Preparation method and application of Bacillus velezensis capable of efficiently antagonizing fusarium graminearum

A technology of Fusarium graminearum and Bacillus, which is applied in the field of microorganisms, can solve problems such as endangering human and animal health, physical health and environmental hazards, and affecting food quality, so as to inhibit germination, inhibit the growth of Fusarium graminearum, improve Effects on Wheat Quality

Active Publication Date: 2020-08-14
HUBEI UNIV +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Therefore seriously affect the food quality with wheat as raw material, endangering the health of people and animals
[0003] At present, chemical agents are mostly used to prevent and control wheat scab, but chemical agents will cause serious harm to human health and the environment

Method used

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  • Preparation method and application of Bacillus velezensis capable of efficiently antagonizing fusarium graminearum
  • Preparation method and application of Bacillus velezensis capable of efficiently antagonizing fusarium graminearum
  • Preparation method and application of Bacillus velezensis capable of efficiently antagonizing fusarium graminearum

Examples

Experimental program
Comparison scheme
Effect test

Embodiment example 1

[0037] Implementation Case 1 Screening of Fusarium graminearum antagonistic bacteria in wheat grains

[0038]Preparation of Fusarium graminearum spores: Inoculate Fusarium graminearum into a 250 mL bottle containing 100 mL of CMC medium, and culture with shaking at 180 rpm and 25°C for 7 days. Filter the culture with two layers of sterile gauze to remove the mycelium to prepare a macroconidia suspension, collect the spores after centrifugation at 10,000 rpm for 20 min at 4 °C, wash the spores twice with distilled water, and use 0.1% (v / v) Tween 20 solution to adjust the concentration of the suspension to 2 x 10 5 CFU / mL.

[0039] Selection of wheat grains against Fusarium graminearum: the wheat grains from the rice-wheat rotation area in Phosphate Mine Town, Zhongxiang County, Hubei Province were washed with 75% alcohol, soaked in 1% sodium hypochlorite solution for 10 minutes, and washed with sterile water. 2 times. The concentration of spore solution is 2×10 5 CFU / mL F...

Embodiment example 2

[0043] Implementation Case 2 Morphological and molecular biology identification of NF002 strain

[0044] Morphological identification: inoculate the screened antagonistic bacteria NF002 in LB liquid medium for overnight activation, spread the obtained bacterial solution on an LB plate, and culture it upside down at 28°C for 2 days, observe the morphological characteristics of the colony, as shown in figure 2 As shown, when cultured for 24 h, the colonies were white, translucent, smooth and moist, slightly raised, round and neat.

[0045] Molecular biology identification: The screened antagonistic bacterium Pickmononas was inoculated into a PA bottle filled with 5 mL of liquid LB medium, cultured overnight at 28°C to produce a large amount of bacterial liquid, and 16S PCR was performed using the bacterial liquid as a template. The PCR amplification system (50 μL) was 27F (5 ， -AGAGTTTGATCCTGGCTCAG-3 ， ) and 1492R (5 ， -TACGGCTACCTTGTTACGACTT-3 ， ) primers 1 μL each, bacter...

Embodiment example 3

[0047] Implementation Case 3 Preparation of Fermentation Broth and Spray Preparation of Bacillus Velez NF002

[0048] Inoculate the screened antagonistic bacteria NF002 in LB liquid medium for overnight activation, inoculate 5% of the inoculum (5% of the volume of the fermentation medium) in the NF002 fermentation medium, and culture with shaking at 28°C and 180 rpm 48 h. After direct gradient dilution of the fermentation broth, spread it on LB plates to count the total bacteria count, take the fermentation broth in a water bath at 80°C for 10 min, and then spread it in a gradient dilution to count the number of spores. After 48 hours of shaking culture, the total number of bacteria in the NF002 fermentation broth reached 3.72×10 10 CFU / mL, the number of spores reached 6.15×10 9 CFU / mL.

[0049] The above fermentation broth was diluted with 0.1% (v / v) Tween 20 to a final concentration of 1.0 × 10 8 CFU / mL, obtained spray preparation. There is no water-insoluble compone...

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Abstract

The invention discloses a preparation method and application of Bacillus velezensis for efficiently antagonizing fusarium graminearum, and belongs to the technical field of microorganisms. The Bacillus velezensis NF002 disclosed by the invention can be used for efficiently antagonizing fusarium graminearum, can effectively inhibit generation of fusarium graminearum DON toxin at the same time, andcan degrade DON toxin in wheat grains. The Bacillus velezensis NF002 can be used as a biological prevention and treatment material for wheat scab, and has good application prospects in the aspect of development of new biopesticides or new biological prevention and treatment microbial agents.

Description

technical field [0001] The invention relates to the technical field of microbes, in particular to a strain of Bacillus veles that efficiently antagonizes Fusarium graminearum and its preparation method and application. Background technique [0002] Wheat scab is one of the major diseases of wheat, which occurs widely all over the world. Wheat head blight can be infected from wheat seedlings to heading stage, mainly causing seedling rot, stem base rot, stalk rot and ear rot, among which ear rot is the most serious hazard. Wheat scab in most parts of my country is caused by Fusarium graminearum ( Fusarium graminearum )caused. The wheat head blight caused by Fusarium graminearum can not only seriously affect the yield of wheat, but also affect the quality of wheat. Deoxynivalenol ( deoxynivalanol , DON) and Zearalenone (Zearalenon) and other toxins. Among them, DON toxin is chemically stable and does not decompose when heated. DON toxin can cause human and animal poisonin...

Claims

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Application Information

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IPC IPC(8): C12N1/20A01N63/22A01P3/00C12R1/07
CPCC12N1/20A01N63/00C12R2001/07C12N1/205
Inventor 倪红靳忻萌孙孝文王行国李亚东杨立军姚伦广
Owner HUBEI UNIV
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