Bacillus velezensis and application thereof in coconut disease control
A technology of Bacillus Velez and Bacillus, applied in the direction of application, bacteria, fungicides, etc., to achieve the effect of strong selectivity and less resistance to drugs
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Embodiment 1
[0026] Example 1 Isolation and screening of strain X1
[0027] (1) Isolation and purification of strains: obtained by dilution coating method. Select 1 g of healthy coconut garden rhizosphere soil, pass through a 60-mesh sieve, and sieve the soil sample according to 10, 10 2 , 10 3 , 10 4 0.1 mL of each gradient soil dilution solution was applied to the LB medium plate with a pipette, and incubated at 30 °C for 4 d. Single colonies were picked according to the different shapes and colors of colonies growing on the plate, and after repeated streaking and purification, they were transferred to the slant of LB medium and stored at 4°C.
[0028] (2) Preparation of the fermentation broth of the strain to be tested: Inoculate the pure culture of the strain to be tested in a 250 mL conical flask containing 100 mL of LB broth, and culture with constant temperature shaking at 160 r / min and 28°C for 1-2 d.
[0029] (3) Screening of antagonistic strains: using the confrontation cultu...
Embodiment 2
[0033] Example 2 Identification of strain X1
[0034] (1) Observation of the morphology and culture characteristics of the strain
[0035] After inoculating strain X1 on LB solid medium for 2 days at 28°C, the colony is nearly circular, with a smooth and moist surface, irregular edges, milky white or pale yellow (see figure 2 ), opaque, no pigment; under the microscope, the cells are rod-shaped, have spores, and are Gram-positive.
[0036] (2) Physiological and biochemical properties of X1
[0037] The physiological and biochemical identification of bacterial strain X1 mainly refers to the "Common Bacterial System Identification Manual" and "Berger Bacterial Identification Manual", and the identification results are shown in Table 2. The physiological and biochemical characteristics of the X1 strain are in line with the characteristics of Bacillus, which is similar to that of Velez. The type strain of Bacillus velezensis is consistent.
[0038] Table 2 Physiological and bioc...
Embodiment 3
[0044] Example 3 Strain X1 fermentation process
[0045] (1) Strain activation. Picked strains were streaked and inoculated onto LB medium plates, cultured in a constant temperature incubator at 30°C for 24 hours, and when a large number of bacterial cells grew on the plates, they were used to transfer liquid seeds.
[0046] (2) Liquid seed preparation. The activated strains were picked and inoculated into the LB medium, shaken at 30° C., 220 r / min for 28 hours, and a large number of bacterial cells were grown to obtain liquid seeds.
[0047] (3) Liquid fermentation. The liquid seeds prepared in step (2) were inserted into the fermentation medium according to the volume ratio of 5% inoculum, the rotation speed was 180-220r / min, the dissolved oxygen was controlled at 30%, the pH was 7.0-7.5, the temperature was 28°C, and the aeration was cultivated for 72h.
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