CN108265010B - Method for repairing cadmium-polluted soil by using aspergillus versicolor and Indian mustard - Google Patents
Method for repairing cadmium-polluted soil by using aspergillus versicolor and Indian mustard Download PDFInfo
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Abstract
The invention relates to a method for repairing cadmium contaminated soil by using aspergillus versicolor and Indian mustard, belonging to the technical field of microorganisms. The Aspergillus sydowii (Aspergillus sydowii) of the invention has been deposited in the China general microbiological culture Collection center (CGMCC) No. 02/05 in 2018, and the location is No. 3 of Xilu No.1 of Beijing Kogyo-Yang area, the deposition number is CGMCC No. 15385. The invention provides a method for repairing cadmium-polluted soil by using aspergillus versicolor and Indian mustard, which has the advantages of economy, high efficiency, no secondary pollution, landscape beautification and the like.
Description
Technical Field
The invention relates to a method for repairing cadmium contaminated soil by using aspergillus versicolor and Indian mustard, belonging to the technical field of microorganisms.
Background
Recently, with the rapid development of industry and agriculture, heavy metals are widely used, and the pollution problem is more and more obvious. Soil is used as a place for accommodating the largest heavy metal pollutants, so that the heavy metal pollution of the soil is one of global environmental problems. Therefore, the heavy metal pollution of soil, especially the farmland soil, is in urgent need of prevention and repair.
The cadmium in the soil is mainly from the mining and smelting of lead and cadmium ores and the discharge of three wastes in the electroplating industry. At present, the area of the cultivated land in China polluted by heavy metals such as cadmium and cadmium accounts for 1/5 of the total area of the cultivated land. Excessive cadmium in soil can cause soil degradation, influence plant growth and cause crop yield reduction, cadmium is accumulated in human bodies through food chains after being enriched by plants, the cadmium harms human health, causes digestive tract symptoms such as human vomiting, diarrhea, anorexia and the like, and causes other symptoms such as human anemia and the like.
Therefore, the soil quality is the first line of defense for the safety production of agricultural products, and the prevention, control and repair of heavy metal pollution in soil are of great significance. However, heavy metal contamination in soil has latency, irreversibility, longevity, and universality, and cannot be removed by biodegradation, and only site migration and interconversion between various forms occur.
At present, the remediation method for the heavy metal contaminated soil mainly comprises three methods, namely a physical method, a chemical method and a biological method. The traditional physical and chemical remediation method realizes the removal or passivation of heavy metals in soil through various physical processes or by adding modifiers, inhibitors and the like into the soil, and comprises a soil-moving method, soil leaching, electric remediation, electric-heating remediation, stabilization/solidification and the like, and the technologies have the defects of high cost, damage to the soil structure, easy reduction of soil fertility and possible secondary pollution.
In recent years, much research on bioremediation technology is paid much attention due to the characteristics of low investment, low cost, small disturbance to soil environment, no secondary pollution and the like, wherein the method of absorbing heavy metal by using plants is considered to be one of ideal methods for solving heavy metal pollution of soil, and the Indian mustard is one of super-accumulator plants, can absorb, enrich and transfer heavy metal cadmium, and is a plant resource with good remediation capability.
However, the plants which have been found to be useful for soil bioremediation are limited by geographical and climatic conditions, generally have a low biomass and grow slowly, and thus greatly affect their potential for remediation of heavy metal contaminated soil.
Disclosure of Invention
The invention provides a method for repairing cadmium-polluted soil by using aspergillus versicolor and Indian mustard, which has the advantages of economy, high efficiency, no secondary pollution, landscape beautification and the like. According to the invention, the Indian mustard is planted in the cadmium-polluted soil, and Aspergillus sydowii (Aspergillus sydowii) capable of repairing the cadmium-polluted soil is inoculated on the root of the Indian mustard, so that heavy metal cadmium is enriched in the Indian mustard, and the Indian mustard is integrally removed after the Indian mustard is mature, thereby achieving the purpose of removing the heavy metal cadmium pollutant in the soil.
The technical scheme of the invention is as follows:
the invention provides an Aspergillus sydowii (Aspergillus sydowii) for repairing cadmium-polluted soil, which is preserved in the China general microbiological culture Collection center (CCCEC) at No. 02-05 of 2018, with the address of No. 3 Siro-1 of Beijing city Kogyo-Chen-Yang area and the preservation number of CGMCC No. 15385.
The invention provides application of Aspergillus sydowii (Aspergillus sydowii) for repairing cadmium-polluted soil in treating cadmium pollution.
The invention provides a method for repairing cadmium contaminated soil by using aspergillus versicolor, which comprises the following steps:
step 1: planting Indian mustard in the cadmium-polluted soil;
step 2: inoculating cadmium-resistant microorganisms to the root system of Indian mustard;
and step 3: repeating the steps 1 and 2 after a batch of Indian mustard grows to be mature until the cadmium content in the soil is lower than 0.2 mg/kg;
the cadmium-resistant microorganism is Aspergillus sydowii (Aspergillus sydowii); the Aspergillus sydowii (Aspergillus sydowii) is deposited in the China general microbiological culture Collection center at No. 02/05 of 2018, with the address of No. 3 Siro-1 of Beijing, Chaoyang, and the deposition number of CGMCC No. 15385.
In one embodiment of the invention, the Indian mustard is planted in cadmium contaminated soil, and the water content of the soil is controlled to be kept between 55 and 65 percent.
In one embodiment of the invention, the Indian mustard is planted in cadmium contaminated soil in an open air cultivation mode.
In one embodiment of the invention, the inoculation is carried out at a density of not less than 1X 10 at the root system of Brassica juncea6CFU/g of Aspergillus polytrichus.
In one embodiment of the invention, the inoculation is a preparation of Aspergillus polytrichus at a concentration of 1-4X 108CFU/mL of the bacterial suspension, and spraying the bacterial suspension respectively before sowing the Indian mustard and at 2, 4 and 6 weeks after emergence.
In one embodiment of the invention, the inoculation is a preparation of Aspergillus polytrichus at a concentration of 1-4X 108CFU/mL of bacterial suspension, 5mL of bacterial suspension is sprayed on the 2 nd, 4 th and 6 th weeks before sowing and after emergence of Indian mustard.
In one embodiment of the invention, the spraying of the bacterial suspension is spraying the bacterial suspension onto the surface of the Indian mustard.
The invention provides an application of the method for repairing cadmium-polluted soil by using aspergillus versicolor and Indian mustard in the aspect of treating cadmium pollution.
Has the advantages that:
(1) the Aspergillus sydowii has high cadmium tolerance and Cd2+The Minimum Inhibitory Concentration (MIC) for the bacterium is 1500 mg/L;
(2) the Aspergillus sydowii provided by the invention can promote the increase of the biomass of the India mustard, and the fresh weight and the dry weight of the overground part of the India mustard can be obviously increased by 14-125% and 7-134% and the fresh weight and the dry weight of the underground part of the India mustard can be obviously increased by 12-63% and 21-82% after the Aspergillus sydowi is inoculated to the root system of the India mustard;
(3) the method provided by the invention is used for repairing cadmium-contaminated soil, the enrichment and absorption of Indian mustard on heavy metal cadmium in the soil can be remarkably promoted, the extraction effect of the Indian mustard on the cadmium is improved, after the bacteria are inoculated to the root system of the Indian mustard, the concentration of the effective cadmium in the soil can be improved by 36 percent at most, meanwhile, the concentration of the cadmium on the overground part of the Indian mustard is improved by 4 to 36 percent, and the concentration of the cadmium on the underground part is improved by 23 to 36 percent;
(4) the method for repairing cadmium-polluted soil provided by the invention has the advantages of strong practicality, high efficiency, safety, no damage to soil structure and property, no secondary pollution, environment beautification and the like, and has wide application prospect.
Biological material preservation
A species of Aspergillus sydowii (Aspergillus sydowii) classified under the name: aspergillus polytrichus, the preservation unit is: the China general microbiological culture Collection center has the following preservation addresses: xilu No.1 Hospital No. 3, Beijing, Chaoyang, with the deposit number: CGMCC No.15385, the preservation date is: year 2018, month 02, day 05.
Drawings
FIGS. 1-4 Effect of the strains of the invention on Indian mustard biomass at various cadmium concentrations.
FIG. 5 Effect of the strains of the present invention on the available cadmium concentration in soil at different cadmium concentrations.
FIGS. 6-7 Effect of inoculation with the strains of the invention on cadmium concentration in Brassica oleracea.
FIG. 8 Effect of inoculation with the strains of the invention on Total cadmium uptake by Indian mustard.
Detailed Description
The following examples are provided as further illustration of the invention and are not to be construed as limitations or limitations of the invention.
The detection method of the Indian mustard biomass comprises the following steps:
mixing and sampling each pot of plants, shearing the plants along the soil surface, dividing the plants into an overground part and an underground part, washing the plants with distilled water, drying the plants with absorbent paper, weighing the fresh weight with an electronic balance, then placing the plants in a drying oven at 105 ℃ for fixation for 0.5h, drying the plants to a constant weight at 65 ℃, and weighing the dry weight with the electronic balance.
The detection method of the content of the available cadmium in the soil comprises the following steps:
after the potted plant is sampled, a soil sample around the original rhizosphere is dug, the soil sample is dried by air and then passes through a 60-mesh nylon sieve, 0.1mol/LHCl extractant is added according to the soil-liquid ratio of 1:10, and the content of the effective cadmium in the soil is measured by using a flame atomic absorption spectrophotometer. The detection method of cadmium absorption amount of Indian mustard comprises the following steps:
crushing the dried samples, taking 0.2g of each sample, adding mixed acid of nitric acid and perchloric acid (4:1, v: v), performing microwave digestion, and measuring the Cd content of the overground and underground parts by using a flame atomic absorption spectrophotometer.
The method for detecting the total cadmium absorption amount of the Indian mustard comprises the following steps:
the cadmium content of each part of the plant is multiplied by the dry weight of each part of the plant and then added to obtain the total cadmium absorption amount.
The strains in the following examples are Aspergillus sydowii (Aspergillus sydowii) screened from cadmium-contaminated soil, which has been deposited in the general microbiological center of China Committee for culture Collection of microorganisms No. 05 in 2018, having the accession number CGMCC No. 3, CGMCC No.15385, in North Kyowa, sunny region, Beijing.
Examples
The specific implementation steps are as follows:
(1) adding CdCl 0, 0.0856, 0.1713 and 0.3425g into soil2Adding water into the solid powder, stirring and uniformly mixing to ensure that the concentration of the metal Cd in the soil is respectively 0, 28, 56 and 112mg/kg, setting 3 potted plants for each concentration, and finally taking an average value to simulate Cd-polluted soil;
(2) sterilizing at 121 deg.C for 20min after soil stabilization, cooling, and placing into pots, each containing 1.5kg of Cd contaminated soil, 6 pots being set at each concentration as inoculation group and control group, and 24 pots in total;
(3) respectively sowing Indian mustard seeds in a direct seeding mode in the inoculation group and the control group, sowing 8 seeds in each pot, thinning the seedlings 1 week after the seeds germinate, and reserving 5 seedlings with good growth vigor in each pot;
(4) 5mL of the solution with the concentration of 1-4 multiplied by 10 is sprayed on the 2 nd, 4 th and 6 th weeks before sowing and after emergence of the inoculated group potted plant8CFU/mL suspension of Aspergillus sydowii (Aspergillus sydowii) was added to each pot of the control groupEqual amount of sterile water;
(5) during the growth period of the plants, natural ventilation is kept, enough illumination is provided, watering is carried out in the morning and at night every day, the soil is ensured to be moist, the water content of the soil is controlled to be kept between 55 and 65 percent, and the plants are harvested after the Indian mustard is mature.
The results of the Indian mustard biomass measurements were as follows:
the effect of the Aspergillus polytrichus of the present invention on the biomass of Brassica juncea at different soil Cd concentrations can be seen in FIGS. 1-4. As can be seen from fig. 1 and 2: compared with the non-inoculated group, the overground fresh weights of the inoculated group Indian mustard in the soil with Cd concentrations of 0, 28, 56 and 112mg/kg respectively rise by 125%, 122%, 41% and 14%, and the underground fresh weights respectively rise by 25%, 54%, 12% and 63%. As can be seen from fig. 3 and 4: the dry weight of the overground part of the inoculated group plants in the soil with Cd concentrations of 0, 28, 56 and 112mg/kg is respectively increased by 134%, 115%, 39% and 7% compared with the non-inoculated group, and the dry weight of the underground part is respectively increased by 52%, 82%, 31% and 21%.
The results show that compared with the control group, the Aspergillus versicolor increases the quality of each part of the Indian mustard, and particularly has more remarkable promoting effect in the soil with the concentration of 0 mg/kgCd and the soil with the concentration of 28 mg/kgCd.
The detection result of the content of the available cadmium in the soil is as follows:
the influence of inoculating the aspergillus polyticus bacterium on the cadmium content of the effective state of the soil under different soil cadmium concentrations can be shown in figure 5. As shown in fig. 5: at metal concentrations of 28, 56 and 112mg/kg, the soil effective cadmium concentration of the inoculation group is respectively increased by 36 percent, 27 percent and 4 percent compared with that of the control group.
The metal elements in the soil exist in different chemical forms, and the metal elements in different chemical forms can be enriched by plants only after being converted into effective states. The content of the metal in the effective state can evaluate the environmental risk, the biological effectiveness and the toxicity of the polluted soil, so that the soil pollution condition can be known by researching the effective state of the metal. In the embodiment, the content of the soil effective cadmium is increased by inoculating the aspergillus polytrichus of the invention, and under the treatment of 28mg/kg cadmium, the concentration of the soil effective cadmium of an inoculated group is increased by 36% at most compared with that of a non-inoculated group, which shows that the aspergillus polytrichus of the invention can dissolve the insoluble cadmium in the soil and increase the content of the soil effective Cd.
The detection result of the cadmium absorption amount of the Indian mustard is as follows:
the effect of inoculating Aspergillus polytrichus of the present invention at different cadmium concentrations on cadmium uptake by Brassica juncea can be seen in FIGS. 6-7. As shown in fig. 6: when the Cd concentrations in the soil are 28, 56 and 112mg/kg, compared with the control group, the Cd concentrations in the overground part of the Indian mustard of the inoculation group are respectively increased by 4 percent, 16 percent and 26 percent; as shown in fig. 7: when the Cd concentration of the soil is 28, 56 and 112mg/kg, the Cd concentration of the underground part of the Indian mustard of the inoculation group is obviously improved by 33 percent, 36 percent and 23 percent compared with that of the control group.
In the experiment, the Cd concentrations of the overground part and the underground part of the India mustard inoculated with the Aspergillus polytrichus are increased to different degrees, which is probably because the Aspergillus polytrichus promotes the dissolution of the insoluble cadmium in the soil, increases the concentration of the effective Cd in the soil and improves the absorption and enrichment of the India mustard on the cadmium.
The detection result of the total cadmium absorption of the Indian mustard is as follows:
the effect of inoculating the strains of the invention on total cadmium uptake by Indian mustard is shown in FIG. 8. As shown in fig. 8: in the soil with different cadmium concentrations, the total cadmium absorption of the Indian mustard inoculated with the strain is respectively improved by 122%, 108%, 31% and 46% compared with that of the non-inoculated group, which shows that the strain can obviously improve the total cadmium absorption of the Indian mustard.
Although the present invention has been described with reference to the preferred embodiments, it should be understood that various changes and modifications can be made therein by those skilled in the art without departing from the spirit and scope of the invention as defined in the appended claims.
Claims (21)
1. The method for repairing cadmium contaminated soil by using aspergillus versicolor in combination with brassica juncea is characterized by comprising the following steps:
step 1: planting Indian mustard in the cadmium-polluted soil;
step 2: inoculating cadmium-resistant microorganisms to the root system of Indian mustard;
and step 3: repeating the steps 1 and 2 after a batch of Indian mustard grows to be mature until the cadmium content in the soil is lower than 0.2 mg/kg;
the cadmium-resistant microorganism is Aspergillus sydowii (Aspergillus sydowii); the Aspergillus sydowii (Aspergillus sydowii) is deposited in the China general microbiological culture Collection center at No. 02/05 of 2018, with the address of No. 3 Siro-1 of Beijing, Chaoyang, and the deposition number of CGMCC No. 15385.
2. The method for remediating cadmium-contaminated soil using aspergillus versicolor in combination with brassica juncea as claimed in claim 1, wherein the brassica juncea is planted in the cadmium-contaminated soil, and the water content of the soil is controlled to be maintained at 55-65%.
3. The method for remediating cadmium-contaminated soil using aspergillus versicolor in combination with brassica juncea as claimed in claim 2, wherein the brassica juncea is planted in the cadmium-contaminated soil in an open air cultivation manner.
4. The method for remediating cadmium-contaminated soil using aspergillus versicolor in combination with brassica juncea as claimed in claim 1, wherein the inoculation is performed by inoculating the brassica juncea root system with a density of not less than 1 x 106CFU/g of Aspergillus polytrichus.
5. The method for remediating cadmium-contaminated soil using aspergillus versicolor in combination with brassica juncea as claimed in claim 2, wherein the inoculation is performed by inoculating the brassica juncea root system with a density of not less than 1 x 106CFU/g of Aspergillus polytrichus.
6. The method for remediating cadmium-contaminated soil using Aspergillus versicolor in combination with Brassica juncea as claimed in claim 3, wherein the inoculation is carried out by inoculating the root system of Brassica juncea at a density of not less than 1 x 106CFU/g of Aspergillus polytrichus.
7. The method for remediating cadmium contaminated soil using Aspergillus sydowii in combination with Brassica juncea as claimed in claim 1, wherein said inoculating is to prepare Aspergillus sydowii to a concentration of 1-4 x 108CFU/mL of the bacterial suspension, and spraying the bacterial suspension respectively before sowing the Indian mustard and at 2, 4 and 6 weeks after emergence.
8. The method for remediating cadmium contaminated soil using Aspergillus sydowii in combination with Brassica juncea as claimed in claim 2, wherein said inoculating is to prepare Aspergillus sydowii to a concentration of 1-4 x 108CFU/mL of the bacterial suspension, and spraying the bacterial suspension respectively before sowing the Indian mustard and at 2, 4 and 6 weeks after emergence.
9. The method for remediating cadmium contaminated soil using Aspergillus sydowii in combination with Brassica juncea as claimed in claim 3, wherein said inoculating is to prepare Aspergillus sydowii to a concentration of 1-4 x 108CFU/mL of the bacterial suspension, and spraying the bacterial suspension respectively before sowing the Indian mustard and at 2, 4 and 6 weeks after emergence.
10. The method for remediating cadmium contaminated soil using Aspergillus sydowii in combination with Brassica juncea as claimed in claim 4, wherein said inoculating is to prepare Aspergillus sydowii to a concentration of 1-4 x 108CFU/mL of the bacterial suspension, and spraying the bacterial suspension respectively before sowing the Indian mustard and at 2, 4 and 6 weeks after emergence.
11. The method for remediating cadmium contaminated soil using Aspergillus sydowii in combination with Brassica juncea as claimed in claim 5, wherein said inoculating is to prepare Aspergillus sydowii to a concentration of 1-4 x 108CFU/mL of the bacterial suspension, and spraying the bacterial suspension respectively before sowing the Indian mustard and at 2, 4 and 6 weeks after emergence.
12. The method for remedying cadmium pollution by using Aspergillus sydowii in combination with Brassica juncea as claimed in claim 6The soil inoculation method is characterized in that the Aspergillus sydowii is prepared into a concentration of 1-4 multiplied by 108CFU/mL of the bacterial suspension, and spraying the bacterial suspension respectively before sowing the Indian mustard and at 2, 4 and 6 weeks after emergence.
13. The method for remediating cadmium contaminated soil using Aspergillus sydowii in combination with Brassica juncea as claimed in claim 1, wherein said inoculating is to prepare Aspergillus sydowii to a concentration of 1-4 x 108CFU/mL of bacterial suspension, 5mL of bacterial suspension is sprayed on the 2 nd, 4 th and 6 th weeks before sowing and after emergence of Indian mustard.
14. The method for remediating cadmium contaminated soil using Aspergillus sydowii in combination with Brassica juncea as claimed in claim 2, wherein said inoculating is to prepare Aspergillus sydowii to a concentration of 1-4 x 108CFU/mL of bacterial suspension, 5mL of bacterial suspension is sprayed on the 2 nd, 4 th and 6 th weeks before sowing and after emergence of Indian mustard.
15. The method for remediating cadmium contaminated soil using Aspergillus sydowii in combination with Brassica juncea as claimed in claim 3, wherein said inoculating is to prepare Aspergillus sydowii to a concentration of 1-4 x 108CFU/mL of bacterial suspension, 5mL of bacterial suspension is sprayed on the 2 nd, 4 th and 6 th weeks before sowing and after emergence of Indian mustard.
16. The method for remediating cadmium contaminated soil using Aspergillus sydowii in combination with Brassica juncea as claimed in claim 4, wherein said inoculating is to prepare Aspergillus sydowii to a concentration of 1-4 x 108CFU/mL of bacterial suspension, 5mL of bacterial suspension is sprayed on the 2 nd, 4 th and 6 th weeks before sowing and after emergence of Indian mustard.
17. The method for remediating cadmium contaminated soil using Aspergillus sydowii in combination with Brassica juncea as claimed in claim 5, wherein said inoculating is to prepare Aspergillus sydowii to a concentration of 1-4 x 108CFU/mL of bacterial suspension, 5mL of bacterial suspension is sprayed on the 2 nd, 4 th and 6 th weeks before sowing and after emergence of Indian mustard.
18. The method for remediating cadmium contaminated soil using Aspergillus sydowii in combination with Brassica juncea as claimed in claim 6, wherein said inoculating is to prepare Aspergillus sydowii to a concentration of 1-4 x 108CFU/mL of bacterial suspension, 5mL of bacterial suspension is sprayed on the 2 nd, 4 th and 6 th weeks before sowing and after emergence of Indian mustard.
19. The method for remediating cadmium contaminated soil using Aspergillus sydowii in combination with Brassica juncea as claimed in claim 7, wherein said inoculating is to prepare Aspergillus sydowii to a concentration of 1-4 x 108CFU/mL of bacterial suspension, 5mL of bacterial suspension is sprayed on the 2 nd, 4 th and 6 th weeks before sowing and after emergence of Indian mustard.
20. The method for remediating cadmium contaminated soil using Aspergillus sydowii in combination with Brassica juncea as claimed in claim 7, wherein the spraying the bacterial suspension is spraying the bacterial suspension onto the surface of Brassica juncea.
21. The method for remediating cadmium contaminated soil using aspergillus versicolor in combination with brassica juncea as claimed in claim 13, wherein the spraying the bacterial suspension is spraying the bacterial suspension onto the surface of brassica juncea.
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