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<article article-type="research-article" dtd-version="1.3" xmlns:mml="http://www.w3.org/1998/Math/MathML" xmlns:xlink="http://www.w3.org/1999/xlink" xmlns:xsi="http://www.w3.org/2001/XMLSchema-instance" xml:lang="ru"><front><journal-meta><journal-id journal-id-type="publisher-id">vavilov</journal-id><journal-title-group><journal-title xml:lang="ru">Вавиловский журнал генетики и селекции</journal-title><trans-title-group xml:lang="en"><trans-title>Vavilov Journal of Genetics and Breeding</trans-title></trans-title-group></journal-title-group><issn pub-type="epub">2500-3259</issn><publisher><publisher-name>Institute of Cytology and Genetics of Siberian Branch of the RAS</publisher-name></publisher></journal-meta><article-meta><article-id pub-id-type="doi">10.18699/VJ19.460</article-id><article-id custom-type="elpub" pub-id-type="custom">vavilov-1867</article-id><article-categories><subj-group subj-group-type="heading"><subject>Research Article</subject></subj-group><subj-group subj-group-type="section-heading" xml:lang="ru"><subject>АКТУАЛЬНЫЕ ТЕХНОЛОГИИ ГЕНЕТИКИ РАСТЕНИЙ</subject></subj-group><subj-group subj-group-type="section-heading" xml:lang="en"><subject>MAINSTREAM TECHNOLOGIES IN PLANT GENETICS</subject></subj-group></article-categories><title-group><article-title>Простой и эффективный метод экстракции полярных метаболитов из листьев гуара (Cyamopsis tetragonoloba (L.) Taub.) для GС-МS метаболомного анализа</article-title><trans-title-group xml:lang="en"><trans-title>A simple and efficient method to extract polar metabolites from guar leaves (Cyamopsis tetragonoloba (L.) Taub.) for GC-MS metabolome analysis</trans-title></trans-title-group></title-group><contrib-group><contrib contrib-type="author" corresp="yes"><contrib-id contrib-id-type="orcid">https://orcid.org/0000-0002-5624-4245</contrib-id><name-alternatives><name name-style="eastern" xml:lang="ru"><surname>Теплякова</surname><given-names>С. Б.</given-names></name><name name-style="western" xml:lang="en"><surname>Teplyakova</surname><given-names>S. B.</given-names></name></name-alternatives><bio xml:lang="ru"><p>Санкт-Петербург</p></bio><bio xml:lang="en"><p>St. Petersburg</p></bio><email xlink:type="simple">Serafima.teplyakova@mail.ru</email><xref ref-type="aff" rid="aff-1"/></contrib><contrib contrib-type="author" corresp="yes"><contrib-id contrib-id-type="orcid">https://orcid.org/0000-0003-1778-2814</contrib-id><name-alternatives><name name-style="eastern" xml:lang="ru"><surname>Шаварда</surname><given-names>А. Л.</given-names></name><name name-style="western" xml:lang="en"><surname>Shavarda</surname><given-names>A. L.</given-names></name></name-alternatives><bio xml:lang="ru"><p>Санкт-Петербург</p></bio><bio xml:lang="en"><p>St. Petersburg</p></bio><xref ref-type="aff" rid="aff-2"/></contrib><contrib contrib-type="author" corresp="yes"><contrib-id contrib-id-type="orcid">https://orcid.org/0000-0003-3992-5353</contrib-id><name-alternatives><name name-style="eastern" xml:lang="ru"><surname>Шеленга</surname><given-names>Т. В.</given-names></name><name name-style="western" xml:lang="en"><surname>Shelenga</surname><given-names>T. V.</given-names></name></name-alternatives><bio xml:lang="ru"><p>Санкт-Петербург</p></bio><bio xml:lang="en"><p>St. Petersburg</p></bio><xref ref-type="aff" rid="aff-1"/></contrib><contrib contrib-type="author" corresp="yes"><contrib-id contrib-id-type="orcid">https://orcid.org/0000-0003-4576-1527</contrib-id><name-alternatives><name name-style="eastern" xml:lang="ru"><surname>Дзюбенко</surname><given-names>Е. А.</given-names></name><name name-style="western" xml:lang="en"><surname>Dzyubenko</surname><given-names>E. A.</given-names></name></name-alternatives><bio xml:lang="ru"><p>Санкт-Петербург</p></bio><bio xml:lang="en"><p>St. Petersburg</p></bio><xref ref-type="aff" rid="aff-1"/></contrib><contrib contrib-type="author" corresp="yes"><contrib-id contrib-id-type="orcid">https://orcid.org/0000-0002-2578-6279</contrib-id><name-alternatives><name name-style="eastern" xml:lang="ru"><surname>Потокина</surname><given-names>Е. К.</given-names></name><name name-style="western" xml:lang="en"><surname>Potokina</surname><given-names>E. K.</given-names></name></name-alternatives><bio xml:lang="ru"><p>Санкт-Петербург</p></bio><bio xml:lang="en"><p>St. Petersburg</p></bio><xref ref-type="aff" rid="aff-3"/></contrib></contrib-group><aff-alternatives id="aff-1"><aff xml:lang="ru">Федеральный исследовательский центр Всероссийский институт генетических ресурсов растений им. Н.И. Вавилова (ВИР)<country>Россия</country></aff><aff xml:lang="en">Federal Research Center the N.I. Vavilov All-Russian Institute of Plant Genetic Resources (VIR)<country>Russian Federation</country></aff></aff-alternatives><aff-alternatives id="aff-2"><aff xml:lang="ru">Санкт-Петербургский государственный университет;&#13;
Ботанический институт им. В.Л. Комарова Российской академии наук<country>Россия</country></aff><aff xml:lang="en">St. Petersburg State University;&#13;
Komarov Botanical Institute, RAS<country>Russian Federation</country></aff></aff-alternatives><aff-alternatives id="aff-3"><aff xml:lang="ru">Федеральный исследовательский центр Всероссийский институт генетических ресурсов растений им. Н.И. Вавилова (ВИР);&#13;
Санкт-Петербургский государственный университет<country>Россия</country></aff><aff xml:lang="en">Federal Research Center the N.I. Vavilov All-Russian Institute of Plant Genetic Resources (VIR);&#13;
St. Petersburg State University<country>Russian Federation</country></aff></aff-alternatives><pub-date pub-type="collection"><year>2019</year></pub-date><pub-date pub-type="epub"><day>25</day><month>02</month><year>2019</year></pub-date><volume>23</volume><issue>1</issue><fpage>49</fpage><lpage>54</lpage><permissions><copyright-statement>Copyright &amp;#x00A9; Теплякова С.Б., Шаварда А.Л., Шеленга Т.В., Дзюбенко Е.А., Потокина Е.К., 2019</copyright-statement><copyright-year>2019</copyright-year><copyright-holder xml:lang="ru">Теплякова С.Б., Шаварда А.Л., Шеленга Т.В., Дзюбенко Е.А., Потокина Е.К.</copyright-holder><copyright-holder xml:lang="en">Teplyakova S.B., Shavarda A.L., Shelenga T.V., Dzyubenko E.A., Potokina E.K.</copyright-holder><license license-type="creative-commons-attribution" xlink:href="https://creativecommons.org/licenses/by/4.0/" xlink:type="simple"><license-p>This work is licensed under a Creative Commons Attribution 4.0 License.</license-p></license></permissions><self-uri xlink:href="https://vavilov.elpub.ru/jour/article/view/1867">https://vavilov.elpub.ru/jour/article/view/1867</self-uri><abstract><p>Гуар Cyamopsis tetragonoloba (L.) Taub. – новая для России сельскохозяйственная культура, востребованная в газо-, нефтедобывающей и пищевой промышленности. В связи с развитием «омиксных» технологий и для выявления ценных для селекции генов представляет интерес сравнительное изучение различных сортов и линий гуара с помощью метаболомики и функциональной геномики. Для массового скрининга метаболомных профилей образцов гуара из коллекции Всероссийского института генетических ресурсов растений им. Н.И. Вавилова с использованием GС-МS (Gas Chromatography-Mass Spectrometry) метаболомного анализа на первых этапах важно подобрать наиболее оптимальный метод экстракции метаболитов из анализируемых образцов. Результатом метаболомного анализа чаще всего является статистическая модель различий в корреляционной структуре метаболитной сети изучаемых объектов. Надежное квантирование метаболитных профилей критично для различения сортов одной культуры, так как профили метаболитов в тканях листа у растений одного вида, культивируемых в равных условиях, практически не отличаются по набору метаболитов. В метаболомной практике при подготовке образцов к GС-MS-анализу распространено два способа экстракции полярных соединений. Один из широко используемых методов пробоподготовки основан на длительной экстракции метаболитов из цельных незамороженных тканей с помощью растворителя метанола, а другой – на краткосрочной метанольной экстракции метаболитов из замороженного и подвергнутого гомогенизации материала. Преимущества и недостатки этих двух методов побудили нас к разработке нового подхода, позволяющего избежать затруднений при анализе метаболомных профилей листьев различных сортов гуара. Предложенный нами метод объединяет преимущества двух выше указанных способов пробоподготовки, а именно: исключает потерю метаболитов на этапе центрифугирования и способствует полной деструкции всех клеточных стенок, обеспечивая максимальный уровень экстракции полярных метаболитов. Метод состоит в том, что лист быстро замораживается в жидком азоте с последующим размораживанием в холодном метаноле. При этом ткани листа сохраняют морфологическую целостность, и последующее центрифугирование, необходимое при гомогенизации, исключается. Нами была показана эффективность использования этого усовершенствованного метода на образцах листьев трех линий гуара. Установлено, что количество экстрагируемых метаболитов увеличивается более чем в пять раз по сравнению с метанольной экстракцией из свежего листа без замораживания и более чем в два раза в сравнении с экстракцией метанолом после замораживания и гомогенизации. Экстракция метаболитов новым методом позволяет проводить GС-MS-анализ образцов гуара с наименьшими потерями и высокой точностью, необходимой при выявлении сортовых различий.</p></abstract><trans-abstract xml:lang="en"><p>Guar (Cyamopsis tetragonoloba (L.) Taub.) is an agricultural crop species new to Russia and is in demand by the gas, oil and food industries. Due to the progress of “omics” technologies and the marker-assisted selection, there is a huge interest in the studies that compare the metabolites of various guar varieties, employing metabolomics as a method of functional genomics. For a large-scale screening of guar germplasm from the VIR collection, it is important to choose an efficient method to extract metabolites from samples. The accuracy of the assessment of the content of metabolites in samples is crucial for distinguishing varieties within the crop, since the metabolome profiles of plants within the same species differ mainly in the quantitative ratio of metabolites, and not in their qualitative composition. In metabolome practice, two methods of extracting polar compounds are usually employed in the preparation of samples for GC-MS analysis. One of the widely used methods of sample preparation is the long-term extraction of metabolites from whole tissues with the aid of a methanol solvent. Another method of sample preparation is based on the short-term methanol extraction of metabolites from frozen and homogenized material. The advantages and disadvantages of these two methods revealed in the course of our work have prompted us to develop a new approach that avoids some difficulties in analyzing the metabolic profiles of leaves of various guar varieties. The method we suggested combines the advantages of the two above-mentioned approaches of sample preparation, namely eliminates the loss of metabolites due to centrifugation and ensures the complete destruction of all cell walls, ensuring the maximum extraction level of polar metabolites. The essence of the new method is that the leaf is rapidly frozen in liquid nitrogen with subsequent thawing in cold methanol. Thus, leaf tissues retain morphological integrity, and subsequent centrifugation, necessary for homogenization, is skipped. We have checked the effectiveness of this improved method by experiments with leaf samples of three guar genotypes. It has been shown that the amount of extracted metabolites increases more than 5-fold compared to extraction with methanol from fresh unfrozen leaf tissues and more than 2-fold compared to extraction with methanol after freezing and homogenization. Extraction of metabolites using the new method allows the GC-MS analysis of guar samples to be conducted with the least loss and high accuracy required to distinguish varieties.</p></trans-abstract><kwd-group xml:lang="ru"><kwd>Cyamopsis tetragonoloba (L.) Taub.</kwd><kwd>гуар</kwd><kwd>газовая хроматография</kwd><kwd>масс-спектрометрия</kwd><kwd>метаболомика</kwd><kwd>экстракция метаболитов</kwd></kwd-group><kwd-group xml:lang="en"><kwd>Cyamopsis tetragonoloba (L.) Taub</kwd><kwd>guar</kwd><kwd>gas chromatography</kwd><kwd>mass spectrometry</kwd><kwd>metabolomics</kwd><kwd>metabolite extraction</kwd></kwd-group></article-meta></front><back><ref-list><title>References</title><ref id="cit1"><label>1</label><citation-alternatives><mixed-citation xml:lang="ru">Лоскутов И.Г., Шеленга Т.В., Конарев А.В., Шаварда А.Л., Блинова Е.В., Дзюбенко Н.И. Метаболомный подход к сравнительному анализу диких и культурных видов овса (Avena L.). Вавиловский журнал генетики и селекции. 2016;20(5):636-642. 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