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Method for genetic transformation of eucommia ulmoides by injection

A genetic transformation and transgenic technology, applied in the direction of using micro-injection, other methods of inserting foreign genetic materials, biochemical equipment and methods, etc., can solve the problems that have not been clearly reported in the application and restrict the genomics and gene function of Eucommia ulmoides, Achieve high genetic transformation efficiency

Active Publication Date: 2021-03-16
GUIZHOU UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

At present, the genetic transformation system of Agrobacterium injection has been widely used in dicotyledonous plants such as soybean and litchi, but the application in Eucommia has not been clearly reported, which restricts the research on Eucommia genomics and gene function

Method used

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  • Method for genetic transformation of eucommia ulmoides by injection
  • Method for genetic transformation of eucommia ulmoides by injection
  • Method for genetic transformation of eucommia ulmoides by injection

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Embodiment 1

[0039] Embodiment 1 A kind of genetic transformation technology utilizing Agrobacterium to inject Eucommia seedlings

[0040] 1.1 Genetic recipient preparation: The Eucommia ulmoides seedlings germinated three days ago, and the first true leaf between the two cotyledons was removed by surgical forceps as the genetic transformation recipient for future use. See figure 1 shown.

[0041] 1.2 Cultivation of Agrobacterium: Agrobacterium tumefaciens strain LBA4404, the vector pCAMBIA1300 carrying the hygromycin resistance gene Hyg and GUS gene was inoculated into the culture medium containing 100 mg / mL kanamycin (Kan) and 100 mg / mL rifampicin (Rif) On the YEP solid medium, culture in 28℃ incubator for 2 days; pick a single colony and put it into 5ml YEP (containing 100mg / mL Kan and 100mg / mLRif) liquid medium, 28℃, 180rpm shaking culture for 12 hours, take 100ul Add to 100ml YEP (containing 100mg / mLKan and 100mg / mL Rif) liquid medium, shake at 28°C and 180rpm until suitable, and pr...

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Abstract

The invention relates to a method for genetic transformation of eucommia ulmoides by injection. According to the method, eucommia ulmoides seedlings germinating for three days are taken, a first trueleaf between two cotyledons is pulled out to serve as a genetic transformation receptor, and agrobacterium resuspensions with different concentrations are injected between the two cotyledons of each eucommia ulmoides seedling. The method comprises the following specific operation steps: (1) preparing the genetic transformation receptor; (2) culturing agrobacterium; (3) preparing the agrobacteriumresuspensions; (4) performing genetic transformation of the eucommia ulmoides; (5) managing a transgenic plant; and (6) identifying the transgenic plant. According to the method, a tissue culture technology is not adopted, besides agrobacterium culture, sterile operation is not needed, and the method has the advantages of being easy to operate, low in cost, capable of converting large-fragment DNA, stable in heredity, low in copy number and the like, and becomes the most widely applied genetic transformation method at present.

Description

technical field [0001] The technical field of the invention is plant biotechnology, and specifically relates to a method for genetically transforming eucommia by injection. Background technique [0002] Eucommia ulmoides Oliver (Eucommia ulmoides Oliver) is a deciduous tree belonging to Eucommia ulmoides. In recent years, in the genetic transformation technology of Eucommia mediated by Agrobacterium, tissue culture technology is the main method, but there are factors such as low efficiency of callus induction differentiation, rooting of resistant shoots, and low survival rate of transgenic plants transplanted, resulting in Eucommia This type of research is relatively lagging behind, and it is difficult to obtain larger surviving plants. [0003] The Agrobacterium injection method is a non-tissue culture genetic transformation method in which the genetically transformed recipient is injected with an Agrobacterium suspension in an open environment. The genetic transformation...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/89A01H5/00A01H6/00
CPCC12N15/89C12N15/8205
Inventor 赵懿琛王超赵德刚
Owner GUIZHOU UNIV
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