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Salvia miltiorrhiza endophyte with danshinolic acid B accumulation inducing effect, and use thereof

A technology of endophytic bacteria and salvianolic acid, applied in the application, bacteria, biocide and other directions, can solve the problems of low yield of hairy root active ingredients, affecting the expanded culture of hairy roots and even industrial production, etc., and achieve the effect of increasing the content.

Active Publication Date: 2015-08-12
ZHEJIANG SCI-TECH UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the yield of active ingredients in hairy roots is low, which affects the expansion of hairy roots and even industrial production.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Embodiment 1, the isolation culture method of Pseudomonas psychrotolerans LG4 (CCTCC NO:M 2015085), carries out the following steps successively:

[0024] 1. Collect wild Salvia miltiorrhiza plants in Zhongjiang, Sichuan, wash the roots and then ultrasonically treat them at 25KHz for 5 minutes;

[0025] 2. Under aseptic conditions, the test material obtained in step 1 was rinsed 3 times with sterilized deionized water, and sterilized with 75% (v / v) alcohol and 3% (m / V) sodium hypochlorite solution surface respectively ;

[0026] 3. In a sterile environment, rinse the test material obtained in step 2 with sterile water for 3 times, take 100 μl of the sterile water rinse solution from the last rinse and inoculate it on nutrient agar, and incubate at 30°C for 24 hours. Perform sterility verification.

[0027] If no colonies are generated, continue to the subsequent step 4; if colonies are generated, return to step 2 to continue the surface disinfection treatment.

[002...

Embodiment 2

[0032] Example 2, the preparation method of Pseudomonas psychrotolerans LG4 (CCTCC NO: M 2015085) elicitor, the following steps are carried out in sequence:

[0033] 1. Transfer Pseudomonas psychrotolerans LG4 (CCTCC NO:M 2015085) to nutrient agar slant and culture at 30°C for 2 days;

[0034] Nutrient agar slant: peptone 10g, beef extract 3g, NaCl 5g, agar 18g and distilled water 1000mL, pH=7.2~7.4.

[0035] 2. Use an inoculation loop to pick up a loop of the slant strain obtained in step 1, inoculate it into a 250ml Erlenmeyer flask containing 50ml of nutrient broth medium, and culture it at 220rpm at 28°C for 72h to obtain the fermentation stock solution of Pseudomonas psychrotolerans LG4CCTCC M 2015085;

[0036] The formula of nutrient broth medium is: peptone 10g, beef extract 3g, NaCl 5g and distilled water 1000mL, pH=7.2~7.4.

[0037] The above step 1 and step 2 are all cultured under natural light conditions.

[0038] 3. Take the fermentation stock solution obtained ...

Embodiment 3

[0039] Example 3, Pseudomonas psychrotolerans LG4 (CCTCC NO: M 2015085) promotion experiment on salvianolic acid B accumulation in Salvia miltiorrhiza:

[0040]1. 0.3g of Salvia miltiorrhiza hairy root (mother root) was inoculated into a 250ml Erlenmeyer flask containing 100ml of 6,7-V medium, cultured at 25°C, 100rpm in the dark for 18 days, as the base;

[0041] Experimental group: 1.5 ml of Pseudomonas psychrotolerans LG4 (CCTCC NO: M2015085) elicitor (obtained in Example 2) was added to each base, and cultured under the same conditions (25°C, 100 rpm, dark) for 6 days; as the experimental group ;

[0042] Negative control (blank control): add 1.5 ml of sterile nutrient broth medium to each base, and continue to culture under the same conditions (25°C, 100 rpm, dark) for 6 days; negative control;

[0043] The above experimental group and blank control group were set up with 5 repetitions;

[0044] The hairy roots of Salvia miltiorrhiza obtained by the experimental group a...

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PUM

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Abstract

The invention relates to a Salvia miltiorrhiza endophyte and its use in the improvement of danshinolic acid B accumulation in the Salvia miltiorrhiza hairy root culturing process. The Salvia miltiorrhiza endophyte is Pseudomonas psychrotolerans LG4 concretely, and has a preservation number of CCTCC NO:M2015085. The Salvia miltiorrhiza endophytec an promote the accumulation of a phenolic acid substance (danshinolic acid B) in Salvia miltiorrhiza hairy root.

Description

technical field [0001] The invention relates to an endophytic bacterium of salvia miltiorrhiza and its ability to increase the accumulation of salvianolic acid B when it is used for the cultivation of hairy roots of salvia miltiorrhiza. Background technique [0002] Salvia miltiorrhiza is a perennial herb of Lamiaceae, also known as Dahongpao, red root, red ginseng, purple salvia miltiorrhiza, blood ginseng, etc. It is distributed in North China, East China, Central South, Northwest, Southwest and other regions of China. The active ingredients contained in its roots and rhizomes can be divided into the following two categories according to their properties: fat-soluble diterpenoids and water-soluble phenolic acids. Among them, the water-soluble ingredients are: protocatechualdehyde, caffeic acid, isoferulic acid, rosmarinic acid, methyl rosmarinate, shikonic acid B, monomethyl shikonate, dimethyl shikonate, B Esters, salvianolic acid A (danshensu) and salvianolic acid A, B,...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20A01N63/00C12R1/38
CPCA01N63/00C12N1/205C12R2001/38
Inventor 胡秀芳杨瑞环梁宗锁杨东风李欧杨随娟尤红沈书立苏尚健张露班善贇
Owner ZHEJIANG SCI-TECH UNIV
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