Fig. 1

Identification of transcription factors FvCtf1α in the regulation of FvCUT1 and FvCUT4 in F. verticillioides. a Self activation activity was verificated by yeast two-hybrid assays. Yeast transformants harboring the BD-FvCtf1α/pGADT7 vectors were assayed for growth on selective plates (SD/-Leu/Trp/His/Ade), and X-α-Gal added to test for β-galactosidase (LacZ) activities. X-α-Gal: 5-Bromo-4-chloro-3-indolyl-α-D-galactoside. The pGBKT7-53/pGADT7-T and pGBKT7-Lam/pGADT7-T were used as positive and negative controls, respectively. Scale bars: 10 µm. b The localizations of CTF transcription factor FvCtf1α was observed in the nucleus with the nucleus stained by DAPI. c–e The expression patterns of three cutinase genes within 12 h of cutin induction: the expression levels of cutinase genes FvCUT1, FvCUT3, and FvCUT4 in WT and mutants were detected at 1, 2, 3, 4, 5, and 12 h, respectively. f FvCtf1α can bind the promoters of the two cutinase genes FvCUT1 and FvCUT4 by yeast one-hybrid. pAbAi::FvCUT1pro vector, pAbAi::FvCUT4pro vector, and pGADT7-FvCtf1α were constructed and the Y1H-Gold strain was sequentially transferred into two vectors, first transferred into the pAbAi::FvCUT1pro vector (or pAbAi::FvCUT4pro vector), selecting with SD/-Ura medium, and re-introduced into the pGADT7-FvCtf1α vector, selecting with SD/-Ura media with 0, 100, 150, 200 ng/mL Aureobasidin A (AbA).The vector pair pAbAi::p53pro/p53-pGADT7 transformants serves as the positive control and the vector pAbAi::p53pro/pGADT7 transformants serves as the negative control. g FvCtf1α can bind the promoters of the two cutinase genes FvCUT1 and FvCUT4 at GC-rich site by Multiple EM for Motif Elicitation (MEME) software. p < 0.01. h The expression of FvCUT1, FvCUT3, FvCUT4 were assayed by qRT-PCR under low H₂O₂ stress. i The expression level of three genes including β-oxidation (FvPOT1) and peroxisome biogenesis (FvPEX5, FvECL1) were compared by WT and ΔFvctf1α, respectively. The transcription level of the target gene was determined using qRT-PCR assay and calculated using the 2^−ΔΔCt method with TUB2 as reference gene. Analysis of variance is three independent repeated experiments and asterisks represent a significant difference. (t-test, *: p < 0.05, **: p < 0.01)