CN105567597A - High-efficiency Atrazine degrading bacteria and application and screening method thereof - Google Patents

High-efficiency Atrazine degrading bacteria and application and screening method thereof Download PDF

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CN105567597A
CN105567597A CN201610026144.3A CN201610026144A CN105567597A CN 105567597 A CN105567597 A CN 105567597A CN 201610026144 A CN201610026144 A CN 201610026144A CN 105567597 A CN105567597 A CN 105567597A
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atrazine
screening method
efficient
atrazine degradation
bacterium
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CN105567597B (en
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马放
赵昕悦
王立
杨基先
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Yixing Environmental Protection Industry Co.,Ltd.
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HIT YIXING ACADEMY OF ENVIRONMENTAL PROTECTION
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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
    • C12R2001/00—Microorganisms ; Processes using microorganisms
    • C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
    • C12R2001/06—Arthrobacter
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
    • C12N1/20—Bacteria; Culture media therefor
    • C12N1/205—Bacterial isolates
    • C—CHEMISTRY; METALLURGY
    • C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02F3/00—Biological treatment of water, waste water, or sewage
    • C02F3/34—Biological treatment of water, waste water, or sewage characterised by the microorganisms used
    • C—CHEMISTRY; METALLURGY
    • C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02F2101/00—Nature of the contaminant
    • C02F2101/30—Organic compounds
    • C02F2101/38—Organic compounds containing nitrogen

Abstract

The invention relates to high-efficiency Atrazine degrading bacteria and an application and screening method thereof. The bacteria are ZXY-2, belong to Arthrobacter, are preserved in China General Microbiological Culture Collection Center and have the preservation number of CGMCC No.10937 and the preservation date of July 1st, 2015. The bacteria can grow with Atrazine as the unique carbon and nitrogen source, and Atrazine with the initial concentration of 100 mg/L can be completely degraded within 14 hours. The strain Arthrobacter sp. ZXY-2 has the optimal growth temperature of 30 DEG C to 35 DEG C, the optimal growth pH of 8.0 to 9.0 and the optimal growth table rotating speed of 100 r/min to 200 r/min. The bacteria can be used for rapidly repairing the pollution of Atrazine pesticide.

Description

One plant height effect Atrazine degradation bacterium and application and screening method
Technical field
The present invention relates to technical field of environmental microorganism, in particular to a kind of bacterium for G-30027 of degrading, screening method and application thereof.
Background technology
China is as large agricultural country, and the amount of application of agricultural chemicals increases year by year.In recent years, due to the irrational use agricultural chemicals of people, in addition the bio-refractory characteristic of agricultural chemicals, agricultural chemicals often flows in river and soil along with precipitation, severe contamination surface and ground water.G-30027, as the important weedicide of a kind of northern area, causes serious harm to nature animals and plants because extensively using.G-30027 how in efficient degradation sewage is the hot issue of current water pollutions process field.In order to obtain stability and high efficiency degradation effect, adopting microorganism remediation technology G-30027 to be decomposed into nontoxic or low toxicity material and there is development prospect widely.Wherein, the most key part is the function stem that screening has high-level efficiency degraded G-30027.
Summary of the invention
The object of the invention is to provide a kind of bacterium for G-30027 of degrading and screening method thereof, for the quick in situ by atrazine-contaminated area repairs based theoretical.
For realizing the object of the invention, provide following technical scheme: a plant height effect Atrazine degradation bacterium, it is characterized in that described bacterial strain is Arthrobactersp.ZXY-2, be preserved on June 1st, 2015 that " China Committee for Culture Collection of Microorganisms's common micro-organisms " center ", deposit number is CGMCCNo.10937.
Address: No. 3, Yard 1, BeiChen xi Road, Chaoyang District, Beijing City.
One plant height effect Atrazine degradation bacterium, is characterized in that the quick reparation of the water body that can be applicable to G-30027 pesticidal contamination.
For realizing the object of the invention, a kind of efficient Atrazine degradation bacterial screening method being provided, it is characterized in that comprising the following steps:
(1) get 10g insecticide factory polluted soil, add 100ml and contain in the triangular pyramidal bottle of the minimal medium of 100mg/L G-30027, on 30 DEG C of constant-temperature tables, 150r/min domestication is cultivated; Got domestication nutrient solution every 7 days and be seeded to fresh containing in the G-30027 minimal medium of 100mg/L, inoculum size is 10%(v/v);
(2), after continuous domestication cultivates 30 days, enrichment culture thing is diluted 10 respectively -1, 10 -2, 10 -3, 10 -4, 10 -5, 10 -6, 10 -7, 10 -8with 10 -9doubly, respectively get 0.1mL and coat respectively on 100mg/L G-30027 solid plate, be placed in 30 DEG C of constant incubators and cultivate 2-3 days;
(3) picking list bacterium colony carries out bacteria purification cultivation, repeats more than 3 times until flat board occurs single bacterium colony that form size is identical;
(4) after the single bacterium colony line in picking step (3) cultivates 2-3 days to inclined-plane solid medium, 4 DEG C of preservations.
As preferably, minimal medium consists of: KH 2pO 40.9g/L, Na 2hPO 4.12H 2o6.5g/L, sucrose 3g/L, MgSO 4.7H 2o0.2g/L, FeSO 4.7H 2o0.01g/L, and 1ml/L liquid microelement, wherein liquid microelement is: CoCl 2.6H 2o0.1g, MnCl 2.4H 2o0.425g, ZnCl 20.05g, NiCl 2.6H 2o0.01g, CuSO 4.5H 2o0.015g, Na 2moO 4.2H 2o0.01g, Na 2seO 4.2H 2o0.01g is dissolved in 1000ml distilled water.
As preferably, slant medium is: KH 2pO 40.9g/L, Na 2hPO 4.12H 2o6.5g/L, sucrose 3g/L, peptone 1g/L, yeast powder 0.5g/L, NaCl1g/L, MgSO 4.7H 2o0.2g/L, FeSO 4.7H 2o0.01g/L, 1ml/L liquid microelement and 15-20g/L agar.
As preferably, inorganic salt solid medium separately adds agar 15-20g/L in minimal medium.
As preferably, the initial pH of inorganic salt, slant medium is 8.0.
As preferably, inorganic salt, slant medium high pressure steam sterilization condition are 121 DEG C, 15min.
Bacterial strain of the present invention is Gram positive aerobic tyrothricin, and long 1.58 μm, wide 1.13 μm, atrichia, without gemma, bacterial strain grows on inorganic salt film solid media, forms opaque faint yellow bacterium colony and the smooth of the edge.This bacterium can grow using G-30027 as sole carbon nitrogenous source, and starting point concentration can be made to be that the G-30027 of 100mg/L is degradable in 14h.This bacterial strain optimum growth temperature is 30-35 DEG C, and optimal pH is 8.0-9.0, and the most suitable growth shaking speed is 100-200r/min.
Bacterial strain of the present invention is identified through 16SrDNA, and its result is carried out tetraploid rice by Blast program, and the homology of this bacterial strain and ArthrobacterTBD185 reaches 99%.Its sequence is as follows:
ACGCTGGCGGCGTGCTTAACACATGCAAGTCGAACGATGATCCGGTGCTTGCGCCGGGGATTAGTGGCGAACGGGTGAGTAACACGTGAGTAACCTGCCCTTGACTCTGGGATAAGCCTGGGAAACTGGGTCTAATACCGGATATGACTCCTCATCGCATGGTGGGGGGTGGAAAGCTTTTTGTGGTTTTGGATGGACTCGCGGCCTATCAGCTTGTTGGTGGGGTAATGGCCTACCAAGGCGACGACGGGTAGCCGGCCTGAGAGGGTGACCGGCCACACTGGGACTGAGACACGGCCCAGACTCCTACGGGAGGCAGCAGTGGGGAATATTGCACAATGGGCGAAAGCCTGATGCAGCGACGCCGCGTGAGGGATGACGGCCTTCGGGTTGTAAACCTCTTTCAGTAGGGAAGAAGCCCTCTTTGGGGGTGACGGTACTTGCAGAAGAAGCGCCGGCTAACTACGTGCCAGCAGCCGCGGTAATACGTAGGGCGCAAGCGTTATCCGGAATTATTGGGCGTAAAGAGCTCGTAGGCGGTTTGTCGCGTCTGCTGTGAAAGACCGGGGCTCAACTCCGGTTCTGCAGTGGGTACGGGCAGACTAGAGTGCAGTAGGGGAGACTGGAATTCCTGGTGTAGCGGTGAAATGCGCAGATATCAGGAGGAACACCGATGGCGAAGGCAGGTCTCTGGGCTGTAACTGACGCTGAGGAGCGAAAGCATGGGGAGCGAACAGGATTAGATACCCTGGTAGTCCATGCCGTAAACGTTGGGCACTAGGTGTGGGGGACATTCCACGTTTTCCGCGCCGTAGCTAACGCATTAAGTGCCCCGCCTGGGGAGTACGGCCGCAAGGCTAAAACTCAAAGGAATTGACGGGGGCCCGCACAAGCGGCGGAGCATGCGGATTAATTCGATGCAACGCGAAGAACCTTACCAAGGCTTGACATGGACCGGAAAGACCTGGAAACAGGTGCCCCGCTTGCGGCCGGTTTACAGGTGGTGCATGGTTGTCGTCAGCTCGTGTCGTGAGATGTTGGGTTAAGTCCCGCAACGAGCGCAACCCTCGTTCTATGTTGCCAGCGGTTCGGCCGGGGACTCATAGGAGACTGCCGGGGTCAACTCGGAGGAAGGTGGGGACGACGTCAAATCATCATGCCCCTTATGTCTTGGGCTTCACGCATGCTACAATGGCCGGTACAAAGGGTTGCGATACTGTGAGGTGGAGCTAATCCCAAAAAGCCGGTCTCAGTTCGGATTGGGGTCTGCAACTCGACCCCATGAAGTCGGAGTCGCTAGTAATCGCAGATCAGCAACGCTGCGGTGAATACGTTCCCGGGCCTTGTACACACCGCCCGTCAAGTCACGAAAGTTGGTAACACCCGAAGCCGGTGGCCTAACCCGTGTGGGGGGAGCCGTCGAAGGTGG
Beneficial effect of the present invention: Atrazine-degrading Strains Arthrobactersp.ZXY-2 provided by the invention can using G-30027 as sole carbon nitrogenous source, for Study on Atrazine Biodegradation point-source pollution is increased economic efficiency.Starting point concentration can be that the G-30027 of 100mg/L is degradable in 14h by Atrazine-degrading Strains Arthrobactersp.ZXY-2 provided by the invention.
Accompanying drawing explanation
Fig. 1 is strains A rthrobactersp.ZXY-2 atomic force microscope figure.
Fig. 2 is strains A rthrobactersp.ZXY-2 growth curve chart.
Fig. 3 is strains A rthrobactersp.ZXY-2 degraded G-30027 graphic representation.
Fig. 4 degraded figure that is strains A rthrobactersp.ZXY-2 under with or without sucrose condition.
Fig. 5 is strains A rthrobactersp.ZXY-2 Atrazine degradation figure at different temperatures.
Fig. 6 is strains A rthrobactersp.ZXY-2 Atrazine degradation figure under different pH condition.
Fig. 7 is strains A rthrobactersp.ZXY-2 Atrazine degradation figure under different shaking speed condition.
Fig. 8 is strains A rthrobactersp.ZXY-2 degraded figure under different initial atrazine concentrations condition.
Embodiment
Embodiment 1: bacterial strain of the present invention screens from Jilin insecticide factory of the Chemical Co., Ltd. soil of long-term application G-30027, and its screening mode realizes according to the following steps.
(1) get 10g Jilin insecticide factory of Chemical Co., Ltd. polluted soil, add 100ml and contain in the triangular pyramidal bottle of minimal medium (pH=8) of 100mg/L G-30027, on 30 DEG C of constant-temperature tables, 150r/min domestication is cultivated.Got domestication nutrient solution every 7 days and be seeded to fresh containing in the G-30027 minimal medium of 100mg/L, inoculum size is 10%(v/v);
(2), after continuous domestication cultivates 30 days, enrichment culture thing is diluted 10 respectively -1, 10 -2, 10 -3, 10 -4, 10 -5, 10 -6, 10 -7, 10 -8with 10 -9doubly, respectively get 0.1mL and coat respectively on 100mg/L G-30027 solid plate, be placed in 30 DEG C of constant incubators and cultivate 2-3 days;
(3) picking list bacterium colony carries out bacteria purification cultivation, repeats more than 3 times until flat board occurs single bacterium colony that form size is identical;
(4) after the single bacterium colony line in picking step (3) cultivates 2-3 days to inclined-plane solid medium, 4 DEG C of preservations.
Wherein, minimal medium consists of: KH 2pO 40.9g/L, Na 2hPO 4.12H 2o6.5g/L, sucrose 3g/L, MgSO 4.7H 2o0.2g/L, FeSO 4.7H 2o0.01g/L, and 1ml/L liquid microelement.
Liquid microelement is: CoCl 2.6H 2o0.1g, MnCl 2.4H 2o0.425g, ZnCl 20.05g, NiCl 2.6H 2o0.01g, CuSO 4.5H 2o0.015g, Na 2moO 4.2H 2o0.01g, Na 2seO 4.2H 2o0.01g is dissolved in 1000ml distilled water.
Slant medium is: KH 2pO 40.9g/L, Na 2hPO 4.12H 2o6.5g/L, sucrose 3g/L, peptone 1g/L, yeast powder 0.5g/L, NaCl1g/L, MgSO 4.7H 2o0.2g/L, FeSO 4.7H 2o0.01g/L, 1ml/L liquid microelement and 15-20g/L agar.
Inorganic salt solid medium separately adds agar 15-20g/L in minimal medium
Above substratum is all at 121 DEG C, and autoclaving used after 15 minutes.
This bacterial strain is Gram positive aerobic bacterium, atrichia, and without gemma, bacterial strain grows on inorganic salt film solid media, forms opaque faint yellow bacterium colony and the smooth of the edge.
This bacterial strain transmission electron microscope picture as shown in Figure 1, is tyrothricin, long 1.58 μm, wide 1.13 μm.
Embodiment 2:
Use DNA extraction kit (Sangon) to extract DNA of bacteria according to extraction step, then carry out PCR object fragment amplification, PCR reaction conditions is: 94 DEG C of denaturation 5min, again through 94 DEG C of sex change 1min of 30 circulations, 55 DEG C of annealing 1min, 72 DEG C extend 1.5min, then 72 DEG C extend 5min.Pcr amplification product delivers to the order-checking of the precious biotech firm in Dalian.This bacterial strain is identified through 16SrDNA, and its result is carried out tetraploid rice by Blast program, and the homology of this bacterial strain and ArthrobacterTBD185 reaches 99%.Its sequence is as follows:
ACGCTGGCGGCGTGCTTAACACATGCAAGTCGAACGATGATCCGGTGCTTGCGCCGGGGATTAGTGGCGAACGGGTGAGTAACACGTGAGTAACCTGCCCTTGACTCTGGGATAAGCCTGGGAAACTGGGTCTAATACCGGATATGACTCCTCATCGCATGGTGGGGGGTGGAAAGCTTTTTGTGGTTTTGGATGGACTCGCGGCCTATCAGCTTGTTGGTGGGGTAATGGCCTACCAAGGCGACGACGGGTAGCCGGCCTGAGAGGGTGACCGGCCACACTGGGACTGAGACACGGCCCAGACTCCTACGGGAGGCAGCAGTGGGGAATATTGCACAATGGGCGAAAGCCTGATGCAGCGACGCCGCGTGAGGGATGACGGCCTTCGGGTTGTAAACCTCTTTCAGTAGGGAAGAAGCCCTCTTTGGGGGTGACGGTACTTGCAGAAGAAGCGCCGGCTAACTACGTGCCAGCAGCCGCGGTAATACGTAGGGCGCAAGCGTTATCCGGAATTATTGGGCGTAAAGAGCTCGTAGGCGGTTTGTCGCGTCTGCTGTGAAAGACCGGGGCTCAACTCCGGTTCTGCAGTGGGTACGGGCAGACTAGAGTGCAGTAGGGGAGACTGGAATTCCTGGTGTAGCGGTGAAATGCGCAGATATCAGGAGGAACACCGATGGCGAAGGCAGGTCTCTGGGCTGTAACTGACGCTGAGGAGCGAAAGCATGGGGAGCGAACAGGATTAGATACCCTGGTAGTCCATGCCGTAAACGTTGGGCACTAGGTGTGGGGGACATTCCACGTTTTCCGCGCCGTAGCTAACGCATTAAGTGCCCCGCCTGGGGAGTACGGCCGCAAGGCTAAAACTCAAAGGAATTGACGGGGGCCCGCACAAGCGGCGGAGCATGCGGATTAATTCGATGCAACGCGAAGAACCTTACCAAGGCTTGACATGGACCGGAAAGACCTGGAAACAGGTGCCCCGCTTGCGGCCGGTTTACAGGTGGTGCATGGTTGTCGTCAGCTCGTGTCGTGAGATGTTGGGTTAAGTCCCGCAACGAGCGCAACCCTCGTTCTATGTTGCCAGCGGTTCGGCCGGGGACTCATAGGAGACTGCCGGGGTCAACTCGGAGGAAGGTGGGGACGACGTCAAATCATCATGCCCCTTATGTCTTGGGCTTCACGCATGCTACAATGGCCGGTACAAAGGGTTGCGATACTGTGAGGTGGAGCTAATCCCAAAAAGCCGGTCTCAGTTCGGATTGGGGTCTGCAACTCGACCCCATGAAGTCGGAGTCGCTAGTAATCGCAGATCAGCAACGCTGCGGTGAATACGTTCCCGGGCCTTGTACACACCGCCCGTCAAGTCACGAAAGTTGGTAACACCCGAAGCCGGTGGCCTAACCCGTGTGGGGGGAGCCGTCGAAGGTGG
Embodiment 3:
Picking 4 DEG C of inclined-planes are preserved bacterium Arthrobactersp.ZXY-2 mono-ring and be cultured to 10 to 100mg/L G-30027 liquid nutrient mediums 8during MPN/ml, according to 5%(v/v) the bacterium amount that connects bacterial suspension is seeded in fresh inorganic salt liquid substratum, inoculation be placed on 30 DEG C, cultivate in the shaking table of 150r/min.Wherein, liquid nutrient medium initial pH value is 8.0.Take time as X-coordinate, absorbancy OD 600value is ordinate zou, draws growth curve.Period is every 1h sampling and measuring OD 600value.Growth curve of bacteria figure as shown in Figure 2, the adjustment period that this bacterium being in 0h-4h; Logarithmic phase is entered from 4h; During 18h, OD 600value is 0.8; When bacterium continued growth is to 24h, OD 600value is 1.0.
Embodiment 4:
Picking 4 DEG C of inclined-planes are preserved bacterium Arthrobactersp.ZXY-2 mono-ring and be cultured to 10 to 100mg/L G-30027 liquid nutrient mediums 8during MPN/ml, according to 5%(v/v) the bacterium amount that connects bacterial suspension is seeded in fresh inorganic salt liquid substratum, inoculation be placed on 30 DEG C, cultivate in the shaking table of 150r/min.Wherein, liquid nutrient medium initial pH value is 8.0.Take time as X-coordinate, atrazine concentration is ordinate zou, draws degradation by bacteria curve.Period samples every 1h, and after centrifugal for bacterial suspension 10000r/min 10min, filter with 0.22 μm of water system filter, utilize the concentration of high-performance liquid chromatogram determination G-30027, chromatographic condition is as follows:
Moving phase is acetonitrile: water=6:4(V/V), determined wavelength is 220nm, column temperature 30 DEG C, flow velocity 1ml/min, the long C18 post of sample size 20 μ L, 25cm.
As shown in Figure 3, bacterial strain can by degradable for the G-30027 of 100mg/L in 14h for degradation by bacteria graphic representation.
Embodiment 5:
Picking 4 DEG C of inclined-planes preservation bacterium Arthrobactersp.ZXY-2 mono-rings are cultivated, when bacterial growth to 10 to 100mg/L containing in the G-30027 liquid nutrient medium of sucrose and without in the G-30027 liquid nutrient medium of sucrose respectively 8during MPN/ml, respectively according to 5%(v/v) the bacterium amount that connects bacterial suspension is seeded to fresh in the inorganic salt liquid substratum of sucrose and without in the inorganic salt liquid substratum of sucrose, inoculation is placed on 30 DEG C, cultivates 24h in the shaking table of 150r/min.Wherein, liquid nutrient medium initial pH value is 8.0.Being X-coordinate with time, take atrazine degradation rate as ordinate zou, and investigate the degradation rate of G-30027 after 24h, its detection method is with embodiment 4.
This bacterium degrades figure as shown in Figure 4 under with or without sucrose condition, and within the 24h time, bacterial strain all by degradable for the G-30027 of 100mg/L, can illustrate that this bacterium can grow using G-30027 as sole carbon nitrogenous source under with or without sucrose condition.
Embodiment 6:
Picking 4 DEG C of inclined-planes are preserved bacterium Arthrobactersp.ZXY-2 mono-ring and be cultured to 10 to 100mg/L G-30027 liquid nutrient mediums 8during MPN/ml, according to 5%(v/v) the bacterium amount that connects bacterial suspension is seeded in fresh inorganic salt liquid substratum, inoculation be placed in 5 DEG C, 10 DEG C, 15 DEG C, 20 DEG C, 25 DEG C, 30 DEG C, 35 DEG C, 40 DEG C shaking tables, 150r/min cultivates 24h.Wherein, liquid nutrient medium initial pH value is 8.0.Take time as X-coordinate, atrazine degradation rate is ordinate zou, and the degradation rate of G-30027 under investigation condition of different temperatures, its detection method is with embodiment four.
Under different temperature condition to the degradation rate of G-30027 as shown in Figure 5, bacterial strain degradation capability within the scope of 30 DEG C-35 DEG C is the strongest, can reach 100% for this bacterium; Lower than 30 DEG C or higher than 35 DEG C its degraded G-30027s abilities all slightly decline.
Embodiment 7:
Picking 4 DEG C of inclined-planes are preserved bacterium Arthrobactersp.ZXY-2 mono-ring and be cultured to 10 to 100mg/L G-30027 liquid nutrient mediums 8during MPN/ml, according to 5%(v/v) the bacterium amount that connects bacterial suspension is seeded in fresh inorganic salt liquid substratum, wherein, liquid nutrient medium initial pH value is respectively 4.0,5.0,6.0,7.0,8.0,9.0,10.0.Inoculation is placed in 30 DEG C of shaking tables, and 150r/min cultivates 24h.Take time as X-coordinate, atrazine degradation rate is ordinate zou, and the degradation rate of G-30027 under investigation different pH condition, its detection method is with specific embodiment 4.
Under different pH value condition to the degradation rate of G-30027 as shown in Figure 6, bacterial strain degradation capability within the scope of pH8.0-pH9.0 is the strongest, can reach 100% for this bacterium; During lower than pH8.0 or higher than pH9.0, the ability of its degraded G-30027 all slightly declines.
Embodiment 8:
Picking 4 DEG C of inclined-planes are preserved bacterium Arthrobactersp.ZXY-2 mono-ring and be cultured to 10 to 100mg/L G-30027 liquid nutrient mediums 8during MPN/ml, according to 5%(v/v) the bacterium amount that connects bacterial suspension is seeded in fresh inorganic salt liquid substratum, wherein, liquid nutrient medium initial pH value is 8.0.Inoculation is placed in 30 DEG C of shaking tables, and 0r/min, 50r/min, 100r/min, 150r/min, 200r/min cultivate 24h respectively.Take time as X-coordinate, atrazine degradation rate is ordinate zou, and the degradation rate of G-30027 under investigation different pH condition, its detection method is with specific embodiment 4.
Under different shaking speed condition to the degradation rate of G-30027 as shown in Figure 7, bacterial strain degradation capability when 100r/min-200r/min is the strongest, can reach 100% for this bacterium; During lower than 100r/min, the ability of its degraded G-30027 all slightly declines.
Embodiment 9:
Picking 4 DEG C of inclined-planes are preserved bacterium Arthrobactersp.ZXY-2 mono-ring and be cultured to 10 to 100mg/L G-30027 liquid nutrient mediums 8during MPN/ml, according to 5%(v/v) the bacterium amount that connects bacterial suspension is seeded to respectively in the fresh inorganic salt liquid substratum of 10mg/L, 20mg/L, 50mg/L, 100mg/L, 200mg/L and 500mg/L, inoculation is placed in 30 DEG C of shaking tables, and 150r/min cultivates 24h.Wherein, liquid nutrient medium initial pH value is 8.0.Take time as X-coordinate, atrazine degradation rate is ordinate zou, and investigate the degradation rate under different initial atrazine concentrations condition, its detection method is with embodiment four.
This bacterium under different initial atrazine concentrations conditions to the degradation rate of G-30027 as shown in Figure 8, all can all degrade by bacterial strain within the scope of 10mg/L-200mg/L by G-30027.

Claims (8)

1. a plant height effect Atrazine degradation bacterium, is characterized in that described bacterial strain is Arthrobactersp.ZXY-2, is preserved in that " China Committee for Culture Collection of Microorganisms's common micro-organisms " center ", deposit number is CGMCCNo.10937 on June 1st, 2015.
2. a plant height effect Atrazine degradation bacterium according to claim 1, is characterized in that the quick reparation of the water body that can be applicable to G-30027 pesticidal contamination.
3. the efficient Atrazine degradation bacterial screening method as described in one of claim 1 ~ 2, is characterized in that comprising the following steps:
Get 10g insecticide factory polluted soil, add 100ml and contain in the triangular pyramidal bottle of the minimal medium of 100mg/L G-30027, on 30 DEG C of constant-temperature tables, 150r/min domestication is cultivated; Got domestication nutrient solution every 7 days and be seeded to fresh containing in the G-30027 minimal medium of 100mg/L, inoculum size is 10%(v/v);
After continuous domestication cultivates 30 days, enrichment culture thing is diluted 10 respectively -1, 10 -2, 10 -3, 10 -4, 10 -5, 10 -6, 10 -7, 10 -8with 10 -9doubly, respectively get 0.1mL and coat respectively on 100mg/L G-30027 solid plate, be placed in 30 DEG C of constant incubators and cultivate 2-3 days;
Picking list bacterium colony carries out bacteria purification cultivation, repeats more than 3 times until flat board occurs single bacterium colony that form size is identical;
After single bacterium colony line in picking step (3) cultivates 2-3 days to inclined-plane solid medium, 4 DEG C of preservations.
4. the efficient Atrazine degradation bacterial screening method of one according to claim 3, is characterized in that minimal medium consists of: KH 2pO 40.9g/L, Na 2hPO 4.12H 2o6.5g/L, sucrose 3g/L, MgSO 4.7H 2o0.2g/L, FeSO 4.7H 2o0.01g/L, and 1ml/L liquid microelement, wherein liquid microelement is: CoCl 2.6H 2o0.1g, MnCl 2.4H 2o0.425g, ZnCl 20.05g, NiCl 2.6H 2o0.01g, CuSO 4.5H 2o0.015g, Na 2moO 4.2H 2o0.01g, Na 2seO 4.2H 2o0.01g is dissolved in 1000ml distilled water.
5. the efficient Atrazine degradation bacterial screening method of one according to claim 3, is characterized in that slant medium is: KH 2pO 40.9g/L, Na 2hPO 4.12H 2o6.5g/L, sucrose 3g/L, peptone 1g/L, yeast powder 0.5g/L, NaCl1g/L, MgSO 4.7H 2o0.2g/L, FeSO 4.7H 2o0.01g/L, 1ml/L liquid microelement and 15-20g/L agar.
6. the efficient Atrazine degradation bacterial screening method of one according to claim 3, is characterized in that inorganic salt solid medium separately adds agar 15-20g/L in minimal medium.
7. the efficient Atrazine degradation bacterial screening method of one according to claim 3, is characterized in that the initial pH of inorganic salt, slant medium is 8.0.
8. the efficient Atrazine degradation bacterial screening method of one according to claim 3, is characterized in that inorganic salt, slant medium high pressure steam sterilization condition is 121 DEG C, 15min.
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CN105950504A (en) * 2016-05-19 2016-09-21 哈尔滨工业大学 Culture medium for atrazine degradation bacteria ZXY-2
CN109182173A (en) * 2018-09-07 2019-01-11 山东省科学院生态研究所 The application of arthrobacterium DnL1-1 and its microbial inoculum and preparation method
CN109234187A (en) * 2018-08-15 2019-01-18 李晓明 One plant of atrazine degradation bacteria and its application
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CN111518715A (en) * 2020-04-02 2020-08-11 哈尔滨工业大学(深圳)(哈尔滨工业大学深圳科技创新研究院) Sulfonamide antibiotic synergistic degradation bacteria and application thereof
CN112175882A (en) * 2020-10-21 2021-01-05 康生元(肇庆)生物科技有限公司 Bacterial strain KY331, microbial inoculum, product containing microbial inoculum and application of microbial inoculum

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Publication number Priority date Publication date Assignee Title
CN105950504A (en) * 2016-05-19 2016-09-21 哈尔滨工业大学 Culture medium for atrazine degradation bacteria ZXY-2
CN105950504B (en) * 2016-05-19 2019-11-05 哈尔滨工业大学 The culture medium of Atrazine degradation bacterium ZXY-2
CN109234187A (en) * 2018-08-15 2019-01-18 李晓明 One plant of atrazine degradation bacteria and its application
CN109234187B (en) * 2018-08-15 2021-06-15 李晓明 Atrazine degrading bacterium and application thereof
CN109182173A (en) * 2018-09-07 2019-01-11 山东省科学院生态研究所 The application of arthrobacterium DnL1-1 and its microbial inoculum and preparation method
CN109182173B (en) * 2018-09-07 2021-05-25 山东省科学院生态研究所 Application of arthrobacterium DnL1-1, microbial inoculum and preparation method thereof
CN111073835A (en) * 2019-12-28 2020-04-28 河南省农业科学院植物保护研究所 Pseudomonas mendocina capable of effectively degrading atrazine and application thereof
CN111518715A (en) * 2020-04-02 2020-08-11 哈尔滨工业大学(深圳)(哈尔滨工业大学深圳科技创新研究院) Sulfonamide antibiotic synergistic degradation bacteria and application thereof
CN111518715B (en) * 2020-04-02 2021-09-28 哈尔滨工业大学(深圳)(哈尔滨工业大学深圳科技创新研究院) Sulfonamide antibiotic synergistic degradation bacteria and application thereof
CN112175882A (en) * 2020-10-21 2021-01-05 康生元(肇庆)生物科技有限公司 Bacterial strain KY331, microbial inoculum, product containing microbial inoculum and application of microbial inoculum
CN112175882B (en) * 2020-10-21 2022-07-19 康生元(肇庆)生物科技有限公司 Bacterial strain KY331, microbial inoculum, product containing microbial inoculum and application of microbial inoculum

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