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Screening and identifying method and application of jinhua strain capable of degrading cypermethrin

A technology of golden flower fungus and strain, which is applied in the field of screening and identification of cypermethrin degrading "Golden flower fungus", which can solve the problems of high detection rate and over-standard rate, residual accumulation, slow natural degradation rate, etc.

Inactive Publication Date: 2015-02-04
SICHUAN AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Due to its strong hydrophobicity and light and heat stability, its natural degradation rate is slow, and its half-life in the environment is as long as 94.2 d-1103 d, which can easily lead to its residual accumulation in the environment. High detection rate and exceeding standard rate often appear in the product

Method used

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  • Screening and identifying method and application of jinhua strain capable of degrading cypermethrin
  • Screening and identifying method and application of jinhua strain capable of degrading cypermethrin
  • Screening and identifying method and application of jinhua strain capable of degrading cypermethrin

Examples

Experimental program
Comparison scheme
Effect test

example 1

[0026] Example 1: Determine the relationship between the growth of cypermethrin-degrading strain ET1 and the degradation of cypermethrin.

[0027] 1) Strain activation: Pick the cypermethrin-degrading strain ET1 into the PD liquid medium, 30°C, 180r / min shaker shaking culture for 4 days; then continuously streak on the PDA plate, pick a single colony and culture twice (30°C ) 4d, pick a single colony and culture it on a PDA slope at 30°C for 4d; 8 cells / mL to make a seed solution.

[0028]Determination of the growth curve: inoculate the seed solution of the degrading strain ET1 in the PD liquid medium containing 50 mg / L cypermethrin according to the inoculation amount of 5%, aliquot into 30 mL / 250 mL Erlenmeyer flasks, and culture at 30 °C and 180 r / min for 8 days , The sampling time is to sample 1 bottle every 24 hours, filter the sample to obtain mycelium balls, dry them at 80°C to constant weight, and use the dry weight of the bacteria to represent its biomass.

[0029] D...

example 2

[0032] Example 2: Degradation characteristics of cypermethrin by degrading strain ET1.

[0033] 1) Effect of culture temperature on the degradation rate of cypermethrin by strain ET1 According to the inoculum size of 5.0% (v / v), the strain seed solution (6.0×10 8 permL) inoculated in PD liquid medium (pH6.0) with a concentration of cypermethrin of 50 mg / L, distributed into 30 mL / 250 mL Erlenmeyer flasks, at different temperatures (25°C, 30°C, 35°C) Shake culture at 180 r / min, and take samples at intervals of 24 hours. Measure the degradation rate of cypermethrin with the method described in [0023].

[0034] 2) Effect of pH on the degradation rate of cypermethrin by strain ET1 According to the inoculum size of 5.0% (v / v), the strain seed solution (6.0×10 8 permL) were inoculated in PD liquid medium with cypermethrin concentration of 50 mg / L at different pH (5.0, 6.0, 7.0), divided into 30 mL / 250 mL Erlenmeyer flasks, cultured with shaking at 30 °C and 180 r / min , Sampling ...

example 3

[0037] Example 3: Degradation effect of cypermethrin-degrading strain ET1 on its degradation intermediate product 3-phenoxybenzoic acid.

[0038] The seed solution of the degraded strain was inoculated in 50 mg / L 3-phenoxybenzoic acid PD liquid medium according to the inoculation amount of 5%, and divided into 30 mL / 250 mL Erlenmeyer flasks, and the inoculation of 5% sterile water was set as the blank control. Shake culture at 30°C and 180r / min for 7 days, adopt the method of full sampling, that is, take the whole bottle of culture solution as an analysis sample, and measure its degradation rate of 3-phenoxybenzoic acid.

[0039] Add 30mL of acetonitrile to the whole bottle of culture solution, ultrasonic (40kHz, 300W) assisted extraction for 30min, pipette 2.0mL of extract solution and dilute to 10mL with acetonitrile, mix well, centrifuge at 12000r / min for 10min, and filter the supernatant with organic phase membrane (0.45 μm) filter, discard the initial filtrate, and take t...

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Abstract

The invention provides a bacterial strain ET1 with higher degradation ability for cypermethrin, which is screened from fuzhuan brick tea, belonging to the biological technical field. A registry number of an ITS (Internal Transcribed Spacers) gene sequence of the bacterial strain in a GenBank database is KF317836; the bacterial strain is identified as eurotium cristatum through morphological and ITS sequence analysis, and is a nontoxic and beneficial bacterium and preserved in China General Microbiological Culture Collection Center (CGMCC) on October 8, 2013 with a preservation number of CGMCC No.8293. The bacterial strain further has a certain degradation effect for other pyrethroid pesticides, and can degrade toxic intermediate products (3-phenoxybenzoicacid) of the pyrethroid pesticides. The bacterial strain not only can be used for fuzhuan brick tea and tea leaves polluted by the pyrethroid pesticides, but also can be used for bioremediation or biological purification of soil and wastewater to protect an ecological environment, so that safety of agricultural products is ensured.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to the screening and identification of a strain of "Golden Flower Fungus" degrading cypermethrin and its application. Background technique [0002] Cypermethrin (CY) is one of the most commonly used varieties of pyrethroid pesticides, and it is widely used in the control of pests such as tea trees, fruits and vegetables, humans and animals. Due to its strong hydrophobicity and light and heat stability, its natural degradation rate is slow, and its half-life in the environment is as long as 94.2 d-1103 d, which easily leads to its residual accumulation in the environment. Higher detection rate and exceedance rate often appear in the product. Cypermethrin is highly toxic to fish and other aquatic animals. It can also accumulate in animals and plants, has acute toxicity to the nervous system, and can damage the development of the male reproductive system. Long-term exposure may also cause...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/14A62D3/02A23F3/10B09C1/10C02F3/34C12R1/645A62D101/04A62D101/28C02F101/38
Inventor 刘书亮李金永文涛李建龙邹立扣
Owner SICHUAN AGRI UNIV
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