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
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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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