Tebuconazole pesticide-degrading bacteria and soil bioremediation bacteria agent based on the bacteria and its application
A technology of soil biology and restoration of bacterial agents, applied in the biological field, can solve environmental pollution and other problems, achieve the effects of easy use, low production cost, and elimination of pesticide residues
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Embodiment 1
[0026] Example 1: Enrichment, isolation, purification and preparation of bacterial suspension of tebuconazole-degrading bacteria
[0027] Take 5g of soil sample (tebuconazole has been used for many years) and add it to a conical flask filled with 100mL of inorganic salt medium, and take 1mL from the methanol solution of tebuconazole with a concentration of 10000mg / L and directly add it to the conical flask as a microorganism The only carbon source for growth (100mg / L), cultured on a shaker for 7d (30°C, 150r / min) and then transferred to an inorganic salt medium containing tebuconazole 200mg / L, cultured for 7d, and then transferred in turn To culture in the inorganic salt medium containing 300mg / L, 500mg / L, 800mg / L and 900mg / L of tebuconazole, after culturing for 7 days at each drug concentration, then in the plate medium containing 400mg / L of tebuconazole Insert 0.1mL of the above-mentioned culture medium on top, culture in a 30°C incubator for 2 days after coating, pick out a...
Embodiment 2
[0030] Embodiment 2: degradation condition
[0031]1. The effect of pH on the degradation of tebuconazole by strain H2
[0032] It is 4% inoculum amount by volume percentage, the H2 bacterium suspension prepared by embodiment 1 is inserted in the inorganic salt medium that tebuconazole concentration is 200mg / L, regulates the pH of culture medium with hydrochloric acid and sodium hydroxide solution respectively The values were 3, 5, 7, 9, and 11, cultured on a shaker at 30°C and 150 rpm, and the content of tebuconazole was measured at 1, 2, 4, 6, and 8 days, respectively.
[0033] Described inorganic salt medium formula is: K 2 HPO 4 1.5g, KH 2 PO 4 1g,NH 4 NO 3 1g, MgSO 4 ·7H 2 O0.3g, CaCl 2 2H 2 O 25mg, Fe 2 (SO4) 3 8mg, pH natural, 1000mL distilled water.
[0034] The experimental results of pH on the degradation of tebuconazole by strains showed that when the pH was 7.0, the degradation rate reached 91.25% at 6 days, and the degradation efficiency of tebu...
Embodiment 3
[0050] Embodiment 3: the preparation of soil bioremediation bacterial agent
[0051] In the slant test tube strain of tebuconazole-degrading bacteria H2, use an inoculation loop to inoculate a little bacteria on the plate medium, culture at 30°C for 48 hours, then use an inoculation loop to inoculate a little bacteria in LB medium, 30°C Shake culture to the logarithmic phase, the formula of the plate medium is: NaCl 10g, peptone 10g, beef extract 5g, agar 15g, add 1000mL distilled water; the LB medium is: NaCl 10g, yeast extract Powder 5g, tryptone 10g, add 1000mL of distilled water; put 10mL of the cultured strain into the seed bottle, shake culture to the logarithmic phase; 1000mL seed bottle medium is: sucrose 2g, K 2 HPO 4 1.5g, KH 2 PO 4 1g, MgSO 4 ·7H 2 O 0.3g, NH 4 NO 3 1g, CaCl 2 2H 2 O 25mg, Fe 2 (SO4) 3 8mg, pH 7.0, 1000mL distilled water.
[0052] Add 15mL of the cultured strains above to 1000g of fermentation medium consisting of wheat bran, polyvin...
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