vuzbiochemiИзвестия вузов. Прикладная химия и биотехнологияProceedings of Universities. Applied Chemistry and Biotechnology2227-29252500-1558ИРНИТУ10.21285/2227-2925-2023-13-4-483-493RSMMRTvuzbiochemi-1118Research ArticleФИЗИКО-ХИМИЧЕСКАЯ БИОЛОГИЯPHYSICOCHEMICAL BIOLOGYВлияние состава питательной среды на сохранение жизнеспособности и генетической стабильности эксплантов хризантемы садовой при депонировании in vitroEffect of growth medium composition on the viability and genetic stability of Chrysanthemum × morifolium Ramat. explants under in vitro cold storage conditionshttps://orcid.org/0000-0001-7628-9646ИвановаН. Н.IvanovaN. N.

Иванова Наталия Николаевна, к.б.н., старший научный сотрудник

298648, Республика Крым, г. Ялта, ул. Никитский спуск, 52

Natalya N. Ivanova, Cand. Sci. (Biology), Senior Researcher

52, Nikitskiy Spusk, Yalta, 298648, Crimea Republic

nnivanova2017@yandex.ruhttps://orcid.org/0000-0003-3853-0210ЦюпкаВ. А.TsiupkaV. A.

Цюпка Валентина Анатольевна, к.б.н., старший научный сотрудник

298648, Республика Крым, г. Ялта, ул. Никитский спуск, 52

Valentina A. Tsiupka, Cand. Sci. (Biology), Senior Researcher

52, Nikitskiy Spusk, Yalta, 298648, Crimea Republic

valentina.brailko@yandex.ruhttps://orcid.org/0000-0001-5220-5700КорзинаН. В.KorzinaN. V.

Корзина Наталья Васильевна, к.б.н., заведующий лабораторией

298648, Республика Крым, г. Ялта, ул. Никитский спуск, 52

Natalia V. Korzina, Cand. Sci. (Biology), Head of the Laboratory

52, Nikitskiy Spusk, Yalta, 298648, Crimea Republic

natali.korz@yandex.ruФГБУН «Ордена Трудового Красного Знамени Никитский ботанический cад – Национальный научный центр РАН»РоссияOrder of the Red Banner Nikita Botanical Gardens – National Scientific Center RASRussian Federation202326122023134483493Copyright © Иванова Н.Н., Цюпка В.А., Корзина Н.В., 20232023Иванова Н.Н., Цюпка В.А., Корзина Н.В.Ivanova N.N., Tsiupka V.A., Korzina N.V.Данная работа распространяется под лицензией Creative Commons Attribution 4.0.This work is licensed under a Creative Commons Attribution 4.0 License.https://vuzbiochemi.elpub.ru/jour/article/view/1118

Для сохранения in vitro ценных сортов хризантемы садовой перспективно культивирование растений при низких положительных температурах. Целью данного исследования было изучение влияния концентраций хлорхолинхлорида в специализированной среде на жизнеспособность, физиологические и морфо-анатомические показатели растений хризантемы сортов Excel, Sheer Purple, William Seward, а также их генетическую стабильность после 12 месяцев депонирования в условиях генобанка in vitro. Верхушки микропобегов in vitro без листьев помещали на среду 1/4 Мурасиге – Скуга + 60,0 г/л сахарозы и 0,2–0,8 г/л хлорхолинхлорида. Экспланты сохраняли в холодильных камерах при температуре 4 °С. После 12 месяцев сохранения наблюдали снижение кинетики роста по мере увеличения концентрации хлорхолинхлорида в среде. Жизнеспособность эксплантов снижалась c 96 до 91% с ростом концентрации хлорхолинхлорида. Эффективный фотохимический квантовый выход фотосистемы II снизился более чем в два раза при депонировании по сравнению с контролем. Был выявлен ряд гистологических особенностей листьев депонируемых в генобанке in vitro эксплантов: у листовых пластинок уменьшались толщина листа и плотность мезофилла, деградировала палисадная ткань, отмечена высокая степень регуляции транспирации. С помощью SSR- и ISSR-анализа показано, что микропобеги исследуемых сортов хризантемы после депонирования in vitro идентичны исходным растениям. Проведенные исследования показали, что наличие в среде 0,6–0,8 г/л хлорхолинхлорида снижает кинетику роста эксплантов при сохранении высокой жизнеспособности и генетической стабильности.

Cultivation of plants at low positive temperatures is promising for the in vitro preservation of valuable Chrysanthemum × morifolium Ramat. varieties. The study aims to examine the effect of chlorocholine chloride concentrations in a specialized medium on the viability and the physiological and morpho-anatomical parameters of chrysanthemum plants (Excel, Sheer Purple, and William Seward varieties), as well as their genetic stability after 12 months of cold storage under in vitro gene bank conditions. In vitro microshoot tips (without leaves) were placed on a 1/4 Murashige and Skoog medium supplemented with 60.0 g/L sucrose and 0.2–0.8 g/L chlorocholine chloride. The explants were stored in cooling chambers at a temperature of 4 °C. After 12 months of preservation, a reduction was observed in growth kinetics with increasing concentration of chlorocholine chloride in the medium. The viability of explants decreased from 96 to 91% at higher concentrations of chlorocholine chloride. The effective photochemical quantum yield of photosystem II decreased more than twice under cold storage conditions as compared to the control. Several histological features were identified in the leaves of in vitro explants cold-stored in the gene bank: decreased in leaf thickness and mesophyll density in leaf blades, degradation of palisade tissue, and a high degree of transpiration regulation. The SSR and ISSR analyses were used to show that the microshoots of the examined chrysanthemum varieties were identical to the original plants following in vitro cold storage. The conducted studies revealed that the presence of 0.6–0.8 g/L chlorocholine chloride in the medium reduces the growth kinetics of explants while maintaining their high viability and genetic stability.

Chrysanthemum × morifolium Ramat.эксплантморфометрические параметрыдепонирование in vitroISSR-анализChrysanthemum × morifolium Ramat.explantmorphometric parametersin-vitro cold storageISSR analysisРабота выполнена в рамках госзадания Министерства образования и науки Российской Федерации FNNS-2022-0002. Исследования осуществлены на оборудовании уникальной научной установки «Научный центр биотехнологии, геномики и депонирования растений» («ФИТОБИОГЕН») ФГБУН «Ордена Трудового Красного Знамени Никитский ботанический сад – Национальный научный центр РАН» (г. Ялта, Российская Федерация).Ministry of Education and Science of the Russian Federation supported the work (State task FNNS-20220002). Unique equipment of the “Scientific Center for Biotechnology, Genomics and Plant Deposition” (FITOBIOGEN) of Nikita Botanical Gardens (Yalta, Russian Federation) was used for the research.ReferencesSarasan V., Cripps R., Ramsay M., Atherton C., McMichen M., Prendergast G., et al. 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The authors declare that there are no conflicts of interest present.