vavilovВавиловский журнал генетики и селекцииVavilov Journal of Genetics and Breeding2500-3259Institute of Cytology and Genetics of Siberian Branch of the RAS10.18699/VJ20.653vavilov-2774Research ArticleГЕНЕТИЧЕСКИЕ РЕСУРСЫ И БИОКОЛЛЕКЦИИPLANT GENETICSСравнительный анализ экспрессии генов у чайного растения (Camellia sinensis (L.) Kuntze) при низкотемпературном стрессеComparative analysis of gene expression in tea plant (Camellia sinensis (L.) Kuntze) under low-temperature stresshttps://orcid.org/0000-0002-0500-1198СамаринаЛ. С.SamarinaL. S.

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q11111w2006@yandex.ruМацькивА. О.MatskivA. O.

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КонинскаяН. Г.KoninskayaN. G.

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СимонянТ. А.SimonyanT. A.

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https://orcid.org/0000-0003-4213-8705МаляровскаяВ. И.MalyarovskayaV. I.

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МалюковаЛ. С.MalyukovaL. S.

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Федеральный исследовательский центр «Субтропический научный центр Российской академии наук»РоссияFederal Research Centre the Subtropical Scientific Centre of the Russian Academy of SciencesRussian Federation202027102020246598604Copyright © Самарина Л.С., Мацькив А.О., Конинская Н.Г., Симонян Т.А., Маляровская В.И., Малюкова Л.С., 20202020Самарина Л.С., Мацькив А.О., Конинская Н.Г., Симонян Т.А., Маляровская В.И., Малюкова Л.С.Samarina L.S., Matskiv A.O., Koninskaya N.G., Simonyan T.A., Malyarovskaya V.I., Malyukova L.S.This work is licensed under a Creative Commons Attribution 4.0 License.https://vavilov.elpub.ru/jour/article/view/2774

Низкотемпературный стресс – один из главных факторов, ограничивающих распространение и снижающих урожайность многих субтропических культур, в том числе и чая. Для эффективной селекции чая на устойчивость к морозу необходимо выявить генетические особенности ответа на холод у устойчивых генотипов и найти маркеры для определения доноров устойчивости в коллекциях. В настоящей работе проведен сравнительный анализ экспрессии 18 генов (ICE1, CBF1, DHN1, DHN2, DHN3, NAC17, NAC26, NAC30, bHLH7, bHLH43, P5CS, WRKY2, LOX1, LOX6, LOX7, SnRK1.1, SnRK1.2, SnRK1.3), вовлеченных в абиотический стрессовый ответ у двух контрастных по устойчивости генотипов чая в условиях холода и мороза. Низкотемпературный стресс индуцировали путем помещения растений в холодильные камеры и снижением температуры до 0…+2 °С на семь дней (холодовой стресс) с последующим снижением температуры до –4…–6 °С на пять дней (промораживание). Кондуктометрическим методом измеряли электропроводность тканей листа, в результате чего были подтверждены различия по признаку устойчивости у двух исследуемых генотипов чая: холодовое воздействие не приводило к изменению электропроводности тканей листа, но после промораживания этот показатель возрастал в большей степени у неустойчивого генотипа. Методом qRT-PCR анализировали относительный уровень экспрессии генов на фоне референсного гена актина. При индукции стресса показана повышенная экспрессия всех исследуемых генов. У устойчивого генотипа чая выявлен ряд генов, более активно экспрессирующихся по сравнению с неустойчивым генотипом: ICE1, CBF1, DHN2, NAC17, NAC26, bHLH43, WRKY2, P5CS, LOX6, SnRK1.1, SnRK1.3. Эти гены могут быть маркерами устойчивости для поиска доноров в коллекциях геноресурсов. Показано, что у устойчивого генотипа чая экспрессия генов холодового ответа начинается уже на стадии акклиматизации. Для дальнейших исследований комплексной устойчивости растений к низкотемпературному стрессу актуальным является изучение экспрессии этих генов в других органах чайного растения (побегах, корнях) при разной силе низкотемпературного воздействия.

Low-temperature stress is one of the main factors limiting the distribution and reducing the yield of many subtropical crops, including the tea crop. Efficient breeding to develop frost-tolerant cultivars requires a reliable set of genetic markers for identifying resistance donors, and that is why it is necessary to reveal the specific genetic response in frost-tolerant genotypes in comparison with frost- susceptible ones. In this work, we performed a comparative analysis of the expression of 18 tea genes (ICE1, CBF1, DHN1, DHN2, DHN3, NAC17, NAC26, NAC30, bHLH7, bHLH43, P5CS, WRKY2, LOX1, LOX6, LOX7, SnRK1.1, SnRK1.2, SnRK1.3) under cold and frost conditions in two tea genotypes, tolerant and susceptible. Low-temperature stress was induced by placing the potted plants in cold chambers and lowering the temperature to 0…+2 °С for 7 days (cold stress), followed by a decrease in temperature to –4…–6 °С for 5 days (frost stress). Relative electrical conductivity of leaf was measured in response to the stress treatments, and a significant difference in the frost tolerance of the two tea genotypes was confirmed. Cold exposure did not lead to a change in the electrical conductivity of leaf tissue. On the other hand, frost treatment resulted in increased REC in both genotypes and to a greater extent in the susceptible genotype. Increased expression of all the genes was shown during cold and frost. The genes that were strongly expressed in the tolerant tea genotype were revealed: ICE1, CBF1, DHN2, NAC17, NAC26, bHLH43, WRKY2, P5CS, LOX6, SnRK1.1, SnRK1.3. These genes can be proposed as markers for the selection of frost-tolerance donors in tea germplasm collections. Additionally, it was shown that the tolerant genotype is characterized by an earlier response to stress at the stage of cold acclimation. The study of the expression of the identified genes in different organs of tea plants and in different exposures to low temperature is relevant for further investigations.

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The authors declare that there are no conflicts of interest present.