Bacillus methylotrophicus 4-L-16 for prevention and control of banana vascular wilt and its application
A technology of banana fusarium wilt and methylotrophic type, which is applied in the application, chemicals for biological control, bacteria and other directions, can solve the problems of narrow spectrum of antagonistic bacteria, insignificant prevention and control effect, and incapability of large-scale colonization, etc. Rapid reproduction, good development and application prospects, and easy colonization and reproduction
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Embodiment 1
[0031] Origin, isolation and identification of strains
[0032] The strain is isolated from rapeseed cake, brown sugar and water fermentation broth. The specific preparation method of the fermentation broth is as follows: dissolve 155g of brown sugar in 1000mL of water, dissolve 95g of rapeseed cake in brown sugar water, and adjust the pH value of brown sugar water to 7.0 , placed in a cool and ventilated place, sealed and stored to obtain an aqueous solution of brown sugar containing cake fertilizer; the aqueous solution of brown sugar containing cake fertilizer was placed in a 5L volumetric flask, and cultured with shaking for 7 days at 180rpm and 30°C, filtered to obtain a reddish-brown solution of Fermented broth.
[0033] 1. Isolation and primary screening of antagonistic bacteria in fermentation broth
[0034] Using the plate dilution coating method, the dilution (10 -1 ~10 -3 ), use PDA medium to isolate fungi; LB medium to isolate bacteria; Gaoshi No. 1 medium to is...
Embodiment 2
[0049] The prevention and control effect of embodiment 2 methylotrophic bacillus 4-L-16 to banana Fusarium wilt
[0050] 2.1 Plate Antagonism Test of Methylotrophic Bacillus 4-L-16 on Banana Fusarium wilt Race 1 FOC1
[0051] Methylotrophic Bacillus 4-L-16 was activated on LB medium, placed in a 37°C incubator for 72 hours, and then a 5mm piece of FOC1 mycelium was placed in the center of the potato dextrose PDA medium plate. Inoculate 5 μL of methylotrophic Bacillus 4-L-16 culture solution at 2.5 cm of FOC1 mycelial block, and culture in a 28°C incubator with a plate not inoculated with methylotrophic Bacillus 4-L-16 as a control After 5-7 days, the width of the measured bacteriostatic zone was 9 mm, and it had obvious inhibitory effect on race 1 of Fusarium wilt bacterium, as shown in Table 3 and figure 1 .
[0052] 2.2 Plate Antagonism Test of Methylotrophic Bacillus 4-L-16 on Banana Fusarium wilt Race 4 FOC4
[0053] Methylotrophic Bacillus 4-L-16 was activated on LB me...
Embodiment 3
[0062] Example 3 Biological preparation containing methylotrophic bacillus 4-L-16
[0063] First, activate the methylotrophic Bacillus 4-L-16 strain prepared in Example 1 on LB medium, place it in a 37°C incubator and cultivate it for 72 hours, then use a pipette to draw the 5~10 μL of the thalline was placed in 100mL of 10% brown sugar aqueous solution by mass percentage, shake cultured, the culture temperature was 30°C, the shaker speed was 180rpm, and the culture time was 3~5 days, and the methyl-containing The seed liquid of the vegetative Bacillus 4-L-16 strain is then added to the mixed solution of rapeseed cake and brown sugar at a ratio of 1:10, and the preparation ratio of the mixed solution is: according to the ratio of rapeseed cake The C / N and nitrogen content of the brown sugar, and the carbon content of the brown sugar, the C / N of the substance in the mixed solution is adjusted to be 15 / 1 by a conventional method, and an appropriate amount of water is added.
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