Plant binary expression vector for inducing embryoid body formation and plant regeneration, and construction method and application thereof

Abstract

The invention discloses a plant binary expression vector for inducing embryoid body formation and plant regeneration, which comprises BBM genes and an FLP-FRT site specific recombination system. The FLP-FRT site specific recombination system comprises FLP genes and two identical orientation FRT sites. The BBM genes and the FLP genes are positioned between the two identical orientation FRT sites, the BBM genes are controlled by a constitutive promoter, and the FLP genes are controlled by an inducible promoter. The invention further discloses a method for constructing the plant binary expression vector and a method for inducing embryoid body formation and plant regeneration by using the plant binary expression vector. The invention can not only promote embryoid body formation of a plant callus, induce formation of a transgenic plant, and remarkably improve the transformation frequency and the regeneration frequency of the plant; but also can eliminate the negative impacts of exogenous genes on the transgenic plant. The invention provides a novel genetic transformation strategy for plant species which are difficult for genetic transformation, and has wide application prospect.

Description

Technical field [0001] The invention relates to a recombinant expression vector, in particular to a plant binary expression vector that induces embryoid body formation and plant regeneration, and also relates to a construction method and application of the expression vector. Background technique [0002] The use of genetic engineering technology to introduce specific foreign genes into plant recipients to achieve the purpose of changing plant traits (such as insect resistance, disease resistance, stress resistance, etc.) and rapidly cultivating new plant varieties is an important way of modern genetic breeding. [0003] The core of plant genetic engineering is plant genetic transformation. Plant genetic transformation methods are mainly divided into two categories: that is, the use of vector system transformation (such as Agrobacterium-mediated method, liposome method, etc.) and direct genetic transformation (such as gene gun method, PEG method, electric shock method, microinject...

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Problems solved by technology

However, this method still has the following problems: (1) the transformation frequency of some important plants (such as woody plants and monocots) is low, and the transformed plants are difficult to regenerate; (2) the transformation frequency and regeneration between different varieties of the same plant The frequency difference is large; (3) Some plant varieties widely used in production are difficult to carry out genetic transformation, etc.

The study found that the main reason for the above problems is that plant regeneration is difficult, and regenerated seedlings cannot be obtained in tissue culture.

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