A biological soil conditioner containing mineral powder and preparation method thereof

Abstract

The present invention discloses a biological soil conditioner containing mineral powder and a preparation method thereof, and belongs to the technical field of microbial soil conditioners. The conditioner of the present invention comprises the following raw materials in parts by weight: 20-40 parts of composite microbial bacterial solution, 10-20 parts of mineral powder, and 15-20 parts of auxiliary material carrier. The present invention screens two highly acid-resistant growth-promoting bacterial fungal microorganisms, and loads them into highly active rice husk biochar after composite. The raw materials cooperate with each other, which can improve the aluminum pollution of acid-aluminum soil, improve soil fertility, promote plant growth, and enrich the soil microbial community structure, and has great application prospects.

Description

Technical Field

[0001] The invention belongs to the technical field of microbial soil conditioners, and in particular relates to a biological soil conditioner containing mineral powder and a preparation method thereof. Background Art

[0002] Soil acidification is one of the obstacles to soil, which seriously restricts the sustainable development of agriculture. Acidic soils include red soil, brick red soil, red soil, yellow soil and other soil types. After soil acidification, the weathering of primary and secondary minerals containing aluminum in the soil is accelerated to release a large amount of aluminum ions, forming aluminum compounds that can be absorbed by plants. Excessive absorption of aluminum by crops will not only reduce the quality of crop products, but also have a great impact on the growth of crops, especially the root system of crops, and even cause poisoning and death of crops. Soil acidification reduces the cation exchange capacity and base saturation, and the loss of soil mineral nutrients is serious; soil acidification also enhances the soil's fixation of phosphate and molybdate, and reduces the effectiveness of soil phosphorus and molybdenum.

[0003] In addition, long-term erosion of acidic soils has led to leaching of topsoil, loss of nutrients such as calcium, magnesium, and phosphorus, and aluminum toxicity in the soil, which has seriously inhibited the growth of crops. Therefore, it is necessary to improve acidic soils, reduce their exchangeable aluminum content, increase soil pH and exchangeable cation content, and ensure the safety of agricultural production in acidified soil areas.

[0004] At present, the existing technology for acidic soil treatment mainly adopts chemical improvement method, (1) applying lime to precipitate exchangeable aluminum into insoluble light-based aluminum, increase aluminum saturation to alleviate the inhibition of crop growth and increase soil pH. However, the application of lime has the following problems: in terms of application method, the usual application of lime on the soil surface cannot effectively neutralize the soil acidity of the central soil layer, and the large amount of lime used is also too costly. (2) Applying calcium magnesium phosphate fertilizer, calcium and magnesium are closely related to aluminum toxicity, and their alkalinity can neutralize the acidity of acid-aluminum soil, but excessive application can easily lead to resource waste and environmental pollution. (3) Applying organic matter to acidic soil, organic matter can effectively alleviate aluminum toxicity and improve phosphorus utilization, but the operation is cumbersome. The above three acidic soil conditioners can all have a certain improvement effect on acidic soil, increase soil pH, reduce the aluminum content in the soil, and thus improve soil acidity. However, all three conditioners have serious side effects, and the application frequency is frequent, the cost is too high, the operation is cumbersome, and it is easy to lead to resource waste and environmental pollution, and there is a serious risk of soil heavy metal pollution.

[0005] For example, Chinese patent application CN202111287964.5 discloses an acidic soil conditioner and a preparation method thereof. The acidic soil conditioner of the present invention comprises potassium-containing minerals, calcium-containing minerals, and zeolite. After the acidic soil conditioner of the present invention is applied to the soil, it can effectively increase the soil pH value, reduce the exchangeable aluminum and toxic aluminum content of the soil, and promote plant growth.

[0006] Biological improvement measures mainly include applying organic manure, crop rotation, using microbial agents, planting stress-resistant crops, etc. Compared with traditional chemical improvement methods for acidic soil, biological improvement methods have the advantages of high efficiency, easy availability, safety and cleanliness, and have been widely used in recent years. However, the current microbial soil conditioners have a single function, and most common microorganisms are not very active in acid-aluminum soils, so they cannot play a stable role. How to develop an efficient acidic soil conditioner is a technical problem that needs to be solved urgently. Summary of the invention

[0007] In view of the problems existing in the current prior art, the present invention selects two highly acid-resistant and growth-promoting bacterial and fungal microorganisms, composites them and loads them into highly active rice husk biochar, and simultaneously adds mineral powder. The raw materials cooperate with each other to achieve effective improvement of acid-aluminum soil and effective growth promotion of plants.

[0008] In order to achieve the above technical objectives, the technical solution adopted by the present invention is:

[0009] A biological soil conditioner containing mineral powder comprises the following raw materials in parts by weight: 20-40 parts of composite microbial liquid, 10-20 parts of mineral powder, and 15-20 parts of auxiliary material carrier.

[0010] Furthermore, the preparation method of the ore powder is: illite, potassium feldspar and calcium-containing ore powder are mixed according to a mass ratio of (5-10): (1-3): (3-6), and then ball milling is performed. After the ball milling is completed, the temperature is increased to 350-450°C at a heating rate of 0.5-1°C / min in a pure nitrogen atmosphere, and the temperature is kept at this temperature for 20-30 minutes; then the temperature is continued to be increased to 580-600°C, and the temperature is kept at this temperature for 30-40 minutes, and the ore powder is naturally cooled to room temperature, crushed, and ground evenly to obtain the ore powder.

[0011] Furthermore, the calcium-containing mineral powder is one or more of dolomite, limestone, and phosphate lime; the ball-to-material ratio of the ball milling treatment is 10-12:1, the ball milling speed is 100-200rpm, and the ball milling time is 40-60min.

[0012] Furthermore, the preparation method of the composite microbial liquid is as follows:

[0013] (1) Inoculate the strain of CGMCC No. 1.8860 of Forestomonas amyloliquefaciens in a liquid culture medium for liquid fermentation. The liquid fermentation conditions are as follows: temperature 25-28° C., rotation speed 170-200 rpm, fermentation time 1-2 days, obtain the fermentation liquid of Forestomonas amyloliquefaciens, dilute and apply to count the number of cells, and the cell concentration of the fermentation liquid of Forestomonas amyloliquefaciens is ≥ 1×10 8 cfu / mL;

[0014] (2) Inoculating the long-branched Trichoderma spores with the strain number CGMCC No. 5.1235 into a liquid culture medium for liquid fermentation, wherein the liquid culture medium is a PDA liquid culture medium, and the liquid fermentation conditions are: temperature 25-28° C., rotation speed 120-150 rpm, first fermentation under dark conditions for 2-3 days, and then fermentation under light conditions for 2-3 days, filtering to remove the hyphae, and obtaining the long-branched Trichoderma spore liquid, and determining the number of spores in the spore liquid by a hemocytometer. The spore concentration of the long-branched Trichoderma spore liquid is ≥1×10 8 Pieces / mL;

[0015] (3) centrifuging the fermentation broth of Forestomonas amyloliquefaciens, and resuspending the bacteria in sterile water with a volume ten times that of the fermentation broth to obtain a Forestomonas amyloliquefaciens broth;

[0016] (4) diluting the long-branch Trichoderma spore solution ten times with sterile water to obtain a long-branch Trichoderma solution;

[0017] (5) The amyloliquefaciens solution and the long-branched Trichoderma solution were mixed in a volume ratio of 1:1 to obtain a composite microbial solution.

[0018] Furthermore, step (1) Forestomonas amyloliquefaciens ( Silvimonas amylolytica ) is numbered CGMCC No1.8860, and its original deposit date is December 1, 2008. The strain was purchased from China General Microorganism Collection Center. The strain can be purchased through the collection center without repeated biological deposit. Step (1) The liquid culture medium is a modified LB medium, and its composition is: tryptone 10 g / L; yeast extract 5 g / L; sodium chloride 15 g / L; agar 19 g / L; pH adjusted to 4.5-5; sterilized at 121°C for 20 min.

[0019] Furthermore, in step (2) Trichoderma longifolia ( Trichoderma longibrachiatum) is numbered CGMCC No5.1235, and its original preservation date is September 8, 2006. The strain was purchased from China General Microorganism Collection Center. The strain can be purchased through the collection center without repeated biological preservation. Step (2) The composition of the PDA liquid culture medium is: 200 g / L potato, 20 g / L glucose, 15 g / L agar, and the pH is adjusted to 4.5-5; sterilize at 121°C for 20 min.

[0020] Furthermore, the auxiliary material carrier is a mixture of diatomaceous earth and rice husk biochar. The specific preparation method is: diatomaceous earth and rice husk are mixed in a mass ratio of 1: (10-15), crushed by a grinder and passed through a 2mm sieve, pyrolyzed at a heating rate of 2°C / min for 2h under anaerobic conditions, and the pyrolysis temperature is 400-600°C. After the pyrolysis is cooled to room temperature, it is ground through a 100-mesh sieve, washed with water, and dried in an oven at 80°C for 12h to prepare an auxiliary material carrier.

[0021] A method for preparing a biological soil conditioner containing mineral powder comprises the following preparation steps: after uniformly mixing an auxiliary material carrier and a microbial bacterial solution, transferring the mixture into a closed environment, evacuating the mixture to a vacuum degree of 0.05-0.06MPa, maintaining the vacuum for 1-2h, restoring the mixture to normal pressure, and standing the mixture; then adding mineral powder to the mixture, uniformly mixing the mixture, drying and granulating the mixture to obtain solid particles with a particle size of 2-5mm.

[0022] The soil conditioner of the present invention is used as a base fertilizer (base fertilizer). It is best to spread it evenly over the whole field before plowing. It can also be applied in furrows or holes. The amount per mu is applied according to the crop and soil conditions. Generally, when the soil pH value is 4-6, the amount is 20-30 kg / mu. The amount can be increased as appropriate for severely acidified soils.

[0023] Beneficial effects:

[0024] (1) The present invention selects two acid-resistant bacteria and fungi to achieve effective regulation of acid-aluminum soil; among them, the amyloliquefaciens strain CGMCC No. 1.8860 has multiple growth-promoting effects and the ability to produce organic acids such as citric acid. Citric acid can not only chelate aluminum ions (Al 3+), reducing its toxicity in the soil, and promoting plant growth by improving the structure of rhizosphere microbial communities; and the biological organic acids produced can stimulate the activity of other beneficial microorganisms to a certain extent. The long-branch Trichoderma fungus is compounded with Forest Mononas amyloliquefaciens. The long-branch Trichoderma can improve soil nutrients, increase soil fertility, and inhibit pathogens by secreting a variety of secondary metabolites, thereby indirectly promoting the growth of other beneficial microorganisms; at the same time, the long-branch Trichoderma fungus has a variety of growth-promoting effects, especially its phosphorus-dissolving effect, which can efficiently convert inorganic phosphorus into soluble phosphorus. Its phosphate group can combine with aluminum ions in the soil to form aluminum phosphate, effectively reducing the aluminum content in the soil; phosphorus-dissolving microorganisms can also improve the content of phosphorus in the soil, increase the available phosphorus in the soil for plants, and promote plant growth. The two beneficial microorganisms cooperate with each other and work synergistically to achieve effective improvement of acid-aluminum soil.

[0025] (2) The present invention also adds a mineral powder component consisting of illite, potassium feldspar and calcium-containing mineral powder, wherein illite has a high cation exchange capacity and can stabilize the pH value of the soil by adsorbing and fixing aluminum ions in the soil. Under acidic conditions, illite can form stable compounds with aluminum ions, thereby reducing the active aluminum content in the soil and improving the pH value of the soil. Potassium feldspar releases potassium in the soil, which can significantly increase the exchangeable potassium content in the soil and enhance the disease resistance and stress resistance of plants. The addition of calcium-containing mineral powder can improve the soil aggregation structure, increase the water retention and air permeability of the soil by neutralizing the soil acidity and releasing calcium ions. This helps to improve the fertility of the soil and the growth conditions of crops.

[0026] (3) The present invention uses diatomite and rice husk biochar as microbial carriers. A small amount of diatomite and rice husk biochar are combined for high-temperature pyrolysis. Both have high specific surface areas and porous structures, which can provide a large number of microbial attachment sites. This property helps to improve the survival rate and activity of microorganisms, thereby enhancing their effect on soil remediation.

[0027] (4) In summary, the microbial soil conditioner of the present invention can improve aluminum pollution in acid-aluminum soil, improve soil fertility, promote plant growth, enrich the soil microbial community structure, and has great application prospects. BRIEF DESCRIPTION OF THE DRAWINGS

[0028] Figure 1 This is a growth state diagram of the present invention's Forestomonas amyloliquefaciens and Trichoderma longibrachiatum in a high-acid culture medium;

[0029] Figure 2 This is a growth state diagram of the Forestobacterium amyloliquefaciens and Trichoderma longibrachiatum in a high-acid and high-aluminum culture medium of the present invention;

[0030] Figure 3This is a graph showing the changes in rice yield in different experimental groups. DETAILED DESCRIPTION

[0031] The technical solution of the present invention is further described below in conjunction with specific embodiments, but is not limited thereto.

[0032] Example 1

[0033] A biological soil conditioner containing mineral powder comprises the following raw materials in parts by weight: 40 parts of composite microbial liquid, 20 parts of mineral powder, and 15 parts of auxiliary material carrier.

[0034] The preparation method of the ore powder is as follows: illite, potassium feldspar and calcium-containing ore powder are mixed in a mass ratio of 10:1:6, and then ball milled; after the ball milling is completed, the temperature is increased to 450°C at a heating rate of 0.5°C / min in a pure nitrogen atmosphere, and the temperature is kept for 20 minutes; then the temperature is continued to be increased to 580-600°C, and the temperature is kept for 30 minutes, and the mixture is naturally cooled to room temperature, crushed, and ground evenly to obtain the ore powder.

[0035] The calcium-containing mineral powder is dolomite; the ball-to-material ratio of the ball milling treatment is 10:1, the ball milling speed is 100-200rpm, and the ball milling time is 40min.

[0036] The preparation method of the composite microbial liquid is as follows:

[0037] (1) Inoculate the strain CGMCC No. 1.8860 of Forestomonas amyloliquefaciens into a liquid culture medium for liquid fermentation. The liquid fermentation conditions are as follows: temperature 25-28° C., rotation speed 170-200 rpm, fermentation time 1 day, obtain the fermentation liquid of Forestomonas amyloliquefaciens, dilute and apply to count the number of cells, and the cell concentration of the fermentation liquid of Forestomonas amyloliquefaciens is ≥ 1×10 8 cfu / mL;

[0038] (2) Inoculating the long-branched Trichoderma spores with the strain number CGMCC No. 5.1235 into a liquid culture medium for liquid fermentation, wherein the liquid culture medium is a PDA liquid culture medium, and the liquid fermentation conditions are: temperature 25-28° C., rotation speed 120-150 rpm, first fermentation under dark conditions for 2 days, and then fermentation under light conditions for 2 days, filtering to remove the hyphae, and obtaining the long-branched Trichoderma spore liquid, and determining the number of spores in the spore liquid by a hemocytometer. The spore concentration of the long-branched Trichoderma spore liquid is ≥1×10 8 Pieces / mL;

[0039] (3) centrifuging the fermentation broth of Forestomonas amyloliquefaciens, and resuspending the bacteria in sterile water with a volume ten times that of the fermentation broth to obtain a Forestomonas amyloliquefaciens broth;

[0040] (4) diluting the long-branch Trichoderma spore solution ten times with sterile water to obtain a long-branch Trichoderma solution;

[0041] (5) The amyloliquefaciens solution and the long-branched Trichoderma solution were mixed in a volume ratio of 1:1 to obtain a composite microbial solution.

[0042] Step (1) The strain number of Silvimonas amylolytica is CGMCCNo1.8860, and the original preservation time is December 1, 2008. The strain was purchased from the China General Microorganism Collection Center. The strain can be purchased through the collection center without repeated biological preservation. Step (1) The liquid culture medium is a modified LB medium, and its composition is: tryptone 10g / L; yeast extract 5g / L; sodium chloride 15g / L; agar 19g / L; pH adjusted to 4.5-5; sterilized at 121℃ for 20min.

[0043] The strain number of the long-branched Trichoderma in step (2) is CGMCC No. 5.1235, and the original preservation time is September 8, 2006. The strain was purchased from the China General Microorganism Collection Center. The strain can be purchased through the collection center without repeated biological preservation. The composition of the PDA liquid culture medium in step (2) is: 200 g / L potato, 20 g / L glucose, 15 g / L agar, and the pH is adjusted to 4.5-5; sterilized at 121° C. for 20 min.

[0044] The auxiliary material carrier is a mixture of diatomaceous earth and rice husk biochar. The specific preparation method is: diatomaceous earth and rice husk are mixed in a mass ratio of 1: (10), crushed by a grinder and passed through a 2 mm sieve, pyrolyzed at a heating rate of 2°C / min for 2 hours under anaerobic conditions, and the pyrolysis temperature is 600°C. After the pyrolysis is cooled to room temperature, it is ground through a 100 mesh sieve, washed with water, and dried in an oven at 80°C for 12 hours to prepare an auxiliary material carrier.

[0045] A method for preparing a biological soil conditioner containing mineral powder comprises the following preparation steps: after uniformly mixing an auxiliary material carrier and a microbial bacterial solution, transferring the mixture into a closed environment, evacuating the mixture to a vacuum degree of 0.05-0.06MPa, maintaining the vacuum for 1 hour, restoring the mixture to normal pressure, and standing the mixture; then adding mineral powder to the mixture, uniformly mixing the mixture, drying and granulating the mixture to obtain solid particles with a particle size of 2-5mm.

[0046] The soil conditioner in this embodiment is used as a base fertilizer (bottom fertilizer). It is best to spread it evenly over the whole field before plowing. It can also be applied in furrows or holes. The amount per mu is applied according to the crop and soil conditions. Generally, when the soil pH value is 4-6, the amount is 20-30 kg / mu. For severely acidified soils, the amount can be increased as appropriate.

[0047] Example 2

[0048] A biological soil conditioner containing mineral powder comprises the following raw materials in parts by weight: 30 parts of composite microbial liquid, 15 parts of mineral powder, and 20 parts of auxiliary material carrier.

[0049] The preparation method of the ore powder is as follows: illite, potassium feldspar and calcium-containing ore powder are mixed in a mass ratio of 5:3:3, and then ball milled; after the ball milling is completed, the temperature is increased to 400°C at a heating rate of 0.5-1°C / min in a pure nitrogen atmosphere, and the temperature is kept for 30 minutes; then the temperature is continued to be increased to 580-600°C, and the temperature is kept for 40 minutes, and the mixture is naturally cooled to room temperature, crushed, and ground evenly to obtain the ore powder.

[0050] The calcium-containing mineral powder is limestone; the ball-to-material ratio of the ball milling treatment is 12:1, the ball milling speed is 100-200rpm, and the ball milling time is 60min.

[0051] The preparation method of the composite microbial liquid is as follows:

[0052] (1) Inoculate the strain of CGMCC No. 1.8860 of Forestomonas amyloliquefaciens in a liquid culture medium for liquid fermentation. The liquid fermentation conditions are as follows: temperature 25-28° C., rotation speed 170-200 rpm, fermentation time 2 days, obtain the fermentation liquid of Forestomonas amyloliquefaciens, dilute and apply to count the number of cells, and the cell concentration of the fermentation liquid of Forestomonas amyloliquefaciens is ≥ 1×10 8 cfu / mL;

[0053] (2) Inoculating the spores of Trichoderma longibrachiatum with a strain number of CGMCC No. 5.1235 into a liquid culture medium for liquid fermentation, wherein the liquid culture medium is a PDA liquid culture medium, and the liquid fermentation conditions are: temperature 25-28° C., rotation speed 120-150 rpm, first fermentation under dark conditions for 2 days, and then fermentation under light conditions for 3 days, filtering to remove the hyphae, and obtaining a Trichoderma longibrachiatum spore liquid, and determining the number of spores in the spore liquid by a hemocytometer. The spore concentration of the Trichoderma longibrachiatum spore liquid is ≥1×10 8 Pieces / mL;

[0054] (3) centrifuging the fermentation broth of Forestomonas amyloliquefaciens, and resuspending the bacteria in sterile water with a volume ten times that of the fermentation broth to obtain a Forestomonas amyloliquefaciens broth;

[0055] (4) diluting the long-branch Trichoderma spore solution ten times with sterile water to obtain a long-branch Trichoderma solution;

[0056] (5) The amyloliquefaciens solution and the long-branched Trichoderma solution were mixed in a volume ratio of 1:1 to obtain a composite microbial solution.

[0057] The Silvimonas amylolytica in step (1) is the same as in Example 1; the liquid culture medium in step (1) is a modified LB culture medium, and its composition is the same as in Example 1.

[0058] The long-branched Trichoderma in step (2) is the same as that in Example 1; the composition of the PDA liquid culture medium in step (2) is the same as that in Example 1.

[0059] The auxiliary material carrier is a mixture of diatomaceous earth and rice husk biochar. The specific preparation method is: diatomaceous earth and rice husk are mixed in a mass ratio of 1:15, crushed by a grinder and passed through a 2mm sieve, pyrolyzed at a heating rate of 2°C / min for 2h under anaerobic conditions, and the pyrolysis temperature is 500°C. After the pyrolysis is cooled to room temperature, it is ground through a 100-mesh sieve, washed with water, and dried in an oven at 80°C for 12h to prepare an auxiliary material carrier.

[0060] A method for preparing a biological soil conditioner containing mineral powder comprises the following preparation steps: after uniformly mixing an auxiliary material carrier and a microbial bacterial solution, transferring the mixture into a closed environment, evacuating the mixture to a vacuum degree of 0.05-0.06MPa, maintaining the vacuum for 2 hours, restoring the mixture to normal pressure, and standing the mixture; then adding mineral powder to the mixture, uniformly mixing the mixture, drying and granulating the mixture to obtain solid particles with a particle size of 2-5mm.

[0061] Example 3

[0062] A biological soil conditioner containing mineral powder comprises the following raw materials in parts by weight: 20 parts of composite microbial liquid, 10 parts of mineral powder, and 15 parts of auxiliary material carrier.

[0063] The preparation method of the ore powder is as follows: illite, potassium feldspar and calcium-containing ore powder are mixed in a mass ratio of 10:2:5, and then ball milled; after the ball milling is completed, the temperature is increased to 350°C at a heating rate of 0.5-1°C / min in a pure nitrogen atmosphere, and the temperature is kept for 30 minutes; then the temperature is continued to be increased to 580-600°C, and the temperature is kept for 40 minutes, and the mixture is naturally cooled to room temperature, crushed, and ground evenly to obtain the ore powder.

[0064] The calcium-containing mineral powder is phosphate lime; the ball-to-material ratio of the ball milling treatment is 12:1, the ball milling speed is 100-200rpm, and the ball milling time is 60min.

[0065] The preparation method of the composite microbial liquid is as follows:

[0066] (1) Inoculate the strain of CGMCC No. 1.8860 of Forestomonas amyloliquefaciens in a liquid culture medium for liquid fermentation. The liquid fermentation conditions are as follows: temperature 25-28° C., rotation speed 170-200 rpm, fermentation time 2 days, obtain the fermentation liquid of Forestomonas amyloliquefaciens, dilute and apply to count the number of cells, and the cell concentration of the fermentation liquid of Forestomonas amyloliquefaciens is ≥ 1×10 8 cfu / mL;

[0067] (2) Inoculating the spores of Trichoderma longibrachiatum with a strain number of CGMCC No. 5.1235 into a liquid culture medium for liquid fermentation, wherein the liquid culture medium is a PDA liquid culture medium, and the liquid fermentation conditions are: temperature 25-28° C., rotation speed 120-150 rpm, first fermentation under dark conditions for 3 days, and then fermentation under light conditions for 3 days, filtering to remove the hyphae, and obtaining a Trichoderma longibrachiatum spore liquid, and determining the number of spores in the spore liquid by a hemocytometer. The spore concentration of the Trichoderma longibrachiatum spore liquid is ≥1×10 8 Pieces / mL;

[0068] (3) centrifuging the fermentation broth of Forestomonas amyloliquefaciens, and resuspending the bacteria in sterile water with a volume ten times that of the fermentation broth to obtain a Forestomonas amyloliquefaciens broth;

[0069] (4) diluting the long-branch Trichoderma spore solution ten times with sterile water to obtain a long-branch Trichoderma solution;

[0070] (5) The amyloliquefaciens solution and the long-branched Trichoderma solution were mixed in a volume ratio of 1:1 to obtain a composite microbial solution.

[0071] The Silvimonas amylolytica in step (1) is the same as in Example 1; the liquid culture medium in step (1) is a modified LB culture medium, and its composition is the same as in Example 1.

[0072] The long-branched Trichoderma in step (2) is the same as that in Example 1; the composition of the PDA liquid culture medium in step (2) is the same as that in Example 1.

[0073] The auxiliary material carrier is a mixture of diatomaceous earth and rice husk biochar. The specific preparation method is: diatomaceous earth and rice husk are mixed in a mass ratio of 1:13, crushed by a grinder and passed through a 2mm sieve, pyrolyzed at a heating rate of 2°C / min for 2h under anaerobic conditions, and the pyrolysis temperature is 400°C. After the pyrolysis is cooled to room temperature, it is ground through a 100-mesh sieve, washed with water, and dried in an oven at 80°C for 12h to prepare an auxiliary material carrier.

[0074] A method for preparing a biological soil conditioner containing mineral powder comprises the following preparation steps: after uniformly mixing an auxiliary material carrier and a microbial bacterial solution, transferring the mixture into a closed environment, evacuating the mixture to a vacuum degree of 0.05-0.06MPa, maintaining the vacuum for 2 hours, restoring the mixture to normal pressure, and standing the mixture; then adding mineral powder to the mixture, uniformly mixing the mixture, drying and granulating the mixture to obtain solid particles with a particle size of 2-5mm.

[0075] Comparative Example 1

[0076] In this comparative example, except that only the strain number of Forestomonas amyloliquefaciens CGMCC No. 1.8860 was used in the preparation of the composite microbial liquid, the other raw materials and preparation methods were the same as those in Example 3. That is:

[0077] A biological soil conditioner containing mineral powder comprises the following raw materials in parts by weight: 20 parts of microbial liquid, 10 parts of mineral powder, and 15 parts of auxiliary material carrier.

[0078] The preparation method of the microbial liquid is as follows:

[0079] (1) Inoculate the strain of CGMCC No. 1.8860 of Forestomonas amyloliquefaciens in a liquid culture medium for liquid fermentation. The liquid fermentation conditions are as follows: temperature 25-28° C., rotation speed 170-200 rpm, fermentation time 2 days, obtain the fermentation liquid of Forestomonas amyloliquefaciens, dilute and apply to count the number of cells, and the cell concentration of the fermentation liquid of Forestomonas amyloliquefaciens is ≥ 1×10 8 cfu / mL;

[0080] (2) Centrifuging the fermentation broth of Forestomonas amyloliquefaciens, resuspending the bacteria in sterile water with a volume ten times that of the fermentation broth, to obtain a Forestomonas amyloliquefaciens solution, i.e., a microbial bacterial solution.

[0081] Comparative Example 2

[0082] In this comparative example, except that only the long-branched Trichoderma with the strain number CGMCC No. 5.1235 was used in the preparation of the composite microbial liquid, the other raw materials and preparation methods were the same as those in Example 3. That is:

[0083] A biological soil conditioner containing mineral powder comprises the following raw materials in parts by weight: 20 parts of microbial liquid, 10 parts of mineral powder, and 15 parts of auxiliary material carrier.

[0084] The preparation method of the microbial liquid is as follows:

[0085] (1) Inoculating the spores of Trichoderma longibrachiatum with a strain number of CGMCC No. 5.1235 into a liquid culture medium for liquid fermentation, wherein the liquid culture medium is a PDA liquid culture medium, and the liquid fermentation conditions are: temperature 25-28° C., rotation speed 120-150 rpm, first fermentation under dark conditions for 3 days, and then fermentation under light conditions for 3 days, filtering to remove hyphae, and obtaining a Trichoderma longibrachiatum spore liquid, and determining the number of spores in the spore liquid by a hemocytometer. The spore concentration of the Trichoderma longibrachiatum spore liquid is ≥1×10 8 Pieces / mL;

[0086] (2) diluting the long-branch Trichoderma spore liquid ten times with sterile water to obtain a long-branch Trichoderma liquid, which is a composite microbial liquid.

[0087] Comparative Example 3

[0088] In this comparative example, except that the volume ratio of Forestobacillus amyloliquefaciens and Trichoderma longibrachiatum was changed in the preparation of the composite microbial liquid, the other raw materials and preparation method were the same as those in Example 3. That is:

[0089] A biological soil conditioner containing mineral powder comprises the following raw materials in parts by weight: 20 parts of composite microbial liquid, 10 parts of mineral powder, and 15 parts of auxiliary material carrier.

[0090] The preparation method of the composite microbial liquid is as follows:

[0091] (1) Inoculate the strain of CGMCC No. 1.8860 of Forestomonas amyloliquefaciens in a liquid culture medium for liquid fermentation. The liquid fermentation conditions are as follows: temperature 25-28° C., rotation speed 170-200 rpm, fermentation time 2 days, obtain the fermentation liquid of Forestomonas amyloliquefaciens, dilute and apply to count the number of cells, and the cell concentration of the fermentation liquid of Forestomonas amyloliquefaciens is ≥ 1×10 8 cfu / mL;

[0092] (2) Inoculating the spores of Trichoderma longibrachiatum with a strain number of CGMCC No. 5.1235 into a liquid culture medium for liquid fermentation, wherein the liquid culture medium is a PDA liquid culture medium, and the liquid fermentation conditions are: temperature 25-28° C., rotation speed 120-150 rpm, first fermentation under dark conditions for 3 days, and then fermentation under light conditions for 3 days, filtering to remove the hyphae, and obtaining a Trichoderma longibrachiatum spore liquid, and determining the number of spores in the spore liquid by a hemocytometer. The spore concentration of the Trichoderma longibrachiatum spore liquid is ≥1×10 8 Pieces / mL;

[0093] (3) centrifuging the fermentation broth of Forestomonas amyloliquefaciens, and resuspending the bacteria in sterile water with a volume ten times that of the fermentation broth to obtain a Forestomonas amyloliquefaciens broth;

[0094] (4) diluting the long-branch Trichoderma spore solution ten times with sterile water to obtain a long-branch Trichoderma solution;

[0095] (5) The amyloliquefaciens solution and the long-branched Trichoderma solution were mixed in a volume ratio of 2:1 to obtain a composite microbial solution.

[0096] Comparative Example 4

[0097] In this comparative example, except that the volume ratio of Forestobacillus amyloliquefaciens and Trichoderma longibrachiatum was changed in the preparation of the composite microbial liquid, the other raw materials and preparation method were the same as those in Example 3. That is:

[0098] A biological soil conditioner containing mineral powder comprises the following raw materials in parts by weight: 20 parts of composite microbial liquid, 10 parts of mineral powder, and 15 parts of auxiliary material carrier.

[0099] The preparation method of the composite microbial liquid is as follows:

[0100] (1) Inoculate the strain of CGMCC No. 1.8860 of Forestomonas amyloliquefaciens in a liquid culture medium for liquid fermentation. The liquid fermentation conditions are as follows: temperature 25-28° C., rotation speed 170-200 rpm, fermentation time 2 days, obtain the fermentation liquid of Forestomonas amyloliquefaciens, dilute and apply to count the number of cells, and the cell concentration of the fermentation liquid of Forestomonas amyloliquefaciens is ≥ 1×10 8 cfu / mL;

[0101] (2) Inoculating the spores of Trichoderma longibrachiatum with a strain number of CGMCC No. 5.1235 into a liquid culture medium for liquid fermentation, wherein the liquid culture medium is a PDA liquid culture medium, and the liquid fermentation conditions are: temperature 25-28° C., rotation speed 120-150 rpm, first fermentation under dark conditions for 3 days, and then fermentation under light conditions for 3 days, filtering to remove the hyphae, and obtaining a Trichoderma longibrachiatum spore liquid, and determining the number of spores in the spore liquid by a hemocytometer. The spore concentration of the Trichoderma longibrachiatum spore liquid is ≥1×10 8 Pieces / mL;

[0102] (3) centrifuging the fermentation broth of Forestomonas amyloliquefaciens, and resuspending the bacteria in sterile water with a volume ten times that of the fermentation broth to obtain a Forestomonas amyloliquefaciens broth;

[0103] (4) diluting the long-branch Trichoderma spore solution ten times with sterile water to obtain a long-branch Trichoderma solution;

[0104] (5) The amyloliquefaciens solution and the long-branched Trichoderma solution were mixed in a volume ratio of 1:2 to obtain a composite microbial solution.

[0105] Comparative Example 5

[0106] In this comparative example, except that the type of Trichoderma longifolia was changed in the preparation of the composite microbial liquid, the rest of the raw materials and preparation method were the same as those in Example 3. That is:

[0107] The preparation method of the composite microbial liquid is as follows:

[0108] (1) Inoculate the strain of CGMCC No. 1.8860 of Forestomonas amyloliquefaciens in a liquid culture medium for liquid fermentation. The liquid fermentation conditions are as follows: temperature 25-28° C., rotation speed 170-200 rpm, fermentation time 2 days, obtain the fermentation liquid of Forestomonas amyloliquefaciens, dilute and apply to count the number of cells, and the cell concentration of the fermentation liquid of Forestomonas amyloliquefaciens is ≥ 1×10 8 cfu / mL;

[0109] (1) Inoculating the spores of Trichoderma longibrachiatum with a strain number of CGMCC No. 3.6906 into a liquid culture medium for liquid fermentation, wherein the liquid culture medium is a PDA liquid culture medium, and the liquid fermentation conditions are: temperature 25-28° C., rotation speed 120-150 rpm, and the number of spores in the spore liquid is determined by a hemocytometer, and the spore concentration of the Trichoderma longibrachiatum spore liquid is ≥1×10 8 When the number of cells / mL is reached, the culture is completed;

[0110] (2) centrifuging the fermentation broth of Forestomonas amyloliquefaciens, and resuspending the bacteria in sterile water with a volume ten times that of the fermentation broth to obtain a Forestomonas amyloliquefaciens broth;

[0111] (3) diluting the long-branch Trichoderma spore solution ten times with sterile water to obtain a long-branch Trichoderma solution;

[0112] (4) The amyloliquefaciens solution and the long-branched Trichoderma solution were mixed in a volume ratio of 1:1 to obtain a composite microbial solution.

[0113] Step (2) The long-branched Trichoderma with strain number CGMCC No. 3.6906 was purchased from China General Microorganism Collection Center and the original preservation date was May 24, 2005.

[0114] Comparative Example 6

[0115] In this comparative example, except for changing the type of fungi in the preparation of the composite microbial liquid, the rest of the raw materials and preparation methods are the same as those in Example 3. That is, Aspergillus niger with similar phosphate-dissolving and growth-promoting functions is selected to replace Trichoderma longibrachiatum to prepare the composite liquid:

[0116] The preparation method of the composite microbial liquid is as follows:

[0117] (1) Inoculate the strain of CGMCC No. 1.8860 of Forestomonas amyloliquefaciens in a liquid culture medium for liquid fermentation. The liquid fermentation conditions are as follows: temperature 25-28° C., rotation speed 170-200 rpm, fermentation time 2 days, obtain the fermentation liquid of Forestomonas amyloliquefaciens, dilute and apply to count the number of cells, and the cell concentration of the fermentation liquid of Forestomonas amyloliquefaciens is ≥ 1×10 8 cfu / mL;

[0118] (2) Inoculating Aspergillus niger spores with a strain number of CCTCC HF 2008599 into a liquid culture medium for liquid fermentation, wherein the liquid culture medium is a PDA liquid culture medium, and the liquid fermentation conditions are: a temperature of 25-28° C., a rotation speed of 120-150 rpm, and determining the number of spores in the spore liquid by a hemocytometer counting method, and culturing until the spore concentration of the Aspergillus niger spore liquid is ≥1×10 8 When the number of cells / mL is reached, the culture is completed;

[0119] (3) centrifuging the fermentation broth of Forestomonas amyloliquefaciens, and resuspending the bacteria in sterile water with a volume ten times that of the fermentation broth to obtain a Forestomonas amyloliquefaciens broth;

[0120] (4) diluting the Aspergillus niger spore solution ten-fold with sterile water to obtain a Trichoderma longifolia solution;

[0121] (5) The Forestomonas amyloliquefaciens solution and the Aspergillus niger solution were mixed in a volume ratio of 1:1 to obtain a composite microbial solution.

[0122] Step (2) Aspergillus niger with strain number CCTCC HF 2008599 was purchased from China Center for Type Culture Collection, and the original preservation date was July 2, 2005.

[0123] Performance Testing

[0124] Test of strains' acid and aluminum resistance

[0125] Seed solution preparation: The strain number of CGMCC No. 1.8860 was inoculated into LB liquid medium for cultivation at a bacterial concentration of 1×10 7 cfu / mL, and the seed solution of Forestomonas amyloliquefaciens was obtained.

[0126] The long-branched Trichoderma strain numbered CGMCC No. 5.1235 was transferred to the LB plate and cultured in a 25°C incubator for 2 days. When the spores cover the entire plate, it can be used to prepare the spore suspension. 5 mL of LB medium was taken to rinse the spores, and 2 mL of spore suspension was accurately taken to 100 mL of LB liquid medium. It was cultured in a shaking incubator at 25°C and 180 r / min for 24 hours as the seed liquid. The spore concentration was determined to be 1×10 7Pieces / mL.

[0127] Determination of acid resistance: The seed solution to be tested was inoculated into LB liquid medium with a pH of 4 at a 1% inoculation rate, and cultured at 25-28°C and 180 r / min for 2 days. The OD 600 , an equal amount of sterile water was used as a control, and the experiment was repeated three times.

[0128] Determination of aluminum tolerance: Prepare LB liquid medium containing aluminum concentration of 5 mM (pH 3), and inoculate the seed solution into the medium at an inoculum rate of 1%, culture at 25-28℃ and 180 r / min for 2 days, and measure the OD 600 , an equal amount of sterile water was used as a control, and the experiment was repeated three times.

[0129] Table 1 OD of strains cultured in LB liquid medium with pH 4 for 2 days 600

[0130]

[0131] From the data in Table 1 and Figure 1 We can see that the two strains screened by the present invention can grow normally in an environment of pH 4 and have a certain acid resistance.

[0132] Table 2 OD of strains cultured in LB liquid medium with aluminum concentration of 5 mM (pH 3) for 2 days 600

[0133]

[0134] From the data in Table 2 and Figure 2 We can see that the two strains screened by the present invention can grow normally in an environment with an aluminum concentration of 5 mM and have certain acid and aluminum resistance.

[0135] Test of strain growth-promoting ability:

[0136] Determination of phosphate solubilization ability: The molybdenum antimony colorimetric method was used to determine the phosphate solubilization ability of the strain. Seven 50 mL Nessler colorimetric tubes were selected, and 0, 0.5, 1, 3, 5, 10 and 15 mL of phosphate standard solution were added respectively, and the volume was made up to 50 mL with sterile water. 1 mL of molybdate solution and 10% ascorbic acid solution were added to the Nessler test tubes. Mix well and let stand for 30 minutes. The phosphorus content was 0 μg mL -1 The solution was zero-adjusted and its OD was measured. 700 The strains to be tested were inoculated into 100 mL of inorganic phosphorus liquid culture medium at a 1% (V / V) inoculation rate. Each strain was repeated 3 times. The non-inoculated culture medium was used as a control. The culture medium was kept at 25-28°C and 180 r·min.-1 After 7 days of cultivation, the supernatant was taken to determine the phosphorus content.

[0137] Determine different types and concentrations of plant hormone standards and draw a standard curve. Inoculate the seed solution at 1% (V / V) in LB liquid medium and rotate at 25-28℃ and 180 r·min -1 The culture was cultured at a constant temperature for 3 days under the same conditions. The pH of the bacterial solution was adjusted to 4 with hydrochloric acid solution and allowed to stand for 30 min. After centrifugation, the supernatant was mixed with ethyl acetate and shaken for 25 min, allowed to stand for 20 min, and the organic components were collected. The organic solvent was removed by drying at 40 °C using a rotary evaporator, methanol was added to dissolve the dry matter, and filtered through a 0.22 μm filter membrane. The types and contents of plant hormones produced by the strain were determined by high performance liquid chromatography.

[0138] Determination of organic acid production by strains:

[0139] The citric acid standard was diluted to an appropriate concentration, and its peak time and peak area were determined. The chromatographic column was Thermo Hypersil BDS-C18, 5 μm; methanol and 0.02 mol·L -1 NaH 2 PO (4 140 mL:860 mL) was used as the mobile phase; the running time for each sample was set to 25 min, and the flow rate was 1 mL min -1 . The column temperature was 30 °C and the detection wavelength was 210 nm. The supernatant of the fermentation broth of the strain to be tested was collected, filtered through a 0.22 μm water filter membrane, and then determined according to the method of Gao Wei et al. (Gao Wei, Zuo Zhenyu, Li Lingling, et al. Screening and identification of a highly efficient phosphate-solubilizing bacterium and its phosphate-solubilizing performance [J]. Bulletin of Microbiology, 2022, 49(9): 3873-3889.).

[0140] Table 3 Test of growth-promoting ability of different strains

[0141]

[0142] Soil improvement and planting experiments

[0143] The experimental field is planted with rice all year round. Before the experiment, the field pH was 4.50, organic matter was 51.3 g / kg, alkaline nitrogen was 98 mg / kg, available phosphorus was 12.6 mg / kg, available potassium was 105 mg / kg, and cation exchange capacity was 1.22 mol / kg.

[0144] The comparative test of different treatment modes is designed as follows:

[0145] CK: conventional fertilization N, P 2 O 5 and K 2O dosage was 120, 60 and 150 kg / hm 2 ;

[0146] S1: conventional fertilization (same as CK) + soil conditioner of Example 1 20 kg / mu;

[0147] S2: conventional fertilization (same as CK) + soil conditioner of Example 2 20 kg / mu;

[0148] S3: conventional fertilization (same as CK) + soil conditioner of Example 3 20 kg / mu;

[0149] S4: conventional fertilization (same as CK) + soil conditioner of comparative example 1 20kg / mu;

[0150] S5: conventional fertilization (same as CK) + soil conditioner of comparative example 2 20kg / mu;

[0151] S6: conventional fertilization (same as CK) + soil conditioner of comparative example 3 20kg / mu;

[0152] S7: conventional fertilization (same as CK) + soil conditioner of comparative example 4 20kg / mu;

[0153] S8: conventional fertilization (same as CK) + soil conditioner of comparative example 5 20kg / mu;

[0154] S9: conventional fertilization (same as CK) + soil conditioner of comparative example 6 20kg / mu;

[0155] Each treatment was repeated 3 times, and all experimental results were averaged.

[0156] The plots were arranged randomly, with protective rows set up around the plots. The ridges between the plots were separated by film covering. Each plot was independently irrigated and drained, and field management measures such as field irrigation and pest control were the same as those in the field. The types of nitrogen fertilizer, phosphorus fertilizer and potassium fertilizer were urea, calcium magnesium phosphate fertilizer and potassium chloride, respectively. All phosphorus fertilizer was applied as base fertilizer, and 50% of nitrogen fertilizer and potassium fertilizer were applied as base fertilizer and 50% as topdressing.

[0157] Sample collection and analysis:

[0158] At rice harvest, soil samples from 0 to 20 cm were collected from each plot using a soil drill. After air-drying and grinding, soil pH was determined by potentiometric method (water-soil ratio 2.5:1), organic matter was determined by potassium dichromate oxidation-volumetric method, alkaline hydrolysis nitrogen was determined by alkaline diffusion method, available phosphorus was determined by sodium bicarbonate extraction-molybdenum antimony-scandium colorimetry, available potassium was determined by ammonium acetate extraction-flame photometry, and exchangeable acid (exchangeable aluminum) was determined by potassium chloride extraction-titration method according to the "Technical Specifications for Soil Analysis".

[0159] Table 4 Changes in soil acidity indexes in different experimental groups

[0160]

[0161] Table 5 Changes in Soil Fertility Indexes of Different Experimental Groups

[0162]

[0163] From Tables 4 - 5 and Figure 3 the data, it can be seen that the soil conditioner of the embodiment of the present invention can effectively reduce the soil acidity value and exchangeable aluminum, and optimize and improve the soil fertility level. However, in Comparative Examples 1 - 6 where the core microbial composition was changed, and the strain composition and dosage were changed, the synergistic balance effect of the functional strains was destroyed, thus resulting in a decline in the soil effect and the rice yield of the planting effect.

[0164] It should be noted that the above - mentioned embodiments are only some embodiments of the preferred ways to implement the present invention, rather than all embodiments. Obviously, based on the above - mentioned embodiments of the present invention, all other embodiments obtained by those of ordinary skill in the art without creative efforts shall fall within the protection scope of the present invention.

Claims

1. A biological soil conditioner containing mineral powder, characterized in that: The invention comprises the following raw materials in parts by weight: 20-40 parts of composite microbial liquid, 10-20 parts of ore powder, and 15-20 parts of auxiliary material carrier; the preparation method of the composite microbial liquid is as follows: (1) Inoculate the strain of CGMCC No. 1.8860 of Forestomonas amyloliquefaciens in a liquid culture medium for liquid fermentation. The liquid fermentation conditions are as follows: temperature 25-28° C., rotation speed 170-200 rpm, fermentation time 1-2 days, obtain the fermentation liquid of Forestomonas amyloliquefaciens, dilute and apply to count the number of cells, and the cell concentration of the fermentation liquid of Forestomonas amyloliquefaciens is ≥1×10 8 cfu / mL; (2) Inoculating the long-branched Trichoderma spores with the strain number CGMCC No. 5.1235 into a liquid culture medium for liquid fermentation, wherein the liquid culture medium is a PDA liquid culture medium, and the liquid fermentation conditions are: temperature 25-28° C., rotation speed 120-150 rpm, first fermentation under dark conditions for 2-3 days, and then fermentation under light conditions for 2-3 days, filtering to remove the hyphae, and obtaining the long-branched Trichoderma spore liquid, and determining the number of spores in the spore liquid by a hemocytometer method, wherein the spore concentration of the long-branched Trichoderma spore liquid is ≥1×10 8 Pieces / mL; (3) centrifuging the fermentation broth of Forestomonas amyloliquefaciens, and resuspending the bacteria in sterile water with a volume ten times that of the fermentation broth to obtain a Forestomonas amyloliquefaciens broth; (4) diluting the long-branch Trichoderma spore solution ten times with sterile water to obtain a long-branch Trichoderma solution; (5) mixing the Forestomonas amylovora solution and the Trichoderma longibrachiatum solution in a volume ratio of 1:1 to obtain a composite microbial solution; The preparation method of the ore powder comprises: mixing illite, potassium feldspar and calcium-containing ore powder according to a mass ratio of (5-10):(1-3):(3-6), and then subjecting the mixture to ball milling. After the ball milling, heating the mixture to 350-450° C. at a heating rate of 0.5-1° C. / min in a pure nitrogen atmosphere, and heat-keeping the mixture for 20-30 min; then heating the mixture to 580-600° C., heat-keeping the mixture for 30-40 min, naturally cooling the mixture to room temperature, crushing the mixture, and grinding the mixture uniformly to obtain the ore powder. The calcium-containing mineral powder is one or more of dolomite, limestone, and phosphate lime; the ball-to-material ratio of ball milling treatment is (10-12):1, the ball milling speed is 100-200rpm, and the ball milling is 40-60min; the auxiliary material carrier is a mixture of diatomaceous earth and rice husk biochar, and the specific preparation method is: diatomaceous earth and rice husk are mixed in a mass ratio of 1: (10-15), crushed by a pulverizer and passed through a 2mm sieve, pyrolyzed at a heating rate of 2°C / min for 2h under anaerobic conditions, and the pyrolysis temperature is 400-600°C. After the pyrolysis is cooled to room temperature, it is ground through a 100-mesh sieve, washed with water, and dried in an oven at 80°C for 12h to prepare an auxiliary material carrier.

2. A method for preparing the biological soil conditioner containing mineral powder according to claim 1, characterized in that: The preparation method comprises the following steps: after the auxiliary material carrier and the microbial liquid are mixed evenly, they are transferred into a closed environment, evacuated to a vacuum degree of 0.05-0.06MPa, the vacuum is maintained for 1-2 hours, and then the pressure is restored to normal pressure and allowed to stand; ore powder is then added to the mixture, the mixture is mixed evenly, and then dried and granulated to obtain solid particles with a particle size of 2-5mm.

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