Fig. 2

Cloning of PHS39. A ∆(SNP-index) plot of all mutated sites on chromosome 6 by MutMap + analysis. The red, green, and purple curves represent the average ∆(SNP-index), the 95%, and 99% thresholds, respectively. B Five mutated SNPs with ∆(SNP-index) ≥ 0.6 were located in five candidate genes. C Schematic representation of the candidate gene LOC_Os06g44920. This gene contains only one exon. The mutated nucleotide is highlighted in red. D The protein structure of LOC_Os06g44920. The mutated amino acid is highlighted in red. The F-box and DUF 295 are represented as the functional domains in green and blue, respectively. E Schematic representation of the candidate gene LOC_Os06g45480. Gray boxes, white boxes, and black lines represent exons, UTRs, and introns, respectively. The underlined GU-AG sequence identified at RNA splicing was abolished by the G-to-A substitution in the phs39 mutant. The dotted line to the left of TAA represents the omitted 45 bp bases, and the dotted line to the right of TAA represents the omitted 53 bp bases. The mutated nucleotide is highlighted in red. F The protein structure of LOC_Os06g45480. The protein contains a transmembrane region (green box) and a peptide_M20 domain (blue box). The red box represents 15 amino acids that differ between the WT and phs39 mutant. ‘*’ indicates the termination of protein translation. G The expression level of OsAAH in various tissues was detected by RT‒qPCR. Values represent the means ± SD (n = 3). H GUS staining in seeds at the filling stage (I-III), seed germinated after 2 d (IV), 10-day-old seedling (V), and leaf (VI), spikelet (VII), root (VIII), and anther (IX) at the heading stage driven by the OsAAH promoter. All bars are 1 mm except for 5 mm in (V). I Subcellular localization of the OsAAH-GFP fusion protein in tobacco leaf epidermal cells using mCherry-HDEL as an endoplasmic reticulum localization marker. The GFP was used as the control. Scale bars = 20 μm