Virus Detection in Phalaenopsis Leaves

Tissue blotting to reveal virus infection and distribution in inoculated Phalaenopsis leaves was performed as described previously [35 (link),42 (link)]. In brief, leaf slices from inoculated and adjacent non-inoculated tissues were printed onto Hybond™-N⁺ membranes (GE Healthcare Life Sciences, Buckinghamshire, UK) and hybridized with riboprobes specific to CymMV/ORSV full-length CP genes by using the DIG Nucleic Acid Detection Kit (Sigma-Aldrich, St. Louis, MO, USA). Northern blot analysis was performed, as described previously [43 (link),44 ], by using DIG-labeled probes targeted to the CP and 3′ UTR sequence of the pUCy1 and pUOS4 genomes.