Preparation method and application of Burkholderia and its metabolites
A technology of Burkholderia and Holderia, which is applied in the field of preparation of Burkholderia and its metabolites, can solve economic losses, chemical fungicides pollute the environment, affect the quality of agricultural products and Quantity and other issues, to achieve the effect of broad antibacterial spectrum
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Embodiment 1
[0066] (1) Separation and purification of strains
[0067] Put the root soil containing the root system into a 45ml concentration of 0.25 % sterile Ringer washing bottle (0.25 % of the concentration of NACL in Ringer washing solution at 7.5g / L, KCL is 0.35g / L, CACL 2 The concentration is 0.21g / L, the same below), and shakes 30 minutes under the conditions of 30 ° C and 180R / min; after static, the upper clearing is diluted with 0.25 % Ringer washing solution to 10 to 10 to 10 -3 10 -4 10 -5 Take 200 μL coating tablets and repeat each gradient; after the colonies occur, use the vaccination needle to pick the smooth colonies of the edge to separate the plate scratch to obtain a pure culture -strain Z1.
[0068] (2) The entire genome sequence and sequence analysis of the strain
[0069] The single bacteria of the strain Z1 were inoculated in the nutritional broth yeast cream medium (NBY medium). After training 2D, collect the bacteria, washed the sterile distilled water once, and sus...
Embodiment 2
[0079] The formula of the liquid fermentation culture group is: 10g / L, beef cream 3g / L, yeast cream 5g / L, sodium chloride 5g / L, pH = 7.5;
[0080] (1) After activating Berke Holder Ceramus, inoculate at a volume of 500ml of liquid fermentation medium at a volume of 5 volumes, and after 72H at the condition of 30 ° C and a speed of 190r / min, add 10ml to add 10ml Large pore resin XAD-16, continue to cultivate 24h to get fermentation liquid;
[0081] (2) Collect the fermented liquid from ingot and large-hole resin XAD-16, abandon the clearing, and get the solid mixture;
[0082] (3) After mixing the solid mixture with a volume of methanol of the equal volume 30min, separate the extraction solution, the extract solution is concentrated through the rotation and evaporation instrument, and then dissolved in 1ml methanol, and then filtered after the speed of 9000r / min 10min. The metabolites of the strain Z1.
Embodiment 3
[0084] The formula of the liquid fermentation culture group is: protein 胨 8g / L, beef cream 2g / L, yeast paste 3g / L, sodium chloride 3g / L, pH = 7;
[0085] (1) After activating Berke Holder Ceramus, inoculate at a volume of 3 % to 500ml of liquid fermentation medium, and after 84H at the condition that the temperature is 25 ° C and the speed of 180r / min, add 5ml to add 5ml Large hole resin ADS-17, continue to cultivate 30h to get fermentation liquid;
[0086] (2) Collect the fermented liquid aggressive and large-hole resin ADS-17, abandon the clearing, and get the solid mixture;
[0087] (3) After mixed with a volume ratio of 1: 2 for 30 minutes, the solid mixture and methanol are separated by the volume ratio, and the extract solution is separated. Filter, get the metabolites of the strain Z1.
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Abstract
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