Bunge pricklyash leaf endophytic bacillus safensis, method for screening and purifying bacillus safensis as well as application thereof

A technology of bacillus and purification method, which is applied in the field of endophytic Bacillus sarfancie and its screening and purification, which can solve the problem of no endophytic Bacillus sarfanciii in the absence of antioxidant properties of endophytic bacterium metabolites and no endophytic Bacillus sarvensis Patent and non-patent literature reports and other issues, to achieve the effect of good antioxidant activity, convenient operation, and simple screening method

Inactive Publication Date: 2015-04-22
SICHUAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] At present, there are no patent and non-patent literature reports on the isolation of endophytic Bacillus sarfenci from Zanthoxylum bun

Method used

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  • Bunge pricklyash leaf endophytic bacillus safensis, method for screening and purifying bacillus safensis as well as application thereof
  • Bunge pricklyash leaf endophytic bacillus safensis, method for screening and purifying bacillus safensis as well as application thereof
  • Bunge pricklyash leaf endophytic bacillus safensis, method for screening and purifying bacillus safensis as well as application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0039] (1) cleaning and disinfection

[0040] In the aseptic operating table, rinse with sterile water three times, soak the fresh red pepper leaves from Hanyuan with 0.5% sodium hypochlorite for 8 minutes, disinfect with 75% ethanol for 10 minutes, rinse with sterile water three times, and keep the water for the last washing as control.

[0041] (2) Enrichment culture

[0042] The surface-sterilized Zanthoxylum bungeanum leaves were ground and inoculated on a nutrient agar medium plate, and cultured in a constant temperature incubator at a temperature of 28° C. for 5 days.

[0043] (3) flat plate coating

[0044] Pick bacterial colonies from the colonies cultured on the above-mentioned nutrient agar solid medium, serially dilute them in sterile saline, spread them on a plate, place them in a constant temperature incubator at 28°C for 7 days, and then pick bacterial colonies on the nutrient agar Streak the plate on the solid medium and place it in a constant temperature inc...

Embodiment 2

[0052] (1) Cleaning and disinfection:

[0053] In the aseptic operating table, rinse with sterile water three times, soak the fresh red pepper leaves from Hanyuan with 1% sodium hypochlorite for 5 minutes, disinfect with 75% ethanol for 8 minutes, rinse with sterile water three times, and keep the water for the last washing as a control .

[0054] (2) Enrichment culture: the surface-sterilized Zanthoxylum bungeanum leaves were ground and inoculated on a nutrient agar medium plate, and cultured in a constant temperature incubator at a temperature of 29° C. for 6 days.

[0055] (3) flat plate coating

[0056] Pick bacterial colonies from the colonies cultured on the above-mentioned nutrient agar solid medium, serially dilute them in sterile saline, spread them on a plate, place them in a constant temperature incubator at 29°C for 6 days, and then pick bacterial colonies on the nutrient agar Streak the plate on the solid medium and place it in a constant temperature incubator a...

Embodiment 3

[0063] (1) Cleaning and disinfection:

[0064] In the aseptic operation table, rinse with sterile water three times, soak the fresh red pepper leaves from Hanyuan with 1.5% sodium hypochlorite for 3 minutes, disinfect with 75% ethanol for 5 minutes, rinse with sterile water three times, and keep the water for the last washing as a control .

[0065] (2) Enrichment culture:

[0066] The surface-sterilized Zanthoxylum bungeanum leaves were ground and inoculated on a nutrient agar medium plate, and cultured in a constant temperature incubator at a temperature of 30° C. for 7 days.

[0067] (3) flat plate coating

[0068] Pick bacterial colonies from the colonies cultured on the above nutrient agar solid medium, serially dilute them in sterile normal saline, spread them on a plate, place them in a constant temperature incubator at 30°C for 5 days, and then pick bacterial colonies on the nutrient agar Streak the plate on the solid medium and place it in a constant temperature in...

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Abstract

The invention discloses a method for screening and purifying bunge pricklyash leaf endophytic bacillus safensis GHF4 and an application thereof, the bacillus safensis is preserved in China Center for Type Culture Collection with a preservation number CCTCC NO: M2014535, and is named as GHF4. The method is characterized by comprising the following steps: taking fresh bunge pricklyash leaf, disinfecting and cleaning, performing plate enrichment culture, performing gradient dilution plate coating, primary screening, secondary screening and the like to realize screening and purifying. its fermentation metabolites has strong antioxidant activity, when the concentration of a broth is 3.75mg/mL, clearance to ABTS free radical by endophytic bacillus safensis can reach 99.48%, and IC50 value is 0.80mg/mL. The endophytic bacillus safensis has good application prospect on production aspect of a natural anti-oxidant.

Description

technical field [0001] The invention relates to an endophytic bacillus sarfenci bacillus on prickly ash leaves, a screening and purification method and application thereof. [0002] The bacterial strain obtained by the present invention has been preserved on October 31, 2014 at the China Type Culture Collection Center, the address of the preservation unit: Wuhan University, Wuhan, China, the preservation number: CCTCC M 2014535, and the classification name: Bacillus safensis GHF4 (Bacillus safensis GHF4 ). Background technique [0003] With the development of society, people have new needs for health. Natural antioxidants have attracted more and more attention because of their wide application range and low toxicity and side effects. The extraction of natural antioxidant active substances has been a research hotspot for many scholars. In recent years, plant endophytes have attracted much attention. Many domestic and foreign studies have confirmed that plant endophytes are...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12Q1/04C12R1/07
CPCC12N1/20C12Q1/04C12N1/205C12R2001/07
Inventor 高鸿白津榕钟凯黄毅娜山口五十磨
Owner SICHUAN UNIV
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