<?xml version="1.0" encoding="UTF-8"?>
<!DOCTYPE article PUBLIC "-//NLM//DTD JATS (Z39.96) Journal Publishing DTD v1.3 20210610//EN" "JATS-journalpublishing1-3.dtd">
<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/VJ17.230</article-id><article-id custom-type="elpub" pub-id-type="custom">vavilov-911</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</subject></subj-group><subj-group subj-group-type="section-heading" xml:lang="en"><subject>Mainstream technologies</subject></subj-group></article-categories><title-group><article-title>Генетическая паспортизация картофеля на основе мультиплексного анализа 10 микросателлитных маркеров</article-title><trans-title-group xml:lang="en"><trans-title>Multiplexed set of 10 microsatellite markers for identification of potato varieties</trans-title></trans-title-group></title-group><contrib-group><contrib contrib-type="author" corresp="yes"><name-alternatives><name name-style="eastern" xml:lang="ru"><surname>Колобова</surname><given-names>О. С.</given-names></name><name name-style="western" xml:lang="en"><surname>Kolobova</surname><given-names>O. S.</given-names></name></name-alternatives><bio xml:lang="ru"><p>Москва</p></bio><bio xml:lang="en"><p>Moscow</p></bio><email xlink:type="simple">iab@iab.ac.ru</email><xref ref-type="aff" rid="aff-1"/></contrib><contrib contrib-type="author" corresp="yes"><name-alternatives><name name-style="eastern" xml:lang="ru"><surname>Малюченко</surname><given-names>О. П.</given-names></name><name name-style="western" xml:lang="en"><surname>Maluchenko</surname><given-names>O. P.</given-names></name></name-alternatives><bio xml:lang="ru"><p>Москва</p></bio><bio xml:lang="en"><p>Moscow</p></bio><xref ref-type="aff" rid="aff-1"/></contrib><contrib contrib-type="author" corresp="yes"><name-alternatives><name name-style="eastern" xml:lang="ru"><surname>Шалаева</surname><given-names>Т. В.</given-names></name><name name-style="western" xml:lang="en"><surname>Shalaeva</surname><given-names>T. V.</given-names></name></name-alternatives><bio xml:lang="ru"><p>Москва</p></bio><bio xml:lang="en"><p>Moscow</p></bio><xref ref-type="aff" rid="aff-1"/></contrib><contrib contrib-type="author" corresp="yes"><name-alternatives><name name-style="eastern" xml:lang="ru"><surname>Шанина</surname><given-names>Е. П.</given-names></name><name name-style="western" xml:lang="en"><surname>Shanina</surname><given-names>E. P.</given-names></name></name-alternatives><bio xml:lang="ru"><p>пос. Исток, Екатеринбург</p></bio><bio xml:lang="en"><p>Istok, Ekaterinburg</p></bio><xref ref-type="aff" rid="aff-2"/></contrib><contrib contrib-type="author" corresp="yes"><name-alternatives><name name-style="eastern" xml:lang="ru"><surname>Шилов</surname><given-names>И. А.</given-names></name><name name-style="western" xml:lang="en"><surname>Shilov</surname><given-names>I. A.</given-names></name></name-alternatives><bio xml:lang="ru"><p>Москва</p></bio><bio xml:lang="en"><p>Moscow</p></bio><xref ref-type="aff" rid="aff-1"/></contrib><contrib contrib-type="author" corresp="yes"><name-alternatives><name name-style="eastern" xml:lang="ru"><surname>Алексеев</surname><given-names>Я. И.</given-names></name><name name-style="western" xml:lang="en"><surname>Alekseev</surname><given-names>Ya. I.</given-names></name></name-alternatives><bio xml:lang="ru"><p>Москва</p></bio><bio xml:lang="en"><p>Moscow</p></bio><xref ref-type="aff" rid="aff-1"/></contrib><contrib contrib-type="author" corresp="yes"><name-alternatives><name name-style="eastern" xml:lang="ru"><surname>Велишаева</surname><given-names>Н. С.</given-names></name><name name-style="western" xml:lang="en"><surname>Velishaeva</surname><given-names>N. S.</given-names></name></name-alternatives><bio xml:lang="ru"><p>Москва</p></bio><bio xml:lang="en"><p>Moscow</p></bio><xref ref-type="aff" rid="aff-1"/></contrib></contrib-group><aff-alternatives id="aff-1"><aff xml:lang="ru">Федеральное государственное бюджетное научное учреждение «Всероссийский научно-исследовательский институт сельскохозяйственной биотехнологии»<country>Россия</country></aff><aff xml:lang="en">All-Russia Research Institute of Agricultural Biotechnology<country>Russian Federation</country></aff></aff-alternatives><aff-alternatives id="aff-2"><aff xml:lang="ru">Федеральное государственное бюджетное научное учреждение «Уральский научно-исследовательский институт сельского хозяйства»<country>Россия</country></aff><aff xml:lang="en">Ural Research Institute of Agricultural Science<country>Russian Federation</country></aff></aff-alternatives><pub-date pub-type="collection"><year>2017</year></pub-date><pub-date pub-type="epub"><day>24</day><month>03</month><year>2017</year></pub-date><volume>21</volume><issue>1</issue><fpage>124</fpage><lpage>127</lpage><permissions><copyright-statement>Copyright &amp;#x00A9; Колобова О.С., Малюченко О.П., Шалаева Т.В., Шанина Е.П., Шилов И.А., Алексеев Я.И., Велишаева Н.С., 2017</copyright-statement><copyright-year>2017</copyright-year><copyright-holder xml:lang="ru">Колобова О.С., Малюченко О.П., Шалаева Т.В., Шанина Е.П., Шилов И.А., Алексеев Я.И., Велишаева Н.С.</copyright-holder><copyright-holder xml:lang="en">Kolobova O.S., Maluchenko O.P., Shalaeva T.V., Shanina E.P., Shilov I.A., Alekseev Y.I., Velishaeva N.S.</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/911">https://vavilov.elpub.ru/jour/article/view/911</self-uri><abstract><p>Генетическая паспортизация сортов картофеля является востребованным инструментом для усовершенствования системы регистрации и сертификации, защиты прав селекционеров и контроля генетической однородности сортов. Наиболее перспективным подходом для различения и идентификации сортов на генетическом уровне продолжает оставаться использование микросателлитных маркеров на основе коротких тандемных повторов STR. Их амплификация с последующим электрофорезом высокого разрешения позволяет получить индивидуальную характеристику сорта – ДНК-профиль. При анализе большого количества образцов методика получения генетических профилей требует создания мультиплексных систем фрагментного анализа, что повышает требования как к отбору маркеров, так и к протоколу анализа. Для генотипирования сортов картофеля были отобраны 10 информативных STR-маркеров: STI0032, STG0016, STI0001, STI0004, STM1104, STM5127, STI0030, STI0033, STI0014, STM5114, проведен дизайн праймеров с разделением локусов по длинам и флуоресцентным меткам, разработан протокол выделения ДНК из клубней и других частей растений картофеля на основе CTAB-метода, оптимизирован состав реакционной смеси и условия ПЦР для эффективной мультиплексной амплификации и анализа локусов методом капиллярного электрофореза. Использование генетического анализатора позволяет определять длину аллелей с точностью до одного нуклеотида и получать оцифрованные генетические профили. В исследование были включены 41 сорт отечественной и зарубежной селекции и 26 селекционных образцов. Получены уникальные профили для каждого генотипа, позволяющие различать и идентифицировать сорта и сортообразцы, для 8 сортов проведена оценка однородности. Предложенная методика генетической паспортизации позволяет провести анализ большого количества образцов в формате 96-луночного планшета в короткие сроки.</p></abstract><trans-abstract xml:lang="en"><p>Genetic identification of potato varieties is a demanded instrument for development of new cultivars registration system, protection of plant breeders’ rights, and variety homogeneity control. The most perspective approach for distinction and identification of varieties continues to remain the use of short tandem repeats. STR amplification with the subsequent high resolution electrophoresis allows such a unique characteristic of a variety to be obtained as the DNA profile. A large scale of samples requires the creation of a robust and time-saving technique based on fragment sizing. We selected 10 polymorphic STR loci of potato: STI0032, STG0016, STI0001, STI0004, STM1104, STM5127, STI0030, STI0033, STI0014, STM5114 and designed a multiplex panel for potato DNA profiling. Fluorescent labelling of primers and size distinction of amplicones allowed us to use one tube for PCR and capillary electrophoresis. We also modified the CTAB-protocol for DNA extraction from tubers and other parts of potato plants, the PCR mix recipe and the amplification protocol for good results. Using Genetic Analyzer allows the length of alleles to be defined with an accuracy of one nucleotide and digitized genetic profiles to be developed. We created a unique DNA profile for each of 40 varieties and 23 breeding lines from Russia and other countries and evaluated the homogeneity of 8 varieties. The proposed technique оf potato DNA profiling allows a large number of samples to be rapidly analyzed in the 96-well plate format.</p></trans-abstract><kwd-group xml:lang="ru"><kwd>Solanum</kwd><kwd>картофель</kwd><kwd>фрагментный анализ</kwd><kwd>STR</kwd><kwd>генетическая идентификация</kwd></kwd-group><kwd-group xml:lang="en"><kwd>Solanum</kwd><kwd>potato</kwd><kwd>fragment analysis</kwd><kwd>STR</kwd><kwd>genetic identification</kwd></kwd-group><funding-group xml:lang="ru"><funding-statement>State Budgeted Project 05742015-0001 “Development of New Generation Technologies for Large-Scale Genetic Analysis of Crop Forms for Breeding Acceleration”</funding-statement></funding-group></article-meta></front><back><ref-list><title>References</title><ref id="cit1"><label>1</label><citation-alternatives><mixed-citation xml:lang="ru">Antonova O.Y., Shvachko N.A., Novikova L.Y., Shuvalov O.Y., Kostina L.I., Klimenko N.S., Shuvalova A.R., Gavrilenko T.A. Genetic diversity of potato varieties bred in Russia and near-abroad countries based on polymorphism of SSR-loci and markers associated with resistance R-genes. Vavilovskii Zhurnal Genetiki i Selektsii = Vavilov Journal of Genetics and Breeding. 2016;20(5):596-606. DOI 10.18699/VJ16.181. (in Russian)</mixed-citation><mixed-citation xml:lang="en">Antonova O.Y., Shvachko N.A., Novikova L.Y., Shuvalov O.Y., Kostina L.I., Klimenko N.S., Shuvalova A.R., Gavrilenko T.A. Genetic diversity of potato varieties bred in Russia and near-abroad countries based on polymorphism of SSR-loci and markers associated with resistance R-genes. Vavilovskii Zhurnal Genetiki i Selektsii = Vavilov Journal of Genetics and Breeding. 2016;20(5):596-606. DOI 10.18699/VJ16.181. (in Russian)</mixed-citation></citation-alternatives></ref><ref id="cit2"><label>2</label><citation-alternatives><mixed-citation xml:lang="ru">Doyle J.J., Doyle J.L. A rapid DNA isolation procedure for small quantities of fresh leaf tissue. Phytochem. Bull. 1987;19:11-15.</mixed-citation><mixed-citation xml:lang="en">Doyle J.J., Doyle J.L. A rapid DNA isolation procedure for small quantities of fresh leaf tissue. Phytochem. Bull. 1987;19:11-15.</mixed-citation></citation-alternatives></ref><ref id="cit3"><label>3</label><citation-alternatives><mixed-citation xml:lang="ru">Gishlan M., Nunez J., Herrera M., Pignataro J., Guzman F., Bonierbale M., Spooner D.M. Robust and highly informative microsatellite-based genetic identity kit for potato. Mol. Breeding. 2009;23:377-388. DOI 10.1007/s11032-008-9240-0.</mixed-citation><mixed-citation xml:lang="en">Gishlan M., Nunez J., Herrera M., Pignataro J., Guzman F., Bonierbale M., Spooner D.M. Robust and highly informative microsatellite-based genetic identity kit for potato. Mol. Breeding. 2009;23:377-388. DOI 10.1007/s11032-008-9240-0.</mixed-citation></citation-alternatives></ref><ref id="cit4"><label>4</label><citation-alternatives><mixed-citation xml:lang="ru">Gosudarstvennyy reestr selektsionnykh dostizheniy, dopushchennykh k ispol’zovaniyu. T. 1. Sorta rasteniy (ofitsial’noe izdanie) [State Register of Breeding Achievements Approved for Use. Varieties of Plants (official publication). V. 1. Plant cultivars]. Moscow: Rosinformagrotekh Publ., 2016; http://reestr.gossort.com/docs/reestr_2016.pdf. (in Russian)</mixed-citation><mixed-citation xml:lang="en">Gosudarstvennyy reestr selektsionnykh dostizheniy, dopushchennykh k ispol’zovaniyu. T. 1. Sorta rasteniy (ofitsial’noe izdanie) [State Register of Breeding Achievements Approved for Use. Varieties of Plants (official publication). V. 1. Plant cultivars]. Moscow: Rosinformagrotekh Publ., 2016; http://reestr.gossort.com/docs/reestr_2016.pdf. (in Russian)</mixed-citation></citation-alternatives></ref><ref id="cit5"><label>5</label><citation-alternatives><mixed-citation xml:lang="ru">Nei M. Analysis of gene diversity in subdivided populations. Proc. Natl. Acad. Sci. USA. 1973;70:3321-3323.</mixed-citation><mixed-citation xml:lang="en">Nei M. Analysis of gene diversity in subdivided populations. Proc. Natl. Acad. Sci. USA. 1973;70:3321-3323.</mixed-citation></citation-alternatives></ref><ref id="cit6"><label>6</label><citation-alternatives><mixed-citation xml:lang="ru">Reid A., Kerr E.M. A rapid simple sequence repeat (SSR)-based identification method for potato cultivars. Plant Genet. Resources. 2007; 5(1):713. DOI 10.1017/S1479262107192133.</mixed-citation><mixed-citation xml:lang="en">Reid A., Kerr E.M. A rapid simple sequence repeat (SSR)-based identification method for potato cultivars. Plant Genet. Resources. 2007; 5(1):713. DOI 10.1017/S1479262107192133.</mixed-citation></citation-alternatives></ref><ref id="cit7"><label>7</label><citation-alternatives><mixed-citation xml:lang="ru">Ryzhova N.N., Martirosyan E.V., Kochieva E.Z. Analysis of microsatellite locus polymorphism in potato (Solanum tuberosum) cultivars of Russian breeding. Genetika = Genetics (Moscow). 2010;46(4):425-430. (in Russian)</mixed-citation><mixed-citation xml:lang="en">Ryzhova N.N., Martirosyan E.V., Kochieva E.Z. Analysis of microsatellite locus polymorphism in potato (Solanum tuberosum) cultivars of Russian breeding. Genetika = Genetics (Moscow). 2010;46(4):425-430. (in Russian)</mixed-citation></citation-alternatives></ref><ref id="cit8"><label>8</label><citation-alternatives><mixed-citation xml:lang="ru">Velishaeva N.S., Khavkin E.E., Shilov L.A. Genotyping of potato and its wild relatives by microsatellite analysis. Doklady Rossiyskoy Akademii Selskokhozyaystvennykh Nauk = Proceedings of the Russian Academy of Agricultural Sciences. 2006;5:35. (in Russian)</mixed-citation><mixed-citation xml:lang="en">Velishaeva N.S., Khavkin E.E., Shilov L.A. Genotyping of potato and its wild relatives by microsatellite analysis. Doklady Rossiyskoy Akademii Selskokhozyaystvennykh Nauk = Proceedings of the Russian Academy of Agricultural Sciences. 2006;5:35. (in Russian)</mixed-citation></citation-alternatives></ref><ref id="cit9"><label>9</label><citation-alternatives><mixed-citation xml:lang="ru">Yessimseitova A.K., Shustov A.V., Ahmetollaev I.A., Krassavin V.F., Kakimzhanova A.A. Molecular-genetic certification of potato varieties and forms using SSR-markers. Eurasian J. Appl. Biotechnol. (Biotechnology: Theory and Practice). 2015;2. Available at: http://www.biotechlink.org/2-2015/article6. (in Russian)</mixed-citation><mixed-citation xml:lang="en">Yessimseitova A.K., Shustov A.V., Ahmetollaev I.A., Krassavin V.F., Kakimzhanova A.A. Molecular-genetic certification of potato varieties and forms using SSR-markers. Eurasian J. Appl. Biotechnol. (Biotechnology: Theory and Practice). 2015;2. Available at: http://www.biotechlink.org/2-2015/article6. (in Russian)</mixed-citation></citation-alternatives></ref></ref-list><fn-group><fn fn-type="conflict"><p>The authors declare that there are no conflicts of interest present.</p></fn></fn-group></back></article>
