A high-temperature straw-degrading bacterium b-8 and its agent and application

A technology of straw and bacteria, applied in the field of agricultural resources and environment, to achieve the effect of accelerating the return of field to decompose, broad application prospects and promoting sustainable development

Active Publication Date: 2020-09-08
NANJING AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The currently reported high-temperature cellulolytic bacteria are mainly concentrated between 55°C and 65°C, and there are few reports above 70°C

Method used

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  • A high-temperature straw-degrading bacterium b-8 and its agent and application
  • A high-temperature straw-degrading bacterium b-8 and its agent and application
  • A high-temperature straw-degrading bacterium b-8 and its agent and application

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] Isolation and Identification of Straw Degrading Bacteria B-8

[0031] The straw-degrading bacterial strain B-8 was screened from a straw accumulation place in Weifang City, Shandong Province. Apply the serially diluted sample suspension to the CMC-Congo red medium plate by using the gradient dilution coating method, and cultivate and screen the cellulose-degrading bacteria at 75°C. Purify and isolate the strains that produce larger transparent circles and grow faster on the CMC-Congo red medium plate, inoculate them on the rice straw medium with rice straw as the only carbon source, culture at 75°C, and compare the growth of each strain The bacterial strain B-8, which can efficiently degrade rice straw under high temperature conditions, was finally obtained.

[0032] CMC-Congo red medium: sodium carboxymethylcellulose (CMC-Na) 10.0g, (NH 4 ) 2 SO 4 2.0g, KH 2 PO 4 1.0g, MgSO 4 ·7H 2 O 0.5g, Congo red 0.2g, agar 20g, deionized water 1000mL, pH natural, autoclave...

Embodiment 2

[0038] Inoculate the strain B-8 into the NB medium, and cultivate it at a temperature of 75°C and a rotation speed of 170r / m for 8-12 hours to the logarithmic phase; then inoculate the bacteria B-8 cultivated to the logarithmic phase at an inoculation amount of 1%. Inoculate in rice straw liquid culture medium, cultivate for 7 days at a temperature of 75°C and a rotational speed of 170r / m, remove bacteria and impurities by centrifugal filtration, obtain a crude enzyme solution, and determine endonuclease and β-glucosidase in the crude enzyme solution , xylanase, β-xylosidase activity, see Table 1.

[0039] Rice straw liquid medium: rice straw powder 20g, (NH 4 ) 2 SO 4 1.4g, KH 2 PO 4 2g, MgSO 4 ·7H 2 O0.3g, CaCl 2 2H 2 O 0.4 g, tryptone 1 g, FeSO 4 ·7H 2 O 0.0075g, MnSO 4 ·H 2 O 0.0025g, ZnSO 4 0.002g, CoCl 2 0.003g, 1000mL of deionized water, natural pH, autoclaved at 121°C for 20 minutes; wherein, the rice straw powder is obtained by drying the rice straw to...

Embodiment 3

[0044] Using the method of liquid fermentation in a triangular flask to study the degradation effect of bacteria B-8 on rice straw:

[0045] Preparation of bacterial agent: inoculate strain B-8 into NB culture medium, and cultivate at 75°C and 170r / m for 8-12h to logarithmic phase; then culture B-8 to logarithmic phase 1% inoculum amount was inoculated in rice straw liquid culture medium, cultured for 7 days under the conditions of temperature 75° C. and rotation speed 170 r / m, and centrifugal filtration to remove bacteria and impurities to obtain bacterial agent.

[0046] Pretreatment of rice straw: Soak rice straw in 1% NaOH solution for 30 minutes to remove the wax on the surface of the straw, so that cellulolytic bacteria can better contact with the cellulose in the straw, dry at 50°C, and shred to 2-3cm long.

[0047] Liquid enzyme production medium: (NH 4 ) 2 SO 4 1.4g, KH 2 PO 4 2g, MgSO 4 ·7H 2 O 0.3g, CaCl 2 2H 2 O0.4g, tryptone 1g, FeSO 4 ·7H 2 O 0.0075g...

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Abstract

The invention discloses a high-temperature straw-degrading bacterium B-8, which is classified as Geobacillus Geobacillus sp. , was deposited on March 22, 2019 in the General Microbiology Center of China Committee for the Collection of Microbial Cultures, with the preservation number CGMCC NO.17301. The bacterium B-8 of the present invention can produce degradation circles on the CMC-Congo red medium, and can grow on the medium with crop stalks as the only carbon source. Bacteria B‑8 can produce enzymes related to the degradation of cellulose and hemicellulose, transform the macromolecular substances in crop straws into small molecular substances that can be absorbed and utilized by plants, and realize the resource utilization of agricultural waste under high temperature conditions. Experiments showed that after rice straw was inoculated with B-8 inoculum, it was fermented in liquid at 75°C and 170 r / m for 7 days, and the degradation rate of rice straw was 17.67%. The invention also discloses the application of the bacterium B-8 in degrading crop stalks.

Description

technical field [0001] The invention belongs to the field of agricultural resources and environment, in particular to the field of resource utilization of solid waste, and relates to a strain of high-temperature straw-degrading bacteria B-8 and its bacterial agent, which are used to degrade waste crop straw to produce organic Fertilizers to realize the recycling of agricultural waste. Background technique [0002] my country is a large agricultural country, which generates a large amount of agricultural wastes every year, such as wheat, rice, corn and other crop straws. On the one hand, these straws contain a large amount of nutrients necessary for plant growth, but they mainly exist in the form of cellulose, which cannot be directly used by plants due to their large molecular weight. On the other hand, in recent decades, the application of chemical fertilizers has seriously damaged the health of the soil environment and reduced the diversity of beneficial microorganisms in...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20C05F11/00C05F17/20C12R1/01
CPCC05F11/00C12N1/20C12N1/205C12R2001/01Y02W30/40
Inventor 徐阳春暴彦灼万金鑫陈培杰韦中沈其荣
Owner NANJING AGRICULTURAL UNIVERSITY
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