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

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 ...

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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.

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