CN110142285B - Method for remedying heavy metal contaminated soil by combination of trichoderma asperellum and alfalfa - Google Patents
Method for remedying heavy metal contaminated soil by combination of trichoderma asperellum and alfalfa Download PDFInfo
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Abstract
The invention discloses a method for remedying heavy metal contaminated soil by combining trichoderma asperellum and alfalfa; the contaminated soil is repaired by the synergistic adsorption of heavy metals in the soil by trichoderma asperellum and alfalfa. The trichoderma asperellum GDSF1009 capable of colonizing in alfalfa rhizosphere is prepared into granules and applied to alfalfa rhizosphere. Because the trichoderma granules can stimulate the alfalfa root system and improve the heavy metal adsorption capacity, the trichoderma granules and the alfalfa root system form a restoration biological complex for efficiently adsorbing heavy metals. The adsorption efficiency of heavy metal in the plant body reaches 51-59%, and the content of heavy metal in the soil is reduced by 58-60%. Meanwhile, the repaired plant material can be recycled through cleaning. The technology can effectively reduce the heavy metal content of the soil, can restore the soil micro-ecology, increases beneficial microorganisms in the soil and improves the soil fertility, and is a novel microorganism-plant combined soil remediation technology which is economical and effective and has wide application prospect.
Description
Technical Field
The invention belongs to the field of soil remediation, and particularly relates to a method for remediating heavy metal contaminated soil by combining trichoderma asperellum and alfalfa.
Background
The soil pollution is increasingly serious due to industrial pollution and excessive use of chemical fertilizers and pesticides, and the problem of severely restricting agricultural production is solved. In order to realize sustainable development of agriculture, the treatment of polluted soil is a major subject for restoring and improving ecological environment.
At present, the common heavy metal contaminated soil remediation technology mainly has three aspects: physical repair, chemical repair and microbial repair, phytoremediation, microbial-phytoremediation combined. The physical remediation method is a method for separating pollutants from soil by adopting certain technology and means to recover the available value of the soil. The following methods are mainly used for physical remediation of soil: direct soil replacement method, thermalization method repairing, vitrification repairing method, and electrode driving repairing method. The physical remediation cost is high, the operability is poor, and the large-area soil is difficult to be remedied. Chemical remediation is the addition of a passivating agent to contaminated soil to convert heavy metals from an active to a stabilized form to reduce heavy metal migration and bioavailability. However, chemical leaching easily causes soil acidification, breaks the physical structure of soil, causes loss of soil fertility, and easily causes secondary pollution to environments such as underground water and the like during chemical remediation. Bioremediation comprises the adsorption of heavy metals by enrichment plants and the adsorption of heavy metals by microorganisms, and the heavy metals are mainly solidified by generating chelating agents such as organic acid and the like. At present, most of the plants are singly utilized and are hyper-enriched internationally, such as vetiver, ciliate desert grass, sedge triquetrum, Indian mustard (Brassica juncea) and the like. Alfalfa is a perennial green manure leguminous plant with a nitrogen fixation function, so the alfalfa which is used as a repairable plant has the functions of adsorbing heavy metals and conserving soil. The plant enrichment method is economical and practical, and can improve the ecological environment, but the simple plant enrichment method cannot achieve high efficiency in restoring the heavy metal polluted soil. The microorganisms for repairing the soil are mainly used for enriching heavy metals through a metabolic system and subcells of the microorganisms, for example, trichoderma can adsorb heavy metal Cu through a purine system, trichoderma vacuoles can also enrich heavy metals (Fu Kehe and the like, 2014), and meanwhile, organic acids generated by the trichoderma can also solidify or chelate the heavy metals. Besides, the trichoderma has the repairing functions of degrading a series of organophosphorus pesticides and reducing soil salinization and the like. The research shows that: the adsorption efficiency of the mustard seedlings treated by the trichoderma atroviride F6 spore suspension on Cd and Ni is obviously improved (LixiangCao et al, 2008). Therefore, the combined use of the microorganism and the enrichment plant can generate a plurality of comprehensive effects of soil remediation.
Patent CN109174937A proposes a combined remediation system for remedying heavy metal contaminated soil by using a complex microbial inoculum to cooperate with organic fertilizers and chrysanthemum. However, this patent does not suggest a feasible method for treating plants enriched in heavy metals, nor does it suggest returning the plants grown as green manure to the field for soil conservation. The invention provides an effective method for subsequently treating heavy metal-enriched plants and returning the plants as green manure to the field, which can effectively reduce the heavy metal content of soil, restore the soil micro-ecology, increase beneficial microorganisms in the soil and improve the soil fertility, and is a novel microorganism-plant combined soil remediation technology with economy, effectiveness and wide application prospect.
Disclosure of Invention
The invention aims to overcome the defects in the prior art and provides a method for repairing heavy metal contaminated soil by combining trichoderma asperellum and alfalfa. The method is carried out by a Trichoderma asperellum GDSF1009 (journal of plant protection 2015,42(6):1030-1035) with the deposit number: the granules of CGMCC 9512) interact with the root system of alfalfa specifically, so that the efficiency of adsorbing heavy metal by alfalfa is improved, and meanwhile, the alfalfa straws adsorbing heavy metal are subjected to clean elution or solidification by organic acid and then are degraded by cellulase generated by trichoderma; preparing the straw bacterial manure.
The purpose of the invention is realized by the following technical scheme:
the invention relates to a method for repairing heavy metal contaminated soil, which adopts trichoderma asperellum and alfalfa to jointly repair the heavy metal contaminated soil.
In the invention, the trichoderma asperellum is trichoderma asperellum GDSF 1009; the alfalfa is alfalfa.
As an embodiment of the invention, the sowing amount of the alfalfa in the heavy metal polluted soil per mu is 1.0-3.0kg, and the application amount of the trichoderma asperellum granules is 5-20 kg. Under the compound dosage, the trichoderma asperellum granules are suitable for promoting the growth of (purple) alfalfa and improving the capacity of (purple) alfalfa to adsorb heavy metals in soil.
As an embodiment of the invention, the trichoderma asperellum granules are mixed with an organic fertilizer and then applied.
The application amount of the organic fertilizer is 50-150kg per mu.
In the invention, the trichoderma asperellum granules are prepared by the following method: mixing Trichoderma asperellum liquid fermentation broth 48-52 wt%, corn flour 13.625-17.625 wt%, diatomite 14.75-20.75 wt%, wheat bran 13.625-17.625 wt%, humic acid 0.4-0.6 wt%, citric acid 0.4-0.6 wt%, stirring, extruding, granulating, and drying.
Preferably, the drying is at 47 ℃ for 1.5 h.
The trichoderma asperellum liquid fermentation liquid is prepared by the following method: sterilizing the fermentation culture medium at 121 deg.C for 30-60min, inoculating 0.5-2.0 wt% of Trichoderma asperellum strain, controlling fermentation temperature at 25-35 deg.C, and fermenting for 5-10d until the fermentation liquid turns green.
The fermentation medium contains per 221.5709 kg: 5-15kg of corn flour, 220kg of water 200-.
Preferably, when the fermentation medium is prepared, the inorganic salts of potassium dihydrogen phosphate, magnesium sulfate, manganese sulfate, zinc sulfate, sodium nitrate, ammonium sulfate and sodium chloride are dissolved in water, and the corn flour and the water are fully mixed to prepare the fermentation medium.
The trichoderma asperellum subspecies is prepared by the following method: inoculating the trichoderma asperellum into a PDA culture medium to be cultured to obtain an activated trichoderma asperellum strain; inoculating activated trichoderma asperellum strains into a liquid fermentation strain culture medium, and fermenting to obtain the trichoderma strains.
Preferably, the liquid fermentation strain culture medium comprises the following components:
potato 200 g
Glucose 20g
1L of water
Adjusting pH to 6-8.
Preferably, the culturing is specifically as follows: inoculating Trichoderma asperellum GDFS1009 into PDA culture medium plate, culturing in 28 deg.C incubator for 2-3 d.
Preferably, the fermentation is specifically: and (3) filling 100ml of liquid fermentation strain culture medium into each bottle in a 250 ml triangular flask, sterilizing for 30min at 121 ℃, inoculating activated trichoderma asperellum strains, wherein 5 strain cakes are arranged in each bottle, the rotating speed of a shaking table is 180rpm, the fermentation temperature is 28 ℃, and the fermentation time is 3d, so as to obtain liquid fermentation seed bacteria, namely trichoderma asperellum strain seed bacteria.
As an embodiment of the invention, the method further comprises the steps of carrying out heavy metal elution treatment and neutralization treatment on the alfalfa straws repaired with the heavy metal contaminated soil in sequence, mixing the alfalfa straws with the trichoderma granules, stacking, covering with a film, and fermenting to prepare the straw bacterial manure.
Preferably, the alfalfa straws are small sections (preferably with the length of 0.5-2cm) cut from alfalfa stems, leaves and roots which absorb heavy metals.
Preferably, the heavy metal elution treatment is to treat the alfalfa straws for 30-50 hours by using citric acid solution with the concentration of 0.1-0.5 mol/L.
Preferably, the neutralization treatment is to soak the alfalfa straws after the heavy metals are eluted with clear water for 2-3 times, and each time lasts for 1-2 hours.
Preferably, the mixing ratio of the neutralized alfalfa straws to the trichoderma asperellum granules is 80-120: 1 w/w.
Preferably, the trichoderma asperellum granules have a cellulase activity (CMC) enzyme activity of 4.0U.g-1The above.
Preferably, the composting and film covering, and the fermenting specifically comprise: stacking 8-12cm straw layer on the ground surface, covering with film (with air holes), and fermenting at 25-27 deg.C for 20-30 d.
Compared with the prior art, the invention has the following beneficial effects:
1) compared with a single plant restoration system, the heavy metal restoration system has the advantages that the heavy metal adsorption capacity of plants is remarkably improved, beneficial microorganisms in soil are increased, and a soil micro-ecological system is restored;
2) compared with the existing microorganism-plant composite remediation system, the effective method for treating the heavy metal-enriched plants and returning the heavy metal-enriched plants to the field as green manure is provided, so that the heavy metal-enriched plants can be effectively treated, and the heavy metal-enriched plants can be returned to the field to improve the soil fertility.
Drawings
Other features, objects and advantages of the invention will become more apparent upon reading of the detailed description of non-limiting embodiments with reference to the following drawings:
FIG. 1 is a schematic diagram showing the average content of heavy metals in alfalfa and rape bodies after 5 months of planting in Pudong test fields.
Detailed Description
The present invention will be described in detail with reference to examples. The following examples will assist those skilled in the art in further understanding the invention, but are not intended to limit the invention in any way. It should be noted that it would be apparent to those skilled in the art that several modifications and improvements can be made without departing from the inventive concept. All falling within the scope of the present invention.
Example 1
A place: test site for Jinshandong Zhang Weizhen mulberry garden village electroplating plant
The embodiment relates to a method for repairing heavy metal contaminated soil by combining trichoderma asperellum and alfalfa.
Firstly, a preparation method of trichoderma granules is provided, which comprises the following steps:
1. plate strain: inoculating trichoderma asperellum GDFS1009 in a PDA culture medium plate, and in an incubator at 28 ℃; culturing in inverted culture for 2-3 days.
PDA culture medium: cutting 200 g potato into 1cm2The small blocks are boiled for 30 minutes by small fire, 4 layers of gauze are filtered, and juice is reserved; adding 20g of glucose, 20g of agar powder and distilled water to a constant volume of 1 liter, and sterilizing for 30 minutes by high-pressure steam at 121 ℃.
2. Preparing liquid strains: preparing a secondary fermentation culture solution (namely a liquid fermentation strain culture medium) by taking 200 g of potatoes, 20g of glucose and 1L of water, adjusting the pH value to 6-8, subpackaging the secondary fermentation culture solution in a 250 ml triangular flask with 100ml, sterilizing the secondary fermentation culture solution for 30 minutes at 121 ℃, inoculating a flat strain (PDA for culture, taking 5 fungus cakes in each bottle by using a 7mm puncher along the edges of the bacterial colony, rotating the shaking table at 180 r/min, fermenting the secondary fermentation culture solution at 28 ℃ for 3 days, and obtaining a large amount of dispersed flocculent hyphae and dispersed conidia liquid strains.
3. Preparing trichoderma liquid fermentation liquor: the fermentation medium comprises the following components: 10kg of corn flour, 210 kg of water, 766 g of monopotassium phosphate, 100 g of magnesium sulfate, 0.5g of manganese sulfate, 0.4 g of zinc sulfate, 284 g of sodium nitrate, 220g of ammonium sulfate and 200 g of sodium chloride, firstly dissolving inorganic salt (monopotassium phosphate, magnesium sulfate, manganese sulfate, zinc sulfate, sodium nitrate, ammonium sulfate and sodium chloride) in water, then fully mixing the culture material (corn flour) with the water to prepare a solid fermentation culture medium, filling the solid fermentation culture medium into a 300-liter fermentation tank, sterilizing at 121 ℃ for 30-60 minutes, inoculating 0.5% by weight of secondary strain, controlling the fermentation temperature to be 28 ℃, fermenting for 7 days until the culture material becomes green, namely trichoderma liquid fermentation broth;
4. preparation of trichoderma granules: the mass ratio is as follows: 50 wt% of trichoderma liquid fermentation liquor, 15.625 wt% of corn flour, 17.75 wt% of diatomite, 15.625 wt% of wheat bran, 0.5 wt% of humic acid and 0.5 wt% of citric acid, uniformly stirring by using a stirring machine, extruding and granulating by using a granulator, and drying for 1.5 hours at 47 ℃ by using a dryer to obtain the finished product, namely the trichoderma granule.
5. And (3) detecting the soil heavy metal pollution back bottom: every three-party organization detects according to the national standard method;
6. the treatment method of the soil remediation experimental group comprises the following steps:
alfalfa WL525HQ (Beijing Zhengdao ecology technology Co., Ltd.). 1.5kg of alfalfa seeds are sown in each mu of land, and 90kg of organic fertilizer (provided by Shanghai Guangming Holstein animal husbandry Co., Ltd.) and 10kg of trichoderma granules are applied at the same time. And (5) harvesting after the alfalfa is ripe.
7. The treatment method for repairing the soil control group comprises the following steps:
blank control group: sowing 1.5kg of alfalfa seeds per mu
Trichoderma powder control group: 1.5kg of alfalfa seeds are sown in each mu, and 5kg of trichoderma powder (provided by Shandong Tyno pharmaceutical Co., Ltd.) is applied at the same time;
organic fertilizer control group: 1.5kg of alfalfa seeds are sown in each mu, and 90kg of organic fertilizer (provided by Shanghai Guangsheng Stent animal husbandry Co., Ltd.) is applied at the same time;
8. detecting the soil remediation effect;
and (3) treatment of the soil sample: collecting soil samples with depth of 15-20cm from each treated plot by five-point sampling method, drying at 85 deg.C to constant weight, grinding, sieving with 100 mesh sieve, and bagging. Weighing a certain amount (about 0.025 g) of soil sample, digesting the sample by a hydrochloric acid-nitric acid-perchloric acid wet method, and analyzing and detecting the content of heavy metal in the soil by an inductively coupled plasma emission spectrometer (Perkin Elmer PE Optima 7000 in the United states). The data obtained were subjected to mathematical statistical analysis using SPSS20.0 software.
The calculation method comprises the following steps:
the soil heavy metal reduction rate (%) - (the heavy metal content in the soil before treatment-the heavy metal content in the soil after treatment)/the content of the heavy metal in the soil before treatment
The soil heavy metal reduction increase rate (%) (heavy metal content of control group-heavy metal content of treatment group)/heavy metal content of control group
After the soil is treated for 12 months, the heavy metal content of the soil is averagely reduced by more than 40 percent (Table 1).
TABLE 1 reduction rate of heavy metals in soil treated with Trichoderma inoculant and organic fertilizer
Note: and calculating according to the measured value in the soil.
Example 2
A place: pudong area and Qingcheng town factory electroplating plant test site
The embodiment relates to a method for repairing heavy metal contaminated soil by combining trichoderma asperellum and alfalfa.
Firstly, a preparation method of trichoderma biological granules is provided, which comprises the following steps:
1. plate strain: inoculating Trichoderma asperellum GDFS1009 into a PDA culture medium plate, performing inverted culture in an incubator at 28 ℃ for 2-3 days;
PDA culture medium: cutting 200 g of potatoes into small pieces of 1cm2, boiling for 30 minutes with soft fire, filtering with 4 layers of gauze, and keeping juice; adding 20g of glucose, 20g of agar powder and distilled water to a constant volume of 1 liter, and sterilizing for 30 minutes by high-pressure steam at 121 ℃;
2. preparing liquid strains: preparing a secondary fermentation culture solution by taking 200 g of potatoes, 20g of glucose and 1L of water, adjusting the pH value to 6-8, subpackaging 100ml of the secondary fermentation culture solution in a 250 ml triangular flask, sterilizing at 121 ℃ for 30 minutes, inoculating a flat strain, and obtaining a large amount of dispersed flocculent hyphae and dispersed conidium liquid strains when the rotating speed of a shaking table is 180 revolutions per minute, the fermentation temperature is 28 ℃ and the fermentation time is 3 days;
3. preparing trichoderma liquid fermentation liquor: the fermentation medium comprises the following components: 10kg of corn flour, 210 kg of water, 766 g of monopotassium phosphate, 100 g of magnesium sulfate, 0.5g of manganese sulfate, 0.4 g of zinc sulfate, 284 g of sodium nitrate, 220g of ammonium sulfate and 200 g of sodium chloride, firstly dissolving inorganic salt in the water, then fully mixing culture materials with the water to prepare a solid fermentation culture medium, filling the solid fermentation culture medium into a 300-liter fermentation tank, sterilizing at 121 ℃ for 30-60 minutes, inoculating 0.5% by weight of secondary strains, controlling the fermentation temperature to be 28 ℃, and fermenting for 7 days until the culture materials become green, namely trichoderma liquid fermentation broth;
4. preparation of trichoderma granules: the mass ratio is as follows: 50 wt% of trichoderma liquid fermentation liquor, 15.625 wt% of corn flour, 68.75 wt% of diatomite, 15.625 wt% of wheat bran, 0.5 wt% of humic acid and 0.5 wt% of citric acid, uniformly stirring by using a stirrer, extruding and granulating by using a granulator, and drying for 1.5 hours at 47 ℃ by using a dryer to obtain a finished product, namely the trichoderma granule.
5. The treatment method of the soil remediation experimental group comprises the following steps:
alfalfa WL525HQ (Beijing Zhengdao ecology technology Co., Ltd.). 1.5kg of alfalfa seeds and 10kg of trichoderma granules are sown in each mu of land. And (5) harvesting after the alfalfa is ripe.
6. The treatment method for repairing the soil control group comprises the following steps:
and (3) rape CK group: rape (Huyou No. 6). Sowing 0.6kg of rape seeds in each mu of land, and harvesting after rape is mature;
rape + trichoderma granule group: rape (Huyou No. 6). 0.6kg of rape seeds and 10kg of trichoderma granules are sown in each mu of land. Harvesting after rape is ripe
Alfalfa CK group: alfalfa WL525HQ (Beijing Zhengdao ecology technology Co., Ltd.). 1.5kg of alfalfa seeds are sown in each mu of land. And (5) harvesting after the alfalfa is ripe. (Experimental Steps 5-6 all treatments were conducted with a single amount of organic fertilizer)
7. The method for detecting the content of heavy metals in the bodies of the alfalfa and the rape is characterized in that the alfalfa and the rape are harvested after 5 months of planting, and the processing method comprises the following steps:
collecting plant samples of different treatments, washing with distilled water, removing water on the surface with absorbent paper, dividing into root, stem and leaf, deactivating enzyme in a 120 deg.C oven, and oven drying at 85 deg.C for 24 hr. And crushing the dried plant sample by a crusher, sieving the crushed plant sample by a 100-mesh sieve, weighing about 0.03g, and bagging for later use. The sample is digested by a hydrochloric acid-nitric acid-perchloric acid wet method, and the heavy metal content in the plant sample is analyzed and detected by an inductively coupled plasma emission spectrometer (American Perkin Elmer PE Optima 7000). The data obtained were subjected to mathematical statistical analysis using SPSS20.0 software.
See fig. 1. The accumulation amount of heavy metals of Cu and Zn in alfalfa bodies is higher than that of 120.15% and 256.82% of the control groups. The accumulation amount of the Cu and Zn heavy metals in the rape bodies is 71.51 percent and 190.43 percent higher than those in the control group.
Example 3
The embodiment relates to a method for preparing straw bacterial manure after repairing heavy metal contaminated soil by combining trichoderma asperellum and alfalfa. The method comprises the following steps:
1. harvesting alfalfa straws: the stems, leaves and roots of the alfalfa which can absorb heavy metals (the alfalfa straws are cut into small sections with the length of 1cm by a pulverizer);
2. and (3) eluting heavy metals: treating root and stem tissue of alfalfa with 0.1-0.5 mol/L citric acid solution with different concentrations for 40h to obtain heavy metal elution rate of over 40% (Table 2). In addition, 100kg of 0.5mol of industrial citric acid is put into a straw cleaning treatment pool (side length multiplied by depth: 2m multiplied by 1m) and can be repeatedly used.
TABLE 2 elution effect of alfalfa root and stem heavy metal zinc after citric acid immersion
3. Performing alfalfa straw neutralization treatment: the alfalfa straws which are eluted with the organic acid solution to remove heavy metals are soaked in clear water for 2-3 times, each time lasts for 1-2 hours, and the enrichment level of the heavy metals in alfalfa tissues is obviously lower than that before the heavy metals are not treated by more than 80 percent, and the pH value is 6-7.
4. Preparation of cellulase Activity (CMC) with an enzyme Activity of 4.0U.g-1The above Trichoderma granules;
5. degrading alfalfa straws: mixing the cleaned alfalfa straw sections with trichoderma granules (100: 1w/w), stacking a 10cm straw layer on the ground surface, covering a film (with air vents), and fermenting at 25-27 ℃ for 20-30 days to prepare the straw bacterial fertilizer.
In conclusion, the contaminated soil is repaired by the trichoderma asperellum and the alfalfa to synergistically adsorb the heavy metals in the soil. The trichoderma asperellum GDSF1009 capable of colonizing in alfalfa rhizosphere is prepared into granules and applied to alfalfa rhizosphere. Because the trichoderma asperellum granules can stimulate the alfalfa root system and improve the heavy metal adsorption capacity of the alfalfa root system, the trichoderma asperellum granules and the alfalfa root system form a remediation biological complex for efficiently adsorbing heavy metals. The adsorption efficiency of heavy metal in the plant body reaches 51-59%, and the content of heavy metal in the soil is reduced by 58-60%. Meanwhile, the repaired plant material can be recycled through cleaning. The technology can effectively reduce the heavy metal content of the soil, can restore the soil micro-ecology, increases beneficial microorganisms in the soil and improves the soil fertility, and is a novel microorganism-plant combined soil remediation technology which is economical and effective and has wide application prospect.
The foregoing description of specific embodiments of the present invention has been presented. It is to be understood that the present invention is not limited to the specific embodiments described above, and that various changes and modifications may be made by one skilled in the art within the scope of the appended claims without departing from the spirit of the invention.
Claims (8)
1. A method for restoring heavy metal contaminated soil is characterized in that trichoderma asperellum and alfalfa are adopted to jointly restore the heavy metal contaminated soil; the trichoderma asperellum is trichoderma asperellum GDSF 1009; the alfalfa is alfalfa; the heavy metal contaminated soil contains one or more of mercury, lead, zinc and copper;
the applied trichoderma asperellum granules are prepared by the following method: mixing Trichoderma asperellum liquid fermentation broth 48-52 wt%, corn flour 13.625-17.625 wt%, diatomite 14.75-20.75 wt%, wheat bran 13.625-17.625 wt%, humic acid 0.4-0.6 wt%, citric acid 0.4-0.6 wt%, stirring, extruding, granulating, and drying.
2. The method for remediating heavy metal contaminated soil as recited in claim 1, wherein the seeding amount of the alfalfa is 1.0-3.0kg per mu of the heavy metal contaminated soil, and the application amount of the trichoderma asperellum granules is 5-20kg per mu of the heavy metal contaminated soil.
3. The method for remediating heavy metal contaminated soil as recited in claim 2, wherein the trichoderma asperellum granules are mixed with an organic fertilizer and applied.
4. The method for remediating heavy metal contaminated soil as recited in claim 3, wherein the application amount of the organic fertilizer is 50-150kg per mu.
5. The method for remediating heavy metal contaminated soil according to claim 1, wherein the trichoderma asperellum liquid fermentation broth is prepared by a method comprising: sterilizing the fermentation culture medium at 121 deg.C for 30-60min, inoculating 0.5-2.0 wt% of Trichoderma asperellum strain, controlling fermentation temperature at 25-35 deg.C, and fermenting for 5-10d until the fermentation liquid turns green.
6. The method for remediating heavy metal contaminated soil as recited in claim 5, wherein said fermentation medium contains, per 221.5709 kg: 5-15kg of corn flour, 220kg of water 200-.
7. The method for remediating heavy metal contaminated soil as recited in claim 5, wherein said Trichoderma asperellum subspecies is prepared by the following method: inoculating the trichoderma asperellum into a PDA culture medium to be cultured to obtain an activated trichoderma asperellum strain; inoculating activated trichoderma asperellum strains into a liquid fermentation strain culture medium, and fermenting to obtain the trichoderma strains.
8. The method for remediating heavy metal contaminated soil as recited in claim 1, further comprising the steps of sequentially subjecting alfalfa straw subjected to heavy metal elution and neutralization treatment, mixing with trichoderma granules, stacking, laminating, and fermenting to obtain straw bacterial manure; the heavy metal elution treatment is to treat the alfalfa straws for 30-50 hours by using citric acid solution with the concentration of 0.1-0.5 mol/L; the neutralization treatment is to soak the alfalfa straws after the heavy metals are eluted with clear water for 2-3 times, and each time lasts for 1-2 hours.
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