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Polygonatum sibiricum endophyte and application thereof

A technology of endophytic bacteria and Polygonatum, applied in endophytic bacteria and its application fields to achieve high antibacterial activity

Active Publication Date: 2018-11-16
HUANGSHAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, there are only a small number of literature reports on the research on Polygonatum endophytes, such as Li Yanling et al. reported the antibacterial activity of Taishan Polygonatum endophytes and Wang Ying et al. reported the antibacterial metabolites of Polygonatum multiflora endophytes; There is no literature report on the study of endophytic bacteria in Anhui Polygonatum

Method used

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  • Polygonatum sibiricum endophyte and application thereof
  • Polygonatum sibiricum endophyte and application thereof
  • Polygonatum sibiricum endophyte and application thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0028] Embodiment 1: Isolation and identification of bacterial strain of the present invention

[0029] A Polygonatum endophyte, the strain is isolated from the rhizome of wild Polygonatum in Guniujiang Nature Reserve in Anhui Province, a group of strains are fermented through various media, and the fermentation product is extracted with ethyl acetate, and the paper diffusion method is used for the fermentation Antibacterial activity analysis, and finally screened and isolated a strain of Polygonatum endophyte. The specific isolation and purification of Polygonatum endophyte refers to the Schulz method and is improved. The overview is as follows: Take fresh rhizomes, wash them with tap water, and take rhizomes with a diameter of about 5mm Treat with 5% (v / v) sodium hypochlorite solution for 5 minutes; rinse with sterile water for 4 times and absorb the residual sterile water with sterile filter paper; then treat the rhizome with 75% (v / v) ethanol solution for 5 minutes; Rinse ...

Embodiment 2

[0034] Embodiment 2: Preparation of endophyte fermentation product

[0035] Take the preserved HJ-1 slant strains, pick a small amount of strains, streak them on fresh PDA medium, and activate them at a constant temperature of 28°C for 3-5 days. The activated single colony was transferred to freshly prepared PD medium, and cultured at 180rpm in a constant temperature shaker at 28°C for 12h to obtain a seed solution. Pipette 10mL of seed solution into 400mL freshly prepared medium of ME, PD, Gaoshi No. 1, modified Chase and MMB, culture in a constant temperature shaker at 28°C at 180rpm for 7 days, and observe the production of metabolites. The fermentation method of each pure strain colony in embodiment 1 is the same as above.

[0036] Take the culture broth after 7 days of fermentation, filter it with 6-8 layers of gauze to remove the bacteria, and then filter it with medium-speed filter paper, and add ethyl acetate to extract according to the ratio of fermentation broth:eth...

Embodiment 3

[0042] Embodiment 3: the bacteriostasis property of fermented product

[0043] The above-mentioned extracted fermentation products were diluted to 3.00mg / mL respectively, and their antibacterial activities against 7 kinds of tested bacteria were detected, and the results were as follows: figure 2 shown. From figure 2 The detection result shows that the fermentation product that this endophyte produces in ME, MMB and PD medium is to Pseudomonas aeruginosa ( figure 2 A), Salmonella typhimurium ( figure 2 B) and Bacillus thuringiensis ( figure 2 The antibacterial effect of C) is very obvious; while the fermentation products produced in Gaoshi No. 1 and Chase medium have no obvious antibacterial activity on the above-mentioned 7 kinds of tested bacteria (not shown). Therefore, the fermentation products produced by ME, MMB and PD medium were selected to further explore their inhibitory concentration.

[0044] The fermentation products produced by ME, MMB and PD medium wer...

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Abstract

The invention discloses a polygonatum sibiricum endophyte and an application thereof. The polygonatum sibiricum endophyte is named HJ-1 and preserved at CCTCC (China Center For Type Culture Collection) in Wuhan University, Wuhan, China on March 28th, 2018, and the preservation number is CCTCC No:M2018159. Research proves that a fermentation product of the endophyte has the function of resisting Pseudomonas aeruginosa, Salmonella typhi and Bacillus thuringiensis.

Description

technical field [0001] The invention relates to the field of bioengineering, in particular to a Polygonatum endophyte and its application. Background technique [0002] Polygonatum sibiricum (Polygonatum sibiricum), also known as chicken claw ginseng, chicken head Polygonatum, etc., is a perennial herb of the genus Polygonatum Mill in the Liliaceae; Polygonatum sibiricum is a traditional Chinese herbal medicine in my country. Its rhizome is used as medicine. Invigorating the spleen and other effects. Polygonatum genus medicinal plant resources are widely distributed in my country, and about 20 species can be used as medicine. Polygonatum multiflora distributed in my country includes Polygonatum long-stemmed, Polygonatum multiflora, Polygonatum multiflora, Polygonatum multiflora, etc. Among them, Polygonatum multiflora has the best quality and efficacy. Polygonatum multiflora is mainly produced in Sichuan, Hunan, Hubei, Anhui and other provinces. Among them, there are more t...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12P1/04A61P31/04C12R1/07
CPCA61P31/04C12N1/20C12P1/04C12N1/205C12R2001/07
Inventor 柏晓辉翟大才方建新胡长玉刘孝莲刘娣汤敏黄松吕顺清潘健
Owner HUANGSHAN UNIV
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