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Paenibacillus polymyxa antimicrobial agent and preparation method thereof

A polymyx spore-like and bacillus technology, which is used in the field of biological control of plant diseases and can solve problems such as product differences

Inactive Publication Date: 2017-11-24
YANGZHOU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, it has been found in practice that when preparing biological control fungicides, the products obtained by different preparation methods are significantly different. Therefore, the preparation method for this fungicide needs further research.

Method used

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  • Paenibacillus polymyxa antimicrobial agent and preparation method thereof
  • Paenibacillus polymyxa antimicrobial agent and preparation method thereof
  • Paenibacillus polymyxa antimicrobial agent and preparation method thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0017] Embodiment 1: the selection of bacterial strain CGMCC NO.12410 culture medium

[0018] (1) Inoculate strain CGMCC NO.12410 into NB, LB, KB, Landy, PDB, TSB, GSC culture medium with 5% inoculum amount, set 3 repetitions for each treatment, at 28°C, 180r / min Conditioned culture 3d.

[0019] (2) Use a UV-Vis spectrophotometer to detect its ODat600 value to detect its bacterial concentration.

[0020] (3) Take 40 ml of the culture medium after 3 days of cultivation and centrifuge at 12,000 rpm / min, take the supernatant, and keep the cells for future use. The supernatant was filtered with a 0.22 μm bacterial filter. Add 20% of the filtered supernatant to the PDA and mix the inverted plate. The corresponding sterile medium is mixed with the PDA inverted plate in equal proportions as a control. The center of the plate is inoculated with a tomato gray mold cake with a diameter of 9mm. Each group Set up 3 repetitions. When the control plate is overgrown with gray mold, inves...

Embodiment 2

[0023] Embodiment 2: the selection of cultivation time

[0024] (1) Insert bacterial strain CGMCC NO.12410 into 100ml of KL medium (500ml Erlenmeyer flask) with a 5% inoculum amount, cultivate it at 28°C and 180r / min, and take a sample every 12h. Take 2ml and end the sampling after 48h.

[0025] (2) Use a UV-Vis spectrophotometer to detect the ODat600 value of the obtained sample to detect the bacterial concentration.

[0026] (3) Centrifuge 1ml of the sample at 12000rpm / min, take the supernatant, and keep the bacteria for use. The supernatant was filtered with a 0.22 μm bacterial filter.

[0027] (4) Mix the retained cells with 1ml of methanol solution, extract overnight at 4°C, centrifuge at 12000rpm / min, take the supernatant, and filter with a 0.22μm nylon membrane filter.

[0028] (5) Inoculate the tomato gray mold cake with a diameter of 9 mm to the center of the PDA plate, and after cultivating for 1 to 2 days, punch 4 holes with a diameter of 9 mm at a distance of 12...

Embodiment 3

[0030] Embodiment 3: thalline methanol extract

[0031] Add 1% of the methanol extract of bacteria extracted from KL medium to PDA, mix well and invert the plate, and inoculate rice sheath blight, peach brown rot, cucumber wilt, rice blast fungus, and wheat total eclipse respectively. 8 species of plant pathogenic fungi, including Sclerotinia sclerotiorum, Botrytis cinerea, and Fusarium wilt of cucumber, were investigated for colony diameter with methanol as a control. The results showed that the methanol extract had the best inhibitory effects on peach brown rot, tomato cinerea and rape sclerotinia, and the inhibition rates were 97.7%, 84.2%, and 88.2%, respectively. At the same time, it has a certain antibacterial effect on other pathogenic fungi ( image 3 ).

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Abstract

The invention relates to a Paenibacillus polymyxa antimicrobial agent and a preparation method thereof. The antimicrobial agent is prepared by the following steps: Paenibacillus polymyxa CGMCC No.12410 is fermented on a KL medium for 36 hours, and fermented supernatant or a methanol extract product of bacteria is used as the antimicrobial agent. CGMCC No.12410 is cultivated in LB, PDB, Landy, GSC, NB, KL, and TSC under the same conditions, and antimicrobial capability of the fermented supernatant has large difference. In the same medium, the antimicrobial capability of the fermented supernatant is obviously lower than the antimicrobial capability of the methanol extract product of bacteria. An experiment proves that the bacterial strain CGMCC No.12410 is fermented in the KL medium for 36 hours, the methanol extract product of bacteria has wide antimicrobial spectrum, and the methanol extract product of bacteria can inhibit a plurality of pathogenic fungi. At the same time, diseases prevention and inoculation experiments of Botrytis cinerea which are carried out on tomato fruits show that lowest antimicrobial rate of the methanol extract product of bacteria is 88.6%, and antimicrobial effects are good.

Description

technical field [0001] The invention relates to biological control of plant diseases and belongs to the field of biological control research and application in the discipline of plant protection. Background technique [0002] At present, chemical agents are mostly used to prevent and control plant diseases in production. However, the abuse of chemical agents has caused a series of ecological security and food safety problems, so the use of beneficial microorganisms to control the occurrence and development of diseases has more potential than others. [0003] There are many types of biocontrol bacteria that have been reported, among which Paenibacillus polymyxa is non-pathogenic to plants. It can not only produce plant hormones such as auxin and cytokinin to promote plant growth, but also produce polymyxa Polymyxin, Colistin, Paenibacillin, Gavaserin and Saltavidin, Jolipeptin, Gatavalin, Polypeptins, Fusaricidins and other antibacterial substances inhibit the development o...

Claims

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Application Information

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IPC IPC(8): C12N1/20A01N63/00A01P3/00C12R1/12
CPCC12N1/20A01N63/00
Inventor 张清霞何玲玲童蕴慧
Owner YANGZHOU UNIV
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