CN106424125A - Microbial agent for treating heavy metal polluted soil, preparation method and applications thereof - Google Patents

Microbial agent for treating heavy metal polluted soil, preparation method and applications thereof Download PDF

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Publication number
CN106424125A
CN106424125A CN201610570018.4A CN201610570018A CN106424125A CN 106424125 A CN106424125 A CN 106424125A CN 201610570018 A CN201610570018 A CN 201610570018A CN 106424125 A CN106424125 A CN 106424125A
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Prior art keywords
trichoderma asperellum
soil
mortierella alpina
heavy metal
mucor
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CN106424125B (en
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张旭
杨欢欢
崔兆杰
孙立群
张萌萌
魏绍山
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Shandong University
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Shandong University
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    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B09—DISPOSAL OF SOLID WASTE; RECLAMATION OF CONTAMINATED SOIL
    • B09C—RECLAMATION OF CONTAMINATED SOIL
    • B09C1/00—Reclamation of contaminated soil
    • B09C1/10—Reclamation of contaminated soil microbiologically, biologically or by using enzymes
    • B09C1/105—Reclamation of contaminated soil microbiologically, biologically or by using enzymes using fungi or plants
    • 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
    • C12N1/14—Fungi; Culture media therefor

Abstract

The present invention relates to a microbial agent for treating heavy metal polluted soil, a preparation method and applications thereof. The microbial agent comprises the following raw materials: rice husk, pig manure, chicken manure, soybean powder, single cell protein, a Mortierella alpina fermentation broth, a Trichoderma asperellum fermentation broth, and a Mucor circinelloides fermentation broth. According to the present invention, the microbial agent prepared by mixing Mortierella alpina, Trichoderma asperellum and Mucor circinelloides according to a certain ratio is firstly used, can be used in the complex and changing environment, and can provide effects in the poor soil environment, wherein the functional microbes provide high tolerance to heavy metals, and can maintain activity and perform efficient repair in the heavily heavy metal polluted soil.

Description

A kind of microbial bacterial agent processing heavy metal pollution of soil and preparation method and application
Technical field
The present invention relates to a kind of microbial bacterial agent processing heavy metal pollution of soil and preparation method and application, belong to soil Earth Treatment process field.
Background technology
Ecological disruption problem has become the key issue of restriction human development.Refuse dump that mining activities is formed, mine tailing Storehouse and ambient contamination place often pollutant component are complicated, containing heavy metal and Some Organic Pollutants, the dirt of these heavy metals Dye concentration is high, pollution level weight, causes animals and plants to be difficult to grow wherein, has a strong impact on ecological environment.Heavy metal in soil, Can pass through surface water body runoff, air floating dust, pollute soil around, waters and air, it affect face will be considerably beyond giving up The region of gurry storeyard and space, pollution effect to spend a large amount of human and material resources, financial resources through being lot more time to recover, And be difficult to return to original level.
The soil restoring technology of modern technologies reply heavy metal pollution is relatively deficient, is concentrated mainly on hyperaccumulative plant to spy Determining the enrichment of heavy metal, the method can only tackle more single heavy metal pollution, and plant is to the requirement of soil environment relatively Height, limits its range of application.
Chinese patent literature CN104070062A (application number 201310108243.2) discloses a kind of heavy metal polluted soil The microorganism in situ restorative procedure of earth, its step is：(1) bacteria group culture：(2) flora activates；(3) growth of microorganism agent preparation；(4) seek Nutrient solution is prepared；(5) just throw；(6) repair.By a huge sum of money in the microorganism in situ restorative procedure of the heavy-metal contaminated soil that this invention provides Microorganism belonging to genus degradation flora, growth of microorganism accelerator as main component, supplement again certain microbial nutrition element, to While soil puts into preferred heavy metal microorganism conversion flora, increase growth of microorganism accelerator and promote its breeding at a high speed, Thus greatly improve conversion and the degradation efficiency of heavy metals in soil, solve heavy-metal contaminated soil microorganism in situ Time length in restorative procedure, take effect slow problem, makes microorganism in situ recovery technique become reliable, efficient, exercisable heavy Metallic pollution soil remediation means.
How to find the toxicity of the heavy metal that can quickly effectively reduce in polluted soil, become current research heat Point.
Content of the invention
The present invention is directed to the deficiencies in the prior art, a kind of microbial bacterial agent processing heavy metal pollution of soil and system thereof are provided Preparation Method and application.
Technical solution of the present invention is as follows：
A kind of microbial bacterial agent processing heavy metal pollution of soil, is made up of the raw material of following weight portion：
50～80 parts of rice husk, pig manure 20～50 parts, chicken manure 20～50 parts, soy meal 30～60 parts, single cell protein 4～6 Part, Mortierella alpina (Mortierella alpina) zymotic fluid 2.5-3.5 part, trichoderma asperellum (Trichoderma Asperellum) zymotic fluid 1.5～2.5 parts, roll up branch Mucor (Mucor circinelloides) zymotic fluid 0.5～1.5 part；
Cell concentration in described Mortierella alpina (Mortierella alpina) zymotic fluid is 0.4 × 108cfu/ml； Cell concentration in trichoderma asperellum (Trichoderma asperellum) zymotic fluid is 1.0 × 108cfu/ml；Volume branch Mucor Cell concentration in (Mucor circinelloides) zymotic fluid is 3.6 × 108cfu/ml；In microbial bacterial agent, thalline always contains Amount is 6.8 × 108cfu/g.
According to currently preferred, described Mortierella alpina (Mortierella alpina), trichoderma asperellum (Trichoderma asperellum) is 1 with volume branch Mucor (Mucor circinelloides) viable bacteria body ratio of number： (1.0～2.5):(1.8～3.6).
According to the present invention it is further preferred that described Mortierella alpina (Mortierella alpina), trichoderma asperellum (Trichoderma asperellum) is 1 with volume branch Mucor (Mucor circinelloides) viable bacteria body ratio of number： 1.5：3.
According to currently preferred, described thalline total content is 6.8 × 108cfu/g.
According to currently preferred, described Mortierella alpina (Mortierella alpina) is Mortierella alpina (Mortierella alpina) ME-AA01, derives from China typical culture collection center (CCTCC), bacterium numbering CCTCC NO:M207067.
According to currently preferred, described trichoderma asperellum (Trichoderma asperellum) is trichoderma asperellum (Trichoderma asperellum) PD-19, derives from China typical culture collection center (CCTCC), bacterium numbering CCTCC NO:M2013113.
According to currently preferred, described volume branch Mucor (Mucor circinelloides) derives from Holland's microbial bacteria Plant preservation center (CBS), bacterium numbering CBS131818.
The preparation method of the microbial bacterial agent of above-mentioned process heavy metal pollution of soil, step is as follows：
(1) Mortierella alpina (Mortierella alpina) is cultivated through first order seed cultivation, secondary seed and expands Cultivate, prepare 1.2 × 107～1.6 × 107Mortierella alpina (Mortierella alpina) zymotic fluid of cfu/ml；
(2) trichoderma asperellum (Trichoderma asperellum) is cultivated through first order seed cultivation, secondary seed and expands Big cultivation, prepares 0.6～0.8 × 108Trichoderma asperellum (Trichoderma asperellum) zymotic fluid of cfu/ml；
(3) volume branch Mucor (Mucor circinelloides) is cultivated through first order seed cultivation, secondary seed and expands Cultivate, prepare 1.4 × 108～1.8 × 108Volume branch Mucor (Mucor circinelloides) zymotic fluid of cfu/ml；
(4) Mortierella alpina (Mortierella alpina) zymotic fluid, the step (2) preparing step (1) prepares The volume branch Mucor (Mucor that trichoderma asperellum (Trichoderma asperellum) zymotic fluid, step (3) prepare Circinelloides) zymotic fluid and rice husk, pig manure, chicken manure, soy meal and single cell protein mix, and room temperature condition is transferred Put 4～6 weeks, stir weekly 2～4 times, be dried and be less than 20% to moisture content, prepare microbial bacterial agent.
According to currently preferred, in described step (1), (2) and step (3), first order seed condition of culture is：Illumination 1000～3000LX, cultivation temperature 25～30 DEG C, Anaerobic culturel 3～4 days；
Every liter of component of described primary-seed medium is as follows：
Glucose 15～20g, peptone 6～10g, yeast extract 1～3g, dipotassium hydrogen phosphate 0.2～0.8g, magnesium sulfate 0.01～0.03g, calcium chloride 0.1～0.29g.
According to currently preferred, in described step (1), (2) and step (3), secondary seed condition of culture is：Illumination 1000～3000LX, cultivation temperature 25～30 DEG C, Anaerobic culturel 3～4 days；
Every liter of component of described secondary seed medium is as follows：
Glucose 15～20g, peptone 6～10g, yeast extract 1～3g, dipotassium hydrogen phosphate 0.2～0.8g, magnesium sulfate 0.01～0.03g, calcium chloride 0.1～0.29g.
According to currently preferred, in described step (1), (2) and step (3), expanding condition of culture is：Cultivation temperature 28 ～30 DEG C, dark aerobic cultivate 6～8 days, ratio of ventilating per hour is liquid and gas volume ratio 1:0.5, speed of agitator 200～ 300r/min；
The culture medium every liter component that described expansion is cultivated is as follows：
Glucose 15～20g, peptone 6～10g, yeast extract 1～3g, dipotassium hydrogen phosphate 0.2～0.8g, magnesium sulfate 0.01～0.03g, calcium chloride 0.1～0.29g.
The application of the microbial bacterial agent of above-mentioned process heavy metal pollution of soil, step is as follows：
The microbial bacterial agent processing heavy metal pollution of soil is directly applied in the soil of heavy metal pollution as base manure, Turning over mixing, amount of application is 8～12g/ square metre, processes 33～38 days under conditions of temperature 29～31 DEG C, pH7.5～8.5.
Above-mentioned rice husk, pig manure, chicken manure, soy meal and single cell protein etc. are common commercially available prod, this area.
Beneficial effect
1st, the present invention uses Mortierella alpina (Mortierella alpina), trichoderma asperellum (Trichoderma first Asperellum) the process heavy metal pollution of soil being mixed to prepare in proportion with volume branch Mucor (Mucor circinelloides) Microbial bacterial agent, this microbial inoculum can be applied among environment complicated and changeable, in the face of poor soil environment, still can send out Waving effect, the tolerance of functional microorganism heavy metal is high, remains to keep activity simultaneously in the soil of heavy metal serious pollution Efficiently repair；
2nd, the microbial bacterial agent preparation technology of process heavy metal pollution of soil of the present invention is simple, applied widely, becomes This is low, workable.The repair mechanisms of functional microorganism heavy metal are various, can not only heavy metal absorption accumulate, moreover it is possible to Enough heavy metal is converted into stable state, administers heavy metal-polluted on the basis of not causing secondary to destroy environment to greatest extent Dye, the strong adaptability to environment for the functional microorganism in composite bacteria agent, thus considerably increase the range of application of composite bacteria agent.
Detailed description of the invention
Below in conjunction with embodiment, technical scheme is described further, but institute of the present invention protection domain is not limited to This.
Biological material source：
Mortierella alpina (Mortierella alpina) is Mortierella alpina (Mortierella alpina) ME- AA01, derives from China typical culture collection center (CCTCC), bacterium numbering CCTCC NO:M207067；
Trichoderma asperellum (Trichoderma asperellum) is trichoderma asperellum (Trichoderma asperellum) PD- 19, derive from China typical culture collection center (CCTCC), bacterium numbering CCTCC NO:M2013113；
Volume branch Mucor (Mucor circinelloides) derives from Holland's Culture Collection (CBS), bacterial classification Numbering CBS131818.
Rice husk described in embodiment, pig manure, chicken manure, soy meal and single cell protein are common commercially available prod, this area.
Embodiment 1
A kind of microbial bacterial agent processing heavy metal pollution of soil, is made up of the raw material of following weight portion：
80 parts of rice husk, pig manure 20 parts, chicken manure 50 parts, soy meal 30 parts, single cell protein 6 parts, Mortierella alpina (Mortierella alpina) zymotic fluid 2.5 parts, trichoderma asperellum (Trichoderma asperellum) zymotic fluid 2.5 parts, Volume branch Mucor (Mucor circinelloides) zymotic fluid 0.5 part；
In microbial bacterial agent, thalline total content is 5.3 × 108cfu/g；Described Mortierella alpina (Mortierella Alpina), trichoderma asperellum (Trichoderma asperellum) and volume branch Mucor (Mucor circinelloides) viable bacteria Body ratio of number is 1：1.8:2.5.
The preparation method of the microbial bacterial agent of above-mentioned process heavy metal pollution of soil, step is as follows：
(1) Mortierella alpina (Mortierella alpina) is cultivated through first order seed cultivation, secondary seed and expands Cultivate, prepare 0.4 × 108Mortierella alpina (Mortierella alpina) zymotic fluid of cfu/ml；
(2) trichoderma asperellum (Trichoderma asperellum) is cultivated through first order seed cultivation, secondary seed and expands Big cultivation, prepares 1.0 × 108Trichoderma asperellum (Trichoderma asperellum) zymotic fluid of cfu/ml；
(3) volume branch Mucor (Mucor circinelloides) is cultivated through first order seed cultivation, secondary seed and expands Cultivate, prepare 3.6 × 108Volume branch Mucor (Mucor circinelloides) zymotic fluid of cfu/ml；
(4) Mortierella alpina (Mortierella alpina) zymotic fluid, the step (2) preparing step (1) prepares The volume branch Mucor (Mucor that trichoderma asperellum (Trichoderma asperellum) zymotic fluid, step (3) prepare Circinelloides) zymotic fluid and rice husk, pig manure, chicken manure, soy meal and single cell protein mix, and room temperature condition is transferred Put 4 weeks, stir weekly 4 times, be dried and be less than 20% to moisture content, prepare microbial bacterial agent.
In described step (1), (2) and step (3), first order seed condition of culture is：Illumination 2000LX, cultivation temperature 27 DEG C, Anaerobic culturel 3 days；Every liter of component of described primary-seed medium is as follows：
Glucose 15g, peptone 10g, yeast extract 1g, dipotassium hydrogen phosphate 0.8g, magnesium sulfate 0.01g, calcium chloride 0.29g.
In described step (1), (2) and step (3), secondary seed condition of culture is：Illumination 2000LX, cultivation temperature 27 DEG C, Anaerobic culturel 3 days；Every liter of component of described secondary seed medium is as follows：
Glucose 15g, peptone 10g, yeast extract 1g, dipotassium hydrogen phosphate 0.8g, magnesium sulfate 0.01g, calcium chloride 0.29g.
In described step (1), (2) and step (3), expanding condition of culture is：Cultivation temperature 28 DEG C, dark aerobic cultivation 8 My god, ratio of ventilating per hour is liquid and gas volume ratio 1:0.5, speed of agitator 300r/min；
The culture medium every liter component that described expansion is cultivated is as follows：
Glucose 15g, peptone 10g, yeast extract 1g, dipotassium hydrogen phosphate 0.8g, magnesium sulfate 0.01g, calcium chloride 0.29g.
Embodiment 2
A kind of microbial bacterial agent processing heavy metal pollution of soil, is made up of the raw material of following weight portion：
50 parts of rice husk, pig manure 50 parts, chicken manure 20 parts, soy meal 60 parts, single cell protein 4 parts, Mortierella alpina (Mortierella alpina) zymotic fluid 3.5 parts, trichoderma asperellum (Trichoderma asperellum) zymotic fluid 1.5 parts, Volume branch Mucor (Mucor circinelloides) zymotic fluid 1.5 parts；
In microbial bacterial agent, thalline total content is 8.3 × 108cfu/g；Described Mortierella alpina (Mortierella Alpina), trichoderma asperellum (Trichoderma asperellum) and volume branch Mucor (Mucor circinelloides) viable bacteria Body ratio of number is 1：1:3.6.
Preparation method is with embodiment 1.
Embodiment 3
A kind of microbial bacterial agent processing heavy metal pollution of soil, is made up of the raw material of following weight portion：
65 parts of rice husk, pig manure 35 parts, chicken manure 35 parts, soy meal 45 parts, single cell protein 5 parts, Mortierella alpina (Mortierella alpina) zymotic fluid 3 parts, trichoderma asperellum (Trichoderma asperellum) zymotic fluid 2 parts, roll up branch Mucor (Mucor circinelloides) zymotic fluid 1 part；
In microbial bacterial agent, thalline total content is 6.8 × 108cfu/g；Described Mortierella alpina (Mortierella Alpina), trichoderma asperellum (Trichoderma asperellum) and volume branch Mucor (Mucor circinelloides) viable bacteria Body ratio of number is 1:1.5:3.
Preparation method is with embodiment 1.
Comparative example 1
The system of the microbial bacterial agent as described in Chinese patent literature CN104070062A (application number 201310108243.2) Preparation Method, step is as follows：
(i) bacteria group culture：Selection can convert the microbial degradation flora body of heavy metal characteristic, uses the side of solid culture Formula, carries out cultivating domestication with wheat bran for carrier, the powdered substance being formed after taking the mixing of its middle bacterial product mummification；
(ii) flora activates：Powdered substance in step (i) is put in water, soaks 0.5～2h, remove wheat bran and get final product Flora liquid after activation；
(iii) growth of microorganism agent preparation：Compound amino acid, composite trace element, activating enzymes and humic acid are put into water In, stir, obtain growth of microorganism agent；
(iv) compounding：Growth of microorganism agent adds microbial nutrition element, adds water and diluted, obtain microorganism Growth stimulator nutrient solution；
Comparative example 2
Processing the microbial bacterial agent of heavy metal pollution of soil as described in Example 3, difference is,
Trichoderma asperellum replaces with trichoderma reesei (Trichoderma reesei), and this bacterium source is in the common micro-life of China Thing culture presevation administrative center, bacterium numbering CGMCC No：9644.
Comparative example 3
Processing the microbial bacterial agent of heavy metal pollution of soil as described in Example 3, difference is,
Volume branch Mucor replaces with Mucor racemosus (Mucor racemosus), and this bacterium source is in Chinese Typical Representative culture Preservation center, bacterium numbering CCTCC NO：M201026.
Experimental example 1
Soil moved in improve the original by 2/3 and 1/3 Nutrition Soil mixing after, respectively add heavy metal contaminants background value the 10th, the 100th, the 500th, After 1000 times, analog composite heavy-metal contaminated soil, balance 2-3 days.Apply respectively described in embodiment 1-3 and comparative example 1-3 Composite bacteria agent, amount of application is 6kg/ mu, is spaced 7 days and applies once after mixing of turning over, continuous administration 5 times.Every seven days observing earths The physicochemical property of the change of content of beary metal and soil in earth.After applying composite bacteria agent, heavy metal in soil content significantly reduces. Result is as shown in the table：
Table 1
After soil improvement, plant black nightshade, summer cypress, the fluffy three kinds of plants of alkali, every physical signs of periodic monitoring plant, plant Grow fine.This composite bacteria agent can not only administer heavy metal pollution effectively, and can promote the growth of plant.
Experimental example 2
Yellow triangle has Unutilized Energy about 8,100,000 mu, and wherein salt-soda soil just account for 2,700,000 mu, northern in Dongying Lijin, Northeast, Kenli and extensively also have the soil on the ground such as Hekou District all by salinization of soil around east.Dongying salt-soda soil not only soil environment is disliked Bad, and there is more serious heavy metal pollution.By surface water body runoff, air floating dust, pollute soil around, waters And air, surrounding environment is caused and has a strong impact on.Apply the composite bacteria agent described in embodiment 1-3 and comparative example 1-3, administration respectively Amount is 7kg/ mu, is spaced 7 days and applies once after mixing of turning over, continuous administration 5 times.After applying composite bacteria agent, the gas permeability of soil Dramatically increasing, the soil weight is from 1.49g/m3Drop to 1.24g/m3, reached normal Soil standard (1.1-1.4g/m3).Soil Water-retaining property aobvious improve, the water content of soil brings up to 21% from 15%.Comparative example does not has significant change.A huge sum of money in soil Belonging to content to significantly reduce, detecting heavy metal removing rate when the 30th day, result is as described in Table 2：
Table 2
Result shows, after applying this composite bacteria agent, microorganism can adapt to alkaline environment, keeps good survival state. After administration composite bacteria agent, in mine tailing soil, content of beary metal significantly reduces, and the physical and chemical parameter of soil all obtains certain raising.
Experimental example 3
Discarded object (mine tailing peels off rock etc.) during mining needs large-area place of banking up, in city, Anshan City's periphery defines occupation of land 7,910,000 m2Discarded refuse dump and 2,520,000 m2Discarded tailing storehouse.Cause taking the excess in soil Destruction with the ecosystem original to storeyard；It affects face by considerably beyond the region of discarded object storeyard and space.Mine tailing District's soil is alkalescence, soil depletion, poor water retention property, and heavy metal pollution is more serious, it is difficult to repair.Apply reality in mine tailing district respectively Executing the composite bacteria agent described in example 1-3 and comparative example 1-3, amount of application is 8kg/ mu, is spaced 7 days and applies once, even after mixing of turning over Continuous administration 5 times.Heavy metal removing rate is detected when the 30th day.After applying composite bacteria agent, the gas permeability of soil dramatically increases, and soil holds Heavy from 1.57g/m3Drop to 1.33g/m3, reached normal Soil standard (1.1-1.4g/m3).The water-retaining property of soil is aobvious to be improved, The water content of soil brings up to 18% from 9%.This index does not has significant change in comparative example.Heavy metal in soil content is notable Reducing, detecting heavy metal removing rate when the 30th day, result is as described in Table 3：
Table 3
Result shows, after applying this composite bacteria agent, microbe survival in order, can adapt to rugged environment.Administration After composite bacteria agent, in mine tailing soil, content of beary metal significantly reduces.This composite bacteria agent heavy metal contaminated soil has preferably repaiies Multiple effect.
Interpretation of result
By the contrast discovery of embodiment, this composite bacteria agent has preferable removal effect to five heavy metal species.High mountain quilt Spore mould (Mortierella alpina), trichoderma asperellum (Trichoderma asperellum) and volume branch Mucor (Mucor Circinelloides) optimal proportion is 1:1.5:3.
By comparative example it is found that Mortierella alpina (Mortierella alpina) in this patent, trichoderma asperellum There is between (Trichoderma asperellum) and volume branch Mucor (Mucor circinelloides) mutually collaborative work With the clearance of heavy metal can be significantly improved.
Described only presently preferred embodiments of the present invention, it is impossible to this implementation condition limiting the present invention and practical range.Depend on Modification that scope of the present invention patent and invention description content are made and change, still belong to the scope that patent of the present invention covers.

Claims (10)

1. the microbial bacterial agent processing heavy metal pollution of soil, it is characterised in that be made up of the raw material of following weight portion：
50～80 parts of rice husk, pig manure 20～50 parts, chicken manure 20～50 parts, soy meal 30～60 parts, single cell protein 4～6 parts are high Mountain Mortierella (Mortierella alpina) zymotic fluid 3～5 parts, trichoderma asperellum (Trichoderma asperellum) ferments Liquid 1～3 part, rolls up branch Mucor (Mucor circinelloides) zymotic fluid 0.5～1.5 part；
Cell concentration in described Mortierella alpina (Mortierella alpina) zymotic fluid is 0.4 × 108cfu/ml；Spine spore Cell concentration in wood mould (Trichoderma asperellum) zymotic fluid is 1.0 × 108cfu/ml；Volume branch Mucor (Mucor Circinelloides) cell concentration in zymotic fluid is 3.6 × 108cfu/ml；In microbial bacterial agent, thalline total content is 6.8 ×108cfu/g.
2. the microbial bacterial agent processing heavy metal pollution of soil as claimed in claim 1, it is characterised in that described high mountain quilt Spore mould (Mortierella alpina), trichoderma asperellum (Trichoderma asperellum) and volume branch Mucor (Mucor Circinelloides) viable bacteria body ratio of number is 1：(1.0～2.5):(1.8～3.6).
3. the microbial bacterial agent processing heavy metal pollution of soil as claimed in claim 2, it is characterised in that described high mountain quilt Spore mould (Mortierella alpina), trichoderma asperellum (Trichoderma asperellum) and volume branch Mucor (Mucor Circinelloides) viable bacteria body ratio of number is 1:1.5:3.
4. the microbial bacterial agent processing heavy metal pollution of soil as claimed in claim 1, it is characterised in that described thalline always contains Amount is 6.8 × 108cfu/g.
5. the microbial bacterial agent processing heavy metal pollution of soil as claimed in claim 1, it is characterised in that described Mortierella alpine Mould (Mortierella alpina) is Mortierella alpina (Mortierella alpina) ME-AA01, derives from Chinese Typical Representative Culture collection center (CCTCC), bacterium numbering CCTCC NO:M207067；
Preferably, described trichoderma asperellum (Trichoderma asperellum) is trichoderma asperellum (Trichoderma Asperellum) PD-19, derives from China typical culture collection center (CCTCC), bacterium numbering CCTCC NO: M2013113；
Preferably, described volume branch Mucor (Mucor circinelloides) derives from Holland's Culture Collection (CBS), bacterium numbering CBS131818.
6. process the preparation method of the microbial bacterial agent of heavy metal pollution of soil described in claim 1, it is characterised in that step is such as Under：
(1) Mortierella alpina (Mortierella alpina) is cultivated and is expanded cultivate through first order seed cultivation, secondary seed, Prepare 0.4 × 108Mortierella alpina (Mortierella alpina) zymotic fluid of cfu/ml；
(2) trichoderma asperellum (Trichoderma asperellum) is cultivated through first order seed cultivation, secondary seed and expands training Support, prepare 1.0 × 108Trichoderma asperellum (Trichoderma asperellum) zymotic fluid of cfu/ml；
(3) volume branch Mucor (Mucor circinelloides) is cultivated and is expanded cultivate through first order seed cultivation, secondary seed, Prepare 3.6 × 108Volume branch Mucor (Mucor circinelloides) zymotic fluid of cfu/ml；
(4) the spine spore that Mortierella alpina (Mortierella alpina) zymotic fluid prepared for step (1), step (2) are prepared The volume branch Mucor (Mucor that wood mould (Trichoderma asperellum) zymotic fluid, step (3) prepare Circinelloides) zymotic fluid and rice husk, pig manure, chicken manure, soy meal and single cell protein mix, and room temperature condition is transferred Put 4～6 weeks, stir weekly 2～4 times, be dried and be less than 20% to moisture content, prepare microbial bacterial agent.
7. preparation method as claimed in claim 6, it is characterised in that in described step (1), (2) and step (3), one-level kind Sub-condition of culture is：Illumination 1000～3000LX, cultivation temperature 25～30 DEG C, Anaerobic culturel 3～4 days；
Every liter of component of described primary-seed medium is as follows：
Glucose 15～20g, peptone 6～10g, yeast extract 1～3g, dipotassium hydrogen phosphate 0.2～0.8g, magnesium sulfate 0.01～ 0.03g, calcium chloride 0.1～0.29g.
8. preparation method as claimed in claim 6, it is characterised in that in described step (1), (2) and step (3), two grades of kinds Sub-condition of culture is：Illumination 1000～3000LX, cultivation temperature 25～30 DEG C, Anaerobic culturel 3～4 days；
Every liter of component of described secondary seed medium is as follows：
Glucose 15～20g, peptone 6～10g, yeast extract 1～3g, dipotassium hydrogen phosphate 0.2～0.8g, magnesium sulfate 0.01～ 0.03g, calcium chloride 0.1～0.29g.
9. preparation method as claimed in claim 6, it is characterised in that in described step (1), (2) and step (3), expand training Foster condition is：Cultivation temperature 28～30 DEG C, dark aerobic cultivation 6～8 days, ratio of ventilating per hour is liquid and gas volume ratio 1:0.5, speed of agitator 200～300r/min；
The culture medium every liter component that described expansion is cultivated is as follows：
Glucose 15～20g, peptone 6～10g, yeast extract 1～3g, dipotassium hydrogen phosphate 0.2～0.8g, magnesium sulfate 0.01～ 0.03g, calcium chloride 0.1～0.29g.
10. process the application of the microbial bacterial agent of heavy metal pollution of soil described in claim 1, it is characterised in that step is as follows：
The microbial bacterial agent processing heavy metal pollution of soil is directly applied in the soil of heavy metal pollution as base manure, turns over Mixing, amount of application is 8～12g/ square metre, processes 33～38 days under conditions of temperature 29～31 DEG C, pH7.5～8.5.
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