Bacillus megatherium and suspending agent for preventing and treating grape downy mildew

A technology of Bacillus megaterium and grape downy mildew, which is applied in the biological field to achieve the effect of improving quality and saving expenses

Active Publication Date: 2015-06-03
BIOTECH CENT OF SHANDONG ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

There is no report on the use of Bacillus megaterium to prevent and treat the disease

Method used

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  • Bacillus megatherium and suspending agent for preventing and treating grape downy mildew
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  • Bacillus megatherium and suspending agent for preventing and treating grape downy mildew

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Experimental program
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Embodiment 1

[0027] Strain isolation and identification

[0028] 1. The present invention relates to a kind of biocontrol bacterial strain BMJBN02 of preventing and treating grape downy mildew, which is isolated from the soil of leek field in Shouguang City, Shandong Province. The strain preservation number is CGMCC NO.10478

[0029] 2. Strain identification

[0030] The strain was identified as Bacillus thuringiensis and named BMJBN02 by using the combination method of physiological and biochemical characteristics determination and 16SrDNA sequence comparison analysis. At the same time, the isolated and purified single colonies were identified by V.P test, bacterial film formation test, lecithinase test, starch hydrolysis test, casein hydrolysis test, escin and cellobiose. Refer to the "Common Bacterial System Identification Manual" edited by Dong Xiuzhu, Cai Miaoying, etc.

Embodiment 2

[0032] Preparation of Fermentation Broth and Suspending Concentrate of Biocontrol Bacteria BMJBN02

[0033] 1 Inoculate the above-mentioned biocontrol bacteria BMJBN02 into liquid LB, shake and culture at 30°C and 220rpm for 12-18 hours, take a sample on the ultra-clean workbench to measure the OD value at 600nm, and adjust to zero with uninoculated liquid LB during the measurement process;

[0034] 2 When the OD value of the culture solution is 0.5-0.8, add the seed solution to the culture tank containing the fermentation medium at a volume ratio of 1:150. The culture temperature is 30°C, the rotation speed is 200r / min, the fermentation time is 36-48h, the pH of the fermentation broth is maintained at 7.0, the ventilation volume is 1:1.5, the tank pressure is 0.05-0.1KPa, the sporulation rate is 88%, and the viable count is 5.33×10 9 CFU mL -1 ;

[0035] Among them: corn flour 2%, soybean meal 1%, yeast powder 0.3%, CaCl20.1%, MgSO4·7H2O 0.05%, K2HPO4·3H2O 0.3%, KH2PO40.2%....

Embodiment 3

[0040] Control of Grape Downy Mildew by Biocontrol Strain BMJBN02 and Its Suspending Concentrate

[0041] 1 Serum plate method to determine the inhibitory effect of biocontrol strain BMJBN02 on the germination of grape downy mildew pathogen sporangia

[0042] (1) Acquisition of pathogenic fungus sporangia: collect fresh susceptible leaves, brush off the mildew layer on the lesion with a brush under running water, 22 ° C, relative humidity 90%, alternately cultivate light and dark for 12 hours, and wait for new white to grow. In the case of mold layer, wash the sporangia of the pathogen with distilled water, make a suspension, and adjust the concentration of the sporangia suspension to 1×10 5 CFU mL -1 , stored at 4°C for later use.

[0043] (2) Germination inhibition test (see table 1) of biocontrol bacterium BMJBN02 to grape downy mildew sporangia: adopt LB fermented liquid (cultivate 1 day) 10 times dilutions and do inhibition sporangial germination assay (serum plate meth...

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Abstract

The invention discloses a bacillus megatherium and a suspending agent for preventing and treating grape downy mildew. A strain BMJBN02 of bacillus megatherium for grape downy mildew is preserved in the China microbial preservation management committee general microbiology centre on January 30th, 2015, with the preservation number of CGMCC NO.10478. The invention also discloses a microbial agent produced by utilizing the strain and a preparation method of the microbial agent. The strain BMJBN02 and the suspending agent are high in specificity of the grape downy mildew, and high in control effect; moreover, the average control effect reaches up to more than70 percent.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a bacillus megaterium used for preventing and treating grape downy mildew and a suspending agent thereof. Background technique [0002] Grape downy mildew is a worldwide disease and one of the most important diseases of grapes. It usually begins to occur from May to June, and enters the peak period from August to September. It mainly damages leaves and seriously affects the photosynthesis of leaves. It often causes leaves to scorch and fall early, affecting fruit quality and yield, resulting in a 30% to 50% yield reduction, and severe cases reaching more than 80%. The pathogen belongs to the flagellate subdivision, and the mycelium spreads between the host cells, and extends into the host cells with a spherical haustorium to absorb nutrients. Grape downy mildew mainly damages the leaves, new shoots, tendrils and other tissues of plants; when the disease is severe, the fruit yield de...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20A01N63/00A01P3/00C12R1/11
CPCA01N25/04A01N63/00C12N1/205C12R2001/11A01N2300/00
Inventor 王贻莲李纪顺扈进冬杨合同谢雪迎
Owner BIOTECH CENT OF SHANDONG ACAD OF SCI
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