CN108160685B - Method for repairing coal mining area soil by combining plants and microorganisms - Google Patents
Method for repairing coal mining area soil by combining plants and microorganisms Download PDFInfo
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- CN108160685B CN108160685B CN201810000526.8A CN201810000526A CN108160685B CN 108160685 B CN108160685 B CN 108160685B CN 201810000526 A CN201810000526 A CN 201810000526A CN 108160685 B CN108160685 B CN 108160685B
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Classifications
-
- 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
-
- 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
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B09—DISPOSAL OF SOLID WASTE; RECLAMATION OF CONTAMINATED SOIL
- B09C—RECLAMATION OF CONTAMINATED SOIL
- B09C2101/00—In situ
Abstract
The invention belongs to the technical field of soil bioremediation, and particularly relates to a method for restoring coal mining area soil. A method for restoring the soil in coal mining area by plant-microbe combination includes such steps as insolating the seeds of Trifolium ramosissimum, immersing seeds, germinating, sowing the germinated seeds in a tray containing the matrix for growing seedlings, applying the mixture of acidithiobacillus ferrooxidans, Coriolus versicolor, and paracoccidentalis, leveling, improving the polluted soil in mining area, digging ditch, burying the seedlings in ditch, returning soil, covering seedlings, field management, growing for 90-120 days and harvesting. By the method, after one or more times of planting, the physicochemical indexes of the coal mining area soil can be obviously improved to reach the normal standard, a new way is provided for the soil treatment of the vast coal mining area, and the method has wide application prospect.
Description
Technical Field
The invention belongs to the technical field of soil bioremediation, and particularly relates to a method for restoring coal mining area soil.
Background
Coal has a high percentage in energy consumption in China all the time, and although coal has a descending trend in the percentage of primary energy consumption for five years continuously with the green development concept and the push of policies such as the active solution of excess capacity in the coal industry in 2016, and the percentage is reduced from 70.2% in 2010 to 62.3% in 2016, in the future, China still has an energy structure mainly based on coal. In the development and utilization process of coal, the ecological safety of a mining area is inevitably damaged, which is particularly shown in the aspects of soil pollution and damage, water pollution and damage, atmospheric pollution, noise pollution, biological resource damage, mining area ecological environment damage and the like, and the heavy metal pollution of the soil is one of the main aspects. Heavy metals in coal mine soil mainly originate from 3 aspects: (1) the dust generated in the coal mining process is migrated and settled, so that the coal mine dust is redistributed in the soil around the coal mine under the action of wind power and enters the soil through leaching and infiltration, thereby increasing the pollution to the surrounding soil; (2) the coal gangue produced by coal mining contains heavy metal with higher concentration, and the coal gangue enters soil along with surface runoff under the scouring and leaching action of atmospheric precipitation or is suspended in the atmosphere in a flying dust form through wind erosion in the stacking process of the coal gangue and finally falls into the soil around the coal gangue pile; (3) the coal mine acidic wastewater with high solubility and corrosivity on ores and surrounding rocks can aggravate the dissolution of heavy metals in the ores and the surrounding rocks, and carry a large amount of harmful chemical substances such as heavy metals and the like into a water body to pollute irrigation soil. For the coal producing provinces in China, such as Shanxi province, Shaanxi province, inner Mongolia autonomous region and other places, the environment and soil conditions of the surrounding areas of the coal mine are seriously affected, the soil has high heavy metal content and multiple types, the soil almost loses the production capacity, even if some soils can be used for planting crops, the crops contain a large amount of heavy metal components due to the infiltration of heavy metals, if the heavy metals enter human bodies through food chains, the harm to the health is unimaginable, and the life health of surrounding residents is greatly threatened. Therefore, the remediation and treatment of the coal mining area soil are imperative.
The existing coal mine area soil remediation and treatment methods mainly comprise physical and chemical remediation and biological remediation, and also comprise clay remediation in a research stage. The physical and chemical remediation mainly comprises the steps of leaching, chemical curing or electric remediation of soil, and the like, and the technologies not only have high cost and are not suitable for large-scale implementation, but also can reduce the activity of the soil, prevent the heavy metal pollution from spreading, but do not improve the soil recovery, so the application is very few. In recent years, bioremediation has been favored by researchers. The bioremediation technology is a process of improving the physicochemical properties of soil and improving the biological activity of soil by mainly utilizing the metabolic activities of organisms or microorganisms in a system, so that the ecological environment of the soil is partially or completely restored to the original state. Particularly, for repairing the soil in the coal mining area, plants and microorganisms with strong heavy metal absorption capacity are planted in the polluted area, so that the heavy metals are separated from the soil, the pollution is reduced, and the soil is recovered. Patent CN106607454A discloses a method for restoring chromium-contaminated soil in a coal mine subsidence area, which comprises the steps of firstly collecting soil samples by adopting a grid multi-point method, then measuring the chromium content of the collected soil samples, grading the soil according to the soil chromium content measurement result, then ploughing the soil on the surface layer of the chromium-contaminated soil, then spreading microbial inoculum and soil improvement particles with corresponding dosage according to the graded soil level, finally irrigating the chromium-contaminated soil, and keeping the relative humidity of the soil to be more than 85% for 10-20 days. The literature indicates that the method has high restoration efficiency, low cost and no secondary pollution, can restore the chromium-polluted soil in the subsidence area of the coal mine more accurately and quickly according to local conditions, effectively prevents chromium from migrating in the soil, provides nutrient elements for the soil for a long time in the soil restoration process and improves the available value of the soil. However, the method only adopts a single microbial remediation technology, and the remediation efficiency is not verified through experiments and lacks of persuasion. Moreover, the method is only effective for chromium in the soil heavy metal pollutants, other main heavy metal pollutants such as copper, lead, mercury, arsenic and the like cannot be effectively treated, and the soil pollution condition cannot be thoroughly improved, so that the practical application of the method is limited.
Disclosure of Invention
The invention aims to provide a method for restoring coal mining area soil, which utilizes the synergistic effect generated between two different types of bioremediation means to realize the restoring effect which cannot be achieved by a single bioremediation means, and can carry out biological absorption on various heavy metals, thereby thoroughly improving the soil quality and improving the utilization value of the coal mining area soil.
The technical scheme adopted by the invention is as follows: a method for repairing coal mining area soil by combining plants and microorganisms comprises the following steps:
step one, soaking the three-feather grass seeds with the water content of 12-15% in 1% by mass of lime water for 1-2 hours, fishing out, uniformly paving, covering with wet cotton gauze, and watering for 2-4 times every day at the temperature of 20-30 ℃, wherein the single watering amount is based on the condition that the cotton gauze can not hold water any more until the three-feather grass seeds germinate;
step two, planting the germinated three-hair grass seeds into seedling raising hole trays filled with seedling raising substrates, burying for 1.5-2cm, planting 2-3 seeds into each hole tray, keeping the temperature at 22-32 ℃, controlling the relative humidity of the seedling raising substrates at 65-85%, watering for 2-3 times every day, and controlling the single watering amount to be based on the water seeping from the bottoms of the hole trays until the seedlings of the three-hair grass are drilled out of the surface;
step three, after the seedling plants drill out of the surface, controlling the temperature of a seedling culture hole tray to be 25-35 ℃, controlling the relative humidity of a seedling culture substrate to be 60-80%, watering for 2-3 times every day, wherein the single watering amount is based on the water seepage from the bottom of the seedling culture hole tray, performing light supplement treatment by using blue light with the illumination intensity of 2200-;
step four, after the step three is completed, applying the mixed bacteria diluent to a seedling raising plug tray with seedlings, wherein the application amount is up to the time when the mixed bacteria diluent begins to seep out from the bottom of the seedling raising plug tray, and the mixed bacteria diluent is prepared by mixing acidithiobacillus ferrooxidans, coriolus versicolor thrombus fungus and paracoccidiobolus mould solid fungus agents according to the mass ratio of (5-7) to (2-3) to (1-2) to obtain a mixed fungus agent, and then diluting the mixed fungus agent by 100 times by using distilled water to obtain the mixed fungus diluent;
step five, transplanting the seedling plants applied with the mixed bacteria diluent into the polluted soil of the pretreated mining area, watering the seedling plants at irregular periods in the growth process to ensure that the water content of the soil is not less than 50% of the field water capacity, cutting off three-hairy grass stalks after the seedling plants grow for more than 30 cm, detecting whether the physicochemical indexes of the soil reach the standard, and repeatedly planting the seedling plants once or for many times if the physicochemical indexes of the soil do not reach the standard.
As a preferred mode: and the preparation process of the seedling raising substrate in the second step is that 25-35 parts of garden soil, 9-13 parts of humus, 4-6 parts of carbonized rice hulls and 3-5 parts of chicken manure are uniformly mixed according to the mass ratio, and then carbendazim is sprayed for disinfection, wherein the spraying amount of the carbendazim is 5 liters of 500 times of liquid sprayed on each 1 cubic meter.
As a preferred mode: in the fourth step, the solid microbial inoculum of the acidithiobacillus ferrooxidans, the coriolus versicolor trametes and the sacculus coronarius is 5:3:1 in mass ratio.
As a preferred mode: and step five, the polluted soil in the mining area is pretreated by flattening the polluted soil in the mining area, applying humic acid with the thickness of 2-3 cm on the surface of the soil, turning over and mixing the soil with the thickness of 30 cm below the surface layer of the soil and the humic acid uniformly, and digging ditches after 20 days, wherein the depth of each ditch is 0.25 m, and the row spacing is 0.6-0.8 m.
As a preferred mode: the transplanting process in the fifth step is that the seedling plants applied with the mixed bacteria diluent and the attached soil are transplanted in the ditch from the seedling culture hole tray, the covering thickness of the returned soil is 10-20 cm, and the planting density is 800-1200 plants per square meter.
The basic conditions of the plant and microorganism varieties involved in bioremediation of the present invention are as follows:
all-grass of three-wool grassTrisetumbifidum) The alias crab grass belongs to the gramineae, is a perennial grass plant, and grows beside hillside roads, in shade places and on ditch-edge wet grasslands with the elevation of 490-2500 meters. It is mostly seen in the provinces of Gansu, Tibet, Shaanxi, Henan, Jiangsu, Anhui, Zhejiang, Fujian, Jiangxi, Hubei, Hunan, Sichuan, Guizhou, Yunnan, Guangxi, Guangdong, etc. of China. Is suitable for growing on fertile loam or sandy loam and can resist acid soil. The seeds germinate in early spring, grow fast, bloom in 5 months, and fruit in 6 months, and the seeds are easy to fall off after being mature, and mainly propagate.
Acidithiobacillus ferrooxidans (A), (B), (C)Acidithiobacillusferrooxins) Using CO in the air for aerobic chemoautotrophic microorganisms2As a carbon source by oxidation of Fe in the matrix2+Is Fe3+And inorganic sulfur as sulfate to obtain energy. The characteristic makes the coal desulfurization agent have potential application value in the fields of bacterial metallurgy, acidic wastewater treatment, flue gas desulfurization, coal desulfurization and the like.
Trametes versicolor (trametes versicolor) ((Trametes versicolor) Belonging to the Basidiomycota family of wood-decay fungi, also known asCoriolus versicolor, coriolus versicolor mushroom, etc. The research shows that the compound contains steroids, triterpenes, organic acids, alkaloids, glucitol, proteins, saccharides, glycopeptides and other compounds. The strain is mainly used for health care products and is only applied to the aspects of wastewater treatment and soil nickel pollution treatment in the field of environmental protection.
Paracoronarum sacculus mildew (A)Glomus coronatum) Belongs to one of arbuscular mycorrhizal fungi, spores are singly grown in soil, and are spherical, hemispherical or irregular, and the diameter of the spores is 80-150 multiplied by 95-230 um. Orange to yellow-brown. The spore wall is two layers, the outer wall is a transparent wall, the thickness is 1.5-3.5 um, and the wall is easy to fall off along with the mature spore and is not easy to observe; the thickness of the yellow to brown inner wall can reach 4-6 um, the width of hypha connection points can reach 30-42 um, the hypha connection points are typically funnel-shaped, the connection positions of the hyphomycete and spores are separated by a bent partition, and the hyphomycete and the spores are researched and applied to the aspect of being used as plant bacterial manure at present.
The action mechanism of the invention for repairing the coal mining area soil is as follows:
the plant-microbe combined repairing technology is one pollution treating technology of degrading pollutant and eliminating environmental pollutant with the composite system comprising plant and microbe. The method utilizes the coexistence relationship of soil, microorganisms and plants, fully exerts the respective advantages of the plants and the microorganism restoration technology, makes up the defects, further improves the phytoremediation efficiency of pollutants in the soil, and finally achieves the aim of thoroughly restoring the soil. The microorganisms and the plants are combined to form symbiont, such as mycorrhiza, so that the absorption of heavy metals by plant root systems can be reduced, and the rapid planting of the plants is promoted, and therefore, the microbial symbiont is very necessary in the ecological restoration of mining areas.
Arbuscular mycorrhizal fungi which are beneficial symbiotic microorganisms commonly existing in the rhizosphere of terrestrial plants can form symbiota with 80% of plants on the land, and the mineral nutrient absorption capacity of the plants can be improved through a huge hypha network under the barren condition; under extreme environment, the heavy metal resistance, drought resistance, antibacterial capability and carbon sequestration capability of plants are enhanced through nutrition improvement, and finally the microbial environment of soil is greatly improved.
The three-feather grass has strong adaptability to the growing environment, can tolerate the acid soil environment, can absorb various heavy metals such as copper, zinc, lead, chromium, mercury and the like in the soil after being planted in the coal mining area soil, has a growing period of only about 3 months, can repeatedly grow and reap in a short period of time, and improves the soil remediation efficiency. The paracoccidiobolus in the mixed microbial inoculum can not only form symbiont with plants to improve the capability of the plants to absorb heavy metals, but also form stable micro-ecological environment together with acidithiobacillus ferrooxidans and coriolus versicolor trametes to decompose organic pollutants in soil. More importantly, the composite microbial inoculum formed by the three components is applied, so that the absorption efficiency and the tolerance capacity of the three-hairy grass to heavy metal can be further obviously improved compared with the case of only applying single arbuscular mycorrhizal fungi, the remediation effect of the polluted soil of the coal mining area is improved, and the technology has feasibility of practical popularization and application.
The invention has the beneficial effects that:
(1) according to the invention, field experiments are carried out on various herbaceous plants with short growing periods, so that the three-grass is determined as the repairing plant, and the expected repairing effect is realized. The three-feather grass is only applied to a few animals as livestock forage grass in previous researches, and the three-feather grass is applied to the first time of remediation of heavy metal soil.
(2) The invention finally determines the combination of acidithiobacillus ferrooxidans, coriolus versicolor trametes and paracoccidiobolus by screening and proportioning a plurality of microorganisms applied to soil remediation. The three bacteria play a differentiated role in the technical scheme of the invention, are matched with each other, and have a synergistic effect better than that of applying a single bacteria, and the microorganism not only can effectively promote the planting of the three-hair weeds and improve the capacity of the root systems of the three-hair weeds to absorb heavy metal ions, but also can activate heavy metals in soil and improve the biological effectiveness.
(3) The method realizes the field planting of the three-feather grasses by promoting the grass seeds to sprout, cultivating seedlings by using the seedling cultivation hole trays and finally burying the seedlings in the soil of the mining area for growth, and compared with the common method of directly sowing grass seeds in the soil, the method can greatly improve the survival rate of the three-feather grasses. And during seedling raising, the proportion of the seedling raising matrix is optimized, and light supplement treatment is carried out by time intervals and wavelength after seedling emergence, so that the growth and development of plant seedlings are promoted, the tolerance capacity of the seedlings after field planting in coal mining area soil is improved, and the growth condition of the seedlings after field planting is obviously improved.
(4) The invention applies humic acid to the soil of the coal mining area for reaction, thereby improving the physicochemical property of the soil and being beneficial to the field planting and the growth of the Chinese alpine rush seedlings. Reasonable planting density and field management also have non-negligible influence on the maximization of the final restoration effect.
Detailed Description
The present invention will be described in more detail with reference to examples.
The method comprises the following steps of (1) utilizing three-hairy-grass and mixed microbial inoculum to restore the soil of the coal mining area:
(1) spreading the three-feather grass seeds on the ground covered with a plastic film under the condition that the climate temperature is higher than 10 ℃, carrying out insolation by utilizing sunlight, wherein the total time of the insolation is 24 hours, then soaking the seeds by using lime water with the concentration of 1 percent, fishing out the grass seeds after soaking for 2 hours, uniformly spreading the grass seeds on sandy soil, covering the grass seeds by using wet cotton gauze, keeping the temperature at 25 ℃, watering for 3 times every day, and controlling the amount of water for one time based on the wet cotton gauze until the grass seeds germinate;
(2) preparing a seedling substrate according to the proportion of 25 parts of garden soil, 13 parts of humus, 5 parts of carbonized rice hulls and 4 parts of chicken manure, spraying carbendazim for disinfection, wherein 5L of 500 times of carbendazim is sprayed per 1 cubic meter of seedling substrate, then, containing the seedling substrate in a seedling hole tray, sowing germinated grass seeds in the seedling hole tray, controlling the seedling temperature to be 28 ℃ and the relative humidity of the substrate to be 80%, watering for 3 times every day, and preferably, a little water seeps from the bottom of the hole tray when the water is watered once until seedlings emerge;
(3) after seedling emergence, controlling the seedling raising temperature to be 28 ℃, controlling the relative humidity of a substrate to be 75%, watering for 3 times every day, wherein the amount of water for single watering is preferably that a little water seeps out of the bottom of a plug tray, and performing light supplement treatment by using blue light with the illumination intensity of 2200Lx at 3:30-5:30 every day, and performing light supplement treatment by using green light with the illumination intensity of 2000Lx at 21:30-23:30, when a seedling plant grows to 3 cm, removing redundant seedlings in the plug tray, ensuring that only one seedling is reserved in a single plug tray, performing seedling supplement treatment on the plug tray without seedling emergence, and simultaneously manually cleaning weeds;
(4) the source of the acidithiobacillus ferrooxidans, the coriolus versicolor trametes and the paracoccidiobolus are as follows: the acidithiobacillus ferrooxidans is purchased from China general microbiological culture Collection center (CGMCC), and the preservation number is CGMCC No. 3265; the Coriolus versicolor trametes is purchased from China general microbiological culture Collection center (CGMCC), and the preservation number is CGMCC No. 12241; the paracoccidiobolus paraguani is purchased from China arbuscular mycorrhizal fungi germplasm resource bank with the preservation number of BGC NM 06A.
When in use, preparing a paracoccidiobolus mould solid microbial inoculum in advance, taking sieved river sand as a substrate, sterilizing the substrate by high-pressure steam at the temperature of 121 ℃, placing the substrate for 8 days in a natural state, then putting 2 Kg of the substrate into a basin, inoculating 100g of original seeds of the paracoccidiobolus mould, adding tap water accounting for 25 percent of the weight of the substrate after uniformly stirring, sowing artemisia desertorum seeds, keeping normal growth, and obtaining host plants infected by the paracoccidiobolus mould and the substrate containing spores and extra-root hyphae of the paracoccidiobolus mould after five to six months; cutting off aerial parts of Artemisia desertorum, cutting into root segments, and mixing with substrate containing Paracoronarum sacculus mildew spores and extracellular hypha to obtain solid microbial inoculum of Paracoronarum sacculus mildew.
Inoculating acidithiobacillus ferrooxidans and trametes versicolor into different shake flask liquid culture medium, culturing at constant temperature in shake flask oscillator, collecting seed solution, adding into small fermentation tank containing different culture medium, and continuously culturing to increase viable count to 1 × 108One per ml. The culture medium for acidithiobacillus ferrooxidans comprises, per 1L: FeSO4·7H2O44.7g，(NH4)2SO43.0g，K2HPO40.5g，MgSO4·7H2O0.5g，KCl0.1g，Ca(NO3)20.01g, pH of the medium was 2. The culture medium for trametes versicolor comprises the following components in 1L: KH (Perkin Elmer)2PO41.0g，Na2HPO40.2g，MgSO4·7H2O 0.5g， VB1 0.1mg，CaCl20.1mg，FeSO4·7H2O 0.1mg，ZnSO4·7H2O 0.01mg，CuSO4·5H2O0.2 mg, glucose 1.0g, ammonium tartrate 0.1g, pH of the culture medium was 5.
Separating the acidithiobacillus ferrooxidans and trametes versicolor after amplification culture to obtain thalli, and mixing the thalli with a previously obtained solid microbial inoculum of sacculus coronarius according to the mass ratio of the acidithiobacillus ferrooxidans: trametes versicolor suppository: mixing paracoccidiobolus solid microbial inoculum =5:3:1 to obtain a mixed microbial inoculum, then diluting the mixed microbial inoculum by 100 times by using distilled water to obtain a mixed microbial inoculum, applying the mixed microbial inoculum to each hole with seedlings, and preferably, a little of the diluted microbial inoculum is leaked out of the bottom of a hole tray in a single application amount;
(5) selecting a coal mine in a certain country in Ningwu county of Xinzhou city, Shanxi province, defining a waste land with a test range within 5 kilometers of the central radius of a coal mine mining area, leveling the polluted soil in the mining area, and applying humic acid (humic acid raw powder is purchased from Shanxi Jinwohumic acid technology Limited company, the content of effective components is more than or equal to 50 percent of humic acid, more than or equal to 80 percent of organic matter, less than or equal to 20 percent of water and less than or equal to 25 percent of water) on the surface of the soil, wherein the thickness of the humic acid is 2.5 centimeters, then soil with the thickness of 30 cm below the surface layer of the soil and humic acid are turned and mixed evenly to react for 20 days, then ditches are dug according to the row spacing of 0.7 m, the depth of the ditches is 0.25 m, the three-hair grass seedlings applied with the mixed bacteria diluent and the attached soil are taken out from the holes of the seedling culture hole tray and buried in the ditches, then returning soil to cover the seedlings, wherein the thickness of the returned soil covering is 15 cm, and the planting density is 1000 plants per square meter;
(6) and (3) performing field management on the planted three-grass, watering at irregular intervals according to the actual condition of soil moisture to ensure that the soil moisture is not less than 50% of the field moisture capacity, and harvesting after the three-grass grows for 100 days.
Meanwhile, the polluted land in the circled test range is restored by planting the three-hairy grass according to the completely same test conditions, the difference is that only a single glomus coronarius is used for replacing the mixed microbial inoculum in the step (4), and other parameters related to strain expansion culture and microbial inoculum application are not changed and are used as a comparison example of plant and single microbial inoculum restoration.
The method comprises the following steps of (1) soil heavy metal content change determination and three-grass reaped heavy metal adsorption capacity determination:
the method for measuring the content of the heavy metal in the soil comprises the following steps: removing impurities such as broken stones, plant residual roots and the like in the soil sample, air-drying, and sieving by a 100-mesh sieve for later use. By HNO3:HF:HClO4(v: v =4:4:2), and the contents of copper, zinc, lead, chromium, mercury, and arsenic were measured with a proeutectoid TAS-900 atomic absorption spectrophotometer.
The method for measuring the heavy metal content in the harvested Trifolium pratense plant bodies comprises the following steps: deactivating enzyme of herba Trifolii Pratentis plant and root system at 105 deg.C for 30 min, oven drying at 75 deg.C to constant weight, pulverizing, and sieving with 100 mesh sieve. Followed by HClO4:HNO3(v: v =2: 5), and the contents of copper, zinc, lead, chromium, mercury, and arsenic were measured with a proeutectoid TAS-900 atomic absorption spectrophotometer.
By using the above measuring method, the contents of heavy metals of copper, zinc, lead, chromium, mercury and arsenic in original soil in a coal mine test range of a country in Ningwu county, Xinzhou province, Shanxi province are respectively measured (sample 1), the contents of heavy metals of copper, zinc, lead, chromium, mercury and arsenic in soil after a period of three-haired grass restoration (sample 2), the contents of heavy metals of copper, zinc, lead, chromium, mercury and arsenic in three-haired grass harvested after a restoration period is completed (sample 3), and the contents of heavy metals of copper, zinc, lead, chromium, mercury and arsenic in three-haired grass harvested after a restoration period and a single microbial inoculum restoration control example (sample 4), wherein each group of samples is in parallel, and each test index is shown in tables 1-4.
TABLE 1 content (mg/Kg) of heavy metals copper, zinc, lead, chromium, mercury and arsenic in original soil
TABLE 2 content (mg/Kg) of heavy metals copper, zinc, lead, chromium, mercury, arsenic in soil after one cycle of Trifolium grass restoration
TABLE 3 content (mg/Kg) of heavy metals of copper, zinc, lead, chromium, mercury and arsenic in the harvested Trifolium grass after completion of one restoration cycle
TABLE 4 contents (mg/Kg) of heavy metals Cu, Zn, Pb, Cr, Hg and As in the harvested plants after completion of a restoration cycle and in the three-grass of the comparative example with a single microbial inoculum restoration
The experimental results show that the three-feather grasses serving as the coal mining area soil remediation plants can obviously reduce the content indexes of the heavy metals of copper, zinc, lead, chromium, mercury and arsenic which exceed the national grade I standard of soil environmental quality in the soil in one growth period, and the content of the heavy metal elements except lead is completely reduced to be below the standard maximum limit value, so that a good remediation effect is obtained. Moreover, compared with the control example only applying the paracoccidiobolus dose diluting solution, the adsorption capacity of the three-hairy-grass applied with the mixed bacteria diluting solution to the heavy metals of copper, zinc, lead, chromium, mercury and arsenic in the growth period of the three-hairy-grass is obviously improved, and the mixed bacteria diluting solution plays a non-negligible role in the adsorption capacity of the heavy metals of the three-hairy-grass, so that the soil bioremediation method has a prospect of popularization and implementation.
Although the present invention has been described in detail in the examples by way of general illustration, specific embodiments and experiments, it is to be understood that modifications and improvements may be made without departing from the spirit of the invention and within the scope of the following claims.
Claims (5)
1. A method for repairing coal mining area soil by combining plants and microorganisms is characterized by comprising the following steps: the method comprises the following steps:
step one, soaking the three-feather grass seeds with the water content of 12-15% in 1% by mass of lime water for 1-2 hours, fishing out, uniformly paving, covering with wet cotton gauze, and watering for 2-4 times every day at the temperature of 20-30 ℃, wherein the single watering amount is based on the condition that the cotton gauze can not hold water any more until the three-feather grass seeds germinate;
step two, planting the germinated three-hair grass seeds into seedling raising hole trays filled with seedling raising substrates, burying for 1.5-2cm, planting 2-3 seeds into each hole tray, keeping the temperature at 22-32 ℃, controlling the relative humidity of the seedling raising substrates at 65-85%, watering for 2-3 times every day, and controlling the single watering amount to be based on the water seeping from the bottoms of the hole trays until the seedlings of the three-hair grass are drilled out of the surface;
step three, after the seedling plants drill out of the surface, controlling the temperature of a seedling culture hole tray to be 25-35 ℃, controlling the relative humidity of a seedling culture substrate to be 60-80%, watering for 2-3 times every day, wherein the single watering amount is based on the water seepage from the bottom of the seedling culture hole tray, performing light supplement treatment by using blue light with the illumination intensity of 2200-;
step four, after the step three is completed, applying the mixed bacteria diluent to a seedling raising plug tray with seedlings, wherein the application amount is up to the time when the mixed bacteria diluent begins to seep out from the bottom of the seedling raising plug tray, and the mixed bacteria diluent is prepared by mixing acidithiobacillus ferrooxidans, coriolus versicolor thrombus fungus and paracoccidiobolus mould solid fungus agents according to the mass ratio of (5-7) to (2-3) to (1-2) to obtain a mixed fungus agent, and then diluting the mixed fungus agent by 100 times by using distilled water to obtain the mixed fungus diluent;
step five, transplanting the seedling plants applied with the mixed bacteria diluent into the polluted soil of the pretreated mining area, watering the seedling plants at irregular periods in the growth process to ensure that the water content of the soil is not less than 50% of the field water capacity, cutting off three-hairy grass stalks after the seedling plants grow for more than 30 cm, detecting whether the physicochemical indexes of the soil reach the standard, and repeatedly planting the seedling plants once or for many times if the physicochemical indexes of the soil do not reach the standard.
2. The method for repairing the soil of the coal mining area by combining the plants and the microorganisms according to claim 1, wherein the method comprises the following steps: and the preparation process of the seedling raising substrate in the second step is that 25-35 parts of garden soil, 9-13 parts of humus, 4-6 parts of carbonized rice hulls and 3-5 parts of chicken manure are uniformly mixed according to the mass ratio, and then carbendazim is sprayed for disinfection, wherein the spraying amount of the carbendazim is 5 liters of 500 times of liquid sprayed on each 1 cubic meter.
3. The method for repairing the soil of the coal mining area by combining the plants and the microorganisms according to claim 1, wherein the method comprises the following steps: in the fourth step, the solid microbial inoculum of the acidithiobacillus ferrooxidans, the coriolus versicolor trametes and the sacculus coronarius is 5:3:1 in mass ratio.
4. The method for repairing the soil of the coal mining area by combining the plants and the microorganisms according to claim 1, wherein the method comprises the following steps: and step five, the polluted soil in the mining area is pretreated by flattening the polluted soil in the mining area, applying humic acid with the thickness of 2-3 cm on the surface of the soil, turning over and mixing the soil with the thickness of 30 cm below the surface layer of the soil and the humic acid uniformly, and digging ditches after 20 days, wherein the depth of each ditch is 0.25 m, and the row spacing is 0.6-0.8 m.
5. The method for repairing the soil of the coal mining area by combining the plants and the microorganisms according to claim 4, wherein the method comprises the following steps: the transplanting process in the fifth step is that the seedling plants applied with the mixed bacteria diluent and the attached soil are transplanted in the ditch from the seedling culture hole tray, the covering thickness of the returned soil is 10-20 cm, and the planting density is 800-1200 plants per square meter.
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