CN115109728A - Straw in-situ decomposition compound microbial agent and preparation method and application thereof - Google Patents
Straw in-situ decomposition compound microbial agent and preparation method and application thereof Download PDFInfo
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- CN115109728A CN115109728A CN202210835454.5A CN202210835454A CN115109728A CN 115109728 A CN115109728 A CN 115109728A CN 202210835454 A CN202210835454 A CN 202210835454A CN 115109728 A CN115109728 A CN 115109728A
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- aspergillus niger
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- 238000011065 in-situ storage Methods 0.000 title claims abstract description 29
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- 238000002360 preparation method Methods 0.000 title claims abstract description 14
- 241000228245 Aspergillus niger Species 0.000 claims abstract description 30
- 241000194108 Bacillus licheniformis Species 0.000 claims abstract description 30
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- 238000000855 fermentation Methods 0.000 claims description 45
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- 239000002609 medium Substances 0.000 claims description 13
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- 229910000019 calcium carbonate Inorganic materials 0.000 claims description 6
- 238000004519 manufacturing process Methods 0.000 claims description 6
- 238000000034 method Methods 0.000 claims description 6
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- NWONKYPBYAMBJT-UHFFFAOYSA-L zinc sulfate Chemical compound [Zn+2].[O-]S([O-])(=O)=O NWONKYPBYAMBJT-UHFFFAOYSA-L 0.000 claims description 4
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- 229910000402 monopotassium phosphate Inorganic materials 0.000 claims description 3
- 235000019796 monopotassium phosphate Nutrition 0.000 claims description 3
- GNSKLFRGEWLPPA-UHFFFAOYSA-M potassium dihydrogen phosphate Chemical compound [K+].OP(O)([O-])=O GNSKLFRGEWLPPA-UHFFFAOYSA-M 0.000 claims description 3
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- 238000001694 spray drying Methods 0.000 claims description 3
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- 229910000357 manganese(II) sulfate Inorganic materials 0.000 claims description 2
- 238000012807 shake-flask culturing Methods 0.000 claims description 2
- 238000007873 sieving Methods 0.000 claims description 2
- 238000004659 sterilization and disinfection Methods 0.000 claims description 2
- 239000002689 soil Substances 0.000 abstract description 13
- 244000005700 microbiome Species 0.000 abstract description 6
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- 241000233866 Fungi Species 0.000 description 3
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- 229940041514 candida albicans extract Drugs 0.000 description 3
- 239000012153 distilled water Substances 0.000 description 3
- 239000011780 sodium chloride Substances 0.000 description 3
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- WQZGKKKJIJFFOK-VFUOTHLCSA-N beta-D-glucose Chemical compound OC[C@H]1O[C@@H](O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-VFUOTHLCSA-N 0.000 description 1
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Abstract
The invention relates to a straw in-situ decomposition compound microbial agent and a preparation method and application thereof, which solve the technical problems of slow decomposition speed and long decomposition time of straws caused by difficult straw returning decomposition under natural conditions and influenced by factors such as stability of straw structures, environmental temperature, soil conditions, microorganisms and the like, and comprises the following strains: bacillus licheniformis (Bacillus licheniformis) is more than or equal to 60 hundred million CFU/g, Rhodotorula mucilaginosa (Rhodotorula mucor) is more than or equal to 2 hundred million CFU/g, and Aspergillus niger (Aspergillus niger) is more than or equal to 20 hundred million CFU/g.
Description
Technical Field
The invention relates to the field of agricultural microorganisms, and in particular relates to a straw in-situ decomposition compound microbial agent and a preparation method and application thereof.
Background
Long-term unreasonable cultivation and intensive land utilization lead to soil degradation problems such as soil layer thinning, organic matter content reduction, structure compaction hardening and the like; however, the straw returning to the field is difficult to decompose under natural conditions, and is influenced by factors such as stability of straw structure, low temperature, soil conditions, microorganisms and the like, so that the decomposition and decomposition speed of the straw is slow, the decomposition time is long, and the planting of next-stubble crops can be seriously influenced under a multiple cropping mode, thereby becoming an obstacle for limiting the straw returning to the field. After the straws are returned to the field, the straws in the early stage are not fermented, so that the phenomena of root hanging, floating and the like of the next crop seedlings are influenced, and the problem of water and nutrient absorption in the field planting process of the crop seedlings is further influenced; during later-period straw fermentation, the straw and crops can compete for nutrients, and the growth and development of the crops are influenced.
Disclosure of Invention
The invention aims to solve the following technical problems in the prior art: the straw returning and decomposing is difficult under natural conditions, and the straw decomposing and decomposing speed is low and the decomposing time is long due to the influence of factors such as the stability of a straw structure, the environmental temperature, the soil condition, microorganisms and the like, so that the invention provides the straw in-situ decomposing compound microbial agent and the preparation method and the application thereof.
Therefore, the invention provides a straw in-situ decomposition compound microbial agent, which contains the following strains: bacillus licheniformis (Bacillus licheniformis) is more than or equal to 60 hundred million CFU/g, Rhodotorula mucilaginosa (Rhodotorula mucor) is more than or equal to 2 hundred million CFU/g, and Aspergillus niger (Aspergillus niger) is more than or equal to 20 hundred million CFU/g.
Preferably, the straw in-situ decomposition compound microbial agent powder comprises the following components in parts by weight: 6-8 parts of bacillus licheniformis, 20-30 parts of aspergillus niger, 1-5 parts of rhodotorula mucilaginosa, 50-60 parts of urea and 10-20 parts of dextrin.
Preferably, the straw in-situ decomposition compound microbial agent powder comprises the following components in parts by weight: 6-8 parts of bacillus licheniformis, 20-30 parts of aspergillus niger, 1-5 parts of rhodotorula mucilaginosa, 50-60 parts of urea, 5-10 parts of sodium carboxymethyl starch and 5-10 parts of light calcium carbonate.
The invention also provides a preparation method of the straw in-situ decomposition compound microbial agent, which comprises the following steps: (1) fermenting the straw in-situ decomposition compound microbial inoculant strain: sequentially carrying out slant culture, shake flask culture, seed culture and fermentation tank culture on bacillus licheniformis, yeast and aspergillus niger strains; counting the viable count of the liquid fermentation of the bacillus licheniformis by more than 100 hundred million CFU/mL, namely judging the termination of fermentation, putting the bacillus licheniformis into a tank, concentrating, adding dextrin, and performing spray drying to obtain full-water-soluble bacillus licheniformis powder; the viable count of yeast liquid fermentation is more than 20 hundred million CFU/mL, namely judging the termination of fermentation, discharging the fermentation product from a tank to obtain a fermentation product, and freeze-drying after ceramic membrane concentration; carrying out Aspergillus niger tray type fermentation, crushing, drying, and sieving with a 100-mesh sieve to obtain fully water-soluble Aspergillus niger powder; the spore count is 100 hundred million CFU/g, and the production is qualified; (2) preparation of the straw in-situ decomposition compound microbial agent: uniformly mixing the bacillus licheniformis, the aspergillus niger and the rhodotorula mucilaginosa with urea and dextrin to obtain a full water-soluble powder microbial decomposing agent; or adding water into the mixture of the bacillus licheniformis, the aspergillus niger and the rhodotorula mucilaginosa, the urea, the sodium carboxymethyl starch and the light calcium carbonate, uniformly mixing and granulating to obtain the solid granular preparation.
Preferably, in the step (1), the production fermentation medium is as follows: and (3) bacillus fermentation medium: 15-20 g/L of corn starch, 20-30 g/L of molasses, 10-20 g/L of bone peptone, KH2PO40.5-1.0 g/L of MgSO4 & 7H2O 0.5-1.0 g/L, MnSO4 & H2O 0.1.1-0.2 g/L and CaCO 30.5-1.0 g/L; the pH value is 7.0-7.2; sterilizing at 121 deg.C for 30 min; yeast liquid fermentation medium: 20-30.0 g/L of molasses, 5-10 g/L of corn starch, 5-10.0 g/L of bone peptone, 2.0-5.0 g/L of yeast powder, 1.0-2.0 g/L of monopotassium phosphate and 0.1-0.5 g/L of magnesium sulfate, and the pH value is 6.0-6.5; sterilizing at 121 ℃ for 30-40 min; aspergillus niger fermentation medium: 25-35 parts of soybean meal, 10-20 parts of corn flour, 10-15 parts of bran, 0.5-1 part of zinc sulfate, 10 parts of water, natural pH, sterilization at 121 ℃ and 30-40 min.
The invention also provides an application of the straw in-situ decomposition compound microbial inoculant as a decomposition agent, wherein the application amount of the straw in-situ decomposition compound microbial inoculant is 1-2 kg/mu or 1-2 kg/(1000kg of straws).
The invention has the following beneficial effects:
the invention develops the composite microbial agent for efficiently decomposing and degrading the straws and the application technology thereof, accelerates the decomposition of the straws and simultaneously ensures the normal growth of crops, so that the in-situ returning of the straws to the field and the improvement of soil organic matters are realized by means of protecting the soil fertility.
Drawings
FIG. 1 is a schematic illustration of the periodic tracking of the microbial inoculant of the present invention during the application of the straw microbial inoculant in saline and alkaline land 1;
FIG. 2 is a schematic diagram of the application of the powdery microbial decomposing agent in corn straw returning decomposition;
FIG. 3 is a granular microbial inoculant of the present invention;
FIG. 4 is a schematic diagram showing the following of the application of the microbial decomposing agent in the northeast region of the present invention.
Detailed Description
The present invention will be further described with reference to the following examples.
The strains are purchased from China general microbiological culture Collection center (CGMCC), wherein the preservation numbers of Bacillus licheniformis (Bacillus licheniformis), Rhodotorula mucilaginosa (Rhodotorula mucosae) and Aspergillus niger (Aspergillus niger) are CGMCC1.6539, CGMCC 2.5541 and CGMCC 3.1380 respectively.
Example 1
Production of high-activity rotten fungi
1. Fermentation of Bacillus licheniformis
Sequentially carrying out slant culture, shake culture, seed culture and fermentation tank culture on the bacillus licheniformis strain. Taking out the preserved bacillus licheniformis strain from a refrigerator at the temperature of-80 ℃, inoculating an LB culture medium, and culturing at the temperature of 37 ℃ for 24 hours; picking single colony from LB culture medium, inoculating to liquid LB culture medium, shake culturing at 200rpm at 37 deg.C for 24 hr; transferring to a 100L fermentation tank filled with seed culture medium, and culturing at 37 deg.C for 20 hr to obtain seed culture solution. Transferring the 10 percent inoculum size of the primary seed culture solution to a fermentation medium 1 ton fermentation tank, and controlling the temperature to be 35 ℃; controlling the dissolved oxygen at 20-30%, rotating at 100-150 rpm, culturing for 18-20 h, and counting viable bacteria by more than 100 hundred million CFU/mL, namely judging that the fermentation is stopped, putting the fermentation tank into a tank, concentrating, adding dextrin, and performing spray drying to obtain the full water-soluble bacillus licheniformis powder.
The preparation process of the culture medium is as follows:
LB culture medium: 5g/L of yeast extract, 10g/L of tryptone and 10g/L of NaCl, and using distilled water to fix the volume to 1.0 liter, wherein the pH value is 7.0-7.2. Adding agar of 15.0g/L into the solid culture medium;
seed culture medium: 10g/L of molasses, 5g/L of yeast extract, 10g/L of tryptone and 5g/L of NaCl;
the formula of the fermentation medium is as follows: corn starch 10g/L, molasses 30g/L, bone peptone 15g/L, KH 2 PO 4 1.0g/L，MgSO 4 ·7H 2 O 0.5g/L，MnSO 4 ·H 2 O 0.2g/L,CaCO 3 1.0gg/L。
Weighing the above materials, adding distilled water to desired volume, sterilizing at 115 deg.C and pH7.2 for 30 min; 20g of agar, i.e., a solid medium, was added to the above medium.
2. Fermentation of Yeast and Aspergillus niger
The yeast and aspergillus niger strains are respectively subjected to slant culture, seed culture and solid fermentation culture in turn. Inoculating the strain into a PYG solid culture medium, and culturing at 28 ℃ for 24-48 h; selecting a single colony, inoculating the single colony in a liquid PYG culture medium, performing shake culture, and culturing at 200rpm and 28 ℃ for 48 hours; obtaining the fungus seed culture solution.
Controlling the temperature of yeast fermentation to be 28 ℃, culturing for 24-32 h, keeping the other operation processes consistent with the fermentation of the bacillus licheniformis, judging that the fermentation product is obtained after the fermentation is stopped when the viable count is more than 20 hundred million CFU/mL, and freeze-drying after ceramic membrane concentration.
Inoculating the aspergillus niger seed liquid into a sterilized and cooled tray filled with a fungus culture medium according to the inoculation amount of 10-20%, inoculating the tray into a solid fermentation chamber, uniformly mixing, adjusting the water content to 50% +/-5%, performing tray fermentation at the ambient temperature of 28 +/-2 ℃, and culturing for 3-5 days to obtain a fermentation product; when the microbial decomposition agent powder is produced, the microbial decomposition agent powder is crushed and dried in advance, and is sieved by a 100-mesh sieve to obtain the fully water-soluble aspergillus niger powder. The spore count is 100 hundred million CFU/g, and the production is qualified.
The preparation process of the culture medium is as follows:
PYG medium: 10.0g/L of peptone, 5.0g/L of yeast extract, 1.0g/L of glucose, distilled water with constant volume pH of 6.0-6.5, sterilizing at 115 ℃ for 30 min; adding 20g of agar into the culture medium to obtain a solid culture medium;
yeast liquid fermentation medium: 30.0g/L of molasses, 10.0g/L of bone peptone, 5.0g/L of yeast powder, 1.0g/L of monopotassium phosphate and 0.5g/L of magnesium sulfate, and the pH value is 6.0-6.5;
aspergillus niger fermentation medium: 30 parts of soybean meal, 15 parts of corn flour (40 meshes), 10 parts of bran, 0.5 part of zinc sulfate and 10 parts of water, and the soybean meal, the corn flour (40 meshes), the bran, the zinc sulfate and the water are sterilized at 121 ℃ and have natural pH.
Example 2
Preparation of dosage form of straw in-situ decomposition compound microbial agent
Uniformly mixing 8 parts of bacillus licheniformis, 30 parts of aspergillus niger, 2 parts of rhodotorula mucilaginosa, 50 parts of urea, 10 parts of dextrin and the like to obtain a full water-soluble powder microbial decomposing agent, wherein the water content is controlled to be less than or equal to 30%; the solid granular formulation is prepared by replacing dextrin in the microbial decomposing agent of the full water-soluble powder with a mixture of equivalent sodium carboxymethyl starch (adhesive) and light calcium carbonate (collapsing agent), wherein the ratio of the sodium carboxymethyl starch to the light calcium carbonate is 1:1, adding a small amount of water, uniformly mixing, granulating (4mm), and drying to control the water content to be less than or equal to 20%.
Example 3
Application of powder microbial decomposition agent in-situ returning of rice straw in saline-alkali soil
Applying the full water-soluble powder microbial decomposing agent to in-situ returning of rice straws planted in Shandong reclamation saline-alkali soil; the test area is 50 mu; smashing rice straws with the stubble left about 50cm to be less than 5-10 cm; the paving thickness is about 5 cm; 1kg of dosage per mu is added with 30 kg of water and sprayed by an unmanned aerial vehicle, and the mixture is uniformly sprayed on the straw; turning the straws into the soil for 10-15 cm; irrigating and flowing into the rice field, and keeping the water level of the rice field about 5cm to submerge the straws. Observations were made every 7 days (table 1).
The application result shows that (as shown in figure 1), the microbial decomposition agent can be applied to returning rice straws in saline-alkali soil to the field, can accelerate the straw decomposition process, does not cause the situation that the microbes compete for nutrients with rice in the decomposition process, has adverse effect on the normal growth of the rice, and simultaneously solves the phenomena of seedling floating and root hanging in the past year.
TABLE 1 straw decomposition conditions during application of straw microbial decomposition agent
Example 4
Application of powder microbial decomposition agent in field returning and decomposition of corn straws
After the corn is picked, the straw amount is relatively large; the large-scale tillage agricultural equipment is difficult to deeply dig all the straws into the soil at one time, and meanwhile, due to the strong seasonality of summer harvesting and summer planting, the plowed straws are not rotten to influence the planting of next-stubble crops. The organic corn planting base in a certain greenhouse in Shandong is selected in the test. Crushing and kneading about 10cm by a corn straw crusher, and paving in a field; uniformly spraying 1kg of powder microbial decomposition agent per 2000 kg of corn straws (the straw yield of about 1 mu of land); in the test, the corn straws are basically rotten in 1 month, the mechanical operation is greatly facilitated, and the corn straws are turned into the soil, so that the corn straws are returned to the field (as shown in figure 2).
Example 5
Application of granular microorganism decomposition agent in rice straw in-situ field recovery in low-temperature region
The solid particle microorganism decomposing inoculant prepared in the above way (as shown in figure 3); the test site is located in Shulan city of Jilin province, the test time is 5 months, the average maximum temperature is about 20 ℃, and the day and night temperature difference is large; selecting 75 mu of rice field, leaving stubble on the straw for 50cm, not crushing the rice straw, uniformly scattering 2kg of microbial decomposition agent particles per mu in the field, then shallow-turning to the underground for 10-20 cm, and covering with water.
The application result shows that the microbial decomposition agent can be applied to returning rice straws in a low-temperature area in northeast China to fields; the granular microbial decomposition agent can be rapidly disintegrated, so that the use is convenient; the high-stubble straws can be thoroughly decomposed without being crushed, so that the labor and mechanical cost is saved; the straw microbial decomposition agent can accelerate the straw decomposition process, can effectively form black humus, and does not affect the field planting, growth and development of rice.
However, the above description is only exemplary of the present invention, and the scope of the present invention should not be limited thereby, and the replacement of the equivalent components or the equivalent changes and modifications made according to the protection scope of the present invention should be covered by the claims of the present invention.
Claims (6)
1. The straw in-situ decomposition compound microbial inoculant is characterized by comprising the following strains: bacillus licheniformis (Bacillus licheniformis) is more than or equal to 60 hundred million CFU/g, Rhodotorula mucilaginosa (Rhodotorula mucoginosa) is more than or equal to 2 hundred million CFU/g, and Aspergillus niger (Aspergillus niger) is more than or equal to 20 hundred million CFU/g.
2. The straw in-situ decomposition compound microbial inoculant according to claim 1, wherein the straw in-situ decomposition compound microbial inoculant powder comprises the following components in parts by weight: 6-8 parts of bacillus licheniformis, 20-30 parts of aspergillus niger, 1-5 parts of rhodotorula mucilaginosa, 50-60 parts of urea and 10-20 parts of dextrin.
3. The straw in-situ decomposition compound microbial inoculant according to claim 1, wherein the straw in-situ decomposition compound microbial inoculant powder comprises the following components in parts by weight: 6-8 parts of bacillus licheniformis, 20-30 parts of aspergillus niger, 1-5 parts of rhodotorula mucilaginosa, 50-60 parts of urea, 5-10 parts of sodium carboxymethyl starch and 5-10 parts of light calcium carbonate.
4. The preparation method of the straw in-situ decomposition compound microbial inoculant as claimed in claim 1, 2 or 3, is characterized by comprising the following steps of:
(1) fermentation of straw in-situ decomposition compound microbial inoculant strain
Sequentially carrying out slant culture, shake flask culture, seed culture and fermentation tank culture on bacillus licheniformis, yeast and aspergillus niger strains;
counting the viable count of the liquid fermentation of the bacillus licheniformis by more than 100 hundred million CFU/mL, namely judging the termination of fermentation, putting the bacillus licheniformis into a tank, concentrating, adding dextrin, and performing spray drying to obtain full-water-soluble bacillus licheniformis powder;
the viable count of yeast liquid fermentation is more than 20 hundred million CFU/mL, namely judging the termination of fermentation, discharging the fermentation product from a tank to obtain a fermentation product, and freeze-drying after ceramic membrane concentration;
carrying out fermentation in a tray type by aspergillus niger, crushing, drying, and sieving by a 100-mesh sieve to obtain fully water-soluble aspergillus niger powder; the spore count is 100 hundred million CFU/g, and the production is qualified;
(2) preparation of compound microbial agent for straw in-situ decomposition
Uniformly mixing the bacillus licheniformis, the aspergillus niger and the rhodotorula mucilaginosa with urea and dextrin to obtain a full water-soluble powder microbial decomposing agent;
or adding water into the mixture of the bacillus licheniformis, the aspergillus niger and the rhodotorula mucilaginosa, the urea, the sodium carboxymethyl starch and the light calcium carbonate, uniformly mixing and granulating to obtain the solid granular preparation.
5. The method for preparing the straw in-situ decomposition composite microbial inoculant powder according to claim 4, wherein in the step (1), the production fermentation medium is as follows:
and (3) bacillus fermentation culture medium: 15-20 g/L of corn starch, 20-30 g/L of molasses, 10-20 g/L of bone peptone, KH2PO40.5-1.0 g/L, 0.5-1.0 g/L of MgSO4 & 7H2O 0.5, 0.1-0.2 g/L of MnSO4 & H2O 0.1, and 30.5-1.0 g/L of CaCO; the pH value is 7.0-7.2; sterilizing at 121 deg.C for 30 min;
yeast liquid fermentation medium: 20-30.0 g/L of molasses, 5-10 g/L of corn starch, 5-10.0 g/L of bone peptone, 2.0-5.0 g/L of yeast powder, 1.0-2.0 g/L of monopotassium phosphate and 0.1-0.5 g/L of magnesium sulfate, and the pH value is 6.0-6.5; sterilizing at 121 ℃ for 30-40 min;
aspergillus niger fermentation medium: 25-35 parts of soybean meal, 10-20 parts of corn flour, 10-15 parts of bran, 0.5-1 part of zinc sulfate, 10 parts of water, natural pH, sterilization at 121 ℃, and 30-40 min.
6. The application of the straw in-situ decomposition compound microbial inoculant as a decomposition agent according to the claim 1, 2 or 3, wherein the application amount of the straw in-situ decomposition compound microbial inoculant is 1-2 kg/mu or 1-2 kg/(1000kg of straws).
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