CN110628675A - Straw field-returning decomposition agent and preparation method thereof - Google Patents
Straw field-returning decomposition agent and preparation method thereof Download PDFInfo
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Classifications
-
- C—CHEMISTRY; METALLURGY
- C05—FERTILISERS; MANUFACTURE THEREOF
- C05F—ORGANIC FERTILISERS NOT COVERED BY SUBCLASSES C05B, C05C, e.g. FERTILISERS FROM WASTE OR REFUSE
- C05F11/00—Other organic fertilisers
- C05F11/08—Organic fertilisers containing added bacterial cultures, mycelia or the like
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/14—Fungi; Culture media therefor
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/14—Fungi; Culture media therefor
- C12N1/16—Yeasts; Culture media therefor
- C12N1/18—Baker's yeast; Brewer's yeast
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
-
- 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
Abstract
A straw field-returning decomposition agent and a preparation method thereof, belonging to the technical field of straw decomposition agents. The decomposing inoculant provided by the invention has the characteristics of high decomposing efficiency and short degrading time, and particularly has a good decomposing effect at normal temperature and even low temperature. The straw field-returning-in-place decomposition agent is prepared by respectively fermenting bacillus amyloliquefaciens, aspergillus niger, trichoderma longibrachiatum and saccharomyces cerevisiae and mixing obtained fermentation liquids, wherein the colony forming unit number ratio of the bacillus amyloliquefaciens fermentation liquid, the aspergillus niger fermentation liquid, the trichoderma longibrachiatum fermentation liquid and the saccharomyces cerevisiae fermentation liquid is 2: 3: 3: 1. the straw field-in-place decomposition agent obtained by the preparation method can promote the straw to be quickly decomposed and returned to the field, supplement organic matters of soil, fully release and utilize the nitrogen, phosphorus and potassium nutrients in the soil and improve the microbial environment of the soil.
Description
Technical Field
The invention belongs to the technical field of straw decomposition agents; in particular to a straw field-returning decomposition agent and a preparation method thereof.
Background
Straw returning is a method for applying straw which is a crop byproduct into soil directly or after being piled up and decomposed. The straws contain a large amount of fresh organic materials, and after the straws return to farmlands, the straws can be converted into organic matters and quick-acting nutrients through decomposition for a period of time, so that the physical and chemical properties of soil can be improved, and certain nutrients can be supplied. The straw returning can promote agriculture water conservation, cost saving, yield increase and efficiency increase, and is fully paid attention to environmental protection and agricultural sustainable development.
The straw returning has a plurality of advantages. The straw returning can increase the fresh organic matters of the soil and improve the soil fertility; the crop straw mainly comprises cellulose, hemicellulose, lignin in a certain proportion, protein and carbohydrate, and the substances are converted into an important composition of soil, namely organic matter, through fermentation and decomposition. Organic matter is an important measure of soil fertility because it is not only a source of major and minor plant nutrient elements, but also plays an important role in preventing soil erosion, increasing water permeability, and improving water utilization. That is, the higher the organic matter content of the soil is, the more fertile the soil is, the better the tilth is, and the more durable the high yield performance is. The straw returning is the most effective measure for increasing the organic matters in the soil. Data published by the general bureau of agricultural cultivation in Heilongjiang show that the continuous returning of crop straws to the field for a long time leads the organic matter of the soil to have a tendency of gradually rising. Particularly, after the wheat straws are returned to the field, the number of bacteria in the soil is increased by 16 times, the number of fiber decomposing bacteria is increased by 8.5 times, and the number of actinomycetes is increased by 3.6 times. The microbial quantity is increased, the activity is enhanced, the decomposition and transformation of soil organic matters are accelerated, and the soil fertilizer supply capability is enhanced.
The existing decomposing inoculant has the problem of too long fermentation time. Under natural conditions, due to the stability of the straw structure, the decomposition and decomposition speed of the straw is quite slow, and the one-year decomposition rate of the straw in the dry land is only less than 50 percent. The composting straw returning process includes collecting straw, applying decomposition agent, fermenting for a while to soften and decompose the straw into fertilizer, and returning the fertilizer to farmland. The decomposition method consumes too much manpower and is not suitable for farmlands operating in large scale. More and more people now pay attention to the technology of straw on-site decomposition and returning to the field, namely, the straw decomposition agent is directly sprayed in the field harvested by the combine harvester to quickly decompose the straw. However, the biggest problem facing field returning in situ is the survival of microorganisms in the straw decomposition agent. Microorganisms exposed in farmlands are inactivated or even killed due to temperature and nutrition problems, and the purpose of returning straws to the field cannot be achieved. The problem of the proportion of low-temperature-resistant microorganism seed selection and other auxiliary bottom materials is the biggest problem in straw decomposition and returning to the field.
Chinese patent CN101139561 discloses a decomposing agent for decomposing straw, which comprises 20% of bacillus amyloliquefaciens, 10% of bacillus licheniformis, 20% of aspergillus flavus, 20% of hair shell, 20% of Umbelopsis pilifera and 10% of yeast, and the process comprises the following steps: respectively purifying, rejuvenating, propagating and culturing the beneficial decomposing bacteria; inoculating each single cultured strain into a sterilized solid culture medium in proportion, fermenting for 4-5 days at the temperature of 25-50 ℃, and airing for later use; then, airing each strain until the moisture content reaches 25%, and preparing the aired single solid strain into the solid decomposing agent according to the formula, uniformly stirring and packaging. Although the decomposing efficiency is high by using the decomposing inoculant, the bacterial strains need to be subjected to rejuvenation culture by using ultraviolet radiation, the preparation process is complex, the cost is too high, and the method is not suitable for commercialization.
Chinese patent CN101306961 discloses a production method and application of a garden plant waste composting microbial inoculum, which comprises inoculating 10-20 parts by mass of each of bacillus amyloliquefaciens, trichoderma viride, lactobacillus plantarum, paracoccus denitrificans, aspergillus oryzae and saccharomyces cerevisiae into a solid fermentation culture medium, and culturing and fermenting for about 3 weeks at 25-30 ℃ in dark and sealed environment; adding 250 parts by mass of sawdust after one week, uniformly stirring, and continuously culturing and fermenting; after two weeks, the hay is placed to absorb excessive water, the hay is removed, and the powder microbial inoculum is obtained after drying in the shade. However, the period of promoting the straw to be completely decomposed by using the microbial inoculum is too long, about one month, and the microbial inoculum is not suitable for being applied to a multi-mature farming area with tight stubble openings.
Disclosure of Invention
The invention aims to provide a preparation method of straw field-returning decomposition agent, and the prepared decomposition agent has the characteristics of high decomposition efficiency and short degradation time, and particularly has good decomposition effect at normal temperature and even low temperature.
In order to solve the technical problems, the straw field-returning-in-place decomposition agent is prepared by respectively fermenting bacillus amyloliquefaciens, aspergillus niger, trichoderma longibrachiatum and saccharomyces cerevisiae and mixing the obtained fermentation liquids, wherein the colony forming unit number ratio of the bacillus amyloliquefaciens fermentation liquid, the aspergillus niger fermentation liquid, the trichoderma longibrachiatum fermentation liquid and the saccharomyces cerevisiae fermentation liquid is 2: 3: 3: 1; the method is specifically completed by the following steps:
respectively starting bacillus amyloliquefaciens, aspergillus niger, trichoderma longibrachiatum and saccharomyces cerevisiae from a test tube inclined plane, performing activated culture and shake flask culture, expanding the culture to a seeding tank for culture, and fermenting in a fermentation tank to respectively obtain bacillus amyloliquefaciens fermentation liquor, aspergillus niger fermentation liquor, trichoderma longibrachiatum fermentation liquor and saccharomyces cerevisiae fermentation liquor;
step two, uniformly mixing the bacillus amyloliquefaciens agent fermentation liquor, the aspergillus niger agent fermentation liquor, the trichoderma longibrachiatum agent fermentation liquor and the saccharomyces cerevisiae fermentation liquor obtained in the step one to obtain the straw field returning and decomposing inoculant.
Further defined, the activation culture in the first step is performed as follows: inoculating the strain in an LB slant culture medium, controlling the temperature to be 35-37 ℃, and culturing for 1-2 days to finish slant culture of the strain in the test tube to obtain an activated strain; wherein the LB slant culture medium is prepared by adding peptone, yeast extract, sodium chloride and agar into water and then uniformly mixing, the pH value is 7.2, and each liter of water contains 9-11 g of peptone, 4-6 g of yeast extract, 9-11 g of sodium chloride and 14-16 g of agar;
further limiting, in the first step, the shake flask activation culture is to inoculate an activated strain obtained by slant culture of a test tube into an LB liquid culture medium, and culture the activated strain for 17 to 19 hours at 35 to 37 ℃ and 160 to 200rpm/min until the number of effective viable bacteria contained in the activated strain reaches 1 to 2 hundred million/ml, so as to complete shake flask activation culture and obtain a culture bacterial liquid; wherein, the LB liquid culture medium is prepared by adding peptone, yeast extract and sodium chloride into water and then uniformly mixing, the pH value is 7.2, and each liter of water contains 9-11 g of peptone, 4-6 g of yeast extract and 9-11 g of sodium chloride.
Further limiting, in the first step, the seeding tank culture is that culture bacterial liquid obtained by the shake flask activation culture is inoculated into a seed culture medium according to the inoculation amount of 1-2%, and then is cultured for 23-25 h under the conditions of 35-37 ℃ and 180-220 rpm/min until the number of effective viable bacteria contained in the culture bacterial liquid reaches 1-2 hundred million/ml, so that the seeding tank fermentation is completed, and seed culture liquid is obtained; wherein, the seed culture medium is prepared by adding peptone, yeast extract and sodium chloride into water and then uniformly mixing, the pH value is 7.2, and each liter of water contains 9-11 g of peptone, 4-6 g of yeast extract and 9-11 g of sodium chloride.
Further limiting, in the fermentation tank fermentation in the first step, the seed culture solution obtained by seed tank culture is inoculated into a fermentation culture medium according to 1-2% of the inoculation amount, and the seed culture solution is cultured for 35-37 h under the conditions of 35-37 ℃ and 180-220 rpm/min until the effective viable count contained in the seed culture solution reaches 2-4 hundred million/ml, so that fermentation tank fermentation is completed, and fermentation broth is obtained; the fermentation medium is prepared by adding corn flour, soybean meal powder, glucose, potassium phosphate, manganese sulfate and calcium carbonate into water and uniformly mixing, wherein each liter of water contains 19-21 g of corn flour, 29-31 g of soybean meal powder, 4-6 g of glucose, 0.9-1.1 g of potassium phosphate, 0.19-0.21 g of manganese sulfate and 0.29-0.31 g of calcium carbonate.
The invention has the beneficial effects that:
(1) the straw field-returning decomposition agent obtained by the preparation method has the advantages of high decomposition efficiency, short degradation time and the like, and particularly has good decomposition effect at normal temperature and even low temperature;
(2) the preparation method has simple process and low preparation cost, and is very suitable for popularization and production;
(3) the straw field-in-place decomposition agent obtained by the preparation method can quickly decompose common crop straws such as wheat straws, rice straws, cereal straws, sweet potato vines, broad bean straws and the like and domestic garbage with high content of fibrous substances on the spot (in a farmland), thereby preparing an organic fertilizer;
(4) the straw field-in-place decomposition agent obtained by the preparation method can promote the straw to be quickly decomposed and returned to the field, supplement organic matters of soil, fully release and utilize the nitrogen, phosphorus and potassium nutrients in the soil and improve the microbial environment of the soil.
Detailed Description
Example 1 in the present embodiment, the straw field-returning-in-place decomposition agent is obtained by respectively fermenting bacillus amyloliquefaciens, aspergillus niger, trichoderma longibrachiatum and saccharomyces cerevisiae, and mixing the obtained fermentation liquids, wherein the colony forming unit number ratio of the bacillus amyloliquefaciens fermentation liquid, the aspergillus niger fermentation liquid, the trichoderma longibrachiatum fermentation liquid and the saccharomyces cerevisiae fermentation liquid is 2: 3: 3: 1; the method is specifically completed by the following steps:
respectively starting bacillus amyloliquefaciens, aspergillus niger, trichoderma longibrachiatum and saccharomyces cerevisiae from a test tube inclined plane, performing activated culture and shake flask culture, expanding the culture to a seeding tank for culture, and fermenting in a fermentation tank to respectively obtain bacillus amyloliquefaciens fermentation liquor, aspergillus niger fermentation liquor, trichoderma longibrachiatum fermentation liquor and saccharomyces cerevisiae fermentation liquor;
step two, uniformly mixing the bacillus amyloliquefaciens fermentation broth obtained in the step one, the aspergillus niger fermentation broth, the trichoderma longibrachiatum fermentation broth and the saccharomyces cerevisiae fermentation broth to obtain a straw field returning and decomposing inoculant;
wherein, the bacillus amyloliquefaciens fermentation liquid in the step one is obtained through the following steps: inoculating bacillus amyloliquefaciens into an LB slant culture medium A, controlling the temperature to be 35 ℃, and culturing for 2 days to finish slant culture of test tube strains to obtain activated strains A; the preparation method of the LB slant culture medium A comprises the following steps: adding peptone, yeast extract, sodium chloride and agar into water, and mixing uniformly to obtain the final product, wherein the pH value is 7.2, and each liter of water contains 9g of peptone, 4g of yeast extract, 9g of sodium chloride and 14g of agar; inoculating the activated strain A into an LB liquid culture medium A, culturing for 19h at 35 ℃ at 200rpm/min until the number of effective viable bacteria contained in the strain reaches 2 hundred million/ml, and completing shake flask activation culture to obtain a culture bacterial liquid A; the preparation method of the LB liquid culture medium A comprises the following steps: adding peptone, yeast extract and sodium chloride into water, and mixing uniformly to obtain the final product, wherein the pH value is 7.2, and each liter of water contains 9g of peptone, 4g of yeast extract and 9g of sodium chloride; inoculating the culture bacterial liquid A into a seed culture medium A according to the inoculation amount of 1%, culturing for 25h at 35 ℃ and 220rpm/min until the number of effective viable bacteria contained therein reaches 2 hundred million/ml, and completing fermentation in a seed tank to obtain a seed culture liquid A; the preparation method of the seed culture medium A comprises the following steps: adding peptone, yeast extract and sodium chloride into water, and mixing uniformly to obtain the final product, wherein the pH value is 7.2, and each liter of water contains 9g of peptone, 4g of yeast extract and 9g of sodium chloride; inoculating the seed culture solution A into a fermentation culture medium A according to the inoculation amount of 1%, culturing for 37h at 35 ℃ and 220rpm/min until the effective viable count contained in the seed culture solution A reaches 4 hundred million/ml, and completing fermentation in a fermentation tank to obtain a bacillus amyloliquefaciens agent fermentation liquid; the preparation method of the fermentation medium A comprises the following steps: adding corn flour, soybean meal powder, glucose, potassium phosphate, manganese sulfate and calcium carbonate into water, and uniformly mixing to obtain the corn flour, the soybean meal powder, the potassium phosphate, the manganese sulfate and the calcium carbonate, wherein each liter of water contains 19g of corn flour, 29g of soybean meal powder, 4g of glucose, 0.9g of potassium phosphate, 0.19g of manganese sulfate and 0.29g of calcium carbonate;
the aspergillus niger microbial inoculum fermentation liquor is obtained by the following steps: inoculating aspergillus niger into an LB slant culture medium B, controlling the temperature to be 35 ℃, and culturing for 2 days to finish slant culture of test tube strains to obtain activated strains B; the preparation method of the LB slant culture medium B comprises the following steps: adding peptone, yeast extract, sodium chloride and agar into water, and mixing uniformly to obtain the final product, wherein the pH value is 7.2, and each liter of water contains 9g of peptone, 4g of yeast extract, 9g of sodium chloride and 14g of agar; inoculating the activated strain B into an LB liquid culture medium B, culturing for 19h at 35 ℃ at 200rpm/min until the number of effective viable bacteria contained in the activated strain B reaches 1 hundred million/ml, and completing shake flask activation culture to obtain a culture bacterial liquid B; the preparation method of the LB liquid culture medium B comprises the following steps: adding peptone, yeast extract and sodium chloride into water, and mixing uniformly to obtain the final product, wherein the pH value is 7.2, and each liter of water contains 9g of peptone, 4g of yeast extract and 9g of sodium chloride; inoculating the culture bacterial liquid B into a seed culture medium B according to the inoculation amount of 1%, culturing for 25h at 35 ℃ and 220rpm/min until the number of effective viable bacteria contained in the culture bacterial liquid B reaches 1 hundred million/ml, and completing fermentation in a seed tank to obtain a seed culture liquid B; the preparation method of the seed culture medium B comprises the following steps: adding peptone, yeast extract and sodium chloride into water, and mixing uniformly to obtain the final product, wherein the pH value is 7.2, and each liter of water contains 9g of peptone, 4g of yeast extract and 9g of sodium chloride; inoculating the seed culture solution B into a fermentation culture medium B according to the inoculation amount of 1%, culturing for 37h at 35 ℃ and 220rpm/min until the effective viable count contained in the seed culture solution B reaches 4 hundred million/ml, and completing fermentation in a fermentation tank to obtain Aspergillus niger microbial inoculum fermentation liquor; the preparation method of the fermentation medium B comprises the following steps: adding corn flour, soybean meal powder, glucose, potassium phosphate, manganese sulfate and calcium carbonate into water, and uniformly mixing to obtain the corn flour, the soybean meal powder, the potassium phosphate, the manganese sulfate and the calcium carbonate, wherein each liter of water contains 19g of corn flour, 29g of soybean meal powder, 4g of glucose, 0.9g of potassium phosphate, 0.19g of manganese sulfate and 0.29g of calcium carbonate;
the trichoderma longibrachiatum microbial inoculum fermentation liquor is obtained by the following steps: inoculating trichoderma longibrachiatum in an LB slant culture medium C, controlling the temperature to be 35 ℃, and culturing for 2 days to finish slant culture of test tube strains to obtain activated strains C; the preparation method of the LB slant culture medium C comprises the following steps: adding peptone, yeast extract, sodium chloride and agar into water, and mixing uniformly to obtain the final product, wherein the pH value is 7.2, and each liter of water contains 9g of peptone, 4g of yeast extract, 9g of sodium chloride and 14g of agar; inoculating the activated strain C into an LB liquid culture medium C, culturing for 19h at 35 ℃ at 200rpm/min until the number of effective viable bacteria contained in the activated strain C reaches 1 hundred million/ml, and completing shake flask activation culture to obtain a culture bacterial liquid C; the preparation method of the LB liquid culture medium C comprises the following steps: adding peptone, yeast extract and sodium chloride into water, and mixing uniformly to obtain the final product, wherein the pH value is 7.2, and each liter of water contains 9g of peptone, 4g of yeast extract and 9g of sodium chloride; inoculating the culture bacterial liquid C into a seed culture medium C according to the inoculation amount of 1%, culturing for 25h at 35 ℃ and 220rpm/min until the number of effective viable bacteria contained in the culture bacterial liquid C reaches 1 hundred million/ml, and completing fermentation in a seed tank to obtain a seed culture liquid C; the preparation method of the seed culture medium C comprises the following steps: adding peptone, yeast extract and sodium chloride into water, and mixing uniformly to obtain the final product, wherein the pH value is 7.2, and each liter of water contains 9g of peptone, 4g of yeast extract and 9g of sodium chloride; inoculating the seed culture solution C into a fermentation culture medium C according to the inoculation amount of 1%, culturing for 37h at 35 ℃ and 220rpm/min until the effective viable count contained in the seed culture solution C reaches 2 hundred million/ml, and completing fermentation in a fermentation tank to obtain trichoderma longibrachiatum microbial inoculum fermentation liquor; the preparation method of the fermentation medium C comprises the following steps: adding corn flour, soybean meal powder, glucose, potassium phosphate, manganese sulfate and calcium carbonate into water, and uniformly mixing to obtain the corn flour, the soybean meal powder, the potassium phosphate, the manganese sulfate and the calcium carbonate, wherein each liter of water contains 19g of corn flour, 29g of soybean meal powder, 4g of glucose, 0.9g of potassium phosphate, 0.19g of manganese sulfate and 0.29g of calcium carbonate;
the saccharomyces cerevisiae fermentation liquor is obtained by the following steps: inoculating Saccharomyces cerevisiae into PDA slant culture medium, controlling temperature at 27 deg.C, and culturing for 3 days to complete slant culture of test tube strain to obtain activated strain D; the preparation method of the DA slant culture medium comprises the following steps: adding potato and sucrose into water, wherein each liter of water contains 190g potato and 19g sucrose; inoculating the activated strain D into a PDA liquid culture medium, culturing for 37h at 27 ℃ and 200rpm/min until the number of effective viable bacteria contained in the activated strain D reaches 1 hundred million/ml, and completing shake flask activation culture to obtain a culture bacterial liquid D; the preparation method of the PDA liquid culture medium comprises the following steps: adding potato and sucrose into water, wherein each liter of water contains 190g potato and 19g sucrose; inoculating the culture bacterial liquid D into a seed culture medium D according to the inoculation amount of 1%, culturing for 37h at 27 ℃ and 220rpm/min until the number of effective viable bacteria contained in the culture bacterial liquid D reaches 2 hundred million/ml, and completing fermentation in a seed tank to obtain a seed culture liquid D; the preparation method of the seed culture medium D comprises the following steps: adding glucose, peptone and yeast extract into water, and mixing well, wherein each liter of water contains 4g of glucose, 4g of peptone and 2g of yeast extract; inoculating the seed culture solution D into a fermentation culture medium D according to the inoculation amount of 4%, culturing for 47h at 27 ℃ under the condition of 220rpm/min until the effective viable count contained in the seed culture solution D reaches 4 hundred million/ml, and completing fermentation in a fermentation tank to obtain saccharomyces cerevisiae fermentation liquor; the preparation method of the fermentation medium D comprises the following steps: adding glucose, yeast extract, peptone and calcium carbonate into water, and mixing well, wherein each liter of water contains 19g of glucose, 9g of yeast extract, 4g of peptone and 9g of calcium carbonate.
Example 2: in this embodiment, the straw field-returning-in-place decomposition agent is obtained by respectively fermenting bacillus amyloliquefaciens, aspergillus niger, trichoderma longibrachiatum and saccharomyces cerevisiae, and mixing the obtained fermentation liquids, wherein the colony forming unit number ratio of the bacillus amyloliquefaciens fermentation liquid, the aspergillus niger fermentation liquid, the trichoderma longibrachiatum fermentation liquid and the saccharomyces cerevisiae fermentation liquid is 2: 3: 3: 1; the method is specifically completed by the following steps:
respectively starting bacillus amyloliquefaciens, aspergillus niger, trichoderma longibrachiatum and saccharomyces cerevisiae from a test tube inclined plane, performing activated culture and shake flask culture, expanding the culture to a seeding tank for culture, and fermenting in a fermentation tank to respectively obtain bacillus amyloliquefaciens fermentation liquor, aspergillus niger fermentation liquor, trichoderma longibrachiatum fermentation liquor and saccharomyces cerevisiae fermentation liquor;
step two, uniformly mixing the bacillus amyloliquefaciens fermentation broth obtained in the step one, the aspergillus niger fermentation broth, the trichoderma longibrachiatum fermentation broth and the saccharomyces cerevisiae fermentation broth to obtain a straw field returning and decomposing inoculant;
wherein, the bacillus amyloliquefaciens fermentation liquid in the step one is obtained through the following steps: inoculating bacillus amyloliquefaciens into an LB slant culture medium A, controlling the temperature to be 36 ℃, and culturing for 1 day to finish slant culture of test tube strains to obtain activated strains A; the preparation method of the LB slant culture medium A comprises the following steps: adding peptone, yeast extract, sodium chloride and agar into water, and mixing uniformly to obtain the final product, wherein the pH value is 7.2, and each liter of water contains 10g of peptone, 5g of yeast extract, 10g of sodium chloride and 15g of agar; inoculating the activated strain A into an LB liquid culture medium A, culturing for 18h at 36 ℃ and 180rpm/min until the number of effective viable bacteria contained in the activated strain A reaches 1.5 hundred million/ml, and completing shake flask activation culture to obtain a culture bacterial liquid A; the preparation method of the LB liquid culture medium A comprises the following steps: adding peptone, yeast extract and sodium chloride into water, and mixing uniformly to obtain the final product, wherein the pH value is 7.2, and each liter of water contains 10g of peptone, 5g of yeast extract and 10g of sodium chloride; inoculating the culture bacterial liquid A into a seed culture medium A according to the inoculation amount of 1.5%, culturing for 24h at 36 ℃ under the condition of 180pm/min until the number of effective viable bacteria contained in the culture bacterial liquid A reaches 1.5 hundred million/ml, and completing fermentation in a seed tank to obtain a seed culture liquid A; the preparation method of the seed culture medium A comprises the following steps: adding peptone, yeast extract and sodium chloride into water, and mixing uniformly to obtain the final product, wherein the pH value is 7.2, and each liter of water contains 10g of peptone, 5g of yeast extract and 10g of sodium chloride; inoculating the seed culture solution A into a fermentation culture medium A according to the inoculation amount of 1.5%, culturing for 36h at 36 ℃ under the condition of 200rpm/min until the effective viable count contained in the seed culture solution A reaches 3 hundred million/ml, and completing fermentation in a fermentation tank to obtain a bacillus amyloliquefaciens agent fermentation liquid; the preparation method of the fermentation medium A comprises the following steps: adding corn flour, soybean meal powder, glucose, potassium phosphate, manganese sulfate and calcium carbonate into water, and uniformly mixing to obtain the corn flour, the soybean meal powder, the glucose powder, the potassium phosphate, the manganese sulfate and the calcium carbonate, wherein each liter of water contains 20g of corn flour, 30g of soybean meal powder, 5g of glucose, 1.0g of potassium phosphate, 0.2g of manganese sulfate and 0.3 g of calcium carbonate;
the aspergillus niger microbial inoculum fermentation liquor is obtained by the following steps: inoculating aspergillus niger into an LB slant culture medium B, controlling the temperature to be 36 ℃, and culturing for 1 day to finish slant culture of test tube strains to obtain activated strains B; the preparation method of the LB slant culture medium B comprises the following steps: adding peptone, yeast extract, sodium chloride and agar into water, and mixing uniformly to obtain the final product, wherein the pH value is 7.2, and each liter of water contains 10g of peptone, 5g of yeast extract, 10g of sodium chloride and 15g of agar; inoculating the activated strain B into an LB liquid culture medium B, culturing for 18h at 36 ℃ and 180rpm/min until the number of effective viable bacteria contained in the activated strain B reaches 1.5 hundred million/ml, and completing shake flask activation culture to obtain a culture bacterial liquid B; the preparation method of the LB liquid culture medium B comprises the following steps: adding peptone, yeast extract and sodium chloride into water, and mixing uniformly to obtain the final product, wherein the pH value is 7.2, and each liter of water contains 10g of peptone, 5g of yeast extract and 10g of sodium chloride; inoculating the culture bacterial liquid B into a seed culture medium B according to the inoculation amount of 1.5%, culturing for 24h at 35 ℃ under the condition of 200rpm/min until the number of effective viable bacteria contained in the culture bacterial liquid B reaches 1.5 hundred million/ml, and completing fermentation in a seed tank to obtain a seed culture liquid B; the preparation method of the seed culture medium B comprises the following steps: adding peptone, yeast extract and sodium chloride into water, and mixing uniformly to obtain the final product, wherein the pH value is 7.2, and each liter of water contains 10g of peptone, 5g of yeast extract and 10g of sodium chloride; inoculating the seed culture solution B into a fermentation culture medium B according to the inoculation amount of 1.5%, culturing for 36h at 36 ℃ at 200rpm/min until the effective viable count contained in the seed culture solution B reaches 3 hundred million/ml, and completing fermentation in a fermentation tank to obtain Aspergillus niger agent fermentation liquor; the preparation method of the fermentation medium B comprises the following steps: adding corn flour, soybean meal powder, glucose, potassium phosphate, manganese sulfate and calcium carbonate into water, and uniformly mixing to obtain the corn flour, the soybean meal powder, the glucose powder, the potassium phosphate, the manganese sulfate and the calcium carbonate, wherein each liter of water contains 20g of corn flour, 30g of soybean meal powder, 5g of glucose, 1.0g of potassium phosphate, 0.2g of manganese sulfate and 0.3 g of calcium carbonate;
the long-branch wood bacterium agent fermentation liquor is obtained by the following steps: inoculating trichoderma longibrachiatum in an LB slant culture medium C, controlling the temperature to be 36 ℃, and culturing for 1 day to finish slant culture of test tube strains to obtain activated strains C; the preparation method of the LB slant culture medium C comprises the following steps: adding peptone, yeast extract, sodium chloride and agar into water, and mixing uniformly to obtain the final product, wherein the pH value is 7.2, and each liter of water contains 10g of peptone, 5g of yeast extract, 10g of sodium chloride and 15g of agar; inoculating the activated strain C into an LB liquid culture medium C, culturing for 18h at 36 ℃ and 180rpm/min until the number of effective viable bacteria contained in the activated strain C reaches 1.5 hundred million/ml, and completing shake flask activation culture to obtain a culture bacterial liquid C; the preparation method of the LB liquid culture medium C comprises the following steps: adding peptone, yeast extract and sodium chloride into water, and mixing uniformly to obtain the final product, wherein the pH value is 7.2, and each liter of water contains 10g of peptone, 5g of yeast extract and 10g of sodium chloride; inoculating the culture bacterial liquid C into a seed culture medium C according to the inoculation amount of 1.5%, culturing for 24h at 36 ℃ under the condition of 200rpm/min until the number of effective viable bacteria contained in the culture bacterial liquid C reaches 1.5 hundred million/ml, and completing fermentation in a seed tank to obtain a seed culture liquid C; the preparation method of the seed culture medium C comprises the following steps: adding peptone, yeast extract and sodium chloride into water, and mixing uniformly to obtain the final product, wherein the pH value is 7.2, and each liter of water contains 10g of peptone, 5g of yeast extract and 10g of sodium chloride; inoculating the seed culture solution C into a fermentation culture medium C according to the inoculation amount of 1.5%, culturing for 36h at 36 ℃ at 200rpm/min until the effective viable count contained in the seed culture solution C reaches 3 hundred million/ml, and completing fermentation in a fermentation tank to obtain trichoderma longibrachiatum inoculum fermentation liquid; the preparation method of the fermentation medium C comprises the following steps: adding corn flour, soybean meal powder, glucose, potassium phosphate, manganese sulfate and calcium carbonate into water, and uniformly mixing to obtain the corn flour, the soybean meal powder, the glucose powder, the potassium phosphate, the manganese sulfate and the calcium carbonate, wherein each liter of water contains 20g of corn flour, 30g of soybean meal powder, 5g of glucose, 1.0g of potassium phosphate, 0.2g of manganese sulfate and 0.3 g of calcium carbonate;
the saccharomyces cerevisiae fermentation liquor is obtained by the following steps: inoculating saccharomyces cerevisiae in a PDA slant culture medium, controlling the temperature at 28 ℃, and culturing for 2 days to complete slant culture of test tube strains to obtain activated strains D; the preparation method of the DA slant culture medium comprises the following steps: adding potato and sucrose into water, wherein each liter of water contains 200g potato and 20g sucrose; inoculating the activated strain D into a PDA liquid culture medium, culturing for 36h at 28 ℃ and 180rpm/min until the number of effective viable bacteria contained in the activated strain D reaches 1.5 hundred million/ml, and completing shake flask activation culture to obtain a culture bacterial liquid D; the preparation method of the PDA liquid culture medium comprises the following steps: adding potato and sucrose into water, wherein each liter of water contains 200g potato and 20g sucrose; inoculating the culture bacterial liquid D into a seed culture medium D according to the inoculation amount of 1.5%, culturing for 36h at 28 ℃ under the condition of 200rpm/min until the number of effective viable bacteria contained in the culture bacterial liquid D reaches 1.5 hundred million/ml, and completing fermentation in a seed tank to obtain a seed culture liquid D; the preparation method of the seed culture medium D comprises the following steps: adding glucose, peptone and yeast extract into water, and mixing well, wherein each liter of water contains 5g of glucose, 5g of peptone and 3g of yeast extract; inoculating the seed culture solution D into a fermentation culture medium D according to the inoculation amount of 5%, culturing for 48h at 28 ℃ under the condition of 200rpm/min until the effective viable count contained in the seed culture solution D reaches 3 hundred million/ml, and completing fermentation in a fermentation tank to obtain saccharomyces cerevisiae fermentation liquor; the preparation method of the fermentation medium D comprises the following steps: adding glucose, yeast extract, peptone and calcium carbonate into water, and mixing well, wherein each liter of water contains 20g of glucose, 10g of yeast extract, 5g of peptone and 10g of calcium carbonate.
Example 3: in this embodiment, the straw field-returning-in-place decomposition agent is obtained by respectively fermenting bacillus amyloliquefaciens, aspergillus niger, trichoderma longibrachiatum and saccharomyces cerevisiae, and mixing the obtained fermentation liquids, wherein the colony forming unit number ratio of the bacillus amyloliquefaciens fermentation liquid, the aspergillus niger fermentation liquid, the trichoderma longibrachiatum fermentation liquid and the saccharomyces cerevisiae fermentation liquid is 2: 3: 3: 1; the method is specifically completed by the following steps:
respectively starting bacillus amyloliquefaciens, aspergillus niger, trichoderma longibrachiatum and saccharomyces cerevisiae from a test tube inclined plane, performing activated culture and shake flask culture, expanding the culture to a seeding tank for culture, and fermenting in a fermentation tank to respectively obtain bacillus amyloliquefaciens fermentation liquor, aspergillus niger fermentation liquor, trichoderma longibrachiatum fermentation liquor and saccharomyces cerevisiae fermentation liquor;
step two, uniformly mixing the bacillus amyloliquefaciens fermentation broth obtained in the step one, the aspergillus niger fermentation broth, the trichoderma longibrachiatum fermentation broth and the saccharomyces cerevisiae fermentation broth to obtain a straw field returning and decomposing inoculant;
wherein, the bacillus amyloliquefaciens fermentation liquid in the step one is obtained through the following steps: inoculating bacillus amyloliquefaciens into an LB slant culture medium A, controlling the temperature to be 37 ℃, and culturing for 1 day to finish slant culture of test tube strains to obtain activated strains A; the preparation method of the LB slant culture medium A comprises the following steps: adding peptone, yeast extract, sodium chloride and agar into water, and mixing uniformly to obtain the final product, wherein the pH value is 7.2, and each liter of water contains 11g of peptone, 6g of yeast extract, 11g of sodium chloride and 16g of agar; inoculating the activated strain A into an LB liquid culture medium A, culturing for 17h at 37 ℃ and 160rpm/min until the number of effective viable bacteria contained in the activated strain A reaches 1 hundred million/ml, and completing shake flask activation culture to obtain a culture bacterial liquid A; the preparation method of the LB liquid culture medium A comprises the following steps: adding peptone, yeast extract and sodium chloride into water, and mixing uniformly to obtain the final product, wherein the pH value is 7.2, and each liter of water contains 11g of peptone, 6g of yeast extract and 16g of sodium chloride; inoculating the culture bacterial liquid A into a seed culture medium A according to the inoculation amount of 2%, culturing for 23h at 37 ℃ under the condition of 180pm/min until the number of effective viable bacteria contained in the culture bacterial liquid reaches 1 hundred million/ml, and completing fermentation in a seed tank to obtain a seed culture liquid A; the preparation method of the seed culture medium A comprises the following steps: adding peptone, yeast extract and sodium chloride into water, and mixing uniformly to obtain the final product, wherein the pH value is 7.2, and each liter of water contains 11g of peptone, 6g of yeast extract and 11g of sodium chloride; inoculating the seed culture solution A into a fermentation culture medium A according to the inoculation amount of 2%, culturing for 35h at 37 ℃ under the condition of 180rpm/min until the effective viable count contained in the seed culture solution A reaches 2 hundred million/ml, and completing fermentation in a fermentation tank to obtain a bacillus amyloliquefaciens agent fermentation liquid; the preparation method of the fermentation medium A comprises the following steps: adding corn flour, soybean meal powder, glucose, potassium phosphate, manganese sulfate and calcium carbonate into water, and uniformly mixing to obtain the corn flour, the soybean meal powder, the glucose powder, the potassium phosphate, the manganese sulfate and the calcium carbonate, wherein each liter of water contains 21g of corn flour, 31g of soybean meal powder, 6g of glucose, 1.1g of potassium phosphate, 0.21g of manganese sulfate and 0.31g of calcium carbonate;
the aspergillus niger microbial inoculum fermentation liquor is obtained by the following steps: inoculating aspergillus niger into an LB slant culture medium B, controlling the temperature to be 37 ℃, and culturing for 1 day to finish slant culture of test tube strains to obtain activated strains B; the preparation method of the LB slant culture medium B comprises the following steps: adding peptone, yeast extract, sodium chloride and agar into water, and mixing uniformly to obtain the final product, wherein the pH value is 7.2, and each liter of water contains 11g of peptone, 6g of yeast extract, 11g of sodium chloride and 15g of agar; inoculating the activated strain B into an LB liquid culture medium B, culturing for 17h at 37 ℃ and 160rpm/min until the number of effective viable bacteria contained in the activated strain B reaches 1 hundred million/ml, and completing shake flask activation culture to obtain a culture bacterial liquid B; the preparation method of the LB liquid culture medium B comprises the following steps: adding peptone, yeast extract and sodium chloride into water, and mixing uniformly to obtain the final product, wherein the pH value is 7.2, and each liter of water contains 11g of peptone, 6g of yeast extract and 11g of sodium chloride; inoculating the culture bacterial liquid B into a seed culture medium B according to the inoculation amount of 2%, culturing for 23h at 37 ℃ under the condition of 180rpm/min until the number of effective viable bacteria contained in the culture bacterial liquid B reaches 4 hundred million/ml, and completing fermentation in a seed tank to obtain a seed culture liquid B; the preparation method of the seed culture medium B comprises the following steps: adding peptone, yeast extract and sodium chloride into water, and mixing uniformly to obtain the final product, wherein the pH value is 7.2, and each liter of water contains 11g of peptone, 6g of yeast extract and 11g of sodium chloride; inoculating the seed culture solution B into a fermentation culture medium B according to the inoculation amount of 2%, culturing for 35h at 37 ℃ under the condition of 180rpm/min until the effective viable count contained in the seed culture solution B reaches 2 hundred million/ml, and completing fermentation in a fermentation tank to obtain Aspergillus niger agent fermentation liquor; the preparation method of the fermentation medium B comprises the following steps: adding corn flour, soybean meal powder, glucose, potassium phosphate, manganese sulfate and calcium carbonate into water, and uniformly mixing to obtain the corn flour, the soybean meal powder, the glucose powder, the potassium phosphate, the manganese sulfate and the calcium carbonate, wherein each liter of water contains 21g of corn flour, 31g of soybean meal powder, 6g of glucose, 1.1g of potassium phosphate, 0.21g of manganese sulfate and 0.31g of calcium carbonate;
the trichoderma longibrachiatum microbial inoculum fermentation liquor is obtained by the following steps: inoculating trichoderma longibrachiatum in an LB slant culture medium C, controlling the temperature to be 37 ℃, and culturing for 1 day to finish slant culture of test tube strains to obtain activated strains C; the preparation method of the LB slant culture medium C comprises the following steps: adding peptone, yeast extract, sodium chloride and agar into water, and mixing uniformly to obtain the final product, wherein the pH value is 7.2, and each liter of water contains 11g of peptone, 6g of yeast extract, 11g of sodium chloride and 16g of agar; inoculating the activated strain C into an LB liquid culture medium C, culturing for 17h at 37 ℃ and 160rpm/min until the number of effective viable bacteria contained in the activated strain C reaches 1 hundred million/ml, and completing shake flask activation culture to obtain a culture bacterial liquid C; the preparation method of the LB liquid culture medium C comprises the following steps: adding peptone, yeast extract and sodium chloride into water, and mixing uniformly to obtain the final product, wherein the pH value is 7.2, and each liter of water contains 11g of peptone, 6g of yeast extract and 11g of sodium chloride; inoculating the culture bacterial liquid C into a seed culture medium C according to the inoculation amount of 2%, culturing for 23h at 37 ℃ under the condition of 180rpm/min until the number of effective viable bacteria contained in the culture bacterial liquid C reaches 1 hundred million/ml, and completing fermentation in a seed tank to obtain a seed culture liquid C; the preparation method of the seed culture medium C comprises the following steps: adding peptone, yeast extract and sodium chloride into water, and mixing uniformly to obtain the final product, wherein the pH value is 7.2, and each liter of water contains 11g of peptone, 6g of yeast extract and 11g of sodium chloride; inoculating the seed culture solution C into a fermentation culture medium C according to the inoculation amount of 2%, culturing for 35h at 37 ℃ under the condition of 180rpm/min until the effective viable count contained in the seed culture solution C reaches 2 hundred million/ml, and completing fermentation in a fermentation tank to obtain trichoderma longibrachiatum microbial inoculum fermentation liquor; the preparation method of the fermentation medium C comprises the following steps: adding corn flour, soybean meal powder, glucose, potassium phosphate, manganese sulfate and calcium carbonate into water, and uniformly mixing to obtain the corn flour, the soybean meal powder, the glucose powder, the potassium phosphate, the manganese sulfate and the calcium carbonate, wherein each liter of water contains 21g of corn flour, 31g of soybean meal powder, 6g of glucose, 1.1g of potassium phosphate, 0.21g of manganese sulfate and 0.31g of calcium carbonate;
the saccharomyces cerevisiae fermentation liquor is obtained by the following steps: inoculating Saccharomyces cerevisiae into PDA slant culture medium, controlling temperature at 29 deg.C, and culturing for 2 days to complete slant culture of test tube strain to obtain activated strain D; the preparation method of the DA slant culture medium comprises the following steps: adding potato and sucrose into water, wherein each liter of water contains 210g potato and 21g sucrose; inoculating the activated strain D into a PDA liquid culture medium, culturing for 35h at 29 ℃ and 160rpm/min until the number of effective viable bacteria contained in the activated strain D reaches 1 hundred million/ml, and completing shake flask activation culture to obtain a culture bacterial liquid D; the preparation method of the PDA liquid culture medium comprises the following steps: adding potato and sucrose into water, wherein each liter of water contains 210g potato and 21g sucrose; inoculating the culture bacterial liquid D into a seed culture medium D according to the inoculation amount of 2%, culturing for 35h at 29 ℃ under the condition of 180rpm/min until the number of effective viable bacteria contained in the culture bacterial liquid D reaches 1 hundred million/ml, and completing fermentation in a seed tank to obtain a seed culture liquid D; the preparation method of the seed culture medium D comprises the following steps: adding glucose, peptone and yeast extract into water, and mixing well, wherein each liter of water contains 6g of glucose, 6g of peptone and 4g of yeast extract; inoculating the seed culture solution D into a fermentation culture medium D according to the inoculation amount of 6%, culturing for 47h at 29 ℃ under the condition of 180rpm/min until the effective viable count contained in the seed culture solution D reaches 2 hundred million/ml, and completing fermentation in a fermentation tank to obtain saccharomyces cerevisiae fermentation liquor; the preparation method of the fermentation medium D comprises the following steps: adding glucose, yeast extract, peptone and calcium carbonate into water, and mixing well, wherein each liter of water contains 21g of glucose, 11g of yeast extract, 6g of peptone and 11g of calcium carbonate.
The using conditions of the straw field-returning decomposition agent are as follows: the optimal inoculation amount in the field is 4 percent, namely 4g of degrading bacteria agent needs to be inoculated to each kilogram of straws; when the microbial inoculum is applied to a field, nitrogen and phosphorus can be applied according to needs, the recommended application amount of nitrogen per kilogram of straw is 12g, the addition amount of phosphorus is 5g, the larger the amount is, the more the straw degradation is facilitated, and therefore the phosphorus addition amount can be determined according to actual conditions.
The application method of the straw field-returning decomposition agent comprises the following steps: while returning the field with the straw, spreading or spraying the straw field-returning decomposition agent; the using amount is 2-3 kilograms per mu, and the straw is spread or sprayed with the straw field returning decomposition agent; meanwhile, nitrogen fertilizer and phosphate fertilizer are applied, and then the straws are turned into the ground, so that the straw compost is also suitable for rotten fermentation under the condition of straw stacking.
Through screening and researching a large number of microbial strains, the following results are finally found: the straw field-in-place returning and decomposing agent obtained by matching aspergillus niger, saccharomyces cerevisiae, trichoderma longibrachiatum and bacillus amyloliquefaciens (the four functional bacteria are combined together to have a synergistic effect) can promote the straws to be rapidly decomposed and returned to the field, supplement soil organic matters, fully release and utilize nitrogen, phosphorus and potassium nutrients in the soil, and improve the microbial environment of the soil.
Aspergillus niger is a strong decomposer of crop straws, has high enzyme activity, and a large number of spores of Aspergillus niger can quickly germinate at the temperature of more than 0 ℃ to generate heat, so that the local ambient temperature around is increased, and necessary temperature conditions are provided for the quick propagation of the Aspergillus niger and other functional bacteria.
The trichoderma longibrachiatum has outstanding cellulase activity and has obvious effect of promoting the decomposition of straws mainly containing cellulose.
The bacillus amyloliquefaciens also has strong decomposition capacity on crop straws and high enzyme activity of 'tripsin', quickly germinates by virtue of the temperature rise effect of aspergillus niger so as to bring the effect of continuous temperature rise on the environment, further improves the enzyme activity and accelerates the decomposition of the straws.
The saccharomyces cerevisiae can decompose and utilize metabolites of the first three functional bacteria, eliminate the growth inhibition of the first three functional bacteria on the metabolites, promote the growth and the propagation of other indigenous microorganisms, accelerate the decomposition speed and improve the soil microbial environment.
The straw field-in-place decomposition agent obtained by the preparation method can be normally used in an environment with the temperature of more than 0 ℃, the temperature of the straw reaches more than 15 ℃ after the straw is decomposed for 3 days in an environment with the temperature of about 4 ℃, the field-in-place straw is completely decomposed in about 18 days, and a good decomposition effect can be achieved in an environment with the temperature of more than 20 ℃ in about 10 days.
Claims (10)
1. The straw field returning-in-place decomposition agent is characterized by being prepared by respectively fermenting bacillus amyloliquefaciens, aspergillus niger, trichoderma longibrachiatum and saccharomyces cerevisiae and mixing obtained fermentation liquids, wherein the colony forming unit number ratio of the bacillus amyloliquefaciens fermentation liquid, the aspergillus niger fermentation liquid, the trichoderma longibrachiatum fermentation liquid and the saccharomyces cerevisiae fermentation liquid is 2: 3: 3: 1.
2. the method for preparing the straw field-returning decomposition agent according to claim 1 is characterized by comprising the following steps:
respectively starting bacillus amyloliquefaciens, aspergillus niger, trichoderma longibrachiatum and saccharomyces cerevisiae from a test tube inclined plane, performing activated culture and shake flask culture, expanding the culture to a seeding tank for culture, and fermenting in a fermentation tank to respectively obtain bacillus amyloliquefaciens fermentation liquor, aspergillus niger fermentation liquor, trichoderma longibrachiatum fermentation liquor and saccharomyces cerevisiae fermentation liquor;
step two, uniformly mixing the bacillus amyloliquefaciens agent fermentation liquor, the aspergillus niger agent fermentation liquor, the trichoderma longibrachiatum agent fermentation liquor and the saccharomyces cerevisiae fermentation liquor obtained in the step one to obtain the straw field returning and decomposing inoculant.
3. The method for preparing straw field-returning decomposition agent according to claim 2, wherein the activation culture in the first step is performed as follows: inoculating the strain in LB slant culture medium at 35-37 deg.c for 1-2 days to complete slant culture of the strain in test tube and obtain the activated strain.
4. The method for preparing the straw field-returning decomposition agent as claimed in claim 3, wherein the LB slant culture medium is prepared by adding peptone, yeast extract, sodium chloride and agar into water and mixing them uniformly, the pH value is 7.2, and each liter of water contains 9-11 g of peptone, 4-6 g of yeast extract, 9-11 g of sodium chloride and 14-16 g of agar.
5. The method for preparing straw field-returning decomposition agent according to claim 2, wherein in the first step of shake flask activation culture, activated strains obtained by slant culture of test tubes are inoculated into LB liquid medium, and cultured for 17-19 h at 35-37 ℃ and 160-200 rpm/min until the number of effective viable bacteria contained therein reaches 1-2 hundred million/ml, and shake flask activation culture is completed to obtain culture bacteria liquid.
6. The method for preparing straw field-returning decomposition agent according to claim 5, wherein the LB liquid medium is prepared by adding peptone, yeast extract and sodium chloride into water and mixing them uniformly, the pH value is 7.2, and each liter of water contains 9-11 g of peptone, 4-6 g of yeast extract and 9-11 g of sodium chloride.
7. The method for preparing straw field-returning decomposition agent according to claim 2, wherein the first step of the seeding tank culture is that the culture bacterial liquid obtained by the shake flask activation culture is inoculated into the seed culture medium according to the inoculation amount of 1-2%, and then cultured for 23-25 h under the conditions of 35-37 ℃ and 180-220 rpm/min until the effective viable count contained therein reaches 1-2 hundred million/ml, and the seeding tank fermentation is completed to obtain the seed culture liquid.
8. The method for preparing straw field-returning decomposition agent according to claim 7, wherein the seed culture medium is prepared by adding peptone, yeast extract and sodium chloride into water and mixing uniformly, the pH value is 7.2, and each liter of water contains 9-11 g of peptone, 4-6 g of yeast extract and 9-11 g of sodium chloride.
9. The method for preparing straw field-returning decomposition agent according to claim 2, wherein in the fermentation tank fermentation in the first step, the seed culture solution obtained by the seed tank culture is inoculated into the fermentation culture medium according to 1% -2% of the inoculum size, and the fermentation is carried out for 35-37 h at 35-37 ℃ and 180-220 rpm/min until the effective viable count contained therein reaches 2-4 hundred million/ml, and the fermentation tank fermentation is completed to obtain the fermentation liquid.
10. The method for preparing the straw field-returning decomposition agent as claimed in claim 9, wherein the fermentation medium is prepared by adding corn flour, soybean meal powder, glucose, potassium phosphate, manganese sulfate and calcium carbonate into water and mixing uniformly, wherein each liter of water contains 19-21 g of corn flour, 29-31 g of soybean meal powder, 4-6 g of glucose, 0.9-1.1 g of potassium phosphate, 0.19-0.21 g of manganese sulfate and 0.29-0.31 g of calcium carbonate.
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Application publication date: 20191231 |