Method for cultivating transgenic plant

A transgenic plant, plant technology, applied in the fields of botany equipment and methods, biochemical equipment and methods, genetic engineering, etc., can solve problems such as limited salt tolerance, and achieve the effect of improving the possibility

Inactive Publication Date: 2010-06-16
INST OF GENETICS & DEVELOPMENTAL BIOLOGY CHINESE ACAD OF SCI
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Problems solved by technology

[0006] Although it is currently possible to increase the salt tolerance of plants by overexpressing some salt-related genes, it is limited to simply transform a gene to improve the salt tolerance of plants

Method used

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  • Method for cultivating transgenic plant
  • Method for cultivating transgenic plant
  • Method for cultivating transgenic plant

Examples

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Embodiment 1

[0018] Embodiment 1, Agrobacterium-mediated Arabidopsis transformation

[0019] 1) Construction of the binary bacterial artificial chromosome library of Thellungiella halophila

[0020] Extract the total DNA of the aerial part of the small salt mustard grown for 20 days, incompletely digest the total DNA with the restriction endonuclease BamHI, analyze by pulse gel electrophoresis, and extract the DNA fragments with a molecular weight between 50-100kb from the gel The recovered DNA fragments were combined with the plant dual-source expression vector BIBAC2 (Hamilton CM, Frary A, Lewis C, Tanksley, S D (1996) Stable transfer of intact high molecular weight DNA into plant chromosomes. Proc Natl AcadSci US A. 93:9975-9) for the ligation reaction. The ligation product was transformed into Escherichia coli DH10B by electroporation for library amplification. The conversion efficiency of 1 microliter ligation product is 1×10 5 BAC clones, the entire library consisted of 23,040 clo...

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Abstract

The invention discloses a method for cultivating transgenic plant. Recombined binary bacteria artificial chromosome containing foreign DNA is introduced into agrobacterium, so as to obtain recombined agrobacterium; the obtained recombined agrobacterium is subject to suspension by the following solution and then converted into the target plant: solvent is water, solute is non-ionic surfactant nonyl phenol polyoxyethylene ether or Sillwet L-77 and cane sugar; wherein the volume percent concentration of the nonyl phenol polyoxyethylene ether or Sillwet L-77 is 0.05-0.1%, and mass percent concentration of the cane sugar is 5%. By utilizing the method of the invention, foreign DNA segment as high as 75kb can be introduced into the target plant, thus the possibility that gene cluster related toinversion resistance (especially related to salt tolerance) is transformed into the target plant is greatly improved, thereby laying the foundation for cultivating salt tolerant plant.

Description

technical field [0001] The invention relates to a method for cultivating transgenic plants. Background technique [0002] Using soil Agrobacterium to transfer exogenous genes is an effective means in plant genetic engineering. Agrobacterium is a Gram-negative bacterium ubiquitous in soil, which can chemotactically infect the wounded parts of most dicotyledonous plants under natural conditions, and induce the production of corona tumors or hairy roots. The cells of Agrobacterium tumefaciens and Agrobacterium rhizogenes contain Ti plasmid and Ri plasmid respectively, and there is a segment of T-DNA on it. After Agrobacterium enters the cell by infecting the plant wound, T-DNA can be inserted into the plant genome. Therefore, Agrobacterium is a natural plant genetic transformation system. People insert the target gene into the modified T-DNA region, and realize the transfer and integration of the exogenous gene to the plant cells by means of Agrobacterium infection, and then ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/29C12N15/82A01H1/00
Inventor 谢旗夏然张译月
Owner INST OF GENETICS & DEVELOPMENTAL BIOLOGY CHINESE ACAD OF SCI
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