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Bacillus subtilis CS30 and application thereof

A technology of Bacillus subtilis and Bacillus marine, applied in application, bacteria, fungicides and other directions, can solve problems such as environmental pollution and food safety, and achieve the effect of improving food and food safety.

Inactive Publication Date: 2019-04-05
QINGDAO UNIV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In addition, the excessive use of chemical pesticides is also likely to cause a series of problems, such as environmental pollution and food security problems caused by pesticide residues, which have aroused people's strong concern

Method used

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  • Bacillus subtilis CS30 and application thereof
  • Bacillus subtilis CS30 and application thereof
  • Bacillus subtilis CS30 and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] Bacillus subtilis CS30 was isolated from deep-sea cold seep sediments in my country. The 16S rDNA sequence of the marine bacterium Bacillus subtilis CS30 is shown in SEQ ID NO.1. After the strain was grown on LB solid medium for 2 days, the colonies appeared milky white, the edges of the colonies were irregular in shape, and many irregular folds appeared on the surface.

[0041] The 16S rDNA (SEQ ID NO.1) sequence of the Bacillus subtilis CS30 is:

[0042] CCTTCGGCGGCTGGCTCCTAAAGGTTACCTCACCGACTTCGGGTGTTAAA ACTCTCGTGGTGTGACGGGCGGTGTGTACAAGGCCCGGGAACGTATTCAC CGCGGCATGCTGATCCGCGATTACTAGCGATTCCAGCTTCACGCAGTCGA GTTGCAGACTGCGATCCGAACTGAGAACAGATTTGTGGGATTGGCTTAAC CTCGCGGTTTCGCTGCCCTTTGTTCTGTCCATTGTAGCACGTGTGTAGCC CAGGTCATAAGGGGCATGATGATTTGACGTCATCCCCACCTTCCTCCGGT TTGTCACCGGCAGTCACCTTAGAGTGCCCAACTGAATGCTGGCAACTAAG ATCAAGGGTTGCGCTCGTTGCGGGACTTAACCCAACATCTCACGACACGA GCTGACGACAACCATGCACCACCTGTCACTCTGCCCCCGAAGGGGACGTC CTATCTCTAGGATTGTCAGAGGATGTCAAGACCTGGTAAGGTTCTTCGCG TTGCTTCGAATTAA...

Embodiment 2

[0044] Example 2 Inhibition of Bacillus subtilis CS30 on Magnaporthe grisea and Gibberella zea.

[0045] The rice blast fungus was inoculated on the potato dextrose solid medium (PDA) and cultured at 28°C for 2-5 days, and then the fermentation broth obtained from the fermentation culture of the bacillus Bacillus subtilis CS30 was filtered through a filter membrane and dropped on the On the sterilized small filter paper sheet, the filter paper sheet is air-dried and placed at a place 2cm away from the blast fungus bacterium block. Culture at 28°C for 3-5 days. After the cultivation, it was observed that the growth of rice blast fungus was significantly inhibited on the culture medium on the side where the Bacillus subtilis CS30 fermentation broth was added dropwise, while the culture medium without the addition of the Bacillus subtilis CS30 fermentation broth was significantly inhibited. Magnaporthe grisea grew well on one side and was not inhibited ( figure 1 ). The experi...

Embodiment 3

[0046] Example 3 Fermentation, separation and identification of active substances produced by Bacillus subtilis CS30.

[0047] The bacterial strain in Example 1 was fermented and cultivated for 48 hours at 28°C, 160rpm liquid medium (peptone 10g / L, yeast powder 5g / L, sodium chloride 10g / L, distilled water 1L, pH 7.0), after fermentation, centrifuged at 8000rpm 15 minutes, collect the fermentation supernatant, adjust the pH of the supernatant to 2.0 with hydrochloric acid, and then overnight at 4°C, collect the acidified precipitated product, extract with methanol, and perform rotary vacuum evaporation on the obtained extracted product to obtain subtilis Crude extract of Bacillus subtilis CS30 active substance.

[0048] The crude extract was further separated and purified by silica gel column chromatography and high performance liquid chromatography (HPLC, UV 210nm). Active substance fractions were obtained by HPLC separation, HPLC used Agilent 1260 series, the mobile phase wa...

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Abstract

The invention belongs to the technical field of screening and application of functional microorganisms, and particularly relates to bacillus subtilis CS30 and application thereof. The bacillus subtilis CS30 capable of producing lipopeptide is collected in China general microbiological culture collection center (address: 3#, Yard 1, Beichen West Road, Chaoyang District, Beijing (Institute of Microbiology, Chinese Academy of Sciences)) on October 31, 2018 with a collection number of CGMCC NO.16666. The bacillus subtilis CS30 of the invention is separated in deep sea cold spring sediment, and hasthe characteristic of restraining the growth of plant pathogenic fungi pyricularia grisea and tumor cells.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to the application of Bacillus subtilis CS30 (marine bacillus). Background technique [0002] At present, for agricultural diseases caused by plant pathogenic bacteria, the usual control method is to use a large amount of chemical pesticides. With the overuse of chemical pesticides, strains of resistant pathogenic bacteria are also emerging. In addition, the excessive use of chemical pesticides is likely to cause a series of problems, such as environmental pollution and food security problems caused by pesticide residues, which have attracted people's strong attention. Chemical pesticides are not only toxic to target organisms, but also accumulate in different organisms along the food chain, and some products also have certain toxic effects such as carcinogenic, teratogenic, and mutagenic effects on humans [1]. As a new type of green pesticide, biopesticides have gradually attracted peo...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20A01N63/00A01P3/00C12P21/02A61K35/742A61P35/00C12R1/125
CPCA01N63/00A61K2035/115A61P35/00C12N1/20C12P21/02C12N1/205C12R2001/125
Inventor 吴仕梅孙超岷
Owner QINGDAO UNIV
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