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Method for inducing production of tripterygium wilfordii hairy root by agrobacterium rhizogenes

A technology of Agrobacterium rhizogenes and hairy roots, which is applied in the field of using Agrobacterium rhizogenes to induce hairy roots of Tripterygium wilfordii

Inactive Publication Date: 2012-01-18
FUJIAN AGRI & FORESTRY UNIV
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  • Abstract
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Problems solved by technology

[0006] At present, there is no report about utilizing the hairy root of Tripterygium wilfordii to produce its active substances both at home and abroad.

Method used

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  • Method for inducing production of tripterygium wilfordii hairy root by agrobacterium rhizogenes
  • Method for inducing production of tripterygium wilfordii hairy root by agrobacterium rhizogenes
  • Method for inducing production of tripterygium wilfordii hairy root by agrobacterium rhizogenes

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Embodiment Construction

[0015] A method of using Agrobacterium rhizogenes to induce hairy roots of Tripterygium wilfordii of the present invention, the specific content and implementation are as follows:

[0016] 1. Acquisition of aseptic explants of Tripterygium wilfordii: Pick leaves and tender stems of wild tripterygium wilfordii seedlings, rinse with clean water, cut into sections on an ultra-clean workbench, and scratch. Leaves cut into 1cm 2 , The tender stem is 2-4 cm long. Scratch the tender stems 2 to 3 times, and keep the cuts at both ends smooth. The explants were sterilized with 70% alcohol for 1 min, washed with sterile water for 3 times, and then soaked in 0.1% HgCl 2 After sterilizing in the solution for 10 minutes, rinse with sterile water for 3 times, dry the surface moisture with sterile filter paper, place in 1∕2MS liquid medium, and cultivate under 2000lx, 16h / d sunlight, and 25°C for 2-3 days. In order to make the wound site secrete more phenolic substances and improve the con...

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Abstract

The invention provides a method for inducing the production of tripterygium wilfordii hairy roots by agrobacterium rhizogenes, which comprises the following steps: (1) obtaining aseptic explants; (2) preparing bacterial liquid of agrobacterium rhizogenes; (3) performing agrobacterium rhizogenes infection to induce hairy roots; (4) establishing a hairy root in-vitro culture system, wherein the establishment of the hairy root in-vitro culture system comprises the following steps: culturing hairy roots in a 1 / 2 MS solid medium containing 500 mg / L cefotaxime sodium under a dark culture condition with a temperature of 27 + / -1 DEG C, performing subculture once every 7 days, reducing the using concentration of cefotaxime sodium gradually during subculture till no bacterium is found, finally putting the completely aseptic hairy roots on the 1 / 2 MS solid medium for subculture and preservation. During the detection of the produced aseptic tripterygium wilfordii hairy roots, a considerable amount of triptolide is found to be contained in the hairy roots, and the content is significantly higher than the content of triptolide in traditional tripterygium wilfordii wild roots and tissue culture roots; the method has good prospects for industrial application. The production of triptolide by hairy root culture fills the gap of traditional Chinese medicine.

Description

technical field [0001] The invention discloses a method for inducing tripterygium wilfordii Hok . f1 hairy roots by using Agrobacterium rhizogenes, and initially establishes a tripterygium wilfordii Hok . f1 in vitro culture system. Background technique [0002] Agrobaeterium rhizogenes (Agrobaeterium rhizogenes) is a Gram-negative soil Agrobacterium belonging to the genus Agrobacterium (Agrobactefium) of the family Rhizobiaceae (Rhi-zobiaceae), which can infect most dicotyledonous plants and a few monocotyledonous plants. Agrobacterium infects plants, transfers and integrates a piece of DNA (T-DNA) on the Ri plasmid it contains into the plant genome, and induces hairy roots. [0003] Tripterygium wilfordii is a dicotyledonous woody vine, distributed in southern Fujian, Jiangxi, Zhejiang, Guangdong, Hunan, Hubei, Taiwan and other provinces. It is a precious medicinal plant. Tripterygium wilfordii extract contains more than 70 active ingredients, mainly including secondary ...

Claims

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Application Information

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IPC IPC(8): C12N15/82A01H5/06
Inventor 封磊宋萍洪伟吴承祯陈敏捷
Owner FUJIAN AGRI & FORESTRY UNIV
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