Additional file 1: of The transcription factor OsbHLH035 mediates seed germination and enables seedling recovery from salt stress through ABA-dependent and ABA-independent pathways, respectively

Figure S1. Abiotic stress-responsive OsbHLHs. The fold change among each OsbHLH gene under different abiotic stresses is calculated as a ratio normalizing the data to it corresponding nontreated control. Red and green colors represent up- and downregulation, respectively. C, cold stress; D, drought stress; S, salt stress; and H, heat stress. Figure S2. Prediction of the conserved domains in OsbHLH035 and characterization of the NG7221 mutant line. (A) The ScanProsite tool in ExPASy ( http://www.expasy.org /) predicts the presence of a typical bHLH domain (residues 64 to 113) in OsbHLH035. (B) Based on its annotation in the rice Tos17 insertion mutant database ( https://tos.nias.affrc.go.jp /), NG7221 is a single retrotransposon insertional line. (C) The identification of homozygous NG7221 mutants via PCR genotyping of gDNA. The primer positions and sequences are shown in Fig. 1a and Additional file 2: Table S1, respectively. Arabic numbers in (C) represent three independent biological replicates within each genotype. Figure S3. GFP-fused OsbHLH035 protein is predominantly localized to the nucleus in rice calli. The husk-removed seeds harboring OsbHLH035::GFP-OsbHLH035 were placed on callus induction medium (Tran and Sanan-Mishra 2015) for 7 days and then the calli were subjected to GFP visualization by a confocal microscopy. Scale bars, 10 μm. Figure S4. The endogenous ABA contents in both germinating WT and Osbhlh035 seeds. The husk-removed seeds were grown on basal medium for 2 days and then subjected to ABA ELISAs. Figure S5. Phylogenetic analysis of AtCYP707As and OsABA8oxs using the neighbor-joining method. Numbers next to the descendant indicate confidence values based on the bootstrap method. (PDF 570 kb