The invention discloses a preparation and
conversion method of soybean protoplasts, and relates to the technical field of
plant gene engineering. The method comprises the following steps of (1) performing induction to obtain soybean embryogenic calluses; (2) adding an
enzyme solution, performing enzymolysis on the embryogenic calluses, performing
filtration and centrifuging, adding a W5-23% solution, and performing centrifuging so as to obtain purified protoplasts; and (3) putting the purified protoplasts on ice, performing treatment, performing re-suspending with MMG suspension liquid, then sequentially adding plasmids and a PEG4000 solution, performing uniform mixing, performing standing, adding a W5 solution, ending a
conversion reaction, performing re-suspending on protoplasts obtained through centrifuging with a WI solution, performing shifting into a culture dish, and performing light-away room-temperature putting. According to the preparation and
conversion method disclosed by the invention, the compounding ratio of the
enzyme solution and PEG instantaneous
conversion reaction conditions are optimized, the yield and the conversion rate of the soybean protoplasts are significantly increased, identification of soybean
gene functions is facilitated, and a reliable basis is further provided for exploration of the preparation and conversion conditions of the protoplasts of other plants and other positions.