Nitrate reducing bacterium and application thereof
A technology of nitrate and microbial strains, applied to nitrate-reducing bacteria and its application fields, can solve problems such as secondary pollution, and achieve the effects of reducing crop nitrogen content, soil nitrate nitrogen content, and soil nitrogen loss
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Embodiment 1
[0010] Example 1: Cultivation, isolation, screening and identification of Bacillus Nitratireductor lacus GSS14
[0011] 1.1 Culture and isolation of strains
[0012] The bacillus N.lacus GSS14 of the present invention is obtained by separating and purifying the bottom mud of Yuncheng Salt Lake in Shanxi.
[0013] Concrete separation and purification method comprises the following steps:
[0014] (1) Take 5.0g of Shanxi Yuncheng Salt Lake bottom mud and inoculate it into the enrichment and separation medium with a pH value of 7.2, culture at 30°C with shaking at 200rpm / min; after culturing for 3 days, transfer the culture medium to Transfer to another fresh enrichment and separation medium, and culture with shaking at 30°C and 200rpm / min.
[0015] (2) After four generations of enrichment culture, the culture solution of the fourth generation was diluted to 10 by the dilution plate method. -5 -10 -6 , spread 0.1 mL of the diluted culture solution evenly on the agar solid med...
Embodiment 2
[0031] Embodiment 2: Nitrate reducing effect of nitrate reducing bacteria N.lacus GSS14
[0032] (1) Pick a ring of bacterial lawn from the inclined surface of N.lacus GSS14 and put it into an Erlenmeyer flask equipped with LB liquid medium, and cultivate it on a shaking table at 37°C and 200rpm / min for 36h, and the fermented liquid is the seed liquid.
[0033] Glucose, sodium acetate, sodium lactate and other carbon sources were added to the basic culture of Giltay, and the C / N ratio in the culture medium was adjusted to 3:1. Divide the above culture solution into 500mL Erlenmeyer flasks, 100mL per bottle, inoculate 1mL of N.lacus GSS14 inoculum solution, culture in sequence batch at 37°C, 200rpm / min shaker, detect nitrate nitrogen and ammonia in the culture solution nitrogen content. Ammonia production rate = (NH 4 -N content / NO 3 -N initial concentration) / NO 3 -N initial concentration ×100.
[0034] The formula of the 2216E liquid medium is: yeast powder 1.0g, peptone 5...
Embodiment 3
[0037] Embodiment 3: the preparation of microbial bacterial agent
[0038] (1) Shake flask culture: Pick a ring of bacterial lawn from the N.lacus GSS14 slant and transfer it into a conical flask filled with 100mL 2216E liquid medium, culture it on a shaker at 37°C and 200rpm / min for 24h. The formula of the 2216E liquid medium described is: yeast powder 1.0g, peptone 5.0g, ferric citrate 0.1g, NaCl 19.45g, MgCl 2 5.98g, Na 2 SO 4 3.24g, CaCl 2 1.8g, KCl 0.55g, Na 2 CO 3 0.16g, BrCl 2 0.08g, SeCl 2 0.034g, H 3 BO 3 0.022g, Na 2 SiO 3 0.004g, NaF 0.0024g, NH 4 NO 3 0.0016g, Na 2 HPO 4 0.008g, water 1000mL, pH 7.2.
[0039] (2) Fermentation in seed tank: put the cultured N.lacus GSS14 fermentation broth into a 10L fermenter, and ferment at 37°C and 200 rpm / min for 36 hours; the formula of the fermentation medium used is the above-mentioned liquid medium.
[0040] (3) Fermentation in a fermenter: put the cultured N.lacus GSS14 fermentation broth into a fer...
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