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A kind of methylotrophic bacillus and its application in agricultural production

A methylotrophic, bacillus technology, applied in the directions of microorganism-based methods, applications, chemicals for biological control, etc., can solve the problem that the effect of disease control is not stable or durable

Active Publication Date: 2021-08-20
山东康地恩生物科技有限公司 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

For example, many bacteria, actinomycetes and fungi prevent the occurrence and spread of diseases by producing antibiotics or competing for nutrients and living space, but the effect of disease control is also affected by complex environmental conditions and cannot be stable or long-lasting

Method used

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  • A kind of methylotrophic bacillus and its application in agricultural production
  • A kind of methylotrophic bacillus and its application in agricultural production
  • A kind of methylotrophic bacillus and its application in agricultural production

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Example 1 strain screening and identification

[0032] 1. Strain screening:

[0033] (1) Soil samples: collected from the soil in the Minsk region of Belarus.

[0034] (2) The soil dilution was prepared by the gradient dilution method, and then plate coating and streak culture were carried out to separate and obtain multiple strains of soil microorganisms; then the transparent circle method was further used to screen out the opposite Rhizoctonia solani, Fusarium solani and Strawberry anthracnose The three strains with the most significant antibacterial effect on pathogenic bacteria such as bacteria were named VB1263, VB1264, and VB1266, respectively.

[0035] 2. Strain identification:

[0036] Morphological characteristics of VB1263 strain: the colony is round, with a diameter of 3-6mm, and the colony has neat edges, smooth and moist surface; after long-term cultivation, the surface of the colony is rough and flat; Gram-positive bacteria can produce spores; the spores...

Embodiment 2

[0040] Example 2 Methylotrophic bacillus VB1263 antibacterial ability evaluation

[0041] 1. Preparation of bacteria solution

[0042] The methylotrophic Bacillus VB1263 was first activated, and then the activated methylotrophic Bacillus VB1263 was picked and inoculated in LB liquid medium, cultivated at 37°C, 220r / min for 14h, and the amount of viable bacteria obtained was 10 8 -10 9 CFU / ml of bacterial liquid.

[0043] 2. Preparation of pathogenic bacteria

[0044] Fusarium oxysporum, Alternaria oxysporum, Sporina graminearum, Botrytis cinerea, Rhizoctonia solani, Agrobacterium tumefaciens, Xanthomonas campestris, Carrot soft rot pectin bacillus 10 kinds of pathogenic bacteria (provided by the Institute of Plant Protection, Shandong Academy of Agricultural Sciences) were inoculated on PDA medium respectively, cultured at 30°C for 5 days, and then used for later use.

[0045] 3. Plate antibacterial test

[0046] Inoculate a pathogenic bacteria cake with a diameter of 7mm...

Embodiment 3

[0051] Example 3 Evaluation of Nitrogen Fixation Effect of Methylotrophic Bacillus VB1263

[0052] 1. Nitrogenase activity assay method:

[0053] (1) Determination of ethylene content

[0054] Add 4 mL of nitrogen-fixing medium to a screw-top test tube with a volume of 21 mL to make a slant. After inoculating the strain to be tested, culture it in a 28°C incubator. Use the empty slant without inoculation as a negative control. Azotobacter was used as a positive control, and 7 replicates were set up. After 72 hours, replace the rubber stopper, inject acetylene gas to a final concentration of 10%, seal with parafilm, and continue to cultivate for 72 hours. Take 100 μL of reaction gas to measure the amount of ethylene produced in a gas chromatograph.

[0055] Standard curve of ethylene content:

[0056] Take 4, 8, 10, 15, 20, 40, 60, and 80 μL of pure ethylene and inject it into a certain volume of screw-top test tube, cultivate it in a 28°C incubator for 72 hours, and measur...

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Abstract

The invention relates to the technical field of functional microorganism screening and application, and specifically provides a new methylotrophic bacillus ( Bacillus methylotrophicus ), and its application in agricultural production is provided. The methylotrophic bacillus is screened from the soil in the Minsk area of ​​Belarus, and the preservation number is CCCTCC NO: M2019439. It has a significant inhibitory effect on various pathogenic bacteria, can effectively prevent and control common crop diseases, and increase crop yield, with broad application prospects.

Description

technical field [0001] The invention relates to the technical field of functional microorganism screening and application, in particular to a methylotrophic bacillus and its application in agricultural production. Background technique [0002] The relationship between microorganisms and agriculture is very close. In the soil known as the "microbial base camp", microorganisms play a major role in the quality cycle and have an irreplaceable role. They break down plant and animal waste and convert it into humus, which contributes to the formation of good soil structure. Many soil microorganisms can fix nitrogen in the air and transform various organic matter, and continuously provide plants with various nutrients such as carbon, nitrogen, phosphorus, and potassium that can be effectively used. Pathogenic microorganisms of insects and antagonistic microorganisms of plant pathogens also widely exist in nature, and they can be used in the control of plant diseases and insect pest...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20A01N63/22A01P3/00C12R1/07
CPCA01N63/00C12N1/20C12N1/205C12R2001/07
Inventor 周英俊苑伟伟凌红丽吕宾孙文丽梁莉
Owner 山东康地恩生物科技有限公司
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