vir-nwТруды по прикладной ботанике, генетике и селекцииProceedings on applied botany, genetics and breeding2227-88342619-0982N.I. Vavilov All-Russian Institute of Plant Genetic Resources10.30901/2227-8834-2022-3-194-203vir-nw-1358Research ArticleИММУНИТЕТ КУЛЬТУРНЫХ РАСТЕНИЙ И ИХ ДИКИХ РОДИЧЕЙIMMUNITY OF CULTIVATED PLANTS AND THEIR WILD RELATIVESSolanum nigrum L. – резерватор вироида веретеновидности клубней картофеляSolanum nigrum L. is a potato spindle tuber viroid reservoirhttps://orcid.org/0000-0003-3383-2973МироненкоН. В.MironenkoN. V.

Мироненко Нина Васильевна - доктор биологических наук, ведущий научный сотрудник.

196608, Санкт-Петербург, Пушкин, шоссе Подбельского, 3.

Nina V. Mironenko - Dr. Sci. (Biology), Leading Researcher, All-Russian Research Institute of Plant Protection.

3 Podbelskogo Hwy., Pushkin, St. Petersburg 196608.

nina2601mir@mail.ruhttps://orcid.org/0000-0003-1479-7746ХюттиА. В.KhiuttiA. V.

Хютти Александр Валерьевич - кандидат биологических наук, старший научный сотрудник.

196608, Санкт-Петербург, Пушкин, шоссе Подбельского, 3.

Alexander V. Khiutti - Cand. Sci. (Biology), Senior Researcher, All-Russian Research Institute of Plant Protection.

3 Podbelskogo Hwy., Pushkin, St. Petersburg 196608.

alexanderkhyutti@mail.ruhttps://orcid.org/0000-0002-1016-2148КыроваЕ. И.KyrovaE. I.

Кырова Елена Игоревна - аспирант, младший научный сотрудник.

196608, Санкт-Петербург, Пушкин, шоссе Подбельского, 3.

Elena I. Kyrova - Postgraduate Student, Associate Researcher, All-Russian Research Institute of Plant Protection.

3 Podbelskogo Hwy., Pushkin, St. Petersburg 196608.

blairkot@gmail.comhttps://orcid.org/0000-0003-1419-7134ЛашинаН. М.LashinaN. M.

Лашина Нина Михайловна - кандидат биологических наук, старший научный сотрудник.

196608, Санкт-Петербург, Пушкин, шоссе Подбельского, 3.

Nina M. Lashina - Cand. Sci. (Biology), Senior Researcher, All-Russian Research Institute of Plant Protection.

3 Podbelskogo Hwy., Pushkin, St. Petersburg 196608.

nlashina@mail.ruhttps://orcid.org/0000-0001-7368-0797АфанасенкоО. С.AfanasenkoO. S.

Афанасенко Ольга Сильвестровна - доктор биологических наук, профессор РАН, ведущий научный сотрудник.

196608, Санкт-Петербург, Пушкин, шоссе Подбельского, 3.

Olga S. Afanasenko - Dr. Sci. (Biology), Professor of the RAS, Leading Researcher, All-Russian Research Institute of Plant Protection.

3 Podbelskogo Hwy., Pushkin, St. Petersburg 196608.

olga.s.afan@gmail.comВсероссийский научно-исследовательский институт защиты растенийРоссияAll­-Russian Research Institute of Plant ProtectionRussian Federation2022041020221833194203Copyright © Мироненко Н.В., Хютти А.В., Кырова Е.И., Лашина Н.М., Афанасенко О.С., 20222022Мироненко Н.В., Хютти А.В., Кырова Е.И., Лашина Н.М., Афанасенко О.С.Mironenko N.V., Khiutti A.V., Kyrova E.I., Lashina N.M., Afanasenko O.S.Данная работа распространяется под лицензией Creative Commons Attribution 4.0.This work is licensed under a Creative Commons Attribution 4.0 License.https://elpub.vir.nw.ru/jour/article/view/1358Актуальность

Актуальность. Среди сорных растений семейства пасленовых, потенциальных резерваторов инфекции вироида веретеновидности клубней картофеля (ВВКК), паслен черный (Solanum nigrum L.) повсеместно распространен на территории России.

Материалы и методы

Материалы и методы. Молекулярную диагностику вироида проводили методом ПЦР с обратной транскрипцией с использованием праймеров P3/P4, специфичных для ВВКК, и 6Pospi F/R, специфичных для семейства вироидов Pospiviroidae. Продукты амплификации клонировали и секвенировали, нуклеотидные последовательности депонировали в базе данных GenBank.

Результаты

Результаты. Методом ОТ-ПЦР показано, что 80–100% растений, собранных в Нижнем Новгороде и Тульской области, были инфицированы ВВКК. кДНК-ампликоны двух растений из Нижнего Новгорода (Sn1 и Sn2) и одного растения из Тульской области (Sn3) клонировали, по три клона каждого, и секвенировали. Анализ нуклеотидной изменчивости клонов выявил существенные генетические различия между штаммами ВВКК, при этом связь с географическим происхождением штаммов не наблюдается. Мутации U120C и U192C относительно референсного штамма VP35 (LC523658) обнаружены у всех 9 клонов. Изолят Sn1 близок к «картофельному» изоляту VP35 и отличается от него тремя мутациями. Изолят Sn2 представлен тремя идентичными клонами и отличается от VP35 девятью мутациями, а Sn3 имеет 22 мутации в области 49–310 позиции нуклеотидов суммарно для трех клонов.

После искусственного заражения здоровых растений S. nigrum в условиях теплицы подтвержден факт наличия ВВКК в инфицированных растениях и доказано, что вироид передается семенами растениям следующей генерации.

Заключение

Заключение. Впервые во внешне здоровых сорных растениях S. nigrum выявлен вироид веретеновидности клубней картофеля – ВВКК. Доказано, что вироид сохраняется в семенах S. nigrum и передается потомству. Обнаружена генетическая гетерогенность штаммов ВВКК, обитающих в растениях S. nigrum. Две мутации U120C и U192C были общими для всех тестированных клонов изолятов ВВКК, обнаруженных в S. nigrum.

Background

Background. Among the weeds of the Solanaceae family, a potential viroid infection reservoir, black nightshade (Solanum nigrum L.), is ubiquitous in Russia.

Materials and methods

Materials and methods. Molecular diagnostics of the viroid was performed using RT-PCR with the P3/P4 primers specific for PSTVd and 6Pospi F/R specific for the Pospiviroidae family of viroids. The amplification products were cloned and sequenced, and the nucleotide sequences were deposited in the GenBank database.

Results

Results. RT-PCR showed that 80–100% of plants were infected with PSTVd. The cDNA amplicons of two plants from Nizhny Novgorod (Sn1 and Sn2) and one plant from Tula Province (Sn3) were cloned (3 clones each) and sequenced. An analysis of the nucleotide variability in the clones revealed significant genetic differences between the PSTVd strains, independent of the geographical origin. Mutations U120C and U192C, compared with the reference strain VP35 (LC523658), were found in all 9 clones. The Sn1 isolate was close to the “potato” VP35 isolate and differed from it only by three mutations. The Sn2 isolate was represented by three identical clones and differed from VP35 by nine mutations, while Sn3 had 22 mutations in the region of 49–310 nucleotide positions in total for three clones. After artificial infection of healthy S. nigrum plants in a greenhouse, the presence of PSTVd in infected plants was confirmed and it was proved that the viroid was transmitted by seeds to plants of the next generation.

Conclusion

Conclusion. For the first time in externally healthy plants of S. nigrum, widely distributed in the potato fields of Russia (Nizhny Novgorod and Tula Province), the potato spindle tuber viroid, PSTVd, was detected. It was proved that the viroid was preserved in the seeds of S. nigrum and transmitted to their progeny. Genetic heterogeneity of PSTVd strains from S. nigrum plants was found. Two mutations, U120C and U192C, were common for all tested clones of PSTVd strains.

паслен черныйВВККОТ-ПЦРсеквенированиемутацииnightshade blackPSTVdRT-PCRsequencingmutationsРабота выполнена при финансовой поддержке гранта РНФ № 20-46-07001 «Молекулярно-генетические механизмы взаимоотношений в патосистеме <i>Solanum</i> spp. – вироид веретеновидности клубней картофеля (PSTVd) и особенности эпидемиологии вироида». Авторы благодарят рецензентов за их вклад в экспертную оценку этой работы.The research was supported by a grant from the Russian Science Foundation, No. 20-46-07001 “Molecular genetic mechanisms of relationships in the pathosystem of <i>Solanum</i> spp. – Potato spindle tuber viroid (PSTVd) and features of the viroid epidemiology”. The authors thank the reviewers for their contribution to the peer review of this work.ReferencesAdkar-Purushothama C.R., Bolduc F., Bru P., Perreault J.P. Insights into potato spindle tuber viroid quasi-species from infection to disease. Frontiers in Microbiology. 2020;11:1235. 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The authors declare that there are no conflicts of interest present.