CN114164151A - Phosphate-solubilizing complex microbial inoculant and application thereof - Google Patents
Phosphate-solubilizing complex microbial inoculant and application thereof Download PDFInfo
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- CN114164151A CN114164151A CN202111464799.6A CN202111464799A CN114164151A CN 114164151 A CN114164151 A CN 114164151A CN 202111464799 A CN202111464799 A CN 202111464799A CN 114164151 A CN114164151 A CN 114164151A
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P3/00—Preparation of elements or inorganic compounds except carbon dioxide
Abstract
The invention relates to a phosphate solubilizing composite microbial inoculum which comprises the following components in percentage by volume: b, bacillus subtilis: 3% -4%; bacillus thuringiensis: 8% -9%; b, bacillus amyloliquefaciens: 1% -2%; streptomyces hygroscopicus: 3% -5%; the balance of water. The mass percentage concentration of the bacillus subtilis is 6-8%. The mass percentage concentration of the bacillus thuringiensis is 5-8%. The mass percentage concentration of the streptomyces ahygroscopicus is 3-5%. The mass percentage concentration of the bacillus amyloliquefaciens is 2-4%. The invention improves the utilization efficiency of phosphorus in the ferro-phosphorus tailings, increases the content of available phosphorus and promotes the effective absorption of plants.
Description
Technical Field
The invention relates to the technical field of ferro-phosphorus tailing treatment, and particularly relates to a phosphate solubilizing composite microbial inoculum and application thereof.
Background
Phosphorite is a non-renewable resource and an irreplaceable resource. Although the reserves of the phosphorite resources in China are rich, the directly utilized high-grade resources are few, and about half of the phosphorite P2O5The grade is below 19 percent, and a large amount of middle-low grade phosphate ores can be more economically utilized after being concentrated by mineral separation. After phosphate ore resources are separated to obtain phosphate concentrate, most of generated tailings are stockpiled as wastes. About 0.44t of tailings are produced for every 1t of phosphate concentrate produced. The generation and accumulation of the phosphate tailings not only cause the loss of phosphorus and other useful resources, but also occupy the land and bring great harm to the ecological environment.
The comprehensive utilization rate of phosphorus tailings in China is low, phosphate ores can generate a large amount of tailings in flotation, the tailings are stacked in a tailing pond for a long time, and in some areas, the tailings are even directly stacked in the open air, so that the environmental pollution is serious. At present, dilute acid leaching method is mainly adopted for secondary recovery of phosphorus tailings, and the method has high cost and low phosphorus recovery.
In order to solve the problem of resource waste and the problems of environment pollution, land resource occupation and the like caused by the resource waste, the secondary recycling of the phosphorus tailings is very important. According to the related data, the phosphorus tailings recovered by the biological fermentation method have low cost and relatively high recovery rate.
Disclosure of Invention
The invention aims to solve the technical problem of providing a phosphate-solubilizing composite microbial inoculum to improve the effective phosphorus content in ferrophosphorus tailings.
In order to achieve the purpose, the invention adopts the following technical scheme:
a phosphate solubilizing complex microbial inoculum comprises the following components in percentage by volume: b, bacillus subtilis: 3% -4%; bacillus thuringiensis: 8% -9%; b, bacillus amyloliquefaciens: 1% -2%; streptomyces hygroscopicus: 3% -5%; the balance of water.
The mass percentage concentration of the bacillus subtilis is 6-8%.
The mass percentage concentration of the bacillus thuringiensis is 5-8%.
The mass percentage concentration of the streptomyces ahygroscopicus is 3-5%.
The mass percentage concentration of the bacillus amyloliquefaciens is 2-4%.
In the phosphate-solubilizing complex microbial inoculum, the effective viable count of bacillus subtilis and bacillus amyloliquefaciens is not less than 1000 hundred million spores/g, the effective viable count of bacillus thuringiensis is not less than 100 hundred million spores/g, and the effective viable count of streptomyces hygroscopicus is not less than 2000 hundred million spores/g.
The application of the phosphate-solubilizing composite microbial inoculum in phosphate tailings is 1-2 percent of the inoculation amount.
Compared with the prior art, the invention has the beneficial effects that:
the invention improves the utilization efficiency of phosphorus in the ferro-phosphorus tailings, increases the content of available phosphorus and promotes the effective absorption of plants.
The phosphate-solubilizing complex microbial inoculum secretes a large amount of phosphatase, phytase and organic acid to the outside of cells in the growth, metabolism, reproduction and process. The bacillus amyloliquefaciens decomposes starch, provides energy substances for the growth of the bacillus subtilis, promotes the bacillus subtilis to grow in a large quantity, consumes oxygen and simultaneously generates phosphorus-containing substances including phospholipids and phosphates (alpha-amylase, protease, lipase and cellulase) and the like, does not absorb streptomyces hygroscopicus to inhibit the growth of other fungi, and provides a sufficient non-competitive environment for the growth of the bacillus subtilis. On one hand, phosphatase and phytase fully phosphorylate phosphate tailings into soluble phosphorus capable of being absorbed and utilized, on the other hand, organic acid can acidify the tailings environment, and under the condition of low pH, the organic acid is chelated with iron, calcium, aluminum and other ions to release phosphate ions, so that the phosphate is dissolved. Soluble phosphorus enters microbial cells, a small part of the phosphorus is utilized by the microbial cells to continuously produce organic acid, and a large part of the phosphorus is reserved. When the microorganisms complete the primary metabolism, a large amount of soluble phosphorus is released and can be absorbed and utilized by crops.
The ferro-phosphorus tailings contain mineral phosphorus which cannot be directly absorbed by plants, and the plants can absorb and utilize available phosphorus, so that the content of the available phosphorus in the ferro-phosphorus tailings is extremely low. In order to fully utilize the phosphorus in the phosphate tailings, the phosphorus-solubilizing compound microbial inoculum is used for changing the total phosphorus in the phosphate tailings into available phosphorus which can be absorbed by plants, and the available phosphorus exists as soluble phosphorus. According to different inoculation ratios, the activity of the phosphatase and the phytase in the tailing culture environment is improved, so that more soluble phosphorus is generated. The increase of the effective phosphorus content indicates that the phosphate-solubilizing composite microbial inoculum has better phosphate-solubilizing effect.
Detailed Description
The invention is further illustrated by the following examples:
the following examples describe the invention in detail. These examples are merely illustrative of the best embodiments of the present invention and do not limit the scope of the invention.
In the embodiment, the effective viable count of the bacillus subtilis and the bacillus amyloliquefaciens in the phosphate-solubilizing composite microbial agent is not less than 1000 hundred million spores/g, the effective viable count of the bacillus thuringiensis is not less than 100 hundred million spores/g, and the effective viable count of the streptomyces hygroscopicus is not less than 2000 hundred million spores/g.
Example 1
The phosphate solubilizing composite microbial inoculum comprises the following volume percentages: b, bacillus subtilis: 3 percent; bacillus thuringiensis: 9 percent; b, bacillus amyloliquefaciens: 1 percent; streptomyces hygroscopicus: 4 percent; the rest water;
wherein the mass percent concentration of the bacillus subtilis is 8 percent, the mass percent concentration of the bacillus thuringiensis is 8 percent, and the mass percent concentration of the streptomyces ahygroscopicus is 3 percent; the mass percentage concentration of the bacillus amyloliquefaciens is 2 percent.
Example 1 application of the phosphate-solubilizing complex bacterial agent in phosphate tailings, wherein the phosphate tailings are fine, the content of-0.038 mm size fraction accounts for 90-92%, the content of available phosphorus is 7.33 mg/kg, and the content of total phosphorus is 35.7 g/kg-1。
The phosphorus tailings inorganic phosphorus culture medium comprises the following components: glucose 10 g.L-1、Ca3(PO4)2 10g·L-1、(NH4)SO4 0.5g·L-1、KCl 0.3g·L-1、MgSO4.7H2O 0.3g·L-1、FeSO4.7H2O 0.03g·L-1、MnSO4.7H2O 0.03g·L-1And the pH value is 7.0-7.2.
The phosphate-solubilizing composite microbial inoculum is inoculated into an inorganic phosphate culture medium of phosphate tailings according to the inoculation amount of 1 percent, the phosphate-solubilizing composite microbial inoculum is fully and uniformly mixed, and then the mixture is placed into a constant-temperature biochemical incubator at 37 ℃ for culture for 7 days, and compared with a blank control, the effective phosphate content is increased by 6.5 times.
Example 2
The phosphate solubilizing composite microbial inoculum comprises the following volume percentages: b, bacillus subtilis: 3 percent; bacillus thuringiensis: 8 percent; b, bacillus amyloliquefaciens: 2 percent; streptomyces hygroscopicus: 3 percent; the rest water;
wherein the mass percent concentration of the bacillus subtilis is 8 percent, the mass percent concentration of the bacillus thuringiensis is 8 percent, and the mass percent concentration of the streptomyces ahygroscopicus is 3 percent; the mass percentage concentration of the bacillus amyloliquefaciens is 2 percent.
Example 2 application of the phosphate-solubilizing complex bacterial agent in phosphate tailings, wherein the phosphate tailings are fine, the content of-0.038 mm size fraction accounts for 90-92%, the effective phosphorus content is 7.33 mg/kg, and the total phosphorus content is 35.7 g/kg-1。
The phosphorus tailings inorganic phosphorus culture medium comprises the following components: glucose 10 g.L-1、Ca3(PO4)2 10g·L-1、(NH4)SO4 0.5g·L-1、KCl 0.3g·L-1、MgSO4.7H2O 0.3g·L-1、FeSO4.7H2O 0.03g·L-1、MnSO4.7H2O 0.03g·L-1And the pH value is 7.0-7.2.
The phosphate-solubilizing composite microbial inoculum is inoculated into an inorganic phosphate culture medium of phosphate tailings according to the inoculation amount of 1 percent, the phosphate-solubilizing composite microbial inoculum is fully and uniformly mixed, and then the mixture is placed into a constant-temperature biochemical incubator at 37 ℃ for culture for 7 days, and compared with a blank control, the effective phosphorus content is increased by 6.6 times.
Example 3
The phosphate solubilizing composite microbial inoculum comprises the following volume percentages: b, bacillus subtilis: 4 percent; bacillus thuringiensis: 8 percent; b, bacillus amyloliquefaciens: 1 percent; streptomyces hygroscopicus: 5 percent; the rest water;
wherein the mass percent concentration of the bacillus subtilis is 8 percent, the mass percent concentration of the bacillus thuringiensis is 8 percent, and the mass percent concentration of the streptomyces ahygroscopicus is 3 percent; the mass percentage concentration of the bacillus amyloliquefaciens is 2 percent.
Example 3 application of the phosphate-solubilizing complex bacterial agent in phosphate tailings, wherein the phosphate tailings are fine, the content of-0.038 mm size fraction accounts for 90-92%, the content of available phosphorus is 7.33 mg/kg, and the content of total phosphorus is 35.7 g/kg-1。
The phosphorus tailings inorganic phosphorus culture medium comprises the following components: glucose 10 g.L-1、Ca3(PO4)2 10g·L-1、(NH4)SO4 0.5g·L-1、KCl 0.3g·L-1、MgSO4.7H2O 0.3g·L-1、FeSO4.7H2O 0.03g·L-1、MnSO4.7H2O 0.03g·L-1And the pH value is 7.0-7.2.
The phosphate-solubilizing composite microbial inoculum is inoculated into an inorganic phosphate culture medium of phosphate tailings according to the inoculation amount of 1 percent, the phosphate-solubilizing composite microbial inoculum is fully and uniformly mixed, and then the mixture is placed into a constant-temperature biochemical incubator at 37 ℃ for culture for 7 days, and compared with a blank control, the effective phosphate content is increased by 6.1 times.
Comparative example 1
The phosphate solubilizing composite microbial inoculum comprises the following volume percentages: b, bacillus subtilis: 4 percent; bacillus thuringiensis: 8 percent; b, bacillus amyloliquefaciens: 3 percent; streptomyces hygroscopicus: 4 percent; the rest water;
wherein the mass percent concentration of the bacillus subtilis is 8 percent, the mass percent concentration of the bacillus thuringiensis is 8 percent, and the mass percent concentration of the streptomyces ahygroscopicus is 3 percent; the mass percentage concentration of the bacillus amyloliquefaciens is 2 percent.
The components of the phosphate tailing culture medium are as follows: glucose 10 g.L-1、Ca3(PO4)2 10g·L-1、(NH4)SO4 0.5g·L-1、KCl0.3g·L-1、MgSO4.7H2O 0.3g·L-1、FeSO4.7H2O 0.03g·L-1、MnSO4.7H2O 0.03g·L-1And the pH value is 7.0-7.2.
The phosphate-solubilizing composite microbial inoculum is inoculated into an inorganic phosphate culture medium of phosphate tailings according to the inoculation amount of 1 percent, the phosphate-solubilizing composite microbial inoculum is fully and uniformly mixed and then placed into a constant-temperature biochemical incubator at 37 ℃ for culture for 7 days, and compared with the embodiment 2, the effective phosphorus content is reduced by 36.9 percent.
Comparative example 2
The phosphate solubilizing composite microbial inoculum comprises the following volume percentages: b, bacillus subtilis: 2 percent; bacillus thuringiensis: 9 percent; b, bacillus amyloliquefaciens: 2 percent; streptomyces hygroscopicus: 5 percent; the rest water;
wherein the mass percent concentration of the bacillus subtilis is 8 percent, the mass percent concentration of the bacillus thuringiensis is 8 percent, and the mass percent concentration of the streptomyces ahygroscopicus is 3 percent; the mass percentage concentration of the bacillus amyloliquefaciens is 2 percent.
The components of the phosphate tailing culture medium are as follows: glucose 10 g.L-1、Ca3(PO4)2 10g·L-1、(NH4)SO4 0.5g·L-1、KCl0.3g·L-1、MgSO4.7H2O 0.3g·L-1、FeSO4.7H2O 0.03g·L-1、MnSO4.7H2O 0.03g·L-1And the pH value is 7.0-7.2.
The phosphate-solubilizing composite microbial inoculum is inoculated into an inorganic phosphate culture medium of phosphate tailings according to the inoculation amount of 1 percent, the phosphate-solubilizing composite microbial inoculum is fully and uniformly mixed and then placed into a constant-temperature biochemical incubator at 37 ℃ for culture for 7 days, and compared with the embodiment 2, the effective phosphorus content is reduced by 27.1 percent.
The influence of the phosphate-solubilizing complex inoculant on the content of available phosphorus in tailings is shown in table 1;
table 1:
as shown in Table 1, the available phosphorus content in the tailings increased with the number of days of cultivation, and reached a maximum at the 7 th day of cultivation, followed by a downward trend. As can be seen from the comparative example and the embodiment, the bacillus amyloliquefaciens in the composite microbial inoculum can generate a plurality of alpha-amylases and proteases and has certain bacteriostatic effect. The ratio of the content of the bacillus subtilis to the content of 3 percent can inhibit the activity of bacillus subtilis and bacillus thuringiensis, and the growth of the bacillus subtilis and the bacillus thuringiensis can be limited when the content of the bacillus subtilis to the content of 1 percent is too low. The bacillus subtilis is aerobic bacteria and has higher activity of producing effective phosphorus, the bacillus amyloliquefaciens is anaerobic bacteria, the contents of the bacillus subtilis and the bacillus amyloliquefaciens need to be balanced to have a growth-dependent relationship, otherwise, the side effect of inhibiting the growth of the other party appears, and the streptomyces hygroscopicus plays a role in inhibiting the production of other harmful bacteria and can promote the transformation and absorption of starch. The utilization efficiency of phosphorus in the ferro-phosphorus tailings is improved, the effective phosphorus content is increased, and the effective absorption of plants is promoted.
Claims (7)
1. The phosphate solubilizing composite microbial inoculum is characterized by comprising the following components in percentage by volume: b, bacillus subtilis: 3% -4%; bacillus thuringiensis: 8% -9%; b, bacillus amyloliquefaciens: 1% -2%; streptomyces hygroscopicus: 3% -5%; the balance of water.
2. The phosphate solubilizing complex microbial inoculant according to claim 1, wherein the mass percentage concentration of the bacillus subtilis is 6-8%.
3. The phosphate solubilizing complex bacterial agent as claimed in claim 1, wherein the mass percentage concentration of bacillus thuringiensis is 5-8%.
4. The phosphate solubilizing complex bacterial agent as claimed in claim 1, wherein the mass percentage concentration of the streptomyces hygroscopicus is 3% -5%.
5. The phosphate solubilizing complex bacterial agent as claimed in claim 1, wherein the mass percentage concentration of bacillus amyloliquefaciens is 2-4%.
6. The phosphate-solubilizing complex microbial inoculum according to any one of claims 1 to 5, wherein the effective viable count of the bacillus subtilis and the bacillus amyloliquefaciens in the phosphate-solubilizing complex microbial inoculum is not less than 1000 hundred million spores/g, the effective viable count of the bacillus thuringiensis is not less than 100 hundred million spores/g, and the effective viable count of the streptomyces hygroscopicus is not less than 2000 hundred million spores/g.
7. The use of the phosphate solubilizing complex bacterial agent according to any one of claims 1 to 5 in phosphate tailings, wherein the inoculation amount is 1 to 2 percent.
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