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Method for transforming in-vitro soybean growing points with gene gun

A gene gun transformation and growth point technology, applied in the field of bioengineering, can solve the problems of low plant regeneration frequency and low transformation efficiency, and achieve the effects of small impact, improved transformation efficiency, and high plant regeneration frequency.

Inactive Publication Date: 2011-05-25
上海交通大学中原研究院
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  • Abstract
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Problems solved by technology

In addition, the plant regeneration frequency of somatic embryos is also very low, so the transformation efficiency is very low

Method used

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  • Method for transforming in-vitro soybean growing points with gene gun

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Embodiment Construction

[0030] The embodiments of the present invention are described in detail below. This embodiment is implemented on the premise of the technical solution of the present invention, and detailed implementation methods and specific operating procedures are provided, but the protection scope of the present invention is not limited to the following implementation example.

[0031] The soybean seeds in this example come from this laboratory.

[0032] This embodiment includes the following steps:

[0033] Step 1, the chlorine gas that reacts with 75ml sodium hypochlorite and 25ml concentrated hydrochloric acid is used to sterilize the surface of soybean seeds for 1 hour, and then inoculate into the seed germination medium of MS that adds 0.5mg / L 6-benzylaminopurine (i.e. MS+0.5mg / L 6-BA), to obtain sterile vaccine;

[0034] Step 2: Peel off enough growth points with 4-6 leaf primordia and a length of 0.4-0.6 cm, peel off the leaf primordia to expose the growth points, and then put the...

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Abstract

The invention discloses a method for transforming in-vitro soybean growing points with a gene gun, belonging to the technical field of bioengineering. The method is characterized by making bullets with carriers containing target genes, loading the bullets into the gene gun, then bombarding aseptic seedlings and performing inoculation induction to obtain T0-generation plants; performing rooting induction and seedling hardening by opening a vessel lid on the T0-generation plants, and then inducing the plants to bloom and fruit to obtain T1-generation seeds; and finally transplanting the T1-generation seeds to a phytotron or large fields to obtain T2-generation plants and homozygous positive transformant thereof. The method has the following advantage: by utilizing the characteristic that the plants prepared from the in-vitro soybean growing points are relatively easy to regenerate, the transformation efficiency can be obviously improved.

Description

technical field [0001] The invention relates to a method in the field of bioengineering, in particular to a biolistic transformation method for isolated soybean growth points. Background technique [0002] Soybean is the fifth largest crop in the world today, after wheat, rice, corn and barley. my country is not only a big producer of soybeans, but also a big consumer, and imports a large amount of genetically modified soybeans every year. Compared with developed countries, my country's soybean technology level is relatively backward, and the quality of soybeans lacks international competitiveness. Strengthening the independent research and utilization of genetically modified soybeans is the development direction that my country should adhere to in the soybean industry in the future. At present, the main factor that restricts the further development of soybean transgenic technology in my country is the incomplete regeneration system of soybean cotyledon nodes, hypocotyls, ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/82C12N15/87
Inventor 王贵荣王志民代田纯候彩玲徐美红
Owner 上海交通大学中原研究院
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