The invention discloses a method for prolonging the subculturing interval time of
tissue culture seedlings of apple. A culture medium comprises the following components: 0.5-0.75 mg / L of MS+BA, 0.05 mg / L of NAA, 40.0 g / L of white granulated
sugar and 6.0 g / L of
agar; the packing quantity is 40-50% more than that in conventional breeding, and a
bottleneck is sealed by using a plastic sealing film;
tissue culture seedlings of apple are transferred to the culture medium and slowly grow about 2 weeks under the culture conditions that the temperature is 22-28 DEG C, the illumination intensity is 2000 lx, and the illumination time is 10-14 h / d, and then the
tissue culture seedlings of apple are placed in a
chemical agent storage box with a temperature of 2-8 DEG C so that the growth is slowed down; and when the culture medium is approximately dried, the tissue culture seedlings of apple are taken out of the refrigerated container to grow normally for 1-2 weeks or are directly transferred to a new culture medium. In such a way, the subculturing interval time of tissue culture seedlings of apple can be successfully prolonged from 2-3 months to 15-18 months, the
survival rate of stem tips is more than 90%, the
workload of subinoculation is greatly reduced, and the
mutation probability is also reduced. The technique is applied to the
germplasm preservation of plantlets of apples and other parts of orchardfruits and flowers, is simple, convenient, saving, and high in efficiency, and provides a new way for the preservation of
germplasm resources.