CN115369044A - Soil-decomposing inoculant for degrading straw and application thereof - Google Patents
Soil-decomposing inoculant for degrading straw and application thereof Download PDFInfo
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- CN115369044A CN115369044A CN202210839134.7A CN202210839134A CN115369044A CN 115369044 A CN115369044 A CN 115369044A CN 202210839134 A CN202210839134 A CN 202210839134A CN 115369044 A CN115369044 A CN 115369044A
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- 239000010902 straw Substances 0.000 title claims abstract description 128
- 239000002054 inoculum Substances 0.000 title claims abstract description 73
- 230000000593 degrading effect Effects 0.000 title claims abstract description 17
- 244000063299 Bacillus subtilis Species 0.000 claims abstract description 45
- 235000014469 Bacillus subtilis Nutrition 0.000 claims abstract description 45
- 239000002068 microbial inoculum Substances 0.000 claims abstract description 44
- 238000000354 decomposition reaction Methods 0.000 claims abstract description 33
- 241000194107 Bacillus megaterium Species 0.000 claims abstract description 24
- 235000018185 Betula X alpestris Nutrition 0.000 claims abstract description 23
- 235000018212 Betula X uliginosa Nutrition 0.000 claims abstract description 23
- 241000894006 Bacteria Species 0.000 claims abstract description 13
- 241000221955 Chaetomium Species 0.000 claims abstract description 13
- 239000004480 active ingredient Substances 0.000 claims abstract description 3
- 239000001963 growth medium Substances 0.000 claims description 67
- 239000011159 matrix material Substances 0.000 claims description 32
- 239000007788 liquid Substances 0.000 claims description 27
- 239000007787 solid Substances 0.000 claims description 25
- 238000009630 liquid culture Methods 0.000 claims description 24
- 230000000813 microbial effect Effects 0.000 claims description 22
- 238000002360 preparation method Methods 0.000 claims description 19
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 19
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 claims description 18
- 239000001888 Peptone Substances 0.000 claims description 18
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- 238000000034 method Methods 0.000 claims description 18
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- 238000011218 seed culture Methods 0.000 claims description 18
- 239000003795 chemical substances by application Substances 0.000 claims description 17
- 238000011081 inoculation Methods 0.000 claims description 17
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- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 claims description 9
- 238000005096 rolling process Methods 0.000 claims description 9
- 230000001580 bacterial effect Effects 0.000 claims description 8
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- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims description 6
- 239000000843 powder Substances 0.000 claims description 6
- 239000000725 suspension Substances 0.000 claims description 6
- 238000006243 chemical reaction Methods 0.000 claims description 4
- 239000012153 distilled water Substances 0.000 claims description 4
- 239000011148 porous material Substances 0.000 claims description 4
- 229920002472 Starch Polymers 0.000 claims description 3
- XSQUKJJJFZCRTK-UHFFFAOYSA-N Urea Chemical compound NC(N)=O XSQUKJJJFZCRTK-UHFFFAOYSA-N 0.000 claims description 3
- 239000004202 carbamide Substances 0.000 claims description 3
- 239000002131 composite material Substances 0.000 claims description 3
- 229910000365 copper sulfate Inorganic materials 0.000 claims description 3
- ARUVKPQLZAKDPS-UHFFFAOYSA-L copper(II) sulfate Chemical compound [Cu+2].[O-][S+2]([O-])([O-])[O-] ARUVKPQLZAKDPS-UHFFFAOYSA-L 0.000 claims description 3
- 238000007873 sieving Methods 0.000 claims description 3
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- 230000015556 catabolic process Effects 0.000 abstract description 10
- 238000006731 degradation reaction Methods 0.000 abstract description 10
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- 235000002017 Zea mays subsp mays Nutrition 0.000 abstract description 4
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- 230000000694 effects Effects 0.000 description 7
- 239000000126 substance Substances 0.000 description 7
- OAICVXFJPJFONN-UHFFFAOYSA-N Phosphorus Chemical compound [P] OAICVXFJPJFONN-UHFFFAOYSA-N 0.000 description 6
- 239000011574 phosphorus Substances 0.000 description 6
- 229910052698 phosphorus Inorganic materials 0.000 description 6
- 230000008569 process Effects 0.000 description 5
- 108090000790 Enzymes Proteins 0.000 description 4
- 102000004190 Enzymes Human genes 0.000 description 4
- 150000001875 compounds Chemical class 0.000 description 4
- 229940088598 enzyme Drugs 0.000 description 4
- 235000015097 nutrients Nutrition 0.000 description 4
- 241000196324 Embryophyta Species 0.000 description 3
- 244000052616 bacterial pathogen Species 0.000 description 3
- 238000011161 development Methods 0.000 description 3
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- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 2
- 241001112741 Bacillaceae Species 0.000 description 2
- 241000193833 Bacillales Species 0.000 description 2
- 241000221198 Basidiomycota Species 0.000 description 2
- OYPRJOBELJOOCE-UHFFFAOYSA-N Calcium Chemical compound [Ca] OYPRJOBELJOOCE-UHFFFAOYSA-N 0.000 description 2
- CURLTUGMZLYLDI-UHFFFAOYSA-N Carbon dioxide Chemical compound O=C=O CURLTUGMZLYLDI-UHFFFAOYSA-N 0.000 description 2
- 108010059892 Cellulase Proteins 0.000 description 2
- 241000192125 Firmicutes Species 0.000 description 2
- 241000414067 Inonotus obliquus Species 0.000 description 2
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- 108010059896 Manganese peroxidase Proteins 0.000 description 2
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- 229940106157 cellulase Drugs 0.000 description 2
- 238000000855 fermentation Methods 0.000 description 2
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- 244000000000 soil microbiome Species 0.000 description 2
- 241000894007 species Species 0.000 description 2
- 235000000346 sugar Nutrition 0.000 description 2
- 150000008163 sugars Chemical class 0.000 description 2
- IIZPXYDJLKNOIY-JXPKJXOSSA-N 1-palmitoyl-2-arachidonoyl-sn-glycero-3-phosphocholine Chemical compound CCCCCCCCCCCCCCCC(=O)OC[C@H](COP([O-])(=O)OCC[N+](C)(C)C)OC(=O)CCC\C=C/C\C=C/C\C=C/C\C=C/CCCCC IIZPXYDJLKNOIY-JXPKJXOSSA-N 0.000 description 1
- TVEXGJYMHHTVKP-UHFFFAOYSA-N 6-oxabicyclo[3.2.1]oct-3-en-7-one Chemical compound C1C2C(=O)OC1C=CC2 TVEXGJYMHHTVKP-UHFFFAOYSA-N 0.000 description 1
- 241000186361 Actinobacteria <class> Species 0.000 description 1
- 241000222518 Agaricus Species 0.000 description 1
- 235000001674 Agaricus brunnescens Nutrition 0.000 description 1
- 241000002053 Arcopilus cupreus Species 0.000 description 1
- 235000003932 Betula Nutrition 0.000 description 1
- 241000219429 Betula Species 0.000 description 1
- 239000002028 Biomass Substances 0.000 description 1
- BHPQYMZQTOCNFJ-UHFFFAOYSA-N Calcium cation Chemical compound [Ca+2] BHPQYMZQTOCNFJ-UHFFFAOYSA-N 0.000 description 1
- 241001515917 Chaetomium globosum Species 0.000 description 1
- 241001290175 Coriolopsis trogii Species 0.000 description 1
- 241000588724 Escherichia coli Species 0.000 description 1
- 241000123330 Fomes fomentarius Species 0.000 description 1
- 241000159512 Geotrichum Species 0.000 description 1
- 241000143459 Hirsutella Species 0.000 description 1
- FYYHWMGAXLPEAU-UHFFFAOYSA-N Magnesium Chemical compound [Mg] FYYHWMGAXLPEAU-UHFFFAOYSA-N 0.000 description 1
- PWHULOQIROXLJO-UHFFFAOYSA-N Manganese Chemical compound [Mn] PWHULOQIROXLJO-UHFFFAOYSA-N 0.000 description 1
- ZOKXTWBITQBERF-UHFFFAOYSA-N Molybdenum Chemical compound [Mo] ZOKXTWBITQBERF-UHFFFAOYSA-N 0.000 description 1
- BPQQTUXANYXVAA-UHFFFAOYSA-N Orthosilicate Chemical compound [O-][Si]([O-])([O-])[O-] BPQQTUXANYXVAA-UHFFFAOYSA-N 0.000 description 1
- 108090000854 Oxidoreductases Proteins 0.000 description 1
- 102000004316 Oxidoreductases Human genes 0.000 description 1
- 229910019142 PO4 Inorganic materials 0.000 description 1
- 241000123196 Piptoporus betulinus Species 0.000 description 1
- 241000222341 Polyporaceae Species 0.000 description 1
- 241000219000 Populus Species 0.000 description 1
- ZLMJMSJWJFRBEC-UHFFFAOYSA-N Potassium Chemical compound [K] ZLMJMSJWJFRBEC-UHFFFAOYSA-N 0.000 description 1
- 241000589180 Rhizobium Species 0.000 description 1
- 241000124033 Salix Species 0.000 description 1
- 235000016383 Zea mays subsp huehuetenangensis Nutrition 0.000 description 1
- 230000002411 adverse Effects 0.000 description 1
- 229910001424 calcium ion Inorganic materials 0.000 description 1
- -1 carbohydrate compounds Chemical class 0.000 description 1
- 235000014633 carbohydrates Nutrition 0.000 description 1
- 239000001569 carbon dioxide Substances 0.000 description 1
- 229910002092 carbon dioxide Inorganic materials 0.000 description 1
- 238000013329 compounding Methods 0.000 description 1
- 239000013078 crystal Substances 0.000 description 1
- 238000002425 crystallisation Methods 0.000 description 1
- 230000008025 crystallization Effects 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 238000001035 drying Methods 0.000 description 1
- 230000007613 environmental effect Effects 0.000 description 1
- 238000003912 environmental pollution Methods 0.000 description 1
- 239000003337 fertilizer Substances 0.000 description 1
- 238000011010 flushing procedure Methods 0.000 description 1
- 239000000446 fuel Substances 0.000 description 1
- 239000005431 greenhouse gas Substances 0.000 description 1
- 238000003306 harvesting Methods 0.000 description 1
- 229940059442 hemicellulase Drugs 0.000 description 1
- 108010002430 hemicellulase Proteins 0.000 description 1
- 230000003301 hydrolyzing effect Effects 0.000 description 1
- 125000002887 hydroxy group Chemical group [H]O* 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 238000009776 industrial production Methods 0.000 description 1
- 239000003112 inhibitor Substances 0.000 description 1
- 229910052816 inorganic phosphate Inorganic materials 0.000 description 1
- 230000003993 interaction Effects 0.000 description 1
- 239000000787 lecithin Substances 0.000 description 1
- 229940067606 lecithin Drugs 0.000 description 1
- 235000010445 lecithin Nutrition 0.000 description 1
- 239000011777 magnesium Substances 0.000 description 1
- 229910052749 magnesium Inorganic materials 0.000 description 1
- 230000014759 maintenance of location Effects 0.000 description 1
- 235000009973 maize Nutrition 0.000 description 1
- 229910052748 manganese Inorganic materials 0.000 description 1
- 239000011572 manganese Substances 0.000 description 1
- 230000007246 mechanism Effects 0.000 description 1
- 239000002609 medium Substances 0.000 description 1
- 238000009629 microbiological culture Methods 0.000 description 1
- 229910052750 molybdenum Inorganic materials 0.000 description 1
- 239000011733 molybdenum Substances 0.000 description 1
- 150000002772 monosaccharides Chemical class 0.000 description 1
- 229910052757 nitrogen Inorganic materials 0.000 description 1
- 230000003647 oxidation Effects 0.000 description 1
- 238000007254 oxidation reaction Methods 0.000 description 1
- 230000001590 oxidative effect Effects 0.000 description 1
- 244000045947 parasite Species 0.000 description 1
- NBIIXXVUZAFLBC-UHFFFAOYSA-K phosphate Chemical compound [O-]P([O-])([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-K 0.000 description 1
- 239000010452 phosphate Substances 0.000 description 1
- 230000000243 photosynthetic effect Effects 0.000 description 1
- 239000010908 plant waste Substances 0.000 description 1
- 239000011591 potassium Substances 0.000 description 1
- 229910052700 potassium Inorganic materials 0.000 description 1
- 238000004321 preservation Methods 0.000 description 1
- 230000001737 promoting effect Effects 0.000 description 1
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- 239000011573 trace mineral Substances 0.000 description 1
- 235000013619 trace mineral Nutrition 0.000 description 1
- 238000009423 ventilation Methods 0.000 description 1
- 239000002699 waste material Substances 0.000 description 1
- 239000002023 wood Substances 0.000 description 1
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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
- 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
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- 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
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- 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
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- 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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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
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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
- C12N1/20—Bacteria; Culture media therefor
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- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
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- C12R2001/11—Bacillus megaterium
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- C12R2001/07—Bacillus
- C12R2001/125—Bacillus subtilis ; Hay bacillus; Grass bacillus
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- Y02W—CLIMATE CHANGE MITIGATION TECHNOLOGIES RELATED TO WASTEWATER TREATMENT OR WASTE MANAGEMENT
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Abstract
The invention discloses a soil-decomposing inoculant for degrading straws and application thereof. The active ingredients of the soil-decomposing inoculant for degrading straws comprise: the bacillus subtilis comprises a chaetomium fortunei rotten microbial inoculum, a birch tube stripping bacteria rotten microbial inoculum, a bacillus subtilis rotten microbial inoculum and a phosphorus-removing bacillus megaterium rotten microbial inoculum. The soil-decomposing inoculant can be applied to field decomposition of corn straws, rice straw straws, wheat straws and sorghum straws, and the straw degradation rate can reach 60-80%.
Description
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. The straw is the residual product after the crops are harvested, and is also an important organic resource of a farmland ecosystem. China is a big agricultural country, various crop straw resources are very rich, with the development of rural economy and the solution of rural energy problems, straws are no longer used as main fuel, the phenomenon of centralized burning is increasingly prominent in the harvest season, and adverse effects are brought to the environmental air quality. In recent years, how to utilize the straws becomes a research hotspot, wherein straw returning is the first choice, because the straw returning can return nutrients to soil, increase soil organic matters and improve soil quality.
The main components of the straw comprise 30-35% of cellulose, 25-30% of hemicellulose and 20-25% of lignin, and the components need to be 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 has more and more attention in agriculture, particularly straw decomposition at present because the microorganism plays an important role in straw decomposition.
The straw decomposing microbial inoculum is an agricultural microbial inoculum developed from the straw decomposing microbial inoculum, and effective bacteria contained in the agricultural microbial inoculum are viable bacteria preparations processed and prepared after industrial production and propagation. The agricultural microbial agent comprises rhizobium microbial agent, azotobacterium microbial agent, phosphate-solubilizing microbial agent, silicate microbial agent, photosynthetic microbial agent, organic material decomposition agent, growth-promoting microbial agent, mycorrhizal microbial agent, bioremediation microbial agent and the like. The straw decomposing microbial inoculum is one of organic material decomposing microbial 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 massive propagation of the straw decomposing inoculant can effectively decompose crop straws into small organic or inorganic components and return the small organic or inorganic components to soil, so that the improvement of the organic content of the soil is facilitated, and meanwhile, major elements such as nitrogen, phosphorus and potassium and medium and trace elements such as calcium, magnesium, manganese and molybdenum required by the growth of crops are released. The organic matters completely decomposed return to the soil, so that the soil granular structure can be effectively improved, the functions of soil ventilation, fertilizer retention and water retention are improved, heat and a certain amount of carbon dioxide can be generated, the growth environment of plants is improved, and the growth of crops is promoted.
However, due to the complex, tough and heterogeneous straw components, the degradation enzyme systems required by the straws of different crops in the straw decomposition process may be different, and microbial communities have obvious succession phenomena in different periods of straw degradation. Therefore, the existing straw decomposition microbial inoculum has defects in the aspects of applicability and stage property aiming at different crop straws, and influences the degradation effect of the crop straws.
Disclosure of Invention
Based on this, it is necessary to provide a soil-decomposing inoculant which has universality for different crop straws and can meet the requirements of different degradation stages, aiming at the problems of low degradation efficiency and long degradation period of the existing straw-decomposing inoculant for the crop straws.
A soil-decomposing inoculant for degrading straws is a composite inoculant, and active ingredients of the soil-decomposing inoculant comprise: the bacillus subtilis comprises a chaetomium fortunei rotten microbial inoculum, a birch tube stripping bacteria rotten microbial inoculum, a bacillus subtilis rotten microbial inoculum and a phosphorus-removing bacillus megaterium rotten microbial inoculum.
In one embodiment, the content of spore suspension of the Geotrichum rigidum rotten inoculant is 10 5 ～10 7 The spore suspension of the birch tube-peeling bacteria rotten inoculant has the content of 10 per ml 5 ～10 7 The bacillus subtilis decomposing inoculant cfu content is 10 5 ～10 8 The bacillus megaterium phosphate-solubilizing mature microbial inoculum cfu content is 10 5 ～10 8 One per ml.
In one embodiment, the chaetomium fortunei decomposing inoculant is prepared by the following method:
(1) Seed culture medium: 1% of glucose, 1% of malt extract powder and 5% of initial pH;
(2) Liquid culture medium: 0.5% of glucose, 1% of starch, 1% of peptone and 0.005% of copper sulfate, and the initial pH is 5;
(3) Seed preparation: 3 bacterial slices with the diameter of 5-7 mm are inoculated in a seed culture medium and are fermented in a liquid state for 12-48 h at the temperature of 25-32 ℃;
(4) Treating straw carrier matrix with alkali, adding liquid culture medium, and sterilizing at 121 deg.C for 20min to obtain solid culture medium;
(5) Inoculating the seed solution into the solid culture medium, wherein the inoculation amount of the seed solution is 0.5 ml/g of straw carrier matrix, controlling the water content to be 55-80%, and carrying out aerobic fermentation at 25-32 ℃ for 5-7 days to obtain the coarse hirsute pore fungus rotten inoculant.
In one embodiment, the birch tube-peeling fungus-rotting mature microbial inoculum is prepared by the following method:
(1) Seed culture medium: 1% of glucose, 1% of malt extract powder and an initial pH value of 5;
(2) Liquid culture medium: glucose 1%, peptone 1%, initial pH 5;
(3) Seed preparation: 3 bacterial slices with the diameter of 5-7 mm are inoculated in a seed culture medium and are fermented in a liquid state for 12-48 h at the temperature of 25-32 ℃;
(4) Treating straw carrier matrix with alkali, adding liquid culture medium, and sterilizing at 121 deg.C for 20min to obtain solid culture medium;
(5) Inoculating the seed solution into the solid culture medium, wherein the inoculation amount of the seed solution is 0.5 ml/g straw carrier matrix, controlling the water content to be 55-80%, and carrying out aerobic fermentation at 25-32 ℃ for 5-7 days to obtain the birch tube-peeling hole fungus-rotting mature microbial inoculum.
In one embodiment, the bacillus subtilis mature microbial inoculum is prepared by the following method:
(1) Seed culture medium: peptone 1%, yeast extract 0.5%, naCL1%, pH is natural;
(2) Liquid culture medium: 0.5% of glucose, 1% of peptone, 0.5% of yeast extract, 1% of NaCL and natural pH;
(3) Seed preparation: selecting 3 rings of bacillus subtilis colony, inoculating the bacillus subtilis colony in seed liquid, shaking the seed liquid at 37 ℃ and 150rpm, and fermenting the bacillus subtilis colony for 6-36 hours in a liquid state;
(4) Treating straw carrier matrix with alkali, adding liquid culture medium, and sterilizing at 121 deg.C for 20min to obtain solid culture medium;
(5) Inoculating the seed solution into the solid culture medium, wherein the inoculation amount of the seed solution is 0.5ml per gram of straw carrier matrix, controlling the water content of 55-80%, and carrying out aerobic fermentation at 37 ℃ for 24-48 h to obtain the bacillus subtilis maturity fungicide.
In one embodiment, the phosphate-solubilizing bacillus megaterium decomposing inoculant is prepared by the following method:
(1) Seed culture medium: peptone 1%, yeast extract 0.5%, naCL1%, pH is natural;
(2) Liquid culture medium: 0.5% of glucose, 1% of peptone, 0.5% of yeast extract, 1% of NaCL and natural pH;
(3) Seed preparation: selecting 3 rings of bacillus subtilis colony, inoculating the bacillus subtilis colony in seed liquid, shaking the seed liquid at 37 ℃ and 150rpm, and fermenting the bacillus subtilis colony for 6-36 hours in a liquid state;
(4) Treating straw carrier matrix with alkali, adding liquid culture medium, and sterilizing at 121 deg.C for 20min to obtain solid culture medium;
(5) Inoculating the seed solution into the solid culture medium, wherein the inoculation amount of the seed solution is 0.5ml per gram of straw carrier matrix, controlling the water content to be 55-80%, and carrying out aerobic fermentation at 37 ℃ for 24-48 h to obtain the phosphate-solubilizing bacillus megaterium decomposing inoculant.
In one embodiment, the alkali-treated straw carrier matrix is prepared by the following method: crushing straw, sieving with 20-60 mesh sieve, adding 0.5-1.5% concentration NaOH solution in the solid-liquid ratio of 1:4-1:6 at 50-80 deg.c for 0.5-1.5 hr, and flushing with distilled water to pH 6 after the reaction.
The invention also provides application of the soil decomposition microbial inoculum for degrading the straws in field decomposition of the straws of the farmland.
In one embodiment, when the soil decomposition microbial inoculum is applied to field decomposition of farmland straws, the soil decomposition microbial inoculum is inoculated by the chaetomium griffithii decomposition microbial inoculum, the birch tube-peeling bacteria decomposition microbial inoculum, the bacillus subtilis decomposition microbial inoculum and the phosphorus-dissolving bacillus megaterium decomposition microbial inoculum sequentially, wherein the inoculation amount is 0.5-1 g of microbial inoculum per 100g of straws.
In one embodiment, the specific mode of applying the soil decomposition microbial inoculum to field decomposition of farmland straws is as follows:
(1) Inoculating the hard-bristle coarse-capped fungus rotten microbial inoculum, rotting for 5-7 days, and rolling for 1-3 times/day;
(2) Inoculating a birch tube-peeling hole bacterium rotten microbial inoculum, rotting 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 per day;
(4) Inoculating a phosphate-solubilizing bacillus megaterium decomposing inoculant, and simultaneously supplementing 1g/100g of urea to the straws, and decomposing for 20 days.
The straws of the invention refer to corn straws, rice straws, wheat straws and sorghum straws.
The basic aspects of the microorganism species to which the present invention relates are as follows:
in the invention, the adopted chaetomium globosum (Fusallia trogii), the Betula betulinus (Piptoporus betulinus), the Bacillus subtilis (Bacillus subtilis) and the Bacillus megaterium (Bacillus megaterium) are purchased from the China general microbiological culture collection center.
Fomes fomentarius (Funalia trogii): belongs to basidiomycete, mushroom, polyporales, polyporaceae and hirsutella, is a common species in north, is a white rot fungus growing on branches or stumps of broad-leaved trees such as poplar, willow and the like, can efficiently decompose lignin and simultaneously degrade cellulose and hemicellulose.
Birch tube-peeling bacteria (pittoporus betulinus): also called as inonotus obliquus and inonotus obliquus, belonging to basidiomycota, agaricus, polyporales and decorticating fungi, common species in northeast, fruiting bodies are formed in summer and autumn, mostly occur on living standing and falling birch trees, mainly decompose cellulose and hemicellulose, and simultaneously modify lignin, so that brown rot caused by wood can be quickly degraded.
Bacillus subtilis (Bacillus subtilis): belongs to the phylum of firmicutes, the class of bacilli, the order of bacillales, the family of bacillaceae and the genus of bacillus, is a common soil bacterium, is widely distributed in soil and putrefactive organisms, and has strong resistance.
Bacillus megaterium phosphate solubilizing (Bacillus megaterium): belongs to the phylum of firmicutes, the class of bacilli, the order of bacillales, the family of bacillaceae and the genus of bacillus, is a common soil bacterium, can decompose soil phosphide and effectively solve the problem of soil salinization.
The invention starts from the intrinsic structure of the chemical composition of the straws, simulates the mechanism of natural decomposition of the straws by microorganisms, adopts the thought of continuous and sequential decomposition, and purposefully compounds four microorganisms according to the physical and chemical properties of the straws to construct the straw decomposition microbial inoculum. Wherein the chaetomium fortunei can secrete oxidases such as laccase and manganese peroxidase, and oxidize lignin to form small molecular compounds; the birch tube-peeling bacteria can secrete hydrolytic enzymes such as cellulase and hemicellulase, decompose cellulose and hemicellulose, oxidize lignin and improve the content of lignin hydroxyl; the bacillus subtilis has the effects of growing rapidly and regulating the microbial flora in soil, and the organic matters are quickly utilized and the composition of the microbial flora in soil is improved; the phosphate-solubilizing bacillus megaterium has a phosphate-solubilizing function, has an obvious decomposition effect on lecithin and organic phosphorus and inorganic phosphorus which cannot be directly absorbed and utilized by plants in soil, can convert the phosphorus which is difficult to be absorbed and utilized by the plants into an absorbable and utilizable form, and can improve the soil fertility. The four microorganisms generate synergistic action in the composite microbial inoculum, so that the effect of quickly decomposing the straws can be realized, and the soil fertility are improved.
The four methods are used for respectively preparing the chaetomium fortunei hole-removing bacterial decomposing inoculant, the birch tube-peeling bacterial decomposing inoculant, the bacillus subtilis decomposing inoculant and the phosphorus-removing bacillus megaterium decomposing inoculant, the microbial nutrition requirement is low, the culture medium is cheap, common and easy to obtain, the environmental pollution is not generated, the microbial growth speed is high, and the culture condition is simple.
The carrier matrix adopted by the invention has the advantages of wide raw material source, low price, conversion preparation of the carrier matrix and waste recycling; on the other hand, the straw used as a carrier has strong biodegradability, is a farmland crop, can be quickly decomposed under natural conditions, and cannot generate secondary pollution to farmland soil. The method adopts the alkaline pretreatment to prepare the carrier matrix, has simple and convenient operation and low production cost; the specific surface area of the obtained carrier matrix is obviously improved, more microbial agents can be attached, the interaction between the microbes and the carrier matrix is tighter, and the microbes can quickly provide nutrition for the microbial agents after entering the soil, so that the microbes quickly grow, and the influence of severe environment on the microbes is relieved; in addition, compared with a liquid microbial inoculum, the solid carrier has better preservation effect, is easier to transport and the microorganism is easier to grow, and accords with the current new concept of green and clean development.
According to the invention, the inoculation sequence of the chaetomium fortunei rotten inoculant, the birch tube-peeling inoculant, the bacillus subtilis rotten inoculant and the bacillus megaterium phosphate-solubilizing inoculant is adopted in sequence, because the chaetomium fortunei can rapidly utilize cellulose and hemicellulose as nutrients to decompose the substances to generate monosaccharide, laccase, manganese peroxidase and the like generated in the process can perform oxidative fracture on lignin, so that the lignin is decomposed into small molecular compounds, and meanwhile, a cellulose crystallization area is further exposed. The birch tube-peeling bacteria can secrete cellulase and the like, the enzyme further decomposes the exposed cellulose crystal region, and simultaneously performs oxidation modification on the lignin micromolecule substance to further hydroxylate the lignin, so that the water solubility is improved. The micromolecular carbohydrate compounds generated by decomposing the two bacteria can be further used as nutrient substances for the growth and utilization of the bacillus subtilis and the phosphate-solubilizing bacillus megaterium, the two bacilli can grow rapidly to consume the substances, meanwhile, a small amount of micromolecular inhibitor is generated in the growth process of the bacillus subtilis to further inhibit the growth of harmful microorganisms such as soil parasites and escherichia coli, on the other hand, the phosphate-solubilizing bacillus can decompose inorganic phosphate radicals to promote the inorganic phosphorus in soil to be hydrolyzed, and the calcium ions are dissolved to promote the organic phosphorus and calcium to be absorbed.
By applying the soil decomposition microbial inoculum to field decomposition of farmland straws, the straw degradation rate can reach 60-80%.
Compared with the prior art, the invention has the following advantages:
(1) According to the soil-decomposing inoculant disclosed by the invention, aiming at the intrinsic characteristics of straws, the straw components are quickly decomposed by different compounding modes, different components are utilized in a targeted manner, cellulose and hemicellulose are mainly decomposed into micromolecular sugars to provide nutrition for other microorganisms, and other microorganisms grow by utilizing the micromolecular sugars, so that the effects of improving the composition of soil microorganisms and improving the soil nutrients are realized.
(2) In the process of degrading straws 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 soil-decomposing inoculant can be used as soil-decomposing substances to further improve the soil fertility.
(3) The carrier matrix used by the invention is derived from crop residues, has wide sources and low price, can not generate new pollution, has better carrier adsorption performance, and microorganisms can grow on the carrier more easily.
Detailed Description
The present invention will be described in detail with reference to the following embodiments in order to make the aforementioned objects, features and advantages of the invention more comprehensible. In the following description, numerous specific details are set forth in order to provide a thorough understanding of the present invention. This invention may, however, be embodied in many different forms and should not be construed as limited to the embodiments set forth herein.
Example 1 preparation of soil-decomposing inoculant
1-1 preparation of alkali-treated straw carrier substrate: crushing straws, sieving the crushed straws with a 40-mesh sieve, adding 1% NaOH solution into the straws at a solid-to-liquid ratio of 1:5 and 65 ℃, reacting for 1h, and washing the straws with distilled water to reach the pH value of 6 after the reaction is finished.
1-2, preparing a chaetomium fortunei rotten inoculant:
(1) Seed culture medium: 1% of glucose, 1% of malt extract powder and an initial pH value of 5;
(2) Liquid culture medium: 0.5% of glucose, 1% of starch, 1% of peptone and 0.005% of copper sulfate, and the initial pH is 5;
(3) Seed preparation: inoculating 36 mm bacterial slices to a seed culture medium, and performing liquid fermentation for 30 hours at 28 ℃;
(4) Treating straw carrier matrix with alkali, adding liquid culture medium, and sterilizing at 121 deg.C for 20min to obtain solid culture medium;
(5) Inoculating the seed solution into the solid culture medium, wherein the inoculation amount of the seeds is 0.5 ml/g straw carrier matrix, controlling the water content at 70%, and carrying out aerobic fermentation at 28 ℃ for 6 days to obtain the chaetomium fortunei rotten inoculant.
1-3 preparation of birch tube-peeling fungus rotten mature microbial inoculum:
(1) Seed culture medium: 1% of glucose, 1% of malt extract powder and an initial pH value of 5;
(2) Liquid culture medium: glucose 1%, peptone 1%, initial pH 5;
(3) Seed preparation: inoculating 36 mm bacterial sheets to a seed culture medium, and performing liquid fermentation for 30h at 28 ℃;
(4) Treating straw carrier matrix with alkali, adding liquid culture medium, and sterilizing at 121 deg.C for 20min to obtain solid culture medium;
(5) Inoculating the seed solution into the solid culture medium, wherein the inoculation amount of the seeds is 0.5 ml/g straw carrier matrix, controlling the water content at 70%, and carrying out aerobic fermentation at 28 ℃ for 6 days to obtain the birch tube-peeling hole fungus rotten inoculant.
1-4 preparation of Bacillus subtilis decomposing inoculant:
(1) Seed culture medium: peptone 1%, yeast extract 0.5%, naCL1%, pH is natural;
(2) Liquid culture medium: 0.5% of glucose, 1% of peptone, 0.5% of yeast extract, 1% of NaCL and natural pH;
(3) Seed preparation: picking 3 rings of bacillus subtilis colonies, inoculating the bacillus subtilis colonies in seed liquid, and fermenting the bacillus subtilis colonies in a liquid state for 21 hours in a shaking table at 37 ℃ and 150 rpm;
(4) Treating straw carrier matrix with alkali, adding liquid culture medium, and sterilizing at 121 deg.C for 20min to obtain solid culture medium;
(5) Inoculating the seed solution into the solid culture medium, wherein the inoculation amount of the seed solution is 0.5ml per gram of straw carrier matrix, controlling the water content at 70%, and carrying out aerobic fermentation at 37 ℃ for 36h to obtain the bacillus subtilis decomposing inoculant.
1-5 preparation of a phosphate-solubilizing bacillus megaterium decomposing inoculant:
(1) Seed culture medium: peptone 1%, yeast extract 0.5%, naCL1%, pH is natural;
(2) Liquid culture medium: 0.5% of glucose, 1% of peptone, 0.5% of yeast extract, 1% of NaCL and natural pH;
(3) Seed preparation: picking 3 rings of bacillus subtilis colony, inoculating the bacillus subtilis colony in seed liquid, shaking the seed liquid at 37 ℃ and 150rpm, and fermenting the bacillus subtilis colony for 21 hours in a liquid state;
(4) Treating straw carrier matrix with alkali, adding liquid culture medium, and sterilizing at 121 deg.C for 20min to obtain solid culture medium;
(5) Inoculating the seed solution into the solid culture medium, wherein the inoculation amount of the seed solution is 0.5ml per gram of straw carrier matrix, controlling the water content at 70%, and carrying out aerobic fermentation at 37 ℃ for 36h to obtain the phosphate-solubilizing bacillus megaterium decomposing inoculant.
Example 2 field decomposition test of Farmland straw
Pulverizing and scattering the overground part of the harvested maize straw, controlling the water content of 55-80%, and sequentially inoculating chaetomium fortunei and chaetomium cupreum inoculant (the spore suspension content is 4.5 multiplied by 10) 5 One/ml), birch tube-peeling fungus rotten fungus agent (spore suspension content is 6.5X 10) 6 One/ml), bacillus subtilis decomposing inoculant (cfu content is 2.8 multiplied by 10) 7 One/ml), phosphate-solubilizing bacillus megaterium decomposing inoculant (the cfu content is 4.9 multiplied by 10) 6 One/ml), the inoculation amount is 0.75g of microbial inoculum per 100g of straws.
The specific operation steps are as follows:
(1) Inoculating the hard-bristle coarse-capped fungus rotten microbial inoculum, rotting for 5-7 days, and rolling for 1-3 times/day;
(2) Inoculating a birch tube-peeling hole fungus rotten microbial inoculum, rotting 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 per day;
(4) Inoculating a phosphate-solubilizing bacillus megaterium decomposing inoculant, and simultaneously supplementing 1g/100g of urea to the straws, and decomposing for 20 days.
Collecting the decomposed straw, washing with distilled water, removing microbial mycelium on the surface of the substrate as much as possible, putting the substrate into a 63 ℃ oven for drying until the weight is constant, 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 Representing the initial mass of the straw, w f Representing the quality of the straw after decomposition.
The loss rate of a specific component in the straw before and after the straw is decomposed is calculated by the following formula:
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 straws, and Xf representing the content of the certain component in the straws after being decomposed.
The degradation rate of the corn straws is 77 percent through the determination of the method.
The technical features of the embodiments described above may be arbitrarily combined, and for the sake of brevity, all possible combinations of the technical features in the embodiments described above are not described, but should be considered as being within the scope of the present specification as long as there is no contradiction between the combinations of the technical features.
The above-mentioned embodiments only express several embodiments of the present invention, and the description thereof is more specific and detailed, but not construed as limiting the scope of the invention. It should be noted that, for a person skilled in the art, several variations and modifications can be made without departing from the inventive concept, which falls within the scope of the present invention. Therefore, the protection scope of the present patent shall be subject to the appended claims.
Claims (10)
1. The soil-decomposing microbial inoculum for degrading the straws is characterized by being a composite microbial inoculum, and comprising the following active ingredients: the bacillus subtilis strain comprises a chaetomium fortunei rotten microbial agent, a birch tube-peeling bacterium rotten microbial agent, a bacillus subtilis rotten microbial agent and a phosphorus-removing bacillus megaterium rotten microbial agent.
2. According to claim 1The soil-decomposing inoculant for degrading straws is characterized in that the spore suspension content of the hard-bristle coarse-cover-pore-fungus-decomposing inoculant is 10 5 ～10 7 The content of the spore suspension of the birch tube-peeling tube-hole fungus rotten mature microbial inoculum is 10 per ml 5 ～10 7 The bacillus subtilis decomposing inoculant cfu content is 10 5 ～10 8 The content of the decomposed microbial inoculum cfu of the phosphorus-dissolving bacillus megaterium in each ml is 10 5 ～10 8 Each/ml.
3. The straw-degrading soil-decomposing inoculant according to claim 1 or 2, wherein the chaetomium fortunei inoculant is prepared by the following method:
(1) Seed culture medium: 1% of glucose, 1% of malt extract powder and an initial pH value of 5;
(2) Liquid culture medium: 0.5% of glucose, 1% of starch, 1% of peptone and 0.005% of copper sulfate, and the initial pH is 5;
(3) Seed preparation: 3 bacterial slices with the diameter of 5-7 mm are inoculated in a seed culture medium and fermented in a liquid state for 12-48 h at the temperature of 25-32 ℃;
(4) Treating straw carrier matrix with alkali, adding liquid culture medium, and sterilizing at 121 deg.C for 20min to obtain solid culture medium;
(5) Inoculating the seed solution into the solid culture medium, wherein the inoculation amount of the seed solution is 0.5 ml/g of straw carrier matrix, controlling the water content to be 55-80%, and carrying out aerobic fermentation at 25-32 ℃ for 5-7 days to obtain the coarse hirsute pore fungus rotten inoculant.
4. The soil-decomposing inoculant for degrading straw according to claim 1 or 2, wherein the birch tube-peeling inoculant is prepared by the following method:
(1) Seed culture medium: 1% of glucose, 1% of malt extract powder and an initial pH value of 5;
(2) Liquid culture medium: glucose 1%, peptone 1%, initial pH 5;
(3) Seed preparation: 3 bacterial slices with the diameter of 5-7 mm are inoculated in a seed culture medium and fermented in a liquid state for 12-48 h at the temperature of 25-32 ℃;
(4) Treating straw carrier matrix with alkali, adding liquid culture medium, and sterilizing at 121 deg.C for 20min to obtain solid culture medium;
(5) Inoculating the seed solution into the solid culture medium, wherein the inoculation amount of the seed solution is 0.5 ml/g straw carrier matrix, controlling the water content to be 55-80%, and carrying out aerobic fermentation at 25-32 ℃ for 5-7 days to obtain the birch tube-peeling hole fungus-rotting mature microbial inoculum.
5. The soil-decomposing inoculant for degrading straw according to claim 1 or 2, wherein the bacillus subtilis-decomposing inoculant is prepared by the following method:
(1) Seed culture medium: peptone 1%, yeast extract 0.5%, naCL1%, pH is natural;
(2) Liquid culture medium: 0.5% of glucose, 1% of peptone, 0.5% of yeast extract, 1% of NaCL and natural pH;
(3) Seed preparation: selecting 3 rings of bacillus subtilis colony, inoculating the bacillus subtilis colony in seed liquid, shaking the seed liquid at 37 ℃ and 150rpm, and fermenting the bacillus subtilis colony for 6-36 hours in a liquid state;
(4) Treating straw carrier matrix with alkali, adding liquid culture medium, and sterilizing at 121 deg.C for 20min to obtain solid culture medium;
(5) Inoculating the seed solution into the solid culture medium, wherein the inoculation amount of the seed solution is 0.5ml per gram of straw carrier matrix, controlling the water content of 55-80%, and carrying out aerobic fermentation at 37 ℃ for 24-48 h to obtain the bacillus subtilis maturity fungicide.
6. The soil-decomposing inoculant for degrading straw according to claim 1 or 2, wherein the Bacillus megaterium phosphate-solubilizing decomposing inoculant is prepared by the following method:
(1) Seed culture medium: peptone 1%, yeast extract 0.5%, naCL1%, pH is natural;
(2) Liquid culture medium: 0.5% of glucose, 1% of peptone, 0.5% of yeast extract, 1% of NaCL and natural pH;
(3) Seed preparation: selecting 3 rings of bacillus subtilis colony, inoculating the bacillus subtilis colony in seed liquid, shaking the seed liquid at 37 ℃ and 150rpm, and fermenting the bacillus subtilis colony for 6-36 hours in a liquid state;
(4) Treating straw carrier matrix with alkali, adding liquid culture medium, and sterilizing at 121 deg.C for 20min to obtain solid culture medium;
(5) Inoculating the seed solution into the solid culture medium, wherein the inoculation amount of the seed solution is 0.5ml per gram of straw carrier matrix, controlling the water content to be 55-80%, and carrying out aerobic fermentation at 37 ℃ for 24-48 h to obtain the phosphate-solubilizing bacillus megaterium decomposing inoculant.
7. The soil-decomposing inoculant for degrading straw according to any one of claims 3 to 6, wherein the alkali-treated straw carrier matrix is prepared by the following method: crushing the straws, sieving the crushed straws with a 20-60-mesh sieve, adding 0.5-1.5 percent NaOH solution into the straws, reacting the straws for 0.5-1.5 h at the solid-liquid ratio of 1:4-1:6 and the temperature of 50-80 ℃, and washing the straws with distilled water to the pH value of 6 after the reaction is finished.
8. The application of the soil-decomposing inoculant for degrading straw according to any one of claims 1 to 7 in field decomposition of farmland straw.
9. The application of the soil-decomposing inoculant for degrading straw in field decomposition of farmland straw as claimed in claim 8, wherein when the soil-decomposing inoculant is applied to field decomposition of farmland straw, inoculation is performed in the order of the hard-bristle coarse-capped pore fungus-decomposing inoculant, the birch tube-peeling pore fungus-decomposing inoculant, the bacillus subtilis-decomposing inoculant and the phosphorus-dissolving bacillus megaterium-decomposing inoculant, and the inoculation amount is 0.5-1 g inoculant per 100g straw.
10. The application of the soil-decomposing inoculant for degrading straw in field decomposition of farmland straw according to claim 9, wherein the specific way of applying the soil-decomposing inoculant in field decomposition of farmland straw is as follows:
(1) Inoculating the hard-bristle coarse-capped fungus rotten microbial inoculum, rotting for 5-7 days, and rolling for 1-3 times/day;
(2) Inoculating a birch tube-peeling hole bacterium rotten microbial inoculum, rotting 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 per day;
(4) Inoculating a phosphate-solubilizing bacillus megaterium decomposing inoculant, and simultaneously supplementing 1g/100g of urea to the straws, and decomposing for 20 days.
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| VENDULA VALÁŠKOVÁ ET AL.: "Degradation of cellulose and hemicelluloses by the brown rot fungus Piptoporus betulinus-production of extracellular enzymes and characterization of the major cellulases", MICROBIOLOGY, no. 152, pages 3613 - 3622 * |
| 宋丽丽 等: "硬毛粗盖孔菌预处理对玉米秸秆酶水解及组分变化的影响", 中国酿造, vol. 36, no. 2, pages 106 - 110 * |
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