Figure 3
Sequence analysis and phenotypic observation of OsSTRL2 CRISPR/Cas9-mediated mutant. (A) Gene structure of OsSTRL2 and mutation analysis of OsSTRL2 gene in transgenic plants. The sequence (5ʹ-CCTGCTCCTCACCGACCCGTTCC-3ʹ) located in the first exon of the OsSTRL2 gene was selected as the target site of sgRNA. The black boxes indicate the exons. The red characters indicate the PAM. The green characters indicate the four different types of mutation events generated by CRISPR-Cas9 in the mutants (The detailed information of four different mutation types are listed in Supplementary table S2). (B) Putative amino acid sequences alignment of four types of mutation as described in Fig. 3A. The sequences were aligned using DNA-MAN and displayed using BOXSHADE. The deep blue boxes indicate the 100% identities of amino acids. (C) Phenotypic comparison of the WT (left) and the mutant plant (right) at the maturity stage. (D) Panicle comparison of the WT (left) and the mutant (right) at the heading stage. (E) Floral comparison of the WT (left) and the mutant (right) (with the palea and lemma removed) at the heading stage. (F and G) I2–KI pollen staining for the WT and the mutant. Bars = 0.20 mm (D and E); 30 μm (F and G).
