Culture method of efficient nitrogen-fixing bacteria in water body
A cultivation method and technology for nitrogen-fixing bacteria, which can be used in microorganism-based methods, chemical instruments and methods, biochemical equipment and methods, etc., and can solve the problems of long nitrogen-fixing cycle and unsatisfactory nitrogen-fixing effect.
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Embodiment 1
[0037] 1) Inoculate Bacillus megaterium with an inoculum size of 3%-5% in MS medium (containing nitrosoethylurea, the total content is 1.8% by weight) and carry out preliminary culture (cultivation temperature is 25°C, culture time for 8 days, the relative humidity is 50%);
[0038] 2) Place the culture medium under gamma rays for two-step culture (the culture temperature is 25°C, the culture time is 20h, the relative humidity is 50%, and the energy of the gamma rays is 1 Mev);
[0039] 3) Place the culture medium for freezing treatment (the treatment temperature is -12°C, and the treatment time is 9h), and then the temperature is raised to 25°C;
[0040] 4) Select the mixed flora in the culture medium and use the plate marking method (A, B, C and D four areas connected in sequence along the plate design, the area of the four areas is D>C>B>A); select D area The bacteria are high-efficiency nitrogen-fixing bacteria in water.
Embodiment 2
[0042] 1) Inoculate Bacillus megaterium with 3% inoculum in MS medium (containing nitrosoethylurea and nitrosomethylurea, the total content is 1.2% by weight) and carry out preliminary culture (cultivation temperature is 15 ℃, the incubation time is 10 days, and the relative humidity is 60%);
[0043] 2) Place the culture medium under gamma rays for two-step culture (the culture temperature is 15°C, the culture time is 24h, the relative humidity is 60%, and the energy of the gamma rays is 1.5 Mev);
[0044] 3) Place the culture medium for freezing treatment (the treatment temperature is -15°C, and the treatment time is 8h), and then the temperature is raised to 15°C;
[0045] 4) Select the mixed flora in the culture medium and use the plate marking method (A, B, C and D four areas connected in sequence along the plate design, the area of the four areas is D>C>B>A); select D area The bacteria are high-efficiency nitrogen-fixing bacteria in water.
Embodiment 3
[0047] 1) Inoculate Bacillus megaterium with 5% inoculum in 1 / 2MS medium (containing nitrosoethylurea and / or nitrosomethylurea, the total content is 2% by weight) for preliminary culture ( The culture temperature is 35°C, the culture time is 5 days, and the relative humidity is 40%);
[0048] 2) Place the culture medium under gamma rays for two-step culture (the culture temperature is 35°C, the culture time is 15h, the relative humidity is 40%, and the energy of the gamma rays is 0.8 Mev);
[0049] 3) Place the culture medium for freezing treatment (the treatment temperature is -10°C, and the treatment time is 10h), and then the temperature is raised to 35°C;
[0050] 4) Select the mixed flora in the culture medium and use the plate marking method (A, B, C and D four areas connected in sequence along the plate design, the area of the four areas is D>C>B>A); select D area The bacteria are high-efficiency nitrogen-fixing bacteria in water.
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