RUDN Journal of Agronomy and Animal IndustriesRUDN Journal of Agronomy and Animal IndustriesВестник Российского университета дружбы народов. Серия: Агрономия и животноводство2312-797X2312-7988Peoples’ Friendship University of Russia named after Patrice Lumumba1956310.22363/2312-797X-2020-15-2-150-158Plant protectionЗащита растенийResearch ArticleA new primer set for amplification of ITS-rDNA in Ditylenchus destructorНовый набор праймеров для специфичной амплификации ITS-локусов рДНК Ditylenchus destructorMahmoudiNiloufarМахмудиНилоуфар

PhD candidate, Department of Agro-Biotechnology, Agrarian and Technological Institute

аспирант агробиотехнологического департамента, Аграрнo-технологический институт

niloofarmahmoodi@ymail.comNejadDavoud K.НеджадДавуд Картули

Assistant Professor of Forestry, Faculty of Desert Studies

доцент кафедры лесоводства, факультет исследований пустынь

Kartooli58@gmail.comShayanmehrFatemehШаянмерФатиме

PhD in Forestry, Natural Resources Faculty

кандидат наук, факультет природных ресурсов

niloofarmahmoodi@ymail.comPeoples’ Friendship University of RussiaРоссийский университет дружбы народовAll-Russian Plant Quarantine CentreВсероссийский центр карантина растенийSemnan UniversityСемнанский университетTarbiat Modares UniversityУниверситет Тарбиат Модарес15122020152VOL 15, NO2 (2020)ТОМ 15, №2 (2020)15015813062020Copyright ©; 2020, Mahmoudi N., Nejad D.K., Shayanmehr F.Copyright ©; 2020, Махмуди Н., Неджад Д.К., Шаянмер Ф.2020Mahmoudi N., Nejad D.K., Shayanmehr F.Махмуди Н., Неджад Д.К., Шаянмер Ф.http://creativecommons.org/licenses/by/4.0https://agrojournal.rudn.ru/agronomy/article/view/19563

A technique was developed for the identification of Ditylenchus destructor nematode belonging to the Ditylenchus genus, based on the use of different primers for polymerase chain reaction (PCR). Two universal ribosomal primers were amplified to the internal transcribed spacer region ITS-rDNA. The sequencing of PCR products confirmed the polymorphism between species. The primers were sensitive to generate a particular band of the correct size (300bp) from the DNA template of a single, separate D. destructor stage of development. Screening populations of D. destructor from Iran and the Russian Federation have tested the reliability of the primers, and the expected size of the band was produced for all test populations. Ditylenchus destructor closely related species have also been tested and no specific band was amplified. Such results showed that the primers currently developed are useful for quantifying the D. destructo r density in potato tuber.

Разработана методика идентификации нематоды Ditylenchus destructor , основанная на использовании различных праймеров для проведения полимеразной цепной реакции (ПЦР). Два универсальных рибосомных праймера были амплифицированы во внутренней транскрибируемой области спейсера ITS-рДНК. Секвенирование продуктов ПЦР подтвердило полиморфизм между видами. Праймеры достаточно чувствительны, чтобы генерировать определенную полосу правильного размера (300 п.н.) из ДНК-матрицы определенной стадии развития D. destructor . Скринингом популяций D. destructor из Ирана и Российской Федерации проверена надежность праймеров, и ожидаемый размер полосы был получен для всех тестируемых популяций. Близкородственные виды Ditylenchus destructor также были протестированы, и специфической имплифицированной полосы не было выявлено. Результаты исследований показали, что данные разработанные праймеры могут использоваться для количественной оценки плотности D. destructor в клубнях картофеля.

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