Method for improving regeneration efficiency of apple transgenic material

Abstract

The invention discloses a method for improving the regeneration efficiency of an apple transgenic material. The method comprises the steps of culturing a tissue culture seedling of bredia rotundifolia in an expanding propagation culture medium for a month, cutting off callus at a stem base part by using a scalpel, then transplanting a regenerated stem section into a new culture medium; culturing the regenerated stem section in the expanding propagation culture medium for a month, transversely cutting the midrib of a fully-stretched functional leaf for two or three times, infecting by using agrobacterium, and then inoculating into a predifferentiation culture medium; carrying out dark culture for 2-4 days, and then transferring to a differentiation culture medium; after the dark culture is carried out for two weeks, culturing under light until resistant buds are differentiated; transplanting the resistant buds into a subculture medium, and further culturing for 20-30 days; selecting regenerated plantlets with the height of 1.5cm, and transplanting the selected regenerated plantlets into a rooting medium, culturing for a month, and transplanting. After the method is adopted, the detached leaf regeneration efficiency reaches 80.33%, the expanding propagation coefficient reaches 8.2, and the rooting rate reaches 96.38%.

Description

technical field [0001] The invention belongs to the technical field of apple planting, and in particular relates to a method for improving the regeneration efficiency of apple transgenic materials. Background technique [0002] In recent years, the development of genetic engineering has provided new technical methods for the genetic improvement of apple varieties, broadened the source of genes, broken through the boundaries between species, and can isolate various functional genes from animals, plants and microorganisms for genetic improvement, shortening the time of breeding. cycle, improve breeding efficiency, and open up a new technical approach for the research of apple genetics and breeding and the utilization of various genetic germplasm resources. Foreign countries successfully applied the icp gene to transform apples as early as 1991. In 1992, the United States approved the field test of transgenic apple plants carrying the icp gene. Domestically, Daketon also used t...

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Problems solved by technology

[0004] The purpose of the present invention is to provide a method for improving the regeneration efficiency of apple transgenic materials, aiming to solve the problem that the existing apple transgenic materials are Royal Gala and M26, the transgenic materials are single, and the efficiency of leaf regeneration plants is low

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