Bacillus subtilis FC12 for degrading fluoroglycofen-ethyl and application thereof
A technology of acifluorfen and Bacillus, which is applied in the biological high field, can solve problems such as residues, achieves low production and use costs, solves the problems of excessive pesticide residues and herbicide phytotoxicity, and improves added value.
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Embodiment 1
[0025] Example 1 Screening and isolation of acifluorfen-degrading Bacillus FC12
[0026] Get 3.0ml of activated sludge obtained from the waste water treatment tank of a certain pesticide factory production workshop that produces acifluorfen, and add it into 100ml inorganic salt liquid culture that the concentration of acifluorfen is 100mg / L, and its formula is: Liter contains 1.50 g K 2 HPO 4 , 0.50 g KH 2 PO 4 , 0.20 g MgSO 4 ﹒ 7H 2 O, 1.00 g NaCl, 1.00 g (NH 4 ) 2 SO 4 , pH 7.0, cultured with shaking at 160r / min, 30°C, transferred to fresh inorganic salt medium every 5 days at 3% inoculum, and transferred 5 times in a row.
[0027] Take 1.0ml of the enriched bacteria solution obtained above, add 9.0ml of sterile water, and make 10 -1 The rich solution, and then draw 1.0ml prepared 10 -1 The enrichment solution was added to 9.0ml sterile water, mixed thoroughly to make 10 -2 The enrichment solution, and so on, carry out gradient dilution on the enrichment solution...
Embodiment 2
[0029] Example 2 Shake Flask Degradation Experiment of Bacillus FC12
[0030] In the inorganic salt medium containing 100 mg / L acifluorfen, the formula is: 1.50 g K per liter 2 HPO 4 , 0.50g KH 2 PO 4 , 0.20g MgSO 4 ﹒ 7H 2 O, 1.00g NaCl, 1.00g (NH 4 ) 2 SO 4 , pH 7.0, inoculate YS-03 according to 3% inoculum size, culture with shaking at 30°C, and take samples for measurement after 48 hours. Compared with the blank, by image 3 It can be seen that the absorption peak of the treated sample decreased significantly, and fluorfen was degraded by more than 90%.
Embodiment 3
[0031] Embodiment 3 Containing the preparation of bacillus FC12 bacterial agent
[0032] The original species of acifluorfen-degrading Bacillus FC12 screened and isolated in Example 1 were activated on a petri dish, and the degradation performance was measured, and then inoculated on the inclined surface of a test tube for later use. The test tube was inoculated in a 1000ml shake flask containing 200ml LB medium. The formula of LB medium was: yeast extract 5.00g / L, peptone 10.00g / L, NaCl 10.00g / L, pH 7.0, constant temperature shaking culture to logarithmic phase , ready to inoculate the seed pot. The seed tank is 500 liters, the feeding volume is 400 liters, the medium formula is: glucose 0.8%, (NH 4 ) 2 SO 4 1%, K 2 HPO 4 0.2%, MgSO 4 0.05%, NaCl 0.01%, CaCO 3 0.3%, yeast extract 0.02%, pH 7.2-7.5. After the feeding is completed, 121°C high-pressure damp heat sterilization, after cooling to 33°C, inoculate the above-mentioned cultivated shake flask strains into a ...
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