Nitrogen fixing micrococcus and application thereof
A micrococcus, nitrogenase technology, applied in bacteria, microorganisms, microorganism-based methods, etc., can solve problems such as harm, unbearable for farmers, consumption of large energy, etc., and achieve the effect of broad application prospects.
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Embodiment 1
[0040] Example 1. Isolation and identification of Micrococcus sp. GD192CGMCC No.7778
[0041] 1. Enrichment and isolation of crop rhizosphere nitrogen-fixing bacteria GD192
[0042]Take 10g soil sample (collected from Beijing, China) and shake it in 90ml sterile water for 20 minutes to make a turbid solution, absorb 5ml and put it into 30ml nitrogen-fixing bacteria enrichment culture medium ACCC55, 100rpm, 28℃ for shaking culture, After 72 hours, fresh culture medium was changed to continue cultivation. The nitrogen-fixing spores were isolated after repeating the culture for 3 times. Draw 1ml of the above-mentioned nitrogen-fixing bacteria enrichment culture and put it into 9ml sterile water to make 10 -1 Dilution, continue to dilute to make 10 -2 、10 -3 、10 -4 、10 -5 Bacterial suspensions of dilutions were heated in boiling water at 100°C for 10 minutes, and after cooling, 0.1ml of each dilution was spread on a nitrogen-free medium plate and cultured statically at 29°C....
Embodiment 2
[0072] Example 2, Determination of Nitrogenase Activity of Micrococcus sp. GD192CGMCC No.7778
[0073] Micrococcus (Micrococcus sp.) GD192CGMCC No.7778 obtained in Example 1 is carried out nitrogenase activity assay, and specific method is as follows: in 15 * 150mm screw-top glass tubes, add 5mL improved nitrogen fixation medium to make inclined-plane, inoculate Micrococcus sp. GD192CGMCC No.7778 was cultured at 28°C. Azotobacter chroococcum (Azotobacter chroococcum) ACCC11103, which is commonly used in the production of microbial fertilizers, was used as the positive control, and the blank slant without inoculation was used as the negative control. Three replicates were set for each treatment. After culturing for 72 hours, replace the rubber stopper, inject acetylene gas to make the final concentration 10%, seal it with medical adhesive tape, continue culturing for 72 hours, take 100 μL of reaction gas, measure the amount of ethylene produced by gas chromatography, and calcul...
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