Bacillus velezensis XG2 strain for producing phthalide components and application of bacillus velezensis XG2 strain

A bacillus and phthalide technology, applied in the field of microorganisms, can solve the problems of limited number and types of strains, weak function screening, etc., and achieve the effects of low cost, increased plant height, and promotion of germination

Active Publication Date: 2022-02-15
NANJING UNIVERSITY OF TRADITIONAL CHINESE MEDICINE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, some scholars have studied the microbial resources of Angelica sinensis, but the number and types of strains are limited, and the research on functional screening is relatively weak. Further screening out new strains that produce active ingredients of Angelica sinensis and promote the high-quality growth of Angelica sinensis will be of great help to the in-depth development of microbial fermentation production. It is of great significance to find new sources of phthalides, alleviate the shortage of angelica resources, develop biological fertilizers, and carry out green and healthy planting.

Method used

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  • Bacillus velezensis XG2 strain for producing phthalide components and application of bacillus velezensis XG2 strain
  • Bacillus velezensis XG2 strain for producing phthalide components and application of bacillus velezensis XG2 strain
  • Bacillus velezensis XG2 strain for producing phthalide components and application of bacillus velezensis XG2 strain

Examples

Experimental program
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Effect test

Embodiment 1

[0033] A new strain of Bacillus Velez XG2, which was isolated by the following method:

[0034] (1) Oscillation: Strip the soil around the roots of Angelica sinensis, shake off and collect the soil on the rhizosphere surface, put 5g of soil in a 100mL triangular flask of sterile water, oscillate on a constant temperature shaker at 30°C at 160r / min for 2h, let it stand for 30min, and obtain the soil stock solution (10 -1 ), then use a pipette gun to draw 1mL supernatant and add it to 9mL sterile water, and shake well to get (10 -2 ), and so on, to obtain 10 -1 -10 -6 Diluent.

[0035] (2) Separation: draw 0.1mL 10 -4 、10 -5 、10 -6 The soil dilution solution was added to the prepared LB solid medium (composed of 10g / L peptone, 5g / L yeast extract, and 10g / L NaCl), and three replicates were set for each dilution. Invert the plate, incubate at 30°C and observe the growth of the colony at any time, use an inoculation loop to inoculate bacteria with different shapes on the LB ...

Embodiment 2

[0038]A new strain of Bacillus Velez XG2, which is cultivated by the following fermentation method:

[0039] (1) Inoculate 50mL of LB liquid culture medium with a single colony obtained through repeated purification in step (2) of Example 1, activate and cultivate in a constant temperature shaker at 30°C at 160r / min, and take 1mL of activated strains after 24h to fill 500mL of LB liquid The culture medium was fermented in a Erlenmeyer flask, and cultured on a shaker at 30° C. and 160 r / min for 24 hours; after the above fermentation culture, the fermentation broth was collected.

[0040] (2) The LB medium involved in strain isolation and fermentation culture consisted of 10 g / L peptone, 5 g / L yeast extract, and 10 g / L NaCl.

Embodiment 3

[0042] A new strain of Bacillus Velez XG2 was obtained by analyzing and screening the following metabolites:

[0043] (1) Get the fermented liquid sample that obtains in the embodiment 2 and extract three times with equal amount of ethyl acetate, evaporate ethyl acetate, redissolve with methanol, apply ultra-high performance liquid chromatography-triple quadrupole compound linear ion trap mass spectrometry (UPLC -Q-TRAP / MS) combined method for detection, compare the fermentation ion map of the strain with the standard product, and determine whether the fermentation broth contains phthalides.

[0044] (2) The standard substances of six phthalide components ligustilide, n-butylphthalide, butenylphthalide, ligustilide A, H, and I, which are relatively abundant in the root of Angelica sinensis, were mixed for liquid mass analysis. coupled chromatographic analysis, image 3 , 5 , 7, and 9 are the total ion chromatography (Total ion chromatograpy) of the six standard products, and...

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Abstract

The invention relates to a bacillus velezensis XG2 strain for producing phthalide components and application thereof, the strain is obtained by separating, purifying and culturing from rhizosphere soil of angelica sinensis, and is identified as bacillus through microorganism 16S rDNA sequencing. The strain is preserved in the China Center for Type Culture Collection, and the preservation number is CCTCC NO: M 2021766. A bacillus velezensis strain which is stable in character, simple in culture medium and good in growth is provided for large-scale fermentation preparation of phthalide components, a practical, simple and convenient fermentation culture method and a separation and extraction process thereof are established, and components obtained after fermentation liquor is extracted and concentrated can serve as a new source of active components of angelica sinensis. The bacillus velezensis XG2 can promote the germination of angelica sinensis seeds and improve the plant height and fresh weight of angelica sinensis seedlings.

Description

technical field [0001] The invention relates to a strain of bacillus velei, which can produce phthalide components and can promote angelica seed germination and plant growth, and belongs to the technical field of microorganisms. Background technique [0002] The structure and composition of the bacterial community in the soil can effectively reflect the health of the soil ecosystem, the size of the soil productivity, and the level of environmental disturbance. Changes in the population and structure of rhizosphere microorganisms can cause changes in the function of the microbial flora. . Plant growth promoting rhizobacteria (PGPRs) are a group of autogenous bacteria distributed on the surface of plant roots in the soil around the root system, which affect the plant root structure by secreting related plant hormones or hormone precursors. PGPR promotes the secretion of plant hormones such as indole acetic acid, abscisic acid, cytokinin, gibberellin, and ethylene through nitr...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12P17/04C12R1/07
CPCC12N1/20C12P17/04
Inventor 刘培冯伟萌张森严辉段金廒
Owner NANJING UNIVERSITY OF TRADITIONAL CHINESE MEDICINE
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