Application of Mucor circinelloides in degradation of lambda-cyhalothrin
A technology of lambda-cyhalothrin and Mucor crimperides, applied in the fields of application, restoration of polluted soil, organic fertilizer, etc., to achieve the effect of wide application value, simple cultivation conditions and fast reproduction
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Embodiment 1
[0026] Embodiment one: the separation of Mucor circiniferus
[0027] 1. Separation medium:
[0028] Basal medium (per liter): (NH 4 ) 2 SO 4 2g, MgSO 4 ·7H 2 O 0.2g, NaH 2 PO 4 ·H 2 O 0.5g, CaCl 2 2H 2 O 0.1g, K 2 HPO 40.5g, agar 15g, PH7.0
[0029] Separation medium: Sterilize the basic medium at 121°C for 20 minutes, then cool the medium to about 50°C, add lambda-cyhalothrin mother solution under sterile conditions to a final concentration of 500 mg / L, and add ampicillin to The final concentration is 100mg / L, which is the fungal isolation medium.
[0030] 2. Separation method: Take 5g of cotton straw crushed to about 1cm, add 50mL of sterile water, place it on a shaker, shake it at 25°C and 100rpm for 30min, and filter it with a layer of sterile gauze under sterile conditions Mix the solution, discard the filter residue, centrifuge the filtrate at room temperature, 4000rmp for 1min, discard the supernatant, suspend the precipitate with 5mL sterile water, conti...
Embodiment 2
[0031] Embodiment two: the isolation and identification of Mucor circiniferus:
[0032] 1. Sequence determination and analysis of Mucor circinelloides CSGW1 16S rDNA:
[0033] (1) Extraction of PCR template DNA
[0034] Mucor circinelloides CSGW1 was inoculated in PDA liquid medium, cultured at 30°C for 72 hours to obtain bacterial cells, and genomic DNA was extracted using a new plant genomic DNA rapid extraction kit.
[0035] (2) PCR amplification
[0036] Primers:
[0037] ITS1: 5'-TCCGTAGGTGAACCTGCGG-3';
[0038] ITS4: 5'-TCCTCCGCTTATTGATATGC-3'.
[0039] reaction system:
[0040]
[0041]
[0042] The PCR amplification conditions were: pre-denaturation at 94°C for 5min; denaturation at 94°C for 30s, annealing at 54°C for 30s, extension at 72°C for 50s, 35 cycles; extension at 72°C for 10min.
[0043] (3) Sequence determination
[0044] The PCR amplification product is sequenced after electrophoresis detection and purification, and its sequence is shown in SEQ...
Embodiment 3
[0045] Example 3: Degradation of lambda-cyhalothrin by Mucor circinelloides
[0046] 1. Preparation of seed solution
[0047] (1) Strain cultivation: the isolated strain Mucor circinelloides CSGW1 was inoculated in liquid PDA medium, and cultured in a shaker at 30°C with a rotation speed of 120rpm for 3-5 days to the stationary phase. Contains about 15g of dry bacteria per liter;
[0048] (2) Preparation of liquid seeds: take the cultured bacterial solution in step (1), centrifuge at room temperature, discard the supernatant, settle, suspend fully with sterile water, and centrifuge again. After repeating this step several times, use sterile Bacterial water suspends the precipitate to prepare liquid seeds;
[0049] 2. Degradation of lambda-cyhalothrin: select a liquid separation medium, insert the liquid seeds prepared in step (2) under sterile conditions, culture at 30° C. and a rotation speed of 120 rpm for 7 days, and repeat this degradation step.
[0050] The invention p...
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