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Method for converting and synthesizing saponin through adventitious root system culture of Chinese thorowax root

A culture medium formula, the technology of Bupleurum, applied in the directions of chemical instruments and methods, horticultural methods, steroids, etc., can solve the problems of long cultivation period, high cost, complicated induction process, etc., and achieve convenient updating, short production period, Simple effects of biotransformation systems

Inactive Publication Date: 2010-11-17
SHANXI UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In terms of in vitro culture of Bupleurum root, there is only a report on the culture of Mishima Bupleurum hairy root, which is obtained through Agrobacterium rhizogenes infection or transgenic technology, the induction process is complicated, the culture period is long, and the cost is high

Method used

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  • Method for converting and synthesizing saponin through adventitious root system culture of Chinese thorowax root
  • Method for converting and synthesizing saponin through adventitious root system culture of Chinese thorowax root
  • Method for converting and synthesizing saponin through adventitious root system culture of Chinese thorowax root

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] The cut section of Bupleurum root taken from Taigu, Shanxi (originally produced in Zuoquan, Shanxi) was used as an explant, washed away the floating soil with running water, cut into 2cm-3cm long section, soaked in 75% ethanol for 30s, 0.1 %HgCl 2 Disinfect the aqueous solution for 10 minutes, rinse with sterile water four times, inoculate in B supplemented with 2,4-D 1.0mg / L, KT 0.2mg / L, sucrose 3.0%, agar 0.7%, pH 5.8 5 culture medium (A medium), cultured at 25±2°C in the dark, and obtained callus after 4w (such as figure 1 ). Subculture once every 30 days, and obtain a large amount of light yellow callus after subculturing three times.

[0030] The pale yellow callus was transferred to rooting induction medium (B medium). The composition of the rooting induction medium is: B 5 Medium + 1.0mg / L calcium diphenylurea disulfonate + 3.0% sucrose + 0.7% agar, pH 5.8. After 35 days, adventitious roots (such as figure 2 ).

[0031] After rooting for 40 days, select ro...

Embodiment 2

[0035] Cut the roots of Bupleurum bupleurum taken from the field of Lingchuan, Shanxi, wash away the floating soil, cut into 2cm-3cm long sections, soak in 75% ethanol for 30s, 0.1% HgCl 2 The aqueous solution was sterilized for 10 min, washed with sterile water four times, inoculated on the B5 medium (A medium) containing 2,4-D 1.0 mg / L, KT 0.2 mg / L, sucrose 3.0%, pH 5.8 (A medium) at 25 Cultured at ±2°C in the dark, the explants dedifferentiated to form callus after 2w-4w, and a large number of callus were obtained after three subcultures.

[0036] Select the compact, light yellow callus and transfer it to the rooting induction medium (B medium) obtained by screening. The composition of B medium is: B 5 Medium + 1.0 mg / L calcium diphenylurea disulfonate + 3.0% sucrose + 0.7% agar, pH 5.8. During the subculture process, adventitious roots continuously differentiated into new roots.

[0037] Every 30d is one generation, and a large amount of adventitious root systems of Bup...

Embodiment 3

[0041] The cut sections of Bupleurum chinensis roots taken from the cultivation site in Ruicheng, Shanxi were sterilized and inoculated into the culture medium to induce callus formation, induce adventitious root differentiation, conduct adventitious root proliferation and adventitious root system formation culture, and conduct biotransformation of active ingredients in adventitious roots synthesis. The culture conditions, culture medium and the extraction and determination of saikosaponin are the same as in Example 1. It is determined that the monthly proliferation rate of adventitious roots reaches 7.212g, the monthly dry weight multiplication multiple reaches 6.67, and the total amount of saikosaponins a and d reaches 0.127%.

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Abstract

The invention provides a method for converting and synthesizing saponin through adventitious root system culture of Chinese thorowax root. The method comprises the following steps of: sterilization of an explant; induction and proliferation of callus; differentiation and induction of an adventitious root; proliferation of the adventitious root and forming of an adventitious root system; and biosynthesis and conversion of saikosaponin. The invention is characterized by selecting the undercutting section of honewort or bupleurum scorzonerifolium willd as the explant and screening the culture medium compositions for inducing the callus, the induced conditions of the adventitious root, the culture medium compositions of adventitious root proliferation and adventitious root system forming and a culture medium for converting and synthesizing the saikosaponin. The culture medium has good acceleration effect on the adventitious root proliferation, the adventitious root system forming and the saponin synthesis. The maximum monthly proliferation ratio of the adventitious root can reach 10.554, the maximum monthly dry weight proliferation multiple thereof can reach 10.352, and the maximum total amount of the saikosaponin a and d can reach 0.149 percent.

Description

technical field [0001] The invention relates to plant tissue culture and biotransformation, in particular to a method for the induction of adventitious roots of a medicinal plant Bupleurum bupleuri, the formation of adventitious roots and its biotransformation to synthesize saikosaponin. Background technique [0002] Bupleurum bupleurum is a perennial herb, and it is also a commonly used bulk Chinese herbal medicine. The medicinal value is very high, and the single component of Bupleurum bupleurum has definite curative effect, among which saikosaponin is the most important medicinal component and quality detection index of medicinal material. [0003] Chinese Pharmacopoeia (2005 Edition) included Bupleurum Chinese DC. (also known as hard seedling Bupleurum) and Bupleurum scorzonerifilium Willd. (also known as soft seedling Bupleurum Chinese DC.) as medicinal The original plant of Bupleurum, the root of Bupleurum and Bupleurum narrow leaves are the authentic products of Bupl...

Claims

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Application Information

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IPC IPC(8): C07J75/00C07J71/00A01H4/00
Inventor 郝建平关倩杨东方韩丹
Owner SHANXI UNIV
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