Salvia miltiorrhiza bunge endophyte having phenolic acid accumulation inducing effect, and uses thereof

A technology of endophytic bacteria and salvia miltiorrhiza, applied in the field of endophytic bacteria in natural medicinal plants, can solve the problems of unstable quality and low content of active ingredients, and achieve the effect of increasing the content of phenolic acids in the hairy root of 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

[0011] The technical problem to be solved in the present invention is to provide an endophyte of Salvia miltiorrhiza---Pseudomonas chloropinus, which

Method used

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  • Salvia miltiorrhiza bunge endophyte having phenolic acid accumulation inducing effect, and uses thereof
  • Salvia miltiorrhiza bunge endophyte having phenolic acid accumulation inducing effect, and uses thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Embodiment 1, the isolation culture method of Pseudomonas chloropinus CCTCC NO:M 2015082, carries out the following steps successively:

[0030] 1. The healthy Salvia miltiorrhiza plants were collected and planted in Zhongjiang, Sichuan, their roots were fully washed with tap water, and treated with 25KHz ultrasonic waves for 5 minutes;

[0031] 2. Under aseptic conditions, wash the test material obtained in step 1 with sterile water for 2 times, then use 75% (v / v) alcohol and 3% (m / V) sodium hypochlorite solution to test the test material successively. Carry out surface disinfection to kill microorganisms on the surface of the test material;

[0032] 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 produced in the last rinse and inoculate it on the basic NA medium, at 30°C After culturing for 24 hours, observe whether there are colonies. Based on this, verify whether the abo...

Embodiment 2

[0040] Example 2, the preparation method of the elicitor of Pseudomonas chloropinus CCTCC M 2015082, the following steps are carried out in sequence:

[0041] 1. Transfer Pseudomonas chloropinus CCTCC NO:M 2015082 to the slant of the basic NA medium, culture at 30°C for 2 days; obtain slant seeds;

[0042] Basic NA medium slant formula: 10g peptone, 3g beef extract, 5g NaCl, 18g agar and 1000mL distilled water, pH=7.2~7.4.

[0043] 2. Use an inoculation loop to pick up a loop of the slant bacteria obtained in step 1, inoculate it into a 250ml Erlenmeyer flask containing 100ml NA liquid medium, and culture it at 28°C for 72h with shaking at a speed of 220r / m to obtain Pseudomonas chlorophyll Bacteria CCTCC M 2015082 fermentation stock solution;

[0044] The formula of NA liquid medium is: peptone 10g, beef extract 3g, NaCl 5g and distilled water 1000mL, pH=7.2~7.4.

[0045] The above step 1 and step 2 are all cultured under natural light conditions.

[0046] 3. Take the fermen...

Embodiment 3

[0047] Example 3, the promotion experiment of Pseudomonas chloropinus CCTCC M 2015082 on the accumulation of phenolic acids in the hairy root of Salvia miltiorrhiza:

[0048] 1. 0.3g of Salvia miltiorrhiza hairy root (mother root) was cultured in 100ml of 6,7-V medium at 25°C and 110rpm in the dark for 18 days; as a base;

[0049] Experimental group: Add 1.5ml of Pseudomonas chloropinus CCTCC inducer (obtained in Example 2) to each base, and continue to cultivate under the same conditions (darkness, 25°C, 110rpm) for 6 days; as the experimental group;

[0050] Blank control group: add 1.5ml of sterile NA liquid medium to each base, and continue to cultivate under the same conditions (dark, 25°C, 110rpm) for 6 days; as a blank control group;

[0051] The above-mentioned experimental group and the blank control group were respectively set with 5 repetitions;

[0052] The hairy roots of Salvia miltiorrhiza obtained by the experimental group and the blank control were carried out...

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Abstract

The present invention relates to a nature medicinal plant endophyte salvia miltiorrhiza bunge endophyte, and uses thereof, specifically to a salvia miltiorrhiza bunge endophyte, which is Pseudomonas chlororaphis LG1, and has the preservation number of CCTCC NO:M 2015082. The present invention further discloses the uses of the salvia miltiorrhiza bunge endophyte, wherein the salvia miltiorrhiza bunge endophyte can promote accumulation of phenolic acid substances in salvia miltiorrhiza bunge hairy roots. With the salvia miltiorrhiza bunge endophyte, the problems of low active component content and instable quality during the salvia miltiorrhiza bunge hairy root culture process can be solved.

Description

technical field [0001] The invention relates to a natural medicinal plant endophytic bacterium-endophytic bacterium of salvia miltiorrhiza and its application. Background technique [0002] Salvia miltiorrhiza Bunge is a plant of Labiatae, Salvia genus, dicotyledonous, perennial erect herb. It is a traditional Chinese medicinal material in my country, and its roots and stems are used as medicine. Danshen has a wide range of uses. According to the 2010 edition of "Pharmacopia of the People's Republic of China", Danshen can be used to treat blood circulation, relieve pain, clear heart and troubles, and treat amenorrhea dysmenorrhea, irregular menstruation, heat arthralgia, swelling and pain, upset and insomnia, angina pectoris, etc. . Modern medical research has found that Salvia miltiorrhiza has significant curative effects in dilating coronary blood vessels and improving microcirculation, and is currently mostly used for the treatment of cardiovascular diseases. With the c...

Claims

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Application Information

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IPC IPC(8): C12N1/20A01N63/00A01P21/00C12R1/38
Inventor 胡秀芳杨随娟梁宗锁杨东风李欧杨瑞环尤红苏尚健班善贇沈书立张露
Owner ZHEJIANG SCI-TECH UNIV
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