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Bacillus amyloliquefaciens and application thereof to biocontrol on northern corn leaf blight

A technology for dissolving amyloliquefaciens and maize large leaf spot is applied to Bacillus amyloliquefaciens and its application field in the biocontrol of maize large leaf spot, can solve problems such as insufficient and unsatisfactory antagonistic effect, and achieves the effect of easy colonization

Inactive Publication Date: 2016-02-24
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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  • Bacillus amyloliquefaciens and application thereof to biocontrol on northern corn leaf blight
  • Bacillus amyloliquefaciens and application thereof to biocontrol on northern corn leaf blight
  • Bacillus amyloliquefaciens and application thereof to biocontrol on northern corn leaf blight

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Experimental program
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Effect test

Embodiment 1

[0053] Simple identification of bacillus amyloliquefaciens of the present invention to the inhibitory action of Bacillus maize:

[0054] Bacillus amyloliquefaciens (strain D3) of the present invention and maize leaf spot fungus (provided by the disease group of the Institute of Plant Protection, Chinese Academy of Agricultural Sciences) were simultaneously placed on the PDA plate. The strain D3 was connected to the center of the plate by the dot splicing method, and P. maize was connected to the equidistant side of the strain D3, and three replicates and a blank control were set. Incubate at 28°C and record the width of the inhibition zone. According to the width and growth of the inhibitory zone, we can see the inhibitory intensity to maize leaf spot. see Figure 4 , the Bacillus amyloliquefaciens bacteriostasis bandwidth of the present invention shows that it has a high inhibitory efficiency to the growth of Pseudomonas maize.

Embodiment 2

[0056] Further identification of Bacillus amyloliquefaciens of the present invention on the inhibitory action of Bacillus maize:

[0057] A single colony of purified Bacillus amyloliquefaciens (D3 strain) was inoculated on LB medium, and cultured at 30° C. for 48 hours. Pick an inoculation loop into a 250mL Erlenmeyer flask filled with 100mL LB liquid medium, culture at 30°C with shaking at 200rpm / min for 24h. The bacterial suspension was centrifuged at 12000 rpm, and then filtered through a bacterial filter with a pore size of 0.22 μL. Take 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 the bacteria cake of S. maize with a diameter of 5 mm and culture at 25 °C. Sterile water was used as a control. Each treatment was repeated 3 times. After 7 days of culture, the colony diameter was measured by the cross method. Calculate inhibition rate.

[0058] Inhibition rate=[(diameter of control bacteria-diamet...

Embodiment 3

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

[0062] In LB medium, the Bacillus amyloliquefaciens of the present invention was cultured 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 Tween 80) at a concentration of 10 8 cells / mL, stored at 25°C.

[0063] Other comparative tests of storage temperature:

[0064] Stored at 4°C, 20°C, 25°C, and 30°C for 1 month and 2 months respectively, and tested the cell viability of Bacillus amyloliquefaciens. There was no significant difference in cell viability between 1 month and 2 months, so this condition was 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 biocontrol of maize leaf spot. Background technique [0002] Maize leaf spot is a serious leaf blight disease caused by the infection of Setosphaeria turrcica, which causes extensive necrosis of maize leaves, affects photosynthesis, and reduces maize yield. In recent years, due to the lack of corresponding germplasm resources resistant to large spot disease in production and the rapid variation of physiological races of the large spot disease, the large spot disease of maize has appeared in the spring corn regions of Northeast my country and the summer corn regions of Huanghuai Haihai. A clearly aggravated trend. At present, methods such as planting disease-resistant varieties and chemical control are mainly used for maize leaf spot disease. However, due to the drug resistance of S. maize blight and the seriou...

Claims

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Application Information

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