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Microbacteria and their uses

A microbacteria and application technology, applied in the field of microbacteria and its application, can solve the problems of low activity yield, influence on cultivation amplification and industrialization, etc., and achieve the effect of increasing the content

Active Publication Date: 2017-11-03
ZHEJIANG SCI-TECH UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the currently established hairy root culture system of Salvia miltiorrhiza still has the problem of low activity yield, which affects further scale-up and industrialization of culture.

Method used

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  • Microbacteria and their uses
  • Microbacteria and their uses

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Embodiment 1, Microbacterium arborescens LG2 (CCTCC NO: M 2015083) isolation culture method is as follows:

[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 material obtained in step 1 is rinsed 3 times with sterilized deionized water, and is surface-sterilized with 75% (v / v) alcohol and 3% (m / V) sodium hypochlorite solution respectively, thereby Kill microorganisms on the surface of the test material;

[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 the nutrient agar medium, and incubate at 30°C for 24 hours Afterwards, sterility verification is performed.

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

Embodiment 2

[0033] Embodiment 2, Microbacterium arborescens LG2 (CCTCC NO: M 2015083) elicitor preparation method, the following steps are carried out in sequence:

[0034] 1. Microbacterium arborescens LG2 (CCTCC NO: M 2015083) was transferred to the slant of nutrient agar and cultured at 30°C for 2 days;

[0035] The slant of nutrient agar is: peptone 10g, beef extract 3g, NaCl 5g, agar 18g and distilled water 1000mL, pH=7.2~7.4.

[0036] 2. Use an inoculation loop to pick up a loop of the slant strain obtained in step 1, inoculate it in 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 Microbacterium arborescens LG2CCTCC M2015083;

[0037] The nutrient broth medium is: peptone 10g, beef extract 3g, NaCl 5g and distilled water 1000mL, pH=7.2~7.4.

[0038] The above step 1 and step 2 are all cultured under natural light conditions.

[0039]3. Take the fermentation stock solution obtai...

Embodiment 3

[0041] Embodiment 3, Microbacterium arborescens LG2 (CCTCC NO: M 2015083) promotes the accumulation of rosmarinic acid in Salvia miltiorrhiza root:

[0042] 1. Inoculate 0.3 g of the hairy root (mother root) of Salvia miltiorrhiza into a 250 ml Erlenmeyer flask containing 100 ml of 6.7-V medium, and culture it at 25°C and 100 rpm in the dark for 18 days; as a base;

[0043] Experimental group 1: Add 1.5ml of Microbacterium arborescens LG2 (CCTCCNO: M2015083) supernatant elicitor (gained in Example 2, bacterial liquid elicitor) to each bottle of base material, continue the same conditions (at 25°C, 100rpm dark Cultivation) was cultivated for 6 days; as experimental group 1;

[0044] Experimental group 2: Add 1.5ml of Microbacterium arborescens LG2 (CCTCCNO: M2015083) bacterium elicitor (gained in Example 2) to each bottle of base material, and continue to cultivate under the same conditions (25°C, 100rpm dark culture) for 6 days; as Experimental group 2;

[0045] Negative con...

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Abstract

The invention relates to a microbacterium strain and its application in the salvia miltiorrhiza hairy root culture system. Specifically: the present invention discloses a microbacterium, which is Microbacterium arborescens LG2, and its preservation number is: CCTCC NO: M 2015083. The purpose of the microbacteria is to: promote the accumulation of rosmarinic acid and salvianolic acid B in the hairy root of Salvia miltiorrhiza; and promote the weight gain of the hairy root of Salvia miltiorrhiza.

Description

technical field [0001] The invention relates to a microbacterium strain and its application in the salvia miltiorrhiza hairy root culture system. Background technique [0002] Salvia miltiorrhiza Bunge, also known as red ginseng, purple miltiorrhiza, red root, etc., is a labiatae plant, and its dried roots and rhizomes can be used as medicine. The record of its use as a medicinal material was first seen in "Shen Nong's Materia Medica", and then it was also recorded in the Chinese medicine works of Liang Dynasty, Tang Dynasty, Five Dynasties and Ming Dynasty. It is distributed in Hebei, Shanxi, Shaanxi, Shandong, Henan, Jiangsu, Zhejiang, Anhui, Jiangxi and Hunan, and it is also distributed in Japan. Salvia miltiorrhiza has the effects of promoting blood circulation and regulating menstruation, dispelling blood stasis and relieving pain, cooling blood and eliminating carbuncle, clearing heart-fire and eliminating troubles, nourishing blood and calming the nerves. Salvia roo...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20A01N63/00A01P21/00C12R1/01
CPCA01N63/00C12N1/205C12R2001/01
Inventor 李欧尤红梁宗锁杨东风胡秀芳杨瑞环杨随娟班善贇沈书立张露苏尚健
Owner ZHEJIANG SCI-TECH UNIV
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