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Method for improving yield of salvianolic acid B in savia miltiorrhiza suspension culture cells by inducing

A technique for culturing cells in suspension and salvianolic acid, which is applied in horticultural methods, botanical equipment and methods, horticulture and other directions to achieve the effects of avoiding low efficiency, short induction time and simple process.

Inactive Publication Date: 2012-05-02
NORTHWEST A & F UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This method overcomes the problem of not producing salvianolic acid B after being induced by adding yeast extract, oligogalacturonic acid and other elicitors, and the content of salvianolic acid B in cultured cells of Salvia miltiorrhiza can be increased by about 5 times by using this method

Method used

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  • Method for improving yield of salvianolic acid B in savia miltiorrhiza suspension culture cells by inducing
  • Method for improving yield of salvianolic acid B in savia miltiorrhiza suspension culture cells by inducing
  • Method for improving yield of salvianolic acid B in savia miltiorrhiza suspension culture cells by inducing

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Experimental program
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Embodiment 1

[0045] Fresh Salvia miltiorrhiza seeds were induced to grow sterile seedlings on MS solid medium. The leaves of robust sterile seedlings were selected, cut into small pieces of 0.5 cm × 0.5 cm, and inoculated on MS solid induction medium (containing 1.0 mg / L NAA, 0.5 mg / L 6-BA, 0.5 mg / L 2,4-D, 5.5 g / L agar, 30 g / L sucrose) to induce callus formation. The calli with stable morphological characteristics and growth rate were cultured for one month under the culture conditions of culture temperature of 26°C, light time of 12-16 h, and light intensity of 2000-3000 Lx. Select fresh and loose solid callus, disperse under sterile conditions, transfer the dispersed callus to MS liquid medium (without containing agar and growth regulators, with 30 g / L sucrose), in dark suspension cultures in a shaker. Suspension culture conditions: culture temperature was 25°C, rotation speed was 125 rpm, cultured for 5 days. Add salicylic acid elicitor at a final concentration of 10 mg / L to the medi...

Embodiment 2

[0047] Fresh Salvia miltiorrhiza seeds were induced to grow sterile seedlings on MS solid medium. The leaves of robust sterile seedlings were selected, cut into small pieces of 0.5 cm × 0.5 cm, and inoculated on MS solid induction medium (containing 1.5 mg / L NAA, 0.5 mg / L 6-BA, 0.5 mg / L 2,4-D, 5.5 g / L agar, 30 g / L sucrose) to induce callus formation. The calli with stable morphological characteristics and growth rate were cultured for 1 month under the conditions of culture temperature of 24 °C, light time of 12-16 h, and light intensity of 2000-3000 Lx. Select fresh and loose solid callus, disperse under sterile conditions, transfer the dispersed callus to MS liquid medium (without containing agar and growth regulators, with 30 g / L sucrose), in dark suspension cultures in a shaker. Suspension culture conditions: culture temperature was 25°C, rotation speed was 125 rpm, cultured for 8 days. Add salicylic acid elicitor at a final concentration of 12.5 mg / L to the medium for ...

Embodiment 3

[0049] Fresh Salvia miltiorrhiza seeds were induced to grow sterile seedlings on MS solid medium. The leaves of robust sterile seedlings were selected, cut into small pieces of 0.5 cm × 0.5 cm, and inoculated on MS solid induction medium (containing 1.0 mg / L NAA, 1.5 mg / L 6-BA, 0.5 mg / L 2,4-D, 5.5 g / L agar, 30 g / L sucrose) to induce callus formation. The calli with stable morphological characteristics and growth rate were cultured for 1 month under the culture conditions of culture temperature of 27 °C, light time of 12-16 h, and light intensity of 2000-3000 Lx. Select fresh and loose solid callus, disperse under sterile conditions, transfer the dispersed callus to MS liquid medium (without containing agar and growth regulators, with 30 g / L sucrose), in dark suspension cultures in a shaker. Suspension culture conditions: culture temperature was 25°C, rotation speed was 125 rpm, cultured for 4 days. The salicylic acid elicitor at a final concentration of 6.25 mg / L was added ...

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Abstract

The invention discloses a method for improving yield of salvianolic acid B in savia miltiorrhiza suspension culture cells by inducing, specifically comprising the following steps: 1) cultivating the aseptic seedlings; 2) inducing the callus; 3) sub-cultivating the callus; 4) cultivating the callus by suspending; 5) inducing to produce the salvianolic acid B; and 6) harvesting the culture cells. The method solves the problem that the salvianolic acid B is not produced after the inducing factors, such as yeast extract, galacturonic acid, mycelia extract and the like are added. The content of the salvianolic acid B in the savia miltiorrhiza suspension culture cells is increased by about 5 times by using the method. The technology condition provided by the invention is suitable for industrial production of the salvianolic acid B on a large scale by a cell culture method after being expanded.

Description

technical field [0001] The invention belongs to the technical field of producing natural products by cell culture method, and specifically relates to a method for inducing and increasing the production of salvianolic acid B in salvia miltiorrhiza suspension culture cells. Background technique [0002] Salvia ( Salvia miltiorrhiza The root of bunge is a traditional Chinese herbal medicine. It is mainly used clinically to treat chest and abdomen tingling pain, heat arthralgia pain, sore swelling and pain, restlessness and insomnia, hepatosplenomegaly, angina pectoris and other diseases. The secondary metabolites in Salvia miltiorrhiza——phenolic acid compounds have antithrombotic effects and have protective effects on brain injury caused by cerebral ischemia-reperfusion in mice (Chen et al., 2000), among which salvianolic acid B has a strong scavenging free radicals and antioxidant effects (Huang Yisen et al., 1992). Phenolic acids, represented by salvianolic acid B, are cons...

Claims

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Application Information

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IPC IPC(8): A01H4/00
Inventor 董娟娥梁宗锁碗国伟陈红艳焦蒙丽
Owner NORTHWEST A & F UNIV
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