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A high-efficiency pesticide-degrading Arthrobacter and its screening and application

An Arthrobacter and strain technology, applied in the field of microorganisms, can solve the problems of interfering with the development of sperm and testicles of adult mice, destroying the development of mammalian uterus, increasing the stillbirth rate, etc., to achieve the effect of promoting sustainable development, improving quality and safety, and reducing pollution

Active Publication Date: 2021-05-18
BAOJI UNIV OF ARTS & SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

(2) Reproductive toxicity: Carbendazim is an endocrine disruptor that can disrupt the development of mammalian uterus and interfere with the development of sperm and testis in adult mice
(3) Embryo toxicity: Male mice fed carbendazim for 60 days were mated with normal female mice, and it was found that although the pregnancy rate and average implantation number of female mice were not significantly affected, the stillbirth rate increased
Since carbendazim is absorbed by sediment in the water, toxicity is usually not high, but organisms that depend on sediment for survival are likely to be affected by high doses of carbendazim

Method used

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  • A high-efficiency pesticide-degrading Arthrobacter and its screening and application
  • A high-efficiency pesticide-degrading Arthrobacter and its screening and application
  • A high-efficiency pesticide-degrading Arthrobacter and its screening and application

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Screening, isolation and purification of embodiment 1 bacterial strain djl-7H

[0029] (1) Sample collection: The test soil was collected from the greenhouse soil where carbendazim was continuously applied in the College of Horticulture, Northwest Agriculture and Forestry University. The target strain was isolated and purified from the collected test soil.

[0030] (2) Medium:

[0031] a) LB liquid medium: NaCl 10.0g, peptone 10.0g, yeast extract powder 5.0g, add distilled water 1000mL, adjust the pH value to 7.0.

[0032] b) Basal salt medium: NaCl 1.0g, K 2 HPO 4 1.5g, KH 2 PO 4 0.5g, (NH 4 ) 2 SO 4 2.0g, MgSO 4 ·7H 2 O0.2g, distilled water 1000mL, pH=7.0.

[0033] c) LB solid medium: same as LB liquid medium, with 1%-2% agar added.

[0034] d) Medium for enrichment (degradation) test: Same as basic salt medium, with carbendazim added as needed.

[0035] e) Separation and purification medium: the same as the enrichment (degradation) test medium, adding 1% ...

Embodiment 2

[0043] Identification test of embodiment 2 bacterial strain djl-7H

[0044] (1) Morphological features

[0045] The colony of the djl-7H strain is round, milky white, translucent, raised, with complete edges, smooth surface, moist, sticky and difficult to pick, and the colony size is (0.3-0.5) cm × (0.3-0.5) cm (such as figure 1 ).

[0046] Morphological observation: The morphology of the tested strain was observed by optical microscope and scanning electron microscope. The morphological characteristics are: rod-shaped with a small amount of flagella, the size of the bacteria is 0.2-0.5 μm, 1.0-3.0 μm, and it grows relatively fast on the medium. fast. (Such as figure 2 ).

[0047] Physiological and biochemical indicators: refer to "Bergey's Manual of Determinative Bacteriology" and "Common Bacterial System Identification Manual". Specific determination indexes include: standard catalase, starch hydrolysis, indole test, Gram staining, methyl red, V-P and 3-ketolactose in...

Embodiment 3

[0062] The degradation test of embodiment 3 joint bacillus (Arthrobacter sp.) djl-7H to carbendazim

[0063] (1) Preparation of seed solution

[0064] Pick a ring of bacteria, inoculate into a Erlenmeyer flask filled with LB liquid medium, shake and culture at 30°C for about 24 hours; use sterilized LB liquid medium to adjust its OD 600 are all 1.0, pipette 5mL of bacterial solution into a 10mL centrifuge tube, centrifuge at 4°C and 4000r / min for 5min, and collect the bacterial cells; wash twice with sterilized basic salt medium, then use an equal volume of Suspend the basal salt medium and set aside.

[0065] (2) Degradation test of carbendazim

[0066] 100 mg / L carbendazim was added to the medium for degradation test, and 5% inoculum was inserted into the seed solution of carbendazim-degrading bacteria djl-7H, and cultured at 25°C and 150r / min with shaking. Sampling was carried out at regular intervals of 6 hours until 72 hours of cultivation, and the carbendazim and meta...

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Abstract

The invention discloses an Arthrobacter that efficiently degrades pesticides and its screening and application. The strain has been preserved in the China Center for Type Culture Collection on May 30, 2018, with a preservation number of CCTCC NO: M 2018328. The bacterial strain of the invention can effectively degrade carbendazim, which lays a foundation for solving the problem of carbendazim pesticide residues, and also provides materials for the research and development of efficient carbendazim-degrading enzymes.

Description

technical field [0001] The invention belongs to the technical field of microorganisms, relates to a new Arthrobacterium (Agrobacterium), in particular to an Arthrobacter sp. degradation applications. Background technique [0002] Carbendazim (carbendazim), also known as cotton wilting, befenti, benzimidazole No. 44, English trade name MBC, relative molecular mass is 191.2, molecular formula is: C 9 h 9 N 3 0 2 , the chemical structural formula is as follows: [0003] [0004] Carbendazim is a broad-spectrum fungicide. Since 2011, its annual usage has been about 40,000 tons. In 2012, its registered products reached 857. The dosage forms include technical medicine, powder, suspension concentrate, wettable powder, and suspoemulsion , Seed coating agent and so on nearly ten kinds. In my country, carbendazim is widely used in the prevention and control of diseases of wheat, fruit trees, tea leaves, vegetables, etc., and is also used for post-harvest preservation of fruit...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20C12N1/02C02F3/34B09C1/10C12R1/06C02F101/38
CPCB09C1/10C02F3/34C02F2101/38C12N1/02C12N1/20C12N1/205C12R2001/06
Inventor 雷霁范拴喜周进宏赵倩赵钺
Owner BAOJI UNIV OF ARTS & SCI