Improved Double-Stranded RNA Extraction and Hyphal Tip-Touch Techniques for the Detection of Mycoviruses

• DOI: 10.1094/phytofr-08-23-0103-r
• Publication Date: 2023-10-13T19:07:21Z
• Authors: Samira Mokhtari; Akhtar Ali
• Keywords: Mycovirus; RNA Silencing

Mycoviruses infect a wide range of fungal species, and many them have double-stranded RNA (dsRNA) genomes. Using dsRNA technique makes it easier to detect unknown mycoviruses in fungi. Previously, few studies reported extraction methods from tissues, but the procedures required significant amount starting material time, particularly growing cultures. In this study, we report an efficient quick method that is reproducible extract quality small culture short time. Extracted was subsequently used Illumina sequencing reverse transcription-polymerase chain reaction assays, followed by Sanger sequencing, confirm identity mycovirus. Additionally, rapid named hyphal tip-touch (HTT) developed identify known without need for extraction. The HTT uses edge mycelia as template preparing complementary DNA transcription reaction, which polymerase assay sequencing. [Formula: see text] Copyright © 2024 Author(s). This open access article distributed under CC BY-NC-ND 4.0 International license .