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Bacillus subtillis and application thereof

A technology of Bacillus subtilis and bacterial agents, applied in the field of microorganisms, to achieve the effects of broad antibacterial spectrum, reduced application, and good biocontrol effect

Inactive Publication Date: 2016-08-31
CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0007] The present invention aims to solve the problems described above

Method used

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  • Bacillus subtillis and application thereof
  • Bacillus subtillis and application thereof
  • Bacillus subtillis and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] Embodiment 1: Isolation and screening method of Bacillus subtilis GLB191

[0039] (1) Sample collection:

[0040] The samples were collected from the leaves of main grape varieties in 13 major grape producing areas across the country, stored in a low temperature box and brought back to the laboratory, and the isolation of endophytic bacteria was completed within 24 hours. The location and time of sample collection are shown in Table 1.

[0041] Table 1 Sample collection location and time

[0042]

[0043]

[0044] (2) Obtaining of Bacillus subtilis strain GLB191:

[0045] Surface disinfection: take fresh grape leaves, rinse them with sterile water, treat them with 70% ethanol (20s), 2% sodium hypochlorite solution (3min), and 70% ethanol (20s) respectively, then rinse them with sterile water 4 times, Take 100 μl of the fourth rinse water to coat the TSA plate, set up 3 replicates, and culture in a constant temperature incubator at 28°C for 4-5 days. No colony f...

Embodiment 2

[0054] Example 2: Antagonism profile determination of Bacillus subtilis GLB191

[0055] The prepared pathogenic bacteria cakes were inserted into the PDA medium at a distance of 3 cm for confrontation culture, and the bacteria cakes inoculated with each test bacteria were used as a control, and placed in an incubator at 25°C for dark culture, and each treatment was repeated 3 times. After 3 days, the colony diameter was measured, and the inhibition rate of the test strains to the growth of pathogenic bacteria was calculated. The results showed that Bacillus subtilis GLB191 had strong antagonistic activity against various crop pathogenic bacteria such as grape, pear, pepper, potato, strawberry and cotton. The antibacterial spectrum of Bacillus subtilis GLB191 strain is shown in Table 2.

[0056] Table 2 Antibacterial spectrum of Bacillus subtilis GLB191

[0057]

[0058]

[0059] Among them, +, ++ and +++ respectively represent the three grades of antibacterial effect i...

Embodiment 3

[0060] Embodiment 3: Laboratory control experiment of Bacillus subtilis GLB191 to grape downy mildew

[0061] Fresh grape downy mildew diseased leaves were collected from the field, brought back to the laboratory for overnight moisturizing cultivation, and fresh sporangia were washed with a brush to prepare 1 × 10 4 cells / mL suspension for use. Bacillus subtilis GLB191 was inoculated with LB liquid medium, and shaken at 180rpm until more than 95% of the bacteria produced spores. Take fresh grape leaves, beat them into leaf disks with a diameter of 1.5 cm, soak them in a 10-fold dilution of Bacillus suspension for 30 minutes, take them out, and dry them on an ultra-clean workbench. Spread 4 layers of gauze moistened with sterile water in a sterilized empty disk, place the dried leaf disks in the disk, put 30 leaf disks in each disk, seal with plastic wrap to keep moisture, and soak the leaf disks in sterile water The plate was used as a control, and each treatment was set up ...

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Abstract

The invention discloses Bacillus subtillis and application thereof. The Bacillus subtillis GLB191 is preserved in China General Microbiological Culture Collection Center (CGMCC) on Oct 29th, 2014, with an accession number of CGMCC No. 9870. The Bacillus subtillis has a wide antibacterial spectrum. An inoculum produced through fermentation of the Bacillus subtillis in the invention can prevent diseases, protect environment and reduce chemical pesticide residuals, and has wide application prospects in sustainable agricultural development.

Description

technical field [0001] The invention relates to the field of microorganisms, in particular to a Bacillus subtilis and an application thereof. Background technique [0002] Grape is an important fruit tree species in China, and my country's grape planting area and output rank among the top in the world. However, the current situation of planting is not optimistic, especially because there are many types of pests and diseases, serious damage, and the frequency and amount of pesticides used are large. Grape downy mildew is the most serious worldwide disease of table and wine grapes. The pathogen causing grape downy mildew is Plasmoparaviticola (Berk. & Curtis.) Berl et de Toni, which belongs to Flagellate, Peronomyces, and Monopota, and is an obligate parasite. The pathogen mainly infects grape leaves, but can also infect inflorescences and young fruits. [0003] Chemical pesticide control is still the main method to control grape downy mildew at present, and is widely accep...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20A01N63/00A01N63/02A01P3/00C12R1/125
CPCA01N63/00A01N63/10C12N1/205C12R2001/125
Inventor 王琦张珣周莹莹李燕
Owner CHINA AGRI UNIV
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