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A Strain of Bacillus and Its Application

A technology of Bacillus and Bacillus Veles, applied in the field of microorganisms, can solve the problems of cost and low yield hindering the application, and achieve good colonization effect, high safety, and the effect of reducing plant diseases

Active Publication Date: 2021-11-19
SHANDONG BEE LAN BIOTECHNOLOGY CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the high cost and low yield of producing Surfactin hinder its application. Obtaining high-yielding strains of Surfactin is the most effective way to reduce costs. On the one hand, it can screen high-yielding wild strains from the environment, select and breed excellent strains, and optimize the fermentation process. , look for cheaper raw materials and efficient separation and purification methods to reduce costs

Method used

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  • A Strain of Bacillus and Its Application
  • A Strain of Bacillus and Its Application
  • A Strain of Bacillus and Its Application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0053] Example 1 Screening and identification of strains

[0054] Screening of strains

[0055] 1. Test material

[0056] 1.1 Soil

[0057] The soil was collected from vegetable gardens around Tai'an City, Shandong Province.

[0058] 1.2 Pathogenic bacteria

[0059] Root rot pathogens and gray mold pathogens were provided by Shandong Baolai Lilai Bioengineering Co., Ltd. Strain Resource Collection Center.

[0060] 1.3 Medium

[0061] PDA: 200g potato, 20g glucose, 2% agar, 1L water; LB: 10g peptone, 5g yeast powder, 10g NaCl, 1L water, pH7.0; blood agar plate was purchased from Shanghai Chemagar Biotechnology Co., Ltd.

[0062] 1.4 Standards

[0063] Surfactin standard products were purchased from Sigma Company.

[0064] 2. Test method

[0065] 2.1 Isolation and purification of strains

[0066] Take 10g of the collected soil, put it into 90mL sterile water, shake and mix well, dilute it into a suitable gradient, spread it on an LB plate, and incubate it in a 37°C in...

Embodiment 2

[0127] Example 2 3.5 Optimization of liquid fermentation medium and culture conditions

[0128] 3.5.1 Screening of fermentation medium

[0129] The best fermentation medium for K-19 to produce surfactin was screened by comparing 8 culture media that were favorable for the formation of antimicrobial lipopeptides.

[0130] Screening method: In a 250ml Erlenmeyer flask, use the seed medium to activate and cultivate for 24 hours, then inoculate 15 different fermentation medium with 2% inoculum size, rotate at 170r / min, temperature 37°C, liquid volume 50ml, naturally cultured under pH conditions. Each medium was replicated in triplicate. After culturing for 48 hours, samples were taken to detect the content of surfactin in each bottle of fermentation broth to determine the optimal medium.

[0131] Under the same fermentation conditions, taking the output of LB medium as a control, the yields of surfactin in No. 6 and No. 7 mediums were significantly increased (657.16, 607.21m...

Embodiment 3

[0156] 3.6 Microbial agent biocontrol test

[0157] Bacterial solution preparation method: soybean meal 45g / L; G10 g / L; MgSO 4 0.2g / L; MnSO 4 2.5mg / L; FeSO4 0.8mg / L, L-glutamic acid 3mg / L, the balance is water; рН6.8; temperature 35°С, stirring speed 160r / min, incubation time 42h. Obtain Bacillus velezensis K-19 fermentation broth. The fermented liquid was diluted 200-500 times to select different crops for pot control effect experiment and field experiment.

[0158] 3.6.1 Growth promotion experiments on peanuts

[0159] The test field is selected from a farmer's open-air peanut field in Linyi, Shandong, and the growth is not good, so the growth-promoting experiment is carried out on peanuts using the bacterial agent of the present invention. Two groups were set up in the test, each group had three ridges of peanuts, and the experimental group used 2% bacterial agent of the present invention (diluted 200-500 times), flushed the roots, applied the second time at intervals...

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Abstract

The present invention relates to the technical field of microorganisms, in particular to a strain of Bacillus and its application. The Bacillus is Bacillus velezensis K‑19, which has been preserved in China's Typical Culture on November 2, 2020. Biological Preservation Center (Address: Bayi Road, Wuchang District, Wuhan City, Hubei Province, Wuhan University), and its biological deposit number is CCTCC NO: M 2020672. Under the optimized fermentation conditions of K‑19, the yield of surfactin is not less than 1.4g / L, and the sum of the yields of surfactin and iturinA can reach 1.82g / L; its metabolites are natural antibacterial agents with no toxic and side effects , has basically no hemolytic activity, does not produce drug resistance, and has good plant growth promotion and biocontrol effects. Under field conditions, it is used in the cultivation of ginger and peanuts, and the promotion effect on peanut plants is significantly higher than that of the control group. And it can inhibit the senescence of cucumber plants, reduce disease spots, and prolong the growth period, which proves that it has good colonization effect, can adapt to field environment, can promote plant growth, and reduce plant diseases.

Description

technical field [0001] The invention relates to the technical field of microorganisms, in particular to a bacillus strain and its application. Background technique [0002] The information disclosed in this background section is only intended to increase the understanding of the general background of the present invention, and is not necessarily taken as an acknowledgment or any form of suggestion that the information constitutes the prior art already known to those skilled in the art. [0003] Surfactin is a secondary metabolite produced by Bacillus sp. strains. It is a cyclic lipopeptide composed of a seven-membered peptide and a β-hydroxy fatty acid chain. There are many homologues. According to Carbon chain length can be divided into C 12 —C 16 5 components. Compared with general chemical surfactants, in addition to reducing surface tension, stabilizing emulsion and increasing foam, it also has the advantages of thermal stability, non-toxicity, biodegradability and st...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20A01N63/22A01P1/00A01P3/00A01P21/00C12R1/07
CPCA01N63/22C12N1/20
Inventor 胡著然王丽荣冉新炎韩广泉陶宁齐素敏陈丹丹李圆圆张文娟申小冉董雪雁
Owner SHANDONG BEE LAN BIOTECHNOLOGY CO LTD
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