Bio-organic fertilizer fermentation agent, preparation method and application thereof

Abstract

The application discloses a kind of bio-organic fertilizer fermentation microbial inoculum and its preparation method and application, belong to microbial technical field.The microbial inoculum includes active microorganism and synergistic component, and the mass ratio of two is 1: (0.3-0.6).The application significantly improves fermentation efficiency and product quality by optimizing active microorganism composition and synergistic component.The microbial inoculum can not only quickly decompose lignocellulose structure in straw, but also effectively reduce nutrient loss, improve the maturity and fertilizer efficiency stability of organic fertilizer.By scientifically compounding aspergillus fungal spore powder and cellulose-degrading bacteria active bacteria preparation, and combining specific synergistic components, the microbial inoculum exhibits excellent synergistic effect during composting process.This technical scheme not only solves the problems of slow fermentation, incomplete composting and other problems in traditional organic fertilizer production, but also provides a new solution for the resource utilization of agricultural waste.

Description

TECHNICAL FIELD

[0001] The present application belongs to the field of microbial technology, and particularly relates to a bio-organic fertilizer fermentation agent and a preparation method thereof. BACKGROUND

[0002] With the development of modern agriculture, the use of chemical fertilizers is increasing, far exceeding the application amount required by crops and the unreasonable fertilization method brings problems such as low fertilizer utilization rate, soil degradation and environmental pollution.

[0003] Organic fertilizer is a kind of fertilizer with rich nutrients, stable fertilizer supply and soil improvement function, which is produced by technical processing of organic waste. Organic fertilizer can significantly improve the environmental pollution problems caused by excessive use of chemical fertilizer, and effectively improve the yield and quality of crops. However, the raw material cost of traditional commercial organic fertilizer is high, and some chemical substances may be introduced in the manufacturing process, which may have potential impact on the soil ecological environment.

[0004] With the rapid development of agriculture and economy in China, the unreasonable disposal of a large amount of crop straw and livestock manure has brought great pressure on the environment. Composting of agricultural waste to produce organic fertilizer is an effective means of resource utilization, and is also an important way to improve economic efficiency. However, composting treatment has the problems of slow natural fermentation, serious nutrient loss and incomplete composting due to the presence of a large amount of cellulose, lignin and other difficult-to-degrade substances in the compost material. The use of compound microbial inoculants to promote the degradation of lignocellulose in the compost and improve the fermentation efficiency is the key to solving the problem.

[0005] For example, Chinese patent application CN201710676656.9 discloses an organic fertilizer fermentation agent, which is prepared by mixing bacterial preparation, mold preparation, yeast preparation and actinomycete preparation in the following weight ratio: bacterial preparation:mold preparation:yeast preparation:actinomycete preparation = 2-7:2-8:2:1-3. However, the existing microbial agents have the disadvantages of single strain, poor activity and high cost, especially for straw, cow dung, sheep dung and other wastes with high cellulose content, the fermentation effect is poor. Especially for straw waste, the lignocellulose structure of straw is the biggest obstacle. The physical barrier formed by lignin wraps cellulose and hemicellulose, making it difficult for microorganisms to contact the substrate. Although physical crushing (such as steam explosion) can destroy the structure, the equipment investment is large, and small and medium-sized enterprises are difficult to bear. Biological pretreatment relies on cellulase, but the cost of enzyme preparation accounts for more than 30%, and the enzyme activity is easily inhibited. This seriously restricts the resource utilization efficiency of agricultural waste and the development of organic fertilizer industry.

[0006] Therefore, it is an urgent technical problem to develop a fermentation agent with high efficiency of promoting decomposition and nutrient loss, and to produce high-quality organic fertilizer with comprehensive nutrients and long-term fertilizer supply. SUMMARY

[0007] The application provides a bio-organic fertilizer fermentation agent, which is characterized by optimizing the proportion of active microorganisms and synergistic components, thereby significantly improving fermentation efficiency and product quality. The agent can not only quickly decompose lignocellulose structure in straw, but also effectively reduce nutrient loss, improve composting degree and fertilizer efficiency stability of organic fertilizer. In addition, the application also provides a specific preparation method of the agent, which ensures that the agent has the characteristics of simple operation and low cost in actual application. The agent exhibits excellent synergistic effect in the composting process by scientific compounding of Aspergillus spore powder and cellulose-degrading bacteria active bacteria preparation and combining with specific synergistic components. This technical scheme not only solves the problems of slow fermentation and incomplete composting in traditional organic fertilizer production, but also provides a new solution for resource utilization of agricultural waste.

[0008] To achieve the above technical purposes, the technical scheme adopted by the application is:

[0009] A bio-organic fertilizer fermentation agent, which comprises active microorganisms and synergistic components, and the mass ratio of the two is 1: (0.3-0.6).

[0010] Further, the active microorganisms comprise Aspergillus spore powder and cellulose-degrading bacteria active bacteria preparation, and the mass ratio of the two is 1:1.

[0011] Further, the preparation method of the Aspergillus spore powder is:

[0012] (1) inoculate the Aspergillus ruber (Aspergillus ruber) with strain number CGMCC No 3.15548 into an activated culture medium A, and incubate at 25±2 DEG C for 3-5 days to obtain activated slant bacteria; Monascus purpureus

[0013] (2) inoculate the activated bacteria into a liquid seed culture medium, and culture at a shaking speed of 220-250 rpm and 23-25 DEG C for 48-72 hours, so that the concentration of bacteria in the bacterial solution is greater than or equal to 10 8 CFU / mL, to obtain a seed solution;

[0014] (3) inoculate the seed solution into a solid spore-producing substrate at an inoculation amount of 5%-8%, and then load the culture bag, and incubate at 23-25 DEG C and relative humidity of 75%-85% in the dark for 7-10 days, so that the surface of the substrate is covered with a red spore layer, and the spore density is greater than or equal to 10 9 individuals / g under microscopic observation;

[0015] ​(4) Add sterile water containing 0.05% Tween-80 to the sporulation substrate, stir to elute the spores, and filter through a 300-mesh filter cloth to remove mycelium residues; centrifuge the filtrate at 4°C and 3000 rpm for 10 minutes to collect the spore precipitate, and use vacuum freeze-drying treatment to obtain Aspergillus spore powder with a moisture content of ≤8% and a spore survival rate of ≥90%.

[0016] The strain of Aspergillus puniceus with the strain number of CGMCC No 3.15548 Monascus purpureus was purchased from the China General Microbiological Culture Collection Center, and the original preservation time was February 19, 2016. The strain can be purchased through the market channel.

[0017] Step (1): The composition of the activation medium A is: potato 200 g / L, glucose 20 g / L, peptone 5 g / L, agar 20 g / L, and the pH is natural.

[0018] Step (2): The composition of the liquid seed culture medium is: corn syrup dry powder 6-8 g / L, glucose 15 g / L, yeast extract 3 g / L, MgSO4·7H2O 0.5 g / L, and the pH is 6.0-6.5.

[0019] Step (3): The composition of the solid sporulation substrate is: bran 70%, corn cob powder 20%, soybean meal 5%, CaCO3 3%, KH2PO4 2%, and water is added to adjust the moisture content to 60%-65%.

[0020] Further, the preparation method of the cellulose-degrading bacteria live bacteria preparation is:

[0021] (1) Strain screening and activation: inoculate the Guangxi high-temperature actinomycete with the strain number of CGMCC No 4.7156 into the activation medium B, and place it in a constant temperature incubator at 42±2°C, avoiding light for 48-72 hours to obtain gray-white radial colonies;

[0022] (2) Seed liquid expansion culture: inoculate the activated colonies into the liquid culture medium with an inoculation amount of 6%-8%, and place it in a 40°C, 220 rpm shaker for 36-48 hours to obtain a bacterial liquid with a live bacteria concentration of ≥10 9 CFU / mL;

[0023] (3) Solid fermentation for bacteria production: inoculate the seed liquid into the fermentation substrate with an inoculation amount of 10%-12%, put it into a gas-permeable fermentation bag, and place it in an environment with a temperature of 45±2°C and a relative humidity of 70%-80% for 5-7 days. The fermentation end point characteristics are: the substrate is brownish yellow and loose, the mycelium density is ≥10 5 strands / g under a microscope, and the live bacteria number is ≥10 10 CFU / g. After drying and crushing, the cellulose-degrading bacteria live bacteria preparation is obtained.

[0024] Actinomyces wuyangensis (CGMCC No 4.7156) Thermoactinomyces guangxiensis It was purchased from China General Microbiological Culture Collection Center, and the original preservation time was January 22, 2014. The strain can be purchased through the market channel.

[0025] Further, the composition of the activated culture medium B in step (1) is: soluble starch 20 g / L, KNO3 1 g / L, K2HPO4 0.5 g / L, MgSO4·7H2O 0.5 g / L, NaCl 0.5 g / L, FeSO4·7H2O 0.01 g / L, pH 7.2-7.4.

[0026] Further, the composition of the liquid culture medium in step (2) is: on the basis of removing agar in the activated culture medium B, adding 5 g / L sodium carboxymethyl cellulose.

[0027] Further, the composition of the fermentation substrate in step (3) is: straw powder (crushed to ≤2 mm) 50%, bran 30%, soybean meal 15%, CaCO3 5%, and water is added to adjust the moisture content to 60%-65%, and sterilized at 121℃ for 30 minutes.

[0028] Further, the synergistic component includes the following mass parts of raw materials: grass ash 30-40 parts, bone meal 5-10 parts, brown sugar 10-15 parts.

[0029] The raw materials used in the application are commercially available.

[0030] A preparation method of a bio-organic fertilizer fermentation agent, comprising the following steps:

[0031] (1) preparing Aspergillus fungal spore powder and cellulose-degrading bacteria active bacteria preparation, and mixing the two uniformly according to a mass ratio of 1:1;

[0032] (2) mixing the active microorganisms with the synergistic component in proportion, first screening the grass ash and bone meal in the synergistic component to 80-100 meshes to ensure uniform particle size; then dissolving the brown sugar and an appropriate amount of water in the stirring equipment to form a sugar solution; uniformly spraying the sugar solution on the surface of the grass ash and bone meal mixture, adding the active microorganisms while stirring, controlling the stirring speed to be 60-80 rpm, and the time to be 5-10 minutes, so that the components are fully mixed; after mixing, the material is placed in an environment with a temperature of 25-30℃ and a relative humidity of 50%-60% for 1-2 hours to promote the adaptation of the microorganisms to the environment and preliminary activation; finally, the mixed agent is packed in a sealed packaging bag, and vacuum packaging technology is used to avoid external pollution.

[0033] The bio-organic fertilizer fermentation agent of the application is used for straw fermentation to prepare organic fertilizer.

[0034] The use of the microbial agent of the present application is as follows: the crop straw is crushed into small pieces of 3-6 cm long, urea is added to adjust to about 25:1, the microbial agent is added to water and dispersed uniformly, and then sprayed uniformly on the material pile with a water sprayer, finally the pile is turned over while spraying clean water, the moisture content is adjusted to about 60%, the mass ratio of the microbial agent and the straw is 50g:10kg. Fermentation is carried out for 15-20 days under natural conditions, the pile is turned over every 3-4 days, and after fermentation is completed, the straw bio-organic fertilizer is obtained by drying to a moisture content of not more than 30%.

[0035] Beneficial effects:

[0036] (1) Two functional strains Guangxi high-temperature actinomycete and purple red koji mold capable of degrading cellulose efficiently are screened, wherein the purple red koji mold can secrete high-activity laccase efficiently under the micro-acid environment in the early stage of fermentation, effectively disassembling the lignin structure of the straw, and the Guangxi high-temperature actinomycete can secrete high-activity heat-resistant cellulase to degrade the exposed cellulose rapidly, and the two strains can produce significant synergistic effect after mixed in equal proportion, and the cellulose degradation rate is greatly improved by the combination of the two strains.

[0037] (2) The synergist composed of wood ash, bone meal and brown sugar is added, the wood ash in the synergist can neutralize the organic acid generated in the early stage of composting, maintain the suitable pH value, and thus improve the activity of laccase secreted by the purple red koji mold; the bone meal as a long-acting phosphorus source can not only promote the growth and reproduction of microorganisms, but also supplement the mineral elements lost in the fermentation process; the brown sugar as a quick-acting carbon source can rapidly activate the metabolic activity of microorganisms and enhance the initial colonization ability of the microbial agent. By scientific proportioning of the components, the microbial agent exhibits excellent fermentation performance in composting treatment, greatly shortens the fermentation period, and improves the composting degree and efficiency of the organic fertilizer. In addition, the production process of the microbial agent is simple, the raw material cost is low, and the large-scale production is easy to realize, which has significant economic and environmental benefits. BRIEF DESCRIPTION OF DRAWINGS

[0038] Figure 1 It is a schematic diagram of strain antagonism experiment of the present application;

[0039] Figure 2 It is a photo of antagonism experiment results of the purple red koji mold and Guangxi high-temperature actinomycete, wherein A is Guangxi high-temperature actinomycete, and B is purple red koji mold;

[0040] Figure 3 It is a photo of transparent circle of the purple red koji mold and Guangxi high-temperature actinomycete on Congo red medium;

[0041] Figure 4 It is a scanning electron microscope graph of the surface of rice straw before and after composting degradation. DETAILED DESCRIPTION

[0042] The technical solutions of the present application are further described below in combination with specific embodiments, but are not limited thereto.

[0043] Embodiment 1

[0044] A bio-organic fertilizer fermentation agent, the agent comprising active microorganisms and synergistic components, both in a mass ratio of 1:0.3.

[0045] The active microorganisms comprise Aspergillus spore powder and cellulose-degrading bacteria active bacteria preparation, both in a mass ratio of 1:1.

[0046] The preparation method of the Aspergillus spore powder is as follows:

[0047] (1) inoculate Aspergillus ruber with a strain number of CGMCC No 3.15548 into an activated culture medium A, and incubate at a constant temperature of 25±2℃ for 3-5 days to obtain activated slant bacteria; Monascus purpureus

[0048] (2) inoculate the activated bacteria into a liquid seed culture medium, and culture at a shaking speed of 220-250 rpm and a temperature of 23-25℃ for 48-72 hours until the concentration of bacteria in the bacterial solution is ≥10 8 CFU / mL, to obtain a seed solution;

[0049] (3) inoculate the seed solution into a solid spore-producing substrate at an inoculation amount of 5%-8%, and then incubate in a transparent culture bag in a dark environment at a temperature of 23-25℃ and a relative humidity of 75%-85% for 7-10 days until the surface of the substrate is covered with a red spore layer and the spore density is ≥10 9 individuals / g under microscopic observation;

[0050] (4) add sterile water containing 0.05% Tween-80 to the spore-producing substrate, stir to elute the spores, and filter the mycelium residues through a 300-mesh filter cloth; centrifuge the filtrate at 3000 rpm for 10 minutes at 4℃ to collect the spore precipitate, and use vacuum freeze-drying treatment to obtain Aspergillus spore powder with a water content of ≤8% and a spore survival rate of ≥90%.

[0051] Aspergillus ruber with a strain number of CGMCC No 3.15548 Monascus purpureus is purchased from the China General Microbiological Culture Collection Center, and the original preservation time is February 19, 2016. The strain can be purchased through the market channel.

[0052] Step (1) The composition of the activated culture medium A is as follows: potato 200g / L, glucose 20g / L, peptone 5g / L, and agar 20g / L, with a natural pH.

[0053] ​Step (2) the composition of the liquid seed culture medium: corn syrup dry powder 6-8 g / L, glucose 15 g / L, yeast extract 3 g / L, MgSO4·7H2O 0.5 g / L, pH 6.0-6.5.

[0054] Step (3) the composition of the solid spore-producing substrate: bran 70%, corn cob powder 20%, soybean meal 5%, CaCO3 3%, KH2PO4 2%, add water to 60%-65% moisture content.

[0055] The preparation method of the cellulose-degrading bacteria live bacteria preparation is:

[0056] (1) strain screening and activation: inoculate Guangxi high-temperature actinomycetes with strain number CGMCC No 4.7156 into activation culture medium B, and place it in a constant temperature incubator at 42±2°C, avoiding light for 48-72 hours to obtain gray-white radial colonies;

[0057] (2) seed liquid expansion culture: inoculate the activated colonies into a liquid culture medium with an inoculation amount of 6%-8%, and place it in a 40°C, 220 rpm shaker for 36-48 hours, with a bacteria concentration of ≥10 9 CFU / mL at the end of the culture;

[0058] (3) solid fermentation to produce bacteria: inoculate the seed liquid into the fermentation substrate at an inoculation amount of 10%-12%, put it into a gas-permeable fermentation bag, and place it in an environment with a temperature of 45±2°C and a relative humidity of 70%-80% for 5-7 days. The fermentation end point characteristics are as follows: the substrate is brownish yellow and loose, the mycelium density is ≥10 5 g / m, the viable bacteria number is ≥10 10 CFU / g, after drying and crushing, the cellulose-degrading bacteria live bacteria preparation is obtained.

[0059] Guangxi high-temperature actinomycetes with strain number CGMCC No 4.7156 Thermoactinomyces guangxiensis ) is purchased from the China General Microbiological Culture Collection Center, and the original preservation time is January 22, 2014. The strain can be purchased through the market channel.

[0060] Step (1) the composition of the activation culture medium B: soluble starch 20 g / L, KNO3 1 g / L, K2HPO4 0.5 g / L, MgSO4·7H2O 0.5 g / L, NaCl 0.5 g / L, FeSO4·7H2O 0.01 g / L, pH 7.2-7.4.

[0061] Step (2) the composition of the liquid culture medium is to remove agar from the activation culture medium B and add 5 g / L sodium carboxymethyl cellulose.

[0062] The fermentation substrate of step (3) is composed of straw powder (crushed to ≤2 mm) 50%, bran 30%, soybean meal 15%, CaCO3 5%, and water is added to adjust the moisture content to 60%-65%, sterilized at 121 ℃ for 30 minutes.

[0063] The synergistic component comprises the following mass parts of raw materials: wood ash 30 parts, bone meal 5 parts, brown sugar 10 parts.

[0064] Each raw material used in this embodiment is commercially available.

[0065] A preparation method of a bio-organic fertilizer fermentation agent, comprising the following steps:

[0066] (1) Prepare Aspergillus spore powder and cellulose-degrading bacteria active bacteria preparation, and mix the two according to a mass ratio of 1:1;

[0067] (2) Mix the active microorganisms with the synergistic component in proportion, first sieve the wood ash and bone meal in the synergistic component to 80-100 mesh to ensure uniform particle size; then add brown sugar and an appropriate amount of water to dissolve in the stirring equipment to form a sugar solution; uniformly spray the sugar solution on the surface of the wood ash and bone meal mixture, add the active microorganisms while stirring, control the stirring speed to be 60-80 rpm, and the time to be 5-10 minutes, so that the components are fully mixed; after mixing, place the material in an environment with a temperature of 25-30 ℃ and a relative humidity of 50%-60% for 1-2 hours to promote the adaptation of the microorganisms to the environment and preliminary activation; finally, pack the mixed agent into a sealed packaging bag using vacuum packaging technology to avoid external pollution.

[0068] Example 2

[0069] A bio-organic fertilizer fermentation agent, the agent comprises active microorganisms and a synergistic component, and the mass ratio of the two is 1:0.6.

[0070] The active microorganisms comprise Aspergillus spore powder and cellulose-degrading bacteria active bacteria preparation, and the mass ratio of the two is 1:1.

[0071] The preparation method of the Aspergillus spore powder is:

[0072] (1) Inoculate the Monascus purpureus strain numbered CGMCC No 3.15548 into activated culture medium A and incubate at a constant temperature of 25±2 ℃ for 3-5 days to obtain activated slant bacteria; Monascus purpureus

[0073] (2) Inoculate the activated bacteria into a liquid seed culture medium, and culture at a shaking speed of 220-250 rpm and a temperature of 23-25 ℃ for 48-72 hours, so that the concentration of bacteria in the bacterial solution is ≥10 8 CFU / mL, to obtain a seed solution; ​

[0074] (3) Inoculate the seed culture at a rate of 5%-8% into the solid sporulation substrate, place it in a breathable culture bag, and culture it in the dark at 23-25℃ and 75%-85% relative humidity for 7-10 days until a red spore layer covers the substrate surface. Simultaneously, observe the spore density under a microscope to ensure it is ≥10⁻⁶. 9 pcs / g;

[0075] (4) Add sterile water containing 0.05% Tween-80 to the sporulation substrate, stir to wash away the spores, filter through a 300-mesh filter cloth to remove mycelial residues; collect the spore precipitate by centrifuging the filtrate at 4℃ and 3000rpm for 10 minutes, and freeze-dry it under vacuum to obtain Aspergillus spore powder with a moisture content ≤8% and a spore survival rate ≥90%.

[0076] Aspergillus violaceus strain numbered CGMCC No. 3.15548 Monascus purpureus This strain was purchased from the China General Microbiological Culture Collection Center, with an original deposit date of February 19, 2016. It is available for purchase through commercial channels.

[0077] Step (1) The composition of activation medium A is: 200g / L potato, 20g / L glucose, 5g / L peptone, 20g / L agar, pH natural.

[0078] Step (2) The liquid seed culture medium consists of: 6-8 g / L corn steep liquor powder, 15 g / L glucose, 3 g / L yeast extract, 0.5 g / L MgSO4・7H2O, and pH 6.0-6.5.

[0079] Step (3) The solid sporulation substrate consists of: 70% wheat bran, 20% corn cob powder, 5% soybean meal, 3% CaCO3, 2% KH2PO4, and water is added to adjust the moisture content to 60%-65%.

[0080] The preparation method of the cellulose-degrading bacteria live bacterial preparation is as follows:

[0081] (1) Strain screening and activation: Guangxi high-temperature actinomycetes with strain number CGMCC No4.7156 were inoculated into activation medium B and placed in a constant temperature incubator at 42±2℃ for 48-72 hours in the dark to obtain grayish-white radial colonies;

[0082] (2) Seed culture expansion: Inoculate activated colonies into liquid culture medium at an inoculation rate of 6%-8%, and incubate at 40℃ and 220rpm for 36-48 hours. The viable bacterial concentration in the culture solution should be ≥10%. 9 CFU / mL terminates culture;

[0083] (3) solid fermentation production bacteria: inoculate the seed liquid into the fermentation substrate at an inoculation amount of 10%-12%, load into a gas permeable fermentation bag, and place in an environment of 45±2 DEG C and relative humidity of 70%-80% for 5-7 days. The fermentation end point characteristics are: the substrate is brownish yellow and loose, the mycelium density is greater than or equal to 10 5 CFU / g, the viable bacterial count is greater than or equal to 10 10 CFU / g after drying and crushing, to obtain a cellulose-degrading bacteria viable bacterial preparation.

[0084] Guangxi actinomycete (Thermoactinomyces sp.) with strain number CGMCC No 4.7156 Thermoactinomyces guangxiensis ) purchased from the China General Microbiological Culture Collection Center, and the original preservation time is January 22, 2014. The strain can be purchased through a commercial channel.

[0085] Step (1) The composition of the activated culture medium B is: soluble starch 20 g / L, KNO3 1 g / L, K2HPO4 0.5 g / L, MgSO4·7H2O 0.5 g / L, NaCl 0.5 g / L, FeSO4·7H2O 0.01 g / L, and pH 7.2-7.4.

[0086] Step (2) The composition of the liquid culture medium is: remove the agar from the activated culture medium B and add 5 g / L sodium carboxymethyl cellulose.

[0087] Step (3) The composition of the fermentation substrate is: straw powder (crushed to less than or equal to 2 mm) 50%, bran 30%, soybean meal 15%, and CaCO3 5%, and water is added to adjust the moisture content to 60%-65%, and sterilized at 121 DEG C for 30 minutes.

[0088] The synergistic component comprises the following mass parts of raw materials: grass ash 40 parts, bone meal 10 parts, and brown sugar 15 parts.

[0089] A preparation method of a bio-organic fertilizer fermentation bacterial agent, comprising the following steps:

[0090] (1) Prepare Aspergillus fungal spore powder and cellulose-degrading bacteria viable bacterial preparation, and mix the two uniformly according to a mass ratio of 1:1;

[0091] (2) Mixing the active microorganism with the synergistic component in proportion, first sieving the wood ash and bone meal in the synergistic component to 80-100 mesh to ensure uniform particle size; then dissolving the brown sugar and an appropriate amount of water in a stirring device to form a sugar solution; uniformly spraying the sugar solution on the surface of the wood ash and bone meal mixture, adding the active microorganism while stirring, controlling the stirring speed at 60-80 rpm, and the time at 5-10 minutes, so that the components are fully mixed; after mixing, the material is placed in an environment with a temperature of 25-30°C and a relative humidity of 50%-60% for 1-2 hours to promote the adaptation of the microorganism to the environment and preliminary activation; finally, the mixed inoculant is packed in sealed packaging bags using vacuum packaging technology to avoid external pollution.

[0092] Comparative Example 1

[0093] In this comparative example, except that only a single microorganism Guangxi thermophilic actinomycete is used in the active microorganism, the remaining raw materials and process steps are the same as in Example 1. That is:

[0094] A bio-organic fertilizer fermentation inoculant, the inoculant comprising active microorganisms and a synergistic component, the mass ratio of the two being 1:0.3.

[0095] The active microorganism is a cellulose-degrading bacteria live bacteria preparation.

[0096] The preparation method of the cellulose-degrading bacteria live bacteria preparation is:

[0097] (1) Strain selection and activation: inoculate the Guangxi thermophilic actinomycete with strain number CGMCC No 4.7156 into the activation medium B, and place it in a constant temperature incubator at 42±2°C, avoiding light for 48-72 hours to obtain gray-white radial colonies;

[0098] (2) Seed liquid expansion culture: inoculate the activated colonies into the liquid medium with an inoculation amount of 6%-8%, and place it in a 40°C, 220rpm shaking bed for 36-48 hours, with a bacteria concentration of ≥10 9 CFU / mL at the end of the culture;

[0099] (3) Solid fermentation to produce bacteria: inoculate the seed liquid into the fermentation substrate at an inoculation amount of 10%-12%, put it into a breathable fermentation bag, and place it in an environment with a temperature of 45±2°C and a relative humidity of 70%-80% for 5-7 days. The fermentation endpoint characteristics are: the substrate is brownish yellow and loose, the mycelium density is ≥10 5 strands / g, and the viable bacteria count is ≥10 10 CFU / g. After drying, it is crushed to obtain a cellulose-degrading bacteria live bacteria preparation.

[0100] Guangxi thermophilic actinomycete with strain number CGMCC No 4.7156 Thermoactinomyces guangxiensisIt was purchased from China General Microbiological Culture Collection Center, and the original preservation time was January 22, 2014. The strain can be purchased through the market channel.

[0101] The composition of step (1) activated culture medium B is: soluble starch 20 g / L, KNO3 1 g / L, K2HPO4 0.5 g / L, MgSO4·7H2O 0.5 g / L, NaCl 0.5 g / L, FeSO4·7H2O 0.01 g / L, pH 7.2-7.4.

[0102] The composition of step (2) liquid culture medium is to remove agar on the basis of activated culture medium B, and add 5 g / L sodium carboxymethyl cellulose.

[0103] The composition of step (3) fermentation substrate is: straw powder (crushed to ≤2 mm) 50%, bran 30%, soybean meal 15%, CaCO3 5%, add water to 60%-65% moisture content, 121℃ sterilization for 30 minutes.

[0104] The synergistic component includes the following mass parts of raw materials: wood ash 30 parts, bone meal 5 parts, brown sugar 10 parts.

[0105] A preparation method of a bio-organic fertilizer fermentation agent, comprising the following steps:

[0106] (1) preparing a cellulose-degrading bacteria live bacteria preparation;

[0107] (2) mixing the active microorganisms with the synergistic components in proportion, first screening the wood ash and bone meal in the synergistic components to 80-100 mesh to ensure uniform particle size; then adding brown sugar and a proper amount of water to dissolve in the stirring equipment to form a sugar solution; uniformly spraying the sugar solution on the surface of the wood ash and bone meal mixture, adding active microorganisms while stirring, controlling the stirring speed to 60-80 rpm, and the time to 5-10 minutes, so that the components are fully mixed; after mixing, the material is placed in an environment with a temperature of 25-30℃ and a relative humidity of 50%-60% for 1-2 hours to promote the adaptation of the microorganisms to the environment and preliminary activation; finally, the mixed agent is packed in a sealed packaging bag, and vacuum packaging technology is used to avoid external pollution.

[0108] Comparative Example 2

[0109] In this comparative example, except that only a single microorganism Monascus purpureus is used in the active microorganisms, the remaining raw materials and process steps are the same as in Example 1. That is:

[0110] A bio-organic fertilizer fermentation agent, the agent contains active microorganisms and synergistic components, and the mass ratio of the two is 1:0.3.

[0111] The active microorganism is Aspergillus spore powder.

[0112] The preparation method of the Aspergillus spore powder is as follows:

[0113] (1) inoculate the Aspergillus ruber with strain number CGMCC No 3.15548 into an activated culture medium A, and incubate at a constant temperature of 25±2℃ for 3-5 days to obtain an activated slant strain; Monascus purpureus

[0114] (2) inoculate the activated strain into a liquid seed culture medium, and cultivate at a shaking speed of 220-250 rpm and a temperature of 23-25℃ for 48-72 hours until the concentration of the bacterial bodies in the bacterial liquid is ≥10 8 CFU / mL to obtain a seed liquid;

[0115] (3) inoculate the seed liquid into a solid spore-producing medium at an inoculation amount of 5%-8%, and then load the medium into a breathable culture bag, and cultivate at a temperature of 23-25℃ and a relative humidity of 75%-85% in the dark for 7-10 days until the surface of the medium is covered with a red spore layer and the spore density is ≥10 9 individuals / g under microscopic observation;

[0116] (4) add sterile water containing 0.05% Tween-80 into the spore-producing medium, stir to elute the spores, and remove the mycelium residues through 300-mesh filter cloth; centrifuge the filtrate at 3000 rpm for 10 minutes at 4℃ to collect the spore precipitate, and then treat the spore precipitate by vacuum freeze-drying to obtain Aspergillus spore powder with a water content of ≤8% and a spore survival rate of ≥90%.

[0117] The Aspergillus ruber with strain number CGMCC No 3.15548 Monascus purpureus is purchased from the China General Microbiological Culture Collection Center, and the original preservation time is February 19, 2016. The strain can be purchased through the market channel.

[0118] The composition of the activated culture medium A in step (1) is as follows: potato 200 g / L, glucose 20 g / L, peptone 5 g / L, and agar 20 g / L, with a natural pH.

[0119] The composition of the liquid seed culture medium in step (2) is as follows: corn syrup dry powder 6-8 g / L, glucose 15 g / L, yeast extract 3 g / L, and MgSO4·7H2O 0.5 g / L, with a pH of 6.0-6.5.

[0120] The composition of the solid spore-producing medium in step (3) is as follows: bran 70%, corn cob powder 20%, soybean meal 5%, CaCO3 3%, and KH2PO4 2%, with water added to a water content of 60%-65%.

[0121] The synergistic component comprises the following raw materials by mass: wood ash 30 parts, bone meal 5 parts, and brown sugar 10 parts. ​

[0122] A preparation method of a bio-organic fertilizer fermentation agent, comprising the following steps:

[0123] (1) preparing Aspergillus spore powder;

[0124] (2) mixing the active microorganism and the synergistic component in proportion, first sieving the wood ash and bone meal in the synergistic component to 80-100 meshes to ensure uniform particle size; then dissolving brown sugar and an appropriate amount of water in a stirring device to form a sugar solution; uniformly spraying the sugar solution on the surface of the wood ash and bone meal mixture, adding the active microorganism while stirring, controlling the stirring speed to be 60-80 rpm, and the time to be 5-10 minutes, so that the components are fully mixed; after mixing is completed, the material is placed in an environment with a temperature of 25-30°C and a relative humidity of 50%-60% for 1-2 hours to promote the adaptation of the microorganism to the environment and preliminary activation; finally, the mixed agent is packed into a sealed packaging bag using vacuum packaging technology to avoid external pollution.

[0125] Comparative Example 3

[0126] In this comparative example, the mass ratio of Aspergillus spore powder and cellulose-degrading bacteria active bacteria preparation in the active microorganism is changed to 2:1, and the remaining raw materials and process steps are the same as in Example 1. That is:

[0127] A bio-organic fertilizer fermentation agent, comprising active microorganisms and synergistic components, with a mass ratio of 1:3.

[0128] The active microorganism comprises Aspergillus spore powder and cellulose-degrading bacteria active bacteria preparation, with a mass ratio of 2:1.

[0129] A preparation method of a bio-organic fertilizer fermentation agent, comprising the following steps:

[0130] (1) preparing Aspergillus spore powder and cellulose-degrading bacteria active bacteria preparation, and mixing the two in a mass ratio of 2:1;

[0131] (2) mixing the active microorganism and the synergistic component in proportion, first sieving the wood ash and bone meal in the synergistic component to 80-100 meshes to ensure uniform particle size; then dissolving brown sugar and an appropriate amount of water in a stirring device to form a sugar solution; uniformly spraying the sugar solution on the surface of the wood ash and bone meal mixture, adding the active microorganism while stirring, controlling the stirring speed to be 60-80 rpm, and the time to be 5-10 minutes, so that the components are fully mixed; after mixing is completed, the material is placed in an environment with a temperature of 25-30°C and a relative humidity of 50%-60% for 1-2 hours to promote the adaptation of the microorganism to the environment and preliminary activation; finally, the mixed agent is packed into a sealed packaging bag using vacuum packaging technology to avoid external pollution.

[0132] Comparative Example 4

[0133] The comparative example, except for changing the mass ratio of active microorganism Aspergillus spore powder, cellulose-degrading bacteria live bacteria preparation to 1:2, the rest of the raw materials and process steps are the same as example 1. That is:

[0134] A bio-organic fertilizer fermentation agent, the agent contains active microorganisms and synergistic components, the mass ratio of the two is 1:3.

[0135] The active microorganism contains Aspergillus spore powder and cellulose-degrading bacteria live bacteria preparation, the mass ratio of the two is 1:2.

[0136] A preparation method of a bio-organic fertilizer fermentation agent, comprising the following steps:

[0137] (1) Prepare Aspergillus spore powder and cellulose-degrading bacteria live bacteria preparation, and mix the two according to a mass ratio of 1:2;

[0138] (2) Mix the active microorganism with the synergistic component according to the proportion, first sieve the wood ash and bone meal in the synergistic component to 80-100 mesh to ensure uniform particle size; then dissolve the brown sugar in the stirring equipment with an appropriate amount of water to form a sugar solution; evenly spray the sugar solution on the surface of the mixture of wood ash and bone meal, add the active microorganism while stirring, control the stirring speed to be 60-80 rpm, and the time to be 5-10 minutes, so that the components are fully mixed; after mixing, the material is placed in an environment with a temperature of 25-30°C and a relative humidity of 50%-60% for 1-2 hours to promote the adaptation of the microorganism to the environment and preliminary activation; finally, the mixed agent is packed in a sealed bag and vacuum packaged to avoid external pollution.

[0139] Comparative example 5

[0140] The comparative example, except for changing the strain type for preparing Aspergillus spore powder, the rest of the raw materials and process steps are the same as example 1. That is:

[0141] A bio-organic fertilizer fermentation agent, the agent contains active microorganisms and synergistic components, the mass ratio of the two is 1:0.3.

[0142] The active microorganism contains Aspergillus spore powder and cellulose-degrading bacteria live bacteria preparation, the mass ratio of the two is 1:1.

[0143] The preparation method of the Aspergillus spore powder is:

[0144] (1) Inoculate the Monascus purpureus strain numbered CGMCC No 3.15543 into the activated culture medium A and incubate at a constant temperature of 25±2°C for 3-5 days to obtain the activated slant strain; Monascus purpureus

[0145] ​(2) Inoculate the activated bacterial strain into liquid seed culture medium and incubate for 48-72 hours at a shaking speed of 220-250 rpm and a temperature of 23-25°C, until the bacterial concentration in the culture is ≥10%. 8 CFU / mL was used to obtain the seed culture;

[0146] (3) Inoculate the seed culture at a rate of 5%-8% into the solid sporulation substrate, place it in a breathable culture bag, and culture it in the dark at 23-25℃ and 75%-85% relative humidity for 7-10 days until a red spore layer covers the substrate surface. Simultaneously, observe the spore density under a microscope to ensure it is ≥10⁻⁶. 9 pcs / g;

[0147] (4) Add sterile water containing 0.05% Tween-80 to the sporulation substrate, stir to wash away the spores, filter through a 300-mesh filter cloth to remove mycelial residues; collect the spore precipitate by centrifuging the filtrate at 4℃ and 3000rpm for 10 minutes, and freeze-dry it under vacuum to obtain Aspergillus spore powder with a moisture content ≤8% and a spore survival rate ≥90%.

[0148] Aspergillus violaceus strain numbered CGMCC No. 3.15543 Monascus purpureus This strain was purchased from the China General Microbiological Culture Collection Center, with an original deposit date of February 19, 2016. It is available for purchase through commercial channels.

[0149] Comparative Example 6

[0150] In this comparative example, except for changing the strain type of thermotolerant actinomycetes in the cellulose-degrading bacteria live bacterial preparation, the raw materials and process steps are the same as in Example 1. That is:

[0151] The cultivation method is the same as in Example 1.

[0152] The preparation method of the cellulose-degrading bacteria live bacterial preparation is as follows:

[0153] (1) Strain screening and activation: The thermophilic actinomycetes with strain number GDMCC No4.160 were inoculated into activation medium B and placed in a constant temperature incubator at 42±2℃ for 48-72 hours in the dark to obtain grayish-white radial colonies.

[0154] (2) Seed culture expansion: Inoculate activated colonies into liquid culture medium at an inoculation rate of 6%-8%, and incubate at 40℃ and 220rpm for 36-48 hours. The viable bacterial concentration in the culture solution should be ≥10%. 9 CFU / mL terminates culture;

[0155] (3) Solid fermentation production of bacteria: inoculate the seed liquid into the fermentation substrate at an inoculation amount of 10%-12%, load into a gas permeable fermentation bag, and place in an environment of 45±2℃ and relative humidity of 70%-80% for 5-7 days. The fermentation end point characteristics are: the substrate is brownish yellow and loose, the mycelium density is ≥10 5 g under a microscope, and the viable bacterial count is ≥10 10 CFU / g. After drying, the cellulose-degrading bacterial viable bacterial preparation is obtained by crushing.

[0156] The high-temperature actinomycete strain numbered GDMCC No 4.160 Thermoactinomyces guangxiensis was purchased from the Guangdong Microbial Culture Collection Center, and the original preservation time was June 13, 2019. The strain can be purchased through the market channel.

[0157] Performance test

[0158] Antagonistic experiment: the strain of Monascus purpureus numbered CGMCC No 3.15548 and the Guangxi high-temperature actinomycete strain numbered CGMCC No 4.7156 are inoculated on CMC-Na solid culture medium as shown in Figure 1 (schematic diagram), and after 3 days of culture at 25-30℃, whether an inhibition zone appears at the intersection of different strains is observed. If there is no inhibition zone, it indicates that there is no antagonism between the strains, and the strains can be used in combination; otherwise, the strains cannot be used in combination. The culture diagram is shown in Figure 2 , and it can be seen that the two strains do not inhibit each other and can be used in combination.

[0159] Strain cellulose-degrading capacity test:

[0160] The target strains are inoculated on carboxymethyl cellulose sodium (CMC-Na) solid culture medium, 3 points are spotted per dish, and the culture dishes are inverted and cultured in a 25-30℃ constant temperature incubator. After 3 days of culture, the colonies grow out, 1.0 g / L Congo red staining solution is carefully poured into the culture dishes for staining, and the culture dishes are left to stand for 30 min. The Congo red staining solution is poured out, and then 1.0 mol / L NaCl solution is used for decolorization for 20 min. The transparent circle appearing on the plate is observed, and the diameters of the transparent circle (D) and the colony (d) are measured. The strains capable of degrading cellulose will have obvious transparent circles after treatment with Congo red staining solution, and the ratio of the transparent circle diameter D to the colony diameter d can reflect the degradation capacity. The culture results are shown in Figure 3 and Table 1.

[0161] Table 1 Ratio of transparent circle diameter D to colony diameter d of strains c

[0162]

[0163] Straw fermentation composting experiment:

[0164] CMC-Na, xylanase and laccase are biological catalysts, which directly reflect the metabolic intensity of microorganisms in the process of lignocellulose degradation. Therefore, the activities of the three enzymes in the middle of fermentation are determined to judge the fermentation effect.

[0165] The fermentation inoculant is prepared according to the method of the examples and comparative examples.

[0166] The rice straw is selected, the straw is dried and crushed into small pieces of 3-6 cm long, urea is added to adjust to about 25:1, the test inoculant is added to water and dispersed uniformly, and is uniformly sprayed on the material pile by a water sprayer, and finally the pile is turned while spraying clean water, the moisture content is adjusted to about 60%, and the mass ratio of the inoculant and the straw is 50g:10kg. The fermentation is carried out for 20 days under natural conditions, the pile is turned every 3-4 days, and after the fermentation is completed, the straw bio-organic fertilizer is obtained by drying to a moisture content of not more than 30%. Samples are taken at 3d, 10d and 20d for testing. The surface microstructure of the straw is observed at 3d, the key enzyme activity is detected at 10d, and the compost indicators are detected at 20d.

[0167] Test method:

[0168] (1) Cellulase activity determination

[0169] The cellulase enzyme activity is determined by the carboxymethyl cellulose saccharification method (Li Lianxiao, Du Jinhua, Li Junxun, et al. Research on determination of cellulase activity by CMC saccharification method. Feedstuff industry, 2006, 27(24): 49-52). The cellulase enzyme activity is defined as follows: the enzyme amount required for hydrolyzing carboxymethyl cellulose to produce 1.0 μg of glucose in 1 min under the conditions of 40 ℃ and p H 4.6 is defined as 1 enzyme activity unit (U / g).

[0170] (2) Xylanase activity determination

[0171] The xylanase enzyme activity is determined according to the method of national standard GB / T 23874-2009. The xylanase enzyme activity is defined as follows: the enzyme amount required for degrading 1 mol of reducing sugar from a xylan solution with a mass concentration of 5 mg / mL per minute under the conditions of 37 ℃ and p H 5.5 is defined as 1 enzyme activity unit (U / g).

[0172] (3) Laccase enzyme activity determination

[0173] The laccase enzyme activity was determined by ABTS oxidation method. The enzyme activity definition: the amount of enzyme required to catalyze the generation of 1 μmol ABTS・⁺ (cation radical) per minute at 30℃, pH 4.0 (reference FAN Lili, JI Jiaming, XUN Yuefeng, et al. Isolation and characterization of laccase from Trichoderma asperellum Tasjk26 [J]. Food and Fermentation Industries, 2025, 51(13): 45-52.). The enzyme activity was determined in 5 samples from different parts of the compost, and the average value was taken as the result.

[0174] Table 2 Fermentation effect experiment results

[0175]

[0176] After fermentation, the fertilizer indicators were determined, the seed germination test was conducted, and the physical and chemical indicators of the product were determined.

[0177] The sample was added with sterile distilled water at a volume ratio of 1:10, 5 mL of the compost extract was taken by a pipette gun and added to a sterile culture dish with filter paper, 10 full cabbage seeds were evenly placed on the surface, sterile water was used as a control, 3 parallel tests were conducted for each treatment, the culture dish was placed in a 25℃ constant temperature incubator for 48h, and the seed germination rate GI was determined. Generally, it is considered that the seed germination rate is more than 50% to indicate that the organic fertilizer is basically decomposed. Other related indicators are detected according to the corresponding method in the national quality standard NY525-2021 for organic fertilizer product performance. Five compost experiments were conducted for each experimental group, and the average value of the test results was taken.

[0178] Table 3 Compost test results

[0179]

[0180] From the data in Tables 1-3, it can be seen that the fermentation agent of the present application examples 1-2 shows high enzyme activity in the process of straw composting. This indicates that when the quality ratio of Aspergillus spore powder and cellulose degradation bacteria active bacteria preparation is 1:1, the combination can better promote the degradation of lignocellulose. In addition, from the data in Table 2, it can be seen that in the key enzyme activity detection of the 10th day of fermentation, the activity of cellulase and xylanase in the example 1 group also reached a high level, further verifying the superiority of the combination. FromFigure 4 The straw degradation effect diagram also shows that the surface morphology of the rice straw before degradation is smooth, well-organized, compact, and the lignocellulose structure is arranged closely without damage, and the grid structure of the parenchyma cells is complete. After the rice straw treated by the fermentation agent of Example 1 for 3 days, the wax layer on the surface of the rice straw is completely decomposed, and the surface is obviously deformed. The lignocellulose structure is obviously broken, and serious surface shrinkage occurs, and the grid integrity has been destroyed. It is very beneficial to subsequent fermentation and composting.

[0181] By analyzing the germination rate of seeds in different treatment groups, it is found that the germination rate of pakchoi seeds in Example 1 group reaches 85%, which is much higher than that of treatment groups of Comparative Examples 3 to 6, and the physicochemical indexes of the compost products meet the requirements of the national quality standard NY525-2021 for organic fertilizer. This shows that the fermentation agent can not only effectively improve the composting efficiency, but also ensure the safety and fertilizer efficiency of the compost product. The comparative examples 1-6 change the composition of the strains, the synergistic balance of the key strains is broken, and the enzyme activity decreases, which leads to the weakening of the fermentation effect. Based on the above experimental results, the biological organic fertilizer fermentation agent provided by the present application has significant technical advantages, especially in improving enzyme activity, accelerating straw degradation and improving the quality of compost products, and has good application prospect.

[0182] It should be noted that the above examples are only part of the preferred modes of implementing the present application, not all. Obviously, based on the above examples of the present application, all other examples obtained by those of ordinary skill in the art without creative labor should belong to the scope of protection of the present application.

Claims

1. A bio-organic fertilizer fermentation agent, characterized in that, The microbial agent comprises active microorganisms and a synergistic component, and the mass ratio of the two is 1:(0.3-0.6); the active microorganisms comprise aspergillus spore powder and cellulose-degrading bacteria active bacteria preparation, and the mass ratio of the two is 1:1; the preparation method of the aspergillus spore powder is as follows: (1) inoculate the Monascus purpureus with the strain number of CGMCC No 3.15548 into an activated culture medium A, and incubate at a constant temperature of 25±2 DEG C for 3-5 days to obtain activated slant bacteria; (2) the activated bacteria are inoculated into a liquid seed culture medium, and cultured at a shaking speed of 220-250 rpm and at 23-25°C for 48-72 hours until the concentration of the bacteria in the bacterial liquid is ≥10 8 CFU / mL, to obtain a seed liquid; (3) the seed liquid is inoculated into solid sporulation medium at 5%-8% inoculation amount, and then is cultured at 23-25°C and relative humidity of 75%-85% in the dark for 7-10 days until the surface of the medium is covered with a red spore layer and the spore density is ≥10 9 spores / g under a microscope; (4) add sterile water containing 0.05% Tween-80 into the spore-producing substrate, stir and elute the spores, remove the mycelium residues through 300-mesh filter cloth, centrifuge the filtrate at 3000 rpm for 10 minutes at 4 DEG C to collect the spore precipitate, and adopt vacuum freeze-drying treatment to obtain aspergillus spore powder with a water content of less than or equal to 8% and a spore survival rate of greater than or equal to 90%; The preparation method of the cellulose-degrading bacteria active bacteria preparation is as follows: (1) strain screening and activation: inoculate the Guangxi high-temperature actinomycete with the strain number of CGMCC No 4.7156 into an activated culture medium B, and incubate in a constant-temperature incubator at 42±2 DEG C in the dark for 48-72 hours to obtain gray-white radial colonies; (2) Seed liquid expansion culture: inoculate the activated colonies into liquid culture medium with an inoculation amount of 6%-8%, and place in a 40°C, 220 rpm shaker for culture for 36-48 hours, and the viable bacterial concentration of the bacterial liquid is ≥10 9 CFU / mL to terminate the culture; (3) solid fermentation production bacteria: inoculate the seed liquid into the fermentation substrate at 10%-12% inoculation amount, load into a gas permeable fermentation bag, and place in an environment of 45±2°C and relative humidity of 70%-80% for 5-7 days. The fermentation end point characteristics are: the substrate is brownish yellow and loose, the mycelium density is ≥10 5 / cm under a microscope, and the viable bacterial count is ≥10 10 CFU / g. After drying, the cellulose-degrading bacteria viable bacterial preparation is obtained.

2. The bio-organic fertilizer fermentation agent according to claim 1, characterized in that, The synergistic component comprises the following raw materials in parts by mass: wood ash 30-40 parts, bone meal 5-10 parts, and brown sugar 10-15 parts.

3. The preparation method of the bio-organic fertilizer fermentation agent according to any one of claims 1-2, characterized in that, The method comprises the following steps: (1) prepare the aspergillus spore powder and the cellulose-degrading bacteria active bacteria preparation, and mix the two according to a mass ratio of 1:1; (2) mix the active microorganisms and the synergistic component according to the proportion, first sieve the wood ash and the bone meal in the synergistic component to 80-100 mesh to ensure uniform particle size; then add brown sugar and an appropriate amount of water into a stirring device to dissolve and form a sugar solution; uniformly spray the sugar solution on the surface of the wood ash and bone meal mixture, add the active microorganisms while stirring, control the stirring speed to be 60-80 rpm, and control the time to be 5-10 minutes, so that the components are fully mixed; after the mixing is completed, place the material in an environment with a temperature of 25-30 DEG C and a relative humidity of 50%-60% for 1-2 hours to promote the adaptation of the microorganisms to the environment and preliminary activation; finally, pack the mixed microbial agent into a sealed packaging bag, and adopt vacuum packaging technology to avoid external pollution.

4. The use of the bio-organic fertilizer fermentation agent according to any one of claims 1-2, characterized in that, The method is used for straw fermentation to prepare organic fertilizer.

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