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Updated: Aug 16, 2025
Combining Analysis of DNA in a Crude Virion Extraction with the Analysis of RNA from Infected Leaves to Discover New Virus Genomes
Published on: July 27, 2018
A thermal cycler-based, homogenization-free plant total nucleic acid extraction method for plant viruses and viroids
Shuichi Uchiyama1, Yuya Imamura2, Takayuki Matsuura3
1Narita Sub-station, Yokohama Plant Protection Station, 2159, Aza Tennamino, Komaino, Narita 282-0021, Japan.
Abstract:
In this study, we report a plant total nucleic acid (TNA) extraction method for nucleic acid (NA)-based assays of plant viruses and viroids. This method combines NA release by incubating sliced plant tissue in solution and NA purification using silica spin column. The method is performed using a thermal cycler and microcentrifuge and does not involve tissue homogenization. For a wide range of plant species, TNA can be extracted from petioles, midribs, and stems, in 30 min. PCR/RT-PCR assays using extracts from this method detected all three DNA viruses, 14 RNA viruses, and 4 viroids tested and plant internal controls were also available. This method does not involve hazardous chemicals, is cost-effective. The method is readily implemented in various laboratories, a simple, rapid, and labor-saving option for NA-based assays of plant viruses and viroids.
