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Bacillus subtilis with antagonism and capable of degrading cellulose at low temperature and application of bacillus subtilis

A Bacillus subtilis, cellulose-degrading technology, applied in applications, chemicals for biological control, bacteria, etc., can solve problems such as broad-spectrum antagonism and different antagonistic abilities

Active Publication Date: 2020-08-11
HEBEI UNIV OF ENVIRONMENTAL ENG
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Bacillus subtilis has been found to be able to antagonize pathogenic bacteria, but the broad spectrum of antagonism and the ability to antagonize are different

Method used

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  • Bacillus subtilis with antagonism and capable of degrading cellulose at low temperature and application of bacillus subtilis
  • Bacillus subtilis with antagonism and capable of degrading cellulose at low temperature and application of bacillus subtilis
  • Bacillus subtilis with antagonism and capable of degrading cellulose at low temperature and application of bacillus subtilis

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0019] The screening and identification process of Bacillus subtilis involved in the present invention are as follows:

[0020] 1. Screening of strains

[0021] Take the straw from the stacked straw pile, rinse the surface with sterile water, spread the rinsed liquid on the beef extract peptone medium, and cultivate it at 15°C.

[0022] 2. Preliminary screening of cellulose degrading strains

[0023] (1) Congo red staining: Inoculate the strains on the beef extract peptone medium onto the sodium carboxymethylcellulose plate by spot inoculation method, inoculate 3 spots on each plate, culture at 15°C for 3 days, and then inoculate with 1 mg / mL Congo red staining solution was used for staining for 30 minutes, the dyeing solution was discarded, and 1mol / L NaCl solution was added to decolorize for 20 minutes. Observe whether there is a hydrolysis circle near the bacterial colony, if there is, it means that the strain has the ability to degrade cellulose, and record the size of t...

Embodiment 2

[0035] In this embodiment, cellulase activity was measured on GN19, and the process was as follows:

[0036] Inoculate the primary screened strains into the seed culture medium, and cultivate them at 15°C and 150r / min for 12h to obtain the seed liquid. The seed solution was inoculated into 100mL liquid fermentation medium at 2%, and cultivated under the same conditions for 72h. The obtained fermentation liquid was centrifuged at 4°C and 10,000 r / min at high speed for 15 min, and the supernatant was collected to be the crude enzyme liquid.

[0037] (1) Endoglucanase activity: get 4 test tubes, 1 as a control and 3 as a test tube, add 1.5 mL of 1% CMC-Na solution respectively, add 0.5 mL of crude enzyme solution in the test tube, wherein Add 2mL DNS to the control to inactivate the enzyme activity, and react at 50°C for 30min. After the reaction, add 2mL DNS to the 3 test tubes to terminate the reaction, shake well and then boil in water bath for 10min, distill the volume to 2...

Embodiment 3

[0045] In this embodiment, antagonism test is carried out on GN19 bacterial strain, and the process is as follows:

[0046] Inoculate the plant pathogenic bacteria cake in the center of the PDA solid medium, use a toothpick to pick up Bacillus subtilis GN19 at a distance of 2cm from the pathogenic bacteria cake, directly inoculate the pathogenic bacteria cake as a control, and culture at a constant temperature of 28°C for 3 to 7 days, measure the radius of the colony, and calculate the antibacterial effect rate, the results are shown in Table 3, with another Bacillus subtilis DN5 as a comparison.

[0047] Bacterial inhibition rate = [(control colony radius - treatment colony radius) / control colony radius] × 100%

[0048] Table 3 Inhibitory rate of Bacillus subtilis GN19

[0049]

[0050]

[0051] It can be seen from the above table that the GN19 strain has an antagonistic effect on a variety of pathogenic bacteria, especially the bacteriostatic rate of Sclerotinia scler...

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Abstract

The invention discloses bacillus subtilis with antagonism and capable of degrading cellulose at low temperature and application of the bacillus subtilis, and belongs to the technical field of biology.The bacillus subtilis has various antibacterial activities, is particularly used for sclerotinia sclerotiorum and potato scab bacteria, and is preserved in China General Microbiological Culture Collection Center with the preservation address of Institute of Microbiology, Chinese Academy of Sciences, No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing. The bacillus subtilis GN19 is preserved on January 09, 2020, the preservation number is CGMCC No.19320, the 16S rDNA sequence length of the bacillus subtilis GN19 is 1482bp, and the sequence of the bacillus subtilis GN19 is shown as SEQID NO.1.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a bacillus subtilis strain capable of antagonizing low-temperature degradation of cellulose and an application thereof. Background technique [0002] my country is a large agricultural production country. With the substantial increase in grain production, a large amount of agricultural solid waste is also produced, among which the agricultural solid waste with the largest output is straw. Although there are multiple ways of resource utilization of straw, the utilization rate of straw is still very low in most areas. Therefore, the random discarding of a large amount of straw has caused serious resource waste and economic loss to a certain extent. At the same time, when the straw is burned, it will produce a large amount of polluting gas. Once it enters the atmosphere, it will cause serious air pollution. This pollution is also an important source of agricultural non-poin...

Claims

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Application Information

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IPC IPC(8): C12N1/20A01N63/22A01P1/00A01P3/00C12R1/125
CPCA01N63/22C12N1/205C12R2001/125
Inventor 刘佳莉何鑫宋瑜殷博王书功杜洁柏祥
Owner HEBEI UNIV OF ENVIRONMENTAL ENG
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