Agrobacterium rhizogenes-mediated robinia pseudoacacia hairy root genetic transformation method

By using Agrobacterium rhizogenes-mediated hairy root induction technology, efficient genetic transformation of Robinia pseudoacacia was achieved under non-sterile conditions, solving the problems of low efficiency and long cycle in traditional methods, and providing a new approach for gene research and molecular breeding of Robinia pseudoacacia.

CN122445720APending Publication Date: 2026-07-24BEIJING FORESTRY UNIVERSITY
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Patent Information

Application Number
CN202610949675.3
Authority / Receiving Office
CN · China
Patent Type
Applications(China)
Current Assignee / Owner
Filing Date
2026-06-29
Publication Date
2026-07-24

AI Technical Summary

Technical Problem

Existing technologies for genetic transformation of Robinia pseudoacacia have low efficiency, long cycles, and complex operations, and require high aseptic culture conditions, which seriously restricts the development of its gene research and molecular breeding.

Method used

Using Agrobacterium rhizogenes-mediated hairy root induction technology, under non-sterile conditions, the expression vector was introduced into Agrobacterium rhizogenes, and the root-stem junction of Robinia pseudoacacia seedlings was infected with the infection solution. Subsequently, the seedlings were cultured in a non-sterile growth substrate, and the culture conditions were optimized to induce hairy roots. The eGFP marker gene was used for visual screening.

Benefits of technology

It has enabled simple, efficient, and low-cost genetic transformation of Robinia pseudoacacia, shortened the cycle, and achieved a transformation rate of over 60%, providing a simple means for gene function research and molecular breeding.

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Abstract

The invention discloses an agrobacterium rhizogenes-mediated robinia pseudoacacia hairy root genetic transformation method, and belongs to the technical field of plant genetic engineering and molecular breeding. The method comprises the following steps: preparing an agrobacterium rhizogenes infection solution carrying a target gene; taking robinia pseudoacacia seedlings, and removing roots at rhizome joints of the robinia pseudoacacia seedlings to obtain explants; immersing the wound of the explant into the infection liquid for infection; and then, directly planting the infected explant in a matrix, and carrying out co-culture and rooting induction until a transgenic hairy root is formed at the wound. According to the method provided by the invention, sterile tissue culture and a complex plant regeneration process are completely abandoned, and the genetic transformation of the robinia pseudoacacia is completed under non-sterile and non-in-vitro conditions. According to the method, the conversion period can be shortened from several months to 3-4 weeks, and the positive conversion rate can stably reach 60% or above. The method is simple and convenient to operate, low in cost, efficient and stable, and can provide important technical support for functional genomics research, genetic improvement and molecular breeding of robinia pseudoacacia.
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