Acinetobacter with authigenic nitrogen fixation capacity and application thereof
A bacillus and nitrogen fixation technology, applied in the direction of microorganism-based methods, bacteria, microorganisms, etc., can solve the problems of inability to meet the requirements of polluted soil remediation, resistance to heavy metal growth and nitrogen fixation, and slow growth.
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Embodiment 1
[0061] According to the screening method of the nitrogen-fixing bacterial strain provided by the present invention, a nitrogen-fixing bacterial strain with high nitrogenase activity capable of tolerance to heavy metal cadmium is selected, and the preparation steps are as follows:
[0062] 1) Use the autogenous nitrogen-fixing bacterium Acinetobacter YSGD07 screened from the plant rhizosphere in the laboratory as the starting strain;
[0063] 2) Mutation breeding
[0064] (1) Prepare the single cell suspension of the starting strain YSGD07
[0065] The starting strain YSGD07 was inoculated in liquid medium A, cultured at 28°C, 150rpm for 16hrs, centrifuged, washed with sterile saline, placed in a conical flask with glass beads, and oscillated to disperse into a single-cell bacterial suspension liquid;
[0066] The composition of the medium A is: yeast powder 10g, NaCl 5g, peptone 10g, FeSO 4 ·7H 2 O 0.005g, Na 2 MoO 4 2H 2 O 0.0025g, distilled water 1000ml, pH 7.0~7.2.
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Embodiment 2
[0078] According to the method for preparing nitrogen-fixing bacterial agent provided by the present invention, the nitrogen-fixing bacterial agent with high activity of nitrogenase that can tolerate heavy metal cadmium is prepared, and the steps of preparation are as follows:
[0079] The preparation of bacterial agent comprises the following steps:
[0080] 1. Strain fermentation
[0081] ①First-level Erlenmeyer flask liquid culture
[0082] The frozen Ymu7-3 was quickly thawed at 37°C, and transferred into the Erlenmeyer flask containing liquid medium A according to the inoculum amount of 0.5%, and cultured at 28°C with shaking at 150rpm for 18hrs;
[0083] The composition of the liquid medium A is: the composition of the medium A is: yeast powder 10g, NaCl 5g, peptone 10g, FeSO 4 ·7H 2 O 0.005g, Na 2 MoO 4 2H 2 O 0.0025g, distilled water 1000ml, pH 7.0~7.2
[0084] ②Secondary fermentation tank fermentation
[0085] 1) Put the primary culture medium of Ymu7-3 into ...
Embodiment 3
[0090] Use embodiment 2 bacterial agents to repair cadmium-contaminated soil in strengthened Indian mustard, and its specific steps are:
[0091] The potting soil was collected from the farmland soil in Chang’an County, Shaanxi Province. The total Cd content of the tested soil was 0.25 mg / kg, and 4 soils were added with Cd 2+ levels, respectively 0, 20, 40, 80mg / kg, the corresponding amount of CdCl 2 Dubbed into a solution, so that the test soil and heavy metals are repeatedly mixed evenly, and left to balance for 30 days. In the experiment, a blank control group, a urea-added group and a bacterial agent-added group were respectively set up. After emergence, keep 3 plants in each pot. Dilute the liquid bacterial agent prepared above according to the ratio of 1:100, and use the method of root irrigation to pour 50ml of the diluted bacterial liquid in each pot, once every 3 weeks, without adding bacteria The group poured the same dose of dead bacteria, and the concentration o...
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