Supporting information
Table S1. Primers used for qRT-PCR.
Table S2. Primers used for vector construction and positive hairy roots line selection.
Figure S1. Phylogenetic tree of SmGRAS5 transcription factor. The phylogenetic tree was constructed by the maximum-likelihood method of MEGA 6.0. Members in the same sub-branch were marked by a circle filled with the same color.
Figure S2. Transactivation activity of SmGRAS5. Yeast AH109 containingpGBKT7-53+pGADT7-T (positive control), pGBKT7 (negative control) and SmGRAS5-pGBKT7 were spotted onto SD/-Trp and SD/-Trp-His-Ade medium, respectively.
Figure S3. Verification of positive hairy roots lines by using PCR. PCR screening of SmGRAS5 overexpressing or antisense expressing lines.
Figure S4. Growth phenotypes of SmGRAS5 transgenic hairy roots lines.
Figure S5. GA induces SmGRAS5 responses.
(A) Relative quantitative analyzes the expression of SmGRAS5 with 100 μM GA treatment for 2/24 h. (B) Promoter analysis of the GA response element of SmGRAS5.
Figure S6. Y1H assay of SmGRAS5 and SmCPS1 promoter.SmCPS1 promoter+pGADT7 as the negative control andSmCPS1 promoter+pGADT7-SmERF6 as the positive control.
Figure S7. Purified MBP protein and SmGRAS5 protein with MBP label were verified via Western blot. M: protein marker.
Figure S8. EMSA analysis of the interaction of SmGRAS5 with the promoter of GA biosynthesis genes.
(A) EMSA analysis of the interaction of SmGRAS5 with CPS5 promoter. (B) EMSA analysis of the interaction of SmGRAS5 with KS promoter. (C) EMSA analysis of the interaction of SmGRAS5 with GA3ox1 promoter.
Figure S9. Validating the different expression levels of the identified genes from RNA-seq data by qRT-PCR. Expression of the representative GA-responsive genes in the RNA-seq experiments of G5O14. The expression of genes in the untreated ATCC hairy roots lines was set to 1. Standard errors were calculated from three sets of biological replicates. Significant differences using one-way ANOVA and S-N-K comparison tested,P < 0.05.