A method for screening high-efficiency nitrogen-fixing and phosphorus-dissolving bacteria

A technology of phosphorus-dissolving bacteria and nitrogen fixation, applied in the field of environmental microorganisms, can solve the problems of low screening efficiency, low screening bacteria pertinence, long enrichment time, etc., and achieve the effect of shortening the screening time and the strain enrichment time.

Active Publication Date: 2022-04-08
安徽建工交通航务集团有限公司 +1
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  • Summary
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  • Description
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  • Application Information

AI Technical Summary

Problems solved by technology

The enrichment time of this method is longer, and the bacteria screening is less targeted, which greatly reduces the efficiency of bacteria screening

Method used

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specific Embodiment approach

[0015] The method for screening high-efficiency nitrogen-fixing and phosphorus-dissolving bacteria of the present invention, its preferred embodiment is:

[0016] Include the following steps:

[0017] (a) first mix the soil with water and then culture with shaking, centrifuge and discard the supernatant;

[0018] (b) Adding a phosphate-dissolving medium containing an additional carbon source to the precipitate in proportion for culturing;

[0019] (c) centrifuging the culture solution obtained in step (b) and discarding the supernatant, adding a nitrogen-fixing medium without additional carbon source in proportion for culturing;

[0020] (d) centrifuging the culture solution obtained in step (c) and discarding the supernatant, adding a phosphate-dissolving medium without additional carbon source in proportion for culturing;

[0021] (e) alternately cultivating multiple times according to the method of steps (c) and (d);

[0022] (f) The target strain is screened out by usin...

specific Embodiment

[0041] Step 1: Pretreatment of Soil Samples

[0042] Take 10g of fresh soil into a 250m Erlenmeyer flask, add 100mL of sterile water, shake and cultivate at 150rpm at 30°C for 24 hours, then centrifuge at 4000rpm, and discard the supernatant by decantation to obtain a precipitate.

[0043] Step two:

[0044] Add 100 mL of phosphate-dissolving medium containing an external carbon source into the Erlenmeyer flask containing the precipitate, and shake at a constant temperature of 150 rpm at 30°C for 24 hours. After cultivation, water samples were taken to detect the total phosphorus content.

[0045] Among them, the phosphate-dissolving medium with an external carbon source includes the following components:

[0046] 10g / L Glucose, 5g / L CaPO 4 , 0.1g / L (NH 4 ) 2 SO 4 , 0.2g / L KCl, 0.25g / L MgSO 4 ·7H 2 O, 5g / L MgCl 2 ·6H 2 O, 1mL / L trace element one solution, pH 7.0-7.5, sterilized at 121°C for 30min.

[0047] Described trace element-solution comprises following compositi...

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Abstract

The invention discloses a method for screening high-efficiency nitrogen-fixing and phosphorus-dissolving bacteria. Firstly, the soil is mixed with water and cultured by shaking, and the supernatant is discarded after centrifugation; a phosphorus-dissolving medium containing an external carbon source is added to the precipitation in proportion to carry out cultivation Discard the supernatant after the obtained culture solution is centrifuged, add nitrogen-fixing medium without additional carbon source in proportion, and cultivate; discard the supernatant after the obtained culture solution is centrifuged, and add phosphorus-dissolving medium without external carbon source in proportion , and cultured; after repeated cultures alternately, the target strain was screened out using phosphorus-dissolving and nitrogen-fixing medium. Screening is carried out by using nitrogen-fixing medium and phosphate-dissolving medium for alternate culture, and then the acclimatization medium is diluted and spread to screen out single strains. The screening cycle is greatly shortened. It has the advantages of strong selectivity, small workload, and can greatly shorten the screening cycle. It is an efficient and rapid screening method for nitrogen-fixing and phosphorus-dissolving bacteria.

Description

technical field [0001] The invention relates to an environmental microbial technology, in particular to a method for screening high-efficiency nitrogen-fixing and phosphorus-dissolving bacteria. Background technique [0002] The lack of nitrogen and phosphorus elements in the soil is an important factor affecting the biomass of food crops or other plants. Generally, the nitrogen and phosphorus content in the soil is increased by applying a large amount of nitrogen fertilizer and phosphorus fertilizer. However, the utilization rate of this kind of chemical fertilizer is low. If the nitrogen and phosphorus compounds that are not used by plants in time cannot be absorbed by the soil colloid, they will be transferred to the depth of the soil with the infiltrating soil water, causing pollution, which can lead to the pollution of rivers and lakes. Eutrophication etc. However, direct screening of microorganisms with efficient nitrogen fixation and phosphorus dissolution from the s...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/02
CPCC12N1/02
Inventor 邵佳孙庆业黄国平邱峰
Owner 安徽建工交通航务集团有限公司
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