CN115369044B - Soil decomposing inoculant for degrading straw and application thereof - Google Patents

Soil decomposing inoculant for degrading straw and application thereof Download PDF

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CN115369044B
CN115369044B CN202210839134.7A CN202210839134A CN115369044B CN 115369044 B CN115369044 B CN 115369044B CN 202210839134 A CN202210839134 A CN 202210839134A CN 115369044 B CN115369044 B CN 115369044B
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straw
decomposing inoculant
culture medium
inoculant
liquid
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CN115369044A (en
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张德健
李冠华
安妮
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Inner Mongolia University
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    • 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
    • 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
    • C—CHEMISTRY; METALLURGY
    • C05—FERTILISERS; MANUFACTURE THEREOF
    • C05F—ORGANIC FERTILISERS NOT COVERED BY SUBCLASSES C05B, C05C, e.g. FERTILISERS FROM WASTE OR REFUSE
    • C05F17/00—Preparation of fertilisers characterised by biological or biochemical treatment steps, e.g. composting or fermentation
    • C05F17/20—Preparation of fertilisers characterised by biological or biochemical treatment steps, e.g. composting or fermentation using specific microorganisms or substances, e.g. enzymes, for activating or stimulating the treatment
    • C—CHEMISTRY; METALLURGY
    • C05—FERTILISERS; MANUFACTURE THEREOF
    • C05F—ORGANIC FERTILISERS NOT COVERED BY SUBCLASSES C05B, C05C, e.g. FERTILISERS FROM WASTE OR REFUSE
    • C05F17/00—Preparation of fertilisers characterised by biological or biochemical treatment steps, e.g. composting or fermentation
    • C05F17/50—Treatments combining two or more different biological or biochemical treatments, e.g. anaerobic and aerobic treatment or vermicomposting and aerobic treatment
    • 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
    • C05G3/80—Soil conditioners
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    • 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
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    • C12R2001/00—Microorganisms ; Processes using microorganisms
    • C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
    • C12R2001/07—Bacillus
    • C12R2001/11—Bacillus megaterium
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    • C12R2001/00—Microorganisms ; Processes using microorganisms
    • C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
    • C12R2001/07—Bacillus
    • C12R2001/125—Bacillus subtilis ; Hay bacillus; Grass bacillus
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    • C12R2001/00—Microorganisms ; Processes using microorganisms
    • C12R2001/645—Fungi ; Processes using fungi
    • 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

The invention discloses a soil decomposing inoculant for degrading straws and application thereof. The active ingredients of the soil decomposing inoculant for degrading straw comprise: the preparation method comprises the steps of preparing a Inonotus obliquus decomposing inoculant, a birch shell tube fungus decomposing inoculant, a bacillus subtilis decomposing inoculant and a phosphate-solubilizing bacillus megaterium decomposing inoculant. The soil decomposing inoculant can be applied to field decomposition of corn stalks, straw stalks, wheat stalks and sorghum stalks, and the stalk degradation rate can reach 60-80%.

Description

Soil decomposing inoculant for degrading straw and application thereof
Technical Field
The invention relates to the technical field of agricultural microorganisms, in particular to a soil decomposing inoculant for degrading straws and application thereof.
Background
The effective utilization of biomass resources is an important measure for relieving energy crisis, reducing greenhouse gas emission and promoting social sustainable development. Straw is the residual product of crops after harvest and is also an important organic resource for farmland ecosystems. The agricultural crop straw is rich in various crop straw resources, and along with development of rural economy and solution of rural energy problems, the straw is no longer used as a main fuel, and the phenomenon of concentrated burning is increasingly prominent in harvesting seasons, so that adverse effects are brought to the quality of ambient air. In recent years, how to utilize straw has become a research hot spot, wherein straw returning is the first choice, because straw returning can return nutrients to soil, increase soil organic matters and improve soil quality.
In the main components of the straw, the cellulose content accounts for 30% -35%, the hemicellulose content accounts for 25% -30%, the lignin content accounts for 20% -25%, and the components are degraded by external microorganisms. The microorganism has the characteristics of simple production flow, strong adaptability and the like, becomes an important participant for straw degradation, and is increasingly focused in agriculture at present, especially straw decomposition due to the important role in straw decomposition.
The straw decomposing inoculant is an agricultural microbial inoculant developed by the method, and the effective bacteria are viable bacteria preparations prepared by industrial production and propagation. The agricultural microbial inoculant comprises rhizobium inoculant, azotobacter inoculant, phosphate-solubilizing microbial inoculant, silicate microbial inoculant, photosynthetic inoculant, organic material decomposition agent, growth promoting inoculant, mycorrhizal inoculant, bioremediation inoculant and the like. The straw decomposing inoculant is one of organic material decomposing inoculants. The straw decomposing inoculant contains a large amount of thermophilic, heat-resistant and actinomycetes and biological enzymes which can strongly decompose cellulose, hemicellulose and lignin, and the mass propagation of the straw decomposing inoculant can effectively decompose crop straw into small organic components or inorganic components and return the crop straw to soil, so that the straw decomposing inoculant is favorable for improving the organic content of the soil, and simultaneously, releases a large amount of elements such as nitrogen, phosphorus, potassium and the like and medium and trace elements such as calcium, magnesium, manganese, molybdenum and the like required by the growth of crops. The decomposed organic matters return to the soil, so that the soil aggregate structure can be effectively improved, the soil ventilation, fertilizer and water retention functions can be improved, heat and a certain amount of carbon dioxide can be generated, the growth environment of plants can be improved, and the growth of crops can be promoted.
However, due to the complex, tough and high heterogeneity of straw components, the degradation enzyme systems required by different crop straws in the straw decomposition process may be different, and the microbial community has obvious succession phenomena in different periods of straw degradation. Therefore, the existing straw decomposing inoculant has the defects in the aspects of applicability and stage aiming at different crop straws, and influences the degradation effect of the agricultural straws.
Disclosure of Invention
Based on the problems of low efficiency and long degradation period of the existing straw-decomposing inoculant for agriculture as straw, the soil-decomposing inoculant which has universality for different crop straws and can meet the requirements of different degradation stages is provided.
The soil decomposing inoculant for degrading straw is a composite inoculant, and the active ingredients of the soil decomposing inoculant comprise: the preparation method comprises the steps of preparing a Inonotus obliquus decomposing inoculant, a birch shell tube fungus decomposing inoculant, a bacillus subtilis decomposing inoculant and a phosphate-solubilizing bacillus megaterium decomposing inoculant.
In one embodiment, the suspension of the decomposing inoculant spores of the phellinus linteus is 10 5 ～10 7 The spore suspension content of the decomposed fungus agent of the inonotus obliquus per ml is 10 5 ～10 7 The cfu content of the bacillus subtilis decomposed microbial agent per ml is 10 5 ～10 8 The cfu content of the decomposed microbial agent of the bacillus megaterium per ml is 10 5 ～10 8 And each ml.
In one embodiment, the sclerotium rolfsii decomposing inoculant is prepared by the following method:
(1) Seed culture medium: glucose 1%, malt extract 1%, initial pH 5;
(2) Liquid medium: glucose 0.5%, starch 1%, peptone 1%, copper sulfate 0.005% and initial pH 5;
(3) Seed preparation: 3 bacterial sheets with the length of 5-7 mm are inoculated in a seed culture medium, and liquid fermentation is carried out for 12-48 hours at the temperature of 25-32 ℃;
(4) Alkali-treating the straw carrier matrix, adding a liquid culture medium, and sterilizing at 121 ℃ for 20min to obtain a solid culture medium;
(5) Inoculating seed liquid into the solid culture medium, controlling the water content to be 55-80% by 0.5ml of seed/gram of straw carrier matrix, and performing aerobic fermentation for 5-7 days at the temperature of 25-32 ℃ to obtain the sclerotium rolfsii decomposing inoculant.
In one embodiment, the birch-leaf tube hole fungus decomposing inoculant is prepared by the following method:
(1) Seed culture medium: glucose 1%, malt extract 1%, initial pH 5;
(2) Liquid medium: glucose 1%, peptone 1%, initial pH 5;
(3) Seed preparation: 3 bacterial sheets with the length of 5-7 mm are inoculated in a seed culture medium, and liquid fermentation is carried out for 12-48 hours at the temperature of 25-32 ℃;
(4) Alkali-treating the straw carrier matrix, adding a liquid culture medium, and sterilizing at 121 ℃ for 20min to obtain a solid culture medium;
(5) Inoculating seed liquid into the solid culture medium, controlling the water content to be 55-80% by 0.5ml of seed/gram of straw carrier matrix, and performing aerobic fermentation for 5-7 days at 25-32 ℃ to obtain the birch tube-peeling fungus decomposing inoculant.
In one embodiment, the bacillus subtilis decomposing inoculant is prepared by the following method:
(1) Seed culture medium: 1% of peptone, 0.5% of yeast extract, 1% of NaCL and natural pH;
(2) Liquid medium: glucose 0.5%, peptone 1%, yeast extract 0.5%, naCL1% and natural pH;
(3) Seed preparation: b, selecting a bacillus subtilis colony 3 ring, inoculating the bacillus subtilis colony to seed liquid, and carrying out liquid fermentation for 6-36 h by a shaking table at 37 ℃ and 150 rpm;
(4) Alkali-treating the straw carrier matrix, adding a liquid culture medium, and sterilizing at 121 ℃ for 20min to obtain a solid culture medium;
(5) Inoculating seed liquid into the solid culture medium, controlling the water content to be 55-80% by the inoculum size of 0.5ml seed/gram straw carrier matrix, and carrying out aerobic fermentation for 24-48 h at 37 ℃ to obtain the bacillus subtilis decomposed microbial inoculum.
In one embodiment, the bacillus megaterium decomposing inoculant is prepared by the following steps:
(1) Seed culture medium: 1% of peptone, 0.5% of yeast extract, 1% of NaCL and natural pH;
(2) Liquid medium: glucose 0.5%, peptone 1%, yeast extract 0.5%, naCL1% and natural pH;
(3) Seed preparation: b, selecting a bacillus subtilis colony 3 ring, inoculating the bacillus subtilis colony to seed liquid, and carrying out liquid fermentation for 6-36 h by a shaking table at 37 ℃ and 150 rpm;
(4) Alkali-treating the straw carrier matrix, adding a liquid culture medium, and sterilizing at 121 ℃ for 20min to obtain a solid culture medium;
(5) Inoculating seed liquid into the solid culture medium, controlling the water content to be 55-80% by the inoculum size of 0.5ml seed/gram straw carrier matrix, and carrying out aerobic fermentation for 24-48 h at 37 ℃ to obtain the phosphate-dissolving bacillus megatherium decomposed microbial agent.
In one embodiment, the alkali treated straw carrier matrix is prepared by the following method: crushing straw, sieving with a 20-60 mesh sieve, adding 0.5% -1.5% NaOH solution, reacting at 50-80 ℃ for 0.5-1.5 h at a solid-liquid ratio of 1:4-1:6, and flushing with distilled water to pH 6 after the reaction is finished.
The invention also provides application of the soil decomposing inoculant for degrading the straws in field decomposition of the farmland straws.
In one embodiment, when the soil-decomposing inoculant is applied to field decomposition of farmland straws, sequentially inoculating the soil-decomposing inoculant, the birch-leaf-tube hole fungus-decomposing inoculant, the bacillus subtilis-decomposing inoculant and the phosphorus-decomposing bacillus megaterium-decomposing inoculant in the order of 0.5-1 g inoculant per 100g straw.
In one embodiment, the soil decomposing inoculant is applied to field straw decomposition in the following specific modes:
(1) Inoculating a sclerotium rolfsii decomposing inoculant, decomposing for 5-7 days, and rolling for 1-3 times/day;
(2) Inoculating a decomposed inoculant of the birch tube-peeling fungus, decomposing for 5-7 days, and rolling for 1-3 times/day;
(3) Inoculating bacillus subtilis decomposing inoculant, decomposing for 2-3 days, and rolling for 1-3 times/day;
(4) Inoculating Bacillus megaterium decomposing inoculant, and simultaneously supplementing 1g/100g of urea and decomposing for 20 days.
The straw refers to corn straw, straw stalk, wheat straw stalk and sorghum straw stalk.
The basic conditions of the microorganism strain according to the present invention are as follows:
in the invention, the adopted Inonotus obliquus (Funalia trogii), inonotus obliquus (Piptoporus betulinus), bacillus subtilis (Bacillus subtilis) and bacillus megaterium (Bacillus megaterium) are all purchased from China general microbiological culture Collection center.
Tuber crudus (Funalia trogii): the white rot fungi which are grown on broad-leaved tree branches or stumps of poplar, willow and the like are common species in the north, belonging to basidiomycete, mushroom class, polyporaceae, pileus, and the like, can efficiently decompose lignin and simultaneously degrade cellulose and hemicellulose.
Inonotus obliquus (Piptoporus betulinus): also known as Inonotus obliquus and Inonotus obliquus, belonging to the genus Basidiomycota, agaricales, polyporales, and Pelargonium, the species commonly used in northeast China, the fruit body forms in summer and autumn, and is frequently generated on living birch and inverted wood, cellulose and hemicellulose are mainly decomposed, and lignin is modified, so that brown rot caused by wood can be rapidly degraded.
Bacillus subtilis (Bacillus subtilis): belonging to the genus Bacillus, the order Bacillus, the family Bacillus and the genus Bacillus, belonging to the phylum Thick-walled bacteria, is a common soil bacterium, widely distributed in soil and putrefactive organic matters, and has strong resistance.
Bacillus megaterium phosphate (Bacillus megaterium): belonging to the genus Bacillus, the order Bacillus, the family Bacillus and the genus Bacillus, belonging to the phylum Thick-walled bacteria, is a common soil bacterium, can decompose soil phosphide, and effectively solves the problem of soil salinization.
The invention starts from the intrinsic structure of the chemical composition of the straw, simulates the mechanism of natural decomposition of the straw by microorganisms, adopts the concept of continuous and sequential decomposition, and orderly compounds four microorganisms according to the physical and chemical properties of the straw to construct the straw decomposing inoculant. Wherein, the bristle covered porus can secrete laccase, oxidase such as peroxidase, etc., and oxidize lignin to form micromolecular compounds; the inonotus obliquus can secrete hydrolytic enzymes such as cellulase, hemicellulase and the like, can decompose cellulose and hemicellulose, oxidize lignin and improve the hydroxyl content of lignin; the bacillus subtilis has the effects of growing rapidly and regulating soil microbial flora, and can improve the soil microbial flora composition while rapidly utilizing the organic matters; the bacillus megaterium has the function of phosphate dissolution, has obvious decomposition effect on organic phosphorus and inorganic phosphorus which cannot be directly absorbed and utilized by plants in lecithin and soil, can convert the phosphorus which is difficult to be absorbed and utilized by the plants into a form which can be absorbed and utilized, and can improve the soil fertility. The four microorganisms have synergistic effect in the composite microbial agent, so that the effect of quick decomposition of straw can be realized, and the soil fertility are improved.
The four methods are used for respectively preparing the sclerotium rolfsii decomposing inoculant, the inonotus obliquus decomposing inoculant, the bacillus subtilis decomposing inoculant and the bacillus megaterium decomposing inoculant, so that the microbial nutrition requirement is low, the culture medium is cheap, common and easy to obtain, the environmental pollution is avoided, the microbial growth speed is high, and the culture condition is simple.
The carrier matrix adopted by the invention has the advantages of wide sources of raw materials, low price and reutilization of wastes in the process of preparing the carrier matrix by conversion; on the other hand, the straw serving as a carrier has strong biodegradability, is farmland crops per se, can be rapidly decomposed under natural conditions, and cannot produce secondary pollution to farmland soil. The preparation method adopts the alkaline pretreatment to prepare the carrier matrix, is simple and convenient to operate and has low production cost; the specific surface area of the obtained carrier matrix is remarkably improved, more microbial agents can be attached, the interaction between the microorganisms and the carrier matrix is tighter, and the microorganisms can rapidly provide nutrition for the microbial agents after entering the soil, so that the microorganisms rapidly grow, and the influence of severe environment on the microorganisms is relieved; in addition, compared with the liquid microbial inoculum, the solid carrier has better preservation effect, is easier to transport and easier to grow microorganisms, and accords with the current novel concepts of green and clean development.
The invention sequentially uses the seed-metering sequence of the Inonotus obliquus decomposing inoculant, the Bacillus subtilis decomposing inoculant and the Bacillus megaterium decomposing inoculant, and the Inonotus obliquus decomposing inoculant can quickly utilize cellulose and hemicellulose as nutrients to decompose the substances to generate monosaccharide, and laccase, manganese peroxidase and the like generated in the process can oxidize and break lignin to decompose the lignin into micromolecular compounds, and simultaneously expose a cellulose crystallization area. The inonotus obliquus can secrete cellulase and the like, the enzyme can further decompose an exposed cellulose crystallization area, and simultaneously, the lignin micromolecular substance is subjected to oxidation modification, so that lignin is further hydroxylated, and the water solubility is improved. The micromolecular saccharide compound produced by decomposing the two bacteria can be further used as a nutrient substance for the growth and utilization of bacillus subtilis and phosphate-dissolving bacillus megaterium, the two bacillus can grow rapidly and consume the substances, a small amount of micromolecular inhibitor is produced in the growth process of the bacillus subtilis, the growth of harmful microorganisms such as soil parasites and escherichia coli is further inhibited, on the other hand, the phosphate-dissolving bacillus can decompose inorganic phosphate radicals, the hydrolysis of inorganic phosphorus in the soil is promoted, and the dissolution of calcium ions is promoted, so that the phosphate-dissolving bacillus megaterium can be used for absorbing organic phosphorus and calcium.
By applying the soil decomposing inoculant disclosed by the invention to field straw decomposition, the straw degradation rate can reach 60% -80%.
Compared with the prior art, the invention has the following advantages:
(1) Aiming at the intrinsic characteristics of straw, the soil decomposing inoculant provided by the invention realizes rapid decomposition of straw components in different compounding modes, different components are utilized in a targeted manner, cellulose and hemicellulose are mainly decomposed into small molecular sugar to provide nutrition for other microorganisms, and other microorganisms grow by utilizing the small molecular sugar, so that the effects of improving the composition of soil microorganisms and improving soil nutrients are realized.
(2) In the process of straw degradation by using the soil decomposing inoculant, lignin which is a product of straw decomposition is changed into a micromolecular compound, so that the water solubility is improved, and the lignin can be used as soil humic substances to further improve the soil fertility.
(3) The carrier matrix used in the invention is derived from crop residues, has wide sources and low price, does not generate new pollution, has better carrier adsorption performance, can enable microorganisms to grow on the carrier more easily, and has the advantages of easy storage, easy transportation and the like.
Detailed Description
The following detailed description of the present invention will provide further details in order to make the above-mentioned objects, features and advantages of the present invention more comprehensible. In the following description, numerous specific details are set forth in order to provide a thorough understanding of the present invention. The present invention may be embodied in many other forms than described herein and similarly modified by those skilled in the art without departing from the spirit of the invention, whereby the invention is not limited to the specific embodiments disclosed below.
Example 1 preparation of soil-decomposing inoculant
1-1 preparation of a straw carrier matrix by alkali treatment: crushing straw, sieving with a 40-mesh sieve, adding 1% NaOH solution, reacting for 1h at the solid-liquid ratio of 1:5 and 65 ℃, and flushing with distilled water to pH 6 after the reaction is finished.
1-2 preparation of a thick california cerealis decomposing inoculant:
(1) Seed culture medium: glucose 1%, malt extract 1%, initial pH 5;
(2) Liquid medium: glucose 0.5%, starch 1%, peptone 1%, copper sulfate 0.005% and initial pH 5;
(3) Seed preparation: 36 mm fungus slices are inoculated in a seed culture medium, and liquid state fermentation is carried out for 30 hours at the temperature of 28 ℃;
(4) Alkali-treating the straw carrier matrix, adding a liquid culture medium, and sterilizing at 121 ℃ for 20min to obtain a solid culture medium;
(5) Inoculating seed solution into the solid culture medium, controlling the water content at 70% and 28 ℃ with the inoculum size of 0.5ml seed/gram straw carrier matrix, and performing aerobic fermentation for 6 days to obtain the sclerotium rolfsii decomposing inoculant.
1-3 preparation of a birch tube-peeling fungus decomposing inoculant:
(1) Seed culture medium: glucose 1%, malt extract 1%, initial pH 5;
(2) Liquid medium: glucose 1%, peptone 1%, initial pH 5;
(3) Seed preparation: 36 mm fungus slices are inoculated in a seed culture medium, and liquid state fermentation is carried out for 30 hours at the temperature of 28 ℃;
(4) Alkali-treating the straw carrier matrix, adding a liquid culture medium, and sterilizing at 121 ℃ for 20min to obtain a solid culture medium;
(5) Inoculating seed solution into the solid culture medium, controlling the water content at 70% and 28 ℃ with the inoculum size of 0.5ml seed/gram straw carrier matrix, and performing aerobic fermentation for 6 days to obtain the birch shell tube fungus decomposing inoculant.
1-4 preparation of bacillus grass decomposing inoculant:
(1) Seed culture medium: 1% of peptone, 0.5% of yeast extract, 1% of NaCL and natural pH;
(2) Liquid medium: glucose 0.5%, peptone 1%, yeast extract 0.5%, naCL1% and natural pH;
(3) Seed preparation: b, picking a bacillus subtilis colony 3 ring, inoculating the bacillus subtilis colony to a seed solution, and carrying out liquid fermentation for 21h by a shaking table at 37 ℃ and 150 rpm;
(4) Alkali-treating the straw carrier matrix, adding a liquid culture medium, and sterilizing at 121 ℃ for 20min to obtain a solid culture medium;
(5) Inoculating seed liquid into the solid culture medium, controlling the water content at 70% and the temperature at 37 ℃ in an inoculum size of 0.5ml seed/gram straw carrier matrix, and performing aerobic fermentation for 36 hours to obtain the bacillus subtilis decomposed microbial inoculum.
1-5 preparation of a bacillus megatherium decomposing inoculant:
(1) Seed culture medium: 1% of peptone, 0.5% of yeast extract, 1% of NaCL and natural pH;
(2) Liquid medium: glucose 0.5%, peptone 1%, yeast extract 0.5%, naCL1% and natural pH;
(3) Seed preparation: b, picking a bacillus subtilis colony 3 ring, inoculating the bacillus subtilis colony to a seed solution, and carrying out liquid fermentation for 21h by a shaking table at 37 ℃ and 150 rpm;
(4) Alkali-treating the straw carrier matrix, adding a liquid culture medium, and sterilizing at 121 ℃ for 20min to obtain a solid culture medium;
(5) Inoculating seed liquid into the solid culture medium, controlling the water content at 70% and the temperature at 37 ℃ in an inoculum size of 0.5ml seed/gram straw carrier matrix, and performing aerobic fermentation for 36 hours to obtain the bacillus megaterium decomposing inoculant.
Example 2 field straw decomposition test
Pulverizing the aerial parts of the harvested corn stalks, controlling the water content to 55-80%, and sequentially inoculating the sclerotium rolfsii decomposing inoculant (the spore suspension content is 4.5X10) 5 The content of spore suspension is 6.5X10 6 Each ml), bacillus subtilis decomposed microbial agent (cfu content is 2.8X10) 7 Bacillus megaterium decomposing inoculant (cfu content is 4.9X10) 6 Per ml), the inoculum size is 0.75g microbial inoculum per 100g straw.
The specific operation steps are as follows:
(1) Inoculating a sclerotium rolfsii decomposing inoculant, decomposing for 5-7 days, and rolling for 1-3 times/day;
(2) Inoculating a decomposed inoculant of the birch tube-peeling fungus, decomposing for 5-7 days, and rolling for 1-3 times/day;
(3) Inoculating bacillus subtilis decomposing inoculant, decomposing for 2-3 days, and rolling for 1-3 times/day;
(4) Inoculating Bacillus megaterium decomposing inoculant, and simultaneously supplementing 1g/100g of urea and decomposing for 20 days.
Collecting decomposed straw, washing with distilled water, removing microbial mycelium on the surface of the substrate as much as possible, drying in a 63 ℃ oven to constant weight, and calculating the mass loss rate in the straw decomposition process.
The mass loss rate calculation formula is as follows:
y represents the mass loss rate in straw decomposition, w i Represents the initial mass of straw, w f Representing the quality of the straw after decomposition.
The calculation formula of the loss rate of a specific component in the straw before and after decomposition is as follows:
wherein Y is s Represents the mass loss rate of a certain component in the straw, X i Representing the initial content of a certain component in the straw, and Xf represents the decomposed content of the certain component in the straw.
The degradation rate of the corn straw is 77 percent by the method.
The technical features of the above-described embodiments may be arbitrarily combined, and all possible combinations of the technical features in the above-described embodiments are not described for brevity of description, however, as long as there is no contradiction between the combinations of the technical features, they should be considered as the scope of the description.
The above examples illustrate only a few embodiments of the invention, which are described in detail and are not to be construed as limiting the scope of the invention. It should be noted that it will be apparent to those skilled in the art that several variations and modifications can be made without departing from the spirit of the invention, which are all within the scope of the invention. Accordingly, the scope of protection of the present invention is to be determined by the appended claims.

Claims (9)

1. The soil-decomposing inoculant for degrading straw is characterized by comprising the following active ingredients: the spore suspension of the Inonotus obliquus decomposing inoculant is 10 5 ~10 7 The spore suspension content of the decomposed fungus agent of the inonotus obliquus per ml is 10 5 ~10 7 The cfu content of the bacillus subtilis decomposed microbial agent per ml is 10 5 ~10 8 The cfu content of the decomposed microbial agent of the bacillus megaterium per ml is 10 5 ~10 8 And each ml.
2. The straw degrading soil-decomposing inoculant of claim 1, wherein the straw degrading inoculant is prepared by the following steps:
(1) Seed culture medium: glucose 1%, malt extract 1%, initial pH 5;
(2) Liquid medium: glucose 0.5%, starch 1%, peptone 1%, copper sulfate 0.005% and initial pH 5;
(3) Seed preparation: 3 bacterial slices with the length of 5-7 mm are inoculated in a seed culture medium, and liquid fermentation is carried out for 12-48 hours at the temperature of 25-32 ℃;
(4) Alkali-treating the straw carrier matrix, adding a liquid culture medium, and sterilizing at 121 ℃ for 20min to obtain a solid culture medium;
(5) Inoculating seed liquid into the solid culture medium, controlling the water content to be 55% -80% by inoculating 0.5ml of seeds per gram of straw carrier matrix, and performing aerobic fermentation at 25 ℃ -32 ℃ for 5-7 days to obtain the rotten inoculant for the porus rupestris.
3. The straw-degrading soil-decomposing inoculant of claim 1, wherein the birch-leaf fungus-decomposing inoculant is prepared by the following steps:
(1) Seed culture medium: glucose 1%, malt extract 1%, initial pH 5;
(2) Liquid medium: glucose 1%, peptone 1%, initial pH 5;
(3) Seed preparation: 3 bacterial slices with the length of 5-7 mm are inoculated in a seed culture medium, and liquid fermentation is carried out for 12-48 hours at the temperature of 25-32 ℃;
(4) Alkali-treating the straw carrier matrix, adding a liquid culture medium, and sterilizing at 121 ℃ for 20min to obtain a solid culture medium;
(5) Inoculating seed liquid into the solid culture medium, controlling the water content to be 55% -80% by inoculating 0.5ml of seeds per gram of straw carrier matrix, and performing aerobic fermentation at 25 ℃ -32 ℃ for 5-7 days to obtain the birch tube-peeling fungus decomposing inoculant.
4. The straw-degrading soil-decomposing inoculant of claim 1, wherein the bacillus subtilis-decomposing inoculant is prepared by the following steps:
(1) Seed culture medium: 1% of peptone, 0.5% of yeast extract, 1% of NaCL and natural pH;
(2) Liquid medium: glucose 0.5%, peptone 1%, yeast extract 0.5%, naCL1% and natural pH;
(3) Seed preparation: b, selecting a bacillus subtilis colony 3 ring, inoculating the bacillus subtilis colony to seed liquid, and carrying out liquid fermentation for 6-36 hours at 37 ℃ by a shaking table at 150 rpm;
(4) Alkali-treating the straw carrier matrix, adding a liquid culture medium, and sterilizing at 121 ℃ for 20min to obtain a solid culture medium;
(5) Inoculating seed liquid into the solid culture medium, controlling the water content to be 55% -80% by inoculating 0.5ml of seeds per gram of straw carrier matrix, and performing aerobic fermentation for 24-48 hours at 37 ℃ to obtain the bacillus subtilis decomposed microbial inoculum.
5. The straw-degrading soil-decomposing inoculant of claim 1, wherein the straw-degrading inoculant is prepared by the following steps:
(1) Seed culture medium: 1% of peptone, 0.5% of yeast extract, 1% of NaCL and natural pH;
(2) Liquid medium: glucose 0.5%, peptone 1%, yeast extract 0.5%, naCL1% and natural pH;
(3) Seed preparation: b, selecting a bacillus subtilis colony 3 ring, inoculating the bacillus subtilis colony to seed liquid, and carrying out liquid fermentation for 6-36 hours at 37 ℃ by a shaking table at 150 rpm;
(4) Alkali-treating the straw carrier matrix, adding a liquid culture medium, and sterilizing at 121 ℃ for 20min to obtain a solid culture medium;
(5) Inoculating seed liquid into the solid culture medium, controlling the water content to be 55% -80% by inoculating 0.5ml of seeds per gram of straw carrier matrix, and performing aerobic fermentation for 24-48 hours at 37 ℃ to obtain the bacillus megaterium decomposing inoculant.
6. The straw-degrading soil-decomposing inoculant according to any one of claims 2 to 5, wherein the alkali-treated straw carrier matrix is prepared by the following method: crushing straws, sieving the crushed straws by a 20-60-mesh sieve, adding 0.5% -1.5% NaOH solution, reacting the crushed straws for 0.5-1.5 hours at the solid-liquid ratio of 1:4-1:6 and the solid-liquid ratio of 50-80 ℃, and flushing the crushed straws with distilled water to pH 6 after the reaction is finished.
7. The application of the straw-degrading soil-decomposing inoculant according to any one of claims 1-6 in field straw decomposition.
8. The application of the straw-degrading soil-decomposing inoculant in field decomposition of farmland straw according to claim 7, wherein when the soil-decomposing inoculant is applied to field decomposition of farmland straw, the straw-decomposing inoculant, the bacillus subtilis-decomposing inoculant and the bacillus megaterium-decomposing inoculant are sequentially inoculated in the order of the straw-decomposing inoculant, the bacillus subtilis-decomposing inoculant and the straw-decomposing inoculant, and the inoculation amount is 0.5-1 g inoculant per 100g straw.
9. The application of the straw-degrading soil-decomposing inoculant in field decomposition of farmland straw according to claim 8, wherein the specific mode of applying the straw-degrading soil-decomposing inoculant to field decomposition of farmland straw is as follows:
(1) Inoculating a sclerotium rolfsii decomposing inoculant, decomposing for 5-7 days, and rolling for 1-3 times/day;
(2) Inoculating a decomposed microbial inoculum of the birch tube-peeling fungus, decomposing for 5-7 days, and rolling for 1-3 times/day;
(3) Inoculating bacillus subtilis decomposing inoculant, decomposing for 2-3 days, and rolling for 1-3 times/day;
(4) Inoculating Bacillus megaterium decomposing inoculant, and simultaneously supplementing 1g/100g of urea and decomposing for 20 days.
CN202210839134.7A 2022-07-18 2022-07-18 Soil decomposing inoculant for degrading straw and application thereof Active CN115369044B (en)

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