Fungi strain for degrading pyrethroid pesticides as well as bacterium agent and applications of fungi strain
A technology of pyrethroids and fungi, which is applied in the field of fungal strains and their agents and applications, can solve the problems of limited number of microbial strains, low degradation efficiency, complex mechanism, etc.
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Embodiment 1
[0038] Embodiment 1: Isolation and purification of Fusarium solani (Fusarium solani) JZB41C002
[0039] Take 3 samples of vegetable field soil samples from Tongzhou District, Beijing, where beta-cypermethrin has been applied, mix them evenly, take 10g each, and add them to 100mL enrichment culture containing beta-cypermethrin at a concentration of 50mg / L under aseptic conditions. In the Erlenmeyer flask of base, cultivate 7d on 120r / min shaker under 30 ℃; Then press 10% inoculum amount ("10% inoculum amount" in this embodiment refers to the volume ratio of inoculum and inoculated medium) Transfer it to the next batch of enrichment medium containing 100 mg / L of beta-cypermethrin, and continue to cultivate for 7 days; transfer to the enrichment medium containing 200 mg / L of beta-cypermethrin according to the inoculum size of 10% in, cultivated for 7d; then transferred to the basic inorganic salt medium containing beta-cypermethrin concentration of 200mg / L by 10% inoculum size, a...
Embodiment 2
[0040] Example 2: ITS sequence identification of Fusarium solani (Fusarium solani) JZB41C002
[0041] The acquisition of PCR templates includes two methods, one is to extract genomic DNA and use it as a PCR template, and the other is to use a special enzyme from Takara Company to directly perform fungal colony PCR. Both methods are described below.
[0042] Universal primers for fungal identification, namely ITS1: 5'-TCC GTA GGT GAA CCT GCG G-3' (SEQ ID No.2), ITS4: 5'-TCC TCC GCT TAT TGA TAT GC-3' (SEQ ID No. 3).
[0043] A) Direct colony PCR:
[0044] a) Template preparation: Inoculate the purified fungus JZB41C002 of the present invention on PDA medium, culture in a constant temperature incubator at 25-28°C for 2-3 days, and pick a very small amount of hyphae as a template for PCR amplification. This embodiment adopts this method to obtain a PCR template.
[0045] b) PCR reaction system: Mighty Amp DNA Polymerase (purchased from Takara Company) 0.5 μL, ITS1 (10 μmol / L) ...
Embodiment 3
[0054] Embodiment 3: Degradation effect determination of bacterial strain JZB41C002 to beta-cypermethrin
[0055] 1) Cultivate the purified bacterial strain JZB41C002 in LB liquid medium to the logarithmic period (the logarithmic period is determined by measuring the OD600 value, and the OD600 value reaches 0.8, which is the logarithmic period), and then with 10% inoculation amount (this implementation "10% inoculum size" in the example refers to the volume ratio of the inoculum and the inoculated medium) it is inserted into the basic inorganic salt culture solution containing beta-cypermethrin concentration of 50mg / L, at 30 ° C, 120r / min Shaking culture was carried out on a constant temperature shaker for 5 days, and the culture solution without bacteria was set as a control, and each sample was treated with 3 replicates. After the cultivation, absorb 2 mL of the supernatant, add 10 mL of acetonitrile, 2 g of NaCl, vortex for 1 min, let stand for 30 min, separate layers, abso...
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