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Microbial agent and culture method and application thereof

A culture method and bacterial agent technology, applied in the field of microorganisms, can solve the problems of poor antibacterial effect, failure to control diseases, and difficult removal of diseases, increase resistance and adapt to the environment, low cost, and easy to operate Effect

Inactive Publication Date: 2019-10-18
BEIJING UNIV OF AGRI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] However, the above-mentioned control measures all have problems such as high cost of control, long time consumption, and environmental pollution, and the control and antibacterial effect is not good. It is not easy to remove the diseases lurking under the bark, and it cannot achieve a better effect of disease control.

Method used

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  • Microbial agent and culture method and application thereof
  • Microbial agent and culture method and application thereof
  • Microbial agent and culture method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0050] The fermentation culture method of bacterial agent is carried out according to the following steps:

[0051] 1) Use a sterile white pipette tip to pick a single colony on the beef extract peptone solid medium of strain A (T1) and strain B (T2) and inoculate it in the basic liquid medium, and culture it at 37°C for 24 hours as the seed liquid. When the OD600 reaches 0.6 Inoculate the same volume of seed solution (volume ratio 1:1) into 1L of optimum fermentation liquid medium, culture at 30°C for 42 hours, pH 5.5, liquid volume 80mL / 250mL, inoculum volume 3% , at a concentration of 1×10 9 CFU / mL of complex microbial fermentation broth;

[0052] Basic liquid medium (g / L): 0.5g magnesium sulfate, 1g potassium dihydrogen phosphate, 2g tryptone, 10mg vitamin B1, 20g glucose, sterilized at 121°C for 20min;

[0053] Fermentation broth (g / L): 0.5g magnesium sulfate, 1g potassium dihydrogen phosphate, 15g yeast extract powder, 110mg vitamin B, 20g glucose, sterilized at 121°C ...

experiment example 1

[0055] Screening process for optimal ratio between strains:

[0056] Use the Oxford cup method to conduct antagonism experiments between strains. Put the strains inoculated on the beef extract medium into a triangular flask filled with sterile water to make a fermentation broth, and add the fermentation broth with the same concentration as the background bacteria at 40 ° C. About the corresponding sterile liquid culture medium, shake well and pour into a sterile primary Petri dish. After the medium was cooled and solidified, place the sterile Oxford cup on the medium with tweezers after autoclaving, and then draw the same amount of fermentation broth of other strains into the Oxford cup. Each experiment was repeated 3 times. Water served as a blank control. After culturing in the incubator for 2 days, observe whether there is a bacteriostatic zone around the Oxford cup. If there is no bacteriostatic zone, it means that the strain mixed into the plate has no obvious antagonist...

experiment example 2

[0071] Antibacterial effect test of compound bacterial agent in vitro:

[0072] The concentration of the compound bacterial agent fermentation liquid was made into 1×10 by the culture medium. 7 CFU / mL, 1×10 8 CFU / mL and 1×10 9 CFU / mL three concentration gradients of complex microbial fermentation broth.

[0073] Take fresh fruit without obvious diseases and wash it with sterile water to remove the dust on the surface of the fruit, put the fruit in 1% NaClO solution for disinfection for 2 minutes, and then rinse it with sterile water for 3 times. Alcohol was used to disinfect the surface, rinsed again with sterile water for 3 times, and set aside. Use a sterile puncher to injure the middle part of the peach fruit, put the wounds into containers with a concentration of 1×10 7 CFU / mL, 1×10 8 CFU / mL and 1×10 9 CFU / mL three concentration gradients of complex bacterial agent fermentation broth, using sterile medium as a control, soak for 15 minutes, then move the fruit into a ...

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Abstract

The invention relates to the field of microorganisms, and provides a microbial agent. The microbial agent mainly comprises one or two of the following strains of a) Bacillus methylotrophicus T1 whichis preserved in the China General Microbiological Culture Collection Center and b) Bacillus subtilis T2 which is preserved in the China General Microbiological Culture Collection Center; for Bacillusmethylotrophicus T1, the preservation number is CGMCC No.18029, and the preservation time is June 26, 2019; for Bacillus subtilis T2, the preservation number is CGMCC No.18030, and the preservation time is June 26, 2019. The microbial agent has the excellent effect on the bacteriostasis aspect, and particularly has wide application in the aspects of peach brown rot resistance, peach gummosis resistance and lettuce cultivation.

Description

technical field [0001] The invention relates to the field of microorganisms, in particular to a bacterial agent and its cultivation method and application. Background technique [0002] Peach is originally produced in China and has a history of more than 3,000 years. It is a major fruit in the world. China is the country with the longest history of peach cultivation and the largest peach production country in the world. According to statistics, there are more than 1,500 peach resources in my country, including 6 species of light walnut, mountain peach, common peach, Xinjiang peach, Shaanxi-Gansu mountain peach and Gansu peach. In the past ten years, with the improvement of production technology, the peach industry has also developed rapidly. The planting area has increased year by year, and the output of large peaches has increased accordingly. However, there are still some problems in China's peach industry, such as insufficient peach varieties with medium and short cooling...

Claims

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Application Information

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IPC IPC(8): C12N1/20A01N63/00A01P3/00A01P21/00C12R1/125C12R1/07
CPCA01N63/00C12N1/20
Inventor 刘悦萍常昊天王芳侯旭袁雪
Owner BEIJING UNIV OF AGRI
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