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Method for cultivating transgenic sycamore plant mediated by agrobacterium

An Agrobacterium-mediated, transgenic technology, which is applied in the field of garden tree genetic engineering, can solve the problems of low expression level of exogenous genes, multiple transformations, and low frequency of gene transformation

Inactive Publication Date: 2007-01-24
HUAZHONG AGRI UNIV
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  • Application Information

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Problems solved by technology

Nevertheless, compared with crops, gene transformation technology of woody plants still has many problems, such as low frequency of gene transformation, difficulty in regeneration of transgenic plants, few target genes available from woody plants themselves, currently obtained In transgenic plants, there are many transformations from sexual materials, and the expression level of exogenous genes in transformed plants is low or unstable. further in-depth research
[0005] So far, there have been no reports on the cultivation of transgenic plants of Platanus at home and abroad

Method used

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  • Method for cultivating transgenic sycamore plant mediated by agrobacterium
  • Method for cultivating transgenic sycamore plant mediated by agrobacterium
  • Method for cultivating transgenic sycamore plant mediated by agrobacterium

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Experimental program
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Effect test

Embodiment 1

[0058] Embodiment 1: the breeding method of Platanus transgenic plant

[0059] 1. Transformation receptor material and culture method

[0060] The syringa seeds were collected from the campus of Huazhong Agricultural University in Wuhan City, Hubei Province, China, and the in vitro seedlings induced by hypocotyls were used as experimental materials. For the culture methods of tube plantlet proliferation, rooting and leaf regeneration, refer to Liu Guofeng and Bao Manzhu (Liu G, Bao M, Adventitious shoot regeneration from in vitro cultured leaves of London plane tree (Platanus acerifolia Willd.) Plant Cell Rep, 2003, 21: 640-44 ) method of reporting.

[0061] Unless otherwise specified, the medium components of the present invention are shown in Table 1. Adjust the pH value to 5.8-6.0, and sterilize according to conventional methods.

[0062] 2. Transformation carrier material and culture method

[0063] Contains the reporter plasmid pCAMBIA2301 (see Genebank, gene accessio...

Embodiment 2

[0107] Embodiment 2: comparative test embodiment

[0108] 1. Experiment of effective selection pressure of Kanamycin (Kan)

[0109] Using the aseptic leaves of Platania suzuki as explants, the selection pressure of kanamycin (Kan) was determined for its regeneration. Inoculate the scratched Platanus leaves on the differentiation medium C (see Table 1) with different concentrations of Kan, and compare the inhibitory effects of different concentrations of antibiotics on the differentiation of adventitious buds induced by explants, so as to determine the appropriate screening concentration for genetic transformation . According to the results of the preliminary experiment, five kinds of Kan concentration gradients of 0, 12.5, 20, 25, and 50 mg / L were set up for the explants of Platania suzuki leaves. A new medium was replaced every 15 days, and the browning rate and differentiation rate of the explants were counted after 60 days.

[0110]In the process of plant genetic transfo...

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Abstract

The present invention belongs to the field of plant transgene technology, and is especially agrobacterium mediating method of cultivating transgenic plant of sycamore as one garden tree. The agrobacterium mediating method includes the steps of genetic transformation, PCR detection and Southern hybridization detection, and features that through agrobacterium mediation to transduce GUS reporter gene to acceptor sycamore, adding kanamycin as selecting agent to the basic culture medium, screening the transformed acceptor cell, PCR detection, GUS gene detection, Southern hybridization and other steps, transgenic plant of sycamore is obtained. The present invention provides technological platform for the genetic improvement of tree, especially sycamore.

Description

technical field [0001] The invention belongs to the technical field of genetic engineering of garden trees. In particular, it relates to a method for cultivating transgenic plants of Platanus chinensis mediated by Agrobacterium. Background technique [0002] Woody plants are mostly perennial plants. Compared with crops and other annual and biennial herbaceous plants, they generally have the characteristics of long life cycle, late maturity and flowering, and complex genetic background (highly heterozygous genes). Therefore, it is difficult to achieve the expected goal in a short period of time by using conventional breeding methods to improve it, resulting in slow progress in woody plant genetics and breeding. In recent years, the development of plant genetic engineering and its application in woody plants has brought infinite vitality to the genetic improvement of woody plants. [0003] In general, due to the limitations of the charact...

Claims

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Application Information

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IPC IPC(8): C12N15/82
Inventor 包满珠刘国锋李志能方芳张俊卫
Owner HUAZHONG AGRI UNIV
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