0536-1036 Известия высших учебных заведений. Лесной журнал Lesnoy Zhurnal (Russian Forestry Journal) ФГАОУ ВО «Северный (Арктический) федеральный университет имени М.В. Ломоносова» 10.37482/0536-1036-2026-2-73-87 https://journals.narfu.ru/index.php/fj/article/view/2568 Лесное хозяйство FORESTRY Технология клонального микроразмножения реликтовых голосеменных растений Sequoia sempervirens (D. Don) Endl. Clonal Micropropagation Technology of Relict Gymnosperms Sequoia sempervirens (D. Don) Endl. Зайцева С.М. Зайцева С.М. Zaytseva Svetlana M. smzaytseva@yandex.ru 0000-0001-9137-3774 AAE-5391-2022 Болотина Е.Л. Болотина Е.Л. Bolotina Elizaveta L. lizavetarodbol@yandex.ru 0009-0007-9006-6044 Калашникова Е.А. Калашникова Е.А. Kalashnikova Elena A. kalash0407@mail.ru 0000-0002-2655-1789 Киракосян Р.Н. Киракосян Р.Н. Kirakosyan Rima N. mia41291@mail.ru 0000-0002-5244-4311 Российский государственный аграрный университет – МСХА им. К.А. Тимирязева, ул. Тимирязевская, д. 49, Москва, Россия, 127434; Московская государственная академия ветеринарной медицины и биотехнологии им. К.И. Скрябина, ул. Академика Скрябина, д. 23, Москва, Россия, 109472 (Москва, Россия) Department of Biotechnology, Russian State Agrarian University – Moscow Timiryazev Agricultural Academy, ul. Timiryazevskaya, 49, Moscow, Russian Federation, 127434; Moscow State Academy of Veterinary Medicine and Biotechnology named after K.I. Scriabin, ul. Akademika Skryabina, 23, Moscow, Russian Federation, 109472 (Moscow, Russian Federation) Российский государственный аграрный университет – МСХА им. К.А. Тимирязева, ул. Тимирязевская, д. 49, Москва, Россия, 127434 (Москва, Россия) Department of Biotechnology, Russian State Agrarian University – Moscow Timiryazev Agricultural Academy, ul. Timiryazevskaya, 49, Moscow, Russian Federation, 127434 (Moscow, Russian Federation) Институт физиологии растений им. К.А. Тимирязева, ул. Ботаническая, д. 35, Москва, Россия, 127276 (Москва, Россия) Department of Сell Biology and Biotechnology, K.A. Timiryazev Institute of Plant Physiology, Russian Academy of Sciences, ul. Botanicheskaya, 35, Moscow, Russian Federation, 127276 (Moscow, Russian Federation) 15 04 2026 2026 2 73 87 12 02 2025 19 05 2025 18 05 2025 © Зайцева С.М., Болотина Е.Л., Калашникова Е.А., Киракосян Р.Н., 2026 2026 Зайцева С.М., Болотина Е.Л., Калашникова Е.А., Киракосян Р.Н. Zaytseva S.M., Bolotina E.L., Kalashnikova E.A., Kirakosyan R.N. CC BY 4.0 https://journals.narfu.ru/index.php/fj/article/view/2568

Sequoia sempervirens (D. Don) Endl. – самое высокое реликтовое растение, способное накапливать уникальные вторичные метаболиты, которые могут найти применение в фармакогнозии. Секвойя вечнозеленая репродуктивно уязвима, поэтому особую актуальность приобретают методы биотехнологии для создания генетических банков и биоресурсных коллекций in vitro. Объектом исследования служили черенки S. sempervirens, заготовленные с дерева, произрастающего в фондовой оранжерее Главного ботанического сада им. Н.В. Цицина РАН (Москва). В работе изучали связь минерального и гормонального составов питательной среды, а также расположения части побега, с которой изолирован черенок, и эффективности размножения секвойи in vitro. Ступенчатая стерилизация 0,1%-м раствором хлорида ртути в течение 18 мин приводила к получению хорошо растущей стерильной культуры (более 80 %). Установлено, что первоначально необходимо выращивать черенки на безгормональной питательной среде, содержащей минеральные соли по MS, а в дальнейшем на среде, в состав которой включены 2 мг/л 2,4-дихлорфеноксиуксусной кислоты + 0,5 мг/л 6-бензиламинопурина + 0,5 мг/л нафталинуксусной кислоты либо 2 мг/л 2,4-дихлорфеноксиуксусной кислоты + 2 мг/л N6-(дельта-2-изопентенил)-аденина для размножения. Для укоренения целесообразно переносить регенеранты на питательную среду с 2 мг/л 2,4-дихлорфеноксиуксусной кислоты + 0,5 мг/л 6-бензиламинопурина + 0,5 мг/л нафталинуксусной кислоты в составе. После формирования клонами корневой системы они помещаются в почвенный субстрат под полиэтиленовое укрытие при 16-часовом фотопериоде для адаптации к условиям ex vitro.

Sequoia sempervirens (D. Don) Endl. is the tallest relict plants capable of accumulating unique secondary metabolites that can be used in pharmacognosy. Given the documented reproductive vulnerability of the coast redwood, biotechnological methods for establishing in vitro genetic banks and bioresource collections are increasingly relevant for the preservation of its gene pool. This study utilized nodal segments of S. sempervirens harvested from a donor tree maintained in the stock greenhouse of the N.V. Tsitsin Main Botanical Garden of the Russian Academy of Sciences (Moscow). The research investigates the effects of mineral and hormonal nutrient media compositions, as well as the explant’s original position on the shoot, on in vitro propagation efficiency. Stepwise sterilization using a 0.1 % (w/v) mercuric chloride solution for 18 min resulted in a successful sterile culture rate exceeding 80 %. Findings indicate that explants should initially be cultured on a hormone-free Murashige and Skoog (MS) basal medium, followed by a proliferation medium supplemented with either 2 mg/L 2,4-D + 0.5 mg/L BAP + 0.5 mg/L NAA or 2 mg/L 2,4-D + 2 mg/L 2iP for effective multiplication. For rooting, it is advisable to transfer regenerants to a medium containing 2 mg/L 2,4-D + 0.5 mg/L BAP + 0.5 mg/L NAA. Following the development of the root system, the resulting clones are prepared for ex vitro transfer. Acclimatization is conducted in a soil substrate under a polyethylene cover with a 16-hour photoperiod to support the transition to ex vitro conditions.

 Ключевые слова Sequoia клоны реликтовые голосеменные растения in vitro Keywords Sequoia sempervirens clones relict plants in vitro Коллектив авторов выражает признательность агроному Главного Ботанического сада им. Н.В. Цицина РАН Кучерову Антону Валерьевичу за предоставление растительного материала. Работа выполнена при финансовой поддержке РНФ в рамках научного проекта № 24-76-00070 «Получение клеточных культур in vitro реликтовых и находящихся под угрозой исчезновения голосеменных растений рода Sequoia и изучение биологической активности ее метаболитов» (РГАУ – МСХА им. К.А. Тимирязева). The team of authors expresses its gratitude to Anton V. Kucherov, agronomist of the N.V. Tsitsin Main Botanical Garden of the Russian Academy of Sciences, for providing plant material. The study was carried out as part of the research project No. 24-76-00070. 1. Болотина Е.А., Зайцева С.М., Голиванов Я.Ю., Таций Г.Р., Бисембаев Т.Н. 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Available at: https://www.treesandshrubsonline.org/articles/sequoiadendron/sequoiadendron-giganteum/ (accessed 21.06.25). 16. Clark J.W. Natural Decay Resistance of the Heartwood of Coast Redwood Sequoia sempervirens (D. Don) Endl. Forest Products Journal, 1983, vol. 33, no. 5, pp. 15–20. 17. Disney M., Burt A., Wilkes P., Armston J., Duncanson L. New 3D Measurements of Large Redwood Trees for Biomass and Structure. Scientific Reports, 2020, no. 10, art. no. 16721. 18. Douhovnikoff V., Dodd R.S. Intra-Clonal Variation and a Similarity Threshold for Identification of Clones: Application to Salix exigua Using AFLP Molecular Markers. Theoretical and Applied Genetics, 2003, no. 106, pp. 1307–1315. https://doi.org/10.1007/s00122-003-1200-9 19. Eckert Christopher G. The Loss of Sex in Clonal Plants. Ecology and Evolutionary Biology of Clonal Plants: Proceedings of Clone-2000. An International Workshop Held in Obergurgl. Springer Netherlands, 2002, pp. 279–298. 20. 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