Fig. 3: The 60 bp InDel in the WRKY34 promoter results in different expression patterns of WRKY34 under cold stress.

a WRKY34 gene structure in cultivated and wild tomatoes. The gray box represents 5’ UTR, the green box represents the coding sequence, the orange box represents the promoter and the black line in the promoter represents the 60 bp InDel region. b Schematic diagram of the reporter constructs with the full-length promoters of WRKY34 or different mutated WRKY34 promoters fused to the LUC reporter gene in the vector pGreenII 0800-LUC, while the internal control REN reporter gene was driven by the CaMV 35S promoter. pSlW34, full-length promoter of SlWRKY34; pShW34, full-length promoter of ShWRKY34; pSlW34^+60bp, SlWRKY34 full-length promoter inserts a 60 bp InDel from ShWRKY34 promoter; pShW34^mW-box, ShWRKY34 full-length promoter mutates W-box in the 60 bp InDel; pShW34^mGATA-box, ShWRKY34 full-length promoter mutates GATA-box in the 60 bp InDel; pSlW34^+30bp, SlWRKY34 full-length promoter inserts a 30 bp InDel from ShWRKY34 promoter. c LUC relative activities driven by different promoters were determined in transient transgenic tobacco plants 6 h after cold stress. The LUC relative activity driven by pSlW34 in transient expressed tobacco leaves under normal conditions was set to “1”. Data are presented as means ± SD of six biological replicates. The experiment was repeated three times with similar results. Different letters indicate significant differences (P < 0.05) according to one-way ANOVA with Duncan’s multiple range test. d The 60 bp InDel of 181 cultivated tomatoes, 74 cherry tomatoes, 58 currant tomatoes and 63 wild tomatoes. e The 60 bp InDel of 63 different wild tomatoes. f The foldchange of WRKY34 expression levels under cold stress in different tomato varieties. The horizontal line in the middle indicates mean. The experiment used three biological replicates in one experiment and was repeated twice with similar results. Source data are provided as a Source Data file.