МОЖЛИВОСТІ ВИКОРИСТАННЯ МЕТОДУ ЗТ-ПЛР ДЛЯ МОНІТОРИНГУ ВІРУСНОГО ІНФЕКЦІЙНОГО ПРОЦЕСУ У РОСЛИН

• Дата: 2025
• Автори: Щукін, Ю. С., Коваленко, О. Г.
• Формат: Стаття
• Мова: Українська
• Опубліковано: Institute of Agrocultural Microbiology and Agro-industrial Manufacture of NAAS of Ukraine 2025
• Теми: віруси рослин; моніторинг вірусного інфекційного процесу у рослин; вірус тютюнової мозаїки (Tobacco mosaic virus; TMV); вірус смугастої мозаїки пшениці (Wheat Streak Mosaic Virus; WSMV); полімеразна ланцюгова реакція в реальному часі зі стадією зворотної транскрипції (ЗТ-ПЛР); екстракція нуклеїнових кислот з рослинного матеріалу
• Онлайн доступ: https://smic.in.ua/index.php/journal/article/view/551
• Назва журналу: Agriciltural microbiology

Objective. To determine the possibilities of using the RT-PCR method for monitoring viral infectious processes in plants. Methods. Extraction of nucleic acids from tobacco and wheat plants using the STAV protocol and real-time polymerase chain reaction with reverse transcription stage (RT-PCR). Results. A real-time PCR method for monitoring viral processes in plants has been created and tested. The study presents optimized protocols for isolation, primers, and fluorescent probes, as well as buffer composition and a selected amplification protocol. The data obtained indicate the sensitivity, specificity, and linearity of the real-time PCR test systems, which allows them to be optimized for quantitative determination for further use in monitoring the viral infection process in plants. Conclusions. The example of TMV and WSMV viruses demonstrates the fundamental possibility of using RT-PCR systems for quantitative assessment of the viral infection course in plants. The results obtained can be used to study other DNA- and RNA-containing viruses. The proposed method has obvious advantages over the methods of detection and monitoring of viral infections in plants (sensitivity, specificity, the possibility of quantitative determination of virus-specific products) that are currently used. However, the peculiarity of its practical implementation is the need for expensive equipment and reagents, as well as the availability of trained, qualified personnel. The practical value of the method will increase after its adaptation to the monitoring of economically important viruses of agricultural crops that reproduce vegetatively (potatoes, strawberries, fruit crops).