A kind of decolorizing bacillus and its application in antagonizing fruit and vegetable fungal diseases
A bacillus and decolorization technology, applied in the field of microorganisms, can solve the problems of crop quality decline, food safety and environmental pollution such as fruits and vegetables
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Embodiment 1
[0026] Embodiment one: separation and purification method
[0027] The separation and purification method of decolorizing bacillus of the present invention, the steps are as follows:
[0028] S1: Weigh 10g of soil samples from the Mangrove National Wetland Park in Hailing Island, Guangdong Province; then, add 90ml of sterile water to the soil samples in the ultra-clean workbench, place on a shaker and vibrate for 60min , so that the soil sample is evenly dispersed in the diluent to become a soil suspension; after the soil is dispersed, draw 100ul of the soil suspension into 900ul of sterile water to obtain a 10-fold dilution, and then dilute 10 times in turn to obtain 10 2 Double dilution, 10 3 Double dilution, 10 4 Double dilution, 10 5 double dilution and 10 6 Double dilution, the whole process is carried out in the ultra-clean workbench.
[0029] S2: Take 100ul of 10-fold dilution, 10 2 Double dilution, 10 3 Double dilution, 10 4 Double dilution, 10 5 double diluti...
Embodiment 2
[0034] Embodiment two: cultivation method
[0035] S1 inoculation: the seed solution obtained in Example 1 was mixed with 2×10 6 Individual / ml inoculum concentration, 2% bacterium liquid and culture medium volume ratio inoculum quantity are inoculated in the 1000ml Erlenmeyer flask that contains 300ml depigmentation Bacillus culture medium;
[0036] S2 culture: culture the culture medium inoculated with the Bacillus depigmentosa at 28° C. and 180 rpm on a constant-temperature shaker for 48 hours to prepare the Bacillus decolorizai bacteria liquid described in the present invention.
[0037]Since the decolorizing bacillus described in the present invention can compete with pathogenic bacteria for nutrients on the one hand, and compete with pathogenic bacteria for binding sites on the plant surface during its own growth process, thereby reducing the pathogenicity of pathogenic bacteria; on the other hand, the present invention The decolorizing bacillus can secrete antibacterial...
Embodiment 3
[0039] Example 3: PCR amplification of 16S rRNA sequence and sequencing
[0040] S1: Extraction of Genomic DNA
[0041] Genomic DNA was extracted using the Omega Bacterial DNA Kit (D3350-01) kit. First, take 2ml of the seed solution described in Example 1 in a sterile 2ml centrifuge tube, centrifuge at 12000rpm for 2min, discard the supernatant and keep the precipitate; then, add 100ul 1×TE Buffer to the precipitate, vortex to mix, add Mix 10ul lysozyme in water bath at 37°C for 10min; add 100ul BTL Buffer and 20ul proteinase K, mix well, incubate at 55°C for 1 hour, shake and mix three times in the middle; add 5ul RNaseA enzyme, mix well, let stand at room temperature for 5min, then centrifuge at 10000rpm for 2min , transfer 200ul of the supernatant to a new sterile 1.5ml centrifuge tube; add 200ul of BTL Buffer, mix well, and place in a warm bath at 65°C for 10min; add 200ul of absolute ethanol, vortex and mix, and transfer all the samples to In the adsorption column, cent...
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