Enterobacter cloacae and application thereof in degradation of thiamethoxam
A technology of Enterobacter cloacae and thiamethoxam, applied in the field of microbiology and food safety, can solve the problems of plant adaptation or different sensitivity, and achieve the effect of reducing absorption and transfer
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Embodiment 1
[0026] Example 1 Isolation, identification and degradation function verification of Enterobacter cloacae TMX-6
[0027] 1. Strains Enterobacter cloacae sp.TMX-6 isolation and identification
[0028] The applicant collected the wild weed Radix Ophiopogon japonicus from the ditch (E120°11, N30°18′) of the former pesticide factory site of Zhejiang Academy of Agricultural Sciences in July 2016, then sterilized the surface of the Ophiopogon japonicus plants, and ground them into juice. A plant endophyte strain with thiamethoxam degrading properties was obtained by coating on the inorganic salt solid medium containing thiamethoxam as the only carbon and nitrogen source. The applicant named the bacterium as TMX-6, and its electron microscope pictures are as follows: figure 1 As shown, the strain was rod-shaped, without endophytic spores.
[0029] The TMX-6 strain was identified as Enterobacter cloacae ( Enterobacter cloacae ), the applicant deposited it on August 8, 2018 at th...
Embodiment 2
[0035] The preparation of embodiment 2 TMX-6 microbial agent
[0036] In the present embodiment, TMX-6 bacterial agent obtains like this:
[0037] A) Inoculate the strain TMX-6 obtained in Example 1 into LB medium, streak at 30°C, and pick a single colony for two cultures, then pick a single colony and put it in the strain activation medium, 30°C, 150-220 rpm Vibrate on a shaking table for 12-24 hours to obtain activated strains;
[0038] B) Inoculate the activated strain into the fermenter equipped with the seed medium, the inoculum amount is 1% of the volume of the seed medium, at 25-38°C, cultivate in air for 16-24h, and obtain liquid seeds;
[0039] C) Inoculate liquid seeds into a fermenter equipped with seed medium, the inoculum amount is 1% of the volume of the seed medium, and culture at 30-35°C and 200 rpm in the dark until the logarithmic growth phase to obtain a viable cell culture;
[0040] D) Take 50 ml of live bacterial culture and centrifuge at 4 ℃, 5000 rpm f...
Embodiment 3
[0041] Example 3 TMX-6 bacterial agent root irrigation inoculation / foliar spraying to repair thiamethoxam pollution experiment of leafy vegetables
[0042] Sampling the natural soil of farmland, first exposed to the sun and air-dried, and then passed through a 30-mesh sieve to remove oversized stone particles. The soil composition property is pH5.97, organic matter content is 66.8 g / kg dry soil, total nitrogen content is 0.36%, and total carbon content is 49.61%. The total hydrogen content is 5.7174%, the total sulfur content is 0.01%, the oxygen content is 44.37%, the clay content is 2.33%, the silt content is 16.1%, and the sand content is 81.5%.
[0043] 1) Soak all the roots of the small green vegetable Shanghai Qing with three leaves and one heart in the TMX-6 bacterial agent obtained in Example 2 (10 7- 8 cfu / mL) for 24 hours, transplanted to the natural soil of the above-mentioned farmland, and the control group 1 was soaked in physiological saline; after 21 days of cu...
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