vavilovВавиловский журнал генетики и селекцииVavilov Journal of Genetics and Breeding2500-3259Institute of Cytology and Genetics of Siberian Branch of the RAS10.18699/vjgb-26-74vavilov-5226Research ArticleГЕНЕТИКА РАСТЕНИЙPLANT GENETICSМолекулярно-цитогенетический анализ некоторых видов Picea из Евразии и Восточной АзииMolecular cytogenetic analysis of some Picea species from Eurasia and East AsiaГорячкинаО. В.GoryachkinaO. V.

Красноярск

Krasnoyarsk

kvitko@ksc.krasn.ruБадаеваЕ. Д.BadaevaE. D.

Москва

Moscow

ЩербаньА. Б.ShcherbanA. V.

Новосибирск

Novosibirsk

МуратоваЕ. Н.MuratovaE. N.

Красноярск

Krasnoyarsk

Институт леса им. В.Н. Сукачева Сибирского отделения Российской академии наук – обособленное подразделение Федерального исследовательского центра «Красноярский научный центр Сибирского отделения Российской академии наук»РоссияV.N. Sukachev Institute of Forest of the Siberian Branch of the Russian Academy of Sciences, Federal Research Center “Krasnoyarsk Science Center of the Siberian Branch of the Russian Academy of Sciences”Russian FederationИнститут общей генетики им. Н.И. Вавилова Российской академии наук; Институт молекулярной биологии им. В.А. Энгельгардта Российской академии наукРоссияN.I. Vavilov Institute of General Genetics of the Russian Academy of Sciences; Engelhardt Institute of Molecular Biology of the Russian Academy of SciencesRussian FederationФедеральный исследовательский центр Институт цитологии и генетики Сибирского отделения Российской академии наукРоссияInstitute of Cytology and Genetics of the Siberian Branch of the Russian Academy of SciencesRussian Federation202626082026305735746Copyright © Горячкина О.В., Бадаева Е.Д., Щербань А.Б., Муратова Е.Н., 20262026Горячкина О.В., Бадаева Е.Д., Щербань А.Б., Муратова Е.Н.Goryachkina O.V., Badaeva E.D., Shcherban A.V., Muratova E.N.Данная работа распространяется под лицензией Creative Commons Attribution 4.0.This work is licensed under a Creative Commons Attribution 4.0 License.https://vavilov.elpub.ru/jour/article/view/5226

Представлены результаты молекулярно-цитогенетического исследования семи видов Picea, распространенных на территории Евразии и Восточной Азии, методом флуоресцентной гибридизации in situ

(FISH) и флуоресцентного окрашивания DAPI. Кластеры рибосомных генов 45S и 5S рРНК на хромосомах P. meyeri и P. purpurea картированы впервые, P. abies, P. obovata, P. schrenkiana, P. crassifolia, P. omorika изучались из новых местопроизрастаний. Все виды показали сходное распределение локусов 5S рДНК на обоих плечах крупной метацентрической хромосомы III: более интенсивный сигнал наблюдался на длинном плече, частично перекрываясь с сигналом 45S рДНК, минорный сайт 5S рДНК был локализован терминально на коротком плече. Сигналы 45S рДНК разной интенсивности были выявлены на 7–9 парах хромосом в кариотипах исследуемых  видов. Яркие (мажорные) сигналы локализованы интеркалярно и совпадали с постоянными вторичными перетяжками. Их количество на гаплоидный геном составило: у P. omorika – 8, у P. abies и P. obovata – 6, у P. schrenkiana, P. crassifolia, P. purpurea и P. meyeri – 5. Выявлено также несколько минорных сайтов 45S рДНК на хромосомах Picea, не описанных ранее в литературе. Добавочные (В) хромосомы, обнаруженные у некоторых проростков P. obovata, P. meyeri и P. purpurea, не несли локусов рибосомных генов 45S и 5S рРНК. На хромосомах ели наблюдались интенсивно флуоресцирующие DAPI-позитивные полосы в интеркалярных районах, а у некоторых хромосом окрашивалась также область центромеры. На хромосомах I, III и X пар удалось выявить несколько постоянных позитивных DAPI-блоков, имеющих сходную локализацию у всех изученных видов. FISH с зондами 45S и 5S рДНК в сочетании с окрашиванием DAPI позволяет идентифицировать практически все гомологичные пары хромосом и провести сравнение кариотипов разных видов ели. Обсуждается внутри- и межвидовое кариотипическое разнообразие в роде Picea на основе распределения локусов 5S и 45S рДНК и рисунка DAPI. 

Karyotypes of seven Picea species distributed across Eurasia and East Asia were analyzed using fluorescence in situ hybridization (FISH) and DAPI fluorescent staining. For P. meyeri and P. purpurea, the 5S and 45S rDNA sites were mapped here for the first time, while karyotypes of P. abies, P. obovata, P. schrenkiana, P. crassifolia, and P. omorika were examined from new locations. All the species studied had a similar distribution of 5S rDNA loci on both arms of the large metacentric chromosome III: a more intense signal was observed in the middle of the long arm, partially overlapping with the 45S rDNA signal, and a minor 5S rDNA site was localized terminally on the short arm. The distribution of 5S rDNA loci varied in the karyotypes of the studied species: from 7 to 9 pairs of chromosomes carried 45S rDNA signals of varying intensity. Strong (major) signals were localized intercalarily and coincided with persistent secondary constrictions. Their number per haploid genome was as follows: 8 in P. omorika, 6 in P. abies and P. obovata, and 5 in P. schrenkiana, P. crassifolia, P. purpurea, and P. meyeri. We also detected several minor 45S rDNA sites on the Picea chromosomes that had not been previously described in the literature. Supernumerary (B) chromosomes found in some seedlings of P. obovata, P. meyeri, and P. purpurea did not carry 45S and 5S rDNA loci. Intensely fluorescent DAPI-positive bands were observed in the intercalary regions of the spruce chromosomes, and the centromere regions of certain chromosomes were also stained. Several constant DAPI-positive blocks were identified on chromosomes I, III, and X, with similar locations across all studied species. Fluorescence in situ hybridization with 45S and 5S rDNA probes in combination with DAPI staining allowed us to identify virtually all homologous chromosomes and compare karyotypes of different species. The intra- and interspecific karyotypic diversity in the genus Picea is discussed considering the distribution of 5S and 45S rDNA loci and DAPI signals.

хромосомакариотипFISH45S рДНК5S рДНКокрашивание DAPIPiceachromosomekaryotypeFISH45S rDNA5S rDNADAPI stainingPiceaThis work was carried out as part of the Program of the V.N. Sukachev Institute of Forest of the Siberian Branch of the Russian Academy of Sciences, Project No. FWES-2024-0028 “Biodiversity of Siberian Forests: ecological-dynamic, genetical-breeding, physical-chemical and resource-technological aspects”. Acknowledgements. The authors thank Prof. E.A. Salina, E.M. Timonova and T.N. Kapko from Institute of Cytology and Genetics SB RAS for collaboration; N.V. Oreshkova from V.N. Sukachev Institute of Forest SB RAS for Larix sibirica DNA extraction; A.B. Bezdelev from Primorsky Aquarium FEB RAS, I.V. Tikhonova from V.N. Sukachev Institute of Forest SB RAS, Y.V. Tkac heva from Donetsk Botanical Garden and O.A. Mochalova from the Institute of Biological Problems of the North FEB RAS for supplying the seed samples. Isolation and cloning of the 5S rDNA probe were conducted in the Laboratory of molecular genetics and cytogenetics of plants, Institute of Cytology and Genetics SB RAS.This work was carried out as part of the Program of the V.N. Sukachev Institute of Forest of the Siberian Branch of the Russian Academy of Sciences, Project No. FWES-2024-0028 “Biodiversity of Siberian Forests: ecological-dynamic, genetical-breeding, physical-chemical and resource-technological aspects”. Acknowledgements. The authors thank Prof. E.A. Salina, E.M. Timonova and T.N. Kapko from Institute of Cytology and Genetics SB RAS for collaboration; N.V. Oreshkova from V.N. Sukachev Institute of Forest SB RAS for Larix sibirica DNA extraction; A.B. Bezdelev from Primorsky Aquarium FEB RAS, I.V. Tikhonova from V.N. Sukachev Institute of Forest SB RAS, Y.V. Tkac heva from Donetsk Botanical Garden and O.A. Mochalova from the Institute of Biological Problems of the North FEB RAS for supplying the seed samples. 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The authors declare that there are no conflicts of interest present.