Agrobacterium rhizogenes mediated spraying decoration transgenosis method and application

An Agrobacterium-mediated, Agrobacterium-based technology, applied in the field of transgenics, can solve problems that have not been reported, and meet the requirements of extensive conditions, large practical value, and short transformation cycle

Inactive Publication Date: 2011-03-09
THE INST OF BIOTECHNOLOGY OF THE CHINESE ACAD OF AGRI SCI +1
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  • Claims
  • Application Information

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Problems solved by technology

The method is simple to operate, can be operated directly under field environmental conditions, and possesses all the advantages of the genetic transformation method of Agrobacterium-mediated transgenic seedlings obtained through the regener

Method used

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  • Agrobacterium rhizogenes mediated spraying decoration transgenosis method and application
  • Agrobacterium rhizogenes mediated spraying decoration transgenosis method and application
  • Agrobacterium rhizogenes mediated spraying decoration transgenosis method and application

Examples

Experimental program
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Embodiment

[0025] Example: Transformation of Agrobacterium-mediated Spraying to Obtain Cotton Transgenic Plants

[0026] 1. Materials and methods

[0027] 1) Cotton material: The cotton material is selected from the optional variety Y18R.

[0028] 2) Strain: Agrobacterium LBA4404.

[0029] 3) Vector: pGBIF4ABC-EPSPS ( figure 1 ). The plant expression vector constructed in our laboratory contains two independent expression cassettes, both of which are promoted by 35S Promoter and the terminator is Nos. The gene encoded by the first expression cassette is the glyphosate resistance gene EPSPS, and the gene encoded by the second expression cassette is the fusion gene of the Bt gene and the Cpti gene.

[0030] 4) Tool enzymes and modification enzymes: various restriction enzymes and modification enzymes were purchased from TaKaRa Company, NEB Company and Fermentas Company.

[0031] 5) Chemical reagents: All chemicals are analytically pure at home and abroad.

[0032] 6) Primer synthesis...

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Abstract

The invention relates to an agrobacterium rhizogenes mediated spraying decoration transgenosis method. In the method, a process that the agrobacterium rhizogenes suspension is directly sprayed on the stamen before pollen-shedding stage and the stamen in the pollen-shedding stage under the condition of living body in the field environment is adopted, thus having simpler operation and avoiding the damage to the organ of flower at the same time. Calculated as the number of the transgenosis plant/transforming flower, the transformation efficiency in the cotton can be 10%. The method of the invention has great practical application value in the plant genetic transformation and molecular breeding. The transforming material used in the invention can be Malvaceae cotton, or monocotyledonous and dicotyledoneae seedling plants such as rice, corn, wheat, soybean, rapeseed plant, cucumber, tomato, tobacco, fruit trees, forest, flowers and officinal plants and the like, and has a certain application value in the genetic transformation and molecular breeding of the monocotyledonous and dicotyledoneae seedling plants.

Description

technical field [0001] The invention relates to cotton genetic engineering, in particular to a transgenic method for transforming cotton through the method of spraying flowers mediated by Agrobacterium. Background technique [0002] Plant genetic engineering is the application of recombinant DNA technology to introduce foreign genes into recipient plant cells, so that recipient plants can obtain new genetic traits. In the 1980s, on the basis of a series of major advances in the fields of cell and tissue culture and molecular biology research, plant genetic engineering has developed rapidly. Cotton is an important economic crop in my country and plays a decisive role in the development of agriculture and cotton spinning industry. However, traditional breeding methods are mostly limited to inter-species or inter-species chromosome recombination, which has great limitations. Therefore, it is of great significance to use genetic engineering technology to breed new cotton variet...

Claims

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Application Information

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IPC IPC(8): C12N15/82A01H5/00
Inventor 郭三堆孟志刚张锐石雅丽
Owner THE INST OF BIOTECHNOLOGY OF THE CHINESE ACAD OF AGRI SCI
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