CN112500865B - Alkaline soil remediation microbial inoculum and preparation method thereof - Google Patents
Alkaline soil remediation microbial inoculum and preparation method thereof Download PDFInfo
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Abstract
The application relates to the field of microbial agents, and particularly discloses a saline-alkali soil remediation microbial inoculum and a preparation method thereof, wherein the saline-alkali soil remediation microbial inoculum comprises the following substances in parts by weight: 25-30 parts of a microbial agent carrier, 10-15 parts of a composite microbial agent and 25-30 parts of biochemical organic acid, wherein the composite microbial agent comprises Bacillus amyloliquefaciens, Paenibacillus mucilaginosus, Bacillus megatherium and Trichoderma harzianum which are mixed according to the mass ratio of 1-2: 5; the microbial inoculum carrier is attapulgite, and the biochemical organic acid comprises biochemical fulvic acid and fulvic acid which are mixed in equal mass. The composition of microbial inoculum has been optimized in this application, has improved the defect of single bacterial microbial inoculum, adopts the attapulgite simultaneously as the matrix, and the supplementary organic acid that uses improves the material as supplementary, improves saline and alkaline soil remediation rate to prolong its improvement life-span, finally effectively improve saline and alkaline soil.
Description
Technical Field
The application relates to the field of microbial agents, in particular to an alkaline soil remediation microbial agent and a preparation method thereof.
Background
The alkaline soil is a soil which is formed in a salinization process under the comprehensive action of various factors such as artificial activities, natural environment and the like, contains a large amount of soluble salts, has a salinization layer or an alkalization layer and can not enable plants to normally grow. In the soil, exchangeable sodium continuously enters a soil colloid process, particularly an organic-mineral absorption complex, which is also called as a sodium transformation process, when calcium ions of the soil colloid are completely replaced by sodium ions, saturated sodium ions are desorbed from the colloid into a soil solution, so that alkaline soil is high in salt content and alkalinity, soil humus is easily leached, a soil structure is damaged, the soil structure is sticky when wet and hardened when dry, ventilation and water permeation are poor, and plant wilting, poisoning and root rot death can be caused after plants are planted.
Soil microorganisms are one of the most active factors of soil fertility, and the application of the microbial remediation microbial inoculum can effectively remediate soil and improve soil fertility. Some microorganisms can generate organic acid in the metabolic process, provide slow-acting nitrogen, phosphorus and potassium for soil, improve the physicochemical property of alkaline soil, effectively reduce the pH and the salt content of saline-alkali soil, improve the organic matter and nutrient content of the alkaline soil, and improve the survival rate and the yield of crops.
In view of the above-mentioned related technologies, the inventors believe that in the existing scheme of improving alkaline soil by using microbial agents, the pH and salt content of soil are improved only by metabolizing organic acids after the propagation of microorganisms, the effect is slow, the durability is not high, the improvement effect is slow, and meanwhile, part of microbial agents are slowly inactivated in the long-term improvement process, so that the effect of continuously repairing soil is reduced.
Disclosure of Invention
In order to overcome the defects that the alkaline soil remediation microbial inoculum is low in remediation rate and low in durability in the alkaline soil remediation process, the application provides the alkaline soil remediation microbial inoculum, and the following technical scheme is adopted:
an alkaline soil remediation microbial inoculum comprises the following substances in parts by weight: 25-30 parts of a microbial inoculum carrier, 10-15 parts of a composite microbial inoculum and 25-30 parts of biochemical organic acid; the composite microbial agent comprises Bacillus amyloliquefaciens, Bacillus mucilaginosus, Bacillus megatherium and Trichoderma harzianum which are mixed according to the mass ratio of 1-2: 5; the microbial inoculum carrier is attapulgite, and the biochemical organic acid comprises biochemical fulvic acid and fulvic acid which are mixed in equal mass.
By adopting the technical scheme, the composition of the microbial agent is optimized, and the alkaline soil is improved by adopting a multi-strain combined microbial agent scheme, wherein the bacillus can effectively activate silicate minerals, dissolve phosphorus, dissolve potassium and fix nitrogen, so that the soil texture and the micro-ecological environment are improved, trichoderma harzianum is fixedly planted on plant cells, and the protection is provided when the trichoderma harzianum is infected by soil-borne pathogenic bacteria, so that the soil-borne diseases can be effectively prevented and treated, the two types of microorganisms have synergistic action, and the defect of the single-strain microbial agent is improved; on this basis, this application adopts the attapulgite as the matrix, and the supplementary organic acid that uses improves the material as the supplementary, and the purpose lies in through selecting for use suitable load matrix, can make microbial inoculum initial stage reproductive efficiency effectively improve, and the preliminary neutralization of rethread organic acid provides good reproductive environment, and the buffering that constitutes with the various hydroxyl and the phenolic hydroxyl that contain inside simultaneously is right, reduces soil pH value, can effectual buffering and alleviate the salt damage and the alkali damage of soil, improves alkaline soil remediation speed to prolong its improvement life, finally effectively improve alkaline soil.
Further, the microbial inoculum carrier is calcined and modified attapulgite, the exchangeable calcium ion amount of the calcined and modified attapulgite is 7.5-12.5 mmol/100g, and the exchangeable magnesium ion amount is 2.5-7.5 mmol/100 g.
Through adopting above-mentioned technical scheme, because this application is modified through calcining the processing to the attapulgite carrier, because high temperature calcination handles, has got rid of its inside loaded zeolite water, adsorbed water and crystal water, the needle bar form fibre group of unordered structure becomes loose inflation in the messenger attapulgite structure to the pore volume and the specific surface area of attapulgite have been improved, through calcining the structure after the modification, improve soil colloid performance, and further improve its load effect who carries the microbial inoculum, thereby improve the life-span of microbial inoculum material, finally effectively improve alkaline soil.
Further, the microbial inoculum carrier is calcined modified attapulgite coated by an adjusting modified coating solution, and the modified coating solution is humic acid ammonium-acrylamide graft copolymer.
By adopting the technical scheme, because the calcined and modified attapulgite is modified, the attapulgite is calcined at high temperature to remove zeolite water, adsorbed water and crystal water, so that the structure of the attapulgite is loosened and expanded, so that the internal pore structure thereof is collapsed and destroyed under high-temperature treatment, so that the graft coating solution is modified, by the coating modification scheme, the coating liquid is used for protecting the pores in the attapulgite, so that the graft copolymer permeates into the pore structure in the calcined attapulgite, so that after drying and curing, a coating layer is formed inside the pores after the coated gel is cured, the coating layer effectively covers and supports the inner walls of the pores, thereby improving the stability of the calcined attapulgite, improving the loading effect of the loaded microbial inoculum and the service life of the microbial inoculum material, and finally effectively improving the alkaline soil.
Further, the humic acid ammonium-acrylamide graft copolymer is prepared by the following method: (1) stirring and mixing humic acid, ammonia water and deionized water according to a mass ratio of 1: 3-5: 10-15, collecting mixed liquor, rotating and evaporating to dryness, collecting dry particles, and mixing the dry particles according to a mass ratio of 9-10: 3-5: 1 stirring and mixing deionized water, acrylamide and dry particles; (2) and in a nitrogen atmosphere, taking an ammonium persulfate solution as an initiator, carrying out heat preservation reaction at 75-80 ℃ in the nitrogen atmosphere, standing and cooling to room temperature to obtain the coating modification liquid.
By adopting the technical scheme, because the scheme of grafting polyacrylamide with ammonium humate is adopted to prepare the coating modified solution, alkaline soil is applied by the ammonium humate to reduce the alkalinity of the soil, organic anions and phosphorus compete for adsorption sites to be combined by chelation, the fixation of the soil to phosphorus and potassium is reduced, phosphorus and potassium elements in insoluble state and fixed state are decomposed, on the basis, the structure of soil aggregates is effectively maintained by the polyacrylamide to form new aggregates, the formation of crusts is inhibited by the flocculation effect formed by interaction with water, the infiltration capacity of the soil can be increased, and as the coating modified solution has a spatial network structure and can be mutually associated with molecular groups and water molecules, the absorption and water retention performances can effectively reduce the soil erosion and simultaneously form a good microbial propagation environment, so the improvement efficiency of the microbial inoculum in the initial stage is further improved, thereby prolonging the service life of the microbial inoculum material and finally effectively improving the alkaline soil.
Furthermore, the total effective viable count of the alkaline soil remediation microbial inoculum is not less than 15.0 hundred million cfu/g, and the mixed bacteria rate of the alkaline soil remediation microbial inoculum is not more than 20%.
By adopting the technical scheme, the number of effective viable bacteria of the prepared soil remediation microbial inoculum is increased, so that beneficial bacteria in soil are greatly increased, the air permeability of the soil after remediation can be obviously improved in the primary improvement period, the soil hardening problem is effectively solved, and the micro-ecological environment of the soil is improved.
Further, the microbial inoculum carrier is prepared by adopting the following method: (1) calcining attapulgite at 250-300 ℃, standing and cooling to obtain a modified attapulgite matrix; (2) and (3) stirring and mixing the coating modification solution and the modified attapulgite matrix, carrying out heat preservation and pressurization stirring treatment at 55-60 ℃ under 5-8 MPa, filtering, taking a filter cake, drying, collecting dry particles, and grinding to obtain the microbial inoculum carrier.
By adopting the technical scheme, the modified attapulgite is coated and calcined by the coating modification liquid to prepare the microbial inoculum carrier, the coated attapulgite has good structural performance, so that the effective load of the microbial inoculum is in the pores inside the attapulgite, and meanwhile, the polyacrylamide in the coating modification liquid effectively maintains the structure of soil aggregates, and the molecular groups and water molecules can be mutually associated to provide a good propagation environment for microorganisms in the microbial inoculum, so that the service life of the microbial inoculum material is prolonged, and finally, the alkaline soil is effectively improved.
In a second aspect, the present application provides a method for preparing an alkaline soil remediation microbial inoculum, wherein the preparation method of the alkaline soil remediation microbial inoculum comprises the following steps: s1, preparing mixed strains: respectively activating and culturing bacillus amyloliquefaciens, bacillus mucilaginosus, bacillus megaterium and trichoderma harzianum spore powder according to the formula, and collecting thalli to obtain mixed strains; s2, preparation of a culture medium carrier: respectively taking diatomite, turf and bran, and carrying out jet milling and sieving to obtain a culture medium carrier; s3, preparing a complex microbial inoculum: stirring and mixing the mixed strain and a culture medium carrier, keeping the temperature at 35-40 ℃, drying, and crushing to obtain a composite microbial inoculum; s4, preparing an alkaline soil remediation microbial inoculum: and then stirring and mixing the composite microbial inoculum, the biochemical organic acid and a microbial inoculum carrier according to a formula, and grinding and dispersing in a mortar to prepare the alkaline soil remediation microbial inoculum.
By adopting the technical scheme, because the microorganism is loaded in a drying mode, the microbial activity in the microbial inoculum is not influenced during treatment, the water content after drying is low, the metabolic activity of the microorganism is reduced, and the preservation time of the solidified microbial inoculum is effectively prolonged.
Further, the size of the screen mesh sieved in step S2 is 500 meshes.
Through adopting above-mentioned technical scheme, this application is through the particle diameter of optimizing the culture medium carrier, and the bacterial of the culture medium base member internal load after the optimization can not lead to nutrient substance and the not good problem of reproductive performance that leads to of reproduction carrier space inadequately when using the initial stage and can form the reproduction of at utmost to can the at utmost optimize the treatment effeciency in early stage of microbial inoculum, thereby can improve alkaline soil repair rate, thereby prolong its improvement life-span, finally effectively improve alkaline soil.
In summary, the present application includes at least one of the following beneficial technical effects:
firstly, the application adopts the coating modification liquid to coat calcined and modified attapulgite to prepare a microbial inoculum carrier, and the coated attapulgite has good structural performance, so that the microbial inoculum is effectively loaded in pores inside the attapulgite.
Secondly, the composition of the microbial agent is optimized, the soil texture and the micro-ecological environment are improved through the bacillus, the soil-borne diseases are prevented and controlled by the trichoderma harzianum, and the defects of the microbial agent with a single strain are improved through the synergistic effect of the two types of microorganisms.
Thirdly, the scheme of grafting polyacrylamide with ammonium humate is adopted to prepare the coating modified solution, alkaline soil is applied through the ammonium humate firstly, the alkalinity of the soil is reduced, meanwhile, the fixation of the soil to phosphorus and potassium is reduced through the chelation effect, and on the basis, the structure of soil aggregates is effectively maintained through the polyacrylamide.
Detailed Description
The present application will be described in further detail with reference to examples.
In the examples of the present application, the following instruments and apparatuses are used, but not limited thereto:
a machine: pH meter, continuous flow analyzer.
Examples
Example 1
Culturing and collecting trichoderma harzianum inoculum: taking 400g of wood, 100g of bran and 200g of soybean meal, naturally drying and crushing, performing ultraviolet sterilization treatment, and collecting crushed particles to obtain a solid fermentation substrate; 4500mL of deionized water, 300g of starch granules and 100g of sodium nitrate are respectively weighed and placed in a stirrer, stirred and dispersed at room temperature, and fermentation nutrient solution is collected; spraying a fermentation nutrient solution onto the surface of a solid fermentation substrate, controlling the water content of the solid fermentation substrate to be 55%, inoculating trichoderma harzianum into the solid fermentation substrate in a dispersed manner according to the inoculation mass concentration of 7.5g/kg, culturing the trichoderma harzianum by using a freshness protection package bag with small holes filled, fermenting and culturing the trichoderma harzianum at 26 ℃ for 140 hours, opening the packaging bag and naturally drying the trichoderma harzianum for 24 hours after the fermentation culture is finished, stirring and mixing the air-dried fermentation product and sterile water according to the mass ratio of 1:8, filtering the mixture, collecting spore suspension, filtering the mixture for the second time and naturally drying the mixture to obtain dry spore;
culturing and collecting a bacillus agent: weighing 1500g of corn flour, 150g of composite powder formed by mixing yeast powder, soybean meal and the like in mass, 60g of sodium chloride, 2g of manganese sulfate, 20g of dipotassium hydrogen phosphate and 10L of water respectively, stirring, mixing, performing ultraviolet sterilization treatment, and collecting to obtain a liquid culture medium; respectively taking bacillus amyloliquefaciens, bacillus mucilaginosus and bacillus megaterium, respectively inoculating the bacillus amyloliquefaciens, the bacillus mucilaginosus and the bacillus megaterium into a liquid culture medium according to the inoculation volume ratio of 6%, performing heat preservation fermentation treatment at 38 ℃ for 50 hours, collecting fermentation liquor, performing centrifugal separation at 1500r/min, collecting lower-layer sediment, performing vacuum freeze drying, and mixing 20g of vacuum freeze-dried bacillus amyloliquefaciens, 20g of bacillus mucilaginosus and 20g of bacillus megaterium to obtain bacillus strains;
preparation of a culture medium carrier: respectively weighing 4500g of diatomite, 1500g of turf and 1000g of bran, performing jet milling, and sieving with a 500-mesh sieve to obtain a culture medium carrier;
preparing a complex microbial inoculum: stirring and mixing 50g of dry spore powder, 30g of bacillus strain and 150g of culture medium carrier, keeping the temperature and drying at 35 ℃, crushing, and sieving with a 200-mesh sieve to obtain a composite microbial inoculum;
preparing a coating modifying solution: stirring and mixing 100g of humic acid, 300mL of 1% ammonia water in mass fraction and 1000mL of deionized water, collecting the mixed solution, placing the mixed solution in a rotary evaporation device, magnetically stirring and mixing for 2 hours at 110 ℃, obtaining the mixed solution, placing the mixed solution in the rotary evaporation device, rotatably evaporating to dryness, and collecting dry particles; respectively weighing 4500mL of deionized water, 1000g of acrylamide and 300g of dry particles, placing the deionized water, the acrylamide and the dry particles in a reaction container, stirring and mixing, dropwise adding an ammonium persulfate solution with the mass fraction of 1% in the reaction container under the nitrogen atmosphere, controlling the dropwise adding amount to be 1% of the mass of the acrylamide, after the dropwise adding is finished, carrying out heat preservation reaction for 2 hours under the nitrogen atmosphere at the temperature of 75 ℃, standing and cooling to room temperature to obtain a coating modification solution;
preparation of a microbial inoculum carrier: taking attapulgite, placing the attapulgite in a muffle furnace, calcining at 250 ℃ for 2h, standing and cooling to room temperature to obtain a modified attapulgite matrix, taking 100mL of coating modification solution, stirring and mixing with 1500g of the modified attapulgite matrix, stirring for 2h at 55 ℃ and under the pressure of 5MPa, filtering after the pressure stirring is finished, taking a filter cake, placing the filter cake at 100 ℃ for drying for 2h, collecting dry particles, grinding, and sieving with a 200-mesh sieve to obtain a microbial inoculum carrier;
preparing an alkaline soil remediation microbial inoculum: stirring and mixing 10g of the composite microbial inoculum, 20g of the mixed biochemical fulvic acid and ulmic acid with equal mass and 20g of a microbial inoculum carrier, and grinding and dispersing in a mortar to prepare the alkaline soil remediation microbial inoculum.
Example 2
Culturing and collecting trichoderma harzianum inoculum: naturally drying 450g of wood, 120g of bran and 250g of soybean meal, crushing, performing ultraviolet sterilization, and collecting crushed particles to obtain a solid fermentation substrate; then weighing 4700mL of deionized water, 400g of starch granules and 150g of sodium nitrate respectively, placing the materials in a stirrer, stirring and dispersing at room temperature, and collecting fermentation nutrient solution; spraying a fermentation nutrient solution onto the surface of a solid fermentation substrate, controlling the water content of the solid fermentation substrate to be 55%, inoculating trichoderma harzianum into the solid fermentation substrate in a dispersed manner according to the inoculation mass concentration of 7.5g/kg, culturing the trichoderma harzianum by using a freshness protection package bag with small holes filled, fermenting and culturing the trichoderma harzianum at 27 ℃ for 145 hours, opening the packaging bag and naturally drying the trichoderma harzianum for 27 hours after the fermentation culture is finished, stirring and mixing the air-dried fermentation product and sterile water according to the mass ratio of 1:9, filtering, collecting spore suspension, filtering for the second time and naturally drying the spore suspension to obtain dry spore powder;
culturing and collecting a bacillus agent: respectively weighing 1600g of corn flour, 1.70g of composite powder formed by mixing yeast powder, soybean meal and the like in mass, 80g of sodium chloride, 2g of manganese sulfate, 20g of dipotassium hydrogen phosphate and 11L of water, stirring, mixing, performing ultraviolet sterilization treatment, and collecting to obtain a liquid culture medium; respectively taking bacillus amyloliquefaciens, bacillus mucilaginosus and bacillus megaterium, respectively inoculating the bacillus amyloliquefaciens, the bacillus mucilaginosus and the bacillus megaterium into a liquid culture medium according to the inoculation volume ratio of 6%, performing heat preservation fermentation treatment at 38 ℃ for 50 hours, collecting fermentation liquor, performing centrifugal separation at 1500r/min, collecting lower-layer sediment, performing vacuum freeze drying, and mixing 20g of vacuum freeze-dried bacillus amyloliquefaciens, 20g of bacillus mucilaginosus and 20g of bacillus megaterium to obtain bacillus strains;
preparation of a culture medium carrier: weighing 4700g of diatomite, 1750g of grass carbon and 1250g of bran respectively, performing jet milling, and sieving with a 500-mesh sieve to obtain a culture medium carrier;
preparing a complex microbial inoculum: stirring and mixing 50g of dry spore powder, 30g of bacillus strain and 150g of culture medium carrier, preserving heat and drying at 37 ℃, crushing and sieving with a 200-mesh sieve to obtain a composite microbial inoculum;
preparing a coating modifying solution: stirring and mixing 100g of humic acid, 300mL of 1% ammonia water in mass fraction and 1000mL of deionized water, collecting the mixed solution, placing the mixed solution in a rotary evaporation device, magnetically stirring and mixing for 2 hours at 115 ℃ to obtain a mixed solution, rotationally evaporating to dryness, and collecting dry particles; weighing 4700mL of deionized water, 1250g of acrylamide and 400g of dry particles respectively, placing the deionized water, the 1250g of acrylamide and the dry particles into a reaction container, stirring and mixing the mixture, dropwise adding an ammonium persulfate solution with the mass fraction of 1% into the reaction container under the nitrogen atmosphere, controlling the dropwise adding amount to be 1% of the mass of the acrylamide, after the dropwise adding is finished, carrying out heat preservation reaction for 2 hours under the nitrogen atmosphere at 77 ℃, standing and cooling the mixture to room temperature to obtain a coating modification solution;
preparation of a microbial inoculum carrier: taking attapulgite, placing the attapulgite in a muffle furnace, calcining at 300 ℃ for 2h, standing and cooling to room temperature to obtain a modified attapulgite matrix, stirring and mixing 100mL of coating modification liquid and 1500g of the modified attapulgite matrix, stirring at 57 ℃ and 6MPa for 2h, filtering after the pressure stirring is finished, taking a filter cake, placing the filter cake at 105 ℃ for drying for 2h, collecting dried particles and grinding, and sieving with a 200-mesh sieve to obtain a microbial inoculum carrier;
preparing an alkaline soil remediation microbial inoculum: stirring and mixing 10g of the composite microbial inoculum, 20g of the mixed biochemical fulvic acid and ulmic acid with equal mass and 20g of a microbial inoculum carrier, and grinding and dispersing in a mortar to prepare the alkaline soil remediation microbial inoculum.
Example 3
Culturing and collecting trichoderma harzianum inoculum: naturally drying 500g of wood, 150g of bran and 300g of soybean meal, crushing, performing ultraviolet sterilization, and collecting crushed particles to obtain a solid fermentation substrate; respectively weighing 5000mL portions of deionized water, 300g of starch granules and 100g of sodium nitrate, placing the deionized water, the 300g of starch granules and the 100g of sodium nitrate into a stirrer, stirring and dispersing at room temperature, and collecting fermentation nutrient solution; spraying a fermentation nutrient solution onto the surface of a solid fermentation substrate, controlling the water content of the solid fermentation substrate to be 60%, then dispersedly inoculating trichoderma harzianum into the solid fermentation substrate according to the inoculation mass concentration of 7.5g/kg, culturing by using a freshness protection bag packed with small holes, fermenting and culturing for 150h at 28 ℃, after the fermentation culture is completed, opening the packaging bag and naturally drying for 30h, stirring and mixing 100g of air-dried fermentation product with 1000mL of sterile water, filtering, collecting spore suspension, secondarily filtering and naturally drying to obtain dry spore powder;
culturing and collecting a bacillus agent: respectively weighing 1800g of corn flour, 200 parts of composite powder formed by mixing yeast powder, soybean meal and the like in mass, 100g of sodium chloride, 3g of manganese sulfate, 30g of dipotassium hydrogen phosphate and 1.2L of water according to the parts by weight, stirring, mixing, performing ultraviolet sterilization treatment, and collecting to obtain a liquid culture medium; respectively taking bacillus amyloliquefaciens, bacillus mucilaginosus and bacillus megaterium, respectively inoculating the bacillus amyloliquefaciens, the bacillus mucilaginosus and the bacillus megaterium into a liquid culture medium according to the inoculation volume ratio of 6%, performing heat preservation fermentation treatment at 38 ℃ for 50 hours, collecting fermentation liquor, performing centrifugal separation at 1500r/min, collecting lower-layer sediment, performing vacuum freeze drying, and mixing 20g of vacuum freeze-dried bacillus amyloliquefaciens, 20g of bacillus mucilaginosus and 20g of bacillus megaterium to obtain bacillus strains;
preparation of a culture medium carrier: respectively weighing 5000g of diatomite, 2000g of turf and 1500g of bran, carrying out jet milling, and sieving with a 500-mesh sieve to obtain a culture medium carrier;
preparing a complex microbial inoculum: stirring and mixing 50g of dry spore powder, 30g of bacillus strain and 150g of microbial inoculum carrier, keeping the temperature and drying at 40 ℃, crushing, and sieving with a 200-mesh sieve to obtain a composite microbial inoculum;
preparing a coating modifying solution: stirring and mixing 100g of humic acid, 300mL of 1% ammonia water in mass fraction and 1000mL of deionized water, collecting the mixed solution, placing the mixed solution in a rotary evaporation device, magnetically stirring and mixing for 3 hours at 120 ℃, obtaining the mixed solution, placing the mixed solution in the rotary evaporation device, rotatably evaporating to dryness, and collecting dry particles; respectively weighing 5000g of deionized water, 1500g of acrylamide and 500g of dry particles, placing the deionized water, the 1500g of acrylamide and the 500g of dry particles into a reaction container, stirring and mixing, dropwise adding an ammonium persulfate solution with the mass fraction of 1% into the reaction container under the nitrogen atmosphere, controlling the dropwise adding amount to be 1% of the mass of the acrylamide, after the dropwise adding is finished, carrying out heat preservation reaction for 3 hours under the nitrogen atmosphere at the temperature of 80 ℃, standing and cooling to room temperature to obtain a coating modification solution;
preparation of a microbial inoculum carrier: taking attapulgite, placing the attapulgite in a muffle furnace, calcining at 350 ℃ for 3h, standing and cooling to room temperature to obtain a modified attapulgite matrix, stirring and mixing 100mL of coating modification liquid and 1500g of the modified attapulgite matrix, stirring at 60 ℃ and 8MPa for 3h, filtering after the pressure stirring is finished, taking a filter cake, placing the filter cake at 110 ℃ for drying for 3h, collecting dried particles, grinding, and sieving with a 200-mesh sieve to obtain a microbial inoculum carrier;
preparing an alkaline soil remediation microbial inoculum: stirring and mixing 10g of the composite microbial inoculum, 20g of the mixed biochemical fulvic acid and ulmic acid with equal mass and 20g of a microbial inoculum carrier, and grinding and dispersing in a mortar to prepare the alkaline soil remediation microbial inoculum.
Example 4
In example 4, the uncalcined and coated modified attapulgite is used as the microbial inoculum carrier, and other conditions and component proportions are the same as those in example 1.
Example 5
In example 5, calcined and modified attapulgite is used as a microbial inoculum carrier, and other conditions and component proportions are the same as those in example 1.
Example 6
In example 6, the attapulgite which is not calcined and modified but coated and modified is used as the microbial inoculum carrier, and other conditions and component proportions are the same as those in example 1.
Example 7
In example 7, polyacrylamide is directly used to coat the calcined attapulgite to prepare the microbial inoculum carrier, and other conditions and component proportions are the same as those in example 1.
Examples 8 to 14
In examples 8 to 14, the proportion of each bacillus and trichoderma harzianum is optimized to prepare the composite microbial inoculum, other conditions and component proportions are the same as those in example 3, and the specific proportion table is shown in table 1 below:
table 1 optimization of the ratio of Bacillus and Trichoderma harzianum in examples 8-14
Performance testing tests are respectively carried out on the repairing effect of the alkaline soil repairing microbial inoculum prepared in the embodiment 1-14, the testing place is a double-village living vegetable greenhouse land (suining county cultivated land quality protection station) in the urban county of suining county in xu city, Jiangsu province, and the soil agro-chemical property of the land is shown in the following table 2:
TABLE 2 soil agro-chemical traits Table
| Test point | pH | Organic matter (g/kg) | Total nitrogen (g/kg) | Quick-acting phosphorus (ppm) | Type of soil | Quick-acting potassium (ppm) |
| Double Yingcun | 8.2 | 18.61 | 1.78 | 75.3 | Yellow moisture soil | 135.9 |
Applying the basic soil remediation microbial inoculum once every 30 days by adopting a conventional application mode of applying fertilizer and 50 kg/mu of the basic soil remediation microbial inoculum, picking the first time after 60 days after planting the nodulized lettuce, applying the basic soil remediation microbial inoculum for applying for treating every 60 days for the second planting, and detecting the agronomic characters of soil agricultural products after the second planting, wherein the detection results are shown in the following tables 4-5.
Detection method/test method
And (3) soil agro-chemical characters after planting for 60 days:
the pH of the soil adopts a suspension with a water-soil ratio of 5:1, and is measured by a pH meter; measuring the soil triphase ratio and the soil total porosity by using a soil triphase meter; the soil nutrients were measured using a continuous flow analyzer, and the results are shown in table 3 below.
Agricultural product agronomic characters after 60 days of planting:
the height, the weight and the leaf number of lettuce plants produced in one year are respectively counted, meanwhile, the content of vitamin C, soluble total sugar and nitrate is tested, the yield per mu is counted, and the detection results are shown in the following tables 4-5.
The specific detection results are shown in the following tables 2-5:
TABLE 3 soil property test tables for examples 1 to 14
TABLE 4 examples 1-14 agronomic traits of agricultural products
TABLE 5 examples 1-14 agronomic traits for second crop planting
The performance test comparison in reference to tables 2-5 can find that: comparing the performances of the embodiments 1 to 3, the pH is obviously reduced in the embodiments 1 to 3, and the higher the pressure is when the modified attapulgite matrix and the coating modifying solution are stirred and mixed in the embodiments 1 to 3, the lower the pH value of the alkaline soil is after the prepared microbial inoculum is used, which shows that by increasing the mixing pressure of the coating modifying solution and the modified attapulgite, the microbial inoculum can be more deeply loaded in the pores inside the modified attapulgite, and by mutual association between the molecular groups and water molecules, a good propagation environment is provided for the microbes in the microbial inoculum, so that the service life of the microbial inoculum material is prolonged, and the alkaline soil is finally effectively improved.
Comparing the performances of the embodiment 1 and the embodiment 4, in the preparation process of the embodiment 4, the attapulgite which is not calcined and is coated and modified is used as the microbial inoculum carrier, although the alkaline soil can be effectively repaired, compared with the embodiment 1, the repair performance of the attapulgite is poorer than that of the embodiment 1, which shows that the attapulgite which is calcined and modified is coated and modified by the coating liquid, the loading effect of the loaded microbial inoculum is improved, the service life of the microbial inoculum material is prolonged, and finally the alkaline soil is effectively improved and the stability of the alkaline soil is improved.
Comparing the performances of the embodiment 1, the embodiment 4 and the embodiment 5, the calcined and modified attapulgite is used as the microbial inoculum carrier in the embodiment 5, although the alkaline soil can be effectively repaired, the repairing performance is reduced compared with the embodiment 1, but the repairing performance is better than that of the embodiment 4 compared with the embodiment 4, which shows that the calcination modification treatment of the attapulgite adopted by the application enables the microbial inoculum to be effectively loaded in the pores inside the attapulgite, provides a good propagation environment for the microorganisms in the microbial inoculum, thereby prolonging the service life of the microbial inoculum material and finally effectively improving the alkaline soil.
Comparing the performances of the embodiment 1, the embodiment 4 and the embodiment 6, the embodiment 6 adopts the attapulgite which is not calcined and modified but is coated and modified as the microbial inoculum carrier, and as can be seen from tables 2 to 4, the performance of the embodiment 6 is reduced compared with that of the embodiment 1, but the performance is improved compared with that of the embodiment 4, which shows that the coated attapulgite has good structural performance, the service life of the microbial inoculum material is prolonged, and finally the alkaline soil is effectively improved.
Comparing the performances of the embodiment 1, the embodiment 4 and the embodiment 7, the embodiment 7 adopts polyacrylamide to directly coat and modify calcined attapulgite to prepare a microbial inoculum carrier, and as can be seen from tables 2 to 4, the embodiment 6 has performance lower than that of the embodiment 1, but the embodiment 6 also has performance higher than that of the embodiment 4, but as can be seen from table 4, the durability is not good, which shows that the ammonium humate modified alkaline soil adopted by the application can reduce the soil alkalinity, form a good microbial propagation environment and improve the initial improvement efficiency of the microbial inoculum.
Comparing the example 1 with the examples 8 to 14, it can be found from tables 3 to 5 that the pH of the treated soil is reduced when the addition amount of the bacillus megaterium reaches the maximum ratio under the condition that the other components except the ratio of the bacillus megaterium and the process conditions are not changed, which indicates that the treatment effect of the microbial agent can be effectively improved by increasing the ratio of the bacillus megaterium, and then, the observation of tables 3 to 5 shows that the total salt content of the soil is remarkably reduced when the ratio of the bacillus mucilaginosus is higher, which indicates that the texture and the micro-ecological environment in the soil can be effectively improved by increasing the ratio of the bacillus mucilaginosus in the microbial agent, and finally, when the ratio of the bacillus amyloliquefaciens is higher, the yield of the planting product can be remarkably increased, which indicates that the yield of the planting product can be remarkably increased by increasing the ratio of the bacillus amyloliquefaciens, the yield of planting products can be improved, and in conclusion, the alkaline soil can be effectively improved by increasing the specific gravity of each bacillus, the soil fertilizer efficiency can be effectively improved, and the planting yield of crops can be increased.
Comparative example
Comparative examples 1 to 3
In comparative examples 1 to 3, calcined diatomite is used for preparing the microbial agent carrier, and the conditions and the component proportion are the same as in comparative examples 1 to 3 corresponding to examples 1 to 3 respectively.
Comparative examples 4 to 6
In comparative examples 4 to 6, polyvinyl alcohol is used for coating, modifying and calcining attapulgite to prepare the microbial inoculum carrier, and the conditions and the component proportions are the same as in comparative examples 4 to 6 respectively corresponding to examples 1 to 3.
Comparative examples 7 to 9
In comparative examples 7-9, microbial agents were prepared by mass compounding of only Paenibacillus mucilaginosus and Trichoderma harzianum, and the conditions and component ratios were the same as in comparative examples 7-9 corresponding to examples 1-3, respectively.
In the comparative examples 10 to 12, the microbial agent is prepared by only adopting mass compounding of the bacillus mucilaginosus, the bacillus amyloliquefaciens, the bacillus megaterium and the like, and the conditions and the component ratio are the same as those in the comparative examples 10 to 12 respectively corresponding to the examples 1 to 3.
In the comparative examples 13 to 15, the microbial agent is prepared by only adopting mass compounding of bacillus amyloliquefaciens, trichoderma harzianum and the like, and the conditions and the component ratio are the same as those of the comparative examples 13 to 15 respectively corresponding to the examples 1 to 3.
In the comparative examples 16 to 18, the microbial agent is prepared by only adopting mass compounding of bacillus megatherium, trichoderma harzianum and the like, and the conditions and the component ratio are the same as those in the comparative examples 16 to 18 respectively corresponding to the examples 1 to 3.
In the comparative examples 19 to 21, the microbial agent is prepared by only adopting mass compounding of bacillus megatherium, paenibacillus mucilaginosus, trichoderma harzianum and the like, and the conditions and the component proportion are the same as in the comparative examples 19 to 21 respectively corresponding to the examples 1 to 3.
In the comparative examples 22 to 24, the microbial agent is prepared by only adopting mass compounding of bacillus megatherium, bacillus amyloliquefaciens, trichoderma harzianum and the like, and the conditions and the component proportion are the same as in the comparative examples 22 to 24 respectively corresponding to the examples 1 to 3.
In the comparative examples 25 to 27, the microbial agent is prepared by only adopting mass compounding of the paenibacillus jelly, the bacillus amyloliquefaciens and the trichoderma harzianum, and the like, and the conditions and the component proportion are the same as in the comparative examples 25 to 27 respectively corresponding to the examples 1 to 3.
Comparative examples 28 to 30
The comparative examples 28 to 30 are prepared without adding biochemical organic acid when preparing the remediation microbial inoculum, and are the same as the comparative examples 28 to 30 respectively corresponding to the examples 1 to 3 in the rest conditions and component proportions.
Performance test
Respectively testing the repairing effect of the alkaline soil repairing microbial inoculum prepared in the proportion of 1-12, wherein the testing place is a double-operation village lettuce greenhouse land (suining county cultivated land quality protection station) in the king county of suining county and county of Xuzhou city, Jiangsu province, applying the alkaline soil repairing microbial inoculum once every 30 days by adopting a conventional fertilization and alkaline soil repairing microbial inoculum applying mode, picking the first time after 60 days after planting lettuce nodulation, applying the applying amount of the alkaline soil repairing microbial inoculum for processing the second time after planting every 60 days, and detecting the agronomic characters of soil agricultural products after the second time of planting.
Detection method/test method
And (3) detecting the soil agroforestry traits after 60 days of planting: the pH of the soil adopts a suspension with a water-soil ratio of 5:1, and is measured by a pH meter; measuring the soil triphase ratio and the soil total porosity by using a soil triphase meter; the soil nutrients were measured using a continuous flow analyzer, and the results are shown in table 6 below.
Agricultural product agronomic characters after 60 days of planting:
the height, weight and leaf number of the lettuce plants produced in one year are respectively counted, the content of vitamin C, soluble total sugar and nitrate is tested, the yield per mu is counted, and the detection results are shown in the following table 7.
The specific detection results are shown in the following tables 6-7:
TABLE 6 soil property test tables for comparative examples 1 to 12
TABLE 7 agricultural product agronomic performance of comparative examples 1-12
The comparison of the performance tests in reference to tables 6-7 can be found as follows:
compared with the comparative examples 1-3 and the examples 1-3, the calcined diatomite is adopted in the formula of the comparative example to prepare the microbial inoculum carrier, although the performance of the microbial inoculum carrier is poorer than that of the examples 1-3, the microbial inoculum carrier also has a good repairing effect, which shows that in the repairing microbial inoculum adopted in the application, microorganisms play a huge role in the repairing effect of the microbial inoculum carrier and the microorganisms, namely, the alkaline soil can be effectively improved by adopting a microbial inoculum scheme of multi-strain combination.
Compared with the examples 1-3, the comparative examples 4-6 of the application adopt the polyvinyl alcohol to coat, modify and calcine the attapulgite to prepare the microbial inoculum carrier, the repairing performance of the carrier is in a descending trend, so that the polyacrylamide adopted by the application can effectively maintain the structure of soil aggregates, and through mutual association between molecular groups and water molecules, a good propagation environment is provided for microorganisms in the microbial inoculum, so that the service life of the microbial inoculum material is prolonged, and finally the alkaline soil is effectively improved.
Comparing the comparative examples 7-29 with the examples 1-3 in sequence, respectively, in the preparation process of the comparative examples 7-27, only any two or any three of bacillus megatherium, paenibacillus mucilaginosus, bacillus amyloliquefaciens and trichoderma harzianum are adopted for equal mass compounding to prepare the microbial agent, and in the using scheme, the using effect of any two or any three of the equal mass compounded microbial agents is the worst, and the repairing effect of any three of the microbial agents is the second lowest, so that the repairing effect of the microbial agent can be reduced by adopting the scheme of combining any two or any three of the microbes, the composition components of the microbial agent are optimized, the basic soil is improved by adopting the scheme of combining multiple strains of microbial agents, and the defect of a single-strain microbial agent is improved.
Comparing the comparative examples 10-12 with the examples 1-3, the comparative examples 10-12 are prepared without adding biochemical organic acid when preparing the remediation microbial inoculum, so that the remediation effect is slightly different from the examples 1-3, which shows that the organic acid is adopted for improvement, the alkaline components in the soil can be primarily neutralized, a good environment is provided for the propagation of microorganisms, the improvement life is prolonged, and the alkaline soil is finally and effectively improved.
The present embodiment is only for explaining the present application, and it is not limited to the present application, and those skilled in the art can make modifications of the present embodiment without inventive contribution as needed after reading the present specification, but all of them are protected by patent law within the scope of the claims of the present application.
Claims (3)
1. The alkaline soil remediation microbial inoculum is characterized by comprising the following substances in parts by weight:
25-30 parts of a microbial inoculum carrier;
10-15 parts of a composite microbial inoculum;
25-30 parts of biochemical organic acid;
the composite microbial agent comprises Bacillus amyloliquefaciens, Bacillus mucilaginosus, Bacillus megatherium and Trichoderma harzianum which are mixed according to the mass ratio of 1-2: 5;
the compound microbial inoculum is prepared by the following method:
s1, preparing mixed strains: respectively activating and culturing bacillus amyloliquefaciens, bacillus mucilaginosus, bacillus megaterium and trichoderma harzianum spore powder according to the formula, and collecting thalli to obtain mixed strains;
s2, preparation of a culture medium carrier: respectively taking diatomite, turf and bran, and carrying out jet milling and sieving to obtain a culture medium carrier;
s3, preparing a complex microbial inoculum: stirring and mixing the mixed strain and a culture medium carrier, keeping the temperature at 35-40 ℃, drying, and crushing to obtain a composite microbial inoculum;
the microbial inoculum carrier is prepared by the following method:
(1) calcining attapulgite at 250-300 ℃, standing and cooling to obtain a modified attapulgite matrix;
(2) stirring and mixing the coating modification solution and the modified attapulgite matrix, carrying out heat preservation and pressurization stirring treatment at 55-60 ℃ and 5-8 MPa, filtering, drying a filter cake, collecting dry particles, and grinding to obtain a microbial inoculum carrier;
the coating modifying solution is prepared by the following method:
(1) stirring and mixing humic acid, ammonia water with the mass fraction of 1% and deionized water according to the mass ratio of 1: 3-5: 10-15, collecting a mixed solution, rotating to evaporate to dryness, collecting dry particles, and mixing the dry particles according to the mass ratio of 9-10: 3-5: 1 stirring and mixing deionized water, acrylamide and dry particles;
(2) in a nitrogen atmosphere, taking an ammonium persulfate solution as an initiator, reacting for 2-3 h at 75-80 ℃ in a nitrogen atmosphere in a heat preservation manner, standing and cooling to room temperature to obtain a coating modification solution;
the biochemical organic acid consists of biochemical fulvic acid and fulvic acid which are mixed by equal mass;
the total effective viable count of the alkaline soil remediation microbial inoculum is not less than 15.0 hundred million cfu/g, and the rate of mixed bacteria of the alkaline soil remediation microbial inoculum is not more than 20%.
2. The alkaline soil remediation microbial inoculum of claim 1, wherein the size of the sieved mesh of step S2 is 500 meshes.
3. The method for preparing an alkaline soil remediation microbial inoculum as claimed in claim 1 or claim 2, wherein the step of preparing the alkaline soil remediation microbial inoculum comprises: and stirring and mixing the composite microbial inoculum, the biochemical organic acid and a microbial inoculum carrier according to a formula, and grinding and dispersing in a mortar to prepare the alkaline soil remediation microbial inoculum.
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