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Methylotrophic bacillus bmf 04 and its fermentation method and use

A methylotrophic and Bacillus technology, applied in the field of marine bacteria, can solve the problems of pesticide resistance of harmful organisms, environmental pollution, pesticide residues, etc.

Active Publication Date: 2021-05-04
HUAIHAI INST OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] The technical problem to be solved by the present invention is aimed at problems such as pesticide residues, environmental pollution, and pest resistance caused by the current application of chemical pesticides to prevent and control plant diseases. Biological pesticides can make up for the shortcomings of chemical pesticides, but there are few types of biological pesticides on the market. To meet the deficiencies of existing technologies such as production needs, provide a new methylotrophic Bacillus (Bacillus methylotrophicus) BMF 04 with strong antibacterial effect and prospects for the development and application of biological pesticides and fertilizers
[0004] Another technical problem to be solved by the present invention is to provide a low-cost, high-efficiency fermentation method for the aforementioned methylotrophic bacillus (Bacillus methylotrophicus) BMF 04 in view of the current situation that needs to save costs and improve fermentation efficiency in the production process

Method used

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  • Methylotrophic bacillus bmf 04 and its fermentation method and use
  • Methylotrophic bacillus bmf 04 and its fermentation method and use
  • Methylotrophic bacillus bmf 04 and its fermentation method and use

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0082] Embodiment 1, Bacillus methylotrophicus (Bacillus methylotrophicus) BMF 04 bacterial strain antibacterial experiment: 1 material and method

[0083] 1.1 Test material

[0084] 1.1.1 Tested strains and pathogenic bacteria

[0085] Methylotrophic Bacillus BMF 04 strain was isolated from Lianyungang sea area. The tested plant pathogenic bacteria (see Table 8) were preserved in this laboratory.

[0086] 1.1.2 Medium

[0087] PDA medium: 200g potato, 20g glucose, 20g agar, 1000mL water;

[0088] PD medium: 200g potato, 20g glucose, 1000mL water;

[0089] BMF 04 strain fermentation medium: bran 1.0%, peanut powder 1.0%, CaCl 2 1.0%.

[0090] 1.1.3 Activation of BMF04 strain and pathogenic fungi

[0091] The BMF04 strain stored at 4°C was transferred to the PDA slant under sterile conditions, and cultured at 28°C for 1d to activate; the phytopathogenic fungi stored at 4°C were transferred to the PDA slant under sterile conditions , cultured at 28°C for 3 days for acti...

Embodiment 2

[0110] Embodiment 2, methylotrophic bacillus (Bacillus methylotrophicus) BMF 04 produces antibacterial substance fermentation medium and shake flask fermentation condition optimization experiment:

[0111] 1. Purpose of the experiment: BMF 04 fermentation medium for producing antibacterial substances and optimization of shake flask fermentation conditions

[0112] 2 Materials and methods

[0113] 2.1 Materials

[0114] 2.1.1 Tested strains

[0115] Methylotrophic Bacillus (Bacillus methylotrophicus) BMF 04 strain, wheat scab (Fusarium gramminearum), isolated and preserved by our laboratory and provided.

[0116] 2.1.2 Medium

[0117] (1) PDA medium: 200 g of fresh potatoes, 20 g of glucose, 20 g of agar, and 1000 mL of water.

[0118] (2) PD medium: 200 g of fresh potatoes, 20 g of glucose, and 1000 mL of water.

[0119] 2.2 Method

[0120] 2.2.1 Strain activation

[0121] The methylotrophic Bacillus BMF 04 strain stored in a refrigerator at 4°C was transferred to PDA s...

Embodiment 3

[0206] Embodiment 3, methylotrophic bacillus BMF 04 solid-state fermentation medium formula and fermentation condition optimization experiment:

[0207] 1 Experiment purpose

[0208]In this experiment, the total number of bacteria and the spore rate in the fermented product were used as detection indicators, and the effects of different carbon sources, nitrogen sources, inorganic salts and their concentrations on the solid-state fermentation of the BMF 04 strain were studied through response surface tests and single-factor test designs. Medium formula; based on the optimized solid-state fermentation medium, optimize the solid-state fermentation conditions.

[0209] 2 Materials and methods

[0210] 2.1 Test material

[0211] 2.1.1 Tested strains

[0212] Methylotrophic Bacillus BMF 04 was isolated, purified and preserved in Lianyungang sea area by our laboratory.

[0213] 2.1.2 Medium

[0214] (1) The strain activation medium is PDA.

[0215] (2) The seed liquid preparati...

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Abstract

The present invention is a methylotrophic Bacillus (Bacillus methylotrophicus) BMF 04, the preservation number of which is CCTCC NO: 2017375. The invention also discloses the use of Bacillus methylotrophic bacillus BMF 04 for inhibiting plant pathogenic fungi, the use of the aseptic fermentation liquid of the strain for inhibiting plant pathogenic fungi, and the use of the aseptic fermentation liquid of the strain as an effective ingredient to make antibacterial drugs. The invention also discloses a method for preparing a sterile fermentation liquid of Bacillus methylotrophic bacteria BMF 04 and a fermentation culture method for producing antibacterial substances. It also discloses the spore-producing method of Bacillus methylotrophic bacteria BMF 04. Solid state fermentation culture method. Aiming at the common pathogens in crops, the invention screens out new marine bacterial strains with broad-spectrum resistance, and adopts strain morphological observation, physiological and biochemical tests and 16S rDNA sequence analysis to determine the species relationship of excellent resistant strains. At the same time, its antibacterial use is also studied, which provides a direction for the application of the strain.

Description

technical field [0001] The invention relates to a marine bacterium, in particular to a methylotrophic bacillus BMF 04, and also relates to its use and fermentation method. Background technique [0002] Biological control has the advantages of no pollution to the environment, safety to humans and other organisms, long-lasting control effect, and no residue in the product. It can make up for the shortcomings caused by chemical pesticides. It is particularly important to screen more excellent strains that have strong antibacterial effects and promote crop growth. important. The existing microbial strains used for the development of microbial pesticides are mainly derived from terrestrial environments. The development and research of microbial strains from the ocean started relatively late. Marine bacteria grow and reproduce quickly and have rich metabolites, which are important germplasm resources for screening resistant strains. Different scholars have isolated a number of ex...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20A01N63/22A01P3/00A01P21/00C12R1/07
CPCA01N63/00A01N63/30C12N1/205C12R2001/07
Inventor 马桂珍陈新元吴海霞万丹丹李欢曹雪梅暴增海
Owner HUAIHAI INST OF TECH