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Bacillus subtilis subsp. spizizenii and application thereof in inhibiting cyanobacterial bloom

A technology of Bacillus subtilis and subspecies, applied in the direction of bacteria, biological water/sewage treatment, and microbial-based methods, etc., can solve the problems of differentiation, evolutionary relationship, and four strains that cannot be used in conventional methods, and achieve broad application prospects , low-cost effect

Active Publication Date: 2021-04-09
TIANJIN UNIVERSITY OF SCIENCE AND TECHNOLOGY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] At present, Bacillus subtilis contains four subspecies, namely Bacillus subtilis subsp.Subtilis, Bacillus subtilis subsp.Inaquosorum, Bacillus subtilis subsp.spizizenii and Bacillus subtilis subsp. .Stercoris, scientific research in recent years has found that these four subspecies can be used as an independent strain, but due to the lack of clearly distinguishable phenotypes, the four strains cannot be distinguished in a conventional way, Christopher A.Dunlap.Michael J. Bowman .DanielR.Zeigler and others compared the four strains from the genome and metabolome, and found that there are differences in the metabolome of the four strains, the main difference is: Bacillus subtilis subsp. Subtilis only produces 3,3'-neotrehalosadiamine; While Bacillus subtilis subsp.Inaquosorum produces bacillus subtilis subsp.Inaquosorum produces bacillus subtilis subsp.spizizenii produces bacillus subtilis subsp.spizizenii, bacitracin and 3 ,3'-neotrehalosadiamine; Bacillus subtilis subsp.Stercoris can produce Fengyuan element and unknown PKS / NRPS structure cluster
In addition, in the comparison of genome differences, the three found that the four types of Bacillus subtilis are monophyletic branches, that is, they have a common ancestor, but the evolutionary relationship between them is currently far away

Method used

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  • Bacillus subtilis subsp. spizizenii and application thereof in inhibiting cyanobacterial bloom
  • Bacillus subtilis subsp. spizizenii and application thereof in inhibiting cyanobacterial bloom
  • Bacillus subtilis subsp. spizizenii and application thereof in inhibiting cyanobacterial bloom

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0054] Example 1 Isolation and identification of Bacillus subtilis subspecies TK812 with algae inhibitory activity

[0055] The Bacillus subtilis subsp.spizizenii TK812 with algae-inhibiting activity in the present invention is isolated from the excrement of wild earthworms collected on the campus of Tianjin University of Science and Technology.

[0056] The specific method is:

[0057] 1. Sample Processing

[0058] Take a captured wild earthworm, wash its surface with sterile normal saline, place it in a test tube, take out the earthworm after it produces excrement, add an appropriate amount of sterile normal saline into the test tube, shake it well and put it in the nutrient broth Dilution coating was performed on the medium, and the coated plate was placed in a 30°C incubator for 48 hours, and a single colony was picked for shake flask culture. The culture condition is 220rpm, 37°C, and after 24 hours of culture, the algae-inhibiting activity microorganisms are screened. ...

Embodiment 2

[0080] Example 2 Inhibition experiment of Bacillus subtilis subspecies TK812 on Anabaena blooms

[0081] Take 6 bottles of healthy Anabaena culture solution (specification: 100ml / 250ml Erlenmeyer flask) in the logarithmic growth phase, numbered 1-6, of which No. 1-3 is used as the blank control group, and the blank nutritional meat is added according to the inoculum amount of 5%. Soup culture medium, No. 4-6 was used as the experimental group, and the enrichment culture solution of bacterial strain TK812 was added according to the same ratio (the enrichment solution of bacterial strain TK812: inoculated in the nutrient broth medium and carried out enrichment culture under the condition of 37°C shaker and 220rpm , cultured until the number of TK812 viable bacteria per milliliter of enrichment solution was 10 8 1), the Anabaena culture solution inoculated with the enriched culture solution of strain TK812 was placed at 28°C, 100rpm, 12h:12h light-dark cycle, and cultured under t...

Embodiment 3

[0088] Example 3 Inhibitory test of Bacillus subtilis subspecies TK812 to Microcystis aeruginosa

[0089] Take six bottles of Microcystis aeruginosa in the logarithmic phase of growth, numbered 1-6, and divide them into two groups on average. The first group is added with 5% nutrient broth blank medium, and the second group is enriched with 5% strain TK812 Mix the culture solution and place it in an incubator at 28°C for 24 hours with a continuous 4000lx light. Samples were taken every 24 hours to measure the absorbance at 730nm. According to the OD 730 Changes to judge the growth of Microcystis aeruginosa, OD 730 The measurement situation is as follows:

[0090] Table 4 strain TK812 to Microcystis aeruginosa OD 730 Impact

[0091]

[0092] It can be seen that the difference from Anabaena is that after adding the blank broth medium, the growth rate of Microcystis aeruginosa in the experimental group is slightly lower than that of the blank control group, and the number o...

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Abstract

The invention belongs to the technical field of biology, and mainly relates to a Bacillus subtilis subsp. spizizenii strain with algal inhibition activity and application thereof in inhibiting and preventing cyanobacterial bloom in shrimp and crab culture ponds. The strain is specifically Bacillus subtilis subsp. spizizenii TK812, and is preserved in China General Microbiological Culture Collection Center on November 3<rd>, 2020, and the preservation number is CGMCC NO. 20996. The Bacillus subtilis subsp. spizizenii is adopted for the first time to prevent and treat cyanobacterial bloom, and the used Bacillus subtilis subsp. spizizenii is separated from the nature, and has no gene modification and no secondary pollution threat; and the microbial inoculum provided by the invention does not contain preservative or chemical additive, has no chemical pollution threat, and has low cost and wide application prospects.

Description

Technical field: [0001] The invention belongs to the field of biological technology, and mainly relates to a strain of Bacillus subtilis subsp. spizizenii with algae-inhibiting activity and its application in inhibiting and preventing cyanobacteria blooms in shrimp and crab breeding ponds. Background technique: [0002] Cyanobacteria, also known as cyanobacteria, are prokaryotic microorganisms that are widely distributed in surface water and can perform photosynthesis. Cyanobacteria bloom refers to the phenomenon that cyanobacteria proliferate abnormally and rapidly in a short period of time due to eutrophication of the water body. Since most cyanobacteria can produce algae toxins that are harmful to aquatic organisms and humans, cyanobacteria blooms are also classified as "harmful algal blooms". Hua (Harmful Algal Bloom, HAB)". In China, the current research on cyanobacteria blooms is mainly concentrated in freshwater waters. Since the 1980s, with the rapid development of ...

Claims

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Application Information

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IPC IPC(8): C12N1/20C02F3/34C12R1/125
CPCC12N1/20C02F3/34
Inventor 王海宽郝忠超张会图王伟
Owner TIANJIN UNIVERSITY OF SCIENCE AND TECHNOLOGY
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