Bacillus pumilus and application thereof
A technology of bacillus pumilus and bacterial agent, applied in the field of microorganisms, to achieve the effect of broad antibacterial spectrum, reduced application, and good biocontrol effect
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Embodiment 1
[0037] Embodiment 1: Isolation and screening method of Bacillus pumilus GLB197
[0038] (1) Sample collection:
[0039] Sample collection: Samples were collected from the leaves of the main grape varieties in 13 major grape producing areas across the country, stored in a low-temperature box and brought back to the laboratory. The isolation of endophytic bacteria was completed within 24 hours. The location and time of sample collection are shown in Table 1.
[0040] Table 1 Sample collection location and time
[0041]
[0042]
[0043] (2) Obtaining of Bacillus pumilus GLB197 strain:
[0044] 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 four times with sterile water. Take 100 μl of the fourth rinsing water to coat the TSA plate, set up 3 replicates, and culture in a constant temperature incubator at 28°C for 4 to...
Embodiment 2
[0053] Example 2: Antagonism profile determination of Bacillus pumilus GLB197
[0054] 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 pumilus GLB197 had strong antagonistic activity against grape, apple, wheat and other plant pathogenic bacteria. The antibacterial spectrum of Bacillus pumilus GLB197 strain is shown in Table 2.
[0055] Table 2 Antibacterial spectrum of Bacillus pumilus GLB197
[0056]
[0057]
[0058] Among them, +, ++ and +++ respectively represent the three grades of antibacterial effect in PDA medium, namely 31-50%, 51-70% and ...
Embodiment 3
[0059] Embodiment 3: Laboratory control experiment of Bacillus pumilus GLB197 to grape downy mildew
[0060] 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 pumilus GLB197 was inoculated into LB liquid medium, and was shaken at 180 rpm 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 ...
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