Figure 3.

Effects of P Protein Phosphorylation on Full-Length BYSMV Transcription and Replication in N. benthamiana Leaves.

(A) Illustration of BYSMV genome organization and the binary Agrobacterium vectors harboring pBYG-P^WT, pBYG-P^S5A, pBYG-P^S5D, pGD-NLP^WT, and pGD-NLP^S5A.

(B) GFP foci in N. benthamiana leaves at 10 dpi with Agrobacterium cultures containing the indicated plasmids. GFP fluorescence was observed and photographed with a fluorescence microscope. Bar = 0.5 mm.

(C) Relative replication and transcription levels of recombinant BYSMV^WT and BYSMV^S5A as quantified by RT-qPCR. BYSMV^WT was rescued by coinfiltration of pGD-NLP^WT and pBYG-P^WT, while BYSMV^S5A was rescued by coinfiltration of pGD-NLP^S5A and pBYG-P^S5A. The values of viral replication and transcription supported by BYSMV^WT were set to 1.

(D) Relative replication and transcription levels of recombinant BYSMV^S5A and BYSMV^S5D as quantified by RT-qPCR. BYSMV^S5A and BYSMV^S5D were rescued by coinfiltration of pBYG-P^S5A and pBYG-P^S5D under the support of pGD-NLP^S5A. The values of viral replication and transcription supported by BYSMV^S5A were set to 1.

In (C) and (D), error bars indicate sd. Data points above the columns are the mean values of three independent experiments. Statistically significant differences were determined by Student’s t test. *P-value < 0.05; **P-value < 0.01.