*4.6. Subcellular Localization Assay*

The ORFs of *EXO70-V* genes (without stop codon) were amplified from the *pTOPO-Blunt* Vector, then inserted into the *pCambia1305-GFP* vector, which contains a green fluorescent protein (GFP) reporter gene driven by the CaMV 35S promoter, using homologous cloning technology as per the manufacturer's instructions (Vazyme, Nanjing, China) (Table S3). Then it was introduced into *Agrobacterium tumefaciens* (strain GV3101) bacteria by a freeze–thaw procedure and grown in Luria-Bertani (LB) medium at 28 ◦C for 2 or 3 d.

*Agrobacterium tumefaciens* (strain GV3101) bacteria containing fusion constructed were grown in Luria-Bertani (LB) medium with both rifampicin and kanamycin (0.05 µg/mL) at 28 ◦C overnight. The bacterial cells were centrifuged and resuspended in an infiltration solution (10 mM MES pH 5.6, 0.1 mM Acetosyringone, 10 mM MgCl2) to a final OD<sup>600</sup> = 1.5. Bacterial suspensions were infiltrated into five- to six-week growing stage leaves of *N. benthamiana* by depressing the plunger of a 1-mL disposable needleless syringe into the abaxial side of leaves [85,86]. The fluorescence signals were observed 48–60 h after injection and images were captured using a confocal laser scanning microscope (LSM780; Carl Zeiss, Jena, Germany) according to the methods described by Wang et al. [87].
