High-efficiency prothioconazole-degrading bacteria w313, bacterial agent and its application
A technology for efficiently degrading bacteria and prothioconazole, applied in the directions of bacteria, hydrolytic enzymes, microorganisms, etc., can solve problems such as prothioconazole residues
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Embodiment 1
[0049] Isolation and screening of bacterial strains: The activated sludge was collected from the wastewater treatment pond of a pesticide factory containing prothioconazole, and the continuous enrichment culture method was used to take 5mL of culture solution to 100mL of inorganic salts with prothioconazole as the only carbon source In liquid medium (K 2 HPO 4 ·3H 2 O 1.00g / L, MgSO 4 ·7H 2 O 2.00g / L, CaSO 4 0.40g / L, NaCl 0.50g / L, FeSO 4 ·7H 2 O0.01g / L, (NH 4 ) 2 SO 4 1.00g / L), cultivated in a constant temperature incubator at 30°C for 48h; inoculated the culture solution in the medium containing 700mg / L prothioconazole inorganic salts on the solid medium, separated and purified, and obtained a strain of prothioconazole Conazole-efficient degrading bacteria, named W313, were preserved on the slant.
Embodiment 2W313
[0050] Identification of embodiment 2W313 bacterial strain
[0051] 1. Morphological identification: The preliminary morphological identification of strains is based on the "Bacteria Identification Manual"; figure 1 As shown, the W313 strain grows slowly on the inorganic salt medium, and the surface of the colony is flat and wet, gray-green; the W313 strain can grow in the medium with prothioconazole as the only carbon source, and its resistance to The higher tolerance and utilization rate indicate that it has higher prothioconazole degradation activity. The results of Gram staining of W313 strain were as follows: figure 2 shown.
[0052] 2. Molecular biological identification of the strain: DNA extraction, PCR amplification and sequence analysis of the amplified product of the W313 strain: the fresh bacteria cultured for 4 days were used as the DNA extraction material, and the genomic DNA of the W313 strain was extracted by urea extraction method, 0.8% agar The purity was...
Embodiment 3
[0057] Tolerance test of W313 strain to prothioconazole on the plate:
[0058] The W313 strain was inoculated on a solid plate with prothioconazole as the only nutrient source, and the concentrations of prothioconazole were 10mg / L, 50mg / L, 100mg / L, and 200mg / L respectively. Conazole concentration is 200mg / L culture medium grows normally, the result is as follows Figure 4 with Figure 5 as shown, Figure 4 The concentration of prothioconazole in the medium is 20ppm, Figure 5 The concentration of prothioconazole in the medium was 200ppm, indicating that the tolerance of the bacteria to prothioconazole was high.
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