Fig. 4

Regulatory relationships between TsVND7 and vessel-related genes in T. sinense. a Maximum likelihood tree of genes from the VNS gene family. Bootstrap percentages for the large clades are shown next to the nodes. Species name abbreviations given in Fig. S11 in additional file 1. Angiosperm sequences of different subfamilies are shown in different colors and the gymnosperm sequences are shown in black. b Confocal images of Nicotiana benthamiana leaves transiently transformed with TsVND7.2-GFP, TsVND6.1-GFP, and TsVND6.2-GFP under the control of the 35S promoter, bar = 10 μm. c Yeast one-hybrid assay showing that TsVND6.1 and TsLBD30a interact with the TsVND7.2 promotor and TsVND7.2 interacts with the TsCesA4, TsMYB, and TsXCPa promoters. d Luciferase activity assay showing that TsVND6.1 interacts with promoters of TsVND7.1 and TsVND7.2; TsLBD30a and TsLBD30b interact with the TsVND7.2 promoter; TsVND7.1 interacts with the TsCesA8 and TsXCPa promoters; and TsVND7.2 interacts with the TsMYB and TsCesA4 promoters. e Diagram showing the interaction relationship determined in Arabidopsis. f Diagram showing the interaction relationship from the results of Y1H and luciferase activity assay in c and d and supplementary Fig. S13 b–d. The orange lines show the interactions evidenced by Y1H and luciferase activity assays, the green line shows the interactions evidenced by Y1H, and blue lines show the interactions evidenced by luciferase activity assay