CN110756571A - Method for adsorbing heavy metals in soil by combining plants and microorganisms - Google Patents
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- CN110756571A CN110756571A CN201911059772.1A CN201911059772A CN110756571A CN 110756571 A CN110756571 A CN 110756571A CN 201911059772 A CN201911059772 A CN 201911059772A CN 110756571 A CN110756571 A CN 110756571A
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Abstract
The invention relates to the technical field of soil remediation, and discloses a method for adsorbing heavy metals in soil by combining plants and microorganisms. The invention relates to a plant-microorganism combined repair technology which takes artemisia alba as a plant raw material and VA fungi as a microorganism raw material. Collecting partial soil with the depth of 30cm under the artemisia selengensis, and sieving to collect the exposed VA fungal spores. Adding mixture of river mud and soil into beef extract-peptone culture medium, sterilizing, and sterilizing. Inoculating the obtained VA fungal spore into a culture medium for culturing. Sterilizing Artemisia ordosica seed with hydrogen peroxide solution, and soaking in warm water. Inoculating the obtained VA fungi into heavy metal-containing soil, performing multi-layer inoculation to ensure the inoculation amount of spores, and then putting the treated Artemisia ordosicae seeds. The VA fungus is formed along with the growth of the artemisia alba. Compared with the existing plant-microorganism combined repair technology, the Cu adsorption effect is better and can reach 85%.
Description
Technical Field
The invention relates to the technical field of soil remediation, in particular to a method for adsorbing heavy metals in soil by combining plants and microorganisms.
Background
At present, the soil problem gradually shows harm, particularly the problem that the content of heavy metal in the soil exceeds the standard is more serious, and the exceeding of the heavy metal in the soil can cause a lot of harm. Heavy metals cannot be degraded by microorganisms, so that heavy metal substances can be retained in soil for a long time and are difficult to treat, and the heavy metal substances can influence the development of plant roots and leaves and infect underground water after being retained in the soil for a long time, so that the living health of human beings is influenced. However, the prior treatment technology for heavy metals in soil is time-consuming, labor-consuming and high in cost, and the method for adsorbing the heavy metals in the soil by using arbuscular fungi (VA fungi) and plants is low in cost and has obvious treatment effect on copper and lead.
Disclosure of Invention
In order to overcome the defects, the invention provides a technical measure for effectively treating the heavy metals of copper and lead in the soil, which can more effectively remove the heavy metals in the soil to reach the content specified by the state. The technical scheme adopted for realizing the purpose of the invention is as follows: a method for adsorbing heavy metals in soil by combining plants and microorganisms comprises the following steps,
1) collecting VA fungal spores: selecting tall and healthy artemisia alba to dig downwards, collecting soil at a position 30cm away from the ground, screening by using a 100-mesh sieve, and collecting exposed VA fungal spores;
2) preparation of a culture medium: adding a mixture of river mud and soil into a beef extract-peptone culture medium, uniformly mixing, placing into a 121 ℃ high-temperature steam sterilization pot for sterilization for 15-20min, and sterilizing with 20% hydrogen peroxide solution for 15min to obtain a culture medium;
3) culture of VA fungi: inoculating VA fungal spores obtained in the step 1) into the culture medium obtained in the step 2), and culturing in a climatic chamber for 3 days at the temperature of 30 ℃ to culture VA fungi;
4) pretreating artemisia selengensis seeds: putting the seeds of the artemisia alba into a hydrogen peroxide solution with the mass fraction of 30% for disinfection for 15min, and then putting the seeds into warm water with the temperature of 45 ℃ for soaking for 3 days;
5) inoculating the cultured VA fungi obtained in the step 3) into soil containing heavy metals, and then inoculating the artemisia alba seeds pretreated in the step 4) into the soil containing the VA fungi and the heavy metals.
In the method for adsorbing heavy metals in soil by combining plants and microorganisms, in the step 2), the preparation method of the beef extract-peptone culture medium comprises the following steps: taking 3g of beef extract; 10g of peptone; 5g of sodium chloride; 15g of agar; 1000ml of water was prepared at a pH of 7.4-7.6.
In the method for the combined adsorption of heavy metals in soil by the plant and the microorganism, in the step 2), the weight ratio of beef extract to peptone medium is as follows: mixture of river mud and soil 1: 1-5.
In the method for adsorbing heavy metals in soil by combining plants and microorganisms, in the step 3), according to the mass ratio, VA fungal spores: medium 1: 15-20.
In the method for adsorbing heavy metals in soil by combining plants and microorganisms, in the step 3), the inoculation mode is a plate coating method.
In the method for adsorbing the heavy metal in the soil by combining the plants and the microorganisms, the inoculation amount of the VA fungus is 0.8-1g/m 2.
In the method for adsorbing heavy metals in soil by combining plants and microorganisms, the inoculation amount of the artemisia alba seeds is 15-20g/m 2.
The method for adsorbing the heavy metal in the soil by the combination of the plants and the microorganisms is characterized in that the heavy metal is copper or lead.
The invention has the beneficial effects that:
(1) the plant-microorganism of the invention has very obvious effect on adsorbing copper and lead heavy metals in soil.
(2) The preparation process of the plant-microorganism of the invention is relatively simple, and the raw materials have wide sources and low price. The method has less investment for removing the heavy metals in the soil and better obtained effect.
(3) The plant-microorganism of the invention can not only adsorb the heavy metals of copper and lead in soil, but also beautify the environment.
Drawings
FIG. 1 shows the effect of the amount of plant-microorganism microorganisms used in the examples.
FIG. 2 shows the effect of the plant-microorganism effect under different pH conditions in the examples.
FIG. 3 is a graph showing the effect of the amount of arbuscular fungus on the absorption of heavy metals in soil.
Detailed Description
Example 1 method for the Combined plant-microorganism adsorption of heavy metals in soil
1. Collecting VA fungal spores:
1) the tall and healthy Artemisia ordosica is dug under the selected trees, and 50g of soil is collected at the position which is 30cm deep from the ground.
2) The collected soil was sieved through a 100 mesh sieve.
3) Collecting the exposed VA fungal spores.
2. Preparation of the culture Medium
1) Preparation of beef extract peptone medium: taking 3g of beef extract; 10g of peptone; 5g of sodium chloride; 15g of agar; 1000ml of water was prepared at a pH of 7.4 to 7.6.
2) 10g of river mud and soil were added to 50g of beef extract peptone medium.
3) Placing the culture medium obtained in the step 2) into a high-temperature steam sterilization pot at 121 ℃ for sterilization for 15-20 min. Then, 15g of hydrogen peroxide solution with the mass fraction of 20% is used for fumigation for 15 min.
3. Culture of VA fungi:
inoculating the obtained arbuscular fungal spores onto a culture medium by using a plate coating method, putting the culture medium into an artificial climate box for culturing for 3 days, controlling the temperature at 30 ℃, and culturing the VA fungi.
4. Pretreating artemisia selengensis seeds:
1) taking 20g of Artemisia ordosica seed, placing into 50ml of 30% hydrogen peroxide solution, and sterilizing for 15 min.
2) Soaking the disinfected artemisia alba seeds obtained in the step 1) in warm water at 45 ℃ for 3 days. Obtaining the pretreated artemisia alba seeds.
5. Method for adsorbing heavy metals in soil by combining plants and microorganisms
According to the inoculation amount of 0.8-1g/m2Inoculating VA fungi into soil with excessive copper content according to the inoculation amount of 15-20g/m2Inoculating Artemisia ordosica seed to the soil, and adsorbing for 30 days under neutral pH condition. And measuring the content of copper in the soil before and after adsorption.
Then, a soil heavy metal analyzer is used, soil with the excessive copper which is put into the flowerpot at first is used as reference, the heavy metal content is respectively determined, and the adsorption rate and the adsorption quantity of the plant-microorganism to the copper are calculated according to the following formula:
in the formula: q-amount of copper adsorbed by plant-microorganism (mg);
c0-initial content of copper in soil (mg);
c-content of copper (mg) in soil after 30 days;
v is the mass (g) of adsorbed copper;
through experimental determination, the plant-microorganism can adsorb heavy metal copper in soil, and the copper content before adsorption is 600 mg/kg; the copper content after adsorption is 90mg/kg, and the adsorption rate reaches 85 percent.
(II) determination of optimal conditions of plant-microorganism in process of adsorbing heavy metal
1. Effect of different amounts of arbuscular fungi
(1) 750g of soil with the copper content of 600mg/kg is added into 8 flowerpots respectively, and the reference numerals are given as 1, 2, 3, 4, 5, 6, 7 and 8.
(2) Sequentially adding 2, 4, 6, 8, 10, 12, 14 and 16mg of VA fungi into the eight flowerpots respectively. Then 2.8g of the seeds of the artemisia alba are put in the seeds for cultivation for 30 days.
(3) And finally, respectively measuring the adsorption capacity by using a soil heavy metal analyzer and taking the soil with the copper content of 600mg/kg which is added into the flowerpot at the beginning as a reference.
As a result, as shown in FIG. 1, the absorption amount increased with the increase in the amount of the arbuscular fungus, and it is understood from FIG. 1 that the absorption amount reached the maximum when the mass of the arbuscular fungus charged was 14 mg. Therefore, for 750g of soil with 600mg/kg of copper content exceeding the standard, the optimal input amount of the arbuscular fungi is 14 mg.
2. Effect of different pH:
(1) the pH meter was adjusted to a pH of 5.24 and a pH of 9.18 in a buffer solution, and adjusted to the state required for the experiment according to the procedure on the pH meter using the instructions. The original soil was found to have a pH of 6.84.
(2) 750g of soil with a copper content of 600mg/kg were added to 5 pots and the pH of the solution was preconditioned with a 1:9 strength sulfuric acid solution and a 1:5 sodium hydroxide solution (original soil pH 6.84). The soil with pH values of 3, 5, 7, 9 and 11 is obtained. Then 14mg of arbuscular fungi are added into the soil, 2.8g of artemisia alba seeds are planted, and cultivation is carried out for 30 days.
(3) And finally, respectively measuring the adsorption capacity by using a soil metal analyzer and taking the soil with the copper content of 600mg/kg which is added into the flowerpot at the beginning as a reference. The copper content before adsorption is 600 mg/kg; the copper content after adsorption was 90 mg/kg.
As shown in FIG. 2, the adsorption effect of the plant-microorganism increases first and then decreases as the pH increases. The maximum adsorption rate at neutral was 85%. I.e. the pH is 7, the effect is optimal.
3. Separate use of VA fungus to adsorb heavy metal in soil
(1) 750g of soil with the copper content of 600mg/kg is added into 8 flowerpots and is marked with the numbers 1, 2, 3, 4, 5, 6, 7 and 8.
(2) Sequentially adding 2, 4, 6, 8, 10, 12, 14 and 16mg of VA fungi into the eight flowerpots respectively. The culture was carried out for 30 days.
(3) And finally, respectively measuring the adsorption capacity by using a soil heavy metal analyzer and taking the soil with the copper content of 600mg/kg which is added into the flowerpot at the beginning as a reference.
As shown in FIG. 3, the absorption increased with the increase of the amount of arbuscular mycorrhizal fungi, and for 750g of soil with a copper content of 600mg/kg, the absorption rate was 60.34% at the maximum when the amount of VA fungi added was 14mg, but the absorption rate was not as good as when it was used in combination with Artemisia alba.
4. Separate use of artemisia selengensis for adsorbing heavy metals in soil
(1) 750g of soil with the copper content of 600mg/kg is added into the flowerpot.
(2) And then 2.8g of artemisia alba seeds are put into the pot to be cultivated for 30 days.
(3) And finally, respectively measuring the adsorption capacity by using a soil heavy metal analyzer and taking the soil with the copper content of 600mg/kg which is added into the flowerpot at the beginning as a reference.
The final result is that the heavy metal content in the soil is reduced but the effect is not as obvious as that of the mixed use of plant-microorganism under the condition of singly using the artemisia alba. The content of copper in the soil after 30 days of adsorption by the artemisia alba is 450 mg/kg.
Claims (8)
1. A method for adsorbing heavy metals in soil by combining plants and microorganisms is characterized by comprising the following steps,
1) collecting VA fungal spores: selecting tall and healthy artemisia alba to dig downwards, collecting soil at a position 30cm away from the ground, screening by using a 100-mesh sieve, and collecting exposed VA fungal spores;
2) preparation of a culture medium: adding a mixture of river mud and soil into a beef extract-peptone culture medium, uniformly mixing, placing into a 121 ℃ high-temperature steam sterilization pot for sterilization for 15-20min, and sterilizing with 20% hydrogen peroxide solution for 15min to obtain a culture medium;
3) culture of VA fungi: inoculating VA fungal spores obtained in the step 1) into the culture medium obtained in the step 2), and culturing in a climatic chamber for 3 days at the temperature of 30 ℃ to culture VA fungi;
4) pretreating artemisia selengensis seeds: putting the seeds of the artemisia alba into a hydrogen peroxide solution with the mass fraction of 30% for disinfection for 15min, and then putting the seeds into warm water with the temperature of 45 ℃ for soaking for 3 days;
5) inoculating the cultured VA fungi obtained in the step 3) into heavy metal soil for one layer per 1cm of thickness, inoculating 5-8 layers, and then inoculating the artemisia alba seeds pretreated in the step 4) into the soil containing the VA fungi and the heavy metals.
2. The method for plant-microorganism combined adsorption of heavy metals in soil according to claim 1, wherein in step 2), the beef extract-peptone medium is prepared as follows: taking 3g of beef extract; 10g of peptone; 5g of sodium chloride; 15g of agar; 1000ml of water was mixed at pH 7.4-7.6 to obtain beef extract-peptone medium.
3. The method for the combined plant-microorganism adsorption of heavy metals in soil according to claim 1 or 2, wherein in the step 2), the weight ratio of beef extract-peptone medium: mixture of river mud and soil 1: 1-5.
4. The method for the combined plant-microorganism adsorption of heavy metals in soil according to claim 1, wherein in the step 3), the mass ratio of VA fungal spores: medium 1: 15-20.
5. The method for the combined plant-microorganism adsorption of heavy metals in soil according to claim 4, wherein in step 3), the inoculation method is a plate coating method.
6. The method for plant-microorganism combined adsorption of heavy metals in soil according to claim 1, wherein the inoculation amount of VA fungi is 0.8-1g/m2。
7. The method for plant-microorganism combined adsorption of heavy metals in soil according to claim 1, wherein the inoculation amount of artemisia alba seeds is 15-20g/m2。
8. The method according to claim 1, wherein the heavy metal is copper or lead.
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| CN111530920A (en) * | 2020-04-30 | 2020-08-14 | 辽宁大学 | Method for jointly repairing petroleum-polluted soil by using surfactant-enhanced plant microorganisms |
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Application publication date: 20200207 |