*2.3. Subcellular Localization of CaHSP18.1a Protein*

The ORF (open reading frame) of *CaHSP18.1a* without a termination codon was PCRamplified using a specific primer pair (Supplementary Table S1). The resulting *CaHSP18.1a* fragment was cloned into the pVBG2307: GFP vector with *Xba*I and *Kpn*I restriction sites. The pVBG2307:*CaHSP18.1a*: GFP fusion protein transient expression vector and the control vector pVBG2307: GFP, after having been successfully constructed, were transformed into *Agrobacterium tumefaciens* strain GV3101, which was then injected into tobacco (*Nicotiana tabacum*) leaves to induce transient expression. After dark cultivation for approximately 36 h, epidermis samples of tobacco leaves were photographed under a fully automatic upright fluorescence microscope on the public platform of the College of Horticulture, Northwest A&F University, and the fluorescence patterns in the cells were observed; we specifically used the method described by Yu et al. [41].
