Figure 1.

The DNA‐B genome of the tomato leaf curl New Delhi virus (ToLCNDV)‐OM isolate is required for mechanical transmissibility. (a) Physical maps of DNA infectious clones (pCB2A, pCB2B, pOM2A, and pOM2B) carrying the duplicated DNA‐A or DNA‐B genome of the ToLCNDV‐CB or ToLCNDV‐OM isolate. Genes encoded by DNA‐A or DNA‐B and the location of the intergenic region (IR) sequence are also indicated. The arrow indicates the direction of the transcription of each gene. (b) Images of Nicotiana benthamiana (10 days post‐inoculation, dpi), oriental melon, and cucumber plants (12 dpi) after mechanical inoculation with the wild‐type virus (pCB2A + pCB2B and pOM2A + pOM2B) or pseudorecombinant viruses (pCB2A + pOM2B and pOM2A + pCB2B) developing viral symptoms (O) or exhibiting no symptoms (X). The viral inoculum used for mechanical sap inoculation was prepared from diseased N. benthamiana after agroinoculation with an appropriate combination of the clones. (c) PCR detection of virus accumulation in the leaves after mechanical inoculation with pCB2A + pCB2B (lanes 1, 5, and 9), pOM2A + pOM2B (lanes 2, 6, and 10), pCB2A + pOM2B (lanes 3, 7, and 11), and pOM2A + pCB2B (lanes 4, 8, and 12). The specific DNA fragments amplified by PCR are indicated on the left. The pseudoinfectious clones leading to successful mechanical sap inoculation are indicated by asterisks