Simple and convenient agrobacterium tumefaciens-mediated sugarcane transgene method

An Agrobacterium-mediated and Agrobacterium-transformed technology, which is applied in horticultural methods, genetic engineering, plant gene improvement, etc., can solve the problems of heavy workload and complicated work procedures, and achieve strong practicability, reduce tissue culture costs, Simple operation effect

Inactive Publication Date: 2012-07-11
FUJIAN AGRI & FORESTRY UNIV
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Problems solved by technology

The above work procedures are

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Examples

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Embodiment 1

[0038] Example 1: A Simple Agrobacterium-mediated Transgenic Method of Sugarcane

[0039] A simple Agrobacterium-mediated sugarcane transgenic method, including the following steps:

[0040] 1. Preparation of bacteriostatic agent: The preparation method of bacteriostatic agent No. 1: add 20 ml of distilled water to bottle A of Eperon bactericide, dilute it with bottle B, and make up to 100 ml with distilled water, which is the antibacterial agent. Bacterial agent No. 1; preparation method of bacteriostatic agent No. 2: add 20 ml of distilled water to bottle A of Eperon bactericide, dilute it with bottle B, add 6 g of Timentin, and dilute to 100 ml with distilled water, that is For bacteriostatic agent No. 2;

[0041] 2. Disinfection of the culture container: soak the culture bottle and bottle cap in an aqueous solution with a volume ratio of bacteriostatic agent No. 1 and water of 0.3% (V / V) for 10 hours, and save for future use;

[0042] 3. Agrobacterium transformation: sel...

Embodiment 2

[0049] Example 2: A Simple Agrobacterium-mediated Transgenic Method of Sugarcane

[0050] A simple Agrobacterium-mediated sugarcane transgenic method, including the following steps:

[0051] 1. Preparation of bacteriostatic agent: with reference to the preparation of step 1 bacteriostatic agent in Example 1;

[0052] 2. Disinfection of the culture container: soak the culture bottle and bottle cap in an aqueous solution with a volume ratio of bacteriostatic agent No. 1 and water of 0.5% (V / V) for 10 hours, and save for later use;

[0053] 3. Agrobacterium transformation: with reference to step 3 Agrobacterium transformation in Example 1;

[0054] 4. Medium preparation: co-cultivation medium is 1 / 2MS+3.0 mg / L 2,4-D+100 μmol / L acetosyringone+5 ml / L bacteriostatic agent No. 2+20.0 g / L sucrose+3.5 g / L agar powder, pH5.8; selection medium is MS+3.0 mg / L 2,4-D+3 ml / L bacteriostatic agent No. 2+30.0 g / L sucrose+3.5g / L agar powder, pH5 .8; the screening medium is MS+2 mg / L 6-BA+0.2 ...

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Abstract

The invention relates to a simple and convenient agrobacterium tumefaciens-mediated sugarcane transgene method. The transgene method comprises the steps of preparing a bacteriostatic agent, disinfecting a culture container, transforming agrobacterium tumefaciens, preparing a culture medium, co-culturing, selectively culturing, culturing by screening, culturing strong seedlings and culturing by rooting, wherein the bacteriostatic agent can be simply prepared; the culture medium can be prepared by preparing the required components according to different conventional methods of culture medium formulae and adding the bacteriostatic agent; both the culture container and the culture medium do not need to be sterilized by high temperature and high pressure, so that in a process of preparing the culture medium of the agrobacterium tumefaciens-mediated sugarcane transgene, the working amount and the energy consumption are greatly reduced, the tissue culture process is simplified, and the tissue culture cost is lowered; compared with the conventional agrobacterium tumefaciens-mediated sugarcane transgene method, the cost can be reduced by above 10%. The simple and convenient agrobacterium tumefaciens-mediated sugarcane transgene method is simple to operate, strong in practicality and good in popularity.

Description

technical field [0001] The invention relates to a plant transgenic method, in particular to a simple Agrobacterium-mediated sugarcane transgenic method, which belongs to the field of biotechnology. Background technique [0002] Genetic engineering refers to the creation of target organisms that are more in line with people's needs in a relatively short period of time through rigorous experimental design, using in vitro DNA recombination and transgenic technologies, according to people's wishes. With the development of modern molecular biology theory and the advancement of research technology, it is possible to use genetic engineering technology to carry out directional improvement of biological varieties and breeding of target traits. Among them, the use of transgenic technology to improve crop varieties has unique advantages. Among them, there are mainly two kinds of transgenic technology in the narrow sense, one is the gene gun-mediated method, and the other is the Agrobac...

Claims

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Application Information

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IPC IPC(8): C12N15/84A01H4/00A01H5/00
Inventor 阙友雄林庆良许莉萍高世武陈如凯
Owner FUJIAN AGRI & FORESTRY UNIV
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