A kind of chloroacetamide herbicide degrading bacterial strain and its production bacterial agent and application
A chloroacetamide and herbicide technology is applied to chloroacetamide herbicide-degrading bacterial strains and their produced inoculants and application fields, and can solve problems such as unfavorable application, long time consumption, low acetochlor degradation rate, and the like, Achieve the effect of protecting physical health, protecting the ecological environment, wide application potential and value
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Embodiment 1
[0030] Embodiment 1, isolation and identification of bacterial strain
[0031] Add 5 g of acetochlor-contaminated soil samples into 100 mL of liquid inorganic salt medium (hereinafter referred to as MM) containing 0.2 mM acetochlor, culture on a shaker at 30°C and 150 rpm for 7 days, and transfer to 5% inoculum size (v / v) In the same fresh medium, the enrichment culture was carried out four times in succession. Dilute and spread the fifth-generation enrichment solution on MM solid medium containing 0.2mM acetochlor, culture at 30°C for 5 days, pick a single colony on the plate, transfer it to a 3mL LB test tube for culture, and collect the bacteria by centrifugation , transferred to 20mL MM medium containing 0.2mM acetochlor, cultured at 30°C for 5 days, freeze-dried the culture medium and redissolved it in 5mL methanol, and detected the degradation of single colony by UV scanning and high performance liquid chromatography (HPLC) Finally, an acetochlor-degrading strain AC-1 w...
Embodiment 2
[0033] Embodiment 2, laboratory degradation experiment
[0034] 2.1 Preparation of seed solution
[0035] Insert the strain AC-1 into 100mL LB medium containing 0.2mM acetochlor, culture on a shaker at 30°C and 150rpm, collect the bacteria by centrifugation at 6000rpm after 24h, wash the bacteria twice with MM, and finally resuspend with 10mL MM , as a seed solution for later use.
[0036]2.2 Degradation of chloroacetamide herbicides by strain AC‐1
[0037] The strain AC‐1 was inoculated into 100mL MM containing 0.2mM alachlor, acetochlor, butachlor and metolachlor at 5% inoculum, cultured on a shaker at 30°C and 150rpm, every 8 Take 5 mL of sample every hour, and take it up to 48 hours. The residues of alachlor, acetochlor, butachlor and metolachlor were detected, the degradation rate was calculated, and the time-degradation curve of strain AC-1 to chloroacetamide herbicides was drawn. Such as figure 2 , the strain AC-1 can completely degrade 0.2mM acetochlor within 48h...
Embodiment 3
[0047] Embodiment 3, soil degradation experiment
[0048] The vegetable garden soil was taken as the test soil sample. Pass the soil sample through a 2mm sieve, take a certain amount of alachlor, acetochlor and butachlor powder and dissolve them in 10mL of methanol, and then soak in diatomaceous earth to completely absorb the pesticide. The soaked diatomaceous earth is placed in a fume hood to dry, and it is mixed into the soil so that the concentration of the pesticide in the soil is about 1mg / kg. 500g of each soil sample was cultured in a constant temperature incubator at 30°C, and the inoculum was inserted into the seed solution at an inoculation amount of 10%, and the soil without inoculation was used as a control, and the water holding capacity of the soil was maintained at 60%. After culturing for 7 days, the residual amount was determined by HPLC. The measurement results are shown in Table 1.
[0049] It can be concluded from Table 1 that after 7 days of cultivation,...
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