Figure 2.

Schematic representation of the viral vectors used

Viral vectors for delivery of sgPDS3-tFT, a single guide RNA (sgRNA) construct that comprises of a protospacer (blue diamond) targeting both N. benthamiana PDS homeologs, a conserved Cas9 scaffold (gray box) and a truncated A. thaliana Flowering locus T (tFT; a pink box).

(A) Diagram of the PVX vector system.¹ An Agrobacterium strain hosts pLX-PVX, a T-DNA vector with the pBBR1 origin and kanamycin resistance for PVX agroinoculation (see Figure S1).

(B) Diagram of the JoinTRV system.² An Agrobacterium strain simultaneously hosts pLX-TRV1 and pLX-TRV2, two compatible T-DNA vectors for TRV agroinoculation. pLX-TRV1 is a derivative of pLX-Z4, which includes the RK2 origin and gentamicin resistance²⁰; pLX-TRV2 is a derivative of pLX-B2, which includes the pBBR1 origin and kanamycin resistance.²⁰ RdRp, RNA-dependent RNA polymerase; TGB, triple gene block; CP, coat protein; RdRAP, RNA-dependent replication-associated protein; MP, movement protein; 16K, 16 kDa protein. Black arrows show the heterologous bamboo mosaic virus (BaMV) and pea early browning virus (PEBV) coat protein promoters; promoters, sgRNA and tFT, not at scale.