PbNAC71 was ubiquitinated by PbRNF217 and then degraded. A) PbRNF217 promotes the ubiquitination of PbNAC71 in N. benthamiana leaves. MYC-PbNAC71 was expressed alone or co-expressed with PbRNF217-GFP in N. benthamiana leaves. The proteins immunoprecipitated using the anti-MYC antibody were analyzed in a western blot involving anti-MYC and antiubiquitin antibodies. IP, immunoprecipitate; IB, immunoblot; Ubi, ubiquitin. B) The abundance of MYC-PbNAC71 in “Duli” roots was analyzed by immunoblotting using the anti-MYC antibody. OE, overexpressing. C) Degradation of PbNAC71 mediated by PbRNF217 in a cell-free system. Proteins were extracted from the PbRNF217-OE “Duli” roots or the wild-type “Duli” roots and then incubated with PbNAC71-GST and MG132 or DMSO for 0, 0.5, and 1.5 h at 22 °C. The abundance of PbNAC71 was determined via a western blot analysis involving the anti-GST antibody. β-Actin was used as the reference protein. WT, wild type.