CN113150788A - Preparation method and application of seaweed oligosaccharide composite microbial material for preventing soil hardening - Google Patents
Preparation method and application of seaweed oligosaccharide composite microbial material for preventing soil hardening Download PDFInfo
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- C09K17/14—Soil-conditioning materials or soil-stabilising materials containing organic compounds only
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Abstract
The invention provides a preparation method and application of a seaweed oligosaccharide composite microbial material for preventing soil hardening, and belongs to the technical field of soil treatment and restoration. The preparation process of the seaweed oligosaccharide composite microbial material comprises the following steps: firstly, preprocessing pine bark, then soaking the pine bark in a seaweed oligosaccharide solution to enable the seaweed oligosaccharide solution to be attached to the surface of the bark or to be soaked into a pore channel of the bark, and then soaking the composite material into a phosphate solubilizing bacterium liquid to enable phosphate solubilizing bacteria to be fully adsorbed on the composite material. The seaweed oligosaccharide composite microbial material prepared by the invention has certain air permeability, water retention and fertility increasing functions, further effectively activates a soil layer and enhances the permeability of rich water. Meanwhile, the phosphate solubilizing bacteria used in the invention can improve the effective phosphorus content of soil and prevent soil hardening.
Description
Technical Field
The invention belongs to the technical field of soil treatment and restoration, and particularly relates to a preparation method and application of a seaweed oligosaccharide composite microbial material for preventing soil hardening.
Background
China is a big agricultural country, and agricultural production activities have a history of thousands of years. With the continuous progress of farming, various farmlands in China have soil hardening phenomena of different degrees, the situation of dry North China is particularly serious, the soil hardening causes the reduction of water retention, fertilizer retention and permeability, and normal farming activities can not be carried out, thus becoming an important factor for restricting the sustainable development of rural productivity gradually. The soil is of a granular structure, and when water falls, water in the soil flows out of holes to cause dehydration; the other is that phosphate fertilizer is used for a long time in China, but the utilization rate of the phosphate fertilizer is low, and redundant phosphorus element can react with metal ions in soil in the form of phosphate ions to generate insoluble phosphate (such as Mg)3(PO4)2) The structure of the soil is influenced, and the risk of soil hardening is increased.
The current common methods for treating soil hardening comprise sprinkling irrigation, straw returning and the like. However, these methods only consider the physical structure and chemical composition of soil, and are not effective in studying the role of microorganisms in treating soil hardening. For example, chinese patent 201711372272.4 discloses a method for controlling soil compaction using deep ploughing, rotary tillage and organic fertilizers. Although the method solves the problem of soil hardening to a certain extent, the repair is not thorough and the recurrence is easy. Other disclosed methods, such as those disclosed in patents 201810226918.6, 201810081136.8, 201610754615.2, etc., also simply combine several microorganisms to form a combined microbial preparation, which is then used with some agricultural waste, and do not provide a stable and efficient treatment solution.
The phosphate solubilizing bacteria are special microbial functional groups, can convert refractory phosphorus in soil into soluble phosphorus which can be absorbed by plants, and can activate the insoluble phosphorus in the soil by secreting various enzymes and organic acids, so that the effective phosphorus content of the soil is increased, and the growth and development of the plants are promoted. The stability of the microorganisms is susceptible to the soil or the weather environment, and if several phosphate solubilizing bacteria are directly and simply put into the soil, the effect is difficult to be effectively exerted.
Disclosure of Invention
In view of the above, the present invention aims to provide an alginate-oligosaccharide composite microbial material for preventing soil hardening, a preparation method and an application thereof, wherein the alginate-oligosaccharide composite microbial material promotes the circulation of phosphorus in soil by using the metabolic process of phosphate solubilizing bacteria, and improves the water retention of soil by using an alginate-oligosaccharide solution.
In order to achieve the above object, the present invention provides an algal oligosaccharide complex microbial material for preventing soil hardening; the effective viable bacteria concentration of the phosphate-solubilizing bacterium liquid is 107～1010cfu/ml。
Preferably, the mass concentration of the seaweed oligosaccharide solution is 0.5-2.0%, and more preferably 1.2%; the concentration of the hydrochloric acid solution is 1-3 mol/L, and more preferably 2 mol/L.
Preferably, the phosphate solubilizing bacterium liquid comprises staphylococcus pasteurii, pseudomonas and chromobacterium violaceum, and the effective viable bacteria concentration ratio of the staphylococcus pasteurii to the pseudomonas to the chromobacterium violaceum is 2.5-3.5: 2-3: 1, and more preferably 3:2.5: 1.
The invention provides a preparation method of the seaweed oligosaccharide composite microbial material, which comprises the following steps:
(1) cutting pine bark into certain size, and performing microwave treatment;
(2) adding the pine bark subjected to microwave treatment into a hydrochloric acid solution, heating to 50-80 ℃, reacting for 1-3 h, filtering, washing with distilled water until the filtrate is neutral, and drying the pine bark in a blast drying oven at 25 ℃ to obtain clean bark;
(3) preparing the seaweed oligosaccharide solution, soaking clean barks in the oligosaccharide solution at room temperature, taking out after 24-30 h, slightly washing with deionized water for 2-4 times, and naturally drying in the air;
(4) and soaking the soaked pine bark in the phosphate-solubilizing bacteria liquid for 24-30 h.
The invention also provides application of the seaweed oligosaccharide composite microbial material in preventing soil hardening.
The application preferably comprises the following steps: the seaweed oligosaccharide composite microbial material is uniformly thrown into hardened soil, and the throwing amount of the material is 0.2-0.4 kg per mu, and more preferably 0.3kg per mu.
Compared with the prior art, the invention has the following advantages:
the invention provides a seaweed oligosaccharide composite microbial material for preventing soil hardening, which is stable in property, and low in cost, and pine bark used as the material is waste. The composite microbial material is applied to hardened soil, and can play a good role in promoting phosphorus circulation, stabilizing soil aggregate structure, improving effective phosphorus content and water-retaining property of soil, repairing and preventing soil hardening.
Detailed Description
The invention provides a seaweed oligosaccharide composite microbial material for preventing soil hardening.
The composite microbial material is characterized in that a seaweed oligosaccharide solution is attached to the pore diameter of pine bark pretreated by a hydrochloric acid solution, and the formed composite material is immersed in a phosphate solubilizing bacteria liquid to form the seaweed oligosaccharide composite microbial material. The mass concentration of the seaweed oligosaccharide solution is preferably 0.5-2.0%, and more preferably 1.2%; the concentration of the hydrochloric acid solution is preferably 2 mol/L. The sources of the algal oligosaccharide solution and hydrochloric acid solution are not particularly limited in the present invention, and any commercially available product that is conventional in the art may be used.
In the invention, the effective viable bacteria concentration of the phosphate solubilizing bacteria liquid is 107～1010cfu/ml, preferably 109cfu/ml; in the present invention, the phosphate solubilizing bacteria liquid preferably includes Staphylococcus pasteurii (Staphylococcus pasteurii), Pseudomonas fluorescens Migula (Pseudomonas fluorescens), and Chromobacterium violaceum (Chromobacterium violaceum). The effective viable bacteria concentration ratio of the staphylococcus pasteurii to the pseudomonas to the chromobacterium violaceum is preferably 2.5-3.5: 2-3: 1, and more preferably 3:2.5: 1. The source of the phosphate solubilizing bacteria is not particularly limited, and the bacteria with the phosphate solubilizing function can be any bacteria in the field. In the examples of the present invention, the ATCC number of Staphylococcus pasteurii is 51129, the ATCC number of Pseudomonas is 13525, andthe chromobacterium violaceum has ATCC number 31532 and may be purchased from strain depository.
The invention provides a preparation method of the microbial material, which comprises the following steps:
(1) cutting pine bark into certain size, and performing microwave treatment;
(2) adding the pine bark subjected to microwave treatment into a hydrochloric acid solution, heating to 50-80 ℃, reacting for 1-3 h, filtering, washing with distilled water until the filtrate is neutral, and drying the pine bark in a blast drying oven at 25 ℃ to obtain clean bark;
(3) preparing the seaweed oligosaccharide solution, soaking clean barks in the oligosaccharide solution at room temperature, taking out after 24-30 h, slightly washing for 3 times by using deionized water, and naturally drying in the air;
(4) and soaking the soaked pine bark in the phosphate-solubilizing bacterium liquid.
The invention cuts the pine bark into the same size and then carries out microwave treatment. The pine bark has the length of 9-10 mm, the width of 9-10 mm and the thickness of 2-3 mm; the microwave power is preferably 500-900W, and more preferably 700W; the time is preferably 2-5 min.
After the microwave treatment, the pine bark is added into a hydrochloric acid solution with the concentration of 1-3 mol/L, the mixture is heated to 50-80 ℃, the reaction is carried out for 1-3 hours, the mixture is filtered, the filtrate is washed by distilled water until the filtrate is neutral, and the filtrate is dried in an air blast drying oven at the temperature of 25 ℃ to obtain the clean pine bark. The concentration of the hydrochloric acid solution is preferably 1-3 mol/L, and more preferably 2 mol/L; the reaction time is preferably 1-3 h, and more preferably 2 h; the number of times of washing with distilled water is preferably 2 to 4, more preferably 3.
The preparation method comprises the steps of preparing the seaweed oligosaccharide solution, soaking clean barks in the oligosaccharide solution at room temperature, taking out after 24-30 h, slightly washing for 2-4 times by using deionized water, and naturally drying. The mass concentration of the seaweed oligosaccharide is preferably 0.5-2%, and more preferably 1.2%; the number of washing times is preferably 2 to 4, more preferably 3.
After washing with deionized water, the invention soaks the pine bark in the phosphate solubilizing bacteria liquid. The phosphate solubilizing bacteria liquid preferably includes Staphylococcus pasteurii (Staphylococcus pasteurii), Pseudomonas fluorescens Migula (Pseudomonas fluorescens), and Chromobacterium violaceum (Chromobacterium violacea). The effective viable bacteria concentration ratio of the staphylococcus pasteurii to the pseudomonas to the chromobacterium violaceum is preferably 2.5-3.5: 2-3: 1, and more preferably 3:2.5: 1; the soaking time is preferably 24-30 h.
The invention also provides application of the composite microbial material in preventing soil hardening. In the invention, the application steps are as follows: and uniformly throwing the composite microbial material into hardened soil, wherein the throwing amount of the material is 0.2-0.4 kg per mu, and more preferably 0.3kg per mu.
The hardened soil treated by the method has the advantages of increased effective phosphorus content, reduced soil volume weight, increased soil porosity and controlled hardening phenomenon.
The following will explain in detail a microbial material for preventing soil hardening, a method for preparing the same, and applications thereof, provided by the present invention, with reference to examples and comparative examples, which should not be construed as limiting the scope of the present invention.
Example 1
(1) Preparing bacterial suspensions of Staphylococcus pasteruri (bacterium a), pseudomonas fluorescens Migula (bacterium b) and pseudomonas violacea (bacterium c), and respectively determining effective viable bacteria concentration to be bacterium a and 5-8 multiplied by 109cfu/ml; bacterium b, 3-5 × 109cfu/ml; bacterium c, 2-4X 109cfu/ml. Then mixing the bacterium a suspension, the bacterium b suspension and the bacterium c suspension according to the volume of 3:2.5:1 to obtain a composite bacterium liquid (the total concentration of effective viable bacteria is about 3-7 multiplied by 10)9cfu/ml)。
(2) Cutting the pine bark into certain size, and then carrying out microwave treatment for 2min under the power of 700W (the length of the pine bark is 9-10 mm, the width is 9-10 mm, and the thickness is 2-3 mm). Adding microwave treated cortex Pini into 2mol/L hydrochloric acid solution, heating to 50 deg.C, reacting for 2 hr, filtering, washing with distilled water until the filtrate is neutral, and drying at 25 deg.C in forced air drying oven for 60 hr to obtain clean cortex Pini.
(3) Preparing 1.2% of the seaweed oligosaccharide solution, soaking clean barks in the oligosaccharide solution at room temperature, taking out after 24h, slightly washing for 3 times by using deionized water, and naturally drying for 2h to obtain the composite material.
(4) And (3) soaking the composite material in the composite bacterial liquid for 24 hours to obtain the composite microbial material. Then 0.3kg of material is evenly thrown into the soil of each mu of non-planted crops, phosphate solubilizing bacteria are utilized to promote the phosphorus circulation, the granular structure of the soil is stabilized, the effective phosphorus content is improved, the effective phosphorus content of the soil is increased after two weeks, and the volume weight of the soil is 2.0g/cm3Reduced to 1.0g/cm3The porosity of the soil is increased from 35% to 56%, and the soil hardening phenomenon is controlled.
Example 2
(1) Preparing bacterial suspensions of Staphylococcus pasteruri (bacterium a), pseudomonas fluorescens Migula (bacterium b) and pseudomonas violacea (bacterium c), and respectively determining effective viable bacteria concentration to be bacterium a and 5-8 multiplied by 109cfu/ml; bacterium b, 2-4X 109cfu/ml; bacterium c, 1-2X 109cfu/ml. Then mixing the bacterium a suspension, the bacterium b suspension and the bacterium c suspension according to the volume of 2.5:2:1 to obtain a composite bacterium liquid (the total concentration of effective viable bacteria is about 3-6 multiplied by 10)9cfu/ml)。
(2) Cutting the pine bark into certain size, and then carrying out microwave treatment for 2min under the power of 700W (the length of the pine bark is 9-10 mm, the width is 9-10 mm, and the thickness is 2-3 mm). Adding microwave treated cortex Pini into 2mol/L hydrochloric acid solution, heating to 50 deg.C, reacting for 2 hr, filtering, washing with distilled water until the filtrate is neutral, and drying at 25 deg.C in forced air drying oven for 60 hr to obtain clean cortex Pini.
(3) Preparing 1.2% of the seaweed oligosaccharide solution, soaking clean barks in the oligosaccharide solution at room temperature, taking out after 24h, slightly washing for 3 times by using deionized water, and naturally drying for 2h to obtain the composite material.
(4) And (3) soaking the composite material in the composite bacterial liquid for 24 hours to obtain the composite microbial material. Then 0.3kg of material is evenly thrown into the soil of each mu of non-planted crops, phosphate solubilizing bacteria are utilized to promote the circulation of phosphorus, the granular structure of the soil is stabilized, the effective phosphorus content is improved,after two weeks, the available phosphorus content of the soil is increased, and the volume weight of the soil is increased from 2.0g/cm3Reduced to 1.2g/cm3The porosity of the soil is increased from 35% to 50%, and the soil hardening phenomenon is controlled.
Example 3
(1) Preparing bacterial suspensions of Staphylococcus pasteruri (bacterium a), pseudomonas fluorescens Migula (bacterium b) and pseudomonas violacea (bacterium c), and respectively determining effective viable bacteria concentrations to be bacterium a and 3-5 multiplied by 109cfu/ml; bacterium b, 3-5 × 109cfu/ml; bacterium c, 3-5 × 109cfu/ml. Then mixing the bacterium a suspension, the bacterium b suspension and the bacterium c suspension according to the volume of 2.5:3:1 to obtain a composite bacterium liquid (the total concentration of effective viable bacteria is about 3-5 multiplied by 10)9cfu/ml)。
(2) Cutting the pine bark into certain size, and then carrying out microwave treatment for 2min under the power of 700W (the length of the pine bark is 9-10 mm, the width is 9-10 mm, and the thickness is 2-3 mm). Adding microwave treated cortex Pini into 2mol/L hydrochloric acid solution, heating to 50 deg.C, reacting for 2 hr, filtering, washing with distilled water until the filtrate is neutral, and drying at 25 deg.C in forced air drying oven for 60 hr to obtain clean cortex Pini.
(3) Preparing 1.2% of the seaweed oligosaccharide solution, soaking clean barks in the oligosaccharide solution at room temperature, taking out after 24h, slightly washing for 3 times by using deionized water, and naturally drying for 2h to obtain the composite material.
(4) And (3) soaking the composite material in the composite bacterial liquid for 24 hours to obtain the composite microbial material. Then 0.3kg of material is evenly thrown into the soil of each mu of non-planted crops, phosphate solubilizing bacteria are utilized to promote the phosphorus circulation, the granular structure of the soil is stabilized, the effective phosphorus content is improved, the effective phosphorus content of the soil is increased after two weeks, and the volume weight of the soil is 2.0g/cm3Reduced to 1.3g/cm3The porosity of the soil is increased from 35% to 48%, and the soil hardening phenomenon is controlled.
Example 4
(1) Preparing bacterial suspensions of Staphyloccus pasteuri (bacterium a), Pseudomonas marginalis (bacterium b) and Chromobacterium violacea (bacterium c), and determining effective viable bacteria concentrations to be respectively bacterium a and 2-3 multiplied by 109cfu/ml; bacterium b, 3-5 × 109cfu/ml; bacterium c, 1-5X 109cfu/ml. Then mixing the bacterium a suspension, the bacterium b suspension and the bacterium c suspension according to the volume of 3.5:2:1 to obtain a composite bacterium liquid (the total concentration of effective viable bacteria is about 2-4 multiplied by 10)8cfu/ml)。
(2) Cutting the pine bark into certain size, and then carrying out microwave treatment for 2min under the power of 700W (the length of the pine bark is 9-10 mm, the width is 9-10 mm, and the thickness is 2-3 mm). Adding the pine bark subjected to microwave treatment into a 2mol/L hydrochloric acid solution, heating to 50-80 ℃, reacting for 2h, filtering, washing with distilled water until the filtrate is neutral, and drying the pine bark in a 25 ℃ forced air drying oven for 60h to obtain clean bark.
(3) Preparing 1.2% of the seaweed oligosaccharide solution, soaking clean barks in the oligosaccharide solution at room temperature, taking out after 24h, slightly washing for 3 times by using deionized water, and naturally drying for 2h to obtain the composite material.
(4) And (3) soaking the composite material in the composite bacterial liquid for 24 hours to obtain the composite microbial material. Then 0.3kg of material is evenly thrown into the soil of each mu of non-planted crops, phosphate solubilizing bacteria are utilized to promote the phosphorus circulation, the granular structure of the soil is stabilized, the effective phosphorus content is improved, the effective phosphorus content of the soil is increased after two weeks, and the volume weight of the soil is 2.0g/cm3Reduced to 1.5g/cm3The porosity of the soil is increased from 35% to 45%, and the soil hardening phenomenon is controlled.
The foregoing is only a preferred embodiment of the present invention, and it should be noted that, for those skilled in the art, various modifications and decorations can be made without departing from the principle of the present invention, and these modifications and decorations should also be regarded as the protection scope of the present invention.
The above description is only for the purpose of illustrating the preferred embodiments of the present invention, and the present invention is not limited to the illustrated embodiments, and all the modifications and equivalents of the embodiments may be made without departing from the spirit of the present invention.
Claims (8)
1. A preparation method of a seaweed oligosaccharide composite microbial material for preventing soil hardening is characterized by comprising the following steps:
(1) cutting pine bark into certain size, and performing microwave treatment;
(2) adding the pine bark subjected to microwave treatment into a hydrochloric acid solution, heating to 50-80 ℃, reacting for 1-3 h, filtering, washing with distilled water until the filtrate is neutral, and drying the pine bark in a blast drying oven at 25 ℃ to obtain clean bark;
(3) preparing a seaweed oligosaccharide solution, soaking clean barks in the seaweed oligosaccharide solution at room temperature, taking out after 24-30 h, slightly washing with deionized water for 2-4 times, and naturally drying in the air;
(4) and soaking the soaked pine bark in the phosphate-solubilizing bacteria liquid for 24-30 h, slightly washing the pine bark with deionized water for 2-4 times, and naturally drying the pine bark.
2. The method for preparing the algal oligosaccharide complex microbial material for preventing soil hardening according to claim 1, wherein the pine bark of step (1) has a length of 9mm to 10mm, a width of 9mm to 10mm and a thickness of 2mm to 3 mm; the concentration of the hydrochloric acid solution in the step (2) is 1-3 mol/L; the mass concentration of the seaweed oligosaccharide solution in the step (3) is 0.5-2%; the effective viable bacteria concentration of the phosphate-solubilizing bacteria liquid in the step (4) is 107～1010cfu/mL。
3. The method for preparing the algal oligosaccharide composite microbial material for preventing soil hardening according to claim 1, wherein the power of the microwave in the step (1) is 500-900W; the time is 2-5 min.
4. The method for preparing the algal oligosaccharide complex microbial material for preventing soil hardening according to claim 1, wherein the drying time in the step (2) is 48-72 h, and more preferably 60 h.
5. The preparation method of the algal oligosaccharide composite microbial material for preventing soil hardening as claimed in claim 1, wherein the phosphate solubilizing bacteria liquid in the step (4) comprises staphylococcus pasteurii, pseudomonas and chromobacterium violaceum, wherein the effective viable bacteria concentration ratio of the staphylococcus pasteurii to the chromobacterium violaceum is 2.5-3.5: 2-3: 1.
6. An algal oligosaccharide complex microbial material for preventing soil hardening prepared by the method according to any one of claims 1 to 5.
7. The algal oligosaccharide complex microbial material according to claim 6 for preventing soil hardening, which can be used for preventing soil hardening.
8. The application of the seaweed oligosaccharide composite microbial material for preventing soil hardening as claimed in claim 7, is characterized in that the application method comprises the following steps:
and uniformly throwing the microbial material into hardened soil, wherein the throwing amount of the mixed material is 0.2-0.4 kg per mu.
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