Botanica Pacifica. A journal of plant science and conservation Preprint
Article first published online: 19 MAY 2018 | DOI: 10.17581/bp.2018.07113
Flow cytometry analysis of the relative content of nuclear DNA in Nitraria schoberi L. seeds
Evgeny V. Banaev, Mariya A. Tomoshevich & Mariya S. Voronkova
Central Siberian Botanical Garden SB RAS, Novosibirsk, Russia
The effectiveness of using the flow cytometry method (FCM) to analyze the genome size of the genus Nitraria L. plants is discussed for the first time, a technique having been developed to study the relative nuclear DNA content in resting seeds of Nitraria schoberi L. The effect of adding polyvinylpyrrolidone antioxidant to the extracting buffer was studied to assess the presence of metabolites inhibiting nuclei staining with propidium iodide in leaves of a standard sample Raphanus sativus L. and N. schoberi seeds. The existence of inhibitors in leaves of the standard Raphanus sativus L. ‘Saxa’ was established. Intensity of fluorescence of nuclei isolated from N. schoberi seeds did not change under the antioxidant, indicating the absence of the staining inhibitor in the seeds. In addition, N. schoberi seeds are a convenient material for analysis of genome size, as they do not contain endosperm, are easily transported and easily stored.
Банаев Е.В., Томошевич М.А., Воронкова М.С. Особенности анализа относительного содержания ядерной ДНК в семенах Nitraria schoberi L. методом проточной цитометрии. Впервые проведена оценка эффективности использования метода проточной цитометрии (FCM) для анализа размера генома растений рода Nitraria L., разработана методика исследования относительного содержания ядерной ДНК в покоящихся семенах Nitraria schoberi L. Для оценки присутствия метаболитов, ингибирующих окрашивание ядер иодидом пропидия, в листьях стандартного образца Raphanus sativus L. ‘Saxa’, и семенах N. choberi проведено изучение влияния добавления антиоксиданта поливинилпиролидона в экстрагирующий буфер. Установлено наличие ингибиторов в листьях стандартного образца Raphanus sativus L. ‘Saxa’. Интенсивность флюоресценции ядер, выделенных из семян N. schoberi, не меняется под действием антиоксиданта, что свидетельствует об отсутствии в семенах ингибиторов окрашивания. Кроме того, семена N. schoberi являются удобным материалом для анализа размера генома, поскольку не содержат эндосперм, легко транспортируются и хорошо хранятся.
Keywords: Nitraria schoberi, flow cytometry, genome size, relative DNA content, проточная цитометрия, размер генома, относительное содержание ДНК
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