CN117866817A - Straw decomposition agent and method for decomposing corn straw - Google Patents
Straw decomposition agent and method for decomposing corn straw Download PDFInfo
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- CN117866817A CN117866817A CN202311808558.8A CN202311808558A CN117866817A CN 117866817 A CN117866817 A CN 117866817A CN 202311808558 A CN202311808558 A CN 202311808558A CN 117866817 A CN117866817 A CN 117866817A
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Classifications
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02W—CLIMATE CHANGE MITIGATION TECHNOLOGIES RELATED TO WASTEWATER TREATMENT OR WASTE MANAGEMENT
- Y02W30/00—Technologies for solid waste management
- Y02W30/40—Bio-organic fraction processing; Production of fertilisers from the organic fraction of waste or refuse
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- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The invention provides a straw decomposition agent and a method for decomposing corn straw. The straw decomposition agent comprises 18-30 parts of aspergillus awamori preparation, 25-40 parts of bacillus subtilis preparation and 15-25 parts of trichoderma reesei preparation. The straw decomposition agent can accelerate the decomposition speed of corn straw under the low-temperature condition, improve the soil fertility, increase the soil water content, reduce the soil capacity, improve the soil porosity, loosen the soil and increase the air permeability. Effectively solves the problems of slow decomposition, low decomposition rate and incomplete decomposition of the corn straw at low temperature.
Description
Technical Field
The invention relates to the technical field of crop straw degradation, in particular to a straw decomposition agent and a method for decomposing corn straw.
Background
Straw returning is an important way for balancing soil carbon loss and supplementing soil organic matters, and is the most main mode of resource utilization. After returning the straw to the field, the straw can be decomposed and converted into humus under the action of soil microorganisms; the main supporting tissue of the straw is lignocellulose, is a main object of microbial degradation, and after the straw is returned to the field, nutrients in the straw are released by decay explanation, so that the soil fertility can be improved, the agricultural yield can be directly promoted, the fertilizer utilization rate can be improved, the contradiction between fertilizer supply and demand can be relieved, the input-output ratio can be reduced, the agricultural production cost can be reduced, and the sustainable development of agriculture can be facilitated.
However, the main components of the straw are cellulose and hemicellulose, and the lignin content is inferior, and the three main components form a lignocellulose structure which is difficult to be decomposed by microorganisms in a natural state. Cellulose, hemicellulose and lignin in the straw have compact structure and strong decomposition resistance, so that the straw is slowly degraded under natural conditions and is difficult to cultivate. Especially in northern areas, the temperature is rapidly reduced after autumn, the rainfall is reduced, and especially after 11 months, the corn stalk decomposition speed is quite slow. The corn stalk decomposition rate is low during spring sowing, and the problems of large soil surface layer gap, low emergence rate, dead seedling, weak seedling and the like can be generated.
Disclosure of Invention
In order to solve the problems of slow decomposition speed and incomplete decomposition of corn stalks under the low-temperature condition, the invention provides a stalk decomposition agent which can accelerate the decomposition speed of corn stalks and is beneficial to improving the physical and chemical properties of soil and a method for decomposing corn stalks.
In order to achieve the above object, the present invention provides the following technical solutions:
the invention provides a straw decomposition agent which comprises the following components in parts by weight: 18 to 30 parts of aspergillus awamori preparation, 25 to 40 parts of bacillus subtilis preparation and 15 to 25 parts of trichoderma reesei preparation;
the effective viable count in the aspergillus awamori preparation, the bacillus subtilis preparation and the trichoderma reesei preparation is independently 1-5 multiplied by 10 9 cfu/g。
Preferably, the straw decomposition agent further comprises auxiliary materials, wherein the auxiliary materials comprise one or more of calcium carbonate, turfy soil, bean pulp, wheat bran, rice hull powder and straw powder; the weight portion of the auxiliary material is 10-20 portions. The straw decomposition agent is added with auxiliary materials to protect thalli and reduce the influence of factors which are unfavorable for the survival of the thalli in the environment.
The invention also provides a method for decomposing the corn straw, which comprises the following steps:
(1) Activating the straw degrading bacterial agent to obtain bacterial liquid;
(2) Crushing corn straw, inoculating the bacterial liquid in the step (1), applying urea, and turning the straw and the urea into soil for decomposition.
Preferably, the spore concentration in the bacterial liquid is 1-9×10 7 cfu/mL。
Preferably, the step of activating the straw decomposition agent comprises the following steps: inoculating straw decomposition agent into an activation culture medium, standing and culturing for 5-8 days at 20-30 ℃, filtering the culture to obtain spore suspension, and regulating the spore concentration in the spore suspension to 1-9 multiplied by 10 by using sterile water 7 cfu/mL to obtain the bacterial liquid.
Preferably, the activation medium is: 10-15 g of cellobiose, 5-10 g of peptone, 3-5 g of yeast powder, 0.3-1 g of urea and water to 1000mL; ph=6.5 to 7.0.
Preferably, the inoculation mode of the bacterial liquid is spraying inoculation, and the spraying amount of the bacterial liquid is 300-800 kg/hm 2 。
Preferably, the corn stalk is crushed to a grain size of 1-2 cm, and the returning amount of the corn stalk is 2000-3500 kg/hm 2 。
Preferably, the depth of the corn stalk buried in the soil is 10-15 cm.
Preferably, the urea is applied in an amount of 200 to 400kg/hm 2 The method comprises the steps of carrying out a first treatment on the surface of the The water quantity is 150 to 250t/hm 2 。
By adopting the technical scheme, the invention has the following beneficial effects: the straw decomposition agent can accelerate the decomposition speed of corn straw under the low-temperature condition, improve the soil fertility, increase the soil water content, reduce the soil capacity, improve the soil porosity, loosen the soil and increase the air permeability. The depth of the corn stalk buried in the soil is 10-15 cm, and in the depth range, the corn stalk has the best decomposition effect, so that the problems of slow decomposition, low decomposition rate and incomplete decomposition of the corn stalk at low temperature are effectively solved.
Detailed Description
The invention provides a straw decomposition agent which comprises the following components in parts by weight: 18 to 30 parts of aspergillus awamori preparation, 25 to 40 parts of bacillus subtilis preparation and 15 to 25 parts of trichoderma reesei preparation;
the aspergillus awamori preparation, the bacillus subtilis preparation and the trichoderma reesei preparationThe number of effective viable bacteria in the culture medium is independently 1-5 multiplied by 10 9 cfu/g。
In the straw decomposition agent, the weight part of the aspergillus awamori preparation is 18-30 parts, more preferably 23-27 parts, and still more preferably 25 parts;
the weight part of the bacillus subtilis preparation is 25-40 parts, more preferably 28-36 parts, and even more preferably 33 parts;
the weight part of the trichoderma reesei preparation is 15-25 parts, more preferably 17-22 parts, and still more preferably 20 parts.
In the invention, the aspergillus awamori can generate feruloyl esterase, ferulic acid is often crosslinked with cell wall polysaccharide through an ether bond and lignin to form a complex network structure, the ferulic acid is dissociated from the structure of the plant cell wall to destroy the skeleton structure of the cell wall, the connection among lignin, hemicellulose and cellulose is partially broken, the structure is loosened, and the decomposition of straw is promoted;
the bacillus subtilis can secrete rich cellulase and promote decomposition of organic matters such as cellulose and hemicellulose;
the trichoderma reesei is a filamentous fungus, secretes cellulase and can efficiently decompose cellulose and lignin;
the invention cultures aspergillus awamori, bacillus subtilis and trichoderma reesei together, the overall growth condition of the composite microbial inoculum is obviously improved, the straw degradation capability is further improved, the extracellular metabolites of the three bacteria have activation promotion effects on respective corresponding degradation enzymes, and the compatibility of the three bacterial strains can synergistically improve the straw degradation efficiency. Meanwhile, aspergillus awamori, bacillus subtilis and trichoderma reesei have an inhibiting effect on harmful bacteria in the process of degrading straw, and the degradation rate of the corn straw is increased, so that the carried bacteria of the corn straw are correspondingly reduced, and after the degraded corn straw is returned to the field, the field diseases can be effectively inhibited. The aspergillus awamori, the bacillus subtilis and the trichoderma reesei of the invention co-act, and can efficiently and synergistically degrade the corn stalks at a lower temperature.
The straw decomposition agent also comprises auxiliary materials, wherein the auxiliary materials comprise one or more of calcium carbonate, turfy soil, bean pulp, wheat bran, rice hull powder and straw powder; the weight portion of the auxiliary material is 10-20 portions, more preferably 13-17 portions, and even more preferably 15 portions.
In the invention, the preparation methods of the aspergillus awamori preparation, the bacillus subtilis preparation and the trichoderma reesei preparation are as follows:
(1) Activation of
Inoculating Aspergillus awamori, bacillus subtilis and Trichoderma reesei in the activating culture medium, and standing for culturing to obtain fully activated Aspergillus awamori, bacillus subtilis and Trichoderma reesei.
The formula of the activation culture medium is as follows: 10-15 g of cellobiose, 5-10 g of peptone, 3-5 g of yeast powder, 0.3-1 g of urea and water to 1000mL; ph=6.5 to 7.0.
The temperature of the stationary culture is 20 to 30 ℃, more preferably 22 to 27 ℃, and still more preferably 25 ℃; the stationary culture time is 25 to 35 hours, more preferably 27 to 32 hours, and still more preferably 30 hours.
The formulation of the activation medium is further preferably: 12-14 g of cellobiose, 7-9 g of peptone, 3.4-4.5 g of yeast powder, 0.5-0.8 g of urea and water to 1000mL; ph=6.7 to 6.9; further preferred are: 13g of cellobiose, 8g of peptone, 4g of yeast powder, 0.6g of urea and water to 1000mL; ph=6.5.
(2) Preparation of microbial preparations
Inoculating the strains fully activated in the step (1) into a seed culture medium respectively, carrying out shaking culture (seed fermentation), and inoculating into a fermentation tank of an expansion culture medium, wherein the components of the expansion culture medium are the same as those of the seed culture medium; fermenting the same seed under the fermentation condition, centrifuging the fermentation liquid, collecting the centrifuged precipitate thallus, adding a freeze-drying protective agent, and performing freeze-drying treatment to prepare each microbial preparation.
The inoculum size of the bacterial liquid in the step (2) is 7-15%, more preferably 9-13%, and even more preferably 11%; the temperature of the shaking culture is 20 to 30 ℃, more preferably 22 to 27 ℃, still more preferably 25 ℃; the time of the shaking culture is 45 to 50 hours, more preferably 46 to 49 hours, still more preferably 48 hours; the rotation speed of the shaking culture is 100 to 200rpm, more preferably 120 to 180rpm, still more preferably 150rpm; after shaking culture, carrying out expansion culture, wherein the inoculum size, culture temperature, rotation speed and culture medium components of the expansion culture are the same as those of seed fermentation, and the time of the expansion culture is 60-72 h, more preferably 64-70 h, still more preferably 67h; after completion of the expansion culture, the fermentation broth is centrifuged at 7000 to 9000rpm, more preferably 7500 to 8500rpm, still more preferably 8000rpm; the centrifugation time is 8 to 13 minutes, more preferably 9 to 11 minutes, and still more preferably 10 minutes. Collecting the centrifuged precipitate thallus, adding freeze-drying protective agent, and freeze-drying to obtain each microbial preparation. The freeze-drying protective agent is preferably one or more of glycerol, mannitol, sorbitol and trehalose; the addition amount of the freeze-drying protective agent is 0.2-0.5%, more preferably 0.3-0.4%, and still more preferably 0.35% of the weight of the thallus; the temperature of the freeze-drying is-85 to-75 ℃, more preferably-83 to-78 ℃, and still more preferably-80 ℃. After Aspergillus awamori, bacillus subtilis and Trichoderma reesei are obtained, the straw decomposition agent is obtained by combining the materials in parts by weight.
The formula of the seed culture medium is as follows: 10 to 20g of corn straw powder, 3 to 8g of peptone, 0.5 to 1.5g of yeast extract, 3 to 7g of NaCl and K 2 HPO 4 0.5～1.5g、MgSO 4 ·7H 2 O 0.3～0.5g、CaCO 3 2 to 5g of water is added to 1000mL, and pH=6.5 to 7.0, more preferably: 12-18 g of corn straw powder, 4-6 g of peptone, 0.8-1.2 g of yeast extract, 4-6 g of NaCl and K 2 HPO 4 0.7～1.3g、MgSO 4 ·7H 2 O 0.35～0.45g、CaCO 3 3-4 g, water is added to 1000mL, and pH=6.6-6.8; more preferably: 15g of corn stalk powder, 5g of peptone, 1g of yeast extract, 5g of NaCl and K 2 HPO 4 1g、MgSO 4 ·7H 2 O 0.4g、CaCO 3 3.5g, water was made up to 1000ml, ph=6.5.
The corn stalk powder is obtained by drying corn stalk until the water content is below 1%, crushing the corn stalk powder and sieving the crushed corn stalk powder with a 30-mesh sieve, wherein the drying temperature is 60-70 ℃, more preferably 62-67 ℃, and still more preferably 65 ℃.
The invention also provides a method for decomposing the corn straw, which comprises the following steps:
(1) Activating the straw degrading bacterial agent to obtain bacterial liquid;
(2) Crushing corn straw, inoculating the bacterial liquid in the step (1), applying urea, and turning the straw and the urea into soil for decomposition.
The straw degrading bacterial agent is activated to obtain bacterial liquid; the microbial inoculum of the invention comprises the following activation steps: inoculating the microbial inoculum into an activation culture medium, standing and culturing for 5-8 days at 20-30 ℃, filtering the culture to obtain spore suspension, and regulating the spore concentration in the spore suspension to 1-9 multiplied by 10 by using sterile water 7 cfu/mL to obtain the bacterial liquid.
The ratio of the inoculation amount of the degradation microbial inoculum to the weight volume of the activation culture medium is 3-5 g/100 mL, more preferably 3.5-4.5 g/100 mL, and still more preferably 4 g/100 mL; after inoculation, carrying out stationary culture, wherein the temperature of the stationary culture is 20-30 ℃, more preferably 22-27 ℃, and still more preferably 25 ℃; the stationary culture time is 5 to 8 days, more preferably 6 to 7 days, still more preferably 6.5 days; filtering the culture to obtain spore suspension, and regulating spore concentration in the spore suspension to 1-9×10 with sterile water 7 cfu/mL to obtain the bacterial liquid. The spore concentration in the bacterial liquid is 1-9 multiplied by 10 7 cfu/mL, more preferably 3 to 7X 10 7 cfu/mL, more preferably 5X 10 7 cfu/mL。
The formula of the activation culture medium used when decomposing the corn stalks is the same as that of the activation culture medium used in the preparation stage of the microbial inoculum.
The invention pulverizes the straw, inoculates the bacterial liquid after activation, and apply urea, turn over straw, urea into soil and carry on the decomposition. The grain size of the crushed straw is 1-2 cm, more preferably 1.2-1.7 cm, and even more preferably 1.5cm; the returning amount of the straw is 2000-3500 kg/hm 2 Further preferably 2300 to 3200kg/hm 2 More preferably 2600kg/hm 2 The method comprises the steps of carrying out a first treatment on the surface of the The inoculation mode of the bacterial liquid is spraying inoculation, and the spraying amount of the bacterial liquid is 300-800 kg/hm 2 Further preferably 400 to 600kg/hm 2 More preferably 500kg/hm 2 The method comprises the steps of carrying out a first treatment on the surface of the The application amount of the urea is 200-400 kg/hm 2 More preferably 250 to 350kg/hm 2 More preferably 300kg/hm 2 The urea is added when the corn stalks are decomposed, so that the decomposing and decomposing of the microorganisms on the corn stalks are accelerated. The microorganism takes carbon as an energy source, nitrogen is taken as nutrition, the carbon-nitrogen ratio suitable for microorganism activity is 25:1, the carbon-nitrogen ratio of the straw is 65-85:1, and the nitrogen in the soil is deficient after the straw is returned to the field, so that the microorganism competes for the nitrogen with crops, the carbon substances in the soil can be increased after the corn stalk scale is directly returned to the field, and the nitrogen is absorbed from the soil when the microorganism decomposes the carbon. If the nitrogen fertilizer is not applied in time, insufficient nitrogen in soil is easily caused, the growth of the next crop is affected, and even the yield is reduced. Therefore, the invention can not only rapidly decompose the straw, but also ensure the normal growth of crops in the straw returning Shi Zengshi nitrogenous fertilizer. Watering after turning over, wherein the watering quantity is 150-250 t/hm 2 Further preferably 180 to 220t/hm 2 More preferably 200t/hm 2 。
According to the invention, straw and urea are buried into soil for decomposition, the depth of the corn straw buried into the soil is 10-15 cm, more preferably 12-14 cm, and even more preferably 13cm, and under the depth, the rapid decomposition of the straw is facilitated, and the returning effect is good.
The technical solutions provided by the present invention are described in detail below with reference to examples, but they should not be construed as limiting the scope of the present invention.
In the embodiment of the invention, the Paoshengqu enzyme is purchased from China general microbiological culture Collection center (CGMCC) with the preservation number of CGMCC No.18595;
bacillus subtilis is purchased from China center for type culture collection of agricultural microorganisms, and the preservation number is ACCC 19374;
trichoderma reesei was purchased from China center for type culture Collection with a collection number of CCTCC DF 20081039.
Example 1
1. Preparation of microbial inoculum
(1) Activating strains: respectively inoculating Aspergillus awamori, bacillus subtilis and Trichoderma reesei in the activating culture medium, and standing at 20deg.C for 25 hr to obtain activated strain.
The formula of the activation culture medium is as follows: 10g of cellobiose, 5g of peptone, 3g of yeast powder, 0.3g of urea and water to 1000mL; ph=6.5.
(2) Seed fermentation: inoculating the fully activated strains into a seed culture medium according to the inoculation amount of 75%, and carrying out shaking culture at 200 ℃ and 100rpm for 45 hours to obtain seed fermentation liquor;
the formula of the seed culture medium is as follows: 10g of corn stalk powder, 3g of peptone, 0.5g of yeast extract, 3g of NaCl and K 2 HPO 4 0.5g、MgSO 4 ·7H 2 O 0.3g、CaCO 3 25g, water to 1000ml, ph=6.5; the corn stalk powder is obtained by drying corn stalk at 60deg.C until the water content is below 1%, pulverizing, and sieving with 30 mesh sieve;
(3) And (3) performing expansion culture: inoculating the seed fermentation liquor into a new culture medium according to the inoculation amount of 7% for expansion culture, wherein the time of the expansion culture is 60 hours, and the temperature, the rotating speed and the culture medium components of the expansion culture are the same as those of the seed fermentation;
(4) And (3) preparation of a microbial inoculum: after the expansion culture is completed, the fermentation liquor is centrifuged at 7000rpm for 8min, then the thalli are collected, glycerin with the weight of 0.2 percent of the thalli is added and freeze-dried at the temperature of minus 85 ℃ to respectively obtain an aspergillus awamori preparation, a bacillus subtilis preparation and a trichoderma reesei preparation, and the effective viable count in the aspergillus awamori preparation, the bacillus subtilis preparation and the trichoderma reesei preparation is 1.8x10 respectively 9 cfu/g、2.3×10 9 cfu/g、2.1×10 9 cfu/g。
Mixing 18kg of Aspergillus awamori preparation, 25kg of Bacillus subtilis preparation, 15kg of Trichoderma reesei preparation and 10kg of turfy soil to obtain the straw decomposition agent.
2. Decomposition corn stalk
3kg of the prepared straw decomposition agentInoculating into 100L activating culture medium, standing at 20deg.C for 5 days, filtering to obtain spore suspension, and regulating spore concentration in spore suspension to 1×10 with sterile water 7 cfu/mL to obtain the bacterial liquid. The formula of the activation culture medium used when decomposing the corn straw is the same as that of the activation culture medium used in the preparation stage of the microbial inoculum.
Crushing the straw to the grain size of 1cm, spraying and inoculating the activated bacterial liquid, applying urea, turning the straw and the urea into soil, and watering to carry out decomposition. The returning amount of the straw is 2000kg/hm 2 The spraying amount of the bacterial liquid is 300kg/hm 2 The urea was applied in an amount of 200kg/hm 2 The depth of the corn stalk buried in the soil is 10cm, and the watering amount is 150t/hm 2 。
Example 2
1. Preparation of microbial inoculum
(1) Activating strains: respectively inoculating Aspergillus awamori, bacillus subtilis and Trichoderma reesei in the activating culture medium, and standing at 25deg.C for 30 hr to obtain activated strain.
The formula of the activation culture medium is as follows: 12g of cellobiose, 8g of peptone, 4g of yeast powder, 0.6g of urea and water to 1000mL; ph=6.8.
(2) Seed fermentation: inoculating the fully activated strains into a seed culture medium according to the inoculum size of 9%, and carrying out shaking culture for 48 hours at 25 ℃ and 150rpm to obtain seed fermentation liquor;
the formula of the seed culture medium is as follows: 15g of corn stalk powder, 6g of peptone, 1g of yeast extract, 5g of NaCl and K 2 HPO 4 1g、MgSO 4 ·7H 2 O 0.4g、CaCO 3 3g, water to 1000ml, ph=6.8; the corn stalk powder is obtained by drying corn stalk at 65deg.C until the water content is below 1%, pulverizing, and sieving with 30 mesh sieve;
(3) And (3) performing expansion culture: inoculating the seed fermentation liquor into a new culture medium according to the inoculation amount of 9% for expansion culture, wherein the time of the expansion culture is 65h, and the temperature, the rotating speed and the culture medium components of the expansion culture are the same as those of the seed fermentation;
(4) And (3) preparation of a microbial inoculum: completion of the expansion cultureAfter centrifugation of the fermentation liquor at 8000rpm for 10min, collecting thalli, adding mannitol with the weight of 0.3% of the weight of the thalli, and freeze-drying at-80 ℃ to respectively obtain an aspergillus awamori preparation, a bacillus subtilis preparation and a trichoderma reesei preparation, wherein the effective viable count in the aspergillus awamori preparation, the bacillus subtilis preparation and the trichoderma reesei preparation is 2.6x10 respectively 9 cfu/g、3.1×10 9 cfu/g、2.8×10 9 cfu/g。
25kg of Aspergillus awamori preparation, 33kg of Bacillus subtilis preparation, 18kg of Trichoderma reesei preparation and 15kg of soybean meal are mixed to obtain the straw decomposition agent.
2. Decomposition corn stalk
Inoculating 4kg of the prepared straw decomposition agent into 100L of activating culture medium, standing at 25deg.C for 7 days, filtering to obtain spore suspension, and regulating spore concentration in spore suspension to 5×10 with sterile water 7 cfu/mL to obtain the bacterial liquid. The formula of the activation culture medium used when decomposing the corn straw is the same as that of the activation culture medium used in the preparation stage of the microbial inoculum.
Crushing the straw to the grain size of 1.5cm, spraying and inoculating the activated bacterial liquid, applying urea, turning the straw and the urea into soil, and watering to carry out decomposition. The returning amount of the straw is 2800kg/hm 2 The spraying amount of the bacterial liquid is 500kg/hm 2 Urea was applied in an amount of 300kg/hm 2 The depth of the corn straw buried in the soil is 13cm, and the watering amount is 200t/hm 2 。
Example 3
1. Preparation of microbial inoculum
(1) Activating strains: respectively inoculating Aspergillus awamori, bacillus subtilis and Trichoderma reesei in the activating culture medium, and standing at 30deg.C for 35 hr to obtain activated strain.
The formula of the activation culture medium is as follows: 15g of cellobiose, 10g of peptone, 5g of yeast powder, 1g of urea and water to 1000mL; ph=7.0.
(2) Seed fermentation: inoculating the fully activated strains into a seed culture medium according to 15% of inoculum size, and carrying out shaking culture at 30 ℃ and 200rpm for 50 hours to obtain seed fermentation liquor;
the formula of the seed culture medium is as follows: 20g of corn straw powder, 8g of peptone, 1.5g of yeast extract, 7g of NaCl and K 2 HPO 4 1.5g、MgSO 4 ·7H 2 O 0.5g、CaCO 3 5g, water to 1000ml, ph=7.0; the corn stalk powder is obtained by drying corn stalk at 70deg.C until the water content is below 1%, pulverizing, and sieving with 30 mesh sieve;
(3) And (3) performing expansion culture: inoculating the seed fermentation liquor into a new culture medium according to 15% of inoculum size for expansion culture, wherein the time of the expansion culture is 72 hours, and the temperature, the rotating speed and the culture medium components of the expansion culture are the same as those of the seed fermentation;
(4) And (3) preparation of a microbial inoculum: after the expansion culture is completed, the fermentation liquor is centrifuged at 9000rpm for 8-13 min, then the thalli are collected, glycerol, mannitol, sorbitol and trehalose which are 0.5 percent of the weight of the thalli are added, and freeze-dried at the temperature of minus 75 ℃ to respectively obtain an aspergillus awamori preparation, a bacillus subtilis preparation and a trichoderma reesei preparation, and the effective viable bacteria number in the aspergillus awamori preparation, the bacillus subtilis preparation and the trichoderma reesei preparation is 3.5x10 respectively 9 cfu/g、4.2×10 9 cfu/g、3.7×10 9 cfu/g。
Mixing 30kg of aspergillus awamori preparation, 40kg of bacillus subtilis preparation, 25kg of trichoderma reesei preparation and 20kg of wheat bran soil to obtain the straw decomposition agent.
2. Decomposition corn stalk
Inoculating 5kg of the stalk decomposition agent into 100L of activating culture medium, standing at 30deg.C for 8 days, filtering to obtain spore suspension, and regulating spore concentration in spore suspension to 9×10 with sterile water 7 cfu/mL to obtain the bacterial liquid. The formula of the activation culture medium used when decomposing the corn straw is the same as that of the activation culture medium used in the preparation stage of the microbial inoculum.
Crushing the straw to the grain size of 2cm, spraying and inoculating the activated bacterial liquid, applying urea, turning the straw and the urea into soil, and watering to carry out decomposition. Returning straw to field of 3500kg/hm 2 The spraying amount of the bacterial liquid is 800kg/hm 2 The urea was applied in an amount of 4000kg/hm 2 Corn stalk is buried in soilIs 15cm deep, and the water quantity is 250t/hm 2 。
Comparative example 1
Unlike example 1, the species of straw decomposition agent of comparative example 1 contained only an aspergillus awamori formulation.
Comparative example 2
Unlike example 1, the species of straw decomposition agent of comparative example 1 contained only bacillus subtilis preparation.
Comparative example 3
Unlike example 1, the species of straw decomposition agent of comparative example 1 contained only trichoderma reesei preparation.
Comparative example 4
Unlike example 1, the species of straw decomposition agent of comparative example 1 contained only an aspergillus awamori formulation and a bacillus subtilis formulation.
Comparative example 5
Unlike example 1, the species of straw decomposition agent of comparative example 1 contained only an aspergillus awamori formulation and a trichoderma reesei formulation.
Comparative example 6
Unlike example 1, the species of straw decomposition agent of comparative example 1 contained only bacillus subtilis and trichoderma reesei preparations.
Comparative example 7
Unlike example 1, the straw was directly spread on the surface of the soil and covered for returning to the field.
Comparative example 8
Unlike example 1, the straw was uniformly mixed into 15-30 cm soil.
Experimental example 1
The experiment was performed in the land of principals at 11 months 2022 to 3 months 2023. The average air temperature in the local area during the experiment was around-8 ℃.
Frame planting experiment: the plastic barrel has a diameter of 30cm and a barrel depth of 25cm, and the bottom of the barrel is sawed off. And (5) manually digging a soil pit with the same size as the frame planting barrel in the field, and putting the plastic barrel into the soil pit. The dry mass of the straw scale is 100 g/basin, and the straw scale is baked to constant weight at 55 ℃. 10 pots were treated for a total of 110 pots. Examples 1 to 3 and comparative examples 1 to 6 were mixed with 15g of the activated bacteria liquid, 10g of urea and a straw scale, and then placed in a nylon mesh bag and embedded in a frame cultivation having a depth of 10 to 15cm. Comparative example 7 straw was laid flat on the ground, comparative example 8 nylon bags were buried in a 20-30 cm deep frame, and then 1kg of water was poured over the diameter of the barrels, respectively.
Micro-area experiment: performing field micro-area test at field experiment station, wherein each cell is 1m 2 After removing the soil layer with the surface of 10-15 cm, uniformly laying 200g (l m in the field) 2 Actual straw returning amount) and 20g of urea, then spraying 30g of the activated straw scale decomposed bacterial liquid, and then pressing the excavated soil to be equivalent to mechanical turning-over, 11 treatments, repeating for 3 times and arranging randomly. In the comparative example 7, the straws are paved on the land surface, the straw in the comparative example 8 is buried at a distance of 20-30 cm from the land surface, the control group only adds urea with the same mass, and then 15kg of water is poured into each cell.
Frame planting experiments on days 10, 20 and 30 after the experiments are carried out, randomly taking 2 frame planting nylon net bags in each group, measuring dry mass after drying the straws, calculating straw decomposition rate and taking an average value. Straw scale decomposition rate% = straw post decomposition dry mass/pre decomposition dry mass, straw decomposition rates for different groups are shown in table 1.
Table 1 decomposition rates of straw
| Group of | 10d(％) | 20d(％) | 30d(％) |
| Example 1 | 28.3 | 57.6 | 70.2 |
| Example 2 | 30.5 | 59.2 | 72.3 |
| Example 3 | 31.2 | 60.9 | 75.6 |
| Comparative example 1 | 8.3 | 19.2 | 25.4 |
| Comparative example 2 | 6.4 | 15.1 | 20.6 |
| Comparative example 3 | 10.7 | 22.6 | 27.8 |
| Comparative example 4 | 17.5 | 40.5 | 54.7 |
| Comparative example 5 | 20.3 | 47.2 | 58.5 |
| Comparative example 6 | 18.6 | 43.6 | 52.6 |
| Comparative example 7 | 15.5 | 35.2 | 45.9 |
| Comparative example 8 | 22.3 | 50.7 | 58.1 |
As is clear from Table 1, at 10 days of the frame cultivation experiment, the decomposition rates of the Aspergillus awamori preparation, the Bacillus subtilis preparation and the Trichoderma reesei preparation on the corn straw are 8.3%, 6.4% and 10.7%, respectively, while the decomposition rates of the corn straw in examples 1 to 3 are 28.3%, 30.5% and 31.2% respectively, which are higher than the decomposition rates of the Aspergillus awamori preparation, the Bacillus subtilis preparation and the Trichoderma reesei preparation on the corn straw, respectively, so that the Aspergillus awamori preparation, the Bacillus subtilis preparation and the Trichoderma reesei preparation have synergistic effects, accelerate the decomposition of the corn straw, and have the best decomposition effect when the straw is buried in the ground by 10-15 cm.
In the micro-area test, 10-15 cm soil samples of each treatment area are randomly collected in the field according to an X method in 2023 month, the water content of the soil (a drying method) and the volume weight of the soil (a cutting ring method) are measured, 1kg of soil samples are reserved for analysis by a four-division method after a part of samples are air-dried and fully mixed, and the results of measuring quick-acting phosphorus, alkaline nitrogen, organic matters and quick-acting potassium of the soil are shown in tables 2 and 3 respectively.
TABLE 2 soil fertility for different groups
As shown in Table 2, the straw decomposing inoculant can effectively improve the contents of organic matters, alkaline hydrolysis nitrogen, quick-acting phosphorus and quick-acting potassium in soil, and respectively improve the contents of the quick-acting phosphorus, the alkaline hydrolysis nitrogen, the organic matters and the quick-acting potassium in the soil by more than 35.75%, 21.51%, 24.71 and 24.59%.
TABLE 3 soil moisture content and soil Capacity for different groups
As can be seen from table 3, the straw decomposition agent of the present invention can increase the water content of soil, reduce the soil capacity, increase the soil porosity, loosen the soil and increase the air permeability. The straw scale is returned to the field to reduce the evaporation rate of soil moisture, reduce the evaporation capacity, strengthen the water storage and soil moisture conservation capacity of the soil, and create proper soil moisture conditions for the growth of the next crop.
From the above examples, the present invention provides a straw decomposition agent and a method for decomposing corn straw. The straw decomposition agent can accelerate the decomposition speed of corn straw under the low-temperature condition, improve the soil fertility, increase the soil water content, reduce the soil capacity, improve the soil porosity, loosen the soil and increase the air permeability.
The foregoing is merely a preferred embodiment of the present invention and it should be noted that modifications and adaptations to those skilled in the art may be made without departing from the principles of the present invention, which are intended to be comprehended within the scope of the present invention.
Claims (10)
1. The straw decomposition agent is characterized by comprising the following components in parts by weight: 18 to 30 parts of aspergillus awamori preparation, 25 to 40 parts of bacillus subtilis preparation and 15 to 25 parts of trichoderma reesei preparation;
the aspergillus awamori preparation and the bacillus subtilisThe effective viable count in the bacillus preparation and the Trichoderma reesei preparation is independently 1 to 5 multiplied by 10 9 cfu/g。
2. The straw decomposition agent according to claim 1, further comprising an auxiliary material, wherein the auxiliary material comprises one or more of calcium carbonate, peatmoss, bean pulp, wheat bran, rice hull powder, straw powder; the weight portion of the auxiliary material is 10-20 portions.
3. The method for decomposing the corn stalks is characterized by comprising the following steps of:
(1) Activating the straw decomposition agent according to claim 1 or 2 to obtain bacterial liquid;
(2) Crushing corn straw, inoculating the bacterial liquid in the step (1), applying urea, turning the straw and the urea into soil, and watering to decompose.
4. The method according to claim 3, wherein the spore concentration in the bacterial liquid is 1 to 9X 10 7 cfu/mL。
5. A method according to claim 3, wherein the step of activating the straw-degrading agent is: inoculating straw decomposition agent into an activation culture medium, standing and culturing for 5-8 days at 20-30 ℃, filtering the culture to obtain spore suspension, and regulating the spore concentration in the spore suspension to 1-9 multiplied by 10 by using sterile water 7 cfu/mL to obtain the bacterial liquid.
6. The method of claim 5, wherein the activation medium is: 10-15 g of cellobiose, 5-10 g of peptone, 3-5 g of yeast powder, 0.3-1 g of urea and water to 1000mL; ph=6.5 to 7.0.
7. The method according to claim 6, wherein the inoculation mode of the bacterial liquid is spraying inoculation, and the spraying amount of the bacterial liquid is 300-800 kg/hm 2 。
8. The method according to claim 7, wherein the corn stalks are crushed to have a grain size of 1-2 cm and the returning amount of the corn stalks is 2000-3500 kg/hm 2 。
9. The method of claim 8, wherein the corn stover is buried to a depth of 10 to 15cm in the earth.
10. A method according to claim 3, characterized in that the urea is applied in an amount of 200-400 kg/hm 2 The method comprises the steps of carrying out a first treatment on the surface of the The water quantity is 150 to 250t/hm 2 。
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