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Separation screening method for antibiotic antituberculotic plant endophyte

A technology for separation and screening of plant endophytes, which is applied in the field of isolation and screening of antibacterial and anti-tuberculosis plant endophytes, achieving the effect of simple operation

Inactive Publication Date: 2008-07-23
JIANGSU UNIV OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] The antibacterial and anti-tumor activity of plant endophytes, many scientists have carried out in-depth research, and obtained a series of active monomers, and through the optimized surface disinfection method, the study of plant endophytes such as camphor and angustifolia Antibacterial and anti-tuberculosis activity, through fermentation to obtain secondary metabolites that have inhibitory effects on human and animal pathogens and Mycobacterium tuberculosis, thereby obtaining antibacterial and anti-tuberculosis active strains, which has not received much attention in the field of endophytic bacteria research

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] The tissues of various parts of the ten genus Angustifolia collected from the campus of Chongqing University were selected as the host samples of endophytes, and the surface was disinfected according to the following procedures:

[0026] (1) In the aseptic operating table, the host sample of the plant endophyte is washed 3 times with sterile water;

[0027] (2) Use the disinfection combination of 75% alcohol + 0.1% mercury liter, soak in 75% alcohol for 1-5min (interval 1min), rinse with sterile water 3 times, rinse in 0.1% mercury liter for 0.5-2min (interval 0.5min) min), rinsed with sterile water for 5 times, and collected the sterile water used to rinse the plant tissue for the last time as a control.

[0028] The results showed: Roots and stems were soaked in 75% alcohol for 1 min, washed 3 times with sterile water, rinsed with 0.1% mercury liter for 1.5 min, and 5 times with sterile water; leaves and fruits were soaked in 75% alcohol for 1 min, washed 3 times with...

Embodiment 2

[0030] (1) Through the disinfection scheme of Example 1, the endophytic bacteria grown from the antibacterial and anti-tuberculosis plant tissues were picked and inoculated on the beef extract solid medium according to different colony forms, and kept at a constant temperature of 37°C Cultivation: pick endophytic fungi and inoculate them on the PDA medium by streaking, and culture at a constant temperature of 27°C. After purification, inoculate the corresponding slant and save it for later use;

[0031] (2) Carry out liquid fermentation culture of the above-mentioned endophytic bacterial strains: the endophytic fungus medium is PDA liquid medium, the endophytic bacteria is beef extract liquid medium, the medium is subpackaged in the Erlenmeyer flask, picked with an inoculation loop Take a small amount of purified endophytic bacteria in a culture bottle, put the fungus on a shaker at 27°C, 150r / min, and shake it for 6-7 days, put the bacteria on a shaker at 37°C, 150r / min, shak...

Embodiment 3

[0038] (1) Through the disinfection scheme of Example 1, the endophytic bacteria grown from the medicinal plant tissue were picked and inoculated on the beef extract solid medium according to different colony forms, and cultured at a constant temperature of 37°C ; Pick the endophytic fungi and inoculate them on the PDA medium by streaking, and cultivate them at a constant temperature of 27°C. After purification, inoculate the corresponding slant and save it for later use;

[0039] (2) Carry out liquid fermentation culture of the above-mentioned endophytic bacterial strains: the endophytic fungus medium is PDA liquid medium, the endophytic bacteria is beef extract liquid medium, the medium is subpackaged in the Erlenmeyer flask, picked with an inoculation loop Take a small amount of purified endophytic bacteria in a culture bottle, put the fungus on a shaker at 27°C, 150r / min, and shake it for 6-7 days, put the bacteria on a shaker at 37°C, 150r / min, shake it for 2-3 days, the ...

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Abstract

The invention discloses a separation and selection method of antibiotic antitubercular endophyte, which comprises the steps as follows, the preparation of antibiotic antitubercular plant tissue, the surface sterilization and the sterility test, the separation and the purification of the endophyte, the ferment of endogenetic fungi, the ferment of endogenetic bacterium, the process of the zymotic fluid, the process of the thalli, the selection of the effective antibiotic bacterial strains, the selection of the effective antitubercular bacterial strains and the preservation of the strains. The separation and selection method of antibiotic antitubercular endophyte ensures the realization of obtaining the effective antibiotic antitubercular matter from microorganisms, thanks to separation and selection method, a batch of effective bacterial strains are obtained.

Description

technical field [0001] The invention relates to the technical field of plant drug screening, in particular to a method for separating and screening antibacterial and anti-tuberculosis plant endophytes. Background technique [0002] In recent years, due to the scarcity of plant resources, plant endophytes can produce some special physiologically active substances, so domestic and foreign pharmacists have paid extensive attention to the research of endophytes. In the isolation process of plant endophytes, the disinfection methods used mainly include physical disinfection and chemical disinfection. There are several combinations in the selection of chemical disinfectants: 75% alcohol; 75% alcohol + mercury chloride; 75% alcohol + sodium hypochlorite. Because the disinfection is not thorough, so the selection of physical disinfection and alcohol alone as a disinfectant is less. The combination of the latter two is commonly used. Foreign scholars often use the combination of 75...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/04C12N1/02
Inventor 王伯初邓墨渊杜慧娟魏有章唐玉斌
Owner JIANGSU UNIV OF SCI & TECH
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