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Method for establishing adventitious root cultivation system of Psammosilene tuniceoides W. C. Wu et C. Y. Wu and expanding cultivation method of Psammosilene tuniceoides W. C. Wu et C. Y. Wu

A culture system and technology for expanding culture, which is applied in the field of establishment and expansion of the culture system of Adventitia chinensis, can solve the problems of retention, growth stability, culture conditions and various adjustment parameters, such as the contradiction between cell growth and accumulation of secondary metabolites , to achieve the effect of saving land, rapid and convenient scale production, and reducing production costs

Active Publication Date: 2012-11-14
成都市三禾田生物技术有限公司
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  • Summary
  • Abstract
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AI Technical Summary

Problems solved by technology

[0007] But so far, the plant cell culture of Jintiezi still stays at the level of in vitro culture and rapid propagation research. There is no information about the use of plant cell culture technology to cultivate Jintiezi tissue, the intracellular synthesis pathway of Jintiezi secondary metabolite saponin, and through comprehensive regulation A report on improving the plant cell biomass of Jintiesuo by technology, and then increasing the content of secondary metabolites such as total saponins of Jintiesuo
The main problem is that there is insufficient research on the growth stability, culture conditions and various regulatory parameters, cell growth and the accumulation of secondary metabolites in the plant cell culture of Jintie Suo.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0062] (1) Select the golden iron lock leaf as the explant, and carry out the disinfection treatment: rinse with running water, soak in Tween 20 for 25 minutes, rinse with sterile water 3 times; then use 0.1% HgCl 2 Treat for 8 minutes, rinse with sterile water for 3 times; then treat with 75% ethanol for 8 seconds, rinse with sterile water for 3 times, and finally absorb the water with sterile filter paper;

[0063] (2) Callus induction: inoculate the explants in step (1) in callus induction medium: MS medium + 2,4-D 0.5 mg / L + NAA 0.5 mg / L + KT 0.1 mg / L+cysteine ​​1.0mg / L, pH 6.0, culture in the dark at 25±1℃. The leaves are inoculated on the induction medium for about 10 days to start to grow milky white callus; about 20 days, the callus gradually covers the explants, and at 25 days, select loose, well-growing callus for subculture;

[0064] (3) Induced callus subculture: 0.5g of callus obtained in step (2) was inoculated into MS medium + sucrose 40g / L + KNO 3 1.9 g / L + NH 4 N...

Embodiment 2

[0070] (1) Select the golden iron lock stem segment with a length of 1.0-1.5cm as the explant, and perform disinfection treatment: rinse with running water, soak in Tween 20 for 25 minutes, rinse with sterile water 3 times; then use 0.1% HgCl 2 Treat for 8 minutes, rinse with sterile water for 3 times; then treat with 75% ethanol for 8 seconds, rinse with sterile water for 3 times, and finally absorb the water with sterile filter paper;

[0071] (2) Callus induction: inoculate the explants in step (1) in callus induction medium: MS medium + 2,4-D 0.5 mg / L + NAA 0.5 mg / L + KT 0.1 mg / L+cysteine ​​2.0mg / L, pH 6.0, culture in the dark at 25±1℃. The leaves are inoculated on the induction medium for about 10 days to start to grow milky white callus; about 20 days, the callus gradually covers the explants, and at 25 days, select loose, well-growing callus for subculture;

[0072] (3) Induced callus subculture: 0.5g of callus obtained in step (2) was inoculated into MS medium + sucrose 40...

Embodiment 3

[0079] (1) Select the stem section 2.0-3.0cm in length as the explant, and perform disinfection treatment: rinse with running water, soak in Tween 20 for 25 min, rinse with sterile water 3 times; then use 0.1% HgCl 2 Treat for 8 minutes, rinse with sterile water for 3 times; then treat with 75% ethanol for 8 seconds, rinse with sterile water for 3 times, and finally absorb the water with sterile filter paper;

[0080] (2) Callus induction: inoculate the explants in step (1) in callus induction medium: MS medium + 2,4-D 0.5 mg / L + NAA 0.5 mg / L + KT 0.1 mg / L, pH 6.0, culture at 25±1℃ and natural light. The leaves are inoculated on the induction medium for about 10 days to start to grow milky white callus; about 20 days, the callus gradually covers the explants, and at 25 days, select loose, well-growing callus for subculture;

[0081] (3) Induced callus subculture: 0.5g of callus obtained in step (2) was inoculated into MS medium + sucrose 40g / L + KNO 3 1.9 g / L + NH 4 NO 3 1.65 g / ...

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Abstract

According to the invention, young leaves or stems of plants of Psammosilene tuniceoides W. C. Wu et C. Y. Wu are taken as an explant to successfully induce callus of the Psammosilene tuniceoides W. C. Wu et C. Y. Wu, and a callus culture system of the Psammosilene tuniceoides W. C. Wu et C. Y. Wu under the conditions of light and dark cultivation is established to induce the differentiation of the callus to produce adventitious roots, thus establishing an adventitious root cultivation system of the Psammosilene tuniceoides W. C. Wu et C. Y. Wu. Moreover, the content of total saponins of the Psammosilene tuniceoides W. C. Wu et C. Y. Wu is determined, and the conditions and parameters of plant cell culture are further optimized, thus establishing a high-yield cell culture system of the Psammosilene tuniceoides W. C. Wu et C. Y. Wu. Therefore, the plant cells of the Psammosilene tuniceoides W. C. Wu et C. Y. Wu are cultured in a large scale to produce the adventitious roots which substitute for the original plant of the Psammosilene tuniceoides W. C. Wu et C. Y. Wu to be used as medicine.

Description

technical field [0001] The invention relates to a method for inducing and cultivating adventitious root systems using plant tissue culture technology, in particular to a method for inducing and cultivating adventitious root systems in the form of gold-iron locks and enlarging bioreactors. [0002] Background technique [0003] Psammosilene tuniceoides (Psammosilene tuniceoides W. C. Wu et C. Y. Wu) is a plant of the genus Psammosilene tuniceoides in Caryophyllaceae, and it is a unique medicinal plant in Southwest my country. The root of Jintiesuo is mainly used as medicine, and it is often used clinically to treat rheumatoid arthritis and other diseases. The main chemical components of Jintiesuo are triterpenes, triterpene saponins, cyclic peptides and lactams. Pharmacological tests have shown that total saponins of Jintiesun have obvious analgesic and anti-inflammatory effects. The traditional method of obtaining Chinese herbal medicine is at the cost of collecting and c...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01H4/00
Inventor 柴素真洪汉君张宗申熊小灿冯敏
Owner 成都市三禾田生物技术有限公司
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