High-efficiency phosphate-resolving Pseudomonas and microbial agent and application of Pseudomonas

A microbial agent, Pseudomonas technology, applied in the field of microorganisms, can solve the problems that crops cannot be absorbed and utilized, phosphorus fertilizer utilization rate is only, phosphorus leaching loss, volatilization, etc., to increase crop yield, reduce dosage, and improve soil fertility Effect

Pending Publication Date: 2020-01-17
HENAN UNIVERSITY OF TECHNOLOGY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, after the phosphorus fertilizer enters the soil, most of the phosphorus will be leached, volatilized or mixed with Ca in the soil. + , Fe 3+ 、Al 3+ The combination of plasma forms insoluble phosphate, which cannot be absorbed and utilized by crops, resulting in only about 20% of the utilization rate of phosphate fertilizer

Method used

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  • High-efficiency phosphate-resolving Pseudomonas and microbial agent and application of Pseudomonas
  • High-efficiency phosphate-resolving Pseudomonas and microbial agent and application of Pseudomonas

Examples

Experimental program
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Effect test

Embodiment 1

[0016] Example 1: Screening of Phosphorus Solubilizing Bacteria and Determination of Phosphorus Solubilizing Ability

[0017] A phosphate-solubilizing bacterium was isolated and screened from the lake sediment sample by a ten-fold gradient dilution method, and the applicant named it 1416X3. The bacterial colony was round, yellow, rod-shaped, and Gram staining was negative. 16S rRNA gene sequencing identified the bacterium 1416X3 as Pseudomonas ( Pseudomonas sp.).

[0018] The strain 1416X3 was inoculated into LB liquid medium, and cultured with shaking at 28°C and 150 rpm for 24 h. Inoculate 2 mL of seed liquid into 250 mL Erlenmeyer flasks filled with 100 mL of inorganic phosphorus liquid medium, and use the medium without any bacteria as the blank control. Each treatment was repeated three times, and cultured with shaking at 28°C and 150 rpm for 5 Every 24 h, take the culture solution, centrifuge at 10000 rpm for 10 min, take the supernatant, and measure its soluble phosp...

Embodiment 2

[0021] Embodiment 2: the fermentation method of phosphate solubilizing bacteria

[0022] (1) Expanded cultivation of seeds: Take 1-2 lawns of Pseudomonas annulus 1416X3 from the slope, inoculate them in sterilized LB liquid medium for shaking culture; the culture conditions are temperature 25-30 ℃, rotation speed 150- 200 rpm, 500 mL Erlenmeyer flask with a liquid volume of 150-250 mL and a culture time of 24-36 h. The composition of the LB medium is: yeast extract 5 g / L, tryptone 10 g / L, NaCl 10 g / L, pH 7.0-7.2, solid medium plus 15 g agar, sterilized at 121 °C for 20 min.

[0023] (2) Fermentation culture: put the above seed liquid into the inorganic phosphorus fermentation medium with 1-5% inoculation amount (w / w), temperature 25-30°C, rotation speed 150-200 rpm, fermentation time 24-36 h ; The composition of the inorganic phosphorus medium is: glucose 15 g / L, MgCl 2 ·6H 2 O 5 g / L, MgSO 4 ·7H 2 O 0.25 g / L, KCl 0.2 g / L, (NH 4 ) 2 SO 4 0.2 g / L, Ca 3 (PO 4 ) 2 5g / ...

Embodiment 3

[0025] Embodiment 3: the influence of different culture temperature and initial pH value on Pseudomonas 1416X3

[0026] Pseudomonas 1416X3 was inoculated into LB liquid medium, and cultured with shaking at 28°C and 150 rpm for 24 h. Take 2 mL of cultured Pseudomonas 1416X3 seed liquid, inoculate it in a 250 mL Erlenmeyer flask containing 100 mL of inorganic phosphorus liquid medium, use the uninoculated culture liquid as a control, and inoculate them at 20°C, 28°C, and 37°C, respectively. Under the conditions of ℃ and 40℃, shake culture at 150 rpm for 5 days, repeat each treatment 3 times, take the fermentation broth every 24 h, centrifuge at 10,000 rpm for 10 min, take the supernatant, and use the molybdenum antimony anti-colorimetric method to determine the content of available phosphorus. content, the result is figure 1 shown. the result shows( figure 1 ), Pseudomonas 1416X3 had the best phosphorus-dissolving effect at 28°C. After optimizing the culture temperature, its ...

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Abstract

The invention belongs to the technical field of microorganisms, and specifically relates to a strain of high-efficiency phosphate-resolving Pseudomonas and a microbial agent and application of the Pseudomonas. The strain was deposited on April 11, 2019 at General Microbiological Center of China Microbiological Culture Collection Management Committee, located at No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing, and the deposited number is CGMCC No. 17557. The bacteria can transform insoluble inorganic phosphorus into soluble available phosphorus with high efficiency, and the amountof dissolved phosphorus can reach 793 mg/L when calcium phosphate is used as the sole phosphorus source. The bacteria can be prepared into a phosphate-resolving microbial agent, which can be widely used in agricultural production, the content of the available phosphorus in soil can be effectively increased, the effect of saving fertilizer and increasing production can be achieved, and the microbial agent has good social, economic and environmental benefits.

Description

technical field [0001] The invention belongs to the technical field of microorganisms, and in particular relates to a high-efficiency Pseudomonas phospholytica and its microbial agent and application. Background technique [0002] Phosphorus is one of the essential nutrients for plant growth and participates in most of the metabolic processes in plants. However, phosphorus in the environment usually exists in the form of calcium phosphate salts, which cannot be directly absorbed and utilized by plants, resulting in severe phosphorus deficiency in most cultivated land. As phosphorus is a non-renewable mineral resource, insufficient phosphorus content in soil has become an important factor restricting crop yield. Therefore, in agricultural production, the above-mentioned problems are usually solved by applying phosphorus fertilizers. However, after the phosphorus fertilizer enters the soil, most of the phosphorus will be leached, volatilized or mixed with Ca in the soil. + ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20A01N63/27A01P21/00C05F11/08C05G3/00C05G3/80C12R1/38
CPCC12N1/20A01N63/00C05F11/08C05G3/00C12R2001/38C12N1/205
Inventor 周佳屈建航田海龙王喜凤李海峰屈凌波王林风
Owner HENAN UNIVERSITY OF TECHNOLOGY
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