izvestiiatimacadИзвестия Тимирязевской сельскохозяйственной академииIZVESTIYA OF TIMIRYAZEV AGRICULTURAL ACADEMY0021-342XФГБОУ ВО РГАУ-МСХА имени К.А. Тимирязева10.26897/0021-342X-2026-2-84-104izvestiiatimacad-1103Research ArticleЗЕМЛЕДЕЛИЕ, РАСТЕНИЕВОДСТВО, ЗАЩИТА РАСТЕНИЙAGRONOMY, CROP PRODUCTION, PLANT PROTECTIONОценка применимости методов ПЦР для диагностики бактериального рака томатаApplicability assessment of PCR methods for the diagnosis of bacterial canker in tomatohttps://orcid.org/0000-0002-3084-0591ПисареваИ. Н.PisarevaI. N.

Ирина Николаевна Писарева, научный сотрудник научно-методического отдела бактериологии

140150, Московская обл., м.о. Раменский, р.п. Быково, ул. Пограничная, 32

Irina N. Pisareva, Research Associate at the Research and Methodology Department of Bacteriology

32 Pogranichnaya St., Bykovo, Ramensky Municipal District, Moscow Region, 140150

iruru@yandex.ruhttps://orcid.org/0000-0002-8564-8142БелошапкинаО. О.BeloshapkinaO. O.

Ольга Олеговна Белошапкина, д-р с.-х. наук, профессор, профессор кафедры защиты растений

127434, г. Москва, ул. Тимирязевская, 49

Olga O. Beloshapkina, DSc (Ag), Professor, Professor at the Department of Plant Protection

49 Timiryazevskaya St., Moscow, 127434

beloshapkina@rgau-msha.ruФГБУ «Всероссийский центр карантина растений» (ФГБУ «ВНИИКР»)РоссияAll-Russian Plant Quarantine CenterRussian FederationРоссийский государственный аграрный университет – МСХА имени К.А. ТимирязеваРоссияRussian State Agrarian University – Moscow Timiryazev Agricultural AcademyRussian Federation2026210520260284104Copyright © Писарева И.Н., Белошапкина О.О., 20262026Писарева И.Н., Белошапкина О.О.Pisareva I.N., Beloshapkina O.O.Данная работа распространяется под лицензией Creative Commons Attribution 4.0.This work is licensed under a Creative Commons Attribution 4.0 License.https://izvestiia.timacad.ru/jour/article/view/1103

Бактериальный рак томата – вредоносное заболевание, распространенное на культуре томата. Особую вредоносность возбудитель болезни Сlavibacter michiganensis (Smith; Davis et al.) Li et al. проявляет в защищенном грунте. Быстрое и точное выявление С. michiganensis в семенах томата при помощи современных методов диагностики позволит снизить риск распространения заболевания, повысить урожайность и качество продукции. Исследования выполнены в лаборатории научно-методического отдела бактериологии ФГБУ «ВНИИКР» в 2022-2023 гг. Цель исследований – оценка применимости трех ПЦР-тестов, предложенных в международных протоколах для диагностики возбудителя бактериального рака томата. В задачи работы входило определение основных критериев эффективности (аналитическая чувствительность и специфичность, селективность, воспроизводимость, повторяемость) праймерных систем. В работе использовались отечественные реактивы, что позволило снизить затраты и повысить доступность диагностики. ПЦР в соответствии с рекомендациями Sudarshana et al. и Bach et al. обладали высокой аналитической чувствительностью – 103 КОЕ/мл, а в соответствии с рекомендациями Oosterhof и Berendsen она составила 104 КОЕ/мл. Все праймерные системы показали 100%-ную специфичность без перекрестных реакций с другими штаммами. Селективность не выявлена. Повторяемость и воспроизводимость всех тестов составили 100% на уровне порога чувствительности. В результате оценки применимости трех праймерных систем метод ПЦР по Sudarshana et al. выбран как скрининговый для выявления бактериального рака томата в аналитических пробах из семян и вегетативных частей томата. Остальные ПЦР-тесты рекомендованы как подтверждающие тесты.

Bacterial canker of tomato is a harmful disease widespread in tomato crops. The pathogen Clavibacter michiganensis (Smith; Davis et al.) Li et al. exhibits particular virulence in protected ground conditions. Rapid and accurate detection of C. michiganensis in tomato seeds using modern diagnostic methods can help reduce the risk of disease spread and improve crop yield and product quality. The study was conducted at the Laboratory of the Research and Methodology Department of Bacteriology, All-Russian Plant Quarantine Center, in 2022-2023. The study aimed to assess the applicability of three PCR tests proposed in international protocols for diagnosing the bacterial canker in tomato. The objectives included determining the key performance criteria of primer systems: analytical sensitivity and specificity, selectivity, reproducibility, and repeatability. Domestic reagents were used in the study, which helped reduce costs and increase the accessibility of diagnostics. According to the protocols by Sudarshana et al. and Bach et al. the PCR tests demonstrated high analytical sensitivity of of 10³ CFU/mL, while according to the protocol by Oosterhof and Berendsen sensitivity was 10⁴ CFU/mL. All primer systems demonstrated 100% specificity, with no cross-reactions observed with other strains. No selectivity was detected. The repeatability and reproducibility of all tests reached 100% at the sensitivity threshold level. Based on the assessment of the three primer systems, the PCR method described by Sudarshana et al. was selected as a screening tool for detecting bacterial tomato canker in analytical samples from seeds and vegetative parts of the plant. The remaining PCR tests are recommended as confirmatory assays.

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The authors declare that there are no conflicts of interest present.