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A kind of methylotrophic bacillus and its application

A methylotrophic, bacillus technology, applied in the application, bacteria, fungicides and other directions, to achieve the effect of strong operability, good application prospects, good properties and stability

Active Publication Date: 2022-06-14
BIOTECH CENT OF SHANDONG ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there is no patent related to the control of wheat head blight caused by Fusarium asiaticus with methylotrophic bacillus

Method used

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  • A kind of methylotrophic bacillus and its application
  • A kind of methylotrophic bacillus and its application
  • A kind of methylotrophic bacillus and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] Embodiment 1: Isolation and identification of bacteria

[0022] 1.1 Isolation of bacteria

[0023] (1) In the ultra-clean workbench, place the newly collected diseased ears of wheat in 100 mL of sterile water, shake and rinse for 30 minutes, and suspend them sufficiently to make a bacterial suspension.

[0024] (2) After gradiently diluting the bacterial suspension with sterile water, take 100uL from each concentration of bacterial suspension dilution and spread it on the LB medium plate, place it at 30°C for 48 hours, and then pick it out on the plate Streak and purify single colonies with different shapes, sizes, and colors on LB plates, and number them.

[0025] (3) Take the purified strains with different numbers and culture them in LB liquid medium at 30°C and 180rpm / min for 24 hours, then take 2mL of fermentation broth and mix them into 15mL of PDA medium, pour the plate, after solidification, inoculate in the center of the plate Fusarium asiaticum was suppresse...

Embodiment 2

[0030] Embodiment 2: Antagonism of methylotrophic bacillus KNK-1 to Fusarium asiatica (co-cultivation method)

[0031] (1) After the purified strains with different numbers were cultured in LB liquid medium at 30°C and 180rpm / min for 24 hours, 2mL of fermentation broth was mixed into 15mL of PDA medium, poured onto the plate, solidified, and set aside.

[0032] (2) Use a hole puncher with a diameter of 2.5 cm to punch holes at the edge of the cultured pathogenic fungus Fusarium asiatica P2 colony to obtain bacterial flakes, and set aside.

[0033] (3) Use a sterile pick to pick the above-mentioned P2 bacterium sheet into the center position of the PDA plate in (1), and set the PDA plate only inoculated with the Fusarium asiatica sheet as the control group at the same time, cultivate it at 30°C for 4-6 days, observe Growth colony speed and colony radius of P2. Three replicates were set up for each, and the results are shown in Table 1.

[0034] Table 1 The inhibitory effect o...

Embodiment 3

[0038] Example 3: Detection of the control effect of KNK-1 fermented liquid on field wheat scab

[0039](1) Inoculate KNK-1 into LB culture medium, and culture at 180rpm / min at 30°C for 24h to obtain seed solution with a concentration of about 10 8 cfu / mL;

[0040] (2) Strain KNK-1 was fermented with 60 liters of mechanically stirred stainless steel fermenters with six consecutive tanks: the fermentation medium was molasses medium (1 L): 100 mL of molasses, 1 g of sodium chloride, and 3 g of urea. The liquid volume is 40L, the inoculum volume is 6%, the temperature is 30°C, the stirring speed is 300rpm / min, the ventilation volume is 3L / min, the fermentation time is 2 days, and the biomass reaches 1.50×10 9 cfu / mL to obtain the fermentation broth; dilute the fermentation broth 400 times for subsequent use;

[0041] (3) Spray for the first time at the early flowering stage of Ningmai 13 variety wheat, and apply 60L of fermentation broth dilution per mu. At the same time, the ...

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Abstract

The invention belongs to the technical field of biopesticides, and relates to a methylotrophic bacillus with high-efficiency antagonistic activity against wheat head blight caused by Fusarium asiaticus and its field application, specifically a methylotrophic bacillus and its application. The methylotrophic Bacillus (Bacillus methylotrophicus) of the present invention has a preservation number: CGMCC NO: 16842, and a depository unit: General Microorganism Center (CGMCC) of China Committee for Culture Collection of Microorganisms. The invention screens for the first time the biocontrol bacteria with high inhibitory activity against Fusarium asiatica, and obtains better control effect on wheat scab in the field test.

Description

technical field [0001] The invention belongs to the technical field of biopesticides, and relates to a methylotrophic bacillus with high-efficiency antagonistic activity against wheat head blight caused by Fusarium asiaticus and its field application, specifically a methylotrophic bacillus and its application. Background technique [0002] Methylotrophic Bacillus (Bacillus methylotrophicus) can not only live in the rhizosphere soil, but also colonize inside the plant. The fungus can inhibit a variety of plant pathogens and prevent a variety of diseases, such as tobacco bacterial wilt, cucumber anthracnose and gray mold, rice bacterial blight; also secrete xylanase, chitinase, protease, antifungal Lipopeptide surfactants, etc., have antagonistic fungal effects. Methylotrophic bacillus has no adverse effects on humans and animals, no pathogenicity, and is harmless to the environment. It is a new type of biocontrol bacteria. [0003] In my country, head blight mainly occurs i...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20A01N63/22A01P3/00C12R1/07
Inventor 张新建周红姿周方园赵晓燕张广志吴晓青谢雪迎范素素王加宁
Owner BIOTECH CENT OF SHANDONG ACAD OF SCI
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