Biological agents and their application to alleviate cadmium stress in leafy vegetable planting soil
A technology of biological agents and leafy vegetables, applied in the field of biological agents to relieve cadmium stress in leafy vegetable planting soil, to achieve the effect of expanding the range of selenium addition, large application prospects, and strong biological functionality
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Embodiment 1
[0044] A biological agent and application thereof for alleviating cadmium stress in leafy vegetable planting soil, the specific steps are as follows:
[0045] (1) Added 400mmol L after sterilization -1 In the LB culture medium of sodium selenite, inoculate 10% live bacteria body of Bacillus subtilis 168 according to the volume ratio, and then place the culture bottle in a shaking flask incubator at 30°C under the condition of 150 rpm. Incubate with oxygen for 24 hours, then centrifuge at 4000g for 10 minutes, collect the precipitate to obtain a mixture of Bacillus subtilis and its metabolite nano-selenium;
[0046] Wherein, the formula of LB (Luria-Bertani) is: 10g / L tryptone, 5g / L yeast extract and 10g / L sodium chloride, adjust the pH value to 7.0-7.5, and sterilize.
[0047] (2) Wash the resulting precipitate with 0.9% NaCl solution, then add liquid nitrogen and beat it with a pestle to break the precipitate; then ultrasonically treat it at 200W for 5 minutes to make the pa...
Embodiment 2
[0054] A biological agent and application thereof for alleviating cadmium stress in leafy vegetable planting soil, the specific steps are as follows:
[0055] (1) Add 500mmol L after sterilization -1 In the LB culture medium of sodium selenite, inoculate 6% of Bacillus subtilis 168 living thalline according to the volume ratio, then place the culture bottle in a shake flask incubator at 32°C under the condition of 200 rev / min. Incubate with oxygen for 14 hours, then centrifuge at 4500g for 15 minutes, collect the precipitate to obtain a mixture of Bacillus subtilis and its metabolite nano-selenium;
[0056] (2) Wash the resulting precipitate with 0.9% NaCl solution, then add liquid nitrogen and beat it with a pestle to break the precipitate; then ultrasonically treat it at 100W for 10 minutes to make the particle size of the precipitated mixture below 200nm, and use 1.5mol L -1 1. The Tris-HCl buffer solution with pH 8.3 was continuously centrifuged at 10,000 g for 5 minutes ...
Embodiment 3
[0062] The biological preparation prepared by using nano-selenium prepared by Bacillus subtilis 168 biological method according to the present invention is compared with the biological preparation prepared by commercially available nano-selenium in alleviating the effect of cadmium stress in leafy vegetable planting soil
[0063](1) 500 mL fermented nano-selenium nutrient solution (250 mg / L selenium content) was purchased from Dongming Wheat Cooperative, and 133.3 g Bacillus subtilis was added to prepare biological preparations.
[0064] (2) Rapeseed seedlings were raised for two weeks in the early stage, and then transferred to a hydroponic device. Slow the seedlings with 1 / 2 Hoagland nutrient solution for three days, then start to add cadmium-containing solution (50 μmol L -1 ). Afterwards, 8 days after the growth of rape seedlings, the leaves of rapeseed were evenly sprayed for the first time, specifically, 5, 10, 15, and 25 mg / L nano-selenium containing 26.7 mg / L Bacillus...
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