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A phosphate-solubilizing bacterium strain, a compound bacterial agent including the same and a DEHP degrading bacterium and applications of the compound bacterial agent

A technology of phosphorus-dissolving bacteria and compound bacteria, which is applied in the field of microbial engineering, can solve problems affecting the quality and safety of agricultural products, and achieve the effects of promoting sustainable agricultural development, good synergy, and improving soil ecological fertility

Active Publication Date: 2018-09-18
JINAN UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

PAEs in contaminated soil are absorbed and accumulated by crops, seriously affecting the quality and safety of agricultural products

Method used

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  • A phosphate-solubilizing bacterium strain, a compound bacterial agent including the same and a DEHP degrading bacterium and applications of the compound bacterial agent
  • A phosphate-solubilizing bacterium strain, a compound bacterial agent including the same and a DEHP degrading bacterium and applications of the compound bacterial agent
  • A phosphate-solubilizing bacterium strain, a compound bacterial agent including the same and a DEHP degrading bacterium and applications of the compound bacterial agent

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] Isolation and screening of phosphate-soluble bacteria:

[0042] (1) Sample collection: Collect the activated sludge from the sewage treatment plant in a 50mL sterile centrifuge tube and place it in a clean ice box. Weigh 1g of activated sludge and dissolve it in a 50mL centrifuge tube with 40mL of sterile water, vibrate into a uniform (130rpm) in a shaker, and use 0.9% sterile saline to dilute the sample gradient to make 10 -1 ~10 -5 solution.

[0043] (2) Medium:

[0044] Monkina (PVK) liquid medium: Ca 3 (PO 4 ) 2 5.0g, sucrose 10.0g, (NH 4 ) 2 SO 4 0.5g, NaCl0.1g, MgSO 4 ·7H 2 O 0.1g, KCl 0.2g, MnSO 4 0.03g, yeast extract 0.5g, distilled water 1000mL, pH 7.0~7.2. Add 15g of agar to PVK solid medium.

[0045] Phosphate growth (NBRIP) medium: glucose 10.0g, Ca 3 (PO 4 ) 2 5.0g, MgCl 2 ·6H 2 O5.0g, MgSO 4 ·7H 2 O 0.25g, KCl 0.2g, (NH 4 ) 2 SO 4 0.1g, distilled water 1000mL, pH 7.0.

[0046] LB liquid medium: yeast powder 5.0g, peptone 10.0g,...

Embodiment 2

[0049] Identification of Phosphorous Solubilizing Bacteria Strain YLYP29:

[0050] The basis and method for the detection of morphological characteristics, physiology and biochemistry is the bacterial identification procedure MS(i) / C005-C01. Morphological identification: the bacteria is Gram-negative bacteria. Under the scanning electron microscope, it is short rod-shaped, with an average length of 0.8-1.0 μm and an average width of 0.2-0.3 μm ( figure 1 ); cultured at 30°C for 48h on LB solid medium, the colonies were yellowish-white, moist, mucus-like, smooth and opaque, with neat edges ( figure 2 ); the physiological and biochemical identification test results of the bacterial strain YLYP29 are shown in Table 1, the optimal growth conditions of the bacterial strain are: temperature 25-35° C., pH 6.0-8.0.

[0051] Identification of physiological and biochemical characteristics: See Table 1 for the results of physiological and biochemical experiments.

[0052] Table 1 Phy...

Embodiment 3

[0059] Determination of Phosphorus Solubilizing Ability of Strain Pseudomonas sp.YLYP29:

[0060] (1) Bacterial suspension preparation: use an inoculation loop to pick a small amount of YLYP29 strains stored on the slant of the test tube in 50mL LB liquid medium, culture at 30°C and 130rpm for 10h, take 40mL of culture solution in a 50mL centrifuge tube , centrifuged at 4°C and 5000rpm for 3min, collected the cells, washed the cells with 0.9% sterile normal saline for 3 times, and prepared a bacterial density of 10 with normal saline. 8 cfu / mL bacterial suspension.

[0061] (2) Insoluble phosphate pretreatment: Weigh 20.0g of analytically pure aluminum phosphate, iron phosphate or calcium phosphate into a large beaker, dissolve it ultrasonically with 2L of ultrapure water, discard the upper layer of turbid solution every 30min, and keep the bottom of the beaker Dissolved aluminum phosphate, iron phosphate or calcium phosphate, repeated ultrasonic washing 2 to 3 times until th...

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Abstract

A phosphate-solubilizing bacterium strain, a compound bacterial agent including the same and a DEHP degrading bacterium and applications of the compound bacterial agent are disclosed. The strain is Pseudomonas sp.YLYP29 deposited in the China Center for Type Culture Collection in Wuhan University, Wuhan, China on June 28, 2017 with an accession number of CCTCC NO:M 2017391. The compound bacterialagent consists of the phosphate-solubilizing bacterium YLYP29 and the DEHP degrading bacterium XB, has good synergistic effects, high adaptive capacity, phosphate-solubilizing and DEHP degrading functions and a good application potential, and a culture method is simple and feasible. The compound bacterial agent can solve the problem of soil phosphorus deficiency, can remove DEHP pollutants in soil, improves soil nutrition states, improves soil ecological fertility and restores DEHP polluted soil, thus ensuring agricultural product quality and yield. The compound bacterial agent plays an important role in restoration of farmland soil along with production.

Description

technical field [0001] The invention belongs to the technical field of microbial engineering, and in particular relates to a phosphate-dissolving bacterium and its compound bacterial agent with DEHP degrading bacteria and its application. Background technique [0002] Phosphorus is one of the important fertilizers in the soil, which promotes the growth and development of plants. Among the different forms of phosphorus compounds, only soluble phosphorus can be absorbed and utilized by plants, but 95% of phosphorus in soil exists in a poorly soluble state. About 2 / 3 of the cultivated land in our country is in a state of phosphorus deficiency. In order to ensure the increase of agricultural production and income, a large amount of chemical fertilizers are used in agricultural production. Ca 2+ , Fe 3 +, Al 3+ Such cations are fixed to form insoluble phosphate, and induce soil trace element deficiency, nutrient imbalance, soil compaction and other problems. Some soluble pho...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20B09C1/10C12R1/38
CPCB09C1/10C12N1/205C12R2001/38
Inventor 莫测辉冯乃宪李彦文蔡全英赵海明李慧
Owner JINAN UNIVERSITY
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