Low-temperature-resistant efficient crop decomposition bacterial liquid and preparation method and use thereof

A low-temperature-resistant technology for crops, applied in the field of low-temperature-resistant and high-efficiency crop decomposing bacteria solution and its preparation, can solve the problems of limiting the degradation function of cellulose-degrading bacteria, low degradation efficiency, and single strain composition

Inactive Publication Date: 2021-06-04
ZHEJIANG ACADEMY OF AGRICULTURE SCIENCES
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] 1. The functional microorganisms in the bacterial agent are composed of medium-temperature or medium-high temperature bacteria. Under low temperature conditions, the microorganisms grow slowly or even stagnate and cannot effectively perform the degradation function
[0004] 2. The composition of bacterial strains is single or unreasonable. In some microbial products, the composition of bacterial strains is single, consisting only of cellulose-degrading bacteria or 2 to 3 kinds of cellulose-degrading bacteria plus fungi, lacking lignocellulose intermediate metabolite-degrading bacteria, easy The metabolic repression effect caused by product accumulation limits the degradation function of cellulose degrading bacteria
[0005] In order to solve this problem, the existing technology is as shown in the patent titled a low-temperature-resistant cellulose-degradable cellulose composite bacterial agent and its preparation method, application number 201811581735.2. Although the functional bacteria in the bacterial agent of this patent are resistant to low temperature, the composition of the bacteria is simple. Composed only of Flavobacterium johnsonii and Stenotrophomonas maltophilia, the degradation rate of the bacterial agent on straw can reach more than 50% in 45 days at 8-16°C, and the degradation efficiency is low

Method used

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  • Low-temperature-resistant efficient crop decomposition bacterial liquid and preparation method and use thereof
  • Low-temperature-resistant efficient crop decomposition bacterial liquid and preparation method and use thereof
  • Low-temperature-resistant efficient crop decomposition bacterial liquid and preparation method and use thereof

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Experimental program
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Effect test

Embodiment 1

[0047] The invention provides a method for preparing a low-temperature-resistant and high-efficiency crop straw decomposing bacterial liquid, comprising the following steps:

[0048] Step 1, enrichment acclimation

[0049] Take 10 mL of soil bacterial suspension according to aseptic technique and put it into 90 mL PCS enrichment medium, carry out enrichment culture at 30 ℃, 150r / min shaker shaking conditions, and select filter paper strips from which the color changes obviously or collapses. The solution was transferred to the next-generation enrichment medium for subculture, and the temperature gradient subculture was reduced by 5°C in each generation to enrich and acclimatize the strains that efficiently degrade straw by volume fraction of 5% (V / V) inoculum .

[0050] Step 2, screening and separation

[0051] Dilute the enriched culture solution in step 1 by a 10-fold gradient method to obtain 10 -3 ~10 -8 For each dilution, draw 100 μL of the dilution and spread it on a...

Embodiment 2

[0061] A kind of low-temperature-resistant and high-efficiency agricultural crop straw decomposing bacterial liquid described in the present invention selects the following 6 kinds of effective active microorganisms: Pseudomonas ( Pseudomonas sp.), Acinetobacter ( Acinetobacter sp.), Trichoderma harzianum ( Trichoderma harzianum Rifai ), Penicillium crustacea ( Eupenicillium crustaceum ), Pediococcus pentosacea ( Pediococcus pentosaceus ), the number of effective viable bacteria in the bacterial fertilizer ≥ 1×10 9 cfu / mL.

[0062] Screening of low temperature resistant and efficient crop straw degrading bacteria.

[0063] According to aseptic operation, use sterile water to make 2 strains of Pseudomonas, 1 strain of Acinetobacter, 1 strain of Trichoderma harzianum, and 1 strain of Penicillium clatices to make a bacterial suspension / spore liquid with the same concentration. Take 5mL and inoculate it into a Erlenmeyer flask containing 45mL of enzyme-producing medium, a...

Embodiment 3

[0069] According to aseptic operation, use sterile water to make 2 strains of Pseudomonas and 1 strain of Acinetobacter into a bacterial suspension / spore liquid with the same concentration, and inoculate 5 mL each into the medium with filter paper as the substrate. ~30°C range, 150r / min shaker culture for 10 days to take samples, and add an appropriate amount (5~10mL) of sulfuric acid and sodium nitrate mixed solution to remove bacteria and insoluble calcium carbonate, pass through an 80-mesh sieve, and sieve the remaining bottom The samples were washed with light buffer of distilled water, dried in an oven at 80°C to constant weight, weighed, and the weight loss rate was calculated. Three parallel samples were used in each group.

[0070] Measurement results such as Figure 6 shown.

[0071] Such as Figure 6 As shown, at 5°C, the filter paper weight loss rate of the three low-temperature resistant cellulose-degrading bacteria is at the lowest level of about 7%; at the opti...

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Abstract

The invention discloses low-temperature-resistant efficient crop decomposition bacteral liquid and a preparation method thereof. The low-temperature-resistant efficient crop decomposition bacterial liquid contains the following decomposition bacteria: pseudomonas sp. PS3-1, pseudomonas sp. SC6-1 and acinetobacter sp. PS1-1, and the effective viable count is greater than or equal to 1*10 <9> cfu/mL. The low-temperature-resistant efficient crop decomposition bacteral liquid has advantages of containing a sufficient nitrogen source required by growth and metabolism of microorganisms and can effectively improve decomposition efficiency of straws without additionally supplementing the nitrogen source during application. Various metabolic pathways of different strains and complementation on different degrading enzyme systems are fused. Pediococcus pentosaceus can effectively degrade intermediate metabolites of straw lignocellulose, such as low-molecular oligosaccharide, lactic acid and ethyl alcohol, and can relieve a repression effect caused by product accumulation in a carbon metabolism process of straws.

Description

technical field [0001] The invention belongs to the field of low-temperature-resistant liquid bacterial fertilizer, and in particular relates to a low-temperature-resistant and high-efficiency crop decomposing bacterial liquid and its preparation method and application. Background technique [0002] Zhejiang Province is a typical double-crop rotation planting area in East China. The winter in this area is damp and cold, the stubble period is short, and the straw is not easy to decompose in a short period of time. Although there are many existing commercially available microbial straw degrading bacterial agent products, most of them have the following problems. [0003] 1. The functional microorganisms in the bacterial agent are composed of mesophilic or medium-high temperature strains. Under low temperature conditions, the microorganisms grow slowly or even stagnate and cannot effectively perform the degradation function. [0004] 2. The composition of bacterial strains is ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12N1/14C12N1/36C05F17/20C12R1/38C12R1/01C12R1/645C12R1/885
CPCC12N1/20C12N1/14C12N1/36C05F11/00C05F17/20Y02W30/40
Inventor 苏瑶杨艳华贾生强沈阿林
Owner ZHEJIANG ACADEMY OF AGRICULTURE SCIENCES
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