1. (A) RdRP 5‘UTR3‘UTR CP MP SP6 (B) (C) 3 dpi 15 dpi (D) Supplementary Figure 1 Infectivity analysis of Nicotiana tabacum. Schematic illustration of the TMV RNA expression vector pT2SB (a) and the infection construct of this vector (b). Leaves of Nicotiana tabacum cv. Xanthi NN (c) and Xanthi nn (d) were dusted with carborundum and rub inoculated with RNA either generated by in vitro transcription with pT2SB as template (left) or purified from wtTMV (right). After infection, plants are grown under standard conditions. dpi, days post infection.

2. pT2SB:SP1-1 Nicotiana tabacum cv. Xanthi NN Nicotiana tabacum cv. Xanthi nn Supplementary Figure 2 Infectivity analysis of Nicotiana tabacum. (a) Schematic illustration of the infection construct of pT2SB:SP1-1. (b) Nicotiana tabacum cv. Xanthi NN and Xanthi nn were infected with RNA derived from in vitro transcription of pT2SB:SP1-1. Both the resistant and the susceptible cultivar show distinct HR-lesions 6 dpi that restricted the recombinant TMV from local cell-to-cell movement in the leaf and inhibited formation of infection. RdRP 5‘UTR3‘UTR CP - M - AMP MP SP6 (A) (B)

3. Supplementary Figure 3 pAGRO::T2SB-CP_cc_SP1-1cc as example for an agroinfiltration vector. RdRP, RNA-dependent RNA polymerase; p35S, CaMV 35S promoter; Ap R, ampicillin resistance gene; UTR, untranslated region; CNBr, cyanogen bromide; RB, right border; LB, left border; Nos term, nopaline synthase terminator. pAGRO:T2SB-CP_cc_SP1-1 12130 bp

4. 55 43 34 26 17 1 2 M kDa Supplementary Figure 4 Acetic acid extraction of T2SB_CP_cc_SP1-1. The acid extract was dialysed against water oN and proteins pI precipitated by adjusting the pH to 5.5 with NaOH. Precipitated proteins were centrifugated and resuspended in 1/5 volume of buffer corresponding to the volume of the acetic acid extract and separated on 15%-SDS-PAGE. 1, T2SB_CP_cc_SP1-1; 2, wtTMV CP as size control. Relative molecular marker standards are shown on the right. M, marker; kDa, kilodalton.