A calmodulin-like protein suppresses RNA silencing and promotes geminivirus infection by degrading SGS3 via the autophagy pathway in Nicotiana benthamiana
Fig 1
Interactions between NbCaM, NbSGS3 and NbRDR6.
(A and B) Yeast two-hybrid assays using NbCaM and NbSGS3 (A) or NbCaM and NbRDR6 (B). Serial 10-fold dilutions of yeast cells were made as indicated. Cells co-transformed with AD-T7-T+BD-T7-53 served as a positive control, and cells co-transformed with AD-T7-T+BD-T7-Lam, or with empty vectors pGBKT7 (BD) and pGADT7 (AD) served as negative controls. BD, GAL4 DNA binding domain; AD, GAL4 activation domain. (C and D) BiFC assays using NbCaM, NbSGS3 and NbRDR6 in H2B-RFP transgenic N. benthamiana leaves at 48 hours post infiltration. YFP fluorescence (green) was observed as a consequence of complementation of NbCaM and NbSGS3, or NbSGS3 and NbRDR6 tagged with 2YN and 2YC. Nuclei of tobacco leaf epidermal cells are indicated by expression of the H2B-RFP transgene (red). No YFP fluorescence was found between pairwise expression NbCaM and NbRDR6, or NbSGS3 and P3N-PIPO (D). Single channels from the merged image shown in (D) are shown in S2 Fig. Bars = 25 μm.