METHOD FOR KNOCKING OUT BNMAX1 GENE IN BRASSICA NAPUS L. USING CRISPR-CAS9 SYSTEM AND APPLICATION

摘要

Provided are a method for knocking out BnMAX1 gene in Brassica napus L. using CRISPR-Cas9 system and an application, two sgRNAs of specifically targeting Brassica napus L. BnMAX1 gene are designed and synthesized into oligo dimer, which are connected with Cas9 carrier and introduced into hypocotyl callus of Brassica napus L. by Agrobacterium-mediated genetic transformation technology to regenerate into shoots, Cas9 nuclease cleaves target sequence under the guidance of sgRNA, each sgRNA can mediate the cleavage of the BnMAX1 gene on the A and C genomes through the CRISPR-cas9 system, achieving the purpose of knocking out gene. Phenotypic identification shows that homozygous mutant lines increase the number of branches and pod number per plant, decrease plant height and increase yield.