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Endophyte having salvia miltiorrhiza hairy root phenolic acid accumulation inducing effect, and uses thereof

A technology of endophytic bacteria and hairy roots, applied in the field of endophytic bacteria in Salvia miltiorrhiza

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 current research focuses on using fungi or yeast from non-biological elicitors or biological elicitors, and the components that increase the yield are concentrated on tanshinone compounds, and there is no research on the increase in the content of bacterial elicitors and water-soluble phenolic acids.

Method used

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  • Endophyte having salvia miltiorrhiza hairy root phenolic acid accumulation inducing effect, and uses thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Embodiment 1, the isolation culture method of Olivibacter soli LG3 (CCTCC NO:M 2015084), 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, rinse the test material obtained in step 1 with sterilized deionized water for 3 times, and then use 75% (v / v) alcohol and 3% (m / V) sodium hypochlorite solution surface disinfection successively ;

[0026] 3. In a sterile environment, continue to 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. ...

Embodiment 2

[0032] Embodiment 2, the preparation method of Olivibacter soli LG3 (CCTCC NO: M 2015084) bacterium elicitor, carries out the following steps successively:

[0033] 1. Olivibacter soli LG3 (CCTCC NO:M 2015084) was transferred to nutrient agar slant and cultured at 28°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 the slant strain obtained in Step 1 of 1 loop, 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 Olivibacter soli LG3CCTCC M 2015084;

[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 in step 2, centrif...

Embodiment 3

[0039] Example 3, Olivibacter soli LG3 (CCTCC NO: M 2015084) bacterial elicitor promotes the water-soluble components of Salvia miltiorrhiza root:

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

[0041]Experimental group: Add 1.5ml of Olivibacter soli LG3 (CCTCC NO:M 2015084) cell elicitor (obtained in Example 2) to each base, and continue to cultivate under the same conditions (25°C, 100rpm, protected from light) for 6 days; as the experimental group;

[0042] Negative control: Add 1.5ml of sterile deionized water to each base, and continue to culture under the same conditions (25°C, 100rpm, protected from light) for 6 days; as a blank control group;

[0043] The above-mentioned experimental group and the negative control (blank control group) were respectively set up with 5 repetitions.

[0044] The Salvia miltiorrhi...

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Abstract

The present invention relates to salvia miltiorrhiza endophyte and uses thereof, wherein the salvia miltiorrhiza endophyte is used for accumulating phenolic acid substances during salvia miltiorrhiza hairy root culture. Particularly the present invention discloses the endophyte, which is Olivibacter soli LG3 and has the preservation number of CCTCC NO:M2015084. With the Olivibacter soli LG3, the accumulation of the phenolic acid substances (salvianolic acid B and rosmarinic acid) in the salvia miltiorrhiza hairy root can be promoted, and the salvia miltiorrhiza hairy root weight gain can be promoted.

Description

technical field [0001] The invention relates to an endophytic bacterium of Salvia miltiorrhiza and its use, which can improve the accumulation of phenolic acid substances when it is used for the cultivation of hairy roots of Salvia miltiorrhiza. Background technique [0002] Salvia miltiorrhiza (scientific name: Salvia miltiorrhiza) is a plant of the genus Salvia in the Labiatae family, and the root of Salvia miltiorrhiza is a traditional Chinese medicine. Salvia miltiorrhiza first appeared in the classics "Shen Nong's Materia Medica", which listed danshen as a top-grade medicinal material, which means it is non-toxic. The 2010 edition of "Pharmacopoeia of the People's Republic of China" records that Salvia miltiorrhiza has the functions of promoting blood circulation and promoting menstruation, clearing heart and eliminating troubles, dispelling blood stasis and relieving pain, reducing inflammation and nourishing blood, etc. It is widely used in the treatment of cardiovas...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/14A01N63/04A01P21/00C12R1/645
Inventor 李欧尤红梁宗锁杨东风胡秀芳杨随娟杨瑞环张露班善贇苏尚健沈书立
Owner ZHEJIANG SCI-TECH UNIV
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