A bacterial strain for degrading neonicotinoid pesticides in soil and its application
A neonicotinoid and soil technology, which is applied in the restoration of contaminated soil, fungi, and microbial-based methods, can solve problems such as acetamiprid residues, and achieve efficient degradation and high repair capabilities.
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Embodiment 1
[0036] Isolation and screening of acetamiprid-degrading strain Fusarium.sp CS-3:
[0037] Take 5 g of acetamiprid-contaminated soil samples and place them in 100 mL of enrichment medium containing 30 mg / L acetamiprid, -1Culture 7d. Use high-performance liquid chromatography (HPLC) to measure the degradation situation of enrichment solution to acetamiprid, after confirming that acetamiprid is degraded, insert in the enrichment medium containing 40mg / L acetamiprid with 5% inoculum size, Continue to enrich and measure the degradation situation, follow this method until the concentration of acetamiprid increases to 100mg / L, and pass passage 3 times.
[0038] The acetamiprid enrichment solution was serially diluted and spread on an inorganic salt plate with 200 mg / L acetamiprid as the sole carbon source, and cultured in a 30°C incubator for 5 days. Single colonies with different colonies grown on the plate were streaked on the LB plate for purification, and inoculated in the liqu...
Embodiment 2
[0041] Identification and growth characteristics of acetamiprid-degrading strain Fusarium.sp CS-3:
[0042] Fusarium.sp CS-3 grows slowly on LB plates, but grows faster on YPD plates. At 30°C, 72 hours can form large and fluffy colonies, dull, dry, rough, opaque, and the vegetative hyphae are tight Based on its 18S rDNA sequence (as shown in SEQ ID No.: 1) and physiological and biochemical characteristics, the strain CS-3 was identified as the Fusarium genus.
[0043] The optimal growth temperature of strain CS-3 is 30°C, and it can grow well at 25°C, but its growth is significantly inhibited at a higher temperature (40°C) ( Figure 2a ). Strain CS-3 grew well at pH 7.0, which was the optimum growth pH; when pH was 4, its growth was significantly inhibited ( Figure 2b ).
Embodiment 3
[0045] Degradation characteristics of acetamiprid-degrading bacteria Fusarium.sp CS-3:
[0046] Pick a single colony of strain CS-3 from the YPD plate, inoculate them in 5mL YPD liquid medium at 30°C, shaker 180r min -1 Cultivate for 48h. Then the culture solution of the bacterial strain was transferred to 100 mL of fresh liquid YPD medium, and the culture was continued for 20 h. 8000r·min -1 Centrifuge for 10 min, collect the bacteria, wash twice with sterilized inorganic salt medium, that is, the seed liquid. In the inorganic salt medium with a final concentration of acetamiprid of 100mg / L, inoculate the seed solution of strain CS-3 according to the inoculation amount of 5% v / v, and incubate at 30°C and 180r min -1 Shake the culture, take samples every 12 hours, and detect the concentration of acetamiprid by HPLC.
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