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Providencia alcalifaciens and application thereof in degrading tetracycline and generating phytohormone

A Providencia and auxin technology is applied in the field of antibiotic-contaminated microbial remediation to achieve the effect of reducing residues

Active Publication Date: 2020-05-01
SOUTH CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Nowadays, the ability of microorganisms to secrete IAA is used as a main indicator of microorganisms to promote plant growth, but there are few reports in existing studies that have the ability to simultaneously degrade TC and secrete IAA

Method used

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  • Providencia alcalifaciens and application thereof in degrading tetracycline and generating phytohormone
  • Providencia alcalifaciens and application thereof in degrading tetracycline and generating phytohormone
  • Providencia alcalifaciens and application thereof in degrading tetracycline and generating phytohormone

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0045] Screening, isolation, purification and identification of Example 1 bacterial strain TC1

[0046] 1. Screening method for a tetracycline-degrading bacterial strain TC1

[0047] 1. Material preparation

[0048] Soil source for strain screening: It was taken from fertilized vegetable fields of farmhouses in Zuolong Village, Chaling County, Zhuzhou City, Hunan Province. The soil samples were sealed with sampling bags, and brought back to the laboratory at 4°C for storage for later use.

[0049] LB medium: peptone 10.0g, yeast extract powder 5.0g, NaCl 10.0g, pH7.0-7.2, distilled water to 1L; solid medium plus 18.0g agar powder. Sterilize at 121°C for 15 minutes.

[0050] Inorganic salt medium (MSM medium): 5mL phosphate buffer solution (KH 2 PO 4 8.5g·L -1 、K 2 HPO 4 ·H 2 O2 1.75g·L -1 、Na 2 HPO 4 12H 2 O 33.4g·L -1 , NH 4 Cl 5.0g L -1 ), 3.0mL 22.5g·L -1 MgSO 4 Solution (MgSO 4 ·7H 2 O 46.125g·L -1 ), 1.0mL 0.25g·L -1 FeCl 3 Solution (FeCl 3 ·6H 2...

Embodiment 2

[0074] Degradation of Tetracycline by Example 2 Bacterial Strain TC1 in Inorganic Salt Medium Containing Tetracycline and Peptone

[0075] Streak the strain TC1 and culture it on the LB plate at 30°C for 20 hours, pick a single colony and inoculate it in the LB liquid medium, 30°C, 150r min -1 After culturing in a shaker for 20 hours, inoculate at a ratio of 2% (v / v) to a TC content of 10 mg·L -1 containing 10g·L -1 Cultured in tryptone-based MSM medium, and samples were taken after 3, 5, and 9 d (days); no addition of strain TC952 was used as a control (CK). Sample liquid at 10000r·min -1 After centrifugation for 1 min, the supernatant was filtered with a 0.22 μm filter membrane, and the residual amount of TC was measured by HPLC after filtration (tetracycline determination method was the same as in Example 1).

[0076] The results showed that the control TC without adding bacteria was hydrolyzed, and after adding bacteria TC1, the degradation of TC was significantly accel...

Embodiment 3

[0077] Example 3 Qualitative Test of Auxin Secretion of Bacterial Strain TC1

[0078] Inoculate the strain TC1 drawn on the LB solid medium into the LB liquid medium, shake and culture at 125-150 rpm, 28-30°C for 18-24 hours, and then inoculate it into the King culture with 2% inoculation volume (V / V) In the basal medium (TC1 treatment group), 2% sterile water was added to the King's medium for treatment as a negative blank control (CK-), placed at 125 rpm, and shaken at 28° C. for 48 hours (each treatment was repeated three times). At room temperature, pipette 50ul of the treatment solution of each group onto the white color comparison plate, and add 50μL of the color comparison solution at the same time; observe the color change of each group after dark treatment for 15 minutes, if the color is pink, the reaction is positive, indicating that it can secrete and produce IAA, and the darker the color, the stronger the ability to secrete IAA; otherwise, it is negative ( Figure...

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Abstract

The invention discloses providencia alcalifaciens and application thereof in degrading tetracycline and generating phytohormone. The providencia alcalifaciens is named as Providencia alcalifaciens TC1, the preservation number is GDMCC NO:60787, and the strain is preserved in the Guangdong Microbiological Culture Collection Center on Floor 5, Building 59, No.100 Courtyard, Middle Xianlie Road, Guangzhou on September 25, 2019. The Providencia alcalifaciens TC1 disclosed by the invention is capable of degrading tetracycline in a co-metabolism mode and secreting auxin or analogs, therefore, the Providencia alcalifaciens TC1 can be applied to degradation of medium / low-concentration tetracycline in soil and water bodies to remove or reduce residues of tetracycline in the environment, and can bealso used to generate phytohormone to promote growth of plants.

Description

technical field [0001] The invention belongs to the technical field of antibiotic-contaminated microorganism restoration, and in particular relates to a strain of Alcaligenes providencia and its application in degrading tetracycline and producing plant auxin. Background technique [0002] At present, antibiotics are increasingly used in veterinary and medical fields. Tetracycline (TC), as a broad-spectrum antibiotic, is widely used in the poultry industry because of its good inhibitory effect on Gram, some rickettsia, viral viruses and protozoa. When TC is used in humans or animals, part of the unused tetracycline or biological metabolites will be excreted with animal urine and feces, and the application of tetracycline-containing poultry and livestock manure as organic fertilizers on farmland will pollute Soil systems can even contaminate groundwater systems. In addition, tetracycline also enters soil and water through pharmaceutical wastewater discharge and discarded exp...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12P17/10A01N63/20A01P21/00B09C1/10C02F3/34C12R1/01C02F101/38
CPCA01N63/00B09C1/10C02F3/34C02F2101/38C12N1/20C12P17/10C12N1/205C12R2001/01
Inventor 龚贝妮谭泽文李永涛徐会娟邹艺璇邓翠文
Owner SOUTH CHINA AGRI UNIV
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