High-temperature-resistant bacillus licheniformis strain NJAU-ND8 for acceleration of compost maturity and application of strain

A technology of NJAU-ND8 and Bacillus genus, applied in the field of agricultural microorganisms, can solve the problems of unstable compost strains and weak effect of compost promotion

Active Publication Date: 2019-07-30
NANJING AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, in the entire composting industry, there are still some shortcomings such as unstable composting strains and weak promotion effect of composting effect. Therefore, it is necess

Method used

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  • High-temperature-resistant bacillus licheniformis strain NJAU-ND8 for acceleration of compost maturity and application of strain
  • High-temperature-resistant bacillus licheniformis strain NJAU-ND8 for acceleration of compost maturity and application of strain
  • High-temperature-resistant bacillus licheniformis strain NJAU-ND8 for acceleration of compost maturity and application of strain

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] Embodiment 1, isolation and identification of functional strains

[0036] Put the samples in the high temperature phase of composting into a 250ml Erlenmeyer flask with glass beads and 100ml of sterile water, shake at 25°C and 170r / min for 20min to form a soil suspension, and draw 10 -2 、10 -3 、10 -4 、10 -5Dilute 0.1ml of the fertilizer suspension, spread it on the lignocellulose culture plate, repeat each concentration 3 times, culture at 28°C for 5 days, pick and purify the growing colonies on the plate for more than 5 times, and select the colonies that can still grow Transfer to slant culture and store at 4°C for later use.

[0037] Through qualitative and quantitative screening, a bacterial strain named NJAU-ND8 was finally obtained. The colony of the bacterial strain NJAU-ND8 on the LB plate was large, pale white, and the edges were scattered. The bacteria were soft and sticky to a certain extent. provoke. The starch decomposition is positive and the tryptoph...

Embodiment 2

[0047] Embodiment 2, NJAU-ND8 produces the determination of various enzyme activities

[0048] Use LB medium as NJAU-ND8 enzyme production medium: 10g peptone, 5g yeast powder, 10g NaCl, 1000ml deionized water, pH 7.0, sterilized at 121°C for 20min. Use 3ml of sterile water to make the bacterial suspension into bacterial suspension, take 0.5ml and inoculate them in the enzyme-producing medium respectively, and culture on a shaking table at 170r / min at 50°C. After 24 hours, the bacterial solution was taken, and after adding the extract solution for extraction, the cell membrane was broken to form a stock solution by using an ultrasonic breaker (300W, ultrasonic for 3s, with an interval of 7s, for 3min).

[0049] Cellulase activity was determined according to the method of Albrecht et al. (2008). The stock solution was mixed with acetate buffer (50mM, pH 5) at a ratio of 1:5 (W / V), shaken on a horizontal shaker for 1h, and then centrifuged at 10000r / min at 4°C for 10min to obta...

Embodiment 3

[0056] Embodiment 3, the generation of liquid inoculum

[0057] Streak the NJAU-ND8 strains stored in glycerol tubes at -80°C on LB solid medium plates for activation, and cultivate them in a 37°C incubator for one hour. Pick a single colony of NJAU-ND8 and culture it in a 3ml liquid test tube at 37°C with shaking at 170r / min for 10 hours, and the bacterial solution is used as the seed solution. With 1% (v / v) inoculum size, a seed solution was transferred to LB liquid shake flask, and cultivated at 37°C and 170r / min until mid-logarithmic phase (OD 600 = 1.0), the cells were collected by centrifugation at 4°C, washed three times with distilled water, and resuspended in an equal volume of distilled water for later use.

[0058] The preparation method of the LB culture medium used is as follows, taking the preparation of 1L medium as an example: protein building 10g, yeast powder 5g, NaCl 10g agar 20g, constant volume to 1000ml, pH natural, sterilized at 121°C for 20min.

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Abstract

The invention provides a high-temperature-resistant bacillus licheniformis strain NJAU-ND8 for acceleration of compost maturity and application of the strain. The preservation number of the bacillus licheniformis strain NJAU-ND8 is CGMCC No.16737, and the strain is applied for acceleration of compost maturity. An application method includes the following steps of uniformly mixing manure of livestock and poultry with auxiliary materials according to a certain C/N rate, then inoculating the liquid strain, adopting a strip-shaped stack fermentation process after strip-shaped stacks are formed, and performing overturning and throwing for fermentation. Compared with a contrast without the strain, the application has the advantages that by adding the strain, temperature raising for the stacks inthe early stage of composting is facilitated, the stacks are pushed to enter a cooling period in advance, thus the increase of a germination index of a compost product is promoted, and the decline ofthe C/N rate of the stacks into a compost maturity range is accelerated.

Description

technical field [0001] The invention belongs to the field of agricultural microorganisms, and provides a strain of Bacillus sp. NJAU-ND8 for accelerating compost maturity and application thereof. Background technique [0002] my country's livestock and poultry breeding industry has entered a stage of large-scale and vigorous development, and large-scale intensive breeding enterprises have emerged rapidly. This ensures the people's demand for meat, eggs and other products, and also produces a large amount of breeding waste. These If the breeding waste cannot be treated effectively, it will become a potential environmental pollution. However, in the "weight loss and drug reduction" action launched by the Ministry of Agriculture, the fertilization of waste from these farms can also provide a large amount of high-quality organic fertilizers that can replace chemical fertilizers on the premise of ensuring the healthy development of the aquaculture industry. Therefore, improving t...

Claims

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Application Information

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IPC IPC(8): C12N1/20C05F15/00C12R1/07
CPCC05F3/00C12N1/20C12N1/205C12R2001/07C05F5/002C05F11/00C05F11/08
Inventor 沈其荣李荣徐谞刘超何宙阳刘东阳
Owner NANJING AGRICULTURAL UNIVERSITY
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