Preparation and instantaneous transformation method of radix bupleuri protoplast

The invention discloses a preparation and instantaneous transformation method of a radix bupleuri protoplast. The method comprises the steps of protoplast preparation and protoplast instantaneous transformation; the protoplast preparation comprises the following steps: carrying out enzymolysis treatment on radix bupleuri hypocotyl calluses by virtue of enzymatic hydrolysate under a dark condition, and purifying, so as to obtain the radix bupleuri protoplast; and the protoplast instantaneous transformation comprises the following steps: mixing the radix bupleuri protoplast with Crispr/Cas9-T3T4 plasmid, adding a polyethylene glycol transformation solution, incubating at room temperature, adding a W5 solution to terminate the reaction after incubation is finished, centrifugally collecting the radix bupleuri protoplast subjected to instant transformation, re-suspending the radix bupleuri protoplast in an MMG solution, and incubating at room temperature in a dark place. According to the method disclosed by the in