The invention discloses a dissociation method and application of rubber tree differentiated
xylem protoplast, and the dissociation method comprises the following steps: peeling off green and tender branches of a rubber tree along the stem,
cutting the stem into a plurality of sections after the
stem bark is peeled off, and immersing into enzymatic
hydrolysate; carrying out enzymolysis and light shaking at
room temperature under a dark condition for 4 hours, adding a W5 solution, carrying out light shaking, uniformly mixing, filtering, centrifuging, pouring out supernate, and retaining
protoplast precipitate; adding the
protoplast precipitate into a pre-cooled W5 solution, resuspending, washing, pouring out supernate, and retaining the protoplast precipitate; re-suspending the protoplast precipitate in an MMG solution, and placing on ice;
trypan blue dyeing is used for detecting
cell concentration and
cell activity; and loading the obtained protoplast into a W5 solution and a WI solution respectively for overnight treatment. The efficient and stable method for extracting the
xylem differentiated protoplast is established by stripping young branches of rubber trees, performing enzymolysis and differentiation on
xylem tissues to obtain the protoplast and trying different osmotic pressures.