Agrobacterium-mediated genetic transformation method for sea barleygrass

An Agrobacterium-mediated genetic transformation method for sea barleygrass, comprising:

• step S1, selection and sterilization of immature embryo materials of sea barleygrass, which comprises:
• selecting the immature embryo materials of sea barleygrass with immature embryos each having a length in a range of 0.5-1.0 millimeters (mm), and sterilizing selected immature embryo materials with alcohol and sodium hypochlorite to obtain sterilized seeds;
• step S2, separation and callus induction of the immature embryos, which comprises:
• separating the immature embryos from the sterilized seeds, crosscutting the immature embryos, and inducing callus generation and proliferation, thereby obtaining calli;
• step S3, Agrobacterium infection and selective culture, which comprises:
• adjusting pre-culture time and Agrobacterium infection time of the calli based on the callus generation and Agrobacterium growth to thereby prevent excessive Agrobacterium liquid;
• wherein a culture medium for the selective culture is added with a mixture of ticarcillin and clavulanic acid and hygromycin;
• and
• step S4, callus differentiation, plantlet formation, and positive plantlet detection, which comprises:
• performing adventitious bud induction culture and rooting induction culture under a shielding-formed low-light environment to thereby obtain tissue culture plantlets, and then performing deoxyribonucleic acid (DNA) extraction and polymerase chain reaction (PCR) detection on obtained tissue culture plantlets to obtain PCR detection results, and determining whether target fragments are amplified based on the PCR detection results, to thereby determine whether the tissue culture plantlets are positive plantlets;
• wherein an Agrobacterium tumefaciens recombinant strain used for transformation carries a gene editing vector, further comprising performing PCR amplification on a sequence near a target of each of the positive plantlets, and detecting mutation sites by Sanger sequencing, and a hmsos1sg1F sequencing primer consisting of the nucleotide sequence comprising SEQ ID NO:
• 4.