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. 2014 Oct 22;16(5):435–448. doi: 10.1111/mpp.12201

Figure 7.

figure

The F‐box function of P0T u is required for RPO1 (Resistance to POleroviruses 1) function. (A) Transient expression of P0T u failed to elicit a cell death response in resistant Nicotiana glutinosa (left) when a mutation was introduced into the F‐box motif (P0LP1). The LP1 mutation abolishes P0 function as a suppressor of gene silencing in N. glutinosa, seen by the co‐expression of P0 and green fluorescent protein (GFP) in leaves of susceptible plants (right). (B) Three new P0T u F‐box mutants, designated L (L60A and L63A), P (P64G) and K (R57K and L60A), were cloned with a carboxy‐terminal haemagglutinin (HA) tag for detection on immunoblots. Agrobacterium carrying wild‐type and mutant P0T u constructs or empty pBIN61 vector was co‐infiltrated with pBIN61‐GFP and examined at up to 6 days post‐infiltration (dpi). Mutants were all impaired in both hypersensitive response (HR) elicitation on TW59 and silencing suppression when visualized under UV light at 6 dpi, but all P0T u proteins accumulated to similar levels when detected on immunoblots with anti‐HA horseradish peroxidase (HRP). The Coomassie brilliant blue‐stained blot is shown for comparison of protein loading.