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. 2022 Sep 21;190(4):2775–2796. doi: 10.1093/plphys/kiac441

Figure 2.

Figure 2

RIN-independent and ethylene- and auxin-dependent mode of action of SlERF.D7. A, Analysis of transactivation potential of SlERF.D7 in yeast by growing transformants on synthetic dextrose (SD) media-lacking leucine (Leu), tryptophan (Trp), histidine (His), and adenine (Ade). B, The presence of CArG Box, a putative fruit-specific element in addition to putative ethylene and AuxREs in the promoter of SlERF.D7 gene. The cis-acting regulatory elements identified are represented by different color boxes. C, RT-qPCR analysis of SlERF.D7 transcripts in total RNA samples extracted from WT MG fruit samples treated with 100 µM ethrel, 100 µM 1-MCP, 100 μM IAA, and 100 µM PCIB. Error bars, mean ± sd of three biological replicates. Asterisks indicate the statistical significance using Student’s t test: *, 0.01 < P-value < 0.05; **, 0.001 < P-value < 0.01, *** P-value < 0.001. E8, an ethylene response gene; SAUR68, an auxin response gene. D and E, Identification of DNA binding activity of RIN to the promoter of SlERF.D7 with (D) growth performance of transformants on SD/−Leu−/Trp/−His medium containing 50 mM 3-AT. Binding of RIN to the promoter of tomato ACC synthase2 (ACS2) gene-: positive control, binding of RIN to the promoter of tomato ACC oxidase1 (ACO1) gene-: negative control (E, in vivo interaction study of RIN to promoters of SlERF.D7, SlACS2, and SlACO1 via GUS reporter assays in N. benthamiana leaves.