Agrobacterium rhizogenes transformation method for living body growth cone of woody plant

Abstract

The invention discloses an agrobacterium rhizogenes transformation method for a living body growth cone of a woody plant, which comprises multiple steps of preparing agrobacterium rhizogenes engineering bacterium osmosis transformation liquid, converting and inoculating the living body growth cone, coculturing the growth cone and agrobacterium rhizogenes engineering bacteria, screening resistant sprouts, culturing resistant branches, identifying a transgenic plant, and the like. The invention breaks through two technical bottlenecks of difficult gene transformation and difficult regeneration of the woody plant, is independent of plant genotype and tissue culture, has the advantages of high transformation efficiency, simple operation, low cost, and the like and is suitable for most of woody plants, such as tea trees, and the like, especially the woody plant with full terminal buds.

Description

technical field [0001] The invention relates to a plant gene transformation method, in particular to a method for transforming agrobacteria in living bodies of woody plants such as tea trees. Background technique [0002] Plant genetic transformation refers to the introduction of exogenous genes into the recipient plant genome through a certain technology or approach, so that it can be stably inherited in the recipient plant, and endow the plant with new agronomic traits, such as high resistance, high yield, high quality, metabolic Significant changes in substance content, etc. [0003] In plant genetic transformation, there are usually two types of gene receptors: in vitro culture materials and living materials. Using in vitro culture materials such as protoplasts, suspension culture cells, callus, microspores and original explants as gene recipients, that is, to achieve genetic transformation through tissue culture and plant regeneration, is currently the most important m...

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Problems solved by technology

In addition, woody plants generally contain a large amount of polyphenols. Polyphenols accumulate rapidly and massively in wound cells, which is a self-protection mechanism for plants. Gene transformation has an inhibitory effect, and even has a killing effect on Agrobacterium itself, which leads to problems such as low transformation frequency, unstable effect, and difficulty in reproducibility of the Agrobacterium transformation method in the genetic transformation of woody plants based on tissue culture and plant regeneration.

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