Preparation and Application of a Strain of Methylotrophic Bacillus and Its Bacterial Agent

A methylotrophic, bacillus technology, applied in the field of agricultural microorganisms, can solve the problems of lack of biological control preparations and methods, enhanced resistance of pathogenic bacteria, soil air pollution, etc. simple craftsmanship

Active Publication Date: 2021-03-30
SHANDONG AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Although chemical control has many advantages such as broad antibacterial spectrum, low cost, convenient use, high efficiency and stability, etc., the extensive use of chemical fungicides brings soil and air pollution, ecological balance damage, food safety threats, and pathogenic bacteria resistance to pesticides. Problems such as plant enhancement are becoming more and more serious. Therefore, people pay more and more attention to the selection of biological control methods that are highly efficient, non-toxic, non-polluting, and not easy to produce drug resistance for the prevention and treatment of plant diseases.
However, the biological control preparations and methods for bacterial diseases of anthurium are very lacking

Method used

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  • Preparation and Application of a Strain of Methylotrophic Bacillus and Its Bacterial Agent
  • Preparation and Application of a Strain of Methylotrophic Bacillus and Its Bacterial Agent
  • Preparation and Application of a Strain of Methylotrophic Bacillus and Its Bacterial Agent

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] 1. Isolation of HZ-9 strain

[0043]Under aseptic conditions, take 5g of the Anthurium rhizosphere soil sample collected, dissolve it in 45mL of sterile water, shake it at 28°C and 220r / min for 1h, absorb 0.5mL and mix it with 4.5mL of sterile water to obtain 10 -1 diluent, and then take 0.5mL 10 -1 The diluted solution is mixed with 4.5mL sterile water to obtain 10 -2 Dilutions, indistinctly, to prepare 10 -3 、10 -4 、10 -5 、10 -6 Wait for a series of dilutions. will be 10 -4 、10 -5 、10 -6 Use a pipette to absorb 100 μL of the diluted solution and add it to the center of the PDA solid medium plate, spread the diluted solution evenly on each PDA solid medium plate with a spreader, and incubate in a biochemical incubator at 37°C for 1-3 days. Use an inoculation loop to pick a single colony of each strain on the plate, streak it on the LB solid medium plate to form a pure culture, and save it.

[0044] Several strains of pure cultures obtained above were subjecte...

Embodiment 2

[0056] Determination of Bacterial Inhibition Spectrum of HZ-9 Strain

[0057] Use an inoculation loop to pick a ring of purified bacteria, and point it at the two ends of the PDA plate at a distance of 3cm. Use the laboratory-preserved grape anthracnose, Fusarium oxysporum, poplar rot, Rhizoctonia solani, Botrytis cinerea Five kinds of pathogenic fungi were used as targets, and the bacteria plates with a diameter of 5mm were cut out with a puncher and inoculated in the center of the PDA plate, and cultured at 28°C for 3-5 days. The test results showed that the HZ-9 strain was effective against grape anthracnose, Fusarium Poplar rot fungus, Rhizoctonia solani and Botrytis cinerea have good antagonistic effects, and the test results are shown in Table 2, attached figure 2 .

[0058] Table 2 Antagonistic effect of HZ-9 strain on various plant pathogenic fungi

[0059]

[0060] Note: "+" indicates that the width of the antibacterial zone is <2mm, "++" indicates that the widt...

Embodiment 3

[0063] 1. Determination of the colonization ability of HZ-9 strain in the rhizosphere of Anthurium and the surface of leaves and petioles

[0064] (1) Double antibiotic screening and stability detection of HZ-9 strain

[0065] The HZ-9 strain was inoculated into LB liquid medium, and cultured with shaking at 30°C and 200r / min for 24h as the seed solution. The seed solution was inoculated into LB liquid medium containing 5 μg / mL rifampicin at an inoculation amount of 2% (volume percentage), and cultured in a shaker flask at 30°C and 200 r / min for 24 hours; the culture solution was inoculated at 2% (volume percentage ) was inoculated into LB liquid medium containing 10 μg / mL rifampicin, and cultured on a shaking table for 24 hours; mL, 80 µg / mL, 150 µg / mL until it increased to 300 µg / mL, and the rifampicin-resistant strains were cultivated.

[0066] The obtained rifampicin-resistant strains were inoculated into LB liquid medium containing 300 μg / mL rifampicin and 5 μg / mL spect...

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Abstract

The invention provides the preparation and application of a methylotrophic bacillus strain and bacterial agent thereof, belonging to the technical field of agricultural microorganisms. The methylotrophic Bacillus HZ-9 of the present invention was deposited in the General Microbiology Center of China Committee for Microbial Culture Collection on April 13, 2017, with the preservation number CGMCC No.14022. The bacterial strain of the present invention has a wider antibacterial spectrum, and has a certain antimicrobial effect on pathogenic bacteria causing anthurium bacterial leaf spot and grape anthracnose, Fusarium oxysporum, poplar rot, Rhizoctonia solani, Botrytis cinerea, etc. Strong inhibitory effect, especially for the pathogenic bacteria that cause bacterial leaf spot of Anthurium, and can stably colonize the rhizosphere, leaves and petiole surfaces of Anthurium; the bacterial agent prepared by using this strain can be applied to anthurium Prevention and control of palm bacterial leaf spot and other diseases.

Description

technical field [0001] The invention provides the preparation and application of a methylotrophic bacillus strain and bacterial agent thereof, belonging to the technical field of agricultural microorganisms. Background technique [0002] Anthurium, also known as anthurium, lucky flower, and anthurium, is a plant of the genus Anthurium in the family Araceae. Its origin is the tropical rainforest area of ​​South America, and it is now widely cultivated in Asia, Europe, Africa and other places. This crop is a perennial evergreen herbaceous flower with peculiar flower shape, bright and rich colors, and a wide range of uses. It is one of the most popular and popular precious flowers in the world. In trade, the sales volume is second only to orchids. [0003] In recent years, the bacterial diseases of Anthurium anthurium have shown an obvious growth trend in the cultivation process of Anthurium, mainly including bacterial leaf spot, bacterial leaf blight, and bacterial wilt. In ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20A01N63/22A01P1/00C12R1/07
CPCA01N63/10C12N1/20C12N1/205C12R2001/07
Inventor 刘训理刘朝阳杨涛王晓辉纪超安雪王爽隋君康王昌乾李倩
Owner SHANDONG AGRICULTURAL UNIVERSITY
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