CN104388355B - Bacterium for degrading herbicides acetochlor and Blazer Zs and application thereof - Google Patents

Bacterium for degrading herbicides acetochlor and Blazer Zs and application thereof Download PDF

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CN104388355B
CN104388355B CN201410705138.1A CN201410705138A CN104388355B CN 104388355 B CN104388355 B CN 104388355B CN 201410705138 A CN201410705138 A CN 201410705138A CN 104388355 B CN104388355 B CN 104388355B
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rhizobium
acetochlor
weeds
cgmcc
burn
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高淼
孙建光
孙颖飞
张磊
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Institute of Agricultural Resources and Regional Planning of CAAS
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Abstract

The invention discloses a bacterium for degrading herbicides acetochlor and Blazer Zs and application thereof. The bacterium for degrading herbicide acetochlor and Blazer Zs is Rhizobium sp. 198-R-47 which is preserved in China General Microbiological Culture Collection Center and has a preservation number of CGMCC No.9867. After being cultured for 7 days in an inorganic salt culture medium, the Rhizobium sp. 198-R-47 (CGMCC No.9867) has a degradation rate of 73.65% for 50mg/L acetochlor and has a degradation rate of 9.04% for 100mg/L Blazer Zs, which means that the strain is capable of degrading acetochlor and/or Blazer Zs and has a wide application prospect in restoration of soil polluted by herbicides acetochlor and/or Blazer Zs.

Description

Bacterium that one plant of degrading herbicide Acetochlor and weeds are burnt and application thereof
Technical field
The present invention relates to bacterium that in microorganism field, one plant of degrading herbicide Acetochlor and weeds are burnt and application thereof.
Background technology
Acetochlor, english common name acetochlor, chemical name be 2- ethyl -6 methyl-n- ethoxyl methyl-α - Chloro acetanilide.
Acetochlor is absorbability acetamide-group herbicides, is selective preemergence herbicide.Can be by the young shoot of weeds and young root Absorb, suppression weeds protein synthesis, and make weeds dead.It is mainly used in the crops such as soybean, peanut, corn, cotton and prevent and kill off one Year raw grassy weed and part broad leaved weed, have good preventive effect to Soybean Dodder, invalid to perennial weeds.
Weeds are burnt, also known as acifluorfen, English name acifluorfen, chemical name 5- (the chloro- α of 2-, α, α-trifluoro To toloxyl) 2- nitrobenzoic acid.
It is diphenyl ether herbicide that weeds are burnt, and belongs to contact herbicide, is protoporphyrin oxidase inhibitor.Generally adopt 25% aqua carries out the soil process of soybean before seedling table and after seedling through foliar spray mist, can effectively prevent and kill off Soybean Field broad leaved weed such as Siberian cocklebur, piemarker Fiber crops, black nightshade, amaranth, lamb's-quarters, knotweed, lead a cow, artemisiifolia, datura, purslane, acalypha copperleaf, beggar-ticks, weasel hemp nettle, dayflower etc., after seedling is early Phase process also can prevent and kill off the annual gramineous weed including green bristlegrass.After spray, soybean meeting damage to different degrees, performance For yellowing leaf, shrinkage, brown stain etc., but recover normal after 6～7d.
Acetochlor and weeds burn that the half-life is shorter, are considered as a kind of environmentally friendly agricultural chemical of low toxicity always, but in recent years Come frequent, excess or improper use, different degrees of pollution is caused to agricultural land soil and underground water etc., the danger to succession crop Evil is also increasingly severe.Research shows, applies Acetochlor and weeds in the environment in a large number and burns and cause certain residual toxicity, its The several years can be remained in underground water, surface water, enter in environment and be constantly enriched with vivo by food chain, fish are had Stronger toxicity, environmental pollution is very big.Especially Acetochlor can cause the endocrine disturbance of organism, causes sterile, not just Normal gender differences are even carcinogenic, are set to carcinogenic substance by EPA.
The degraded of nature herbicide residue relies primarily on Soil Microorganism to complete, but natural degradation is very slow. Therefore, targetedly screen high-effective microorganism bacterial strain, develop microorganism renovation agent, Herbicidal agent is accelerated by artificial infection It is a very necessary job and practicable approach that the degraded of residual eliminates farmland herbicide herbicide carryover.
Content of the invention
The technical problem to be solved is how degrading herbicide Acetochlor and weeds are burnt.
For solve above-mentioned technical problem, the invention provides one plant can with degrading herbicide Acetochlor and weeds burn thin Bacterium.
The bacterium that the present invention provides is rhizobium (rhizobium sp.) 198-r-47, and this bacterial strain is in October, 2014 (abbreviation cgmcc, address is: Beijing to be preserved within 28th China Committee for Culture Collection of Microorganisms's common micro-organisms center Chaoyang District North Star West Road 1 institute 3), deposit number is cgmcc no.9867.
It is a further object to provide a kind of microbial inoculum, the active component of this microbial inoculum is described rhizobium (rhizobium sp.)198-r-47 cgmcc no.9867.
Above-mentioned microbial inoculum can be following 1) or 2) described microbial inoculum:
1) it is used for the microbial inoculum that degrading herbicide Acetochlor and/or weeds are burnt；
2) it is used for rehabilitating soil Determination of Herbicide Acetochlor and/or weeds burn the microbial inoculum of pollution.
Described microbial inoculum can also include carrier.Described carrier can be solid carrier or liquid-carrier.Described solid carrier can For mineral material, vegetable material or macromolecular compound；Described mineral material can be clay, talcum, kaolin, montmorillonite, white At least one in carbon, zeolite, silica and diatomite；Described vegetable material can be at least in corn flour, bean powder and starch Kind；Described macromolecular compound can be polyvinyl alcohol and/or polyglycols.Described liquid-carrier can be organic solvent, vegetable oil, ore deposit Thing oil or water；Described organic solvent can be decane and/or dodecane.In described microbial inoculum, described active component can be to be cultured Living cells, presented in the mixture of the zymotic fluid of living cells, the filtrate of cell culture or cell and filtrate.Described group The formulation of compound can be multiple formulations, such as liquor, emulsion, suspending agent, pulvis, granule, wettable powder or water dispersible granules.
As needed, also can add surfactant (as polysorbas20, Tween 80 etc.), adhesive in described microbial inoculum, stablize Agent (as antioxidant), ph conditioning agent etc..
Described rhizobium (rhizobium sp.) 198-r-47 cgmcc no.9867 or with rhizobium (rhizobium Sp.) 198-r-47 cgmcc no.9867 is microbial inoculum the answering in degrading herbicide Acetochlor and/or weeds burn of active component With falling within protection scope of the present invention.
Described rhizobium (rhizobium sp.) 198-r-47 cgmcc no.9867 or with rhizobium (rhizobium Sp.) 198-r-47 cgmcc no.9867 burns dirt for the microbial inoculum of active component in rehabilitating soil Determination of Herbicide Acetochlor and/or weeds Application in dye falls within protection scope of the present invention.
It is also another object of the present invention to provide a kind of culture rhizobium (rhizobium sp.) 198-r-47 cgmcc The method of no.9867.
The method of culture rhizobium (rhizobium sp.) 198-r-47 cgmcc no.9867 provided by the present invention, Train in the culture medium for cultivating rhizobium including by rhizobium (rhizobium sp.) 198-r-47 cgmcc no.9867 Foster step.
A further object of the present invention is to provide one kind with rhizobium (rhizobium sp.) 198-r-47 cgmcc No.9867 is the preparation method of the microbial inoculum of active component.
The preparation method of above-mentioned microbial inoculum provided by the present invention, comprises the steps: described rhizobium (rhizobium Sp.) 198-r-47 cgmcc no.9867, as active component, obtains described microbial inoculum.
The present invention is to be subject to the farmland of pollution of herbicide to gather soil sample from for a long time, and the herbicide that therefrom separation screening obtains Acetochlor and/or weeds burn degradation bacteria rhizobium (rhizobium sp.) 198-r-47 cgmcc no.9867.It is demonstrated experimentally that This bacterial strain reaches 73.65% to 50mg/l Acetochlor degradation rate in 7 days in minimal medium；The fall that 100mg/l weeds are burnt Solution rate reaches 9.04%.This shows that this bacterial strain energy degrading herbicide Acetochlor and/or weeds are burnt, in rehabilitating soil herbicide second grass Amine and weeds are burnt pollution aspect and have broad application prospects.
Preservation explanation
Strain name: rhizobium
Latin name: (rhizobium sp.)
Strain number: 198-r-47
Preservation mechanism: China Committee for Culture Collection of Microorganisms's common micro-organisms center
Preservation mechanism abbreviation: cgmcc
Address: Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3
Preservation date: on October 28th, 2014
Collection is registered on the books numbering: cgmcc no.9867
Specific embodiment
Experimental technique used in following embodiments if no special instructions, is conventional method.
Material used, reagent etc. in following embodiments, if no special instructions, all commercially obtain.
In following embodiments, culture medium used is as follows:
Nitrogen-free fluid nutrient medium: solute and its concentration are sucrose 10g/l, nacl 0.12g/l, k2hpo4·3h2o 0.5g/ L, caco31g/l, mgso4·7h2o 0.2g/l；Solvent is distilled water；ph7.2.
Nitrogen-free solid medium: add the culture that agar obtains to its content for 15-20g/l in nitrogen-free fluid nutrient medium Base.
Beef extract-peptone fluid nutrient medium: solute and its concentration are beef extract 5g/l, peptone 10g/l, nacl5g/l； Solvent is distilled water；ph7.2.
Beef extract-peptone solid medium: solute and its concentration are beef extract 5g/l, peptone 10g/l, nacl5g/l, Agar 15-20g/l；Solvent is distilled water；ph7.2.
Inorganic salt liquid culture medium: solute and its concentration are nh4Cl 1.0g/l, kh2po40.5g/l, k2hpo41.5g/ L, mgso40.2g/l, nacl 0.5g/l；Solvent is distilled water；ph7.0.
Inorganic salts solid medium: adding agar to its content in inorganic salt liquid culture medium is 2% culture medium obtaining.
Rhizobium (rhizobium sp.) ld1616cgmcc no.7775 (abbreviation rhizobium in following embodiments (rhizobium sp.) ld1616) disclosed in Chinese invention patent document cn 103343100b.
Embodiment 1, the separation of rhizobium (rhizobium sp.) 198-r-47 cgmcc no.9867 and identification
First, Acetochlor/weeds burn the separation of degradation bacteria 198-r-47
1st, 10g pedotheque is added (to pick up from the agriculture that Chinese Heihe In The Heilongjiang River is subject to pollution of herbicide in 100ml sterilized water Field), shaken cultivation 30min makes dirty solution.Draw the above-mentioned dirty solution of 1ml to add in the test tube filling in 9ml sterilized water fully (now dilution factor is designated as 10 for mixing-1), from this test tube, then draw 1ml be added in another test tube filling 9ml sterilized water Mix, make 10 by that analogy-2、10-3、10-4、10-5Different dilution bacteria suspensions.Each dilution factor is taken 0.1ml uniform It is coated on nitrogen-free solid medium flat board, 28 DEG C of constant temperature quiescent culture 23 days.After bacterium colony is formed, in nitrogen-free solid culture Purifying agaric is carried out on base flat board.
2nd, Determination of Herbicide Acetochlor, weeds are burnt, the preliminary screening of Pu Shite and chlorimuronethyl degradation capability adopts flat board saturating Bright circle method.It is 1000mg/l that Acetochlor standard items (fluka Products) addition inorganic salts solid medium is made its content, obtains To Acetochlor flat board；Weeds are burnt standard items (fluka Products) adds inorganic salts solid medium to make its content be 1000mg/l, obtains weeds and burns flat board；Pu Shite standard items (fluka Products) addition inorganic salts solid medium is made it Content is 1000mg/l, obtains general applying dead falt sheet；Chlorimuron ethyl (fluka Products) is added the training of inorganic salts solid It is 1000mg/l that foster base makes its content, obtains chlorimuronethyl flat board.By Acetochlor flat board, weeds burn flat board, general apply dead falt sheet and Chlorimuronethyl flat board carries out subregion, and aspiration step 1 purifies the rhizobium in the bacteria suspension 10 μ l of every plant of bacterium obtaining and this laboratory The bacteria suspension 10 μ l (the thalline content of various bacterial strain bacteria suspensions is identical) of (rhizobium sp.) ld1616 dibbling to four kinds respectively On flat board (every plant of bacterium is repeated 3 times on a flat board), 28 DEG C of cultures, observations.Screen one plant in Acetochlor flat board and weeds Burn on flat board and all can form the bacterial strain of larger transparent circle, it is named as Acetochlor/weeds and burns degradation bacteria 198-r-47 by this bacterial strain. Acetochlor/weeds are burnt degradation bacteria 198-r-47 and apply dead falt sheet and chlorimuronethyl flat board all can not form transparent circle general.Rhizobium (rhizobium sp.) ld1616 only forms larger transparent circle on chlorimuronethyl flat board, applies dead falt sheet, Acetochlor flat board general Burn with weeds and transparent circle all can not be formed on flat board.Illustrate Acetochlor/weeds burn degradation bacteria 198-r-47 can degrade Acetochlor and Weeds burn it is impossible to degrade chlorimuronethyl and Pu Shite；Rhizobium (rhizobium sp.) ld1616 can degrade chlorimuronethyl, no Can degrade Pu Shite, Acetochlor and weeds are burnt.
Table 1. Acetochlors/weeds burn degradation bacteria 198-r-47 and rhizobium (rhizobium sp.) ld1616 removes to four kinds Careless agent degradation capability primary dcreening operation result
Strain number Pu Shite Acetochlor Chlorimuronethyl Weeds are burnt
Acetochlor/weeds burn degradation bacteria 198-r-47 - + - +
Rhizobium (rhizobium sp.) ld1616 - - + -
Note: "+" representing the larger transparent circle of formation, "-" indicates that no transparent circle is formed.
2nd, Acetochlor/weeds burn the identification of degradation bacteria 198-r-47
Separate and purify the Acetochlor/weeds obtaining from the following aspects authentication step one and burn degradation bacteria 198-r-47:
1st, Morphological Identification
Exponential phase will be in and bacterium colony size will be stable, above-mentioned steps one will separate and purify the Acetochlor/weeds obtaining Burn degradation bacteria 198-r-47 and carry out single bacterium colony state observation, the main size including bacterium colony, color, transparency, wettability, bacterium colony Surface state (whether flat, projection, fold, depression etc.), colony edge state (whether neat, irregular, radial etc.).
Result shows, above-mentioned steps one separate and purify the Acetochlor/weeds obtaining burns degradation bacteria 198-r-47 bacterium colony circle Shape is raised, milky, moistening unclarity, regular edges, diameter 0.8-1.5mm.
2nd, analysis of physio biochemical characteristics
Reference " common bacteria system identification handbook " (east show pearl, Cai Miaoying. common bacteria system identification handbook. Beijing: section Publishing house, 2011.) and " Microbiology Experiment " (Shen Ping, Fan Xiurong, Li Guangwu. Microbiology Experiment (third edition). Beijing: Higher Education Publishing House, 1999.) measure the physiological and biochemical property that above-mentioned Acetochlor/weeds burn degradation bacteria 198-r-47.
The physiological and biochemical property measurement result that described Acetochlor/weeds burn degradation bacteria 198-r-47 is as shown in table 2:
Table 2. Acetochlors/weeds burn the physiological and biochemical property of degradation bacteria 198-r-47
Note: "+" representing positive, "-" represents negative.
3rd, 16s rdna sequence homology analysis
Conventional method culture above-mentioned steps one isolate and purify the Acetochlor/weeds obtaining and burn degradation bacteria 198-r-47, extract Total dna of bacterial strain as gene magnification template, with bacterium 16s rdna universal primer, 27f:5 '- Agagtttgatcctggctcag-3 ', 1492r:5 '-taccttgttacgactt-3 ' carries out pcr reaction.Reaction system adopts Shanghai bioengineering Co., Ltd pcr amplification kit.Response procedures are: 95 DEG C of denaturation 30s, 55 DEG C of annealing 1min, 72 DEG C prolong Stretch 2min, totally 30 circulations.Dna sequencing is won polygala root biotech company by Beijing three and is completed, sequence assembly and similarity analysis Completed using dnastar software, gene compare by American National Biotechnology Information center ncbi database (http: // Www.ncbi.nlm.nih.gov) complete online with eztaxon.
Pcr gene magnification obtains the 16s rdna genetic fragment about 1.3kb that Acetochlor/weeds burn degradation bacteria 198-r-47, Carry out online sequence analysis, result with 16s rdna sequence published in ncbi and eztaxon database after measuring sequence Display Acetochlor/weeds burn degradation bacteria 198-r-47 and rhizobium rhizobium pusense nrcpb10tHomology is Height, reaches 99.31%.
The 16s rdna sequence that Acetochlor/weeds burn degradation bacteria 198-r-47 refers to sequence 1.
In view of above-mentioned form, analysis of physio biochemical characteristics and 16s rdna sequence homology analysis result, by step one point Acetochlor/the weeds obtaining from purifying are burnt degradation bacteria 198-r-47 and are accredited as rhizobium (rhizobium sp.).This second grass Amine/weeds burn degradation bacteria 198-r-47, and to be preserved in China Committee for Culture Collection of Microorganisms on October 28th, 2014 general Logical microorganism center (abbreviation cgmcc, address is: Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3), deposit number is cgmcc no.9867.The full name that Acetochlor/weeds burn degradation bacteria 198-r-47 is rhizobium (rhizobium sp.) 198-r-47 Cgmcc no.9867, referred to as rhizobium (rhizobium sp.) 198-r-47.
Embodiment 2, rhizobium (rhizobium sp.) 198-r-47 cgmcc no.9867 degraded Acetochlor ability are quantitative Measure
First, Acetochlor bioassay standard curve plotting
Accurately weigh Acetochlor standard items (fluka Products) 10.0mg in 10ml volumetric flask, molten with a small amount of methyl alcohol Solution, volumetric flask is placed on vibration 10min in ultrasonic wave bath, then with methanol constant volume to 10ml, shakes up, be made into 1000mg/l second Careless amine mother liquor.Then with methanol dilution obtain concentration be respectively 10,20,40,60,80,100mg/l standard liquid, using efficient Liquid chromatography (hplc) measures the peak area of variable concentrations Acetochlor standard items, 3 repetitions.With the concentration of Acetochlor for horizontal seat Mark, peak area is ordinate, draws Acetochlor calibration curve, as shown in Figure 1.
Testing conditions are as follows:
Detecting system: agilent 1100series.Chromatographic column: c18diamosiltmReversed-phase column, 250mm × 4.6mm, grain 5 μm of footpath.Chromatographic condition: mobile phase: methyl alcohol: water=80:20 (v/v), water glacial acetic acid is adjusted to ph=3；Detection wavelength, 215nm； Flow velocity, 1.0ml/min；Sampling volume, 20 μ l；30 DEG C of column temperature.
Gained Acetochlor bioassay standard curve: y=55.363x+144.5 (r2=0.9999).Wherein, y is peak area, x For Acetochlor concentration.
2nd, rhizobium (rhizobium sp.) 198-r-47 cgmcc no.9867 degraded Acetochlor ability quantitative determination
Rhizobium (rhizobium sp.) 198-r-47 process: by rhizobium (rhizobium sp.) 198-r-47 Cgmcc no.9867 is seeded in culture 24h on beef extract-peptone solid medium, and picking 1 ring is seeded in 5ml beef extract albumen In peptone fluid nutrient medium, 180r/min cultivates 12h, and centrifugation is removed culture medium, is adjusted to od with inorganic salt liquid culture medium600Value For 1.0.Draw 0.2ml bacteria suspension (1 × 109Cfu/ml) it is inoculated into the inorganic salt liquid culture medium that 5ml contains Acetochlor 50mg/l (Acetochlor (fluka Products) is added to make the training that the concentration of Acetochlor obtains for 50mg/l in inorganic salt liquid culture medium Foster base) test tube in, 28 DEG C, 180r/min cultivate 7 days, obtain degradation solution, as follows measure Residual Determination of Acetochlor: take To 50ml centrifuge tube, 8000r/min centrifugation 5min collects supernatant to 3ml degradation solution, adds 3ml dichloromethane, acutely vibrates 5min, stands 10min, treats aqueous phase and organic phase layering, adds a small amount of anhydrous sodium sulfate that organic phase is dehydrated, accurately draw 800 μ l organic phases, in the centrifuge tube of 1.5ml, are dried up with Nitrogen evaporator, add 400 μ l methyl alcohol, ultrasonic in ultrasonic cleaner 10min, is filtered to liquid chromatogram sample bottle with 0.22 μm of organic phase aculeus type filter, measures second grass according to above-mentioned hplc method Amine.
Rhizobium (rhizobium sp.) ld1616 process: except by above-mentioned rhizobium (rhizobium sp.) 198-r-47 Rhizobium (rhizobium sp.) 198-r-47 cgmcc no.9867 in process replaces with rhizobium (rhizobium Sp.) ld1616cgmcc no.7775, remaining all same.
Blank is processed: meanwhile, with non-Rhizobium Inoculation (rhizobium sp.) 198-r-47 cgmcc no.9867 The above-mentioned minimal medium containing Acetochlor 50mg/l as blank, measure the concentration of Acetochlor according to the method described above.
Acetochlor degradation rate: x=(1-a/b) × 100%, in formula, x is degradation rate (%), and a is rhizobium (rhizobium Sp.) remain in the Acetochlor concentration of residual or rhizobium (rhizobium sp.) ld1616 treatment fluid in 198-r-47 treatment fluid Acetochlor concentration, b is the Acetochlor concentration not connecing in bacterium blank treatment fluid residual.Experiment sets 3 repetitions, repeats every time Each processes 10 test tubes of inoculation.
Table 3. rhizobium (rhizobium sp.) 198-r-47 cgmcc no.9867 degraded Acetochlor effect
Result shows, Acetochlor initial concentration is 50mg/l, described rhizobium (rhizobium sp.) 198-r- after 7 days 47 cgmcc no.9867 reach 73.65% (as shown in table 3) to the degradation rate of Acetochlor；Rhizobium (rhizobium sp.) Ld1616 reaches 0% to the degradation rate of Acetochlor.This result shows, rhizobium (rhizobium provided by the present invention Sp.) the degradable Acetochlor of 198-r-47 cgmcc no.9867.
Embodiment 3, rhizobium (rhizobium sp.) 198-r-47 cgmcc no.9867 degraded weeds burn ability quantitation Measure
First, weeds burn the drafting of bioassay standard curve
Accurately weighing weeds, to burn standard items (fluka Products) 10.0mg in 10ml volumetric flask, molten with a small amount of methyl alcohol Solution, volumetric flask is placed on vibration 10min in ultrasonic wave bath, then with methanol constant volume to 10ml, shakes up, be made into 1000mg/l miscellaneous Grass burns mother liquor.Then with methanol dilution obtain concentration be respectively 10,20,40,60,80,100mg/l standard liquid.Using efficient Liquid chromatogram (hplc) measures the peak area that variable concentrations weeds burn standard items, 3 repetitions.Burn concentration with weeds as abscissa, Peak area is ordinate, draws weeds and burns calibration curve, as shown in Figure 2.
Testing conditions are as follows:
Detecting system: agilent 1100series.Chromatographic column: c18diamosiltmReversed-phase column, 250mm × 4.6mm, grain 5 μm of footpath.Chromatographic condition: mobile phase: acetonitrile: water (glacial acetic acid adjusts ph3.0)=40:60 (v/v)；Detection wavelength, 258nm；Flow velocity, 1.0ml/min；Sampling volume, 10 μ l；30 DEG C of column temperature.
Gained weeds burn bioassay standard curve: y=7.3196x+2.1212 (r2=0.9875).Wherein, y is peak area, x Burn concentration for weeds.
2nd, rhizobium (rhizobium sp.) 198-r-47 cgmcc no.9867 degraded weeds burn ability quantitative determination
Rhizobium (rhizobium sp.) 198-r-47 process: accurately add 5ml to burn 100mg/l containing weeds in test tube Minimal medium (in inorganic salt liquid culture medium add weeds burn the training making the concentration that weeds are burnt obtain for 100mg/l Foster base), add the od of 0.2ml600It is worth rhizobium (rhizobium sp.) the 198-r-47 cgmccno.9867 bacterium for 1.0 Liquid (1 × 109Cfu/ml), 28 DEG C, 200r/min cultivate 7 days, obtain degradation solution.Take 4ml degradation solution to 50ml centrifuge tube, plus Enter 4ml acetonitrile, vibrate 2min, stand 10min, add a little anhydrous sodium sulfate.Accurately draw 800 μ l organic phases to be transferred to In 1.5ml ep centrifuge tube, Nitrogen evaporator dries up, add 400 μ l methyl alcohol (chromatographic grade), so that weeds is burnt under ul-trasonic irradiation It is completely dissolved, be collected by filtration to sample bottle with liquid spectrum filter, measure weeds according to above-mentioned hplc method and burn.
Rhizobium (rhizobium sp.) ld1616 process: except by above-mentioned rhizobium (rhizobium sp.) 198-r-47 Rhizobium (rhizobium sp.) 198-r-47 cgmcc no.9867 in process replaces with rhizobium (rhizobium Sp.) ld1616cgmcc no.7775, remaining all same.
Blank is processed: meanwhile, with non-Rhizobium Inoculation (rhizobium sp.) 198-r-47 cgmcc no.9867 Above-mentioned burn the inorganic salt liquid culture medium of 100mg/l as blank containing weeds, measure what weeds were burnt according to the method described above Concentration.
Weeds burn degradation rate: x=(1-a/b) × 100%, and in formula, x is degradation rate (%), and a is rhizobium (rhizobium Sp.) in 198-r-47 treatment fluid, the weeds of residual burn residual in concentration or rhizobium (rhizobium sp.) ld1616 treatment fluid Weeds burn concentration, b is that the weeds not connecing in bacterium blank treatment fluid residual burn concentration.Experiment sets 3 repetitions, repeats every time Each processes 10 test tubes of inoculation.Measurement result is as shown in table 4.
Result shows, it is 100mg/l, described rhizobium (rhizobium sp.) 198-r- after 7 days that weeds burn initial concentration The degradation rate 9.04% that 47 cgmcc no.9867 burn to weeds；Rhizobium (rhizobium sp.) ld1616 burns to weeds Degradation rate is 0.02% (as shown in table 4).This result shows, rhizobium (rhizobium sp.) provided by the present invention The degradable weeds of 198-r-47 cgmcc no.9867 are burnt.
Table 4. rhizobium (rhizobium sp.) 198-r-47 cgmcc no.9867 degraded weeds burn effect

Claims (4)

1. application in degraded Acetochlor and/or weeds burn for rhizobium (rhizobium sp.) 198-r-47, described rhizobium (rhizobium sp.) 198-r-47 is in the deposit number of China Committee for Culture Collection of Microorganisms's common micro-organisms center For cgmcc no.9867.
2. application in degraded Acetochlor and/or weeds burn for the microbial inoculum, the active component of described microbial inoculum is rhizobium (rhizobium sp.) 198-r-47, described rhizobium (rhizobium sp.) 198-r-47 is in Chinese microorganism strain preservation The deposit number of administration committee's common micro-organisms center is cgmcc no.9867.
3. rhizobium (rhizobium sp.) 198-r-47 burns the application in pollution, institute in rehabilitating soil Acetochlor and/or weeds State rhizobium (rhizobium sp.) 198-r-47 in China Committee for Culture Collection of Microorganisms's common micro-organisms center Deposit number is cgmcc no.9867.
4. microbial inoculum burns the application in pollution in rehabilitating soil Acetochlor and/or weeds, and the active component of described microbial inoculum is rhizobium (rhizobium sp.) 198-r-47, described rhizobium (rhizobium sp.) 198-r-47 is in Chinese microorganism strain preservation The deposit number of administration committee's common micro-organisms center is cgmcc no.9867.
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