Centrifugal sedimentation assisted plant transgenic method

A centrifugal sedimentation and transgene technology, applied in the biological field, can solve the problems of increasing the degree of trauma on the surface of the receptor material, and the effect of cells inside the receptor is minimal, so as to improve the efficiency of plant transgene and increase the area of ​​trauma

Inactive Publication Date: 2016-11-23
SHANGHAI JIAOTONG UNIV
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  • Abstract
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Problems solved by technology

In response to this problem, someone proposed the idea of ​​ultrasonic-assisted transformation of Agrobacterium. This treatment can increase the degree of trauma on the surface of the receptor material, but has little effect on the cells inside the receptor.

Method used

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Embodiment Construction

[0016] The embodiments of the present invention are described in detail below: the present embodiment is implemented under the premise of the technical solution of the present invention, and detailed implementation and specific operation process are provided, but the protection scope of the present invention is not limited to the following implementation example.

[0017] Tobacco for the following examples was obtained from the applicant's laboratory.

[0018] This embodiment includes the following steps:

[0019] Step 1, taking tender leaves from sterile tobacco tissue culture seedlings, cutting off the leaf margins as receptor materials;

[0020] Step 2, activate the Agrobacterium TG1 containing the GUS gene to OD value = 0.6, centrifuge at 50,000 rpm for 5 minutes, and then resuspend the gene with MS liquid culture;

[0021] Step 3, soaking the tobacco recipient leaves in step 1 into the Agrobacterium resuspension, and centrifuging at 5000rpm for 10min;

[0022] Step 4, ...

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Abstract

A centrifugal sedimentation-assisted plant transgenic method is as follows: the plant receptor material is mixed with the agrobacterium, and the centrifugal sedimentation treatment is carried out at a reasonable rotational speed, and finally a positive transformant is obtained. Centrifugal sedimentation can not only increase the wound area of ​​the receptor material, but also allow Agrobacterium to penetrate into the interior of the receptor material under the action of centrifugal force, which significantly improves the efficiency of plant transgenesis.

Description

technical field [0001] The invention relates to a biotechnology, in particular to a method for centrifugal sedimentation to assist plant transgene. Background technique [0002] As the core of biotechnology, transgenic technology has great development potential in plant improvement, medicine, animal husbandry and food industry. However, the plant transgenic technology based on Agrobacterium transformation currently has the problem of low transformation efficiency, and the transformation rate of most plants is about 1%. This problem has also become the main factor limiting the industrial production of transgenic plants. Studies have shown that when Agrobacterium soaks recipient plant material, only a small number of wounded cells on the surface are likely to be transformed, while most of the inner cells in the recipient material have no opportunity to come into contact with Agrobacterium. In response to this problem, someone proposed the idea of ​​ultrasonic-assisted transfo...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/84A01H5/00
Inventor 王贵荣李斌连唐克轩
Owner SHANGHAI JIAOTONG UNIV
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