A cellulosic-degrading biocontrol bacillus and its application and preparation

A technology for degrading cellulose and Bacillus, applied in the field of microorganisms, can solve the problem of inseparable from high-efficiency cellulose-degrading strains, etc., and achieve the effects of good industrial production prospects, rapid degradation, and shortening the decomposition cycle.

Active Publication Date: 2022-05-24
NORTHWEST A & F UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] However, no matter which of the above-mentioned degradation methods is inseparable from the high-efficiency cellulose-degrading strains

Method used

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  • A cellulosic-degrading biocontrol bacillus and its application and preparation
  • A cellulosic-degrading biocontrol bacillus and its application and preparation
  • A cellulosic-degrading biocontrol bacillus and its application and preparation

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] Isolation and Screening of Cellulose Degrading Bacteria

[0022] 1. The source of soil samples

[0023] Fresh soil samples were taken from the straw returning soil of the Caoxinzhuang Experimental Farm, Northwest Agriculture and Forestry University, Yangling District, Shaanxi Province. After air-drying, the weeds in the soil were screened out, and then the weighed soil was put into a tissue culture bottle. Appropriate amount of water for a week of culture until the microorganisms in the soil recover. After the microorganisms in the soil were recovered, wheat and corn stalks were added to tissue culture flasks at 28°C for 50 days of enrichment culture. Soil samples were collected at 1, 14 and 40 days to isolate cellulose-degrading bacteria.

[0024] 2. Culture medium formula

[0025] (1) Enrichment medium: 5 g of cellulose (straw), 5 g of peptone, 5 g of sodium chloride, 0.5 g of dipotassium hydrogen phosphate, 0.5 g of magnesium sulfate, and 1000 ml of distilled water...

Embodiment 2

[0035] Identification of cellulose degrading strains

[0036] After culturing the strain in liquid medium at 140 rpm shaker for 2 days, DNA was extracted and 16s rDNA sequencing was performed. The sequence result is shown in SEQ ID NO: 1;

[0037] BLAST alignment was performed on NCBI to construct a phylogenetic tree. The result is as image 3 As shown, the similarity between this strain and Bacillus tequilensis is 98%, which belongs to the genus Bacillus (Bacillus sp.) and is named Bacillus sp. CCNWX2.

[0038] The strain was deposited in the General Microbiology Center (CGMCC) of the China Microorganism Culture Collection and Management Committee on June 1, 2020, and the deposit number is CGMCC No: 19895. The deposit address is: Institute of Microbiology, Chinese Academy of Sciences, No. 3, No. 1, Beichen West Road, Chaoyang District, Beijing.

Embodiment 3

[0040] Determination of cellulase activity

[0041] 1. Drawing of glucose standard curve

[0042] Add 0mL, 0.2mL, 0.4mL, 0.6mL, 0.8mL, 1mL of 1mg / mL standard glucose solution to 18 15ml tubes in 3 tubes (3 replicates) respectively, and make up to 1.5ml with distilled water, add 1.5mL DNS solution, shake well, place it in a boiling water bath and continue to heat for 10min, take it out and cool it immediately, fill it up to 15mL with distilled water, shake well, measure its absorbance at a wavelength of 540nm, take the glucose content (mg / mL) as the abscissa, the corresponding The absorbance is the ordinate, and the glucose standard curve is drawn as Figure 4 .

[0043] 2. Determination of cellulase activity

[0044]After the crude enzyme solution was centrifuged at 12,000 r / min, 0.5 mL of the supernatant was added to a colorimetric tube with a stopper that was dried to constant weight in advance, and 1 mL of 1% sodium carboxymethyl cellulose solution was added accurately. ...

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Abstract

The invention discloses a strain of cellulose-degrading biocontrol bacillus and its application and preparation. Its preservation number in the General Microorganism Center (CGMCC) of China Microbiological Culture Collection Management Committee is CGMCC No: 19895. The bacterial strain can efficiently and rapidly degrade the cellulose in the straw, and shorten the straw decomposition cycle; the bacterial strain can antagonize pathogenic bacteria, and can prevent crops from being attacked by pathogenic bacteria after being prepared as a bacterial agent.

Description

technical field [0001] The invention relates to the technical field of microorganisms, in particular to a cellulose-degrading biocontrol Bacillus strain and its application and preparation. Background technique [0002] It is reported that the annual output of straw in my country ranks first in the world. Straw is rich in organic matter and nitrogen, phosphorus, potassium, magnesium, calcium and sulfur and other essential nutrients for crop growth. It is of great significance to give full play to and utilize the positive effect of straw returning to the field, which is of great significance for promoting crop growth and efficient utilization of nutrient resources. . At present, most straws are burned or discarded in the open air, which not only wastes resources, but also causes serious pollution to the environment. Studies have shown that long-term straw returning can increase soil organic matter and improve soil physical and chemical properties. And it can reduce the pol...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20A01N63/22A01P3/00C05F11/08C05F17/20C09K17/14C12R1/07C09K101/00
CPCC12N1/20A01N63/22C05F11/08C05F17/20C09K17/14C09K2101/00C12R2001/07C12N1/205Y02W30/40
Inventor 史鹏罗慧张晶晶张志英张建丽韦革宏
Owner NORTHWEST A & F UNIV
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