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Cultivation method for inducing scutellaria baicalensis hairy root based on agrobacterium rhizogenes infection

A technology of Agrobacterium rhizogenes and a cultivation method, applied in the field of artificial cultivation of Scutellaria baicalensis, can solve problems such as shortage of Scutellaria baicalensis resources, and achieve the effects of strong secondary metabolite capacity, low energy consumption, and stable genetic traits

Inactive Publication Date: 2012-11-07
SHAANXI UNIV OF SCI & TECH
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] The problem to be solved by the present invention is to provide a cultivation method for inducing hairy roots of Scutellaria baicalensis based on Agrobacterium rhizogenes infection, which can induce hairy roots quickly and with a high induction rate, and solve the problem of shortage of Scutellaria baicalensis resources

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  • Cultivation method for inducing scutellaria baicalensis hairy root based on agrobacterium rhizogenes infection
  • Cultivation method for inducing scutellaria baicalensis hairy root based on agrobacterium rhizogenes infection
  • Cultivation method for inducing scutellaria baicalensis hairy root based on agrobacterium rhizogenes infection

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Embodiment 1

[0043] A method for cultivating hairy roots of Scutellaria baicalensis based on Agrobacterium rhizogenes infection, comprising the following steps:

[0044] 1) Acquisition and pre-cultivation of Scutellaria baicalensis explants

[0045] After germination, sterile scutellaria baicalensis seeds are inoculated in a pre-culture medium and cultured at 25-28° C. to grow into seedlings, and the stems of the seedlings are cut out as explants.

[0046] Alternatively, the Scutellaria baicalensis seedlings are directly sterilized, and then the stems of the seedlings are cut as explants.

[0047] Inoculate the obtained aseptic Scutellaria baicalensis explants on the pre-culture medium, and pre-cultivate them under dark conditions at 25-28°C for 2-3 days;

[0048] The pre-medium is MS solid medium containing 0.15-1.0 mg / kg 6-BA and 0.1-0.35 mg / kg NAA (specifically, 1 LMS medium supplemented with 30 g of sucrose and 5 g of agar can be used).

[0049] 2) Activation of Agrobacterium rhizoge...

Embodiment 2

[0070] The optimized cultivation of hairy roots is the same as the screening method in Example 1, except that the added components of the optimized medium are different.

[0071] The optimized medium is to add 40g / L glucose, 0.5g / L hydrolyzed milk protein, 50mg / L Aspergillus niger elicitor, 1.0mmol / L Ca on the hairy root liquid basic medium. 2+ And 0.25mg / L of NAA, the pH value is 7.0.

Embodiment 3

[0073] The optimized cultivation of hairy roots is the same as the screening method in Example 1, except that the added components of the optimized medium are different.

[0074] The optimized medium is to add 20g / L of sucrose, 0.45g / L of ammonium nitrate, 40mg / L of Aspergillus oryzae elicitor, 1.0mmol / L of Mg on the hairy root liquid basic medium. 2+ And 0.25mg / L of 6-BA, the pH value is 4.0.

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Abstract

The invention discloses a cultivation method for inducing scutellaria baicalensis hairy root based on agrobacterium rhizogenes infection, which uses an agrobacterium rhizogenes bacteria liquid to infect root of a scutellaria baicalensis explant and induces the scutellaria baicalensis explant to form a hairy root, the scutellaria baicalensis hairy root presents yellow hairy on a MS solid medium, so that scutelloside can be stably synthesized, an ITS sequencing is performed and the hairy root can be identified as the scutellaria baicalensis hairy root of a scutellaria. Then seed selection is performed for further in a seed selection medium and a cultivation condition can be optimized to select lines which raise the content of active ingredient scutelloside used for hairy root traditional Chinese medicines. Hairy root can be obtained through scutellaria of a scutellaria introduced by agrobacterium rhizogenes, wherein the inductivity can reach 10 - 40%, the obtained scutellaria baicalensis hairy root presents yellow hairy which is capable of stably synthesizing scutelloside. According to the invention, the scutellaria baicalensis hairy root of a scutellaria is cultivated on the optimized medium, the growth is rapid and the biomass can be increased for 23 -35 times in 30 - 35 days, and the cultivation is performed under the dark conditions. The invention has the characteristics of low energy consumption and low cost, which is suitable for industrialization production.

Description

technical field [0001] The invention belongs to the technical field of artificial cultivation of Scutellaria baicalensis, and relates to a cultivation method for inducing hairy roots of Scutellaria baicalensis based on Agrobacterium rhizogenes infection. Background technique [0002] Scutellaria baicalensis Georgi. is a plant of the Labiatae family and is a commonly used traditional Chinese medicine. It is cold in nature and bitter in taste, and it belongs to the lung, heart, liver, gallbladder, and large intestine meridian. Scutellaria baicalensis is a perennial herb that likes a mild climate and is resistant to cold and drought. It is mainly distributed in Hebei, Liaoning, Shaanxi, Shandong, Inner Mongolia and other places. The main components are baicalin, baicalein, wogonin, wogonin, etc. With the in-depth research on Scutellaria baicalensis in the world, baicalin, the main component of Scutellaria baicalensis, has good effects in scavenging oxygen free radicals, reduci...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/82C12N5/04A01H5/06
Inventor 齐香君李雅郭乐康陈秀清钱卫东
Owner SHAANXI UNIV OF SCI & TECH
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