A thermal cycler-based, homogenization-free plant total nucleic acid extraction method for plant viruses and viroids assay

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.