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Method for transforming stem tips of plants and special tool thereof

A plant and shoot tip technology, which is applied in the field of plant shoot tip transformation methods and special tools, can solve the problems of difficult batch genetic transformation, low callus induction rate and plant regeneration rate, and achieves overcoming genotype dependence and body regeneration. The effect of cell mutation, improving the transformation efficiency of shoot apex, and simple operation

Inactive Publication Date: 2012-01-18
INST OF BOTANY CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the callus induction rate and plant regeneration rate of this system are very low, and it is difficult to be used for batch genetic transformation.

Method used

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  • Method for transforming stem tips of plants and special tool thereof
  • Method for transforming stem tips of plants and special tool thereof
  • Method for transforming stem tips of plants and special tool thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0058] Embodiment 1, utilize wheat stem tip transformation system to obtain transgenic wheat

[0059] 1. Establishment of wheat shoot tip transformation system

[0060] 1. Materials

[0061] A. Wheat varieties:

[0062] Middle wheat 12 wheat (hereinafter referred to as wild-type wheat), winter in nature; strong tillering ability, high spike rate; spindle-shaped spikes, long awns, white hulls, white grains, and horny kernels; the number of spikes per mu and spikes in yield factors Large number of grains, low thousand-grain weight; short plant, about 65 cm high in water-saving area, about 70 cm in high-fertilization area, good lodging resistance; good cold resistance; resistant to, slow stripe rust, slow leaf rust Rust, powdery mildew and stem rust.

[0063] Purchased from the Seed Sales Department of Beijing Zhongnong Gold Land Agricultural Development Research Center.

[0064] B. Transfection of recombinant bacteria: C58 / pCAMBIA3300-ubinos

[0065] Recombinant Agrobacteri...

Embodiment 2

[0161] Embodiment 2, Utilize Salicornia shoot tip transformation system to obtain transgenic Salicornia

[0162] 1. Experimental group

[0163] 1. Materials

[0164] A. Salicornia (S. europeae L.) seeds (hereinafter referred to as wild-type Salicornia) were purchased from Jinglong Ocean Industry Development Co., Ltd., Dafeng City, Jiangsu Province.

[0165] B. Transfection of recombinant bacteria: C58 / pCAMBIA3300-ubinos

[0166] Recombinant Agrobacterium C58 / pCAMBIA3300-ubinos is a recombinant bacterium obtained by transferring the named plasmid pCAMBIA3300-ubinos into Agrobacterium C58.

[0167] Plasmid pCAMBIA3300-ubinos contains ubiquitin promoter and selection marker gene encoding phosphinothricinacetyltransferase (PAT, phosphinothricinacetyltransferase) gene bar.

[0168] Agrobacterium strain C58 was disclosed in the document "Overexpression of Organellar and Cytosolic AtHSP90 in Arabidopsis thaliana Impairs Plant Tolerance to Oxidative Stress. Hongmiao Song, Pengxiang...

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Abstract

The invention discloses a method for transforming stem tips of plants and a special tool thereof. The method for cultivating a transgenic plant comprises the following steps of: 1) puncturing a growing point of a stem tip of a target plant by using a metal brush to obtain a treated target plant; 2) impregnating the treated target plant obtained in step 1) by using a bacterial liquid containing target deoxyribonucleic acid (DNA) to obtain an impregnated seedling; and 3) sequentially coculturing and continuously culturing the impregnated seedling obtained in 2) to obtain the transgenic plant containing the target DNA. Through the experiment, a set of efficient and quick plant transformation system is established by taking meristems of stem tips of seedlings of a monocotyledon, namely wheat and a dicotyledon, namely Salicornia europaea as transformation receptors of Agrobacterium tumefaciens respectively and adopting measures such as retrofit of stem tip damage tools, research on the seedling ages of the seedlings, vacuum-assisted infiltration treatment, application of infiltration promoters and the like, so that the transformation efficiency of the stem tips is improved.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a plant shoot tip transformation method and a special tool thereof. Background technique [0002] The plant genetic transformation method is to introduce foreign genes into recipient cells in a specific way to cause directional and permanent genetic variation. At present, people have established a variety of transformation systems. For most genetic transformation systems, the genetic transformation method is the key link in the genetic transformation system, which determines the success or failure and efficiency of the transformation. There are many transformation methods for plants: Agrobacterium-mediated method, particle gun method, electric shock method, PEG method, etc. Agrobacterium-mediated method and particle gun method have become the main methods for genetic transformation. [0003] In the past, it was thought that Agrobacterium-mediated genetic transformation was limited...

Claims

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Application Information

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IPC IPC(8): C12N15/82C12N1/21A01H5/00
Inventor 李银心吕素莲
Owner INST OF BOTANY CHINESE ACAD OF SCI
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