Bacillus amyloliquefaciens, microbial agent as well as preparation method and application of microbial agent

A technology of amylolytic spores and bacillus, applied in the field of microorganisms, can solve the problems of environmental pollution by chemical agents, unsatisfactory control effects, and drug resistance of pathogenic bacteria, and achieve good control effects, good stability, and high spore production.

Active Publication Date: 2021-03-26
QINGDAO AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] In production, people mainly use chemical pesticides to prevent and control the above diseases, but the long-term use of chemical agents can easily cause pathogenic bacteria to develop drug resi...

Method used

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  • Bacillus amyloliquefaciens, microbial agent as well as preparation method and application of microbial agent
  • Bacillus amyloliquefaciens, microbial agent as well as preparation method and application of microbial agent
  • Bacillus amyloliquefaciens, microbial agent as well as preparation method and application of microbial agent

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Experimental program
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Effect test

Embodiment 1

[0044] The isolation and screening of embodiment 1 bacterial strain F028

[0045] In the tomato planting field in Weifang City, Shandong Province, the surface soil was shoveled off, and about 500g of soil at a depth of 5-10cm was taken, put in a sterilized ziplock bag, and brought back to the laboratory. Strains were isolated by conventional dilution plate method, and antagonistic strains were screened with the pathogen of tobacco red spot disease as indicator bacteria. As a result, a strain with strong antagonistic effect was obtained, which was named F028.

Embodiment 2

[0046] Identification of embodiment 2 bacterial strain F028

[0047] 1. Morphological Identification

[0048] The strains were inoculated on NA plates, incubated at a constant temperature of 30°C for 12-48h, and the characteristics of the colonies were observed. The strains cultured for 12 hours were picked for Gram staining, and the strains cultured for 48 hours were picked for spore staining. The results showed that the colony was colorless and transparent at the initial stage, with a smooth, moist, sticky surface and irregular edges, and in the later stage, it was milky white with obvious bulges and wrinkles on the surface. Bacteria rod-shaped, spore-forming, Gram-positive. Such as figure 1 shown.

[0049] 2. Physiological and biochemical identification

[0050] Refer to "Bergey's Bacterial Identification Handbook" and "Common Bacterial System Identification Handbook" edited by Dong Xiuzhu and Cai Miaoying to conduct physiological and biochemical measurements on the sc...

Embodiment 3

[0062] The cultivation of embodiment 3 bacillus amyloliquefaciens F028

[0063] (1) Selection of medium

[0064] On the basis of 0.8% glucose, 1.5% corn flour, 1.5% soybean flour and 1000mL distilled water in the basic medium, the carbon source, nitrogen source and inorganic salt were optimized and screened by single factor test.

[0065] As carbon source, sucrose, maltose, lactose, soluble starch, mannitol, glycerin, citric acid and dextrin were used instead of glucose. The screening results showed that the strain cultured with mannitol as the carbon source had the highest biomass and the bacterial cell content was the highest. 1.1×10 9 CFU mL -1 , the spore content was 8.9×10 8 CFU mL -1 . Therefore, mannitol was finally determined to be the most suitable carbon source for F028 growth.

[0066] Nitrogen source, peptone, tryptone, urea, ammonium sulfate, ammonium nitrate, potassium nitrate, ammonium chloride and beef extract were added to the basal medium respectively. ...

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Abstract

The invention provides Bacillus amyloliquefaciens F028. The strain is preserved in the China General Microbiological Culture Collection Center on April 20, 2020, and has a preservation registration number of CGMCC No.19590. A biological control preparation prepared from the strain has a broad-spectrum antibacterial effect on bacterial diseases caused by Xanthomonas campestris, Erwiniacarotovora and Ralstonia solanacearum, and fungal diseases caused by Botrytis cinerea, Rhizoctonia solani, AlternariaNees, Fusarium, Phytophthora spp, Glomerellacingulata(Stonem.)Spauld et Schrenk, Colletotrichumcapsici and Beauveria bassiana. According to the invention, a new resource is provided for replacing a chemically synthesized bactericide with microorganisms and preventing and controlling plant diseases by using a biological prevention and control method, and the Bacillus amyloliquefaciens can be developed and utilized as a biopesticide; and moreover, the Bacillus amyloliquefaciens provided by the invention has a promotion effect on the plant growth of tomatoes and tobaccos, and has a wide application prospect.

Description

technical field [0001] The invention belongs to the field of microorganisms, and in particular relates to a strain of bacillus amyloliquefaciens, bacterial agents, preparation methods and applications thereof. Background technique [0002] Walnut bacterial black spot caused by Xanthomonas campestris is one of the main aboveground diseases in walnut production areas in the world. The disease leads to fruit drop, and the fruit drop rate can be as high as 60%, causing serious economic losses. . At present, no walnut variety (clonal line) is completely immune to black spot disease; ginger blast caused by Ralstonia solanacearum mainly damages roots and ginger, and the infected ginger is water-soaked at the beginning , yellowish-brown, gradually softening and rotting inside, the epidemic period of ginger blast is long and the damage is serious. Bacterial soft rot of watermelon caused by Erwinia carotovora is a common disease of melon vegetables and fruits, causing soft rot of th...

Claims

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Application Information

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IPC IPC(8): C12N1/20A01N63/22A01P1/00A01P3/00A01P21/00C12R1/07
CPCC12N1/20A01N63/22
Inventor 李雅华刘新赵方贵赵林刘广超叶青
Owner QINGDAO AGRI UNIV
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