CN112387778A - Method for restoring heavy metal contaminated soil by combining sludge biochar immobilized bacteria with plants - Google Patents
Method for restoring heavy metal contaminated soil by combining sludge biochar immobilized bacteria with plants Download PDFInfo
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- CN112387778A CN112387778A CN202011088518.7A CN202011088518A CN112387778A CN 112387778 A CN112387778 A CN 112387778A CN 202011088518 A CN202011088518 A CN 202011088518A CN 112387778 A CN112387778 A CN 112387778A
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- 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
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- 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
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Abstract
The invention relates to a method for repairing heavy metal contaminated soil by combining sludge biochar immobilized bacteria and plants, which comprises the steps of screening plants, screening and separating bacteria with resistance to various heavy metals, separating bacteria capable of dissolving phosphorus, fixing nitrogen, secreting plant growth hormone, secreting iron carriers, promoting plant biomass accumulation, root development and growth hormone secretion, fixing bacterial load on the sludge biochar, repairing soil and the like. The invention fixes the bacterial strain by using the sludge biochar, the microorganism is protected by the biochar and can be slowly released in soil, the survival and the planting capability of the microorganism in the plant body and at the rhizosphere are effectively improved, the biomass of the plant is greatly improved, the heavy metal content of the soil is effectively reduced, the root extraction of the restoration plant is promoted, and the aim of restoring the heavy metal pollution of the soil by combining the plant-microorganism-carbon-based material is realized.
Description
Technical Field
The invention relates to the field of heavy metal soil remediation, in particular to a method for remediating heavy metal contaminated soil by combining sludge biochar immobilized bacteria and plants.
Background
At present, a plurality of bioremediation technologies are applied to heavy metal contaminated soil in mining areas, wherein a method for achieving a synergistic effect by utilizing plant-microorganism combined remediation is widely accepted. However, microorganisms artificially added at present and having a repair function have various problems of difficulty in survival, low repair efficiency, difficulty in competition with indigenous microorganisms, easiness in functional degradation and the like in the environment.
Disclosure of Invention
Based on the above disadvantages of the prior art, it is necessary to provide a method for repairing heavy metal contaminated soil by combining sludge biochar immobilized bacteria and plants, wherein the bacterial strains are immobilized by using the sludge biochar, so that the survival and planting abilities of microorganisms in plant bodies and rhizosphere are improved, the heavy metal content in the soil is reduced, and the heavy metal extraction of the repaired plants is promoted.
In order to achieve the above purpose, the solution adopted by the invention is as follows: a method for restoring heavy metal contaminated soil by combining sludge biochar immobilized bacteria and plants comprises the following steps:
s1, screening out plants with high resistance level to heavy metals;
s2, screening and separating bacteria with resistance to various heavy metals from the mining area restoration plants;
s3, screening the plant growth promoting and soil improvement performance of the bacteria which are obtained in the step S2 and have resistance to various heavy metals to obtain a single bacterial strain which can dissolve phosphorus, fix nitrogen, secrete plant growth hormone and secrete siderophore;
s4, performing a plant loop-back test on the bacteria obtained in the step S3 to obtain bacteria capable of promoting plant biomass accumulation, root development and growth hormone secretion;
s5, inoculating and mixing the sludge biochar with the fermentation liquor of the bacteria screened in the step S4, and culturing for 8-10 hours in a shaking table to ensure that the bacteria are loaded and fixed on the sludge biochar;
s6, planting the plants screened and obtained in the step S1 in an area needing soil remediation, and adding sludge biochar loaded with bacteria to the rhizosphere of the planted plants.
Further, the plants obtained by screening in step S1 are woody plants with high resistance to heavy metals.
Further, the plant obtained by screening in step S1 is robinia pseudoacacia having a high level of resistance to heavy metal cadmium.
Further, the bacteria separated in step S2 are rhizobacteria.
Further, the plant growth hormone described in step S3 includes IAA and ACC.
Further, in the step S5, the sludge biochar is mixed with the bacterial fermentation liquor according to the weight ratio of 10: 1.
Further, in step S6, sludge biochar is added to the rhizosphere of the plant by root irrigation.
The invention has the beneficial effects that:
(1) the bacterial strain is fixed by the sludge biochar, and the microorganism is protected by the biochar and can be slowly released in soil, so that the survival and planting capacity of the microorganism in the plant body and at the rhizosphere are effectively improved, the biomass of the plant is greatly improved, the heavy metal content of the soil is effectively reduced, and the heavy metal extraction of the restoration plant is promoted;
(2) according to the invention, a mode of fixing microorganisms by using biochar is utilized, so that the biochar and the microorganisms form a synergistic effect, the sludge biochar can be used for immobilizing heavy metals in soil, simultaneously plant extraction and overground part transfer of more heavy metals are promoted, the heavy metal stock in the soil can be removed while the biotoxicity of the heavy metals in the soil is effectively reduced, and the heavy metals are transferred to a part of a repairing plant, so that the recovery is convenient;
(3) the biochar provides trace element nutrition for plant stress resistance, such as P, K, provides a platform and a carrier for microorganism slow release and development, plays a role in immobilizing the occurrence form of soil heavy metal (effective state reduction), and obviously reduces the soil heavy metal through combined remediation.
Drawings
FIG. 1 is an electron micrograph of a bacterial load immobilized on a sludge biochar;
FIG. 2 is a statistical plot of the average cadmium content in the soil after potting experiments;
FIG. 3 is a statistical plot of the average exchangeable cadmium content in soil after potting experiments;
FIG. 4 is a statistical plot of the mean rhizosphere pH of plants after potting experiments;
FIG. 5 is a statistical chart of the average total length of roots of plants after potting experiment;
FIG. 6 is a statistical plot of the mean total surface area of the roots of plants after potting experiments.
Detailed Description
To facilitate an understanding of the invention, the invention will now be described more fully with reference to the accompanying drawings. Preferred embodiments of the present invention are shown in the drawings. This invention may, however, be embodied in many different forms and should not be construed as limited to the embodiments set forth herein. Rather, these embodiments are provided so that this disclosure will be thorough and complete.
Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. The terminology used herein in the description of the invention is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention. As used herein, the term "and/or" includes any and all combinations of one or more of the associated listed items.
Example 1: a method for restoring heavy metal contaminated soil by combining sludge biochar immobilized bacteria and plants comprises the following steps:
s1, screening the locust tree with high resistance level to heavy metal cadmium;
s2, screening and separating the locust rhizosphere bacteria which have resistance to various heavy metals from the mining area restoration plants, wherein the locust rhizosphere bacteria can be added into the rhizosphere of various plants including locust and the like to play a role;
s3, screening the bacteria which are obtained in the step S2 and have resistance to various heavy metals, and separating out a bacterial single strain which can dissolve phosphorus, fix nitrogen, secrete plant growth hormone (IAA and ACC) and secrete siderophores;
s4, performing a plant loop-back test on the bacteria obtained in the step S3 by using a pot experiment, and obtaining the bacteria capable of promoting plant biomass accumulation, root development and growth hormone secretion by regulating and controlling factors such as an inoculation mode, time, concentration and the like;
s5, sieving the sludge biochar, inoculating and mixing the sludge biochar and the bacterial fermentation liquor obtained by screening in the step S4 according to the weight ratio of 12:1, culturing for 8 hours at 25 ℃ in a shaking table to fix the bacterial load on the sludge biochar, and filtering, extracting and eluting the mixture of the sludge biochar and the bacterial fermentation liquor;
s6, planting the plants screened and obtained in the step S1 in areas (such as heavy metal contaminated tailings ponds and mining areas) needing soil remediation, irrigating roots of the plants according to the proportion of 0.05:1(w: w) of additive/rhizosphere soil, and adding sludge biochar (shown in figure 1) loaded with bacteria to rhizospheres of the plants.
Example 2: a method for restoring heavy metal contaminated soil by combining sludge biochar immobilized bacteria and plants comprises the following steps:
s1, screening the locust tree with high resistance level to heavy metal cadmium;
s2, screening and separating the locust rhizosphere bacteria which have resistance to various heavy metals from the mining area restoration plants, wherein the locust rhizosphere bacteria can be added into the rhizosphere of various plants including locust and the like to play a role;
s3, screening the bacteria which are obtained in the step S2 and have resistance to various heavy metals, and separating out a bacterial single strain which can dissolve phosphorus, fix nitrogen, secrete plant growth hormone (IAA and ACC) and secrete siderophores;
s4, performing a plant back grafting test on the bacteria obtained in the step S3 by using a pot experiment to obtain bacteria capable of promoting plant biomass accumulation, root development and growth hormone secretion;
s5, sieving the sludge biochar, inoculating and mixing the sludge biochar and the bacterial fermentation liquor obtained by screening in the step S4 according to the weight ratio of 10:1, culturing for 10 hours at 25 ℃ in a shaking table to fix the bacterial load on the sludge biochar, and filtering, extracting and eluting the mixture of the sludge biochar and the bacterial fermentation liquor;
s6, planting the plants screened and obtained in the step S1 in an area needing soil remediation, irrigating roots of the plants according to the proportion of 0.05:1(w: w) of the additive/rhizosphere soil, and adding sludge biochar loaded with bacteria to the rhizosphere of the plants.
Example 3: a method for restoring heavy metal contaminated soil by combining sludge biochar immobilized bacteria and plants comprises the following steps:
s1, screening the locust tree with high resistance level to heavy metal cadmium;
s2, screening and separating the locust rhizosphere bacteria which have resistance to various heavy metals from the mining area restoration plants, wherein the locust rhizosphere bacteria can be added into the rhizosphere of various plants including locust and the like to play a role;
s3, screening the bacteria which are obtained in the step S2 and have resistance to various heavy metals, and separating out a bacterial single strain which can dissolve phosphorus, fix nitrogen, secrete plant growth hormone (IAA and ACC) and secrete siderophores;
s4, performing a plant back grafting test on the bacteria obtained in the step S3 by using a pot experiment to obtain bacteria capable of promoting plant biomass accumulation, root development and growth hormone secretion;
s5, sieving the sludge biochar, inoculating and mixing the sludge biochar and the bacterial fermentation liquor obtained by screening in the step S4 according to the weight ratio of 8:1, culturing for 9 hours at 25 ℃ in a shaking table to fix the bacterial load on the sludge biochar, and filtering, extracting and eluting the mixture of the sludge biochar and the bacterial fermentation liquor;
s6, planting the plants screened and obtained in the step S1 in an area needing soil remediation, irrigating roots of the plants according to the proportion of 0.05:1(w: w) of the additive/rhizosphere soil, and adding sludge biochar loaded with bacteria to the rhizosphere of the plants.
Example 4:
according to the above method, sludge biochar loaded with bacteria was added to the rhizosphere of acacia, and then potted plant comparative experiments were performed. The potting matrix is Danish peat soil, the concentration of cadmium in the experimental soil is 60mg/kg, and the biochar is sludge biochar purchased from Anhui Tongyuan environment company. The experimental results are shown in fig. 2 to 6.
CK in the figure represents a control group without adding sludge biochar and bacteria; biochar represents an experimental group to which only sludge Biochar is added, Ch-8 represents an experimental group to which only screened locust rhizosphere bacteria are added, and B-Ch-8 represents an experimental group to which sludge Biochar loaded with locust rhizosphere bacteria is added. Each experimental group and the control group are provided with a plurality of potted plants, and other interference factors in the control group and the experimental group are almost the same, so that the experimental result is not influenced.
The locust rhizosphere bacteria can chelate Cd by secreting organic acid, siderophore and the like to form a biological defense mechanism under stress, and has the function of passivating Cd in soil. The sewage peat can mineralize heavy metals through the elements of phosphorus, iron and silicon which are rich in the sewage peat, and physically adsorbs cadmium through a pore structure, so that the pH value of the root system is increased. Therefore, the results of the Biochar experimental group, the Ch-8 experimental group and the B-Ch-8 experimental group show that the total Cd in the soil is obviously reduced, the bioavailability is obviously reduced, and the roots of the plants are enriched with more heavy metals but cannot be transported to the overground part compared with the CK control group. The treated plant roots can be enriched with more heavy metals, the sludge biochar has the most obvious growth promoting effect on improving the pH of soil and promoting the enrichment of the plant roots, and the B-Ch-8 experimental group has the most obvious effect on improving the microenvironment of the soil and has significance.
The test results of the above examples 1, 2, 3 and 4 show that the total amount of heavy metal cadmium in the soil is reduced by more than 79.3% and exchangeable cadmium (with biotoxicity) in the soil is reduced by more than 76% after 3 months of remediation by immobilizing the rhizosphere microorganisms of the robinia pseudoacacia through the sludge biochar and combining the microorganisms with the plant robinia pseudoacacia for remediation. The results of measurements on Robinia pseudoacacia plants show that example 1, example 2 and example 3, example 4 all increase Robinia pseudoacacia biomass and the root system is more developed (based on parameters such as main root length, root area and lateral root number).
The method provided by the invention promotes the plants to absorb heavy metals (underground roots, branches and leaves on the ground), plays a role in extracting the heavy metals by the plants, can be used for subsequently recovering the heavy metals, avoids secondary pollution caused by entering soil, and avoids heavy metals from flowing into a food chain again by extracting the heavy metals by inedible woody plants.
The technical features of the above embodiments can be arbitrarily combined, and for the sake of brevity, all possible combinations of the technical features in the above embodiments are not described, but should be considered as the scope of the present specification as long as there is no contradiction between the combinations of the technical features.
The above examples only show some embodiments of the present invention, and the description thereof is more specific and detailed, but not construed as limiting the scope of the invention. It should be noted that, for a person skilled in the art, several variations and modifications can be made without departing from the inventive concept, which falls within the scope of the present invention. Therefore, the protection scope of the present patent shall be subject to the appended claims.
Claims (7)
1. A method for restoring heavy metal contaminated soil by combining sludge biochar immobilized bacteria and plants is characterized by comprising the following steps:
s1, screening out plants with high resistance level to heavy metals;
s2, screening and separating bacteria with resistance to various heavy metals from the mining area restoration plants;
s3, screening the plant growth promoting and soil improvement performance of the bacteria which are obtained in the step S2 and have resistance to various heavy metals to obtain a single bacterial strain which can dissolve phosphorus, fix nitrogen, secrete plant growth hormone and secrete siderophore;
s4, performing a plant loop-back test on the bacteria obtained in the step S3 to obtain bacteria capable of promoting plant biomass accumulation, root development and growth hormone secretion;
s5, inoculating and mixing the sludge biochar with the fermentation liquor of the bacteria screened in the step S4, and culturing for 8-10 hours in a shaking table to ensure that the bacteria are loaded and fixed on the sludge biochar;
s6, planting the plants screened and obtained in the step S1 in an area needing soil remediation, and adding sludge biochar loaded with bacteria to the rhizosphere of the planted plants.
2. The method for remediating heavy metal contaminated soil by using sludge biochar immobilized bacteria and plants as claimed in claim 1, which is characterized in that: the plants screened in step S1 are heavy metal highly resistant woody plants.
3. The method for remediating heavy metal contaminated soil by using sludge biochar immobilized bacteria and plants as claimed in claim 2, which is characterized in that: the plant selected in step S1 is robinia pseudoacacia having a high level of resistance to heavy metal cadmium.
4. The method for remediating heavy metal contaminated soil by using sludge biochar immobilized bacteria and plants as claimed in claim 3, which is characterized in that: the bacteria separated in step S2 are rhizobacteria.
5. The method for remediating heavy metal contaminated soil by using sludge biochar immobilized bacteria and plants as claimed in claim 4, which is characterized in that: the plant growth hormones described in step S3 include IAA and ACC.
6. The method for remediating heavy metal contaminated soil by using sludge biochar immobilized bacteria and plants as claimed in claim 5, which is characterized in that: in the step S5, the sludge biochar is mixed with the bacterial fermentation liquor according to the weight ratio of 10: 1.
7. The method for remediating heavy metal contaminated soil by using sludge biochar immobilized bacteria and plants as claimed in claim 6, which is characterized in that: and in the step S6, adding sludge biochar to the rhizosphere of the plant in a root irrigation mode.
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