A strain of Brevibacterium halotolerant and its application in the preparation of microbial agents against plant pathogenic bacteria

A technology of Brevibacterium halotolerant and microbial agent, applied in the direction of plant growth regulators, microorganism-based methods, microorganisms, etc., can solve the problems of ecological balance and environmental protection threats

Active Publication Date: 2020-09-15
HENAN NORMAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although these chemical pesticides can control diseases and increase yields in the short term, their residues enter the surrounding environment along with rainwater, animal and plant residues, etc., causing huge pressure and threats to ecological balance and environmental protection.

Method used

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  • A strain of Brevibacterium halotolerant and its application in the preparation of microbial agents against plant pathogenic bacteria
  • A strain of Brevibacterium halotolerant and its application in the preparation of microbial agents against plant pathogenic bacteria
  • A strain of Brevibacterium halotolerant and its application in the preparation of microbial agents against plant pathogenic bacteria

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] Pick up a single colony of Brevibacterium halotolerant strain JH8 with a sterilized pipette tip into a conical flask containing 100 mL of LB liquid medium, place it in a shaker at 37 °C and 180 r / min for 24 h. Inoculate 5wt% inoculum into a 250mL fermentation bottle (100mL of LB liquid medium), place in a shaker at 37°C and 180r / min for shaking culture, take the cultured bacterium for 24h, and place it at 4°C at 11000r / min min centrifugation for 15 min, and the supernatant was filtered with a 0.22 μm microporous membrane to obtain a sterile fermentation broth. Take 10mL of sterile fermentation broth and 90mL of melted PDA medium and mix it evenly to make a plate, insert the cultured mycelium block of pathogenic bacteria with a diameter of 6mm, and place it under the condition of 28°C for 4 days. The control colony of the liquid was used as a control, and the pathogenic bacteria of rehmannia root rot was measured by the cross method Fusarium solani According to the inh...

Embodiment 2

[0024] Inoculate the Brevibacterium halotolerant strain JH8 into 300mL LB medium after activation, put it in a constant temperature shaking shaker, shake and culture it at 37°C and 180rpm for 36h, take 30mL of the fermentation broth into a centrifuge tube, and set the temperature at 11000r / min Centrifuge for 15 minutes, take the supernatant and add solid ammonium sulfate to 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% and 100% saturation respectively, fully dissolve and place for 4 ℃ refrigerator, let the protein fully precipitate, and after 24 hours, take out the precipitated extract, and use a low-temperature high-speed centrifuge to centrifuge at 4 ℃, 11,000rpm for 10 minutes to retain the precipitate. Add 1 / 10 of the original volume of the prepared phosphate buffer solution of pH 7.0 to the precipitate, and redissolve the precipitated protein to obtain the crude extract of antibacterial protein with different ammonium sulfate saturation. Filter the reconstituted protein cru...

Embodiment 3

[0026] Pick up a single colony of Brevibacterium halotolerant strain JH8 with a sterilized pipette tip into a conical flask containing 100 mL of LB liquid medium, place it in a shaker at 37 °C and 180 r / min for 24 h. Inoculate 5wt% inoculum into a 250mL fermentation bottle (100mL of LB liquid medium), place in a shaker at 37°C and 180r / min for shaking culture, take the cultured bacterium for 24h, and place it at 4°C at 11000r / min min centrifugation for 15 min, and the supernatant was filtered with a 0.22 μm microporous membrane to obtain a sterile fermentation broth. Take 10mL of sterile fermentation broth and 90mL of melted PDA medium and mix it evenly to make a plate, insert the cultured mycelium block of pathogenic bacteria with a diameter of 6mm, and place it under the condition of 28°C for 4 days. The control colony of the liquid was used as a control, and the cross method was used to measure the pathogen of macular blight Septoria digitalis Pass According to the inhibi...

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Abstract

The invention discloses a strain of Brevibacterium halotolerance and its application in the preparation of antagonizing plant pathogenic bacteria microbial agents, belonging to the technical field of microbial control of plant diseases. The main points of the technical scheme of the present invention are: Brevibacterium halotolerant Brevibacterium halotolerans , preserved in the General Microorganism Center of China Microbiological Culture Collection Management Committee, and the preservation number is: CGMCC NO.14230. The invention also specifically discloses the separation and purification method of the halotolerant Brevibacterium and its application in the preparation of antagonistic plant pathogenic microbial bacterial agents. The present invention adopts the method of plate separation and purification to obtain a strain of bacteria from the rhizosphere soil of the famous and precious traditional Chinese medicine Rehmannia glutinosa, and is identified as Brevibacterium halotolerance by 16s rDNA sequencing. The biocontrol microbial agent provides a new strain resource.

Description

technical field [0001] The invention belongs to the technical field of microbial prevention and control of plant diseases, and in particular relates to a strain of Brevibacterium halotolerant and its application in the preparation of microbial agents for antagonizing plant pathogenic bacteria. Background technique [0002] In recent years, with the progress of society and the rapid increase of population, the demand for crops in various countries in the world has shown a continuous increase trend. According to the survey, in my country, crop production is reduced by 40 million tons each year due to pests and diseases, accounting for about 8.8% of the total output. In the world, crop production is reduced by about 1 billion tons each year due to pests and diseases, accounting for about 1 / 3 of the total output. In order to control pests and diseases and increase crop production and income, a large amount of pesticides are sprayed on farmland every year. Although these chemica...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20C12N1/02A01N63/20A01P3/00C12R1/13
CPCA01N63/00C12N1/02C12N1/20C12N1/205C12R2001/13
Inventor 杨清香王瑞飞王妍李明军
Owner HENAN NORMAL UNIV
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