigations of the положение до сих пор остается запутанным и Kretania pylaon species group populations from the неоднозначным. Так, подвид abchasicus был отнесен Caucasus, North Eastern Turkey and Iran are made. к K. sephirus (Frivaldszky, 1835), а подвиды ordubadi и Specimens from Turkey, Southern Georgia, Armenia and alberti переподчинены иранскому виду K. zephyrinus Azerbaijan (Talysh Mountains) have similar COI nucleotide (Christoph, 1884) [Балинт и др., 1992]. Относят к sequences and belong to one species. The oldest name K. zephyrinus также популяции бабочек из Дагестана for this species is Plebeius sephirus modicus Verity, 1935. [Львовский, Моргун, 2007]. Молекулярно-генетические Species rank for this taxon is established: Kretania modica исследования и ранее позволили нам усомниться в (Verity, 1935), stat. n. Populations from eastern part of верности идентификации дагестанских популяций the Big Caucasus (Dagestan, North Eastern Azerbaijan, как K. zephyrinus [Водолажский, Страдомский, 2010]. Eastern Georgia) have different from other representatives Однако для точного выяснения таксономических of Kretania sequences of COI and nuclear ITS2 molecular взаимоотношений кавказских бабочек рода Kretania markers. New species Kretania stekolnikovi sp. n. is Beuret, 1959 нам пришлось провести дополнительные described for these populations. Populations from the исследования молекулярных маркеров различных Western Caucasus have not genetic differences from популяций с Кавказа и сопредельных территорий. 184 Б.В. Страдомский, В.В. Тихонов Рис. 1. Местонахождения изученных экземпляров Kretania. Fig. 1. Localities of studied specimens of Kretania. Материал и методы цитохромоксидазы (COI) ДНК и ядерной некодирующей последовательности internal Исследованные экземпляры хранятся в Институте transcribed spacer 2 (ITS2), а также секвенирование аридных зон ЮНЦ РАН (Ростов-на-Дону, Россия). амплифицированных фрагментов проводили Экземплярам присвоены идентификационные аналогично процедурам, описанным ранее [Wiemers et al., 2010]. музейные номера. Местонахождения изученных Для получения ПЦР-продуктов COI использовали особей представлены на рисунке 1. Информация о прямой праймер (PolF 5’-TAG CGA AAA TGA CTT TTT материале дана в таблице 1. Материал. Kretania zephyrinus: 1♂, «Iran, Khorasan, Kopet Dagh, TCT A-3’) и обратный праймер (PolR2 5’-TTG CTC CAG Quchan, 1900–2100 m, 15.06.2006, W. ten Hagen», коллекционный CTA ATA CAG GTA A-3’), для ITS2 – прямой праймер номер ILL230 (GenBank accession number для COI: KR072206, для (PiF 5’-GGG CCG GCT GTA TAA AAT CAT A-3’) и ITS2: KR072209); 1♂, «Iran, Khorasan, Bognurd [Bujnurd], 1500–1700 m, обратный праймер (PiR 5’-AAA AAT TGA GGC AGA 23.05.2007, W. ten Hagen», коллекционный номер ILL231 (GenBank CGC GAT A-3’). accession number для COI: KR072205, для ITS2: KR072208). Kretania nichollae: 1♂, «Syria, Sweida, Hauran, 800 m, 23.04.2011, Анализ первичных нуклеотидных W. ten Hagen»,коллекционный номер ILL232 (GenBank accession последовательностей проводили с использованием number для COI: KR072204, для ITS2: KR072207). программы BioEdit Sequence Alignment Editor версии ♂ Kretania sp.: 1 , «Azerbaijan, Khizi District, Gilyazi, 600–800 m., 7.0.5.3 [Hall, 1999]. 3.05.2012, V. Tikhonov», коллекционный номер ILL233; 1♂, «Georgia, Vashlovani reserve, 9.05.2014, V. Tikhonov», коллекционный номер В работе также были использовали опубликованные ILL234; 1♂, «Armenia, Aragatsotn province, Orgov, Amberd river valley, ранее нуклеотидные последовательности экземпляров 1600–1700 m, 24.06.2014, A. Choch», коллекционный номер ILL235; 1♂, c идентификационными номерами ILL090, ILL091 и «Georgia, Samtskhe-Javakheti, Adigeni municipality, Abastumani, road ILL092 (GenBank accession number для COI: HM159433, towards Zekari pass, 1500–1600 m, A. Choch», коллекционный номер ILL236; 1♂, «Azerbaijan, Talysh, Zuvand, 1600 m, 28.05.2013, V. Tikhonov», HM159434 и HM159435; для ITS2: HM175708, коллекционный номер ILL237; 1♂, «Turkey, Malatya, Kubbe Geсidi, HM175709 и HM175710 соответственно). 1600–1800 m, 16.05.2008, W. ten Hagen», коллекционный номер ILL238. Нуклеотидные последовательности обрабатывали Обработку образцов тканей изученных с применением алгоритма Minimum Evolution с экземпляров, амплификацию участков помощью программы MEGA5 [Tamura et al., 2011] и митохондриального гена первой субъединицы графически представляли в виде ME-кладограмм. Молекулярно-генетическое исследование некоторых кавказских представителей рода Kretania Beuret, 1959 Color plate 17. Рис. 4–23. Имаго Kretania. 4–13 – Kretania stekolnikovi sp. n.: 4 – голотип; 5 – аллотип; 6–13 – паратипы; 14–23 – Kretania modica stat. n. Figs 4–23. Imago of Kretania. 4–13 – Kretania stekolnikovi sp. nov.: 4 – holotype; 5 – allotype; 6–13 – paratypes; 14–23 – Kretania modica stat. n. Молекулярно-генетическое исследование некоторых кавказских представителей рода Kretania Beuret, 1959 185 Рис. 2–3. Kretania: филогенетические деревья на основе анализа различий последовательностей ДНК с применением метода минимальной эволюции (ME). 2 – COI ген; 3 – последовательность ITS2. Figs 2–3. Kretania: phylogenetic trees based on the Minimum Evolution (ME) method of analysis of distances DNA sequences. 2 – COI gene; 3 – sequence ITS2. Результаты и обсуждение COI-гена укладывается во внутривидовые рамки: 0.4–1.1%. В некоторых литературных источниках Результаты исследований свидетельствуют о [Forster, 1938; Балинт и др., 1992; Talavera et al., 2013] том, что отличия по COI-гену между экземпляром авторы относят экземпляры из Южного Азербайджана ILL091 (Северный Кавказ, Карачаево-Черкесия, и Армении к виду K. zephyrinus. Однако изучение ущелье Джамагат) и экземплярами из Румынии [Dinca COI-гена представителей K. zephyrinus из типового et al., 2011], идентифицированными как K. sephirus, местонахождения (Иран, Копет Даг) показало их составляют всего 0.2–0.4%. Таким образом, с высокой различие с закавказскими экземплярами на видовом степенью достоверности экземпляр под номером ILL091 уровне – 2–2.6% (рис. 2). Последовательности ITS2 может быть идентифицирован как K. sephirus. В то же яДНК также свидетельствуют о том, что закавказские время экземпляры ILL235–ILL238 из Южной Грузии экземпляры и K. zephyrinus образуют разные ветви и Южного Азербайджана, Армении и Турции имеют на кладограмме (рис. 3). То есть идентификация отличие по COI-гену от K. sephirus на уровне 3.2–3.8%, закавказских экземпляров как K. zephyrinus т.е. явно представляют собой отдельный вид (рис. 2). является ошибочной. В то же время необходимо Причем внутри перечисленной группы вариабельность отметить, что изученный экземпляр из Турции имеет Таблица 1. Сводные данные об изученном материале и таксономическом
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