Classification of virus-positive spontaneous plants was based on plant morphology and DNA barcode rbcL and matK genes, according to Fazekas et al. [33 (link)]. First, DNA extraction was done using CTAB method [34 ]. Then, the rbcL and matK genes were amplified with Taq DNA Polymerase, recombinant (Invitrogen, Carlsbad, CA, USA) using the sets of primers SI_For/SI_Rev and KIM 3F/KIM 1R [35 (link)] (Table S1 ).
All amplicons were visualized by electrophoresis in agarose gel stained with ethidium bromide (Invitrogen, Carlsbad, CA, USA). PCR products were excised from the gel, purified, and Sanger sequenced at Macrogen Inc. (Seoul, Republic of Korea). Sequences were assembled in Geneious Prime® 2022.1.1. and analyzed using the BOLD Identification System website [36 (link)].
All amplicons were visualized by electrophoresis in agarose gel stained with ethidium bromide (Invitrogen, Carlsbad, CA, USA). PCR products were excised from the gel, purified, and Sanger sequenced at Macrogen Inc. (Seoul, Republic of Korea). Sequences were assembled in Geneious Prime® 2022.1.1. and analyzed using the BOLD Identification System website [36 (link)].
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