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Method for producing melon-like transgene plants by using agrobacterium

A technology of transgenic plants and Agrobacterium, applied in botany equipment and methods, plant products, plant genetic improvement, etc., to reduce pollution, improve transformation efficiency, and increase the chance of transgenic plants

Inactive Publication Date: 2008-09-10
ZHEJIANG ACADEMY OF AGRICULTURE SCIENCES
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  • Abstract
  • Description
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Problems solved by technology

[0003] In order to solve the existing technical problems, the present invention provides a new method, which can reduce the pollution of Agrobacterium after co-culture or infection culture, thereby greatly improving the transformation efficiency and success rate

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  • Method for producing melon-like transgene plants by using agrobacterium
  • Method for producing melon-like transgene plants by using agrobacterium

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Embodiment Construction

[0011] experiment

[0012] In order to further illustrate the effects and implementation modes of the present invention, experiments are now described. These specific implementation examples are only limited enumerations without departing from the spirit of the present invention, and do not exclude other specific implementations produced by those skilled in the art by combining the prior art with the present invention.

[0013] Experiment 1: Infection of watermelon cotyledons with Agrobacterium (10°C)

[0014] 1. Materials

[0015] Medium formula: Configure MS basic solid medium according to the table below (Table 1).

[0016]

[0017] MS basic liquid medium. In contrast to solid media, liquid media do not contain agar. Preparation of YEB minimal medium.

[0018] Weigh 5 g of beef extract (Beef extract), 1 g of yeast extract (Yeast extract), 5 g of polypeptone (Polypeptone), 5 g of sucrose, and 5 g of agar, add water to the volume to 1000 mL, adjust the pH to 7.2, and ...

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Abstract

The invention provides a method for producing melon transgenic plant with agrobacterium, comprising the following steps of infestation and cultivating the agrobacterium with an explant of the melon plant for 5 to 30 minutes at the temperature between 10 and 30 DEG C. In an optimized mode of implementation, the infecting temperature is between 10 and 15 DEG C, and the cultivation period is between 10 and 15 minutes. The method can be used to effectively prevent the agrobacterium from polluting the explant of the plant during the subsequent cultivation, shorten the period for acquiring the transgenic plant, improve the probability for acquiring the transgenic plant, and reduce the manufacturing cost.

Description

technical field [0001] The invention belongs to the field of plant transgenics, specifically, how to prevent the explants from being polluted by the Agrobacterium strains in the subsequent cultivation process when the explants of melon crops are infected with Agrobacterium. Background technique [0002] Use Agrobacterium strains to infect injured plant tissues under certain conditions, and the transformed Agrobacterium strains can insert their own T-DNA or other foreign genes into the genome of the target plant tissue, thereby producing new regenerated plants. This technology is widely used in plant genetic engineering. However, after co-cultivation with Agrobacterium strains and plant tissues, callus and transgenic seedlings must be differentiated after a period of cultivation. During the selection and cultivation of resistant buds after dipping, Agrobacterium contamination will affect plant gene transformation. An important aspect of success rate. Agrobacterium contamina...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/82A01H1/00A01H5/00
Inventor 方丽王汉荣任海英茹水江王连平
Owner ZHEJIANG ACADEMY OF AGRICULTURE SCIENCES
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