Bacillus sp. Mangrove-2008, inoculant and application and method of bacillus sp. Mangrove-2008 for inhibiting aspergillus niger growth

A technology of Bacillus and Aspergillus niger, which is applied in the field of inhibiting the growth of Aspergillus niger and Bacillus Mangrove-2008, can solve the problems of high equipment requirements, food safety threats, and inconvenient use, so as to improve the antibacterial effect and avoid adverse effects , Improve the effect of antibacterial efficiency

Active Publication Date: 2019-08-30
HUNAN AGRICULTURAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

The use of chemical methods will lead to residues of related chemicals, which will pose a certain threat or impact on food safety. The use of physical methods has high requirements for instruments and is not easy to use.

Method used

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  • Bacillus sp. Mangrove-2008, inoculant and application and method of bacillus sp. Mangrove-2008 for inhibiting aspergillus niger growth
  • Bacillus sp. Mangrove-2008, inoculant and application and method of bacillus sp. Mangrove-2008 for inhibiting aspergillus niger growth
  • Bacillus sp. Mangrove-2008, inoculant and application and method of bacillus sp. Mangrove-2008 for inhibiting aspergillus niger growth

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Experimental program
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Effect test

Embodiment 1

[0040] The separation and acquisition of embodiment 1 bacillus Mangrove-2008

[0041] Select Shenzhen Baguang mangrove forest (N22°39′, E114°30′) as the collection point, take a small amount of collected sea mud homogenate, and perform conventional 10x gradient dilution in a sterile test tube to obtain a dilution of 10- 2 -10-7 6 gradient samples. from 10- 5 -10- 7 Take 200 μL sample dilutions from the three concentration gradient samples and spread them on the solid separation medium of LB, NA, potato medium PDA, and beef extract peptone medium ND, and observe the colonies after culturing at 37°C for 2-3 days. According to the difference in edge shape, color, and shape, pick typical colonies from the plate and inoculate them on the corresponding isolation medium using the three-section line method to obtain pure single strains. Finally, the isolated pure culture was inoculated into the corresponding slant medium, and stored in a refrigerator at 4°C for future use.

[0042...

Embodiment 2

[0043] Observation and identification of embodiment 2 bacillus Mangrove-2008

[0044] Inoculate the strain Mangrove-2008 on the LB medium, observe and record the state of the colony, and its colony morphology is as follows: figure 1 As shown, the streak culture results are as follows figure 2 shown. Measure the diameter of the colony, the presence of a sterile film, etc.; and perform spore staining and Gram staining on the Mangrove-2008 strain with chemical dyes, and then observe and record the results under an optical microscope. The colony characteristics are shown in Table 1, and the spore staining is as follows image 3 shown.

[0045] The 16S DNA was extracted by conventional methods and amplified by PCR, and the products were sequenced by Hunan Qingke Biotechnology Co., Ltd. The sequencing results were used to construct a phylogenetic tree with MEGA 6.0, such as Figure 4 shown.

[0046] Combining the physiological, biochemical and microbiological detection result...

Embodiment 3

[0049] The culture condition optimization of embodiment 3 bacillus Mangrove-2008

[0050] 1. Screening of strain basal medium

[0051] Five culture media were selected for screening: LB medium, PDA medium, YEB medium, NA medium and broth agar medium. Pick mature strains, inoculate them in the basic liquid medium, and cultivate them at 200r / min, 37°C for 12 hours to make seed liquids, and take 3% of the seed liquids and inoculate them in different mediums, 200r / min, 37°C Constant temperature and shaking culture for 12 hours, observe and compare the results, the concentration of the bacterial solution is measured by the spectrophotometer OD 600 Determination, such as Figure 5 As shown, the results showed that Bacillus Mangrove-2008 had the best growth effect on NA medium, so NA medium was selected as the basic medium for subsequent research.

[0052] 2. Determination of strain growth curve

[0053] Pick mature strains, inoculate them in the basic liquid medium, culture at 2...

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Abstract

The invention belongs to the technical field of biological control and particularly relates to bacillus sp. Mangrove-2008, an inoculant and an application and a method of the bacillus sp. Mangrove-2008 for inhibiting aspergillus niger growth. A collection number of the bacillus sp. Mangrove-2008 is CCTCC (China Center for Type Culture Collection) M2019355, and the bacillus sp. Mangrove-2008 is collected on May 14, 2019. The bacillus sp. Mangrove-2008 is shorter in growth cycle; required fermentation conditions are relatively simple; the bacillus sp. Mangrove-2008 is used for antagonizing the aspergillus niger growth; potential adverse effects of chemical control on environment and grain safety are avoided; the bacillus sp. Mangrove-2008 and a metabolic product of the bacillus sp. Mangrove-2008 can effectively inhibit the aspergillus niger growth; and the bacillus sp. Mangrove-2008 can be processed and developed as a dosage form of the living inoculant and the metabolic product and hasa very important application value.

Description

technical field [0001] The invention belongs to the technical field of biological control, and in particular relates to bacillus Mangrove-2008, bacterial agent and its application and method in inhibiting the growth of Aspergillus niger. Background technique [0002] Aspergillus niger (Aspergillus niger), belonging to the subphylum Deuteromycotina, Hyphophyceae, Hyphosporaceae, Polygonaceae, is a common species in the Aspergillus genus fungus, which exists in moist soil, air and other environments. Under suitable conditions, Aspergillus niger can grow and multiply in large quantities, resulting in a decline in the nutritional quality of grain, the quality of seeds, and the quality of technology and food. It is more likely to produce toxic metabolites, which seriously affect and endanger people's health. Therefore, how to prevent and control the impact of Aspergillus niger on grain, people have taken different measures from the planting, harvesting, transportation and storag...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20A01N63/00A01N63/02A01P3/00C12R1/07
CPCC12N1/20A01N63/00C12R2001/07C12N1/205A01N63/10
Inventor 黄勇郑添鹏阮羽萱何小力阮颖刘博宇谭成方
Owner HUNAN AGRICULTURAL UNIV
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