Fig 9.

Evidence for generation of nonhomologous recombinants exclusively from (+)-strand inoculum templates. (A) Features of B3ΔTLS agroconstructs designed to ectopically express (+)- or (−)-strand transcripts. Features of each construct are the same as those described in the legend of Fig. 1A. (B) Northern blot analysis of progeny RNA in infiltrated (top panel) and uninoculated systemic (bottom panel) leaves of N. benthamiana leaves infiltrated with the indicated mixture of agrocultures. Conditions of Northern blot hybridization with riboprobes are as described in the Fig. 1B legend. Approximately 10 μg of total RNA was subjected to Northern blot hybridization, and 1 μg of B123 was used as a marker for a positive control. The positions of the four WT BMV RNAs are shown to the left of each panel. Since nonhomologous recombinants generated from B3ΔTLS (+) strands are distinct in sequences from those shown in Fig. 3, they are referred to as B3R-γ. The positions of recombinant progeny generated from infiltrating B3ΔTLS (+) strands (B3R-γ and sgB4R-γ) and (−) strands (B3R-α and sgB4R-α) are shown to the right. (C) Profiles of recombinants generated during restoration of the deleted TLS region from B3. All features of recombinant clones are as described in the legend of Fig. 3.