Bacillus amyloliquefaciens and application thereof in prevention and control of citrus bacterial canker

A technology for dissolving starch spores and citrus canker, applied in the field of biological high, can solve the problems of intolerance to storage, deterioration of quality, fruit disease, etc., and achieve the effect of good bacteriostatic effect.

Inactive Publication Date: 2017-08-18
SOUTH CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The fruit is diseased, the quality is deteriorated, the com

Method used

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  • Bacillus amyloliquefaciens and application thereof in prevention and control of citrus bacterial canker
  • Bacillus amyloliquefaciens and application thereof in prevention and control of citrus bacterial canker
  • Bacillus amyloliquefaciens and application thereof in prevention and control of citrus bacterial canker

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] (1) Preliminary screening of Bacillus amyloliquefaciens: Take 1 g of a citrus orchard soil in Hunan Province and add it to 10 ml sterile water, incubate at 200 rpm for 30 min, then dilute with water, the dilution gradient is 10×, 100× and 1000×, take 100 μl of each gradient citri strain (Xac-jx6, Xanthomonas citri pv. citri strain jx-6 strain, referred to as Xac-jx6, the full-length genome of the strain has been published on NCBI GenBank, the sequence number is NCBI ReferenceSequence: NZ_CP011827 .2, on LB solid medium at https: / / www.ncbi.nlm.nih.gov / nuccore / NZ_CP011827.2,). Preparation of LB solid medium containing Xac: use LB liquid medium to culture Xac-jx6 bacteria, cultivate to OD at 28°C, 200rpm 600 At about 1, then mix at 40-50°C at a ratio of 10 μl of bacterial suspension per ml of LB medium, pour about 10ml of medium into each plate to obtain a plate containing Xac-jx6), and culture at 28°C for 48 hours , Observe whether there is a zone of inhibition, and the ...

Embodiment 2

[0050] (1) Preparation of fermentation supernatant

[0051] Prepare 100ml LB liquid medium, inoculate the biocontrol bacteria (ie F9 bacteria) obtained in Example 1 according to the inoculum amount of 1%, and cultivate for 3 days at 28° C. and 200 rpm. Centrifuge the fermentation broth at 8000rpm for 15min, filter the supernatant with a 0.22μm bacterial filter to obtain the fermentation supernatant, take 10μl and add it to the LB solid medium with Xac-jx6 (pre-punched holes with a diameter of 5mm), overnight at 28°C cultivated, the result is as Figure 4 , showing that the supernatant contained antibacterial substances. Then screen the culture medium for the best production of active substances.

[0052] (2) Screening of the medium producing the best active substance

[0053] Prepare 10ml each of eight kinds of liquid media including Landy, YSB, BPY, NYD, PDB, YPG, YPD and corn flour (see Table 3 for the specific composition), adjust the pH value to about 7.0, and pick out ...

Embodiment 3

[0058] (1) Cultivation of strains:

[0059] Inoculate Xac-jx6 single colony LB liquid medium, culture at 28°C, 200rpm, grow to OD 600 =0.6 or so, centrifuge at 5000rpm for 10 minutes to collect bacteria, suspend with sterile water, adjust OD 600 = about 1.0, the xac bacteria solution is obtained;

[0060] Inoculate F9 single colonies into LB liquid medium and culture at 28°C and 200rpm until OD 600 Around 0.6, centrifuge at 5000rpm for 10 minutes to collect the bacteria, suspend with sterile water, and adjust the OD 600 = about 1.0, the F9 bacterial liquid was obtained.

[0061] (2) Inoculation experiment of F9 strain against Xac-jx6 citrus living body

[0062] In order to test the inhibitory effect of biocontrol bacteria F9 in outdoor living organisms, an inoculation experiment of citrus living organisms was carried out. Inoculation mainly adopts the method of needle-pricking leaves to inoculate the bacterial solution. When inoculating, use a 1ml syringe belt (needle), ...

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Abstract

The invention discloses Bacillus amyloliquefaciens F9 and application thereof in prevention and control of citrus bacterial canker. The strain F9 is preserved in China Center for Type Culture Collection in Wuhan University, China on September 26th, 2016 with the preservation number of CCTCC NO:M 2016519. Plate antagonistic experiments find that the strain F9 has citrus bacterial canker inhibitory activity, and an inhibition zone is 26.5mm; supernatant of fermentation broth of the strain F9 after 4 days has antibacterial activity, and the diameter of the inhibition zone can reach 26.5mm; and an in vitro inoculation experiment finds that the strain F9 can inhibit extension and colonization of xanthomonas citri. Therefore, the Bacillus amyloliquefaciens F9 can be applied to prevention and control of the citrus bacterial canker as a biocontrol bacterium.

Description

technical field [0001] The invention relates to the field of biological technology, in particular to a strain of bacillus amyloliquefaciens and its application in preventing and treating citrus canker. Background technique [0002] With the development of science and technology, chemical pesticides have been developed. Because of its high efficiency and broad spectrum, it has been favored all over the world. However, the abuse of chemical pesticides has caused serious ecological and food safety problems, such as: environmental pollution and pesticide residues, which not only destroy the ecosystem, but also affect the value of food and human health. Biological control, on the other hand, uses natural or modified organisms, genes or gene products to reduce the action of harmful organisms in favor of beneficial organisms such as crops, trees, animals and beneficial insects and microorganisms. The biopesticides developed by using the principle of biological control have broad ...

Claims

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Application Information

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IPC IPC(8): C12N1/20A01N63/02A01P1/00C12R1/07
CPCA01N63/10C12N1/20C12N1/205C12R2001/07
Inventor 徐领会王鑫杨小贝史瑜
Owner SOUTH CHINA AGRI UNIV
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