4.2. CRISPR/Cas9 Vector Construction and Plant Transformation

The CRISPR/Cas9 binary vector pHUE411 carrying two gRNAs targeting (5′-AGGACTCACGGTTGCAGCGG-3′; 5′-AGGCAAGGCCTGTAGTTCCT-3′) was transformed into Agrobacterium strain EHA105, and Agrobacterium-mediated method was used to transform immature embryos of Zhenggeng 1925. The genomic DNA was extracted from transgenic seedlings, and the primers for cloning fragments with targets were listed in Table S3. At last, the PCR products were sequenced and blasted to identify homozygous mutants.