Supplementary Fig.S1. Map of Specific Primers Designed to Detect the Tobacco Ringspot Virus

Supplementary Fig.S1. Map of specific primers designed to detect the Tobacco ringspot virus

Supplementary Fig.S2.First selection of specific RT-PCR primer sets for theTobacco ringspot virus

Lane M, 100 bp step DNA Ladder maker (Genepia, Korea); lanes, specific primer set number of TRSV RT-PCR primer set; dot, first selectived RT-PCR primer sets.

Supplementary Fig.S3. RT-PCR amplification of the non-specific selection among the related reference viruses

Lane M, 100 bp step DNA Ladder maker (Genepia); lane TR, Tobacco ringspot virus; lane Ar, Arabis mosaic virus; lane CL, Cherry leaf roll virus; lane GF, Grapevine fanleaf virus; lane Rp, Raspberry ringspot virus; lane TB, Tomato black ring virus; lane To, Tomato ringspot virus; dot, second selectived RT-PCR primer sets.

Supplementary Fig.S4. RT-PCR amplification of the non-specific selection among the host related viruses

Lane M, 100 bp step DNA Ladder maker (Genepia); lane +, Tobacco ringspot virus; lane BB, Broad bean wilt virus; lane BC, Bean common mosaic virus; lane LM, Lettuce mosaic virus; lane MN, Melon necrotic spot virus; lane PS, Potato virus S; lane PY, Potato virus Y; lane TM, Tobacco mosaic virus; lane TG, Tobacco mild green mosaic virus; lane TN, Tobacco necrosis virus; lane TR, Tobacco rattle virus; lane TS, Tobacco streak virus; dot, third selectived RT-PCR primer sets.

Supplementary Fig.S5. RT-PCR amplification of the non-specific selection among the healthy crops

Lane M, 100 bp step DNA Ladder maker (Genepia); lane 1・5・9・13, potato; lane 2・6・10・14, melon; lane 3・7・11・15, lettuce; lane 4・8・12・16, soybean; lane +, Tobacco ringspot virus; -, negative control (double-distilled water).

Supplementary Fig.S6. Sensitivity test of the RT-PCR primer sets and selection of the nested PCR primer sets for the Tobacco ringspot virus detection

(A) Sensitivity test of the RT-PCR primer sets. Lane M, 100 bp step DNA Ladder maker (Genepia); other lane name, dulution of TRSV plasmid value(100-10-8). (B) Selection of the nested PCR primer sets. Lane M, 100 bp step DNA Ladder maker (Genepia); lane 1, TRSVN12/TRSC40 (415 bp); lane 2, TRSVN12/TRSVC90 (415 bp); lane 3, TRSVN20/TRSC40 (254 bp); lane 4, TRSVN20/TRSVC90 (254 bp); lane 5, TRSN40/TRSVC80 (194 bp); lane 6, TRSVN40/TRSVC70 (234 bp).

Supplementary Fig.S7. Restriction enzyme (XhoI) to cut the modified-positive plasmid

(A) Result of electrophoresis for restriction enzyme (Xho I). Lane M, 100 bp DNA Ladder maker; lane 1, TRSV nested PCR product for modified-positive plasmid (TRSVN12/TRSVC90); lane 2, result of restriction enzyme XhoI treatment. (B) Sequence of the modified-positive control plasmid into the restriction enzyme (Xho I) site.