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Bacillus methylotrophicu and 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: 2019-05-31
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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  • Bacillus methylotrophicu and application
  • Bacillus methylotrophicu and application
  • Bacillus methylotrophicu and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] Example 1: Isolation and identification of bacteria

[0022] 1.1 Isolation of bacteria

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

[0024] (2) After diluting the bacterial suspension with sterile water gradient, take 100uL from the bacterial suspension dilution of each concentration and spread it on the LB medium plate, place it at 30°C for 48 hours of constant temperature cultivation, and then pick the plate on the plate. Single colonies with different shapes, sizes and colors were streaked and purified on LB plates and numbered.

[0025] (3) after taking the purified strains of different numbers and culturing them in LB liquid medium at 30 ° C and 180 rpm / min for 24 hours, respectively taking 2 mL of fermentation broth and mixing them into 15 mL of PDA medium, pouring the plate, and after solidification, inoculate the center...

Embodiment 2

[0030] Example 2: Antagonistic effect of methylotrophic Bacillus KNK-1 on Fusarium asiatica (co-culture method)

[0031] (1) After culturing the purified strains with different numbers in LB liquid medium at 30°C and 180rpm / min for 24h, respectively take 2mL of fermentation broth and mix them into 15mL of PDA medium, pour the plate, solidify, and set aside for later use.

[0032] (2) Use a hole punch 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 sheets, which are for later use.

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

[0034] Tabl...

Embodiment 3

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

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

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

[0041] (3) For the first time, spraying was carried out in the early flowering stage of Ningmai 13 varieties of wheat, and 60L of fermentation broth dilution w...

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Abstract

The invention belongs to the technical field of biological pesticide and relates to a strain of bacillus methylotrophicu with high antagonistic activity on wheat scab caused by fusarium asiaticum andfield application, in particular to a kind of bacillus methylotrophicu and application. The preservation number of bacillus methylotrophicu is CGMCC NO:16842, and the preservation unit is the China General Microbiological Culture Collection Center (CGMCC). According to bacillus methylotrophicu and the application, biocontrol bacteria with high inhibitory activity on fusarium asiaticum are screenedout for the first time, and a good control effect is achieved on wheat scab in field plot experiments.

Description

technical field [0001] The invention belongs to the technical field of biological pesticides, and relates to a methylotrophic bacillus with efficient antagonistic activity against wheat scab caused by Fusarium asiatica and its field application, in particular to a methylotrophic bacillus and its application. Background technique [0002] Bacillus methylotrophicus can not only live in the rhizosphere soil, but also colonize inside plants. The bacteria can inhibit a variety of plant pathogens, 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 an antagonistic fungal effect. 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 in the winter wheat areas in th...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20A01N63/02A01P3/00C12R1/07
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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