Multifunctional engineering strain for degrading chlorpyrifos and construction method thereof

An engineering strain, chlorpyrifos technology, applied in the field of biodegradation of chemical pesticides, can solve pollution and other problems, achieve the effect of promoting plant growth, increasing the function of reducing pesticide residues, and good repair effect

Inactive Publication Date: 2011-05-25
BIOTECH CENT OF SHANDONG ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] The purpose of the present invention is to provide a Vietnamese Burkholderia vietnamiensis multifunctional engineering strain (Burkholderia vietnamiensis) P418

Method used

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  • Multifunctional engineering strain for degrading chlorpyrifos and construction method thereof
  • Multifunctional engineering strain for degrading chlorpyrifos and construction method thereof
  • Multifunctional engineering strain for degrading chlorpyrifos and construction method thereof

Examples

Experimental program
Comparison scheme
Effect test

Example Embodiment

[0044] Example 1: Construction of a multifunctional engineered strain of chlorpyrifos

[0045] 1) Construction of recombinant plasmid pUT-M

[0046] Design primers based on organophosphate degrading enzyme genes:

[0047] mf: 5’GATCGCGGCCGCAGGAGGTTGATATGCCCCTGAAGAAC-3’

[0048] mr: 5’-GATAGCGGCCGCCTTGGGGTTGACGACCG-3’

[0049] Introduce Not I restriction site and SD sequence at both ends of the gene, and use pGM-T-M as template for PCR amplification. The reaction conditions are: pre-denaturation at 94°C for 3 minutes, denaturation at 94°C for 30 minutes, annealing at 55°C for 50 minutes, extension at 72°C for 1 minute, 30 cycles in total, and finally extension at 72°C for 10 minutes.

[0050] The organophosphate degrading enzyme gene and plasmid pUT-Km1 were cut with Not I, and then ligated, transformed into E.coli S17-1 (λ-pir), and the transformants were detected by PCR using the above primers. The reaction conditions were the same as above.

[0051] Detect the positive clones by PCR, e...

Example Embodiment

[0065] Example 2: Biological characteristics of chlorpyrifos multifunctional engineering strain P418-M

[0066] (1) Determination of the growth curve of the multifunctional engineering strain P418-M and the original strain P418

[0067] The growth rate of the two strains was measured in P418 liquid medium. The results show that( Figure 5 ), the engineered strain has a foreign gene inserted into the chromosome, but its growth curve is not significantly different from that of the original strain P418, indicating that the introduction of foreign genes only increases the function of the original strain without affecting its growth. This shows that the medium conditions of the original strain can be fully used for the cultivation of engineered strains.

[0068] (2) Comparison of the ability of dissolving phosphorus, potassium and nitrogen between the multifunctional engineering strain P418-M and the original strain P418

[0069] The following methods were used to determine the ability of...

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Abstract

The invention discloses a multifunctional engineering strain for degrading chlorpyrifos and a construction method thereof, and relates to a burkholderia vietnamiensis multifunctional engineering strain P418-M. The collection number of the strain is CGMCC No. 4166. The construction method for the burkholderia vietnamiensis multifunctional engineering strain comprises the following steps that: organic phosphorus degradation enzyme genes from acinetobacter are integrated onto chromosomes of burkholderia vietnamiensis by suicide plasmid through Tn5 transposition by using the burkholderia vietnamiensis, so the function of degrading pesticide residues is increased on the basis of the capacity of removing phosphorus and potassium, fixing nitrogen and promoting plant growth and rhizosphere colonization of a multifunctional microorganism to realize multiple functions of the burkholderia vietnamiensis.

Description

technical field [0001] The invention belongs to the technical field of biodegradation of chemical pesticides, and relates to a multifunctional engineering strain with high-efficiency degradation ability to organophosphorus pesticide chlorpyrifos, nitrogen fixation, phosphorus decomposing, potassium decomposing and bio-control functions and a construction method thereof. Background technique [0002] Chlorpyrifos (chlorpyrifos), also known as Lesben, chlorpyrifos, etc., the chemical name is O, O-diethyl-O-(3,5,6-trichloro-2-pyridyl) phosphorothioate, Developed by Dow Chemical Company in the United States in 1965, it has the characteristics of high efficiency, broad spectrum and high efficiency, and is widely used in the control of field crops, fruits and vegetables, flowers and household pests. [0003] Since my country banned highly toxic organophosphorus pesticides on January 1, 2007, chlorpyrifos quickly took the leading position as a low-toxicity and high-efficiency organ...

Claims

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Application Information

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IPC IPC(8): C12N1/21B09C1/10A01N63/02C05F11/08
Inventor 张新建李红梅李纪顺黄玉杰任艳魏艳丽扈进冬杨合同
Owner BIOTECH CENT OF SHANDONG ACAD OF SCI
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