CN104059855A - Composite fungicide for treating heavy metal pollution of soil and preparation method of composite fungicide - Google Patents
Composite fungicide for treating heavy metal pollution of soil and preparation method of composite fungicide Download PDFInfo
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Abstract
The invention discloses a composite fungicide containing following four bacterial strains: Epicoccum nigrum, Leptosphaerulina chartarum, Lecanicillium sp. and Peyronellaca pomorum. The fungicide can be used for treating heavy metal pollution of soil. According to the invention, the fungicide is prepared from mycothallus instead of bacteria, and the unit biomass of the fungicide is far higher than that of a bactericide, so that the fungicide is better in adsorption effect; in addition, the fungicide can be cultured in a solid state, and a finished product of the fungicide can be solid particles, so that the fungicide is more convenient to use.
Description
Technical field
The present invention relates to microorganism field, relate in particular to a kind of composite fungi preparation of administering heavy metal pollution of soil and preparation method thereof.
Background technology
Along with the rapid expansion of mankind's process of industrialization, the heavy metal pollution problem of soil is the large difficult problem urgently to be resolved hurrily of the current field of environment protection one of China.Heavy metal pollution of soil problem is essential, involves the normal use in water source, soil, and food safety is the significant problem that is related to Environmental Health, people life property safety.The large-area pollution in the whole nation, the pollution new region, the pollution problem complicated and changeable that occur fast make traditional improvement technological method cannot reach satisfied regulation effect.Therefore, we need a kind of modern technique of more efficient, with low cost, environmentally friendly harmless, low maintenance.Along with people for microbial world understanding progressively deeply, scientist starts microorganism for administering environment, this be environmental protection more, low-carbon (LC), ecological friendly, merge natural method.
Traditional methods for curing heavy metal contamination in soil belongs to physics, chemical category substantially.Physical method comprises soil replacement method, soil moved in to improve the original method, electrodynamics method, thermal desorption method, extraction method etc.Soil replacement method is to change new free of contamination soil after the topsoil of pollution is shifted; Soil moved in to improve the original method is in contaminated soil, to add a large amount of pollution-free soil, covers top layer or mixes.Electrodynamics method is at soil applying direct current electric field, by effects such as electrolysis, electromigration, diffusion, electric osmose, electrophoresis, heavy metal ion is done relative motion, and at one end electrode is collected heavy metal.Thereby thermal desorption method carries out heat temperature raising to soil leaves edatope by heavy metal by volatilization.Extraction method is utilized the heavy metal effect in reagent and soil, forms soluble substance, then water rushes pollutent to root topsoil, adds certain medicament and heavy metal to form to precipitate and finally extracts.Chemical process comprises and applies modifying agent, adds reductive organic matter etc.Modifying agent refers generally to alkaline matter (as calcareous material), adds modifying agent that soil pH value is raise, and reduces the solvability of heavy metal, thereby can reduce plant for the absorption of heavy metal in soil.Add reductive agent can make some heavy metal in soil change valence state, maintain the form that toxicity is lower.Wherein, the characteristic repairing heavy metal pollution that utilizes microorganism is current study hotspot, but most of research is all in initial stage, does not produce the ripe actual technological method that can be applicable to.Recklessly former (2009, application publication number: CN101838086) disclose a kind of biotechnological formulation and by the method for biotechnological formulation lignin-sludge.This technology has adopted the mixed preparation of Sucus Cocois and calcium carbonate to process biological activity mud.Sun Hongwen etc. (2009) disclose the biological compound formulation that a kind of repairing heavy metal in soil pollutes.This technology has been utilized the characteristic of the Adsorption of Heavy Metals of a kind of bacteria culture-subtilis, and this bacterial suspension and a certain amount of humic acids are added in fertilizer, has played fixedly heavy metal and the effect that improves farmland.
In the prior art, there is the problem that quantities is large and secondary pollution is serious in soil removal and replacement soil moved in to improve the original method, therefore only limits to the IA that area is less; Electronic force method current consumption is huge, and cost is too expensive; Thermal desorption method need to provide a large amount of heat energy, so energy consumption is large, has also greatly improved cost.Chemical method need to add a large amount of chemical agents mostly, and not only cost is expensive, and very easily produces a large amount of mud, causes secondary pollution.The growth cycle that phytoremediation in biological restoration needs is very long, takes effect slow, simultaneously also very difficult for choosing of the super enriching plant of different heavy metals.And the mature technology that utilizes microbiological treatment heavy metal pollution of soil seldom, patent is also less.Former patented technology has been utilized the inner microflora self existing of mud recklessly, does not cultivate and produces specific microbe colony, therefore can only be applied to containing in the mud of microbe composition, and cannot be applicable to other soil types, and range of application is narrow.The patent of Sun Hongwen has been utilized bacillus subtilis bacteria suspension, and Dan Qi unit's biomass and adsorption effect are unsatisfactory.
Therefore, in this area, need unit biomass high, advantages of good adsorption effect, the microbial preparation of improvement heavy metal pollution of soil easy to use.
Summary of the invention
The limitation and the deficiency that the object of the invention is to overcome the traditional method of existing improvement heavy metal pollution of soil, provide a kind of environmental protection, non-secondary pollution, with low cost, the easy to use composite fungi preparation that utilizes to administer the technology of heavy metal pollution of soil and the preparation method of composite fungi preparation.
For reaching this object, the present invention by the following technical solutions:
In first aspect, the invention provides a kind of composite fungi microbial inoculum, it is characterized in that, described microbial inoculum contains Epicoccum nigrum, Leptosphaerulina chartarum, Lecanicillium sp. and tetra-kinds of bacterial classifications of Peyronellaca pomorum.
Described microbial inoculum of the present invention is solid.Preferably in described microbial inoculum, the particle diameter of thalli granule can be 1-3mm, more preferably 2mm.
In microbial inoculum of the present invention, the mass ratio of described four kinds of bacterial classifications can be 1:1:1:1:1.
In second aspect, the invention provides the production method of microbial inoculum as described in first aspect, it is characterized in that, described method comprises cultivates respectively described four kinds of bacterial classifications, afterwards four kinds of cultivated bacterial classifications is mixed.
In the production method of described microbial inoculum of the present invention, described method of cultivating respectively described four kinds of bacterial classifications can comprise:
(a) first order seed is cultivated;
(b) secondary seed is cultivated;
(c) enlarged culturing;
(d) collect thalline;
(e) optionally, thalline is dry; With
(f) optionally, thalline is pulverized.
In the production method of described microbial inoculum of the present invention, described enlarged culturing can be solid enlarged culturing, preferably described enlarged culturing can be carried out take glucose and peptone in main Solid agar culture, and preferably, described enlarged culturing can be carried out in the circular culture dish of large glass.
The production method of described microbial inoculum of the present invention can comprise:
(1) described first order seed is cultivated as the cultivation of slant tube first order seed:
Prepare four groups of aseptic slant tube substratum, its formula 1kg substratum comprises: be glucose 18～23g, peptone 8～12g, agar 12～16g, sodium-chlor 0.1～0.3g, calcium chloride 0.1～0.2g, Repone K 0.3～0.6g, surplus is water, described four kinds of bacterial classifications of separation and purification are seeded to respectively respectively to organize and in inclined-plane, carry out single culture, 23 ℃-27 ℃ of culture temperature, preferably 24 ℃-26 ℃, most preferably 25 ℃, cultivate 4-6 days, 4.5-5.5 days preferably, more preferably 5 days, thalline covers with inclined-plane, obtain respectively the first order seed of described four kinds of bacterial classifications,
(2) described secondary seed is cultivated as the cultivation of Petri dish secondary seed:
Prepare four groups of aseptic Petri Dish substratum, its formula is for containing anhydrous sodium acetate 0.24-0.26g in 1L substratum, 0.245-0.255g preferably, 0.25g more preferably, yeast extract 0.12-0.16g, preferably 0.13-0.15g, 0.14g more preferably, agar 12-16g, preferably 13-15g, more preferably 14g, tetrahydrate manganese chloride 300-500 μ mol/L, 350-450 μ mol/L preferably, 400 μ mol/L more preferably, HEPES4.0-4.2g, 4.05-4.15g preferably, 4.1g more preferably, surplus is deionized water, pH is 5-7;
Under aseptic condition, in four groups of substratum, access respectively the first order seed of described four kinds of bacterial classifications, described four kinds of bacterial classifications are cultivated separately under separated condition, 23 ℃-27 ℃ of culture temperature, preferably 24 ℃-26 ℃, most preferably 25 ℃, incubation time is 7～10 days, preferably 8-9 days, more preferably 8.5 days, obtain the secondary seed of described four kinds of bacterial classifications;
(3) described enlarged culturing is large glass culture dish enlarged culturing:
Preparation substratum, its formula comprises for 1kg substratum: be glucose 18～23g, peptone 8～12g, agar 12～16g, sodium bicarbonate 0.15g, three water dipotassium hydrogen phosphate 0.35g, magnesium sulfate 0.05g, iron vitriol 0.1g, sodium-chlor 0.1～0.3g, calcium chloride 0.1～0.2g, Repone K 0.3～0.6g, surplus is water, makes sterile solid substratum, and described solid medium is divided and is filled in the circular culture dish of large glass;
Inoculation spreads cultivation: the secondary seed that accesses respectively described four kinds of bacterial classifications under aseptic condition in four groups of substratum, described secondary seed is taken in the circular culture dish of fritter access large glass, and described four kinds of bacterial classifications are cultivated separately under separated condition, and culture temperature is 23 ℃-27 ℃, preferably 24 ℃-26 ℃, most preferably 25 ℃, incubation time is 9～14 days, preferably 10-13 days, 11-12 days more preferably, pH is 5-8,6-7 preferably, more preferably 6.5;
(4) collect thalline:
Under aseptic condition, the thalline of described four kinds of bacterial classifications of having grown spreading cultivation is transferred to respectively independent aseptic clothes hanger;
(5) thalline is dry
Under aseptic condition, the natural air drying by the thalline on clothes hanger by loft drier is dried, 20 ℃-25 ℃ of drying temperatures, and preferably 21 ℃-24 ℃, more preferably 22 ℃-23 ℃, humidity is below 20%;
(6) thalline is pulverized
Under aseptic condition, the thalline of described four kinds of bacterial classifications of natural air drying is put into respectively to sterile crushing machine and carry out crushing operation, obtain the pulverizing thalline of described four kinds of bacterial classifications;
(7) mix thalline
Get respectively the pulverizing thalline of described four kinds of bacterial classifications, fully mix, obtain described microbial inoculum.
In the third aspect, the invention provides the method that the microbial inoculum described in first aspect is administered heavy-metal contaminated soil, described method comprises, the microbial inoculum the invention provides described in first aspect is contacted with described heavy-metal contaminated soil, and preferably described heavy metal contamination is manganese metallic pollution.
In the method for improvement heavy-metal contaminated soil of the present invention, the consumption of described microbial inoculum can be for more than 5g microbial inoculum/kg soil sample.
Beneficial effect of the present invention:
(1) the present invention has utilized mycothallus rather than bacterium, because the unit biomass of fungi will be far longer than bacterium, therefore the adsorption effect of composite fungus agent of the present invention is better
(2) the present invention can adopt solid-state cultivation, and the finished product of manufacturing can be solid particulate, uses convenient.
(3) in fungi preparation of the present invention, comprise four kinds of fungies, it cooperatively interacts and has the action effect of addition.
Accompanying drawing explanation
Fig. 1 is the adsorption efficiency variation diagram of the different microbial inoculum dosage of the present invention hypothallus to cadmium.
Fig. 2 is the adsorption efficiency variation diagram of the different microbial inoculum dosage of the present invention hypothallus to zinc.
Embodiment
Below by embodiment, further illustrate technical scheme of the present invention.
The production method of embodiment 1. composite fungi preparations
(1) slant tube first order seed is cultivated:
According to 1kg substratum, comprise glucose 18g, peptone 8g, agar 12g, sodium-chlor 0.1g, calcium chloride 0.1g, Repone K 0.3g, the formulated slant tube substratum that surplus is water.Make four groups of slant tube substratum, four kinds of bacterial classifications of separation and purification are seeded to respectively respectively to organize and in inclined-plane, carry out single culture.Culture temperature is controlled at 23-25 ℃, and after 4-6 days, thalline covers with inclined-plane.Test tube after slant culture finishes is placed on preservation in 4 ℃ of refrigerators.
(2) Petri dish secondary seed is cultivated:
Substratum preparation: in preparation 1L substratum, contain anhydrous sodium acetate 0.24-0.26g, yeast extract 0.12-0.16g, agar 12-16g, tetrahydrate manganese chloride 300-500 μ mol/L, HEPES4.0-4.2g, deionized water 1L, being adjusted to pH is 7.By substratum sterilizing 30-40min under the condition of 120 ℃ of temperature.The culture medium solution taking out from autoclave sterilization pot need to be cooled to 60 ℃ in 60 ℃ of water-baths of constant temperature.The 20mL that cooled substratum is got equivalent divides in the circular culture dish of disposable Corning that is filled to the high 20mm specification of diameter 100mm, after cooled and solidified, with sealed membrane sealing, under 4 ℃ of conditions, preserves.
The cultivation of secondary seed: the Petri Dish substratum equivalent of producing is divided into four groups, accesses respectively four kinds of bacterial classifications under aseptic condition in four groups of substratum, four kinds of bacterial classifications are cultivated separately under separated condition.24～27 ℃ of culture temperature, incubation time are 7～10 days, and substratum bacterium state smooth surface is without microbiological contamination.
(3) large glass culture dish enlarged culturing:
Substratum preparation: glucose 18g, peptone 8g, agar 12g, sodium bicarbonate 0.15g, three water dipotassium hydrogen phosphate 0.35g, magnesium sulfate 0.05g, iron vitriol 0.1g, sodium-chlor 0.1g, calcium chloride 0.1g, Repone K 0.3g, makes solid medium, every bottled amount is about 1Kg, sterilization 30min at 130 ℃.The culture medium solution taking out from autoclave sterilization pot need to be cooled to 60 ℃ in 60 ℃ of water-baths of constant temperature.The 70mL that cooled substratum is got equivalent divides in the circular culture dish of large glass that is filled to the high 30mm specification of diameter 200mm, after cooled and solidified, with sealed membrane sealing, under 4 ℃ of conditions, preserves.
Inoculation spreads cultivation: the circular substratum equivalent of the large glass of producing is divided into four groups, under aseptic condition, in four groups of substratum, access respectively four kinds of bacterial classifications, first order seed bacterial classification is taken in the circular culture dish of fritter access large glass, and four kinds of bacterial classifications are cultivated separately under separated condition.Culture temperature is 25～27 ℃, and incubation time is 9～14 days, and pH is about 5～8, after testing without entering next step processing after miscellaneous bacteria.
(4) thalline biomass is collected:
In environment after strict sterilization (after ozonization 2h again through uv sterilisation 30min), by spreading cultivation in large glass culture dish, the thalline of having grown is transferred to aseptic plastic clothes hanger, the collection process of every kind of fungi strain is consistent, but independent respectively clothes hanger need to be set, in process, stop crossed contamination, it is aseptic clean that whole process keeps.
(5) thalline is dry
In sterilisable chamber, by being placed on thalline on the clothes hanger natural air drying by loft drier, make it to reach drying regime.Four kinds of bacterial classifications are respectively equipped with independently loft drier.It is aseptic clean that whole drying process keeps, and room ventilated, through the filtration of air filter, is guaranteed sterile air inclusion-free particle.Temperature is controlled at 20-25 ℃, without extra heating system.Humidity is controlled at below 20%.
(6) thalline is pulverized
The thalline of four kinds of bacterial classifications of natural air drying is entered and in sterile crushing machine, carries out crushing operation.Grain diameter after pulverizing is in 1-3mm left and right.
(7) canned and storage
Get respectively four kinds of bacterial classifications of equivalent pulverizing thalline (the mass ratio of four kinds of bacterial classifications is 1:1:1:1, respectively account for total amount 25%), it is fully mixed, use aseptic wide-mouth brown glass reagent bottle to carry out the canned of thalli granule.After bottling, be placed under lucifuge, dry, room temperature condition and preserve.
Embodiment 2. adopts composite fungi preparation of the present invention to process the pedotheque containing high concentration manganese
Composite fungi preparation of the present invention is applied to Chongqing containing in the pedotheque of high concentration manganese, obtain good regulation effect (table 1).The consumption of composite fungi preparation is 5g preparation/kg soil sample.The soil of pollution by manganese is taken from the on-site topsoil of Chongqing manganese ore, gets altogether the soil in place everywhere as sample, everywhere in soil containing the concentration of manganese difference to some extent.As can be seen from Table 1, composite fungi preparation of the present invention is wider for tolerance range and the subject range of Manganese In Soils concentration, be no matter～10mg/kg or～100mg/kg containing manganese concentration, can both play good effect, clearance has all reached more than 96%.
Table 1: composite fungi preparation of the present invention is for the regulation effect containing wad earth
Embodiment 3. adopts composite fungi preparation of the present invention to process the pedotheque containing high density cadmium zinc
Taking and being subject to each 5g of cadmium pollution soil sample is blank, prepare the sample that 4 groups of composite fungi preparations are processed simultaneously, every group of sample is containing soil sample weight 5g, wherein add respectively microbial bacteria liquid measure than being 20ml, 30ml, 50ml, 100ml, the pH value of all samples is adjusted to 6.0, the adjusting of soil pH value adopts 1mol/L HCl or 1mol/L NaOH, incubated at room temperature.
Experimental result shows, the composite fungus agent being mixed by these four kinds of fungi strains has very strong fixed action for the cadmium in soil and zinc.After adding the microbial inoculum of different amounts, the cadmium zinc concentration in the soil solution has been compared decline significantly (table 2) with blank.Along with the increase of microbial inoculum dosage, thalline is to the adsorption efficiency of cadmium zinc decrease (Fig. 1,2).Particularly, while adding the microbial inoculum of 20-50mL scope, thalline is higher to the adsorption efficiency of cadmium, remains on 95% left and right.
Table 1: remaining cadmium zinc content in the soil solution under different microbial inoculum dosages
Applicant's statement, the present invention illustrates detailed features of the present invention and method by above-described embodiment, but the present invention is not limited to above-mentioned detailed features and method, does not mean that the present invention must rely on above-mentioned detailed features and method could be implemented.Person of ordinary skill in the field should understand; any improvement in the present invention; to the selection of the increase of the equivalence replacement of the selected material of the present invention and step and subsidiary material and step, concrete mode etc., within all dropping on protection scope of the present invention and open scope.
Claims (9)
1. a composite fungi microbial inoculum, it is characterized in that, described microbial inoculum contains epicoccum nigrum (Epicoccum nigrum, CGMCC No.5775), Leptosphaerulina chartarum (CGMCC No.5776), Lecanicillium sp. (CGMCC No.6337) and four kinds of bacterial classifications of Peyronellaca pomorum (CGMCC No.6338).
2. microbial inoculum according to claim 1, is characterized in that, described microbial inoculum is solid; Preferably, in described microbial inoculum, the particle diameter of thalli granule is 1-3mm, more preferably 2mm.
3. microbial inoculum according to claim 1 and 2, is characterized in that, the mass ratio of described four kinds of bacterial classifications is 1:1:1:1～3:3:2:1.
4. the production method of the microbial inoculum described in any one in claim 1-3, is characterized in that, described method comprises cultivates respectively described four kinds of bacterial classifications, afterwards four kinds of cultivated bacterial classifications is mixed.
5. method according to claim 4, is characterized in that, described method of cultivating respectively described four kinds of bacterial classifications comprises:
(a) first order seed is cultivated;
(b) secondary seed is cultivated;
(c) enlarged culturing;
(d) collect thalline;
(e) optionally, thalline is dry; With
(f) optionally, thalline is pulverized.
6. method according to claim 5, it is characterized in that, described enlarged culturing is solid enlarged culturing, and preferably described enlarged culturing is carried out take glucose and peptone in main Solid agar culture, and preferably, described enlarged culturing is carried out in the circular culture dish of large glass.
7. method according to claim 5, is characterized in that,
(1) described first order seed is cultivated as the cultivation of slant tube first order seed:
Prepare four groups of aseptic slant tube substratum, its formula 1kg substratum comprises: be glucose 18～23g, peptone 8～12g, agar 12～16g, sodium-chlor 0.1～0.3g, calcium chloride 0.1～0.2g, Repone K 0.3～0.6g, surplus is water, described four kinds of bacterial classifications of separation and purification are seeded to respectively respectively to organize and in inclined-plane, carry out single culture, 23 ℃-27 ℃ of culture temperature, preferably 24 ℃-26 ℃, most preferably 25 ℃, cultivate 4-6 days, 4.5-5.5 days preferably, more preferably 5 days, thalline covers with inclined-plane, obtain respectively the first order seed of described four kinds of bacterial classifications,
(2) described secondary seed is cultivated as the cultivation of Petri dish secondary seed:
Prepare four groups of aseptic Petri Dish substratum, its formula is for containing anhydrous sodium acetate 0.24-0.26g in 1L substratum, 0.245-0.255g preferably, 0.25g more preferably, yeast extract 0.12-0.16g, preferably 0.13-0.15g, 0.14g more preferably, agar 12-16g, preferably 13-15g, more preferably 14g, tetrahydrate manganese chloride 300-500 μ mol/L, 350-450 μ mol/L preferably, 400 μ mol/L more preferably, HEPES4.0-4.2g, 4.05-4.15g preferably, 4.1g more preferably, surplus is deionized water, pH is 5-7;
Under aseptic condition, in four groups of substratum, access respectively the first order seed of described four kinds of bacterial classifications, described four kinds of bacterial classifications are cultivated separately under separated condition, 23 ℃-27 ℃ of culture temperature, preferably 24 ℃-26 ℃, most preferably 25 ℃, incubation time is 7～10 days, preferably 8-9 days, more preferably 8.5 days, obtain the secondary seed of described four kinds of bacterial classifications;
(3) described enlarged culturing is large glass culture dish enlarged culturing:
Preparation substratum, its formula comprises for 1kg substratum: be glucose 18～23g, peptone 8～12g, agar 12～16g, sodium bicarbonate 0.15g, three water dipotassium hydrogen phosphate 0.35g, magnesium sulfate 0.05g, iron vitriol 0.1g, sodium-chlor 0.1～0.3g, calcium chloride 0.1～0.2g, Repone K 0.3～0.6g, surplus is water, makes sterile solid substratum, and described solid medium is divided and is filled in the circular culture dish of large glass;
Inoculation spreads cultivation: the secondary seed that accesses respectively described four kinds of bacterial classifications under aseptic condition in four groups of substratum, described secondary seed is taken in the circular culture dish of fritter access large glass, and described four kinds of bacterial classifications are cultivated separately under separated condition, and culture temperature is 23 ℃-27 ℃, preferably 24 ℃-26 ℃, most preferably 25 ℃, incubation time is 9～14 days, preferably 10-13 days, 11-12 days more preferably, pH is 5-8,6-7 preferably, more preferably 6.5;
(4) collect thalline:
Under aseptic condition, the thalline of described four kinds of bacterial classifications of having grown spreading cultivation is transferred to respectively independent aseptic clothes hanger;
(5) thalline is dry
Under aseptic condition, the natural air drying by the thalline on clothes hanger by loft drier is dried, 20 ℃-25 ℃ of drying temperatures, and preferably 21 ℃-24 ℃, more preferably 22 ℃-23 ℃, humidity is below 20%;
(6) thalline is pulverized
Under aseptic condition, the thalline of described four kinds of bacterial classifications of natural air drying is put into respectively to sterile crushing machine and carry out crushing operation, obtain the pulverizing thalline of described four kinds of bacterial classifications;
(7) mix thalline
Get respectively the pulverizing thalline of described four kinds of bacterial classifications, fully mix, obtain described microbial inoculum.
8. adopt the method that in claim 1-3, the microbial inoculum described in any one is administered heavy-metal contaminated soil, described method comprises, microbial inoculum described in any one in claim 1-3 is contacted with described heavy-metal contaminated soil, and preferably described heavy-metal contaminated soil is manganese metallic pollution soil.
9. method according to claim 8, the consumption of wherein said microbial inoculum is more than 5g microbial inoculum/kg soil sample.
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| CN105296359A (en) * | 2015-08-21 | 2016-02-03 | 浙江中烟工业有限责任公司 | Endophytic fungus strain NYN771C06 and application thereof |
| CN106591156A (en) * | 2017-01-16 | 2017-04-26 | 昆明理工大学 | Epicoccum nigrum FXZ2 and application thereof |
| CN106591156B (en) * | 2017-01-16 | 2019-07-16 | 昆明理工大学 | One plant of epicoccum nigrum FXZ2 and its application |
| CN108405603A (en) * | 2018-02-07 | 2018-08-17 | 深圳市深港产学研环保工程技术股份有限公司 | A method of leaching being carried out to heavy-metal contaminated soil using microorganism formulation |
| CN108405603B (en) * | 2018-02-07 | 2020-09-29 | 深圳市深港产学研环保工程技术股份有限公司 | Method for leaching heavy metal contaminated soil by using microbial preparation |
| CN110221041A (en) * | 2019-06-06 | 2019-09-10 | 生态环境部南京环境科学研究所 | The method for carrying out heavy-metal contaminated soil ecotoxicity assay using luminous earthworm |
| CN110221041B (en) * | 2019-06-06 | 2022-01-21 | 生态环境部南京环境科学研究所 | Method for detecting heavy metal contaminated soil ecotoxicity by using luminous earthworms |
| CN110577909A (en) * | 2019-09-17 | 2019-12-17 | 合肥师范学院 | method for preparing efficient phosphate solubilizing epicoccum with heavy metal tolerance characteristic |
| TWI761751B (en) * | 2020-01-14 | 2022-04-21 | 明志科技大學 | Method and equipment for removing contaminants by using mixed bacterial liquid to enhance electric power |
| CN112358974A (en) * | 2020-12-09 | 2021-02-12 | 昆明理工大学 | Endophytic fungus epicoccum nigrum FZT214 and application thereof |
| CN112358974B (en) * | 2020-12-09 | 2022-06-10 | 昆明理工大学 | Plant endophytic fungus epicoccum nigrum FZT214 and application thereof |
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