Birch is one of the main deciduous forest-forming species in the European part of Russia, but the impact of stress (in particular, drought) greatly limits its distribution. In this regard, it seems relevant to identify resistance mechanisms in order to select promising genotypes for their further reproduction. The aim of this work is to identify drought-tolerant genotypes of European birch (B. pendula Roth.), pubescent birch (B. pubescens Ehrh.) аnd their hybrids. For the study, we have taken samples of birch at the age of 26 years, which remained viable after the droughts of 2010 and 2013. Birch leaves have been selected in the third decade of June 2019 to analyze drought resistance. Leaves of birch, selected during the period with optimal conditions of air temperature and precipitation, have been used as control ones. A modified CTAB method has been proposed for RNA isolation. We studied the expression of genes encoding proteins involved in the activation of cell defense pathways under the influence of abiotic stress (pal, PR-1, PR-10, lea8, DREB2) during the drought period (June 2019). A study of the expression of genes encoding proteins of metabolic pathways that are activated in response to abiotic stress (phenylpropanoid pathway) associated with the pathogenesis of proteins (PR1 and PR10), transcription factors (DREB2), and late embryogenesis proteins (LEA) has been made. As a result of the effects of drought, a significant increase in the expression of pal, PR-1, PR-10 and DREB2 genes has been detected in the analyzed samples. At the same time, changes in lea8 gene expression were detected for two out of ten genotypes. The largest increase in expression for all five genes is shown for birch samples 29-58 and 233. It indicates the development of adaptive mechanisms in these genotypes and can characterize them as the most stable. The studied genes can be recommended as markers for the analysis of stress resistance in various species of woody plants
Use of planting material of forest trees with improved hereditary characteristics is one of the ways to increase the productivity and biological stability of forest stands. It requires taking measures to develop and improve selection base using modern approaches and methods of genetics and biotechnology. A molecular genetics assessment of clone plants of aspen (Populus tremula L.) and white poplar (Populus alba L.) from a long-term in vitro collection (up to 24 years), planted in a greenhouse and field conditions (nursery), was carried out. SSR loci of the PTR series (PTR5, PTR7, PTR8, PTR12, PTR14) were used as DNA markers. Evaluation of clones' ploidy was carried out on the basis of the diagnosis of "loss of heterozygosity" (LOH) effect. Analysis of 5 microsatellite loci of the specimens showed their high intraclonal genotypic stability and homogeneity in vitro and ex vitro. For the first time, data on the results of a comparative determination of ploidy using karyological and microsatellite analysis were presented. Based on the results of the SSR analysis, it can be concluded that the structure of molecular markers is stable among the samples of one clone that are in long-term cultivation. The ratio of the representation (dose) of electrophoretic variants of PCR products serves as an indirect sign of determining ploidy, but for its reliable assessment it is necessary to study the number of loci that are three times larger than the main set of chromosomes. The specimen also requires information on the amplification coefficient of the markers under study. Thus, it is necessary to use both chromosomal and microsatellite analyzes for reliable assessment of intraclonal homogeneity of various specimens, the development of understanding of clone genotypes formation and determination of their ploidy
УДК 630*232.13 ИДЕНТИФИКАЦИЯ И ИССЛЕДОВАНИЕ ЭКСПРЕССИИ ГЕНОВ СТРЕССОУСТОЙЧИВОСТИ У ТОПОЛЕЙ С. Г. Ржевский 1 Т. А. Гродецкая 1 кандидат биологических наук П. М. Евлаков 1 доктор биологических наук Т. П. Федулова 1 1 -ФГБУ «Всероссийский научно-исследовательский институт лесной генетики, селекции и биотехнологии» г. Воронеж, Российская ФедерацияДанная работа посвящена оценке влияния холодового стресса на физиологические параметры и генетический аппарат селекционно-ценных сортов тополя. В ходе исследования проводилась оценка динамики распускания почек и определение экспрессии гена DREB2 у образцов различных сортов тополя, подвергнутых искусственному промораживанию. Методология постановки опыта включала моделирование холодового стресса при помощи низкотемпературной морозильной камеры, лабораторное отращивание контрольных и промороженных черенков в условиях искусственного освещения. В дальнейшем осуществлялась регистрация онтогенетической динамики распускания почек. При сравнении интенсивности отрастания опытных и контрольных образцов выявлено, что у некоторых сортов ('Ведуга', 'Болид') подвергнутые проморозке черенки демонстрируют более интенсивное распускание почек, в то время как другие генотипы ('Ивантеевский', 'ЭД-120', 'Китайский') проявили отставание различной степени в скорости распускания почек промороженных образов от контрольных. По результатам фенологических наблюдений были отобраны представители с контрастными показателями ('Сакрау', 'Э.c.-38' и 'Волосистоплодный') и подвергнуты дальнейшему изучению на молекулярно-генетическом уровне. Анализ показал повышение экспрессии гена DREB2 у двух опытных образцов сортов тополя ('Волосистоплодный' и 'Сакрау') после воздействия отрицательных температур относительно контрольных образцов. Промораживание опытных растений тополя 'Э.c.-38' демонстрировало обратный результат. В целом можно заключить, что наблюдение динамики распускания почек и оценка экспрессии генов является достаточно информативным маркером для изучения влияния холодового стресса, однако для уточнения связи фенологических параметров с экспрессией генов стрессоустойчивости требуются дополнительные исследования.
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