CN109136097A - The penicillium oxalicum of degradation isopropyl methoxalamine and its application - Google Patents

The penicillium oxalicum of degradation isopropyl methoxalamine and its application Download PDF

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CN109136097A
CN109136097A CN201810762390.4A CN201810762390A CN109136097A CN 109136097 A CN109136097 A CN 109136097A CN 201810762390 A CN201810762390 A CN 201810762390A CN 109136097 A CN109136097 A CN 109136097A
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isopropyl methoxalamine
penicillium oxalicum
degradation
plant
met
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CN109136097B (en
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孙扬
李永涛
赵丽霞
郝月崎
徐会娟
李晓晶
周斌
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Agro Environmental Protection Institute Ministry of Agriculture
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    • C02F3/347Use of yeasts or fungi
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    • C02TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02FTREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
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Abstract

The present invention relates to one plant degrade isopropyl methoxalamine penicillium oxalicum and its application, strain name is MET-F-1, classification naming are as follows: penicillium oxalicum Penicillium oxalicum, depositary institution: China Committee for Culture Collection of Microorganisms's common micro-organisms center, deposit number are CGMCC No.15861.The rDNA-ITS sequence of bacterial strain MET-F-1 of the present invention is by 414 base compositions, Co metabolism is carried out to herbicide isopropyl methoxalamine as single bacterial strain and adequate nutrition substance to the Penicillium notatum Penicillium oxalicum that screening obtains, determine the variation of isopropyl methoxalamine residual concentration in continuous 16d incubation, the results showed that penicillium oxalicum MET-F-1 and adequate nutrition substance can degrade isopropyl methoxalamine well.

Description

The penicillium oxalicum of degradation isopropyl methoxalamine and its application
Technical field
The invention belongs to microorganism fields, are related to a kind of degrading microorganism of herbicide, are related to one plant of degradation isopropyl first grass The penicillium oxalicum of amine and its application.
Background technique
Chloroacetanilide herbicides are the main organic pollutant of current China's agricultural land soil and water environment, especially isopropyl Alachlor etc. is difficult to environmental problem caused by the kind of mineralising and human health problems are got worse.Keep clean soil and Aquatic ecosystem realizes Agricultural Clean, the lasting safe utilization of soil and water resource becomes urgently to be resolved great in China One of agricultural, environment and food-safety problem.The various Method means for solving such pollution emerge one after another, wherein for microorganism The exploration of degradation chloroacetanilide herbicides has become hot spot in recent years.Mainstream product as current chloroacetanilide herbicides Kind, isopropyl methoxalamine is to prevent and kill off the annual gramineous weed in corn and soybean, sunflower, peanut and vegetable field, broadleaf class miscellaneous Grass and chufa application are universal.Stability of the isopropyl methoxalamine in the matrix such as soil and water is better than chloroacetyl amine weeding Other kinds of agent, such as Acetochlor, propachlor.Due to relatively high water solubility (550mg L-1), soil particle pair Its absorption is weaker, the risk that the isopropyl methoxalamine in soil has higher leaching to flow to surface water to underground water and diameter.Moreover, having Data show that isopropyl methoxalamine has certain bioaccumulation effect, and especially in edible Fish, this just gives human health band Carry out certain harm.
Current penicillium oxalicum is mainly used for control of plant disease, it is known that diseases prevention mechanism have secretion enzymatic hydrolysis substance, induction Resistance etc..Penicillium oxalicum metabolism, which generates, is put into the growth that prevent plant disease, pests, and erosion application prospect mass-energy inhibits a variety of disease fungus, such as hyphal cluster germ and bacterium Core germ.Also have been reported that discovery penicillium oxalicum has the function of Soluble phosphorus, degrading pesticide and antibiotic and has to quality of agricultural product Improve the effect with growth-promoting.
Summary of the invention
The purpose of the present invention is to provide one plant degrade isopropyl methoxalamine penicillium oxalicum and its application, the present invention analyze survey Tried by the pure bacterium of the bacterial strain outside plus under the effect of the Co metabolism of nutriment to the degradation of herbicide isopropyl methoxalamine, this bacterium Strain has the prospect for the candidate strain repaired as the degradation of farmland herbicide isopropyl methoxalamine.
The present invention realizes that the technical solution of purpose is as follows:
The penicillium oxalicum of one plant of degradation isopropyl methoxalamine, strain name MET-F-1, classification naming are as follows: penicillium oxalicum Penicillium oxalicum, depositary institution: China Committee for Culture Collection of Microorganisms's common micro-organisms center, preservation Number be CGMCC No.15861, preservation date are as follows: on May 24th, 2018, preservation address are as follows: BeiChen West Road, Chaoyang District, BeiJing City 1 Number institute 3.
Moreover, the rDNA-ITS sequence of bacterial strain MET-F-1 is shown in sequence 1.
A kind of isopropyl methoxalamine degradation reagent, wraps the penicillium oxalicum of one plant of degradation isopropyl methoxalamine.
Moreover, containing additional carbon and/or nitrogen source.
Moreover, at least one containing glucose, yeast extract.
Application of the penicillium oxalicum of one plant of degradation isopropyl methoxalamine as degradation isopropyl methoxalamine.
Application of the penicillium oxalicum of one plant of degradation isopropyl methoxalamine as soil-repairing agent.
Application of the penicillium oxalicum of one plant of degradation isopropyl methoxalamine as water remediation agent.
The advantages and positive effects of the present invention are:
The present invention has carried out minimal medium mesoxalic acid mould joint nutriment glucose or/and yeast extract Co metabolism The degradation repairing research of the normal spraying herbicide isopropyl methoxalamine in farmland determines during culture 384h (16d) isopropyl in culture medium The variation of alachlor.The preliminary proof Penicillium notatum can be used as degradation bacteria strains come isopropyl methoxalamine of degrading, and can be used as farmland soil The candidate strain of earth herbicide isopropyl methoxalamine pollution amelioration.
The present invention is confirmed through overtesting: the culture of bacterial strain MET-F-1 is just are as follows: 0.2% glucose, 0.2% yeast extract and Co metabolism 50mg L in the presence of+0.1% yeast extract of 0.1% glucose-1Isopropyl methoxalamine, when cultivating 16d, MET-F-1+ + 0.1% yeast extract of 0.2% glucose (MFG), MET-F-1+0.2% yeast extract (MFY) and MET-F-1+0.1% glucose (MFGY) degradation rate of isopropyl methoxalamine is respectively up to 66.1%, 53.7% and 86.6% in processing, blank (CK) and control (MG And MY) processing in isopropyl methoxalamine degradation rate less than 32.1%.And only add isopropyl methoxalamine in the processing (MF) of fungi Degradation rate is only 30.3%.
Detailed description of the invention
Fig. 1 is colonial morphology and microscopic features of the fungal bacterial strain of the present invention on malt extract medium;
Fig. 2 be isopropyl methoxalamine blank basal salt media and add nutriment control in degradation curve;
Fig. 3 is the isopropyl methoxalamine degradation curve under fungal bacterial strain of the present invention and different nutriments Co metabolism;
Fig. 4 is degradation rate of the isopropyl methoxalamine in blank basal salt media and different nutriments control;
Fig. 5 is the isopropyl methoxalamine degradation rate under fungal bacterial strain of the present invention and different nutriments Co metabolism.
Specific embodiment
The method of the present invention is described below by specific embodiment.Unless stated otherwise, technology used in the present invention Means are method known in those skilled in the art.In addition, embodiment is interpreted as illustrative, rather than this hair is limited Bright range, the spirit and scope of the invention are limited only by the claims that follow.To those skilled in the art, it is not carrying on the back Under the premise of from spirit and scope of the present invention, in these embodiments material component and dosage carry out various changes or change It is dynamic to also belong to protection scope of the present invention.
For the present invention from Tianjin insecticide factory place soil, an isolated penicillium has carried out morphology respectively It is identified with molecular biology method.Morphological analysis includes cultural method, tabletting dyeing and small culture identification, molecular biology side Method selects ITS (the internal transcribed spacer region of the rRNA gene, i.e. genetic transcription Spacer region) and 'beta '-tubulin gene (TUB), it is sequenced by amplification, the results show that with penicillium oxalicum Penicillium Oxalicum rDNA ITS gene very high homology.Classification naming are as follows: Penicillium oxalicum, depositary institution: China is micro- Biological inoculum preservation administration committee common micro-organisms center, deposit number are CGMCC No.15861, preservation date are as follows: 2018 May 24, preservation address are as follows: Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3.
The application has studied isopropyl methoxalamine in penicillium oxalicum bacterial strain MET-F-1 using minimal medium under lab It is such fungi future in farmland pollution soil with the degradation situation under nutriment glucose and the effect of yeast extract Co metabolism And the application in water body provides solid scientific basis.
The screening of above-mentioned bacterial strains and physiological property test process are as follows:
1. materials and methods
1.1 samples, instrument, reagent and culture medium
Sample source: separation screening strain collecting soil sample is from Tianjin insecticide factory place.
Instrument consumptive material: the culture experiment of fungus degrading isopropyl methoxalamine is carried out using constant-temperature table (Jintan Ward, Changzhou). Sample takes supernatant to use ultra high efficiency after 0.22 μm of miillpore filter (Pall, the U.S.) filtering after being centrifuged (Beckman, Germany) Liquid chromatograph (H-Class, Waters, the U.S.) measures the content of isopropyl methoxalamine in culture medium extracting solution.
Reagent: RAidlab DN14 fungal gene group extracts kit, purchased from the prosperous limited public affairs of biotechnology share of Hangzhou sky Department;PCRMIX enzyme buffer liquid RAidlab PC0902 is purchased from Guangzhou Dongsheng Biotechnology Co., Ltd;Peptone, beef extract, ferment Female cream and agar powder are biological reagent, are purchased from the extensive and profound in meaning star biotechnology in Beijing Co., Ltd;Ammonium sulfate, phosphoric acid hydrogen two The reagents such as potassium, potassium dihydrogen phosphate, sodium chloride, epsom salt, sodium nitrate, potassium chloride, ferrous sulfate, sucrose and glucose are point Analyse it is pure, be purchased from Tianjin Feng Chuan Chemical Co., Ltd..
Enriched medium: peptone 5g/L, beef extract 3g/L, sodium chloride 5g/L, distilled water constant volume adjust pH to 7.0, use It is preceding high pressure sterilization 20 minutes at a temperature of 121 DEG C.
Basal salt media: ammonium sulfate 1.0g/L, dipotassium hydrogen phosphate 1.5g/L, potassium dihydrogen phosphate 0.5g/L, sodium chloride 1.0g/L, epsom salt 0.4g/L, distilled water constant volume, with hydrochloric acid tune pH to neutrality.It is gone out using preceding high pressure at a temperature of 121 DEG C Bacterium 20 minutes.
Pure medium (czapek agar medium): sodium nitrate 3.0g/L, dipotassium hydrogen phosphate 1.0g/L, epsom salt 0.5g/L, potassium chloride 0.5g/L, ferrous sulfate 0.01g/L, sucrose 30.0g/L, agar 20g/L, distilled water constant volume, after packing 121 DEG C of high pressure sterilization 20min, it is spare.
LB liquid medium: tryptone 10g/L, yeast extract 5g/L, NaCl10g/L, distilled water constant volume, tune pH is extremely 7.0, using preceding high pressure sterilization 20 minutes at a temperature of 121 DEG C.
Degradation experiment culture medium: same to basal salt media.
2 methods
2.1 bacterial strain screening
The pedotheque 10g derived from Tianjin insecticide factory place is fetched, under aseptic technique, is added to the grass of first containing isopropyl Amine concentration is to cultivate 5 days under 150r/min frequency of oscillation at 30 DEG C in the triangular flask of the 100mL enriched medium of 50mg/L; Then the triangle for the 100mL enriched medium that concentration containing isopropyl methoxalamine is 100mg/L is transferred to by 10% inoculum concentration In bottle, continue culture 5 days;It is rich that the 100mL that concentration containing isopropyl methoxalamine is 200mg/L is transferred to by 10% inoculum concentration again In the triangular flask for collecting culture medium, continue culture 5 days;Then the above-mentioned enriched medium bacterium solution of 1mL is taken to be added in 10mL centrifuge tube, Distilled water constant volume, dilutes 10 step by step-1、10-2、10-3、10-4、10-5, switching 0.1mL dilution is dense to isopropyl methoxalamine is contained respectively Degree is on the pure medium plate of 100mg/L, and spread plate is placed in 30 DEG C of constant incubators and cultivates 3 days, chooses not similar shape The single colonie of state feature is inoculated on the pure medium that concentration containing isopropyl methoxalamine is 100mg/L, using plate streak point It carry out not isolate and purify for 3 times.5 plants of the preferable single colonie bacterial strain of growing way on plate is chosen after purification, is stored in Czapek's agar In the slant tube of culture medium.
The screening of 2.2 isopropyl methoxalamine function degradation bacterias
5 plants of bacterial strains after purification are cultivated in LB liquid medium to logarithm period (logarithm period use measurement OD600 Value determines that it is logarithm period that OD600 value, which reaches 0.8), then with 10% inoculum concentration (in this experiment " 10% inoculum concentration " Refer to inoculation liquid and the volume ratio for being vaccinated culture medium) accessed the basic inorganic salt that concentration containing isopropyl methoxalamine is 50mg/L In culture solution, minimal medium has following processing: 0.2% glucose, 0.2% yeast extract, 0.1% Portugal are separately added into before sterilizing Grape+0.1% yeast extract of sugar, while setting and not connecing the culture solution of bacterium and compare, 3 repetitions of each sample treatment.After culture, inhale Supernatant 2mL is taken, 10mL acetonitrile, 2g sodium chloride is added, vortex 1min, 8000r/min are centrifuged 5min, Aspirate supernatant 1mL, mistake 0.22 μm of filter membrane measures the residual quantity of isopropyl methoxalamine with Ultra Performance Liquid Chromatography instrument.By screening, one plant is obtained to isopropyl first The excellent fungal bacterial strain of careless amine Co metabolism function, is named as MET-F-1.Specifically it is arranged and the results are shown in Table 1,2.
Under the meaning of 1 different disposal of table number in minimal medium isopropyl methoxalamine residual quantity and degradation rate
The residual quantity and degradation rate of isopropyl methoxalamine in 2 blank of table and different nutriments control minimal medium
The result shows that: to containing concentration be 50mg/L isopropyl methoxalamine basal salt media in be added 0.2% glucose, The nutriment of+0.1% yeast extract of 0.2% yeast extract and 0.1% glucose, after cultivating 384h, wherein isopropyl methoxalamine Concentration range is 33.7-38.6mg/L, and most degradation rate is 35.7mg/L (degradation rate 28.1%) phase in 32.0%, with CK Than not influenced significantly on the degradation of wherein isopropyl methoxalamine.
The residual quantity and degradation rate of the fungal bacterial strain of the present invention of table 3 and the isopropyl methoxalamine under different nutriments Co metabolism
The result shows that: and after being inoculated with MET-F-1, the degradation of isopropyl methoxalamine in the processing (MF) that nonnutritive substance is added Effect (culture 384h degradation rate is 30.3%) does not improve significantly compared with CK (culture 384h degradation rate is 28.1%).And The nutrition of 0.2% glucose (MFG), 0.2% yeast extract (MFY) and+0.1% yeast extract of 0.1% glucose (MFGY) is added After substance, the degradation of isopropyl methoxalamine accelerates obvious in basal salt media, after cultivating 384h, isopropyl methoxalamine residual quantity difference Degradation to 16.8mg/L, 23.0mg/L and 6.7mg/L, degradation effect promotes degree to be ordered as MFGY > MFY > MFG, cultivates 384h When be respectively 86.6%, 66.1% and 53.7% to the degradation rate of isopropyl methoxalamine
The residues detection method of 2.3 pesticide isopropyl methoxalamines
2.3.1 external standard method is used: with isometric acetonitrile: deionized water (1:1) that isopropyl methoxalamine standard solution is successively dilute It is interpreted into 0.05,0.1,0.2,0.5,1,2,5,10mg/L concentration, is then measured with Ultra Performance Liquid Chromatography instrument, each 1 μ L of sample introduction, 3 repetitions of each sample treatment, take the average value of peak area.Using concentration as abscissa, peak area is that ordinate does standard curve.
2.3.2 the liquid phase chromatogram condition of isopropyl methoxalamine: WatersACQUITYUPLC HSS T3 chromatographic column, 1.8 μm of * 2.1mm*100mm;Column oven temperature: 30 DEG C;Mobile phase: gradient elution is used, 0min: acetonitrile: ultrapure water (10:90), 1min: Acetonitrile: ultrapure water (10:90), 5min: acetonitrile: ultrapure water (70:30), 7min: acetonitrile: ultrapure water (10:90) keeps 2min; Wavelength: 230nm.
The residual quantity of isopropyl methoxalamine in culture solution is obtained by standard curve, as a result sees Fig. 2-3, and then pass through formula 1 The degradation rate for calculating isopropyl methoxalamine, is as a result shown in Fig. 4-5.
Formula 1: degradation rate (%)=(isopropyl methoxalamine in isopropyl methoxalamine concentration-processing culture solution in initial incubation liquid Residual concentration) isopropyl methoxalamine concentration × 100% in/initial incubation liquid
The identification of 2.4 isopropyl methoxalamine function degradation bacteria strains MET-F-1
Strain gene group DNA extract: isolated strains using malt extract medium in 30 DEG C incubator stationary culture 5 days, make It is carried out with RAidlab DN14 fungi extracts kit according to its specification method, operating process is as follows: scraping 0.2-0.5mg bacterium Silk, is put into 2.0ml centrifuge tube;Add 2 diameter 3-4mm beades and 100 μ l CTAB lysates, it is even in Retsch MM400 It is ground on pulp grinder twice, each 2min;400 μ l, 65 DEG C of water-bath preheating CTAB lysates are added, in 65 DEG C of water-baths after being mixed by inversion 0.5-1hr;Isometric chloroform is added: isoamyl alcohol (24:1) is gently mixed by inversion 50 times;12000rpm is centrifuged under room temperature 5min takes supernatant into new centrifuge tube;1.5 times of volume combination liquid are added into supernatant, are transferred to absorption after mixing In column, 12000rpm is centrifuged 30sec under room temperature, discards waste liquid;500 μ l mortifiers are added to remove liquid, 12000rpm room temperature item It is centrifuged 30sec under part, discards waste liquid;600 μ l rinsing liquids are added, 12000rpm is centrifuged 30sec under room temperature, discards waste liquid; Adsorption column is put back into sky collecting pipe, 12000rpm is centrifuged 2min under room temperature;Adsorption column be transferred to the 1.5ml of sterilizing from In heart pipe, opening is dried at room temperature for 5min;Add 50~100 μ l elution buffers, place 3-5min at room temperature, 12000rpm from Heart 30sec collect Genomic DNA solution, immediately use or -20 DEG C under the conditions of store it is spare.
Ribosomal gene (rDNA) sequence amplification sequencing: by PCR MIX buffer and primer (upstream primer ITS5:5 '- GGAAGTAAAAGTCGTAACAAG-3 ' and downstream primer ITS4:5 '-TCCTCC-GCTTATTGATATGC-3 ') solution taking-up, Melt 30 minutes on 4 DEG C or ice cube.
PCR amplification system (50 μ l): 19 μ l of ddH2O;25 μ l of PCRMIX enzyme buffer liquid;2 μ l of upstream primer;Downstream primer 2 μl;2 μ l of template (first 4 kinds, at mixed liquor is distributed into, after dispensing PCR reaction tube, add DNA profiling).
Pcr amplification reaction program: 95 DEG C of denaturation 3min, 94 DEG C of unwindings 1min, 54 DEG C of renaturation 40s, 72 DEG C of extension 1min expand Increase 35 circulation after 72 DEG C of extension 10min, product saves at 4 DEG C after the completion.
Beijing Qing Kexin industry Bioisystech Co., Ltd is transferred to carry out analysis sequencing the PCR product for meeting stripe size, Obtain the rDNA-ITS sequence composition of the bacterial strain:
Above-mentioned rDNA-ITS sequence is made of 414 bases (bp).
The observation of 2.5 colony morphology characteristics
MET-F-1 bacterial strain is grown comparatively fast on malt extract medium, 5 days 15~20mm of colony diameter, edge white, middle part Celadon, tiling, conidium structure largely generate, the flaking of spore face;The back side is light brown.The mitogenetic spore of microscopically observation Son stalk is elongated, and penicillus common 2~3 is verticillate, fork;Wall is smooth, colourless;Bottle stalk column, 7.5~12.4 × 2.3~3.5 μm; Conidium ellipse or wide ellipse, it is smooth or slightly coarse, concatenate, 3.2~8.3 × 2.5~5.0 μm.Bacterial strain MET-F-1's Colonial morphology and microscopic morphology are shown in Fig. 1.
2.5 bacterial strain MET-F-1 are accredited as a kind of new function bacterial strain
Sequencing result is carried out Blast with the GenBank in NCBI to compare, obtains MET-F-1 and Penicillium The homology of the ITS sequence of oxalicum is 99%.It is analyzed according to the morphological feature of bacterial strain MET-F-1 and rDNA-ITS sequence As a result, the bacterial strain is accredited as penicillium oxalicum (Penicillium oxalicum), it is named as penicillium oxalicum MET-F-1 (Penicillium oxalicum MET-F-1).Up to now, penicillium oxalicum still is found without researcher both at home and abroad (Penicillium oxalicum) has the function of the report of degradation isopropyl methoxalamine.Therefore, bacterial strain MET-F-1 belongs to one kind New function bacterial strain is preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center on May 24th, 2018, Deposit number is CGMCC No.15861.
Sequence table
<110>Inst. of Environment Protection & Scientific Research Monitor, Ministry of Agric
<120>penicillium oxalicum of degradation isopropyl methoxalamine and its application
<160> 3
<170> SIPOSequenceListing 1.0
<210> 1
<211> 414
<212> DNA
<213>the rDNa-ItS sequence (2 Ambystoma laterale x Ambystoma jeffersonianum) of bacterial strain
<400> 1
gagcacaaca cgacaattcc tcgtgtcgtg cttcgatcta atatgagaac caaactgact 60
caagattcag gcaaaccatt gctggtgagc acggccttga cggcgatggc cagtaagtca 120
acagtttggc cccccccccc ccccacgatg agatcttgac gacagagaat ggcggtctga 180
tattttcggt tcagctacaa cggttcctcc gacctccagc tggagcgcct gaacgtctac 240
ttcacccacg taagtcgtga aaaatgtctt cccaatcaca gaaatctccc aaacctgacc 300
attgaaacgt tcaattttcc aggctagcgg tgacaagtat gttccccgtg ccgttctcgt 360
cgacctggag cccggtacca tggacgctgt ccgtgccggt ccctttggca agct 414
<210> 2
<211> 21
<212> DNA
<213>upstream primer ITS5 (2 Ambystoma laterale x Ambystoma jeffersonianum)
<400> 2
ggaagtaaaa gtcgtaacaa g 21
<210> 3
<211> 20
<212> DNA
<213>downstream primer ITS4 (2 Ambystoma laterale x Ambystoma jeffersonianum)
<400> 3
tcctccgctt attgatatgc 20

Claims (8)

1. the penicillium oxalicum of one plant of degradation isopropyl methoxalamine, it is characterised in that: strain name MET-F-1, classification naming are as follows: grass Sour mould Penicillium oxalicum, depositary institution: in China Committee for Culture Collection of Microorganisms's common micro-organisms The heart, deposit number are CGMCC No.15861, preservation date are as follows: on May 24th, 2018, preservation address are as follows: Chaoyang District, Beijing City north The institute 3 of occasion West Road 1.
2. the penicillium oxalicum of degradation isopropyl methoxalamine according to claim 1, it is characterised in that: bacterial strain MET-F-1's RDNA-ITS sequence is shown in sequence 1.
The reagent 3. a kind of isopropyl methoxalamine is degraded, it is characterised in that: include one plant of degradation isopropyl methoxalamine described in claim 1 Penicillium oxalicum.
The reagent 4. isopropyl methoxalamine according to claim 3 is degraded, it is characterised in that: contain additional carbon and/or nitrogen source.
The reagent 5. isopropyl methoxalamine according to claim 3 is degraded, it is characterised in that: contain glucose, yeast extract at least one Kind.
6. application of the penicillium oxalicum of one plant of degradation isopropyl methoxalamine described in claim 1 as degradation isopropyl methoxalamine.
7. application of the penicillium oxalicum of one plant of degradation isopropyl methoxalamine described in claim 1 as soil-repairing agent.
8. application of the penicillium oxalicum of one plant of degradation isopropyl methoxalamine described in claim 1 as water remediation agent.
CN201810762390.4A 2018-07-12 2018-07-12 The penicillium oxalicum of degradation isopropyl methoxalamine and its application Expired - Fee Related CN109136097B (en)

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