Methods for efficient induction production of Glycyrrihiza uralensisi Fisch hairy roots and production of licorice root secondary metabolites by using Glycyrrihiza uralensisi Fisch hairy roots

Abstract

The present invention relates to methods for induction production of hairy roots through infection of Glycyrrihiza uralensisi Fisch seedling stem nodes by using Agrobacterium rhizogenes, and production of medicinal secondary metabolites (such as liquiritin, glycyrrhizic acid and the like) by using the hairy roots, particularly to a method for efficient induction production of Glycyrrihiza uralensisi Fisch hairy roots and a method for production of licorice root secondary metabolites by using the hairy roots. The method is characterized by comprising: (1) obtaining sterile explants, wherein Glycyrrihiza uralensisi Fisch seeds are taken, sterilized, and then inoculated in a seedling culture medium to culture to obtain sterile seedlings with a plant age of 12-15 d, and the roots are cut so as to be adopted as the explants; (2) preparing an Agrobacterium rhizogenes engineering bacteria solution; and (3) inducing production of hairy roots through needling stem nodes. According to the present invention, the method for efficient and stable induction production of the Glycyrrihiza uralensisi Fisch hairy roots is provided, and the obtained Glycyrrihiza uralensisi Fisch hairy roots are subjected to suspension culture through selecting the appropriate liquid culture medium and are used for producing the medicinal secondary metabolites (such as liquiritin, glycyrrhizic acid and the like).

Description

technical field [0001] The invention relates to a method for infecting the stem nodes of Glycyrrhizae uralensis seedlings with Agrobacterium rhizogenes to induce hairy roots, and using the hairy roots to produce medicinal secondary metabolites, such as liquiritin and glycyrrhizic acid, etc., especially for high-efficiency induction A method for producing licorice hairy roots and a method for producing licorice secondary metabolites from hairy roots. Background technique [0002] Ural licorice (Glycyrrihiza uralensisi Fisch.) is an important traditional Chinese medicine in my country. It has the functions of clearing away heat and detoxification, moisturizing the lungs and eliminating phlegm, relieving spasms and relieving pain, invigorating qi and benefiting the spleen and stomach, etc., and is known as the "king of all herbs". Irreplaceable. The main medicinal components of licorice include glycyrrhizic acid and liquiritin, among which glycyrrhizic acid is a very precious a...

AI Technical Summary

Problems solved by technology

With the continuous development and utilization of licorice resources, the total demand for licorice in the domestic market is 90,000 to 100,000 tons, but the total amount of licorice in the country is only 25,000 to 30,000 tons. Production

In September 2001, in order to protect the increasingly scarce licorice resources, my country restricted the excavation of wild licorice, which aggravated the shortage of licorice resources. At present, artificial cultivation is still the main way of licorice production. However, especially in the content of active ingredients, there is still a big gap between artificially planted licorice and wild licorice, which cannot meet market demand

However, the currently disclosed induction cultures of Glycyrrhizae uralensis hairy roots all use isolated tissues for genetic transformation, so the induction rate is relatively low, and the obtained hairy roots carry a large number of Agrobacterium, which makes it difficult to eliminate bacteria, which is not conducive to continuing the hairy roots. Root suspension culture has little practical application value and cannot meet production needs

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