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Preparation and application of a subspecies of Bacillus amyloliquefaciens plant and its agent

A technology of amylolytic spores and bacillus plants, applied in the field of agricultural microorganisms, can solve the problems of lack of resistant varieties, polluted environment, increased resistance of pathogens, etc., and achieve the effect of facilitating industrial production, simple production process, and promoting crop growth

Active Publication Date: 2020-09-29
SHANDONG AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

my country has a vast territory, with different crop types, planting seasons and planting systems, as well as different pathogen types and infection patterns, making it difficult to control; (2) Lack of resistant varieties
Most of the crop varieties promoted in production are susceptible varieties, with few resistant varieties and difficult breeding; (3) The pathogen is difficult to eradicate
The pathogen can overwinter in the soil or on crop residues, and can survive in the soil for many years. The short-term crop rotation control effect is poor, and long-term crop rotation and fallow are difficult to implement in most areas of my country; (4) It is difficult to identify and diagnose
This kind of disease mainly harms the roots and stem bases of plants, the symptoms are hidden, it is difficult to identify early plant disease, and it is difficult to effectively prevent and control; (5) The research on biological control and ecological regulation mechanism is insufficient, and the development of related products lags behind; (6) Chemical fungicides prevent The effective cycle is short, and the control effect is generally better in the early stage, and the effect is weakened until it disappears in the middle and late stages, and the control effect is not ideal.
[0004] At present, the prevention and control of root rot mainly depends on chemical agents. Long-term use of chemical agents leads to increased resistance of pathogens, poor control effect, and pollutes the environment.

Method used

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  • Preparation and application of a subspecies of Bacillus amyloliquefaciens plant and its agent
  • Preparation and application of a subspecies of Bacillus amyloliquefaciens plant and its agent
  • Preparation and application of a subspecies of Bacillus amyloliquefaciens plant and its agent

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] 1. Isolation of XH-9 strain

[0039] Take soil samples from the wheat rhizosphere of the Hanjiazhai wheat field in Fangcun Town, Tai'an City, Shandong Province. Under sterile conditions, take 10g of the soil samples, dissolve them in 90mL of sterile distilled water, and culture them with shaking at 220r / min for 30min. Take 1mL and mix with 9mL of sterile water. Evenly, get 10 -1 diluent, then take 1mL 10 -1 The diluted solution was mixed with 9mL sterile water to obtain 10 -2 Dilutions, indistinctly, to prepare 10 -3 , 10 -4 ,1 0-5 , 10 -6 , 10 -7 Wait for a series of dilutions. The chosen dilution is 10 -5 、10 -6 、10 -7 Use a pipette to draw 100uL of the soil dilution solution in the center of the PDA medium plate, spread the dilution solution evenly with a spreader, and incubate at a constant temperature of 30-32°C for 1-3 days. Pick a single colony, streak it into a pure culture, and save it. The PDA medium components are: 200g potato, 5.0g beef extract, ...

Embodiment 2

[0075] Determination of Bacterial Inhibition Spectrum of XH-9 Strain

[0076] 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, and use Fusarium oxysporum, Colloid anthracnose, Alternaria, Helminthes solani, Helminthes graminearum Fusarium, Fusarium pseudograminee, P. poplar canker and Corinococcus botrytis are 8 kinds of pathogenic fungi as the targets. Use a puncher to cut out a 5mm-diameter fungus plate and inoculate it in the center of the PDA plate, and culture it at 28°C for 2-3 days , the test results show that the XH-9 strain has a good antagonistic effect on 6 strains of pathogenic fungi such as Fusarium oxysporum, colloid anthracnose, Fusarium graminearum, and Alternaria. The test results are shown in Table 2. XH-9 The antagonistic spectrum of the strains is shown in the appendix figure 2 .

[0077] Table 2 The relative bacteriostatic rate of XH-9 strains to various pathogenic fungi

...

Embodiment 3

[0080] 1. Determination of the colonization ability of the XH-9 strain in the rhizosphere of wheat, pepper and corn

[0081] (1) Double antibiotic screening and stability detection of XH-9 strain

[0082] Scrape the lawn of XH-9 strain cultured on 2 ring plates and inoculate it into rifampicin-free LB liquid medium, culture at 30-32°C, 200-220r / m for 24h, as seed solution. The seed solution was inoculated into the LB liquid medium containing 5 μg / mL rifampicin (Rif) with an inoculation amount of 2% (volume ratio), and the shake flask was cultivated for 24h; Inoculated in LB liquid medium containing 10 μg / mL rifampicin, and cultured on a shaking table for 24 hours; and so on, so that the concentration of rifampicin gradually changed from 5 μg / mL, 10 μg / mL, 20 μg / mL, 40 μg / mL, 80 μg / mL, 150 μg / mL until it increased to 300 μg / mL, the strains resistant to rifampicin were obtained by culture. Inoculate the obtained anti-rifampicin strain into the LB liquid medium containing 300 ...

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Abstract

The present invention relates to the preparation and application of a Bacillus amyloliquefaciens plant subspecies and its bacterial agent; the XH‑9 strain is identified as the Bacillus amyloliquefaciens plant subsp. Species, now preserved in the General Microorganism Center of China Microbiological Culture Collection Management Committee, the preservation number is CGMCC NO.13151; Pathogenic fungi such as bacterium, poplar canker and Alternaria have strong antagonistic effects; can stimulate plant growth by producing indole acetic acid and 1-aminocyclopropane-carboxylic acid (ACC) deaminase; can degrade cellulose; can Stable colonization in the rhizosphere of wheat, corn, pepper and other crops; the bacterial agent prepared by using this strain can be applied to the prevention and control of root rot diseases of wheat, corn, pepper and other crops or other soil-borne fungal diseases, and has dual functions of disease prevention and growth promotion The preparation process of the bacterial agent is simple, the fermentation cycle is short, and the cost is low, which is beneficial to industrial production and transportation.

Description

technical field [0001] The invention provides a plant subspecies of Bacillus amyloliquefaciens and the preparation and application of the bacterial agent thereof, belonging to the technical field of agricultural microorganisms. Background technique [0002] Root rot is a general term for root and stem rot diseases caused by fungi, bacteria and nematodes, which can harm hundreds of host plants such as grain, vegetables, cotton, grass and oil crops. In recent years, with the large-scale implementation of continuous cropping, straw returning and no-tillage, the occurrence of root rot has become more and more serious. [0003] Root rot is a soil-borne disease that mainly harms seedlings, and can also occur at the adult plant stage, making it difficult to control. The reasons include: (1) The pathogen is complex. my country has a vast territory, with different crop types, planting seasons and planting systems, as well as different pathogenic types and infection patterns, making...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20A01N63/22A01P3/00A01P21/00C12R1/07
CPCA01N63/00C12N1/20C12N1/205C12R2001/07
Inventor 刘训理王晓辉晁强王爽安雪纪超隋君康洒荣波
Owner SHANDONG AGRICULTURAL UNIVERSITY
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