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Easy and efficient agrobacterium tumefaciens induced corn gene transfer method

An Agrobacterium-mediated, transgenic technology, applied in plant genetic improvement, botanical equipment and methods, applications, etc., can solve the problems of tissue culture, callus differentiation difficulties, application limitations, and low transformation efficiency, and achieve unique application value. , Easy to operate, avoid the effect of complicated process

Inactive Publication Date: 2005-04-06
ANHUI AGRICULTURAL UNIVERSITY
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AI Technical Summary

Problems solved by technology

At the same time, the tissue culture and callus differentiation of maize are more difficult than those of rice, wheat and other plants. Maize tissue culture is more restricted by genotype and the range of receptors is smaller. The application of gun method, ultrasonic wave and electric shock method on maize has been greatly limited, and although the pollen tube channel method can avoid the tissue culture process and directly obtain transformed seeds, the transformation efficiency is low. Therefore, it is not necessary to establish and develop a new method. Simple and efficient maize transgenic technology through tissue culture is imminent

Method used

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Experimental program
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Embodiment Construction

[0010] The technological process of the present embodiment method is as follows:

[0011] Exogenous geneTi plasmid expression vector construction→Agrobacterium transformation solution preparation→in situ transformation→transformant screening→transformant identification→transgenic plants.

[0012] The specific implementation steps are as follows:

[0013] 1. Determination of the target gene and vector construction.

[0014] The relevant target gene is selected, and the maize expression vector is constructed using Ti plasmids such as pCAMBIA1300 or pBI121.

[0015] 2. Preparation of Agrobacterium transformation solution.

[0016] The expression vector is introduced into the competent cells of Agrobacterium by a three-parent hybridization method or a direct transformation method to obtain transformed Agrobacterium strains.

[0017] (1) Three-parent hybridization method, each of the three cultured bacterial solutions of Agrobacterium tumefaciens LBA4404, Escherichia coli HB101...

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Abstract

Way to change the genetic of corn with farming bacilli. The character is using the farming bacilli with other genetic to infect the female young fringe and flower of corn take transform of the corn. The transform of the corn is damnificated the flower and female young fringe of the corn first, then put the farming bacilli with objective genetic lay on or dripping on it, repeat it 3 to 4 times, through the filter of fastness and examine of molecule to the seminal, obtain the plant with changed gene. This invention avoid the tissue culture and reformat process of the plant, is simple and short cut, the inverted offspring can individual plant directly.

Description

Technical field: [0001] The invention relates to a method for transgenic maize. Background technique: [0002] Applying transgenic technology to introduce the target gene into a suitable recipient to obtain stable transgenic plants is an important part of transgenic. In the study of corn transgenics, a variety of transgenic technologies have been pioneered and established, including gene gun method, PEG method, electric shock method, ultrasonic-mediated method, pollen tube channel method, etc., which are respectively adapted to different receptors and have achieved certain results. However, these techniques have various defects in different aspects such as tissue culture, receptor selection, and transformation efficiency, which bring a lot of inconvenience to the experimental operation. [0003] As we all know, the workload of tissue culture is relatively large, requiring a large number of professionals to do heavy work. The traditional Agrobacterium method is based on tis...

Claims

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Application Information

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IPC IPC(8): A01H1/00
Inventor 程备久朱苏文项艳张永风王洲谈应权马庆韩国民叶辉程郢
Owner ANHUI AGRICULTURAL UNIVERSITY
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