Generation of rice mutants using the CRISPR/Cas9 method
For the generation of perox4 and osmt1a/b knockout lines, guide RNAs (gRNAs) (Supplemental Table 4) were designed to target exons of the targeted genes (Fig. S3). The synthesized oligo of gRNAs was annealed to form the oligo adaptors, which then was joined to the BGK03 CRISPR/Cas9 vector (Biogle Biotechnology, Hangzhou, China). The CRISPR/Cas9 plasmids were introduced into Agrobacterium tumefaciens EHA105 and then into rice throughAgrobacterium- mediated transformation as described previously. The individual T1 plants were genotyped by sequencing the DNA products of PCR with primers (Supplemental table 4).