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A kind of Bacillus amyloliquefaciens and its application in biocontrol of maize leaf spot

A technology of Bacillus amyloliquefaciens and Bacteria maize, applied in the field of Bacillus amyloliquefaciens and its application in the biological control of Bacillus amyloliquefaciens, can solve the problems of insufficient antagonism and not ideal antagonism, and achieve the effect of easy colonization

Inactive Publication Date: 2018-11-20
INST OF PLANT PROTECTION CHINESE ACAD OF AGRI SCI +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] Although some microorganisms have been screened to have biocontrol effects on P. maize, it is far from enough
On the one hand, the diversified use of antagonistic microorganisms in biocontrol is beneficial to prevent pathogens from developing resistance, so the more types of microorganisms that have antagonistic effects on pathogens, the better; It is not ideal enough, and it is necessary to screen microorganisms with stronger colonization ability and antagonistic effect, so that biological control methods can better meet the requirements of preventing and controlling maize leaf spot in agricultural production

Method used

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  • A kind of Bacillus amyloliquefaciens and its application in biocontrol of maize leaf spot
  • A kind of Bacillus amyloliquefaciens and its application in biocontrol of maize leaf spot
  • A kind of Bacillus amyloliquefaciens and its application in biocontrol of maize leaf spot

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0053] The simple identification of the inhibitory effect of the Bacillus amyloliquefaciens of the present invention on the pathogen:

[0054] The Bacillus amyloliquefaciens (strain D3) of the present invention and the pathogen of corn leaf spot (provided by the disease group of the Institute of Plant Protection, Chinese Academy of Agricultural Sciences) are simultaneously connected to the PDA plate. The strain D3 was connected to the center of the plate by the dot connection method, and the E. maize strain was connected to the equidistant side of the strain D3, and 3 replicates and a blank control were set. Incubate at 28°C and record the width of the inhibition zone. According to the width of the inhibition zone and the growth situation, the intensity of the inhibition of corn leaf spot disease can be seen. See Figure 4 The bacteriostatic bandwidth of Bacillus amyloliquefaciens of the present invention indicates that the growth inhibition efficiency of Bacillus amyloliquefaci...

Embodiment 2

[0056] Further identification of the inhibitory effect of the Bacillus amyloliquefaciens of the present invention on the pathogen of corn leaf spot:

[0057] The purified Bacillus amyloliquefaciens (D3 strain) was inoculated with a single colony on LB medium and cultured at 30°C for 48 hours. Pick an inoculation loop and insert it into a 250 mL Erlenmeyer flask containing 100 mL of LB liquid medium, and culture it with shaking at 200 rpm / min at 30°C for 24 hours. The bacterial suspension was centrifuged at 12000 rpm and filtered through a 0.22 μL bacterial filter. Pipet 1 mL of the filtrate and add it to 19 mL of PDA medium at about 45°C, mix well, and after the medium is cooled, insert a 5mm diameter Pseudomonas corniculata bacteria cake and cultivate at 25°C. Sterile water is the control. Each treatment was repeated 3 times. After 7 days of culture, the colony diameter was measured by the cross method. Calculate the inhibition rate.

[0058] Inhibition rate = [(control bacte...

Embodiment 3

[0061] The preparation method of the biocontrol agent (suspension bacterial agent) of the present invention:

[0062] LB medium, culture the Bacillus amyloliquefaciens of the present invention for 3 days at 30°C to produce a large number of spores. The spores were formulated into a suspension with sterile water (containing 0.05% by weight of Tween 80) with a concentration of 10 8 Pieces / mL, store at 25°C.

[0063] Comparison test of other storage temperature:

[0064] Stored at 4°C, 20°C, 25°C, and 30°C for 1 month and 2 months, respectively. The viability of Bacillus amyloliquefaciens was tested. It was found that the vigor of the bacteria was the strongest at 25°C, and there was no decline in the storage period. There is no significant difference between 1 month and 2 months of cell viability, so this condition is selected as the storage condition for spores.

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Abstract

The invention discloses bacillus amyloliquefaciens and application thereof to biocontrol on northern corn leaf blight. 16S rDNA of the bacillus amyloliquefaciens contains a sequence shown as SEQ ID NO: 1. The preferable bacillus amyloliquefaciens is preserved at China General Microbiological Culture Collection Center, and the preservation number is CGMCC No.11488. The provided application means application of the bacillus amyloliquefaciens to inhibit corn leaf blight pathogen. The bacillus amyloliquefaciens provides an easily-colonized novel microbe with strong antagonistic action for biocontrol on corn leaf blight. Through application of bacillus amyloliquefaciens to biocontrol on corn leaf blight, an efficient stable biocontrol method for corn leaf blight is provided, the characteristic is utilized that bacillus amyloliquefaciens can easily colonize at corn leaves and possesses strong antagonistic action on corn leaf blight pathogen, and requirement about control on corn leaf blight in agricultural production is relatively well satisfied.

Description

Technical field [0001] The invention relates to a Bacillus amyloliquefaciens and its application, in particular to a Bacillus amyloliquefaciens and its application in the biocontrol of corn spot disease. Background technique [0002] Corn leaf spot disease is a serious leaf blight disease caused by Setosphaeria turcica infection, which causes large-area necrosis of corn leaves, affects photosynthesis, and reduces corn production. In recent years, due to the lack of corresponding germplasm resources for resistance to large leaf spot disease in production, and the rapid variation of the physiological races of the large leaf spot disease bacteria, the large leaf spot disease appeared in spring maize areas in northeast my country and Huanghuaihai summer corn areas. Obviously aggravate the trend. At present, methods such as planting disease-resistant varieties and chemical control are mainly used for corn leaf spot disease. However, due to the pesticide resistance of the pathogen and...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20A01N63/00A01P3/00C12R1/07
Inventor 刘文德田雪亮
Owner INST OF PLANT PROTECTION CHINESE ACAD OF AGRI SCI
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