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. 2021 Apr 1;10(4):679. doi: 10.3390/plants10040679

Figure 3.

Figure 3

Repression of the TwTPS27a and TwTPS27b promoters by transient overexpression of TwMYC2a and TwMYC2b in tobacco leaves. (A) Sketch map of the reporter and effector vectors for GUS staining and GUS fluorimetric assays. 27aP::GUS and 27bP::GUS represent the promoter expression vectors constructed by introducing the promoter sequences of TwTPS27a (1496 bp) and TwTPS27b (1862 bp) into the pBI121 vector, respectively. Based on the GUS staining and GUS fluorimetric analysis of the infiltrated tobacco leaves, TwMYC2a and TwMYC2b significantly decreased the promoter activity of TwTPS27a (B) and TwTPS27b (C). Different combinations of the mixed bacterial suspension were infiltrated into the left and right sides of the same tobacco leaves, respectively. 27aP and 27bP represent the 27aP::GUS and 27bP::GUS. M2a and M2b represent the 35S::TwMYC2a and 35S::TwMYC2b. GUS activity of the infiltrated leaves was measured after infiltration for 72 h. The value of GUS activity was measured as pmol of 4-MU generated per hour per μg protein. All data presented here are means ± SD of three biological replicates. Statistical significance was determined by Student’s t-test (** p < 0.01). These assays were repeated three times with similar results.