Fermentation culture solution for producing extracellular antifungal protein by using Bacillus vallismortis SZ-4 and fermentation method of fermentation culture solution

A technology of Bacillus dead valley and fermentation culture liquid, which is applied in the field of fermentation technology of Bacillus dead valley SZ-4 extracellular antibacterial protein, and can solve the problems of soil microecological imbalance, soil pesticide residues, etc.

Active Publication Date: 2020-11-20
JILIN AGRICULTURAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

At present, the control of ginseng root rot is still dominated by chemical control, and the spread of the disease is controlled through the application of carbendazim and h...

Method used

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  • Fermentation culture solution for producing extracellular antifungal protein by using Bacillus vallismortis SZ-4 and fermentation method of fermentation culture solution
  • Fermentation culture solution for producing extracellular antifungal protein by using Bacillus vallismortis SZ-4 and fermentation method of fermentation culture solution
  • Fermentation culture solution for producing extracellular antifungal protein by using Bacillus vallismortis SZ-4 and fermentation method of fermentation culture solution

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Experimental program
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Embodiment 1

[0068] Bacillus vallismortis SZ-4 is preserved by the China Common Microorganism Culture Collection and Management Center, and Fusarium solani (mart.) App.et Wollenw is co-constructed by the provincial department of Jilin Agricultural University for ecological restoration and ecosystem Manage state key laboratory preservation.

[0069] The activation of Bacillus death valley SZ-4 uses beef extract peptone medium (beef extract 3.0g, peptone 10.0g, NaCl 5.0g, agar 16g, deionized water 1000mL, pH 6.8-7.0); the preparation of bacterial suspension uses beef Extract peptone culture solution (beef extract peptone culture medium without agar); the preparation of fermented seed solution adopts BPGM fermentation solution (beef extract 5.0g, peptone 10.0g, yeast extract 5.0g, glucose 5.0g, NaCl 5.0g, MnSO 4 10.0mg, deionized water 1000mL, pH 6.6-7.0); the activation of Fusarium solani used PDA medium (potato starch 5.0g, glucose 20.0g, agar 22.0g, deionized water 1000mL, pH 7.0-7.2). T...

Embodiment 2

[0110] Preparation of bacterial suspension of Bacillus dead valley SZ-4

[0111]The Bacillus dead valley SZ-4 test tube species was activated in beef extract peptone medium, cultured in an artificial climate box at 37°C for 24 hours, passed down for 3 times, a single colony was picked and inoculated into a 250mL triangular flask containing 50mL beef extract peptone culture medium, 32 ℃, 180 rpm constant temperature shaking culture for 24 hours, the obtained bacterial culture solution was centrifuged twice at 4 °C, 4500 rpm, 20 min, the supernatant was discarded, and the obtained bacteria were prepared by the hemocytometer method with a bacterial content of about for 10 8 The bacterial suspension of CFU / mL is ready for use.

[0112] Calculation of the bacterial content of the bacterial suspension: using the hemocytometer method. Spot 10 μL of bacterial fermentation broth on the hemocytometer, record the bacterial cells in the five medium-sized squares in the middle, upper lef...

Embodiment 3

[0115] Preparation of GR4YF microbial preparation

[0116] GR4YF microbial preparation contains the whole fermentation broth culture of Bacillus dead valley SZ-4, the preparation method is as follows:

[0117] (1) Draw 5 mL of the SZ-4 bacterial suspension of Example 2, inoculate it in a 500 mL Erlenmeyer flask containing 150 mL of BPGM fermentation broth, cultivate it at 35° C., 170 rev / min constant temperature shaking for 12 hours, and then test the SZ-4 bacterial strain every 2 hours The fermentation quality of the fermentation liquid is inspected until the growth of the fermentation liquid is relatively full and spores are about to appear, then the fermented seed liquid is obtained, and the fermentation time is 24 to 26 hours;

[0118] (2) Inoculate the fermented seed liquid with an inoculation amount of 4% in a 500mL Erlenmeyer flask containing 200mL GR4Y fermented liquid, shake culture on a shaker, the fermentation temperature is 32°C, the shaker speed is 190 rpm, and th...

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Abstract

The invention discloses a fermentation culture solution for producing extracellular antifungal protein by using Bacillus vallismortis SZ-4 and a fermentation method of the fermentation culture solution. The fermentation culture solution which contains 1000mL of deionized water contains 7-15 g of glucose, 15-25 g of rapeseed powder, 1.5-4.5 g of NaCl, 1.0-3.0 mg of MnSO4.7H2O, 2.5-7.5 mg of MgSO4.H2O, 1.0-3.0 mg of Ca(HCO3)2 and 2.5-7.5 mg of KH2PO4, wherein the pH value of the fermentation culture solution is 5.5-7. The fermentation culture condition combination of the Bacillus vallismortis SZ-4 for producing antifungal protein substances is optimized, and technical support and theoretical guidance are provided for industrial production of the Bacillus vallismortis SZ-4 and acquirement ofhigh yield of the antifungal protein substances. A GR4YF microbial preparation prepared by the fermentation of the Bacillus vallismortis SZ-4 has a good prevention and treatment effect on ginseng rootrot.

Description

technical field [0001] The invention belongs to the technical field of fermentation technology of Bacillus dead valley SZ-4 extracellular antibacterial protein, and in particular relates to a fermentation culture liquid for producing extracellular protein of Bacillus dead valley SZ-4 and a fermentation method thereof. Background technique [0002] Ginseng (Panax ginseng Meyer) is a perennial root medicinal plant of Araliaceae Panax genus, which is widely planted in China, South Korea, Russia, North Korea and other countries. Northeast China is the main production area of ​​Chinese ginseng, and it is planted in a large area at the foot of Changbai Mountain. Due to the special geographical location, the incidence of ginseng root rot caused by Fusarium solani (mart.) App. et Wollenw is relatively serious. At present, the control of ginseng root rot is still dominated by chemical control, and the spread of the disease is controlled through the application of carbendazim and hym...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12P21/00A01N63/22A01P3/00C12R1/07
CPCC12N1/20C12P21/00
Inventor 孙卓杨利民韩梅杨莉韩忠明程林
Owner JILIN AGRICULTURAL UNIV
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