CN114230402B - Efficient decomposed liquid bacterial fertilizer and preparation method and application thereof - Google Patents
Efficient decomposed liquid bacterial fertilizer and preparation method and application thereof Download PDFInfo
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- CN114230402B CN114230402B CN202110877770.4A CN202110877770A CN114230402B CN 114230402 B CN114230402 B CN 114230402B CN 202110877770 A CN202110877770 A CN 202110877770A CN 114230402 B CN114230402 B CN 114230402B
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- C—CHEMISTRY; METALLURGY
- C05—FERTILISERS; MANUFACTURE THEREOF
- C05G—MIXTURES OF FERTILISERS COVERED INDIVIDUALLY BY DIFFERENT SUBCLASSES OF CLASS C05; MIXTURES OF ONE OR MORE FERTILISERS WITH MATERIALS NOT HAVING A SPECIFIC FERTILISING ACTIVITY, e.g. PESTICIDES, SOIL-CONDITIONERS, WETTING AGENTS; FERTILISERS CHARACTERISED BY THEIR FORM
- C05G3/00—Mixtures of one or more fertilisers with additives not having a specially fertilising activity
- C05G3/40—Mixtures of one or more fertilisers with additives not having a specially fertilising activity for affecting fertiliser dosage or release rate; for affecting solubility
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- C—CHEMISTRY; METALLURGY
- C05—FERTILISERS; MANUFACTURE THEREOF
- C05C—NITROGENOUS FERTILISERS
- C05C9/00—Fertilisers containing urea or urea compounds
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- C—CHEMISTRY; METALLURGY
- C05—FERTILISERS; MANUFACTURE THEREOF
- C05G—MIXTURES OF FERTILISERS COVERED INDIVIDUALLY BY DIFFERENT SUBCLASSES OF CLASS C05; MIXTURES OF ONE OR MORE FERTILISERS WITH MATERIALS NOT HAVING A SPECIFIC FERTILISING ACTIVITY, e.g. PESTICIDES, SOIL-CONDITIONERS, WETTING AGENTS; FERTILISERS CHARACTERISED BY THEIR FORM
- C05G5/00—Fertilisers characterised by their form
- C05G5/20—Liquid fertilisers
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- 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
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- 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
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- 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
The invention discloses a high-efficiency decomposed liquid bacterial fertilizer and a preparation method thereof, wherein the bacterial fertilizer comprises bacillus subtilis, streptomyces roseoflavus, trichoderma harzianum, penicillium cruentum, myceliophthora thermophila and pediococcus pentosaceus, and the number of effective viable bacteria is more than or equal to 1 multiplied by 10 9 cfu/mL. The invention has the advantages that the prepared bacterial manure liquid contains sufficient nitrogen source required by the growth and metabolism of microorganisms, and the straw decomposition efficiency can be effectively improved without additionally supplementing the nitrogen source during application; on the other hand, various metabolic pathways of different strains and complementation on different degrading enzyme systems are fused, pediococcus pentosaceus can effectively degrade intermediate metabolites of straw lignocellulose, such as low-molecular oligosaccharides, lactic acid, ethanol and the like, and relieve the repression effect caused by product accumulation in the straw carbon metabolism process.
Description
Technical Field
The invention belongs to the field of organic material decomposing agents or liquid bacterial manure, and particularly relates to a high-efficiency decomposing liquid bacterial manure and a preparation method and application thereof.
Background
As the related ministerial committees of the state issue policies such as straw burning prohibition and comprehensive resource utilization successively, the mode taking straw returning as priority is developed rapidly. The straws are agricultural byproducts with very high fiber component content, wherein the refractory macromolecular chemical components such as cellulose, hemicellulose and lignin have compact structures, complex structures, high polymerization degrees, stable properties and strong decomposition resistance, so that the decomposition rate of the returned straws under natural conditions is slow, and a large amount of straws which are not fully decomposed cause the problems of aggravated diseases and insect pests, influence on the growth of next-stubble crops and the like. The straw is biologically converted by utilizing related functional bacteria in nature, and the straw biological conversion agent has the characteristics of high degradation efficiency, safety, no pollution and the like.
At present, most of commercial microbial agents in the market are compounded by functional bacteria for degrading lignocellulose, and the problems of poor system stability, non-lasting effect and the like generally exist. When the straw is subjected to biotransformation by using microorganisms, because the carbon-nitrogen ratio of the straw is high, in the process of straw carbon transformation, the microorganisms need to absorb nitrogen to maintain the growth and metabolism of the straw, while the microbial inoculum is usually sprayed on the surface of the straw, the available nitrogen is limited to the release of the straw nitrogen, the utilization of the nitrogen in soil is relatively difficult, and the problem of weak colonization of functional degradation bacteria after the microbial inoculum is applied is caused; on the other hand, the functional degradation bacteria can be subjected to a repression effect caused by accumulation of metabolites in the straw degradation process, and due to lack of lignocellulose intermediate metabolite degradation bacteria, imbalance in the straw carbon conversion process is easily caused, so that the straw decomposition function is difficult to be continuously exerted.
Disclosure of Invention
The invention mainly aims at the problem that the components of the straw microbial inoculum lack nitrogen sources and lignocellulose intermediate metabolic bacteria, and provides a preparation method of the high-efficiency decomposed liquid bacterial fertilizer, on one hand, the bacterial fertilizer liquid already contains quick-acting and slow-release nitrogen sources required by microorganism growth and metabolism, and the decomposition efficiency of the straw can be effectively improved without additionally supplementing the nitrogen sources during application; on the other hand, various metabolic pathways of different strains and complementation on different degrading enzyme systems are fused, pediococcus pentosaceus can effectively degrade intermediate metabolites of straw lignocellulose, such as low molecular oligosaccharides, lactic acid, ethanol and the like, and the repression effect caused by accumulation of the metabolites is relieved.
In order to solve the technical problems, a first objective of the present invention is to provide a high-efficiency decomposing liquid bacterial fertilizer, which contains three high-efficiency decomposing bacteria capable of secreting cellulase and xylanase: stenotrophomonas sp.WS6-1, achromobacter sp.WS6-2 and Pediococcus pentosaceus YC2, and the effective viable count in the bacterial manure is more than or equal to 1 × 109cfu/mL;
the Stenotrophomonas (Stenotrophoromonas sp.WS6-1) is preserved in China Center for Type Culture Collection (CCTCC) in 2019, 12 months and 18 days, and the preservation address is as follows: china, wuhan university, the preservation number is: CCTCC No. M20191060;
the Achromobacter sp.WS6-2 and the strain are preserved in China Center for Type Culture Collection (CCTCC) in 2019, 12 months and 18 days, and the preservation addresses are as follows: china, wuhan university, the preservation number is: CCTCC No. M20191061;
the Pediococcus pentosaceus (Pediococcus pentosaceus YC 2) is preserved in the China Center for Type Culture Collection (CCTCC) in 2019, 12 months and 18 days, and the preservation address is as follows: china, wuhan university, the preservation number is: CCTCC No. M20191062. When the liquid bacterial manure is applied, no additional nitrogen source is needed to be supplemented
The efficient decomposition liquid bacterial manure provided by the invention is further set to contain eupenicillium crustaceum which is purchased from Beijing Beinana Chuanglian Biotechnology research institute and has the resource number of BNCC146720. The cold-resistant eupenicillium crustaceum is adopted, the metabolic pathway and complementation on different degrading enzyme systems can effectively relieve the repression effect caused by accumulation of metabolites, and the decomposition speed is high.
The efficient decomposition liquid bacterial fertilizer provided by the invention is further set to contain one or more of bacillus subtilis, streptomyces roseoflavus, trichoderma harzianum and myceliophthora thermophila.
The efficient decomposition liquid bacterial fertilizer provided by the invention is further set to be that bacillus subtilis, streptomyces roseoflavus, trichoderma harzianum and myceliophthora thermophila in the bacterial fertilizer agent are purchased from Beijing Beinanna Chuanglian biotechnology research institute, the resource number of the bacillus subtilis is BNCC188080, the resource number of the streptomyces roseoflavus is BNCC228869, the resource number of the trichoderma harzianum is BNCC336568, and the resource number of the myceliophthora thermophila is BNCC186098.
The second purpose of the invention is to provide a preparation method of the high-efficiency decomposed liquid bacterial fertilizer, which comprises the following steps:
step 1, strain activation
Taking a proper amount of pure culture from a slant seed-preserving culture medium according to aseptic operation, and inoculating stenotrophomonas and achromobacter to an LB solid plate; the inoculated flat plate is inverted and is placed in a culture box to be cultured at the temperature of 26 to 30 ℃, and when a single bacterial colony grows out of bacteria, fungi sporulation is stable and uniform in size;
step 2, seed culture
Under the aseptic condition, picking single colonies of the activated stenotrophomonas and achromobacter in the step 1 by using inoculating loops, and respectively inoculating the single colonies to an LB liquid culture medium; after inoculation of each strain, shaking and culturing the strain in a shaking table at 26-30 ℃ and 150r/min, wherein the culture time is determined according to the growth curve of each strain, and strain seed liquid in the middle and later logarithmic growth stages is obtained;
step 3, enlargement culture
Transferring the seed liquid obtained in the step (2) into a liquid culture medium of a corresponding type according to the inoculation amount of 10-20% (V/V), performing propagation expansion, adjusting the inoculation and culture time according to the growth rate of each strain of bacteria so as to obtain a bacterial liquid with basically consistent concentration at the same time, and performing centrifugal concentration on the bacterial liquid subjected to propagation expansion still according to aseptic operation;
step 4, mixed fermentation
And (3) preparing the thalli/spores collected by centrifugal concentration of the strains in the step (3) into heavy suspension with consistent concentration by using sterile water, mixing according to a ratio of 1.
The preparation method of the high-efficiency decay liquid bacterial fertilizer provided by the invention is further set as that in the step 1, the strain activation also comprises the steps of taking a proper amount of pure culture from a slant strain-preserving culture medium according to aseptic operation, and inoculating the bacillus subtilis to an LB solid plate; inoculating Trichoderma harzianum, penicillium crusum and myceliophthora thermophila to a PDA solid plate; inoculating Streptomyces roseoflavus to a Gao's No. 1 solid plate; and (3) inoculating lactic acid bacteria to the basic solid culture medium, inverting the inoculated plate, and horizontally placing the plate in a culture box for culture at the temperature of 26-30 ℃, until a single bacterial colony grows out of the bacteria, and spores produced by the fungi are stable and uniform in size.
The preparation method of the high-efficiency decomposed liquid bacterial fertilizer provided by the invention is further set as that in the step 2, under the aseptic condition, the seed culture also comprises the step of selecting a single bacterial colony from the bacillus subtilis activated in the step 1 by using an inoculating loop, and inoculating the single bacterial colony to an LB liquid culture medium; adding a small amount of sterile water into a trichoderma harzianum, eupenicillium crustosum and myceliophthora thermophila solid flat plate, gently scraping lawn by using an inoculating loop to wash off spores, filtering by using sterile cotton to remove hyphae, and inoculating to a PDA liquid culture medium; inoculating streptomyces roseoflavus to a Gao's No. 1 liquid culture medium containing glass beads; inoculating lactobacillus to the MRS liquid culture medium; after inoculation of each strain, shaking and culturing the strain at 26-30 ℃ and 150r/min in a shaking table, wherein the culture time is determined according to the growth curve of each strain, and strain seed liquid in the middle and later logarithmic growth stages is obtained.
The preparation method of the high-efficiency decomposition liquid bacterial fertilizer provided by the invention is further set as follows, an LB liquid culture medium: 10g of peptone, 10g of beef extract, 5g of sodium chloride, 1000mL of distilled water, pH6.5 to 7, and 20g of agar when preparing a solid culture medium;
PDA culture medium: 200g of peeled potato cooking juice 1000ml, sucrose 20g, monopotassium phosphate 3g, magnesium sulfate 1.5g and vitamin B110mg, wherein the pH value is natural; when preparing a solid culture medium, 20g of agar is added;
gao 1 medium: 20g of soluble starch, 1g of potassium nitrate, 0.5g of dipotassium hydrogen phosphate, 0.5g of magnesium sulfate, 0.5g of sodium chloride, 0.01g of ferrous sulfate and 1000mL of distilled water, wherein 20g of agar is added when a solid culture medium is prepared.
The preparation method of the high-efficiency decomposition liquid bacterial manure provided by the invention is further set as follows, wherein the liquid fermentation culture medium comprises the following components: 0.5% tryptone, 0.5% NaCl, 0.2% CaC03, 0.1% yeast powder, 0.3% urea, 1000mL of corn stalk extract, natural pH, 121 ℃, and sterilizing for 20min.
The third purpose of the invention is to provide the application of the liquid bacterial manure for efficient decomposition, the liquid bacterial manure is applied to the rapid and efficient decomposition of the corn straws, and a nitrogen source is not required to be additionally supplemented when the liquid bacterial manure is applied.
Has the beneficial effects.
1. The high-efficiency decomposition liquid bacterial fertilizer provided by the invention contains three high-efficiency decomposition bacteria capable of secreting cellulase and xylanase: stenotrophomonas sp.WS6-1, achromobacter sp.WS6-2 and Pediococcus pentosaceus YC2, which are all screened and preserved by the laboratory, have no antagonistic action among all strains, meet the assembly conditions and improve the degradation capability to the cellulose.
2. The efficient decomposition liquid bacterial fertilizer provided by the invention is prepared by compounding and fermenting microbial bacteria, the effective viable count is more than or equal to 1 x 109cfu/mL, the performance and function are stable, the decomposition speed of the corn straws is high, and the efficient decomposition liquid bacterial fertilizer has high market popularization and application values.
3. The invention provides a preparation method of a multi-strain-compounded liquid bacterial fertilizer agent for efficiently decomposing corn straws. The degradation capability of the corn straws is the best, the degradation rate of the corn straws is improved by 35.0 percent compared with that of the microorganism, and the degradation rate of the corn straws is improved by 66.3 percent and 76.8 percent compared with that of the Diele and the decomposition agent 150 respectively.
4. The invention provides a preparation method of a multi-strain combined maize straw high-efficiency decomposition liquid bacterial manure agent, which is characterized in that bacterial manure liquid is fused with various metabolic pathways of different strains and complementation on different degradation enzyme systems, pediococcus pentosaceus can effectively degrade intermediate metabolites of straw lignocellulose, such as low-molecular oligosaccharide, lactic acid, ethanol and the like, and the repression effect caused by accumulation of the metabolites is relieved.
Drawings
FIG. 1 is a graph of weight loss data for filter paper.
FIG. 2 is a graph of straw weight loss data.
FIG. 3 is a flow chart of the preparation and application of the high-efficiency decomposed liquid bacterial manure.
The specific implementation mode is as follows:
the present invention will be described in detail and specifically by the following examples, and it should be understood that the following examples are included to provide a better understanding of the present invention and are not intended to limit the scope of the present invention.
The efficient decomposition liquid bacterial fertilizer selects the following 8 effective active microorganisms: bacillus subtilis (A), (B) and (C)Bacillus subtilis) Streptomyces roseoflavus (see)Streptomyces roseoflavus) Trichoderma harzianum (a)Trichoderma harzianum Rifai) Penicillium notatum (A. B.) and (B. C.) A. BEupenicillium crustaceum) Myceliophthora thermophila (Myceliophthora thermophila) Pediococcus pentosaceus (A)Pediococcus pentosaceus YC2) The number of effective viable bacteria in the bacterial fertilizer agent is more than or equal to 1 multiplied by 10 9 cfu/mL。
Example 1
Preparation and functional verification of corn straw bacterium fertilizer
The method comprises the steps of combining two key functional strains in the decomposition process of the straw with stenotrophomonas, achromobacter and lactic acid bacteria obtained by laboratory self-screening, namely bacillus subtilis (biocontrol bacteria), streptomyces roseoflavus (biocontrol bacteria), trichoderma harzianum (biocontrol bacteria), penicillium corticalis (cold-resistant type) and myceliophthora thermophila to carry out a plate confrontation test, wherein results show that no antagonistic action exists among the strains, and the prerequisite condition of combination is met.
The 8 strains are subjected to strain activation, seed culture, enlarged culture and mixed fermentation to prepare the liquid bacterial fertilizer.
According to aseptic operation, using sterile water to prepare bacterial manure agent and stenotrophomonas, achromobacter, bacillus subtilis, streptomyces roseoflavus, trichoderma harzianum, penicillium crustosum and myceliophthora thermophila amplification culture solution into bacterial suspension/spore solutions with consistent concentrations, respectively taking 5mL of the bacterial suspension/spore solutions to inoculate in a triangular flask filled with 45mL of enzyme production culture medium, carrying out shaking culture on a shaking table at 30 ℃ and 150r/min, and taking fermentation liquor to determine beta-glucosidase activity, endoglucanase activity, exoglucanase activity, filter paper enzyme activity and xylanase activity at different time periods.
Definition of enzyme activity: under the specified conditions, the enzyme amount of 1mL of enzyme solution catalyzing substrate hydrolysis to generate 1 mug of reducing sugar per minute is one enzyme activity unit and is recorded as U/mL.
Enzyme production culture medium: 0.5% peptone, 0.5% corn stalk powder, 0.5% NaCl, 0.2% CaC03, 0.1% yeast powder, distilled water 1000mL, natural pH, 121 ℃ sterilization for 20min.
According to aseptic operation, using sterile water to prepare bacterial fertilizer agent and stenotrophomonas, achromobacter, bacillus subtilis, streptomyces roseoflavus, trichoderma harzianum, penicillium epidermidis and myceliophthora thermophila amplification culture solution into bacterial suspension/spore solutions with consistent concentrations, taking 5mL of each bacterial suspension/spore solution to be inoculated into a culture medium which takes straws and filter paper as substrates, carrying out shaking culture on a shaking table at 30 ℃ and 150r/min, sampling every 48h, adding a proper amount of (5 to 0 1mL) of mixed solution of sulfuric acid and sodium nitrate to remove thalli and insoluble calcium carbonate, sieving by a 80-mesh sieve, slowly washing the residual substrates on the sieve by distilled water, drying in an 80- ℃ oven to constant weight, weighing, calculating the weight loss rate, and enabling 3 groups to be parallel.
The measurement results are shown in table 1, fig. 1 and fig. 2.
TABLE 1 results of enzyme activity measurement
As shown in Table 1 and figures 1 and 2, under the same inoculation amount and culture conditions, after fermentation for 10 days, the enzyme activity of the bacterial manure agent is basically higher than that of a single bacterial strain in the combination; the filter paper weight loss rate of the bacterial manure agent is 28.0 percent, which is 10.6 percent higher than that of the single bacterial strain myceliophthora thermophila with the best degradation effect on the filter paper in the combination; the weight loss rate of the corn straws reaches 38.85 percent, the weight loss rate is improved by 2.25 to 4 times compared with that of a single bacterial strain in the combination, and the bacterial fertilizer prepared by combining multiple strains has the advantages of fusing multiple metabolic pathways of different bacterial strains and complementation on different degrading enzyme systems, realizing rapid degradation of the substrate, and showing that the degradation capability of the cellulosic substrate is enhanced after the bacterial strains are combined.
Example 2
Liquid fermentation degradation comparative tests are carried out on the liquid bacterial manure agent prepared by the invention and three commercially available organic material decomposition agents, in the tests, the substrate dosage, the microbial inoculum size, the culture conditions and the like are kept consistent, and 3 times of repetition are set.
Nutrient solution: 0.5% peptone, 0.5% NaCl, 0.2% 3 0.1% yeast powder, natural pH
The test method comprises the following steps: weighing 1g of substrate, adding the substrate into triangular flasks filled with 45mL of nutrient solution, sterilizing at 121 ℃ for 20min, diluting the prepared bacterial fertilizer agent and a commercially available organic material decomposition agent with sterile water, adjusting the diluted bacterial fertilizer agent and the commercially available organic material decomposition agent into suspensions with consistent concentrations, inoculating 5mL of the bacterial fertilizer agent and the commercially available organic material decomposition agent into the suspensions, performing shaking culture at 30 ℃ and 150r/min for 10d, sampling, adding a proper amount of sulfuric acid-sodium nitrate mixed solution into the samples, washing to remove attached bacteria and calcium carbonate, cleaning with distilled water, filtering, and drying in an oven at 70 ℃ to constant weight. The weight loss of the straw was measured by the mass difference method, and the results are shown in Table 2.
TABLE 2 substrate weight loss ratio
Through test comparison, the degradation capability of the bacterial fertilizer agent prepared by the invention on four cellulosic substrates is higher than that of a commercial microbial inoculum Dierle and a decomposition agent 150, the degradation capability on corn straws is the best among different cellulosic substrates, the degradation rate of the corn straws is improved by 35.0% compared with that of a microorganism, and the degradation rate of the corn straws is respectively improved by 66.3% and 76.8% compared with that of the Dierle and the decomposition agent 150.
Example 3
The liquid bacterial manure is applied to the rapid and efficient decomposition of rice, wheat and corn straws, the preferable field application temperature range is 5-25 ℃, and in addition, a nitrogen source does not need to be additionally supplemented when the liquid bacterial manure is applied. The application method of the liquid bacterial manure is to spray the liquid bacterial manure on the surface of the straw and perform heap fermentation. The fermentation method adopts conventional fermentation means and devices, and has the advantages of simple operation, convenient use and easy popularization and use.
The above is only a preferred embodiment of the present invention, and the protection scope of the present invention is not limited to the examples, and all technical solutions belonging to the idea of the present invention belong to the protection scope of the present invention. It should be noted that modifications and embellishments within the scope of the invention may occur to those skilled in the art without departing from the principle of the invention, and are considered to be within the scope of the invention.
Claims (9)
1. The efficient decomposition liquid bacterial fertilizer is characterized by comprising three efficient decomposition bacteria capable of secreting cellulase and xylanase: stenotrophomonas sp.WS6-1, achromobacter sp.WS6-2, pediococcus pentosaceus YC2, and Penicillium corticalis, bacillus subtilis, streptomyces roseoflavus, trichoderma harzianum, and myceliophthora thermophila, wherein the Stenotrophomonas sp.WS6-1, achromobacter sp.WS6-2, pediococcus pentosaceus YC2, penicillium corticalis, bacillus subtilis, streptomyces roseoflavus, trichoderma harzianum, and myceliophthora thermophila are in the same weight parts, and the number of effective viable bacteria in the bacterial manure is not less than 1 × 10 9 cfu/mL；
The Stenotrophomonas (Stenotrophoromonas sp.WS6-1) is preserved in China Center for Type Culture Collection (CCTCC) in 2019, 12 and 18 months, and the preservation number is as follows: CCTCC No.20191060;
the Achromobacter (Achromobacter sp.WS6-2) is preserved in China Center for Type Culture Collection (CCTCC) in 2019, 12 and 18 months, and the preservation number is as follows: CCTCC No.20191061;
the Pediococcus pentosaceus (Pediococcus pentosaceus YC 2) is preserved in the China Center for Type Culture Collection (CCTCC) in 2019, 12 months and 18 days, and the preservation number is as follows: CCTCC No.20191062..
2. The efficient decomposition liquid bacterial fertilizer according to claim 1, wherein the Penicillium corticium is purchased from the institute of biotechnology, chuanglian and Biotechnology, beijing, and has a resource number of BNCC146720.
3. The efficient liquid bacterial fertilizer for decay according to claim 1, wherein the bacillus subtilis, the streptomyces roseoflavus, the trichoderma harzianum and the myceliophthora thermophila in the bacterial fertilizer are all purchased from Beijing BeiNayiniaoSou Biotech research institute, and the resource number of the bacillus subtilis is BNCC188080, the resource number of the streptomyces roseoflavus is BNCC228869, the resource number of the trichoderma harzianum is BNCC336568, and the resource number of the myceliophthora thermophila is BNCC186098.
4. A method for preparing the high-efficiency decomposed liquid bacterial fertilizer as claimed in any one of claims 1 to 3, which is characterized by comprising the following steps:
step 1, strain activation
Taking a proper amount of pure culture from a slant culture preserving medium according to aseptic operation, and inoculating stenotrophomonas and achromobacter to an LB solid plate; the inoculated flat plate is inverted and is placed in a culture box to be cultured at the temperature of 26 to 30 ℃, and when a single bacterial colony grows out of bacteria, fungi sporulation is stable and uniform in size;
step 2, seed culture
Under the aseptic condition, picking single colonies of the activated stenotrophomonas and achromobacter in the step 1 by using inoculating loops, and respectively inoculating the single colonies to an LB liquid culture medium; after inoculation of each strain, shaking and culturing the strain in a shaking table at 26-30 ℃ and 150r/min, wherein the culture time is determined according to the growth curve of each strain, and strain seed liquid in the middle and later logarithmic growth stages is obtained;
step 3, enlargement culture
Transferring the seed solution obtained in the step (2) into a liquid culture medium of a corresponding type according to the inoculation amount of 10-20% (V/V), performing propagation expansion, adjusting the inoculation time and the culture time according to the growth rate of each strain of bacteria so as to obtain a bacterial solution with consistent concentration at the same time, and performing centrifugal concentration on the bacterial solution subjected to propagation expansion still according to aseptic operation;
step 4, mixed fermentation
And (3) centrifugally concentrating and collecting the strains/spores obtained in the step (3), preparing the strains/spores into heavy suspension with consistent concentration by using sterile water, mixing the strains/spores according to a ratio of 1 9 cfu/mL liquid bacterial manure.
5. The method for preparing the high-efficiency decomposed liquid bacterial fertilizer as claimed in claim 4, wherein the method comprises the following steps: step 1, the strain activation also comprises taking a proper amount of pure culture from a slant strain preservation culture medium according to aseptic operation, and inoculating the bacillus subtilis to an LB solid plate; inoculating trichoderma harzianum, eupenicillium crustosum and myceliophthora thermophila to a PDA solid plate; inoculating Streptomyces roseoflavus to a Gao's No. 1 solid plate; inoculating pediococcus pentosaceus to a basic solid culture medium, inverting the inoculated plate, and putting the plate in a culture box for culture at the temperature of 26-30 ℃, until a single bacterial colony grows out of the bacteria, and spores produced by the fungi are stable and uniform in size.
6. The method for preparing the efficient decomposed liquid bacterial fertilizer as claimed in claim 5, wherein the seed culture in step 2 further comprises the steps of picking up a single colony of the bacillus subtilis activated in step 1 by using an inoculating loop under aseptic conditions, and inoculating the single colony to an LB liquid culture medium; adding a small amount of sterile water into a trichoderma harzianum, eupenicillium crustosum and myceliophthora thermophila solid flat plate, gently scraping lawn by using an inoculating loop to wash off spores, filtering by using sterile cotton to remove hyphae, and inoculating to a PDA liquid culture medium; inoculating streptomyces roseoflavus to a Gao's No. 1 liquid culture medium containing glass beads; inoculating pediococcus pentosaceus to an MRS liquid culture medium; after inoculation of each strain, shaking and culturing the strain in a shaking table at the temperature of 26-30 ℃ and at the speed of 150r/min, wherein the culture time is determined according to the growth curve of each strain, and thus strain seed liquid in the middle and later logarithmic growth stages is obtained.
7. The method for preparing high efficiency liquid bacterial manure decay as claimed in claim 6, wherein,
LB liquid medium: 10g of peptone, 10g of beef extract, 5g of sodium chloride, 1000mL of distilled water, 6.5-7 of pH, and 20g of agar when preparing a solid culture medium;
PDA culture medium: 200g of peeled potato cooking juice 1000ml, sucrose 20g, monopotassium phosphate 3g, magnesium sulfate 1.5g and vitamin B110mg, and the pH is natural; when preparing a solid culture medium, 20g of agar is added;
gao 1 medium: 20g of soluble starch, 1g of potassium nitrate, 0.5g of dipotassium hydrogen phosphate, 0.5g of magnesium sulfate, 0.5g of sodium chloride, 0.01g of ferrous sulfate and 1000mL of distilled water, wherein 20g of agar is added when a solid culture medium is prepared.
8. The method for preparing the efficient decomposed liquid bacterial fertilizer according to claim 6, wherein the liquid fermentation culture medium comprises: 0.5% tryptone, 0.5% NaCl, 0.2% CaC03, 0.1% yeast powder, 0.3% urea, 1000mL of corn stalk extract, natural pH, 121 ℃, and sterilizing for 20min.
9. The application of the high-efficiency decomposed liquid bacterial manure as claimed in any one of claims 1-3, wherein the liquid bacterial manure is applied to the rapid high-efficiency decomposition of corn stalks, and a nitrogen source is not required to be additionally supplemented when the liquid bacterial manure is applied.
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