Method for prolonging subculturing interval time of tissue culture seedlings of apple

Abstract

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.

Description

technical field [0001] The invention relates to a method for in vitro preservation of apple germplasm resources using tissue culture technology. Background technique [0002] With the rapid growth of population, unreasonable reclamation of wasteland, large-scale deforestation, and industrial pollution, the environmental ecological balance has been seriously damaged, and some precious and rare species are facing extinction or are on the verge of extinction. At the same time, due to the development of plant breeding technology, artificial selection and the promotion of improved varieties, the phenomenon of single species composition is becoming more and more obvious, resulting in the loss of many beneficial and precious germplasm resources. Therefore, the conservation of plant germplasm resources has become a global concern. [0003] There are two main methods for the preservation of plant germplasm resources. One is in situ preservation, that is, establishing nature reserve...

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Problems solved by technology

[0008] The technical scheme adopted by the present invention to solve its technical problems is: the medium of apple tissue culture seedlings is slightly adjusted to delay growth, and after inoculation, through a period of slow growth of seedlings, the culture conditions of 25 ± 3 ° C are changed to 8 ± 1 °C. The low temperature conditions around ℃ can effectively inhibit the growth of test tube seedlings; through experiments, we can screen out suitable sealing materials for tissue cultured seedlings, solve the problem of high humidity in the refrigerator and easy to grow mold pollution, and prolong the storage time of apple tissue cultured seedlings

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