Fig. 6

Biological function analysis of PeNAC67, PeKAN2 and PeSCL23 in Phalaenopsis. a The fusion constructs of PeNAC67-Flag, PeMADS3-Myc, PeKAN2-HA tags were transformed into P.Big chill in different combinations, and Myc/Flag/HA antibody was used for immunoprecipitation. b The fusion constructs of PeSCL23-HA, PeMADS3-Flag and PeKAN2-Myc tags were transformed into P.Big chill in different combinations, and Myc/Flag/HA antibody was used for immunoprecipitation. c Representative phenotypic analysis for three independent PeNAC67 and PeKAN2 double silencing lines are presented. The red arrow represents the part where the mutation occurs after PeNAC67 and PeKAN2 VIGS treatment, and the blue arrow represents the corresponding part in the Mock. d and (e) Relative transcript levels of PeKAN2, PeNAC67 and PeSCL23 in Mock and VIGS-treated lines (d) and relative transcript levels of PeMADS3, PeMADS4 and PeMADS9 in Mock and VIGS-treated lines (e). f The epidermal cells of wild-type control and those of PeNAC67/PeKAN2 silenced flowers representative phenotypic analysis for three independent silencing lines was presented. Bar = 50 μm. g “Lip-like petal” mutant phenotype of P. equestris var.trilip. The red arrow represents the part where the mutation of “Lip-like petal”. h-j The expression of PeNAC67, PeKAN2 and PeSCL23 in “Lip-like petal” mutant phenotype of P. equestris var.trilip. (k) “Lip” mutant phenotype of P. equestris var.trilip. The red arrow represents the part where the mutation of “Lip petal”. l-n The expression of PeNAC67, PeKAN2 and PeSCL23 in lip mutant phenotype of P. equestris var.trilip. o-r In-situ localization of PeKAN2 (p), PeSCL23 (q) and PeNAC67 (r) transcripts in P. equestris flower buds. Longitudinal sections were hybridized with DIG-labeled antisense. o A negative control was performed by sense probe. co, column; pe, petal; se, sepal; li, lip. Bar = 50 μm