CN117903813A - Microbial soil activation microbial agent and preparation method and application thereof - Google Patents
Microbial soil activation microbial agent and preparation method and application thereof Download PDFInfo
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- CN117903813A CN117903813A CN202410288031.5A CN202410288031A CN117903813A CN 117903813 A CN117903813 A CN 117903813A CN 202410288031 A CN202410288031 A CN 202410288031A CN 117903813 A CN117903813 A CN 117903813A
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- azotobacter
- trichoderma harzianum
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Landscapes
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The invention discloses a microbial soil activation microbial agent, a preparation method and application thereof, and belongs to the technical field of microbial materials. The microbial agent comprises functional microbial fermentation liquid, a microbial activator and a microbial carrier, wherein the mass ratio of the functional microbial fermentation liquid to the microbial activator to the microbial carrier is 1:0.5:5. The trichoderma harzianum strain obtained by screening has extracellular adsorption and metal complexation effects, realizes reduction and discharge of metal ions by regulating expression of reductase and related genes, has higher arsenic ion removal efficiency, has capabilities of phosphate dissolving, nitrogen fixing and IAA production, and has the effects of regulating soil and promoting crop growth. After the trichoderma harzianum, the azotobacter chroococcus and the pseudomonas fragi are combined in proportion, the interaction is obvious, the heavy metal removal efficiency can be improved, and the synergistic growth is promoted. The three microorganisms can also regulate the content of nutrient substances in the soil, particularly regulate the activities of various enzymes in the soil, restore the ecological activity of the soil and have remarkable social and economic benefits.
Description
Technical Field
The invention belongs to the technical field of microbial materials, and particularly relates to a microbial soil activating microbial agent, a preparation method and application thereof.
Background
Heavy metals refer to certain biologically toxic metals and metalloids such as cadmium, mercury, arsenic, lead, chromium and the like. At present, the problem of heavy metal pollution of soil is increasingly focused. In urban heavy metal contaminated soil, enzymes and microorganisms are inhibited or even killed, which severely damages the ecological chain of the soil, affects the biological activity of the soil, and thus limits crop growth.
The treatment of the heavy metal pollution of the soil can be realized by adopting physical, chemical or biological treatment means. The physical and chemical means are used for repairing the soil polluted by heavy metals, so that the effect is poor, and the risk of secondary pollution to the soil is also caused. At present, heavy metals in soil are efficiently dissolved by microorganisms, so that the absorption of the heavy metal elements by plants is promoted; the rhizosphere microorganism is different from other microorganisms, and can secrete a large amount of plant hormone while dissolving heavy metals, so that the production and development of plants are promoted, and the repair efficiency of the plants to the heavy metals is improved.
However, at present, microorganisms are used for treating heavy metal elements in soil, repairing multiple metal elements in a plurality of ways, and the method has the advantages of single function and complex strains, and is difficult to effectively utilize and implement. For example: patent application number CN201610252293.1 discloses a microbial agent for improving soil heavy metals, comprising photosynthetic bacteria, actinomycetes, lactobacillus, saccharomycetes, bacillus subtilis, bacillus thuringiensis, bacillus megaterium, azotobacter, citric acid bacteria, bacillus licheniformis and a matrix for providing nutrition for the flora, and is used for improving soil heavy metals. The application number is CN202311153948.6, which discloses a microorganism composite microbial agent for repairing heavy metal contaminated soil, wherein each 100 parts of microorganism composite microbial agent comprises the following components in parts by weight: 5 parts of cellulomonas faecalis, 10 parts of pseudomonas nitroreduction, 30 parts of peptone, 35 parts of yeast extract and 20 parts of sodium chloride.
Therefore, how to develop a high-efficiency microbial soil activating microbial agent, and to activate soil environment and improve ecological activity and soil fertility while removing heavy metal elements in soil with high efficiency is a technical problem to be solved in urgent need at present.
Disclosure of Invention
Aiming at the problems existing in the prior art, the invention optimizes and separates a trichoderma harzianum strain, and forms functional microorganism strains with screened azotobacter chroococcus and pseudomonas meracilis, and can realize the efficient removal of heavy metal elements in soil under the combined action of a microorganism activator and a carrier, thereby having the functions of promoting growth and regulating soil.
In order to achieve the technical purpose, the invention adopts the following technical scheme:
a microbial soil activation microbial agent comprises functional microbial fermentation liquor, a microbial activation agent and a microbial carrier, wherein the mass ratio of the functional microbial fermentation liquor to the microbial activation agent to the microbial carrier is 1:0.5:5.
Preferably, the microbial activator is obtained by mixing sodium alginate and glucose according to a mass ratio of 1 (0.5-1.5).
Preferably, the microbial carrier is obtained by mixing (5-8)/(3-5)/(3-6)/(1-3) by mass ratio of bran, diatomite, biomass charcoal and fish bone powder.
Preferably, the functional microbial fermentation broth comprises trichoderma harzianum (Trichoderma harzianum), azotobacter chroococcus (Azotobacter chroococcum) and pseudomonas fragi (Pseudomonas plecoglossicida).
Preferably, the trichoderma harzianum (Trichoderma harzianum) has a preservation number of CGMCC No.40702, is preserved in the China general microbiological culture Collection center (China Committee for culture Collection of microorganisms), has a preservation date of 2023, 6 months and 16 days, and has a preservation address of beichen Xiyang No. 1, 3 of Beijing.
The trichoderma harzianum is separated from iron tailings in the mountain county of Jining, shandong province, and the content of various heavy metals including arsenic and lead in a sampling place exceeds the standard.
The separation method comprises the following steps: taking 5g of soil sample, placing the soil sample in a sterilized PDA culture medium, adding filtered and sterilized sodium arsenite solution, enabling the concentration of As 3+ to be 50mg/L, transferring the culture medium to a culture medium with the concentration of As 5+ of 100mg/L in an inoculum size of 2% after enrichment culture for 24 hours, culturing the culture medium under the same condition, gradually increasing the concentration of As 5+, carrying out dilution coating after transferring for 5 times, and separating and preserving the strain with the best growth vigor, namely the target strain trichoderma harzianum.
Morphological characteristics of the strain: colonies were grown on PDA plates at 25-28℃for 4 days for 5cm expansion, dark grey green; the mycelium had a septum and branches. Conidiophores form conifer-type branched outlines, and conidiophores are nearly spherical, oval, green and smooth in wall.
The nitrogen fixing bacteria have a preservation number of CGMCC No.1.5803, are purchased from the China general microbiological culture Collection center, and have an original preservation date of 6 months and 27 days in 2005; the Pseudomonas fragi has a preservation number of CGMCC No.1.16111, is purchased from the China general microbiological culture Collection center (China Committee for culture Collection of microorganisms), and has an original preservation date of 2017, 3 and 29. The azotobacter chroococcus and the pseudomonas fragi can be purchased through a collection center strain catalog, and additional collection is not needed.
The preparation method of the microbial soil activation microbial agent comprises the following preparation steps:
(1) Thawing Trichoderma harzianum, azotobacter chroococcus and Pseudomonas fragi, and respectively inoculating the Trichoderma harzianum, the azotobacter chroococcus and the Pseudomonas fragi in a PDA culture medium for activating culture; after activation culture, picking bacterial cakes of each bacterial strain, respectively adding the bacterial cakes into a mixed fermentation liquid culture medium according to an inoculation amount of 1.5% for fermentation culture, stopping fermentation when the effective viable count in each mixed fermentation liquid culture medium reaches 4X 10 8 CFU/mL to obtain three fermentation bacterial solutions, and mixing the three fermentation bacterial solutions according to a volume ratio of 1:1:1 to obtain a functional microorganism fermentation solution;
(2) Sterilizing bran, diatomite, biomass charcoal and fish bone powder, mixing according to a mass ratio of 8:3:3:1, adding sodium alginate and glucose, stirring and mixing uniformly, and granulating in a granulator to obtain solid particles with a particle size of 2-4 mm;
(3) And (3) adding the solid particles obtained in the step (2) into the functional microorganism fermentation liquor for adsorption, and naturally drying to obtain a final product.
The activating culture method in the step (1) comprises the following steps: and (3) thawing Trichoderma harzianum, azotobacter chroococcus and Pseudomonas fragi, respectively inoculating the Trichoderma harzianum, the azotobacter chroococcus and the Pseudomonas fragi into a PDA culture medium, culturing at 25-28 ℃, and beating bacterial colonies in a basic PDA culture medium into bacterial cakes by using a puncher of 6mm when hyphae grow to two thirds of a flat plate, so as to finish the activation culture.
The formula and the preparation method of the PDA culture medium are that 200 g of potato is boiled in water for at least 30 minutes, the filtrate is obtained after filtration, 20 g of glucose and 15-20 g of agar are added, water is added to 1000 ml, the pH value is natural, and sterilization treatment is carried out before use.
The mixed fermentation liquid culture medium comprises the following components: 5-7g glucose, 10-15g yeast powder, 0.1-0.15gKH 2PO4、0.1-0.15gMgSO4 g sodium chloride, 3-5g distilled water 1000mL, and sterilizing with steam at 121deg.C for 20min to obtain natural pH.
The method for fermentation culture in the mixed fermentation liquid culture medium comprises the following steps: culturing at 25-28deg.C, and shake culturing at 200 r/min.
Preferably, in the granulating step (2), maltose or starch is added as a binder to promote granulation.
The application of a microbial soil activation microbial agent is used for repairing and activating heavy metal contaminated soil.
More preferably, the microbial inoculum is used for repairing activated arsenic element contaminated soil.
The raw materials used in the invention are all commercially available.
The dosage of the soil microorganism activating microbial inoculum is 5-10 kg/mu. The application method comprises mixing soil restoration agent and soil to be restored uniformly, spraying water until soil humidity is 60-70%, and maintaining for 5-10 days.
Advantageous effects
(1) The trichoderma harzianum strain obtained by screening has extracellular adsorption and metal complexation effects, can realize reduction and excretion of metal ions by regulating the expression of reductase and related genes, can be combined with heavy metal ions through functional groups and proteins on the surface, and can reduce the heavy metal ions to a low valence state or a steady state through a biological reduction process, so that the heavy metal ions are fixed and removed, and particularly the trichoderma harzianum strain has higher arsenic ion removal efficiency, has the capabilities of phosphate dissolving, nitrogen fixing and IAA production, and has the effects of regulating soil and promoting crop growth;
(2) The azotobacter chroococcus not only has good nitrogen fixation capacity and adjusts the soil nutrition structure, but also can generate intracellular adsorption effect to combine heavy metal elements with complexin and the like to form thermostable proteins, so that the toxicity of the heavy metal elements is weakened or eliminated in the process; the pseudomonas fragi not only has metal complexation, but also can secrete organic acid, biosurfactant and other components, so that the activities of trichoderma harzianum and azotobacter chroococcus are improved, and the efficacy of the trichoderma harzianum and azotobacter chroococcus is promoted to be exerted;
(3) After the trichoderma harzianum and the azotobacter chroococcus and the pseudomonas fragi are combined in proportion, the interaction is obvious, and the heavy metal removal efficiency can be improved. Simultaneously, the growth is promoted in a synergic way; the three microorganisms can also regulate the content of nutrient substances in the soil, particularly regulate the activities of various enzymes in the soil, restore the ecological activity of the soil and have remarkable social and economic benefits.
Drawings
FIG. 1 is a graph showing soil urease activity of example 1 and comparative examples 1-9 of the present invention;
FIG. 2 is a graph showing the activities of soil sucrase of example 1 and comparative examples 1 to 9 of the present invention;
FIG. 3 is a graph showing the soil protease activity of example 1 and comparative examples 1 to 9 of the present invention;
FIG. 4 is a graph showing soil catalase activities of example 1 and comparative examples 1 to 9 of the present invention;
FIG. 5 is a graph showing the effect of phosphorus solubilizing ability of Trichoderma harzianum, azotobacter chroococcus and Pseudomonas fragi;
FIG. 6 is a graph showing the growth conditions of Trichoderma harzianum, azotobacter chroococcus and Pseudomonas fragi on an Ammonia-fixing culture medium of Abbe's disease, wherein A is a graph showing the growth conditions of Trichoderma harzianum, B is a graph showing the growth conditions of Pseudomonas fragi, and C is a graph showing the growth conditions of azotobacter chroococcus;
FIG. 7 is a graph showing the effect of Trichoderma harzianum, azotobacter chroococcus, and Pseudomonas fragi on producing siderophores in accordance with the present invention;
FIG. 8 is a graph showing the effect of Trichoderma harzianum, azotobacter chroococcus, and Pseudomonas fragi on the ability of Trichoderma harzianum to produce indoleacetic acid.
Detailed Description
The technical scheme of the present invention is further described below with reference to specific examples, but is not limited thereto.
Example 1
A microbial soil activation microbial agent comprises functional microbial fermentation liquor, a microbial activation agent and a microbial carrier, wherein the mass ratio of the functional microbial fermentation liquor to the microbial activation agent to the microbial carrier is 1:0.5:5.
The microbial activator is obtained by mixing sodium alginate and glucose according to a mass ratio of 1:0.5.
The microbial carrier is obtained by mixing bran, diatomite, biomass charcoal and fish bone powder according to a mass ratio of 8:3:3:1.
The functional microbial fermentation broth comprises trichoderma harzianum (Trichoderma harzianum), azotobacter chroococcus (Azotobacter chroococcum) and pseudomonas fragi (Pseudomonas plecoglossicida).
Preferably, the trichoderma harzianum (Trichoderma harzianum) has a preservation number of CGMCC No.40702, is preserved in the China general microbiological culture Collection center (China Committee for culture Collection of microorganisms), has a preservation date of 2023, 6 months and 16 days, and has a preservation address of beichen Xiyang No. 1, 3 of Beijing.
The trichoderma harzianum of the embodiment is separated from iron tailings in the mountain county of Jining, shandong province, and the content of various heavy metals including arsenic and lead in the sampling place is out of standard.
The separation method comprises the following steps: taking 5g of soil sample, placing the soil sample in a sterilized PDA culture medium, adding filtered and sterilized sodium arsenite solution, enabling the concentration of As 3+ to be 50mg/L, transferring the culture medium to a culture medium with the concentration of As 3+ of 100mg/L in an inoculum size of 2% after enrichment culture for 24 hours, culturing the culture medium under the same condition, gradually increasing the concentration of As 3+, carrying out dilution coating after transferring for 5 times, and separating and preserving the strain with the best growth vigor, namely the target strain trichoderma harzianum.
Morphological characteristics of the strain: colonies were grown on PDA plates at 25-28℃for 4 days for 8cm expansion, dark grey green; the mycelium had a septum and branches. Conidiophores form conifer-type branched outlines, and conidiophores are nearly spherical, oval, green and smooth in wall.
The nitrogen fixing bacteria have a preservation number of CGMCC No.1.5803, are purchased from the China general microbiological culture Collection center, and have an original preservation date of 6 months and 27 days in 2005; the Pseudomonas fragi has a preservation number of CGMCC No.1.16111, is purchased from the China general microbiological culture Collection center (China Committee for culture Collection of microorganisms), and has an original preservation date of 2017, 3 and 29. The azotobacter chroococcus and the pseudomonas fragi can be purchased through a collection center strain catalog, and additional collection is not needed.
The preparation method of the microbial soil activation microbial agent comprises the following preparation steps:
(1) Thawing Trichoderma harzianum, azotobacter chroococcus and Pseudomonas fragi, and respectively inoculating the Trichoderma harzianum, the azotobacter chroococcus and the Pseudomonas fragi in a PDA culture medium for activating culture; after activation culture, picking bacterial cakes of each bacterial strain, respectively adding the bacterial cakes into a mixed fermentation liquid culture medium according to an inoculation amount of 1.5% for fermentation culture, stopping fermentation when the effective viable count in each mixed fermentation liquid culture medium reaches 4X 10 8 CFU/mL to obtain three fermentation bacterial solutions, and mixing the three fermentation bacterial solutions according to a volume ratio of 1:1:1 to obtain a functional microorganism fermentation solution;
(2) Sterilizing bran, diatomite, biomass charcoal and fish bone powder, mixing according to a mass ratio of 8:3:3:1, adding sodium alginate and glucose, stirring and mixing uniformly, and granulating in a granulator to obtain solid particles with a particle size of 2-4 mm;
(3) And (3) adding the solid particles obtained in the step (2) into the functional microorganism fermentation liquor for adsorption, and naturally drying to obtain a final product.
The activating culture method in the step (1) comprises the following steps: and (3) thawing Trichoderma harzianum, azotobacter chroococcus and Pseudomonas fragi, respectively inoculating the Trichoderma harzianum, the azotobacter chroococcus and the Pseudomonas fragi into a PDA culture medium, culturing at 25-28 ℃, and beating bacterial colonies in a basic PDA culture medium into bacterial cakes by using a puncher of 6mm when hyphae grow to two thirds of a flat plate, so as to finish the activation culture.
The formula and the preparation method of the PDA culture medium are that 200 g of potato is boiled in water for at least 30 minutes, the filtrate is obtained after filtration, 20 g of glucose and 15-20 g of agar are added, water is added to 1000 ml, the pH value is natural, and sterilization treatment is carried out before use.
The mixed fermentation liquid culture medium comprises the following components: 5-7g glucose, 10-15g yeast powder, 0.1-0.15gKH 2PO4、0.1-0.15gMgSO4 g sodium chloride, 3-5g distilled water 1000mL, and sterilizing with steam at 121deg.C for 20min to obtain natural pH.
The method for fermentation culture in the mixed fermentation liquid culture medium comprises the following steps: culturing at 25-28deg.C, and shake culturing at 200 r/min.
And (3) during granulation in the step (2), adding starch as a binder to promote granulation.
The application of a microbial soil activation microbial agent is used for repairing and activating heavy metal contaminated soil.
The microbial inoculum is used for repairing activated arsenic element polluted soil.
The raw materials used are commercially available.
The dosage of the soil microorganism activating bacterial agent is 5-10 kg/mu. The application method comprises mixing soil restoration agent and soil to be restored uniformly, spraying water until soil humidity is 60-70%, and maintaining for 5-10 days.
Example 2
A microbial soil activation microbial agent comprises functional microbial fermentation liquor, a microbial activation agent and a microbial carrier, wherein the mass ratio of the functional microbial fermentation liquor to the microbial activation agent to the microbial carrier is 1:0.5:5.
The microbial activator is obtained by mixing sodium alginate and glucose according to a mass ratio of 1:1.5.
The microbial carrier is obtained by mixing bran, diatomite, biomass charcoal and fish bone powder according to a mass ratio of 5:5:4:2.
The functional microbial fermentation broth comprises trichoderma harzianum (Trichoderma harzianum), azotobacter chroococcus (Azotobacter chroococcum) and pseudomonas fragi (Pseudomonas plecoglossicida).
The trichoderma harzianum (Trichoderma harzianum) has a preservation number of CGMCC No.40702, is preserved in the China general microbiological culture Collection center (China Committee for culture Collection of microorganisms), has a preservation date of 2023, 6 and 16 days, and has a preservation address of beichen Xiya No. 1, 3 in the Korean region of Beijing.
Trichoderma harzianum is isolated from iron tailings at a place in Weishan county, jining, shandong province, and the isolation method is the same as in example 1.
The azotobacter chroococcus and the Pseudomonas fragi are selected as in example 1.
A preparation method of a microbial soil activation microbial agent. As in example 1.
In the step (2), maltose is added as a binder for granulation.
Example 3
A microbial soil activation microbial agent comprises functional microbial fermentation liquor, a microbial activation agent and a microbial carrier, wherein the mass ratio of the functional microbial fermentation liquor to the microbial activation agent to the microbial carrier is 1:0.5:5.
The microbial activator is obtained by mixing sodium alginate and glucose according to a mass ratio of 1:1.
The microbial carrier is obtained by mixing bran, diatomite, biomass charcoal and fish bone powder according to a mass ratio of 6:4:6:3.
The functional microbial fermentation broth comprises trichoderma harzianum (Trichoderma harzianum), azotobacter chroococcus (Azotobacter chroococcum) and pseudomonas fragi (Pseudomonas plecoglossicida).
The trichoderma harzianum (Trichoderma harzianum) has a preservation number of CGMCC No.40702, is preserved in the China general microbiological culture Collection center (China Committee for culture Collection of microorganisms), has a preservation date of 2023, 6 and 16 days, and has a preservation address of beichen Xiya No. 1, 3 in the Korean region of Beijing.
Trichoderma harzianum is isolated from iron tailings at a place in Weishan county, jining, shandong province, and the isolation method is the same as in example 1.
The azotobacter chroococcus and the Pseudomonas fragi are selected as in example 1.
A preparation method of a microbial soil activation microbial agent. As in example 1.
And (3) during granulation in the step (2), adding starch as a binder to promote granulation.
Example 4
Research on growth promotion of bacterial strain
The qualitative tests of phosphate dissolving, nitrogen fixing, siderophore producing and indoleacetic acid producing are respectively carried out by exploring whether all three selected strains have growth promoting effect, and the test method is as follows:
1) Determination of phosphate-solubilizing ability
Preparing a PKO solid culture medium, diluting and coating bacterial liquid which is activated and cultured to OD 600 =0.8-1.0 to a flat culture medium, and culturing for 3d at 28-30 ℃, wherein the bacterial strain with the phosphate dissolving effect can form transparent phosphate dissolving rings around bacterial colonies.
PKO medium composition was: glucose 10g、(NH4)SO4 0.5g、NaCl 0.3g、KCl 0.3g、MgSO4·7H2O 0.3g、FeSO4·7H2O 0.03g、MnSO4·4H2O 0.03g、Ca3(PO4)2 5g、 agar 20g, distilled water 1000mL, pH7.0-7.2.
2) Nitrogen fixation Capacity determination
The activated bacterial liquid is diluted and coated on an Ababetes ammonia fixation culture medium in a gradient way, the culture is carried out for 3 days at the constant temperature of 28-30 ℃, the formation of aseptic colonies on a flat plate is observed, and single bacterial colonies are continuously transferred for 3 times, so that the bacterial strain capable of stably growing has the nitrogen fixation capability.
The composition of the Ababetes ammonia fixation culture medium is as follows: k 2HPO4 0.2g、NaCl 0.2 g,CaCO3 2.0.0 g, mannitol 10g, caSO 4 0.1g、MgSO4 0.2.2 g, agar 1g, distilled water 1000 mL, pH6.9.
3) Determination of iron production carrier capacity
The separated and purified strain is inoculated on a CAS detection plate by a partition point inoculation method, cultured for 3d at 20-30 ℃, and the colony is observed, if obvious orange-yellow halo exists around the colony, the strain is proved to have the capacity of producing the iron carrier.
The CAS detection plate consists of: 20% sucrose solution, 10mL10% casein hydrolysate 30 mL, 1mmol/LCaCl 21mL、1 mmol/LMgSO4 mL, 0.1mol/L buffer 5mL, CAS dye 5mL, agar 18g, distilled water 1000mL.
4) Determination of indoleacetic acid (IAA) production Capacity
The strain to be tested is inoculated in King' B culture medium of 50 mL, after constant temperature culture for 3d, bacterial suspension is centrifuged for 6min at 8000r/min, 500 mu L of supernatant and 500 mu L of Salkowski colorimetric solution are respectively taken, added into a centrifuge tube of 1.5mL, and the mixture is kept stand in the dark for 30 min, and color development is waited. In addition, the bacterial solutions were replaced with equal amounts of non-inoculated blank medium and 50 mg/L IAA, respectively, as negative and positive controls. The color change can be used for preliminarily judging whether the strain has the capability of producing IAA, and the darker the color is, the more the IAA secretion amount of the strain is represented.
King' B medium consisted of: 20.0g of peptone, 1.5g of K 2HPO41.5g、MgSO4·7H2 O, 1000mL of distilled water and pH7.2.
TABLE 1 qualitative determination of the growth-promoting Capacity of strains
Note that: "+" is active and "-" is inactive.
Comparative examples 1 to 9
The composition of the functional microbial fermentation broth, i.e., the volume ratio of trichoderma harzianum, azotobacter chroococcus and pseudomonas meracillus, was changed, and a comparative example was set. The rest of the raw materials and the process are the same as in example 1. The preparation method comprises the following steps:
Thawing Trichoderma harzianum, azotobacter chroococcus and Pseudomonas fragi, and respectively inoculating the Trichoderma harzianum, the azotobacter chroococcus and the Pseudomonas fragi in a PDA culture medium for activating culture; after activation culture, bacterial cakes of all strains are selected and respectively added into a mixed fermentation liquid culture medium for fermentation culture according to the inoculation amount of 1.5%, when the effective viable count in each mixed fermentation liquid culture medium reaches 4X 10 8 CFU/mL, fermentation is stopped to obtain three fermentation bacterial solutions, and then the three fermentation bacterial solutions are mixed according to different volume ratios to obtain the functional microbial fermentation liquid.
The volume ratios of trichoderma harzianum, azotobacter chroococcus and pseudomonas fragi in the comparative examples are shown in table 2:
TABLE 2 comparative example seed mass ratio
Determination of heavy metal ion removal ability of Strain
The functional microorganism fermentation broths obtained in example 1 and comparative examples 1 to 9 of the present invention were inoculated in an amount of 1% to 100mL of a liquid medium (NB) containing Pb 2+ at a concentration of 150mg/L, as 5+ and 200mg/L at a concentration of 100mg/L, cd 2+, cultured at 25-28℃under shaking at 180r/min for 48 hours, centrifuged at 5000 r/min for 10 min, and the supernatant was collected for ICP-MS to determine the concentrations of Pb 2+、As5+、Cd2+ and Pb in the supernatant. And calculating the heavy metal ion removal rate by using a formula. The formula is:
Wherein, C 0 is the concentration of the original metal ions in the culture medium, and C e is the concentration of the residual metal ions after treatment. The units are mg/L. The removal results are shown in table 3:
Table 3 results of the removal experiments
Soil remediation experiment:
test soil: selecting soil polluted by heavy metals near certain electroplating plants in east region of Linyi city
The test method comprises the following steps: taking polluted soil, wherein the blank group treatment directly loads the tested soil into a flowerpot with the length of about 20cm multiplied by 20 cm; the other experimental groups were added with the microbial activators obtained in examples 1-3 and comparative examples 1-9 according to 5 kg/mu (about 1.4% of soil weight), mixed uniformly, charged into flowerpots, each pot filled with 1kg of soil, 3 parallel samples were set for each group of experiments, and the results were averaged. Deionized water is added by a weighing method to maintain the water content of the soil to about 60 percent. After one week of maintenance, collecting a soil sample, removing impurities, naturally air-drying, grinding, and respectively sieving with 10 mesh and 100 mesh sieve for storage.
The analytical test method comprises the following steps:
Soil microorganisms were determined using a dilution plate separation method. Soil urease activity was determined using indophenol colorimetry. The activity of the sucrase is determined by a colorimetric method of 3, 5-dinitrosalicylic acid. The catalase activity was determined by potassium permanganate titration. Protease activity was measured using a modified ninhydrin colorimetric method.
The soil sample is naturally dried and sieved, and then the pH value of the soil is directly read out by an acidimeter and a conductivity meter according to a soil-water ratio of 1:5 leaching method; the quick-acting nitrogen content is determined by adopting an alkaline hydrolysis diffusion method, the quick-acting phosphorus content and the quick-acting potassium content are determined by adopting a combined leaching-colorimetric method, and the organic matter content is determined by adopting a potassium dichromate capacity method. The heavy metal ion content of the rhizosphere soil was determined with an atomic absorption spectrophotometer (PERKINELMER SIMMA6000,6000, norwalk, USA).
Table 4 soil index test results
TABLE 5 soil index test results
TABLE 6 soil index test results
According to the performance data, the soil repaired by the embodiment of the invention can effectively control the content level of heavy metal elements in the soil under the activation of three functional strains, and meanwhile, the effective activity of microorganisms, the content of soil nutrients, biomass and enzyme activity level are obviously improved. The synergistic balance between strains is broken by changing the composition and the dosage of the strain in comparative examples 1-9, and the action generated in the microbial activity process is weakened, so that the soil restoration effect is weakened.
It should be noted that the above-mentioned embodiments are merely some, but not all embodiments of the preferred mode of carrying out the invention. It is evident that all other embodiments obtained by a person skilled in the art without making any inventive effort, based on the above-described embodiments of the invention, shall fall within the scope of protection of the invention.
Claims (9)
1. The microbial soil activation microbial agent is characterized by comprising functional microbial fermentation liquor, a microbial activator and a microbial carrier, wherein the mass ratio of the functional microbial fermentation liquor to the microbial activator to the microbial carrier is 1:0.5:5.
2. The microbial soil activation microbial agent according to claim 1, wherein the microbial agent is obtained by mixing sodium alginate and glucose according to a mass ratio of 1 (0.5-1.5).
3. The microbial soil activation microbial agent according to claim 1, wherein the microbial carrier is prepared by mixing (5-8): 3-5): 3-6): 1-3 by mass ratio of bran, diatomite, biomass charcoal and fish bone powder.
4. The microbial soil activation microbial agent of claim 1, wherein the functional microbial fermentation broth comprises trichoderma harzianum (Trichoderma harzianum), azotobacter chroococcus (Azotobacter chroococcum), and pseudomonas fragi (Pseudomonas plecoglossicida).
5. The microbial soil activation microbial agent according to claim 4, wherein the trichoderma harzianum has a preservation number of CGMCC No.40702, and is preserved in the China general microbiological culture collection center (cmm); the preservation number of the azotobacter chroococcus is CGMCC No.1.5803, and the azotobacter chroococcus is purchased from the China general microbiological culture Collection center; the Pseudomonas fragi has a preservation number of CGMCC No.1.16111 and is purchased from the China general microbiological culture Collection center (China Committee for culture Collection of microorganisms).
6. A method for preparing the microbial soil activation microbial agent as claimed in any one of claims 1 to 5, comprising the following preparation steps:
(1) Thawing Trichoderma harzianum, azotobacter chroococcus and Pseudomonas fragi, and respectively inoculating the Trichoderma harzianum, the azotobacter chroococcus and the Pseudomonas fragi in a PDA culture medium for activating culture; after activation culture, picking bacterial cakes of each bacterial strain, respectively adding the bacterial cakes into a mixed fermentation liquid culture medium according to the inoculation amount of 1.5% for fermentation culture, stopping fermentation when the effective viable count in the mixed fermentation liquid culture medium reaches 4X 10 8 CFU/mL to obtain three fermentation bacterial solutions, and mixing the three fermentation bacterial solutions according to the volume ratio of 1:1:1 to obtain functional microbial fermentation liquor;
(2) Sterilizing bran, diatomite, biomass charcoal and fish bone powder, mixing according to a mass ratio of 8:3:3:1, adding sodium alginate and glucose, stirring and mixing uniformly, and granulating in a granulator to obtain solid particles with a particle size of 2-4 mm;
(3) And (3) adding the solid particles obtained in the step (2) into the functional microorganism fermentation liquor for adsorption, and naturally drying to obtain a final product.
7. The method for producing a microbial soil-activating microbial agent according to claim 6, wherein maltose or starch is added as a binder during the granulation in the step (2) to promote the granulation.
8. Use of a microbial soil activation microbial agent as claimed in any one of claims 1 to 5, wherein the microbial agent is for repairing activated heavy metal contaminated soil.
9. The use of a microbial soil activation microbial agent as claimed in claim 8, wherein the microbial agent is used for remediation of activated arsenic element contaminated soil.
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Citations (14)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20040261578A1 (en) * | 2003-04-04 | 2004-12-30 | Harman Gary E | Stable self-organizing plant-organism systems for remediating polluted soils and waters |
| US20050126990A1 (en) * | 2001-11-23 | 2005-06-16 | Lynch James M. | Bioremediation |
| US20120015806A1 (en) * | 2009-03-25 | 2012-01-19 | Sitaram Prasad Paikray | Novel formulation of microbial consortium based bioinoculant for wide spread use in agriculture practices |
| CN103467154A (en) * | 2013-08-14 | 2013-12-25 | 中国人民解放军济南军区72465部队 | Safety disposal and resource utilization method of lead-containing energetic material |
| CN106118683A (en) * | 2016-06-30 | 2016-11-16 | 山东爱福地生物科技有限公司 | A kind of heavy-metal contaminated soil renovation agent |
| KR20170043768A (en) * | 2015-10-14 | 2017-04-24 | 라광수 | Method for manufacturing manure by microorganism using organic waste and the manure manufactured thereof |
| CN107522541A (en) * | 2017-08-24 | 2017-12-29 | 宁夏顺宝现代农业股份有限公司 | Biological organic fertilizer produced using chicken manure and furfural dregs and preparation method thereof |
| WO2018011007A1 (en) * | 2016-07-13 | 2018-01-18 | Empa Eidgenössische Materialprüfungs- Und Forschungsanstalt | Superior antagonistic trichoderma species for preserving plant material and wood products |
| CN113149785A (en) * | 2021-03-25 | 2021-07-23 | 上海又然生态科技有限公司 | Bio-organic fertilizer capable of effectively passivating soil heavy metals and preparation method thereof |
| CN113817626A (en) * | 2020-12-07 | 2021-12-21 | 内蒙古百奥科技有限公司 | Pseudomonas brassicae Y1, culture method and application thereof, and soil conditioner |
| CN114621890A (en) * | 2021-03-24 | 2022-06-14 | 施可丰化工股份有限公司 | Compound microbial agent for adsorbing and removing heavy metal elements in soil and preparation method thereof |
| WO2022203951A1 (en) * | 2021-03-20 | 2022-09-29 | Locus Agriculture Ip Company, Llc | Holistic and environmentally-friendly systems for crop, soil, water and livestock management |
| CN115572187A (en) * | 2022-08-29 | 2023-01-06 | 中昇南阳生物科技有限公司 | Microbial compound fertilizer and method for repairing heavy metal contaminated soil by using same |
| CN117089355A (en) * | 2023-08-24 | 2023-11-21 | 临沂市农业科学院 | Soil heavy metal pollution restoration agent and preparation method thereof |
-
2024
- 2024-03-14 CN CN202410288031.5A patent/CN117903813B/en active Active
Patent Citations (14)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20050126990A1 (en) * | 2001-11-23 | 2005-06-16 | Lynch James M. | Bioremediation |
| US20040261578A1 (en) * | 2003-04-04 | 2004-12-30 | Harman Gary E | Stable self-organizing plant-organism systems for remediating polluted soils and waters |
| US20120015806A1 (en) * | 2009-03-25 | 2012-01-19 | Sitaram Prasad Paikray | Novel formulation of microbial consortium based bioinoculant for wide spread use in agriculture practices |
| CN103467154A (en) * | 2013-08-14 | 2013-12-25 | 中国人民解放军济南军区72465部队 | Safety disposal and resource utilization method of lead-containing energetic material |
| KR20170043768A (en) * | 2015-10-14 | 2017-04-24 | 라광수 | Method for manufacturing manure by microorganism using organic waste and the manure manufactured thereof |
| CN106118683A (en) * | 2016-06-30 | 2016-11-16 | 山东爱福地生物科技有限公司 | A kind of heavy-metal contaminated soil renovation agent |
| WO2018011007A1 (en) * | 2016-07-13 | 2018-01-18 | Empa Eidgenössische Materialprüfungs- Und Forschungsanstalt | Superior antagonistic trichoderma species for preserving plant material and wood products |
| CN107522541A (en) * | 2017-08-24 | 2017-12-29 | 宁夏顺宝现代农业股份有限公司 | Biological organic fertilizer produced using chicken manure and furfural dregs and preparation method thereof |
| CN113817626A (en) * | 2020-12-07 | 2021-12-21 | 内蒙古百奥科技有限公司 | Pseudomonas brassicae Y1, culture method and application thereof, and soil conditioner |
| WO2022203951A1 (en) * | 2021-03-20 | 2022-09-29 | Locus Agriculture Ip Company, Llc | Holistic and environmentally-friendly systems for crop, soil, water and livestock management |
| CN114621890A (en) * | 2021-03-24 | 2022-06-14 | 施可丰化工股份有限公司 | Compound microbial agent for adsorbing and removing heavy metal elements in soil and preparation method thereof |
| CN113149785A (en) * | 2021-03-25 | 2021-07-23 | 上海又然生态科技有限公司 | Bio-organic fertilizer capable of effectively passivating soil heavy metals and preparation method thereof |
| CN115572187A (en) * | 2022-08-29 | 2023-01-06 | 中昇南阳生物科技有限公司 | Microbial compound fertilizer and method for repairing heavy metal contaminated soil by using same |
| CN117089355A (en) * | 2023-08-24 | 2023-11-21 | 临沂市农业科学院 | Soil heavy metal pollution restoration agent and preparation method thereof |
Non-Patent Citations (4)
| Title |
|---|
| SARIKA DONGA等: "Nutrients Uptake by Groundnut (Arachis hypogaea L.) and Soil Fertility under the Influence of Enriched Vermicompost", BIOLOGICAL FORUM – AN INTERNATIONAL JOURNAL, vol. 13, no. 2, 31 December 2021 (2021-12-31), pages 149 - 152 * |
| 卜和申等: "哈茨木霉的耐镉性及其对狗牙根耐镉能力的影响", 草地学报, no. 01, 15 January 2020 (2020-01-15), pages 67 - 74 * |
| 袁扬等: "木霉菌在农业中的应用研究进展", 江苏农业科学, no. 03, 8 March 2018 (2018-03-08), pages 18 - 22 * |
| 车永梅等: "AM真菌、木霉和PGPR组合的促生效应研究", 青岛农业大学学报(自然科学版), no. 02, 15 June 2019 (2019-06-15), pages 20 - 27 * |
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