FgFAD12-GFP fusion construct was generated using the ClonExpress® II One Step Cloning Kit (Vazyme, Nanjing, China). To generate the FAD12-GFP fusion construct, the FAD12 gene with its native promoter was amplified with primer pair Fad12/HF-HR (Table S1 ). Subsequently, the FAD12 gene fragments were cloned into the pKNT-GFP vector by Hind III/Kpn I double-digested to obtain FAD12-GFP fusion construct [36 (link)]. The resulting FgFAD12-GFP fusion construct was transformed into ∆Fgfad12 mutants. Transformants containing hygromycin and genecin resistance were verified by PCR, and GFP signals were observed with epifluorescence microscope (Nikon, DS-Ri1, Tokyo, Japan).
For co-localization transformants, FgKar2-mCherry-T-vector was obtained from Shuli Cao of Jiangsu Academy of Agricultural Sciences. FgKar2-mCherry and Fad12-GFP were co-transformed into the wild-type PH-1. Transformants were examined GFP and mCherry signals with epifluorescence microscope (Nikon, DS-Ri1, Tokyo, Japan).
For co-localization transformants, FgKar2-mCherry-T-vector was obtained from Shuli Cao of Jiangsu Academy of Agricultural Sciences. FgKar2-mCherry and Fad12-GFP were co-transformed into the wild-type PH-1. Transformants were examined GFP and mCherry signals with epifluorescence microscope (Nikon, DS-Ri1, Tokyo, Japan).
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