Soybean genetic transformation method

A genetic transformation method and genetic transformation technology are applied in the field of genetic transformation and high-efficiency regeneration of soybean cotyledon nodes, which can solve the problems of large explants, long regeneration time, and large differences in regeneration frequency, and achieve simple cultivation process and short regeneration time. short effect

Inactive Publication Date: 2011-07-20
INNER MONGOLIA AGRICULTURAL UNIVERSITY
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  • Abstract
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  • Application Information

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Problems solved by technology

Although cotyledonary nodes have many advantages as explants, there are also disadvantages such as large explants, large differences in regeneration frequency of different soybean genotypes, long regeneration time, and difficult screening.

Method used

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  • Soybean genetic transformation method
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  • Soybean genetic transformation method

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Experimental program
Comparison scheme
Effect test

Embodiment Construction

[0015] The experimental procedures of the present invention will be described in detail below in conjunction with specific embodiments:

[0016] 1. Experimental materials

[0017] 1. Plant material: soybean variety, Heinong 37

[0018] 2. Strains and plasmids: EHA105-pZY101

[0019] 2. The composition of the basic medium and the composition of the medium at different stages

[0020] Table 1MS medium composition

[0021]

[0022] Table 2 The formula of different stages of soybean tissue culture medium

[0023]

[0024] Note: The above dosages are for 1L volume of medium. The basic components of the liquid co-medium and solid co-medium are the same as the bud induction medium, except that the amount of macronutrients is reduced by 1 / 10, and the pH drops to 5.4; in addition, 40mg of acetosyringone should be added to the liquid co-medium, solid The co-culture medium was added with 40 mg of acetosyringone, 400 mg of L-cysteine, dithiothreitol, 154 mg, and 158 mg of sodium thiosulfate. Add 0...

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Abstract

The invention discloses a soybean genetic transformation method which is implemented by carrying out agrobacterium-mediated regeneration and genetic transformation culture by utilizing a soybean cotyledonary node as an explant. The method mainly comprises the following steps: 1. culturing a soybean seed in a germination culture medium with TDZ (thidiazuron), thereby obtaining a cotyledonary node explant; 2. carrying out genetic transformation on the cotyledonary node explant in an agrobacterium-mediated mode; and 3. obtaining a regenerated transgenic plant by using a special regeneration system. The invention provides a new, quick and efficient soybean tissue regeneration and genetic transformation method which has the advantages of simple culture process and short regeneration time.

Description

Technical field [0001] The invention relates to a soybean genetic transformation method, in particular to a method for genetic transformation of soybean cotyledon nodes and efficient regeneration. Background technique [0002] Soybean (Glycine max (L.) Merrill.) is a diploid (2=40) plant belonging to the Leguminosae, Papilionaceae, and Glycine genus. It is widely planted in various countries in the world and is an important food and economic crop in the world. It is also the main source of edible oil and vegetable protein. Due to various diseases, pests and various non-biological adversities, the quality and yield of soybeans are difficult to meet people's needs. It is of great significance to improve soybean crops through genetic engineering. Although genetically modified soybeans have been planted on a large scale globally, soybean genetic transformation has always been the main obstacle to restricting soybean genetically modified research. The establishment of soybean regener...

Claims

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Application Information

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Patent Type & AuthorityApplications(China)
IPC IPC(8): C12N15/82
Inventor王光霞李国婧万永青王瑞刚
OwnerINNER MONGOLIA AGRICULTURAL UNIVERSITY