CN111484368A - Solid fermentation production method of microbial fertilizer and solid composite microbial fertilizer - Google Patents
Solid fermentation production method of microbial fertilizer and solid composite microbial fertilizer Download PDFInfo
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- CN111484368A CN111484368A CN202010283878.6A CN202010283878A CN111484368A CN 111484368 A CN111484368 A CN 111484368A CN 202010283878 A CN202010283878 A CN 202010283878A CN 111484368 A CN111484368 A CN 111484368A
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- C—CHEMISTRY; METALLURGY
- C05—FERTILISERS; MANUFACTURE THEREOF
- C05G—MIXTURES OF FERTILISERS COVERED INDIVIDUALLY BY DIFFERENT SUBCLASSES OF CLASS C05; MIXTURES OF ONE OR MORE FERTILISERS WITH MATERIALS NOT HAVING A SPECIFIC FERTILISING ACTIVITY, e.g. PESTICIDES, SOIL-CONDITIONERS, WETTING AGENTS; FERTILISERS CHARACTERISED BY THEIR FORM
- C05G3/00—Mixtures of one or more fertilisers with additives not having a specially fertilising activity
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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
- C05F1/00—Fertilisers made from animal corpses, or parts thereof
- C05F1/005—Fertilisers made from animal corpses, or parts thereof from meat-wastes or from other wastes of animal origin, e.g. skins, hair, hoofs, feathers, blood
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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
- C05F3/00—Fertilisers from human or animal excrements, e.g. manure
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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
- C05F5/00—Fertilisers from distillery wastes, molasses, vinasses, sugar plant or similar wastes or residues, e.g. from waste originating from industrial processing of raw material of agricultural origin or derived products thereof
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
Abstract
The invention relates to the technical field of microbial fertilizer fermentation, and discloses a microbial fertilizer multi-strain solid synergistic fermentation production method and a solid composite microbial fertilizer. The invention uniformly mixes the fermentation bacteria liquid of four microorganisms of bacillus subtilis, bacillus megaterium, silicate bacteria and streptomyces jingyangensis into the solid fermentation culture medium, and ferments to prepare the solid compound microbial fertilizer, thereby increasing the number of viable bacteria in the fermentation product, promoting the further exertion of the potential of functional strains, enhancing the field colonization of the functional strains, improving the yield of agricultural products and improving the quality of the agricultural products.
Description
Technical Field
The invention relates to the technical field of microbial fertilizer fermentation, in particular to a microbial fertilizer multi-strain solid synergistic fermentation production technology taking composite flora solid fermentation as a core.
Background
The microbial fertilizer is a product which is prepared by compounding organic materials which are obtained by taking specific functional microorganisms and animal and plant residues (such as livestock and poultry manure, crop straws and the like) as sources and are subjected to harmless treatment and decomposition and has the effects of the microbial fertilizer and the organic fertilizer. It can raise crop yield, improve crop quality, raise soil fertility, improve soil physical and chemical properties, improve soil micro ecological system, raise soil biological fertility level and reduce plant diseases and pests.
The development of the microbial fertilizer in China goes through three main development stages. The first development stage, the development of rhizobia and azotobacteria, involves rhizobia preparations. The research center of the microbial fertilizer at the stage is to improve the content of main nutrient elements, particularly the content of nitrogen in the soil by utilizing the characteristics of microorganisms. The microbial fertilizer is characterized in that the effective strains in the product are single strains, and the product is called as a first-generation microbial fertilizer (nutrient simplex type). The second development stage has the focus on exerting the characteristics of various nutritional microorganisms and improving the utilization efficiency of nutrients and fertilizers in the soil. According to the principle of chemical compound fertilizer, single nutrition type strains such as azotobacter, phosphate decomposing bacteria and potassium decomposing bacteria are compounded into microbial fertilizer, and a series of nutrient complementary microbial fertilizers (second generation microbial fertilizers) are developed. The key point of the third development stage is to biologically control growth-promoting bacteria and microbial fertilizer strains with soil remediation effect. At this stage, the types and functions of microbial fertilizer products are gradually diversified, and the multi-strain compound biological fertilizer gradually becomes the mainstream. In the aspect of product composition, a series of products are developed, wherein the microbial inoculum is mixed with a carrier material, and various functional microbial inocula are mixed with chemical fertilizers, particularly in the aspect of compounding functional microorganisms, organic nutrients and inorganic nutrients (complete compound fertilizers).
However, in the production of compound fertilizers in the prior art, a mode of preparing microbial fertilizers by fermenting microbial strain liquid, adsorbing the microbial strain liquid with a solid matrix and aging is mostly adopted. Therefore, the adsorption matrix can not be organically combined with the microorganisms, the microbial fertilizer obtained by mixing is unstable in efficiency, and the characteristics of the functional strains can not be fully exerted.
Disclosure of Invention
In view of the above, the invention provides a microbial fertilizer multi-strain solid synergistic fermentation production method taking composite flora solid fermentation as a core and a solid composite microbial fertilizer prepared by the method. The invention ferments and cultures 4 kinds of microorganism zymocyte solutions of Bacillus Subtilis, Bacillus Megaterium, silicate bacteria and Streptomyces jingyansis in a solid fermentation culture medium, which is beneficial to the organic combination of the microorganism and the fermentation substrate, promotes the further exertion of the potential of functional strains, improves the yield of agricultural products and improves the quality of the agricultural products.
In order to achieve the above object, the present invention provides the following technical solutions:
the invention provides a multi-strain solid synergistic fermentation production method of a microbial fertilizer, which comprises the steps of uniformly mixing fermentation bacteria liquid of four microorganisms of bacillus subtilis, bacillus megatherium, silicate bacteria and streptomyces jingyangensis into a solid fermentation culture medium, and carrying out fermentation culture for 5-7 days at 30-37 ℃ to obtain a solid composite microbial fertilizer;
the solid fermentation medium comprises the following four components in parts by weight:
1) 80-85 parts of edible fungus residues, 8-10 parts of cakes and 10-15 parts of bran;
2) 80-85 parts of fermented chicken manure and 15-20 parts of cake dregs;
3) 75-80 parts of fermented cow dung, 8-15 parts of cakes and 10-12 parts of feather powder;
4) 60-65 parts of edible fungus dregs, 5-10 parts of fermented cow dung, 10-15 parts of fermented chicken manure and 8-12 parts of cake dregs.
The inoculum sizes of the four microbial fermentation bacteria liquids are respectively 5-10%.
Preferably, the material fineness of the solid fermentation medium is 70-100 meshes, the material carbon-nitrogen ratio is 25-30, and the pH value is 6.5-7.5.
In the invention, the four kinds of microbial fermentation bacteria liquid are preferably bacillus subtilis and bacillus megaterium mixed fermentation bacteria liquid, silicate bacteria and streptomyces jingyangensis mixed fermentation bacteria liquid.
Preferably, the preparation method of the mixed fermentation bacteria liquid of the bacillus subtilis and the bacillus megaterium comprises the following steps: inoculating the bacillus subtilis liquid and the bacillus megaterium liquid obtained by shake flask culture into a seed tank according to the inoculation amount of 2.8-3.2% of each, and performing mixed fermentation, and culturing for 22-26 hours to obtain the bacillus subtilis and bacillus megaterium mixed fermentation liquid.
Preferably, the preparation method of the silicate bacteria and streptomyces jingyangensis mixed fermentation bacteria liquid comprises the following steps: inoculating the silicate bacteria liquid and the Jingyang streptomycete liquid obtained by shake flask culture into a seed tank according to the inoculation amount of 2.8-3.2% of each liquid, mixing and fermenting, and culturing for 28-32 hours to obtain the silicate bacteria and Jingyang streptomycete mixed fermentation liquid.
In the present invention, the fermented material is preferably stacked in a length of ×, a width of × and a height of 50m × 5m × 0.5.5 m.
Preferably, the initial moisture content of the fermentation material in the present invention is 50% to 55%.
Preferably, the solid fermentation process of the invention is turned and thrown for 1 time every 22-26 h.
The invention also provides the above-mentioned microorganismThe solid compound microbial fertilizer is obtained by a biological fertilizer multi-strain solid synergistic fermentation production method. The preferable solid composite microbial fertilizer has the live bacteria amount of 30-50 hundred million grams-1(ii) a Further preferably, the Bacillus megaterium-containing composition contains 7 to 15 hundred million g of Bacillus megaterium-120-40 hundred million.g of bacillus subtilis-11-3 hundred million g of silicate bacteria-1Jingyang streptomycete 1-3 hundred million, gram-1。
Compared with the prior art, the invention has the beneficial effects that:
the invention provides a multi-strain solid synergistic fermentation production method of a microbial fertilizer, which is characterized in that fermentation bacteria liquid of four microorganisms, namely bacillus subtilis, bacillus megaterium, silicate bacteria and streptomyces jingyangensis, is uniformly mixed into a solid fermentation culture medium and fermented to prepare the solid composite microbial fertilizer. The invention adopts a solid fermentation mode to carry out the synergistic fermentation of the microorganisms and the solid substrate, and the viable bacteria amount of the four microorganisms reaches 30 to 50 hundred million.g-1And the number of viable bacteria in the fermented product is increased. Meanwhile, the solid fermentation mode is applied to carry out the synergistic fermentation of the microorganisms and the solid substrate, which is beneficial to the organic combination of the microorganisms and the fermentation substrate and promotes the further exertion of the potential of the functional strains.
The solid compound microbial fertilizer produced by solid synergistic fermentation has stable fertilizer effect, can enhance the field colonization of functional strains, effectively reduce the occurrence of soil-borne diseases, promote the development of plant root systems by promoting the growth of metabolites, improve the capability of absorbing soil nutrients and the utilization rate of crops to the nutrients, improve the yield of agricultural products and improve the quality of the agricultural products.
Detailed Description
The invention provides a multi-strain solid synergistic fermentation production method of a microbial fertilizer, which increases the number of viable bacteria in a fermentation product, promotes the organic combination of the microorganisms and a fermentation substrate, and promotes the further exertion of the potential of functional strains, and the specific method comprises the following steps: uniformly mixing fermentation bacteria liquid of four microorganisms including Bacillus Subtilis, Bacillus Megaterium, silicate bacteria and Streptomyces jingyansis into a solid fermentation culture medium, and fermenting and culturing for 5-7 days at the temperature of 30-37 ℃ to obtain the solid composite microbial fertilizer;
the solid fermentation medium comprises the following four components in parts by weight:
1) 80-85 parts of edible fungus residues, 8-10 parts of cakes and 10-15 parts of bran;
2) 80-85 parts of fermented chicken manure and 15-20 parts of cake dregs;
3) 75-80 parts of fermented cow dung, 8-15 parts of cakes and 10-12 parts of feather powder;
4) 60-65 parts of edible fungus dregs, 5-10 parts of fermented cow dung, 10-15 parts of fermented chicken manure and 8-12 parts of cake dregs.
In the invention, the inoculum sizes of the four microbial zymophyte liquids are respectively 5-10%.
As an implementable mode, the strain applied in the invention is preferably bacillus subtilis (Bacillus subtilis), and the strain preservation number of the China agricultural microbial strain preservation management center is ACCC 19742; bacillus Megaterium (Bacillus Megaterium) with the strain preservation number of ACCC10011 of China agricultural microorganism strain preservation management center; silicate bacteria (Bacillus mucoginosus), wherein the strain preservation number of the China agricultural microbial strain preservation management center is ACCC 10013; streptomyces jingyanniensis (Streptomyces jingyansis), and the strain preservation number of the China agricultural microbial strain preservation management center is ACCC 40021.
In a preferred embodiment, the present invention uses a mixed fermentation broth of bacillus subtilis and bacillus megaterium, and a mixed fermentation broth of silicate bacteria and streptomyces jingyangensis to perform solid fermentation. The bacillus subtilis and bacillus megaterium mixed fermentation bacterial liquid and the silicate bacteria and the streptomyces jingyangensis are mixed pairwise and do not generate antagonistic action, after pairwise mixing fermentation, the proportion of the two bacterial liquids in each seed tank is kept constant, and further the fertilizer efficiency is stable after the four bacteria are mixed and solid fermented. The invention adopts the form of mixing and fermenting two kinds of bacteria, thereby not only saving fermentation equipment and reducing investment cost, but also shortening fermentation period.
The invention also provides a preparation method of the bacillus subtilis and bacillus megaterium mixed fermentation bacterial liquid and the silicate bacteria and streptomyces jingyangensis mixed fermentation bacterial liquid, which comprises the following steps:
the preparation method of the mixed fermentation bacterial liquid of the bacillus subtilis and the bacillus megaterium comprises the following steps: inoculating the bacillus subtilis liquid and the bacillus megaterium liquid obtained by shake flask culture into a seed tank according to the inoculation amount of 2.8-3.2% of each, and performing mixed fermentation, and culturing for 22-26 hours to obtain the bacillus subtilis and bacillus megaterium mixed fermentation liquid.
The preparation method of the silicate bacteria and streptomyces jingyangensis mixed fermentation bacterial liquid comprises the following steps: inoculating the silicate bacteria liquid and the Jingyang streptomycete liquid obtained by shake flask culture into a seed tank according to the inoculation amount of 2.8-3.2% of each liquid, mixing and fermenting, and culturing for 28-32 hours to obtain the silicate bacteria and Jingyang streptomycete mixed fermentation liquid.
Among them, as a preferred embodiment, the shake flask culture conditions in the present invention are: culturing the bacillus subtilis and the bacillus megaterium at 28-30 ℃ for 18-22h at 180-; culturing silicate bacteria at 28-30 ℃ for 28-32h at 180-; culturing the Jingyang streptomycete at 25-28 ℃ for 40-44h at 180-.
Seeding tank culture conditions: the preferred mixed fermentation bacterial liquid of the bacillus subtilis and the bacillus megaterium is as follows: the culture temperature is 28-32 ℃, the culture rotation speed is 180-200 r/min, and the culture ventilation is 1: 1 (v: v/min); the preferable selection of the mixed fermentation bacterial liquid of silicate bacteria and streptomyces jingyangensis is as follows: the culture temperature is 28-32 ℃, the culture rotation speed is 180-: 1 (v: v/min).
In a preferred embodiment of the present invention, the culture medium for shake flask culture and seeding tank fermentation of four microorganisms is preferably:
and (3) shake flask culture:
(1) liquid culture medium of bacillus subtilis:
potato 200 g
20 g of cane sugar
1000 ml of distilled water
pH 6.0～6.5；
(2) Liquid culture medium of bacillus megaterium:
beef extract 5.0 g
Peptone 5.0 g
Sodium chloride 5.0 g
1000 ml of distilled water
pH 7.2～7.4；
(3) Silicate bacteria liquid culture medium:
sucrose 5 g
Yeast extract 0.8 g
Ammonium sulfate 0.1 g
Magnesium sulfate 0.8 g
K2HPO41 g
Sodium chloride 0.1 g
Calcium carbonate 1 g
1000 ml of distilled water
pH 7.0～7.2；
(4) Jingyang streptomycete liquid culture medium
Soluble starch 20 g
KNO31.0 g
K2HPO40.5 g
MgSO4·7H2O0.5 g
NaCl 0.5 g
FeSO4·7H2O0.01 g
1000 ml of distilled water
pH 7.2～7.4；
Seeding tank culture
(1) The fermentation culture medium of the bacillus subtilis and the bacillus megatherium seed tank comprises the following components in percentage by weight:
0.2 percent of ammonium sulfate
0.286 percent of glucose
0.714% of potato starch
Dipotassium phosphate 0.030%
0.020% of sodium chloride
0.020% of potassium chloride
Magnesium sulfate 0.080%
Manganese sulfate 0.003%
Ferrous sulfate 0.003%
0.300 percent of yeast powder
98.344% of distilled water;
(2) the weight percentages of the silicate bacteria and the fermentation medium of the Jingyang streptomycete strain tank are as follows:
0.5 percent of bean cake powder
0.5 percent of cane sugar
0.5 percent of yeast powder
0.01 percent of ammonium sulfate
Magnesium sulfate 0.08%
Dipotassium phosphate 0.1%
0.01 percent of sodium chloride
0.1 percent of calcium carbonate
Ferrous sulfate 0.001%
98.199% of distilled water.
The material source in the solid fermentation culture medium is not particularly limited, and the known or commercially available raw materials in the prior art can be used for preparing the solid compound microbial fertilizer.
In a preferred embodiment, the fermented chicken manure and the fermented cow manure are obtained by fermenting chicken manure and cow manure in a farm, but the present invention is not particularly limited to the technical means of the present invention.
In a preferred embodiment, the fermented chicken manure has the following technical indexes: water content of 30 +/-5%, organic matter of 45 +/-5%, total nitrogen of 1.5 +/-0.5%, P2O51.7±0.3％、K2O1.9 plus or minus 0.3 percent; the technical indexes of the fermented cow dung are as follows: 35 +/-5% of water content, 45 +/-5% of organic matter, 1.3 +/-0.3% of total nitrogen and P2O51.3±0.3％、K2O 1.6±0.3％。
As an implementable technical scheme, the cakes, the waste feather puffed powder and the bran are not particularly limited, and the known or commercially available raw materials in the prior art can be used in the invention. As a preferred embodiment, the technical indexes of the cakes in the invention are as follows: total nitrogen 4.8 + -0.5%, P2O51.5±0.3％、K2O0.16 +/-0.03%; the technical indexes of the waste feather puffed powder (feather powder) are as follows: total nitrogen 13.3 + -0.3%, P2O50.17±0.03％、K2O0.21 +/-0.03%; the technical indexes of the bran are as follows: the crude protein is more than or equal to 11.0 percent, and the crude fiber is less than 11.0 percentCoarse ash content is less than 6.0%.
As a preferred embodiment, the fineness of the material of the solid fermentation medium in the present invention is 70 to 100 mesh, and more preferably 80 mesh. The invention crushes the materials, is beneficial to uniformly mixing the solid materials and the microbial liquid, can accelerate the reaction speed of fermentation, and is beneficial to the organic combination of the microbial liquid and the adsorption matrix.
In the present invention, the carbon-nitrogen ratio and the pH of the solid material are defined, and as a preferred embodiment, the carbon-nitrogen ratio of the solid material of the present invention is 25 to 30, the pH is 6.5 to 7.5, and the carbon-nitrogen ratio of the solid material of the present invention is further preferably 28, and the pH is 7. The carbon-nitrogen ratio and the pH value of the material defined in the invention ensure the carbon-nitrogen balance of the compost mixture, increase the number of viable bacteria in the fermented product, promote the further exertion of the potential of functional strains and enable the microorganisms to achieve better biological activity.
The solid fermentation can be conventionally stacked, as a preferred embodiment, the fermented materials are stacked according to the length of ×, the width of × and the height of 50m × 5m × 0.5.5 m, and are preferably turned and thrown for 1 time every 22-26h and further preferably once every 24h in the solid fermentation process.
The initial water content of the fermentation material is defined in the invention, and as a preferred embodiment, the initial water content of the fermentation material is 50-55%. The preferred water content of the invention ensures that the material absorbs water, swells and softens, and is beneficial to the decomposition of microorganisms. If the water content is too high, the air permeability of the stack is influenced, and the activity of functional microorganisms is inhibited; if the water content is too low, the activity and the reproduction of microorganisms can be slowed down, and the fermentation decomposition speed is influenced.
The invention also provides a solid compound microbial fertilizer obtained by the microbial fertilizer multi-strain solid synergistic fermentation production method. As a preferred embodiment, the solid composite microbial fertilizer of the present invention has a viable bacteria amount of 30 to 50 hundred million g-1(ii) a More preferably, the Bacillus megaterium-containing food contains 7-15 hundred million g of Bacillus megaterium-120-40 hundred million.g of bacillus subtilis-1Silicate bacteria 1-3Hundred million grams-1Jingyang streptomycete 1-3 hundred million, gram-1。
The solid compound microbial fertilizer produced by the solid synergistic fermentation has stable fertilizer effect, can enhance the field colonization of functional strains, effectively reduce the occurrence of soil-borne diseases, promote the development of plant root systems by promoting the growth of metabolites, improve the capability of absorbing soil nutrients and the utilization rate of crops to the nutrients, improve the yield of agricultural products and improve the quality of the agricultural products.
The technical solutions of the present invention will be described in detail with reference to the following examples, but the following examples should not be construed as limiting the scope of the present invention.
In the following examples of the present invention, the following raw materials and types of microorganisms are used, and those skilled in the art can still appropriately substitute or replace the raw materials and the types of microorganisms according to the source conditions of the raw materials and the microorganisms, which should not be construed as limiting the technical scope of the present invention.
The fermented chicken manure has the following technical indexes: 30.52% of water content, 45.53% of organic matter, 1.44% of total nitrogen and P2O51.70％、K2O 1.92％；
The fermented cow dung has the following technical indexes: 35.23% of water content, 45.19% of organic matter, 1.33% of total nitrogen and P2O51.36％、K2O 1.65％；
Cakes, commercially available, technical indexes: total nitrogen 4.84%, P2O51.51％、K2O 0.16％；
The waste feather puffed powder (feather powder) is sold in the market, and has the technical indexes that: total nitrogen 13.26%, P2O50.17％、K2O0.21％；
And (3) microbial strains: bacillus Subtilis (Bacillus Subtilis), and the strain preservation number of China agricultural microorganism strain preservation management center is ACCC 19742; bacillus Megaterium (Bacillus Megaterium) with the strain preservation number of ACCC10011 of China agricultural microorganism strain preservation management center; silicate bacteria (Bacillus mucoginosus), wherein the strain preservation number of the China agricultural microbial strain preservation management center is ACCC 10013; streptomyces jingyansis (Streptomyces jingyansis), and the strain preservation number of the China agricultural microbial strain preservation management center is ACCC 40021;
bran, commercially available, technical index: the crude protein is more than or equal to 11.0 percent, the crude fiber is less than 11.0 percent, and the crude ash content is less than 6.0 percent.
Example 1
Liquid fermentation of strain
1. And (3) shake flask culture:
(1) culture of Bacillus subtilis liquid
Culture medium: potato (peeled, cooked) 200 g
Sucrose (white sugar or glucose) 20 g
1000 ml of distilled water
pH 6.0～6.5
Injecting 150ml of culture medium into a 500ml triangular flask; transferring the strains on the inclined plane into a corresponding shake flask by using 5ml of sterile water, and culturing for 20h at the temperature of 28-30 ℃ under the condition of 200 revolutions per minute.
(2) Culture of bacillus megatherium liquid
Culture medium: beef extract 5.0 g
Peptone 5.0 g
Sodium chloride 5.0 g
pH 7.2～7.4
1000 ml of distilled water
Injecting 150ml of culture medium into a 500ml triangular flask; transferring the strains on the inclined plane into a corresponding shake flask by using 5ml of sterile water, and culturing for 20h at the temperature of 28-30 ℃ under the condition of 200 revolutions per minute.
(3) Culture of silicate bacteria liquid
Sucrose 5 g
Yeast extract 0.8 g
Ammonium sulfate 0.1 g
Magnesium sulfate 0.8 g
K2HPO41 g
Sodium chloride 0.1 g
Calcium carbonate 1 g
pH7.0～7.2
Injecting 150ml of culture medium into a 500ml triangular flask; transferring the strains on the inclined plane into a corresponding shake flask by using 5ml of sterile water, and culturing for 30 hours at the temperature of 28-30 ℃ under the condition of 200 revolutions per minute.
(4) Culture medium of Jingyang streptomycete
Culture medium: soluble starch 20 g
KNO31.0 g
K2HPO40.5 g
MgSO4·7H2O0.5 g
NaCl 0.5 g
FeSO4·7H2O0.01 g
1000 ml of distilled water
pH7.2～7.4
Injecting 150ml of culture medium into a 500ml triangular flask; transferring the strain on the inclined plane into a corresponding shake flask by using 5ml of sterile water, and culturing for 42h at the temperature of 25-28 ℃ under the condition of 200 revolutions per minute.
2. Seeding tank culture
The preferable various culture media are as follows, wherein the percentages are weight percentages:
(1) fermentation culture of bacillus subtilis and bacillus megaterium in seed tank
Culture medium: 0.2 percent of ammonium sulfate
0.286 percent of glucose
0.714% of potato starch
Dipotassium phosphate 0.030%
0.020% of sodium chloride
0.020% of potassium chloride
Magnesium sulfate 0.080%
Manganese sulfate 0.003%
Ferrous sulfate 0.003%
0.300 percent of yeast powder
98.344 percent of distilled water
The bacillus subtilis liquid and the bacillus megaterium liquid are respectively added into a seed tank according to the inoculation amount of 3 percent and then mixed and fermented; the temperature is 30 ℃, the culture speed is 180-200 r/min, and the culture ventilation is 1: 1 (v: v/min); the culture time is 24 hours, and the bacteria are detected within 20 hours.
(2) Fermenting and culturing silicate bacteria and streptomyces jingyangensis in a seed tank
Culture medium: 0.5 percent of bean cake powder
0.5 percent of cane sugar
0.5 percent of yeast powder
0.01 percent of ammonium sulfate
Magnesium sulfate 0.08%
Dipotassium phosphate 0.1%
0.01 percent of sodium chloride
0.1 percent of calcium carbonate
Ferrous sulfate 0.001%
98.199 percent of distilled water
Adding the silicate bacteria solution and the Jingyang streptomycete solution into a seed tank according to the inoculation amount of 3% of each solution, and then mixing and fermenting; temperature 30 ℃, culture speed 200 r/min, culture ventilation 1: 1 (v: v/min); the culture time is 30 hours, and the bacteria detection is carried out for 28 hours.
Example 2
Solid fermentation of strains:
the solid fermentation medium comprises the following components in parts by weight:
80 parts of edible fungus residues, 8 parts of cakes, 10 parts of bran, 80-mesh fineness of the material, 28 carbon-nitrogen ratio of the material and pH 7;
the two seed tank zymophyte liquids in the example 1 are respectively mixed into the solid fermentation culture medium according to the inoculation amount of 10 percent to ensure that 4 types of bacteria are uniformly mixed, the fermented materials are stacked according to the length of ×, the width of × and the height of 50m × 5m × 0.5.5 m, the initial water content of the raw materials is 55 percent, the fermentation culture temperature is 35 ℃, the fermentation time is 7 days, and the solid composite microbial fertilizer product 1 is obtained after 24 hours of turning and polishing.
The bacterium content and the nutritional indexes in the obtained product are detected, and the results are as follows:
product 1: the effective viable bacteria amount is 50.0 hundred million/gram, wherein the bacillus megaterium amount is 10 hundred million g-1Bacillus subtilis 35 hundred million.g-13 hundred million g of silicate-1Jingyang streptomycete 2 hundred million g-1The number of the mold and the mixed bacteria is less than or equal to 3.0 × 106Per gram, the water content is less than or equal to 30 percent, the fineness is more than or equal to 80 percent, and the pH value is 7.5; the shelf life is 12 months; the dosage form of the product is powder, and the organic matter is more than or equal to 40 percent.
Example 3
The solid fermentation medium comprises the following components in parts by weight:
85 parts of fermented chicken manure, 20 parts of cake meal, 80-mesh material fineness, 25 material carbon-nitrogen ratio and 6.5 pH value.
The microbial zymophyte liquid is prepared according to the method of the example 1, the two seed tank zymophyte liquids are respectively added with 4 types of bacteria according to the inoculation amount of 5 percent to be evenly mixed into the solid fermentation culture medium, the fermented material is stacked according to the length of ×, the width of × and the height of 50m × 5m × 0.5.5 m, the initial water content of the raw material is 50 percent, the fermentation culture temperature is 30 ℃, the fermentation time is 5 days, and the solid composite microbial fertilizer product 2 is obtained after 24 hours of turning and polishing for 1 time.
The bacterium content and the nutritional indexes in the obtained product are detected, and the results are as follows:
product 2: the effective viable bacteria amount is 32.0 hundred million/gram, wherein the bacillus megaterium is 8 hundred million g-1Bacillus subtilis 22 hundred million g-1Silicate bacteria 1 hundred million g-1Jingyang streptomycete 1 hundred million g-1The number of the mold and the mixed bacteria is less than or equal to 3.0 × 106Per gram, the water content is less than or equal to 30 percent, the fineness is more than or equal to 80 percent, and the pH value is 7.5; the shelf life is 12 months; the dosage form of the product is powder, and the organic matter is more than or equal to 40 percent.
Example 4
The solid fermentation medium comprises the following components in parts by weight:
75 parts of fermented cow dung, 15 parts of cakes, 12 parts of feather powder, 80-mesh material fineness, 28 material carbon-nitrogen ratio and 7 pH value.
The microbial zymophyte liquid is prepared according to the method of the example 1, the two seed tank zymophyte liquids are respectively added with 4 types of bacteria according to the inoculation amount of 10 percent to be evenly mixed into the solid fermentation culture medium, the fermented material is stacked according to the length of ×, the width of × and the height of 50m × 5m × 0.5.5 m, the initial water content of the raw material is 55 percent, the fermentation culture temperature is 37 ℃, the turning and polishing are carried out for 1 time in 24 hours, and the fermentation time is 7 days, so that the solid composite microbial fertilizer product 3 is obtained.
The bacterium content and the nutritional indexes in the obtained product are detected, and the results are as follows:
product 3: the effective viable bacteria amount is 45.0 hundred million/gram, wherein the bacillus megaterium is 10 hundred million g-1Bacillus subtilis 30 hundred million g-13 hundred million g of silicate-1Jingyang streptomycete 2 hundred million g-1The number of the mold and the mixed bacteria is less than or equal to 3.0 × 106Per gram, the water content is less than or equal to 30 percent, the fineness is more than or equal to 80 percent, and the pH value is 7.5; the shelf life is 12 months; the dosage form of the product is powder, and the organic matter is more than or equal to 40 percent.
Example 5
The solid fermentation medium comprises the following components in parts by weight:
65 parts of edible fungus dregs, 10 parts of fermented cow dung, 15 parts of fermented chicken manure and 12 parts of cake dregs, and the edible fungus dregs, the fermented cow dung, the fermented chicken manure and the cake dregs are uniformly mixed, the material fineness is 80 meshes, the carbon-nitrogen ratio of the material is 30, and the pH value is 7.5.
The microbial zymophyte liquid is prepared according to the method of the example 1, two seed tank zymophyte liquids are respectively added according to the inoculation amount of 5 percent to evenly mix 4 types of bacteria into the solid fermentation culture medium, the fermented materials are stacked according to the length of ×, the width of × and the height of 50m × 5m × 0.5.5 m, the initial water content of the raw materials is 50 percent, the fermentation culture temperature is 35 ℃, the turning and polishing are carried out for 1 time in 24 hours, and the fermentation time is 6 days, so as to obtain the solid composite microbial fertilizer product 4.
The bacterium content and the nutritional indexes in the obtained product are detected, and the results are as follows:
product 4: the effective viable bacteria amount is 40.0 hundred million/gram, wherein the bacillus megaterium is 8 hundred million g-1Bacillus subtilis 30 hundred million g-11 hundred million g of silicate-1Jingyang streptomycete 1 hundred million g-1The number of the mold and the mixed bacteria is less than or equal to 3.0 × 106Per gram, the water content is less than or equal to 30 percent, the fineness is more than or equal to 80 percent, and the pH value is 7.5; the shelf life is 12 months; the dosage form of the product is powder, and the organic matter is more than or equal to 40 percent.
Example 6
Field test
In order to verify the specific effect of the solid compound microbial fertilizer obtained by the multi-strain solid synergistic fermentation production method, the inventor conducts a test of a plot with the continuous cropping age of more than 10 years on the Shandong Jinxianghu Town, sheep Shanwn Town, fish Shanwn Town and Xinglong Town in 2018-. The microbial fertilizer prepared by the invention is used as a field test demonstration, and a conventional fertilizing mode of farmers and a similar microbial agent sold in the market are used as a contrast.
Wherein the garlic variety applied in the invention is Jinxiang white-skin garlic;
the conventional fertilization mode of farmers is as follows: applying a commercially available product for fertilization, applying 100 kg/mu of a special (18-10-17) compound fertilizer for garlic as a base fertilizer, and applying 20 kg/mu of a special (18-10-17) compound fertilizer for garlic before Qingming festival in the next year;
commercially available microbial agents of the same type: the effective viable count (cfu) is more than or equal to 60 hundred million/g, the microbial strains are Bacillus megatherium (Bacillus megatherium), Bacillus Subtilis (Bacillus Subtilis) and Bacillus polymyxa (Paenibacillus polymyxa), and the organic matter is more than or equal to 40 percent;
the test group is the solid compound microbial fertilizer product 1 prepared in the embodiment 2 of the invention;
the fertilizer application mode of the commercially available similar microbial inoculum and the test group of the invention is that 1 kg is applied during garlic sowing, 1 kg is applied before winter and 1 kg is applied during the green turning period on the basis of conventional fertilizer application, and pesticide is used by half in the whole growth period compared with conventional fertilizer application treatment. The using effect of the plot over 10 years of continuous cropping is compared with the following table:
TABLE 1 comparison of the application effect of the solid compound microbial fertilizer in continuous cropping over 10 years
From the table, it is known that, compared with the conventional fertilization, the garlic yields of the solid compound microbial fertilizer prepared in the example 2 and the commercially available microbial inoculum are increased by 16.2% and 13.51% respectively, which are significantly different from the conventional fertilization; the yield of the young garlic shoots is increased by 3.5 percent and 2.3 percent respectively compared with the yield of the young garlic shoots by conventional fertilization, and the difference between the three treatments is not obvious. The solid compound microbial fertilizer prepared in the embodiment 2 and the microbial inoculum sold in the market have the effect of reducing the root rot disease of garlic, and have an obvious disease-resistant effect compared with the microbial inoculum sold in the market.
As can be seen from the above examples, the present invention utilizes the solid fermentation mode to co-ferment the microorganisms and the solid substrate, and the viable bacteria amount of the four microorganisms reaches 30-50 hundred million.g-1Wherein the bacillus megatherium is 7-15 hundred million g-120-40 hundred million.g of bacillus subtilis-1Silicate bacteria 3-5 hundred million g-1Jingyang streptomycete 1-3 hundred million, gram-1And the number of viable bacteria in the fermented product is increased. Meanwhile, the solid fermentation mode is applied to carry out the synergistic fermentation of the microorganisms and the solid substrate, which is beneficial to the organic combination of the microorganisms and the fermentation substrate and promotes the further exertion of the potential of the functional strains.
The solid compound microbial fertilizer produced by solid synergistic fermentation has stable fertilizer effect, can enhance the field colonization of functional strains, effectively reduce the occurrence of soil-borne diseases, promote the development of plant root systems by promoting the growth of metabolites, improve the capability of absorbing soil nutrients and the utilization rate of crops to the nutrients, improve the yield of agricultural products and improve the quality of the agricultural products.
The foregoing is only a preferred embodiment of the present invention, and it should be noted that, for those skilled in the art, various modifications and decorations can be made without departing from the principle of the present invention, and these modifications and decorations should also be regarded as the protection scope of the present invention.
Claims (10)
1. A multi-strain solid synergistic fermentation production method of a microbial fertilizer is characterized in that fermentation bacteria liquid of four microorganisms, namely bacillus subtilis, bacillus megaterium, silicate bacteria and streptomyces jingyangensis, is uniformly mixed into a solid fermentation culture medium and is fermented and cultured for 5-7 days at the temperature of 30-37 ℃ to obtain a solid composite microbial fertilizer;
the solid fermentation medium comprises any one of the following four components in parts by weight:
1) 80-85 parts of edible fungus residues, 8-10 parts of cakes and 10-15 parts of bran;
2) 80-85 parts of fermented chicken manure and 15-20 parts of cake dregs;
3) 75-80 parts of fermented cow dung, 8-15 parts of cakes and 10-12 parts of feather powder;
4) 60-65 parts of edible fungus dregs, 5-10 parts of fermented cow dung, 10-15 parts of fermented chicken manure and 8-12 parts of cake dregs.
2. The method for producing microbial fertilizer by multi-strain solid synergistic fermentation as claimed in claim 1, wherein the inoculum sizes of the four microbial fermentation broth are 5-10% respectively.
3. The method for producing the microbial fertilizer by the multi-strain solid synergistic fermentation as claimed in claim 1, wherein the fineness of the solid fermentation medium is 70-100 meshes, the carbon-nitrogen ratio of the material is 25-30, and the pH value is 6.5-7.5.
4. The method for producing microbial fertilizer by multi-strain solid synergistic fermentation as claimed in claim 1, wherein the four kinds of microbial fermentation broth are mixed fermentation broth of bacillus subtilis and bacillus megaterium, and mixed fermentation broth of silicate bacteria and streptomyces jingyangensis.
5. The method for producing microbial fertilizer by multi-strain solid synergistic fermentation as claimed in claim 4, wherein the preparation method of the mixed fermentation bacteria liquid of bacillus subtilis and bacillus megaterium comprises the following steps: inoculating the bacillus subtilis liquid and the bacillus megaterium liquid obtained by shake flask culture into a seed tank according to the inoculation amount of 2.8-3.2% of each, and performing mixed fermentation, and culturing for 22-26 hours to obtain the bacillus subtilis and bacillus megaterium mixed fermentation liquid.
6. The method for producing microbial fertilizer by multi-strain solid synergistic fermentation according to claim 4, wherein the preparation method of the silicate bacteria and streptomyces jingyangensis mixed fermentation bacteria liquid comprises the following steps: inoculating the silicate bacteria liquid and the Jingyang streptomycete liquid obtained by shake flask culture into a seed tank according to the inoculation amount of 2.8-3.2% of each liquid, mixing and fermenting, and culturing for 28-32 hours to obtain the silicate bacteria and Jingyang streptomycete mixed fermentation liquid.
7. The method for producing microbial fertilizer by multi-strain solid synergistic fermentation as claimed in claim 1, wherein the initial water content of the fermentation material is 50-55%.
8. The method for producing microbial fertilizer by multi-strain solid synergistic fermentation as claimed in claim 1, wherein the solid fermentation process is carried out by turning over for 1 time every 22-26 h.
9. The solid compound microbial fertilizer obtained by the method for producing the microbial fertilizer by the multi-strain solid synergistic fermentation of claims 1 to 8, wherein the viable bacteria content of the solid compound microbial fertilizer reaches 30 to 50 hundred million grams-1。
10. The solid composite microbial fertilizer according to claim 9, wherein the solid composite microbial fertilizer contains 7-15 hundred million-g of bacillus megaterium-120-40 hundred million.g of bacillus subtilis-11-3 hundred million g of silicate bacteria-1Jingyang streptomycete 1-3 hundred million, gram-1。
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| CN114773127A (en) * | 2022-03-14 | 2022-07-22 | 宁夏地丰生物科技有限公司 | Organic fertilizer fermentation process |
| CN115010553A (en) * | 2022-05-30 | 2022-09-06 | 滨州市京阳生物肥业有限公司 | Soil structure improved bio-organic fertilizer and preparation method thereof |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN114773127A (en) * | 2022-03-14 | 2022-07-22 | 宁夏地丰生物科技有限公司 | Organic fertilizer fermentation process |
| CN115010553A (en) * | 2022-05-30 | 2022-09-06 | 滨州市京阳生物肥业有限公司 | Soil structure improved bio-organic fertilizer and preparation method thereof |
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