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Methylotroph bacillus and application thereof

A methylotrophic, bacillus technology, applied in the application, bacteria, fungicides and other directions, can solve the problems of difficult to implement disease resistance breeding, the decline of chemical pesticide control effect, etc., to achieve good development and application prospects, rapid and large-scale colonization, Use a wide range of effects

Active Publication Date: 2017-10-20
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Since Botrytis cinerea is a facultative parasite, there is also a saprophytic stage, but its antigen has not been found so far, making it difficult to implement disease-resistant breeding; agricultural cultivation and control mainly through strengthening water and fertilizer management in the field, adjusting temperature, humidity and Lighting, etc. In this way, complete control facilities for field conditions are required, which have not been realized under the existing conditions of strawberry growers in my country; at present, chemical pesticide control is still the main means of preventing and treating strawberry gray mold in my country, but due to continuous use Chemical agents, especially the continuous use of a single agent, lead to the gradual development of drug resistance of Botrytis cinerea, and the control effect of chemical pesticides is also declining year by year. In addition, pesticide spraying cannot avoid direct spraying on the fruit surface when applying pesticides. Chemicals also caused pesticide contamination of fruit

Method used

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  • Methylotroph bacillus and application thereof
  • Methylotroph bacillus and application thereof
  • Methylotroph bacillus and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] 1 Materials and methods

[0025] 1.1 Medium and reagents

[0026] Beef extract-peptone medium: 0.3g beef extract, 0.5g peptone, 0.3g NaCl, 2g agar, 100mL water, pH7.2-7.4, used for the cultivation of antagonistic bacteria.

[0027] PDA medium: 20g potato, 2g glucose, 2g agar, 100mL water, natural pH. For the cultivation of pathogenic fungi.

[0028] Enrichment medium: 200g potato, 10g glucose, 3g beef extract, 5g peptone, 5g NaCl, KH 2 PO 4 2g, MgSO 4 1g, 20g agar, 1L water, pH 7.2. Used for confrontation culture and determination of bacteriostatic rate.

[0029] LB liquid medium: peptone 10g / L, sodium chloride 5g / L, yeast extract 10g / L, solvent is distilled water, pH value 7.0. For seed liquid preparation.

[0030] 1.2 Breeding of dominant degrading strains

[0031] 1.2.1 Source of strain

[0032] The rhizosphere soil of strawberry plants was collected in Fenghua Strawberry Greenhouse, Ningbo City, Zhejiang Province.

[0033] 1.2.2 Isolation, purification an...

Embodiment 2

[0052] Antagonism and bacteriostatic rate of embodiment 2 bacterial strain LM-N to Botrytis cinerea, Rhizoctonia solani, wheat scab

[0053] determination

[0054] Strawberry Botrytis cinerea, wheat head blight and Rhizoctonia solani described in this embodiment were provided by the Plant Pathology Laboratory of Zhejiang University College of Agriculture and Biotechnology (conventional public knowledge).

[0055] For testing pathogenic fungi (Botrytis cinerea, Rhizoctonia solani, Fusarium head blight, commercially available, common pathogenic bacteria preserved in various plant pathology laboratories) in Activate on a PDA plate at 28°C for 48 hours, use a puncher (diameter 6mm) to prepare a bacterial cake at a symmetrical position on the edge of the colony, inoculate the mycelium face down on the center of the enrichment medium plate, and cultivate in the dark at 28°C until the diameter of the bacterial cake reaches 2 cm, sterilized single-layer filter paper with the same por...

Embodiment 3

[0059] Example 3 Verification of the field effect of methylotrophic Bacillus CGMCC No.13541 fermentation broth on strawberry gray mold

[0060] 3.1 Drugs to be tested

[0061] Methylotrophic Bacillus CGMCC No.13541 fermentation broth; 50% dihydantoin wettable powder (commercially available).

[0062] Described fermented liquid is prepared as follows:

[0063] (1) Inoculate methylotrophic Bacillus LM-N into beef extract peptone medium, cultivate at 30°C for 20-24h, and obtain slant bacteria; the beef extract peptone medium consists of: beef extract 3g / L, Peptone 5g / L, sodium chloride 3g / L, agar 20g / L, solvent is distilled water, pH value 7.0;

[0064] (2) Pick slant bacteria and inoculate them into LB liquid medium, cultivate them at 30°C for 24 hours to obtain seed liquid; the LB liquid medium consists of: peptone 10g / L, sodium chloride 5g / L, yeast extract 10g / L L, the solvent is distilled water, the pH value is 7.0;

[0065] (3) Fermentation culture: inoculate the seed li...

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Abstract

The invention discloses a methylotroph bacillus and application thereof to preparation of a plant pathogenic bacterium sterilization agent. The methylotroph bacillus CGMCC No.13541 has the antagonism effect on various phytopathogens such as botrytis cinereal, fusarium graminearum and rhizoctonia solani, and belongs to an environment-friendly antagonistic strain with the obvious plant disease prevention and control effect; the development and application prospects are good. Culture liquid obtained through culture by a beef extract liquid culture medium can be used for preventing and controlling plant diseases. The bacteriostasis rate on the botrytis cinereal is as high as 84 percent; the value is about 8 percent higher than that of ordinary biocontrol bacteria; the foundation is laid for the novel disease prevention microbe resource development and utilization. The resistance capability is high; the use range is wide; the growth speed is high; fast and great-amount multiplication can be realized; the popularization and application of a biological prevention and control preparation in the field are convenient.

Description

(1) Technical field [0001] The invention relates to a methylotrophic bacillus (Bacillus methylotrophicus) LM-N isolated from the soil of a strawberry greenhouse and an application thereof in preventing and treating gray mold of strawberries. (2) Background technology [0002] Botrytis spp. caused by Botrytis spp. is an important class of plant diseases, and it is more common on fruits and vegetables such as strawberries, grapes, tomatoes, lettuces, and cucumbers. The onset of different stages may cause damping-off of plants at the seedling stage or rot of flowers and fruits. It has developed into one of the important diseases that affect the production or storage of vegetables, fruits and flowers in protected areas. Botrytis cinerea can reduce the output of vegetables in protected areas by more than 20%, and serious areas with improper control of disease sources or cultivation measures may even face extinction. In my country, strawberry has been listed as one of the ten pil...

Claims

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Application Information

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IPC IPC(8): C12N1/20A01N63/02A01P3/00C12R1/07
CPCA01N63/10C12N1/205C12R2001/07
Inventor 谢晓梅廖敏吕婷方至萍
Owner ZHEJIANG UNIV
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