Composite nitrogen-fixing microbial agent as well as preparation method and application thereof
A technology of nitrogen-fixing microorganisms and nitrogen-fixing bacteria, applied in the field of microbial applications, can solve problems such as imperfect research and testing of bacteria, short duration of drug effects, and lack of cooperation between bacteria and bacteria, so as to increase the soluble sugar content of plants and improve the diversity of soil microorganisms , increasing the effect of diversity
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Embodiment 1
[0049] The preparation of embodiment 1 composite nitrogen-fixing microbial bacterial agent
[0050] The preparation of composite nitrogen-fixing microbial bacterial agent mainly comprises the following steps:
[0051] 1. Single plant culture of microorganisms
[0052] (1) Separate culture of nitrogen-fixing bacteria
[0053] Obtain nitrogen-fixing bacteria powder or bacterial liquid or test tube slope from commercial sources, put the nitrogen-fixing bacteria sample in 0.3mL nitrogen-fixing bacteria fermentation broth, obtain the suspension of nitrogen-fixing bacteria, and use the recovered nitrogen-fixing bacteria suspension or colony as seeds Add the nitrogen-fixing bacteria fermentation liquid into the nitrogen-fixing bacteria fermentation broth, and inoculate the nitrogen-fixing bacteria strains in the fermentation broth with pH=7.2 according to the ratio of adding 0.3mL seed liquid to every 100mL fermentation broth. Ferment for 24-48 hours until the effective number of v...
Embodiment 2
[0077]The performance detection of embodiment 2 composite nitrogen-fixing microbial bacterial agents
[0078] In this embodiment, performance testing of the composite nitrogen-fixing microbial bacterial agent prepared in Example 1 will be carried out through the following four aspects.
[0079] 1. Hemolytic reaction of microbial agents
[0080] The mixed fermentation broth prepared in Example 1 was inoculated on a blood agar plate, and cultured at 37° C. for 24 hours. It was observed that no hemolytic zone was produced, indicating that the composite nitrogen-fixing microbial bacterial agent prepared by the present invention can be used as an agricultural microbial bacterial agent.
[0081] The formula of the blood agar plate in the test is as follows: 10g of peptone, 10g of beef extract powder, 5g of sodium chloride, 15g of agar, 1L of distilled water, pH=7.5, autoclave and cool to 60°C, add sterile defibrated sheep blood, rotate and fully pour the plate , blood agar layer th...
Embodiment 3
[0095] Embodiment 3 Composite nitrogen-fixing microbial agent and plant interaction relationship test
[0096] In this example, the interaction between the compound nitrogen-fixing microbial agent and the plant will be explored from the two aspects of microbial colonization in the plant rhizosphere environment and the impact on plant tissue.
[0097] 1. The colonization of microorganisms in the plant rhizosphere environment
[0098] The potted crops are planted in the incubator, and the potting soil is humus soil, which comes from Changbai Mountain in Jilin Province. The cultivation pot used in the experiment is 21cm*21cm. The physical and chemical properties of the soil are: the average particle size in dry state is 223μm, the wet state particle size is 66.5μm, and the particle volume is 4.64*105cm 3 , The density of dry humus is about 2.47g / cm3. The mass fraction of organic matter is 29.13%, the solid content rate is 89.50%, and the planting plants are peanuts. After soak...
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Abstract
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