CN114656293A - Method for improving survival rate of microorganisms in high inorganic nutrient fertilizer - Google Patents
Method for improving survival rate of microorganisms in high inorganic nutrient fertilizer Download PDFInfo
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- 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
- C05F11/08—Organic fertilisers containing added bacterial cultures, mycelia or the like
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- 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
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Abstract
The invention discloses a method for improving the survival rate of microorganisms in a high inorganic nutrient fertilizer, belonging to the field of biological fertilizers. The method comprises the following steps: (1) mixing the microorganism and a protective agent according to the mass ratio of 1-4:1-5 to prepare a microorganism bacterium agent; (2) adding the microbial agent into the high inorganic nutrient fertilizer in a proportion of 0.1-10 wt%, and then drying or mixing and granulating to obtain the microbial compound fertilizer. The method can effectively improve the survival rate of the bacterial strains in the microbial compound fertilizer in the production and storage processes, the survival rate of the bacterial strains in one-year shelf life reaches 70-91 percent, and the preparation and storage problems of the microbial compound fertilizer are solved to a great extent; the microbial compound fertilizer prepared by the invention can effectively promote the growth of crops, remarkably improve the yield of the crops, reduce the application amount of chemical fertilizers and pesticides, reduce the economic cost, improve the economic benefit and provide technical support for developing green agriculture.
Description
Technical Field
The invention relates to the field of biological fertilizers, in particular to a method for improving the survival rate of microorganisms in a high inorganic nutrient fertilizer.
Background
Yield of agricultural products depends on the interaction of the plant rhizosphere with microorganisms, and rhizosphere conditions determine the health and productivity of the plant. The microorganisms can maintain various trace and macro nutrients in the soil environment to be rich by fixing nitrogen, dissolving or mineralizing phosphate, releasing plant growth regulating factors and degrading organic matters in the soil. The single application of the microbial inoculum can not meet the growth requirement of crops, and the agricultural production mainly depends on the high inorganic nutrient fertilizer, combines the microbes and the high inorganic nutrient fertilizer, provides comprehensive nutrition in the growth process of plants and simultaneously prevents and controls plant diseases and insect pests. The combination of the microorganisms and the high inorganic nutrient fertilizer can promote the nutrient conversion of soil, improve the nutrient utilization rate of the fertilizer, reduce the problems of soil hardening and environmental pollution, increase the yield and income of crops and greatly improve the agricultural economic benefit.
The research of adding microorganisms into the compound fertilizer is carried out, the microbial agent and the organic-inorganic compound fertilizer are mixed and granulated, the survival rate of viable bacteria is only 35-65% after granulation is finished, and the survival rate of strains in shelf storage period after production is not monitored. The current technology of adding microorganisms into high inorganic nutrient fertilizers is difficult to break through, and the content of inorganic nutrients in the microbial organic-inorganic compound fertilizer can only reach about 15%. In the process of mixing and granulating the microorganisms and the organic-inorganic compound fertilizer, a large number of strains die due to high temperature of machine granulation, and high osmotic pressure is formed by inorganic salt in the storage process, so that fatal stress is brought to the microorganisms. Therefore, it is desirable to provide a method for increasing the survival of microorganisms in high inorganic nutrient fertilizers.
Disclosure of Invention
The invention aims to provide a method for improving the survival rate of microorganisms in a high inorganic nutrient fertilizer, which aims to solve the problems in the prior art.
In order to achieve the purpose, the invention provides the following scheme:
the invention provides a method for improving the survival rate of microorganisms in a high inorganic nutrient fertilizer, which comprises the following steps:
(1) mixing the microorganism and a protective agent according to the mass ratio of 1-4:1-5 to prepare a microorganism bacterium agent;
(2) adding the microbial agent into the high inorganic nutrient fertilizer in a proportion of 0.1-10 wt%, and then drying or mixing and granulating to obtain the microbial compound fertilizer.
Further, the protective agent comprises the following components in percentage by weight: 1-28% of vegetable oil, 0.5-26% of glycerin, 1-10% of sorbitol, 1-10% of mannitol and the balance of water.
Further, the high inorganic nutrient fertilizer comprises an inorganic fertilizer, an organic fertilizer and a carrier.
Further, the weight percentage of the inorganic fertilizer in the microbial compound fertilizer is 35-45%, the weight percentage of the organic fertilizer in the microbial compound fertilizer is 13-24%, and the weight percentage of the carrier in the microbial compound fertilizer is 38-42%.
Further, the drying condition is cold air drying at 20-55 ℃ for 4-6 h.
Further, the microorganism comprises bacillus, and the bacillus is prepared by mixing bacillus powder and a protective agent.
The invention also provides a microbial compound fertilizer prepared by the method.
The invention discloses the following technical effects:
the invention provides a method for improving the survival rate of microorganisms in a high inorganic nutrient fertilizer, optimizes the production method of adding the microorganisms into the high inorganic nutrient fertilizer, combines a microbial agent with the high nutrient inorganic fertilizer, and can be mixed with 45% of inorganic nutrient compound fertilizer at most, thereby realizing the preparation of the high microbial activity high inorganic nutrient compound fertilizer.
The protective agent can effectively improve the survival rate of the bacterial strains in the microbial compound fertilizer in the production and storage processes, and the survival rate of the bacterial strains in one-year shelf life is between 70 and 91 percent. Breaks through the technical bottleneck of the microbial compound fertilizer, combines the microbial agent and the high inorganic nutrient fertilizer, ensures the high activity of the strain, and solves the preparation and storage problems of the microbial compound fertilizer to a great extent. In addition, field test results show that the microbial compound fertilizer prepared by the method can effectively promote the growth of crops, remarkably improve the yield of the crops, reduce the application amount of chemical fertilizers and pesticides, reduce the economic cost, improve the economic benefit and provide technical support for developing green agriculture.
Drawings
In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings needed in the embodiments will be briefly described below, and it is obvious that the drawings in the following description are only some embodiments of the present invention, and it is obvious for those skilled in the art to obtain other drawings without creative efforts.
FIG. 1 shows the statistics of the copy number of Bacillus subtilis gene in capsicum soil;
FIG. 2 shows the statistical results of the copy number of the Bacillus subtilis gene in tomato soil.
Detailed Description
Reference will now be made in detail to various exemplary embodiments of the invention, the detailed description should not be construed as limiting the invention but as a more detailed description of certain aspects, features and embodiments of the invention.
It is to be understood that the terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention. Further, for numerical ranges in this disclosure, it is understood that each intervening value, between the upper and lower limit of that range, is also specifically disclosed. Every smaller range between any stated value or intervening value in a stated range and any other stated or intervening value in a stated range is encompassed within the invention. The upper and lower limits of these smaller ranges may independently be included or excluded in the range.
Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Although only preferred methods and materials are described herein, any methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present invention. All documents mentioned in this specification are incorporated by reference herein for the purpose of disclosing and describing the methods and/or materials associated with the documents. In case of conflict with any incorporated document, the present specification will control.
It will be apparent to those skilled in the art that various modifications and variations can be made in the specific embodiments of the present disclosure without departing from the scope or spirit of the disclosure. Other embodiments will be apparent to those skilled in the art from consideration of the specification. The description and examples are intended to be illustrative only.
As used herein, the terms "comprising," "including," "having," "containing," and the like are open-ended terms that mean including, but not limited to.
The methods used in the following examples are conventional methods unless otherwise specified; the reagents used are all commonly available on the market unless otherwise specified.
1 materials and methods
1.1 test materials
Bacterial agent: bacillus species: although bacillus subtilis strains are selected to prepare microbial compound fertilizers in the following examples, other bacillus biocontrol strains are as follows: bacillus megaterium, Bacillus licheniformis, Bacillus amyloliquefaciens and the like can also be used for obtaining the microbial compound fertilizer with high survival rate by adopting the method.
Fertilizer: high inorganic nutrient fertilizer: comprises inorganic fertilizer and organic fertilizer, also called organic-inorganic compound fertilizer (provided by Shizhu agriculture science and technology limited of Hubei);
carrier: fulvic acid;
a protective agent: vegetable oil, glycerol, sorbitol, mannitol;
1.2 preparation of the culture Medium
Luria-Bertani liquid medium (bacterial culture medium): to 950ml of deionized water were added 10g of Tryptone (Tryptone), 5g of Yeast extract (Yeast extract) and 10g of sodium chloride (NaCl), stirred with a glass rod until the solute was dissolved, adjusted to pH 7.2. + -. 0.2 with 5mol/L of NaOH, and then diluted to 1L with deionized water. Sterilizing at 121 deg.C for 20 min.
Luria-Bertani solid medium: the components are the same as Luria-Bertani liquid culture medium, and 15g-20g of agar powder is added after other components are dissolved. Sterilizing at 121 deg.C for 20 min.
Fermentation tank culture medium: 40g/L of soybean meal, 20g/L of corn flour, 15g/L of glucose, 3g/L of dipotassium phosphate, 1.5g/L of monopotassium phosphate and 0.2g/L of yeast extract powder.
1.3 design of the experiment
1.3.1 preparation of Bacillus subtilis powder
The strain was activated with a solid LB plate and inoculated into a 500ml Erlenmeyer flask (liquid content 100ml) for culture for 16 h. Inoculating the bacillus subtilis into a 50L fermentation tank according to the inoculation amount of 1%, fermenting for 60h, and putting into a spray dryer to obtain the bacillus subtilis powder.
1.3.1 protective agent encapsulation
Weighing a proper amount of bacillus powder, and mixing the bacillus powder and a protective agent according to a mass ratio of 1-4:1-5 to prepare the microbial agent. Uniformly coating the microbial agent on the surface of the compound microbial fertilizer according to the addition amount of 0.1-10 wt%, and drying the compound fertilizer coated with the microbial agent in a cold air dryer at 20-55 ℃ for 4-6h to prepare the microbial compound fertilizer, or mixing and granulating the microbial agent and the compound fertilizer in a granulator to prepare the microbial compound fertilizer. The viable count of the microbial compound fertilizer prepared in the following examples is more than or equal to 0.2 hundred million cfu/g.
Example 1
A microbial compound fertilizer comprises the following components in parts by weight: 5 parts of microbial agent, 40 parts of inorganic fertilizer, 13 parts of organic fertilizer and 42 parts of carrier.
The preparation method of the microbial compound fertilizer comprises the following steps:
(1) uniformly mixing bacillus powder and a protective agent in a solid-liquid mass ratio of 1:1 to prepare a microbial agent for later use;
the protective agent comprises 11 wt% of vegetable oil, 8 wt% of glycerin, 8 wt% of sorbitol, 1 wt% of mannitol and the balance of water.
(2) Uniformly spraying the microbial preparation on the surface of the organic-inorganic compound fertilizer according to the proportion.
(3) And (3) putting the compound fertilizer wrapped with the microbial agent into a cold air dryer to dry for 5 hours at 40 ℃ to prepare the microbial compound fertilizer.
Example 2
A microbial compound fertilizer comprises the following components in parts by weight: 4 parts of microbial agent, 40 parts of inorganic fertilizer, 15 parts of organic fertilizer and 41 parts of carrier.
The preparation method of the microbial compound fertilizer comprises the following steps:
(1) uniformly mixing bacillus powder and a protective agent in a solid-liquid mass ratio of 1:2 to prepare a microbial agent for later use;
the protective agent comprises 17 wt% of vegetable oil, 11 wt% of glycerin, 1.6 wt% of sorbitol, 7.5 wt% of mannitol and the balance of water.
(2) The microbial preparation, the organic fertilizer, the inorganic fertilizer and the carrier are put into a granulator according to the proportion to be mixed and granulated, so as to prepare the microbial compound fertilizer.
Example 3
A microbial compound fertilizer comprises the following components in parts by weight: 5 parts of microbial agent, 38 parts of inorganic fertilizer, 18 parts of organic fertilizer and 39 parts of carrier.
The preparation method of the microbial compound fertilizer comprises the following steps:
(1) uniformly mixing bacillus powder and a protective agent in a solid-liquid mass ratio of 2:3 to prepare a microbial agent for later use;
the protective agent comprises 28 wt% of vegetable oil, 4 wt% of glycerol, 5.3 wt% of sorbitol, 6.8 wt% of mannitol and the balance of water.
(2) Uniformly spraying the microbial preparation on the surface of the organic-inorganic compound fertilizer according to the proportion.
(3) And (3) putting the compound fertilizer wrapped with the microbial agent into a cold air dryer to be dried for 4 hours at the temperature of 55 ℃ to prepare the microbial compound fertilizer.
Example 4
A microbial compound fertilizer comprises the following components in parts by weight: 4 parts of microbial agent, 45 parts of inorganic fertilizer, 20 parts of organic fertilizer and 31 parts of carrier.
The preparation method of the microbial compound fertilizer comprises the following steps:
(1) uniformly mixing bacillus powder and a protective agent in a solid-liquid mass ratio of 1:1 to prepare a microbial agent for later use;
the protective agent comprises 1 wt% of vegetable oil, 26 wt% of glycerol, 10 wt% of sorbitol, 1 wt% of mannitol and the balance of water.
(2) Uniformly spraying the microbial preparation on the surface of the organic-inorganic compound fertilizer according to the proportion.
(3) And (3) drying the compound fertilizer wrapped with the microbial agent in a cold air dryer at 20 ℃ for 6 hours to prepare the microbial compound fertilizer.
Example 5
A microbial compound fertilizer comprises the following components in parts by weight: 10 parts of microbial agent, 35 parts of inorganic fertilizer, 16 parts of organic fertilizer and 39 parts of carrier.
The preparation method of the microbial compound fertilizer comprises the following steps:
(1) uniformly mixing bacillus powder and a protective agent in a solid-liquid mass ratio of 4:5 to prepare a microbial agent for later use;
the protective agent comprises 25 wt% of vegetable oil, 0.5 wt% of glycerin, 1 wt% of sorbitol, 10 wt% of mannitol and the balance of water.
(2) Uniformly spraying the microbial preparation on the surface of the organic-inorganic compound fertilizer according to the proportion.
(3) And (3) putting the compound fertilizer wrapped with the microbial agent into a cold air dryer to dry for 5 hours at 40 ℃ to prepare the microbial compound fertilizer.
Example 6
A microbial compound fertilizer comprises the following components in parts by weight: 0.1 part of microbial agent, 37.9 parts of inorganic fertilizer, 24 parts of organic fertilizer and 38 parts of carrier.
The preparation method of the microbial compound fertilizer comprises the following steps:
(1) uniformly mixing bacillus powder and a protective agent in a solid-liquid mass ratio of 1:2 to prepare a microbial agent for later use;
the protective agent comprises 10 wt% of vegetable oil, 11 wt% of glycerin, 4.3 wt% of sorbitol, 5.6 wt% of mannitol and the balance of water.
(2) The microbial preparation, the organic fertilizer, the inorganic fertilizer and the carrier are put into a granulator according to the proportion to be mixed and granulated, so as to prepare the microbial compound fertilizer.
Comparative example 1
The difference from the embodiment 1 is that the microbial agent is prepared by adding 5g of activated carbon, 5g of diatomite, 30g of soluble starch and 6g of glycerol into each liter of bacillus fermentation liquor, and the number of viable bacteria in the microbial agent is not less than 200 hundred million/ml.
Comparative example 2
The difference from example 1 is that the protective agent lacks sorbitol.
Effect example 1
The microbial compound fertilizers prepared in examples 1 to 6 were stored at room temperature for one year, and then the survival rates of the strains in the respective compound fertilizers were counted, and the results are shown in table 1:
TABLE 1
| Group of | Strain survival rate (%) |
| Example 1 | 87.21 |
| Example 2 | 71.25 |
| Example 3 | 91.43 |
| Example 4 | 90.22 |
| Example 5 | 91.28 |
| Example 6 | 76.17 |
| Comparative example 1 | 62.19 |
| Comparative example 2 | 60.24 |
Effect example 2 field test
1 materials and methods
1.1 test conditions and materials
1.1.1 test conditions
The pepper test site is located in the Wen jia village in the autonomous county of the Changyang Tujia family of the province of Hubei, and the tomato test site is located in the general depression in the autonomous county of the Changyang Tujia family of the province of Hubei, with the elevation of 1200-1600 meters, which is a typical mountain climate.
1.1.2 test varieties
Chili jinyou
Tomato Tianxi F1
1.1.3 test Fertilizer varieties
40% (16-8-16) of compound fertilizer (provided by Shizhu agriculture science and technology Co., Ltd., Hubei).
1.2 design of the experiment
The fertilizers of the treatment groups described in Table 2 were obtained by the preparation method of example 4.
The test time is from 5 months to 10 months in 2021, the fertilizer is applied in 5 months to 4 days in 5 months, and the field planting is carried out in 20 days in 5 months. Setting 5 treatments, respectively planting pepper and tomato in two test areas, wherein the total area of 15 cells is 60-70 m2Three replicates of each treatment were performed. The test treatments are shown in Table 2. Planting density of pepper 45223 strains/hm2Planting density of 34422 tomato plants/hm2. After the plants are matured through conventional management, the yield and net profit of each treatment product are respectively counted.
Table 2 test treatments
1.3 sample Collection and measurement
Collecting 0-20 cm plough layer soil samples in each test area by a five-point sampling method at 7-12 days, 8-9 days, 9-12 days and 10-15 days, respectively, standing at-80 ℃ for storage, and extracting DNA solution. A Bacillus subtilis specific primer gyrB (refer to Xie S, Yu H, Wang Q, et al) is selected for qPCR quantification. And (4) counting the copy number of the bacillus subtilis gene in the soil.
1.4 results and analysis
1.4.1 yield of the products of each treatment
TABLE 3 Pepper yield, net profit
TABLE 4 tomato yield, net profit
Tables 3 and 4 show that the microbial organic-inorganic compound fertilizer wrapped by the protective agent can effectively promote the growth of crops, remarkably improve the yield of the crops, reduce the application amount of chemical fertilizers and pesticides, develop green agriculture, reduce economic cost and improve economic benefit.
1.4.2 viable count of Bacillus subtilis in soil
According to the copy number of the bacillus subtilis gene in each treated capsicum soil in figure 1, the protective agent can effectively improve the survival rate of the strain after being applied to the soil, C-FBS-2 added with the protective agent treatment can maintain the microbial agent at an order of magnitude three months ago, the protective agent is not added in the C-FBS-1 treatment to wrap the microbial agent, and the bacterial number is continuously reduced at 60d and 90d, so that the product provided by the invention is proved to greatly improve the survival rate of the strain in the soil compared with other products.
According to the copy number of the bacillus subtilis gene in each treated tomato soil in the figure 2 and the figure 1, the protective agent can improve the effective viable count of the bacterial strain after the microbial agent is applied to the soil in both crops.
The above-described embodiments are only intended to illustrate the preferred embodiments of the present invention, and not to limit the scope of the present invention, and various modifications and improvements made to the technical solution of the present invention by those skilled in the art without departing from the spirit of the present invention should fall within the protection scope defined by the claims of the present invention.
Claims (7)
1. A method for improving the survival rate of microorganisms in a high inorganic nutrient fertilizer is characterized by comprising the following steps:
(1) mixing the microorganism and a protective agent according to the mass ratio of 1-4:1-5 to prepare a microorganism bacterium agent;
(2) adding the microbial agent into the high inorganic nutrient fertilizer in a proportion of 0.1-10 wt%, and then drying or mixing and granulating to obtain the microbial compound fertilizer.
2. The method according to claim 1, wherein the protective agent comprises the following components in percentage by weight: 1-28% of vegetable oil, 0.5-26% of glycerin, 1-10% of sorbitol, 1-10% of mannitol and the balance of water.
3. The method of claim 1, wherein the high inorganic nutrient fertilizer comprises an inorganic fertilizer, an organic fertilizer, and a carrier.
4. The method as claimed in claim 3, wherein the inorganic fertilizer is 35-45 wt% in the microbial compound fertilizer, the organic fertilizer is 13-24 wt% in the microbial compound fertilizer, and the carrier is 38-42 wt% in the microbial compound fertilizer.
5. The method according to claim 1, wherein the drying condition is cold air drying at 20-55 ℃ for 4-6 h.
6. The method of claim 1, wherein the microorganism comprises bacillus, and the bacillus is mixed with a protective agent in the form of bacillus powder to prepare a microbial agent.
7. A microbial compound fertilizer prepared by the method of any one of claims 1-6.
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| CN117964424A (en) * | 2023-02-20 | 2024-05-03 | 山东省农业科学院 | High-nutrient high-activity chemical fertilizer biological compound fertilizer for water and fertilizer integration |
| WO2024150618A1 (en) * | 2023-01-11 | 2024-07-18 | 合同会社アクト | Multifunctional cosmetic using fermented solution of tuberaceae cells, and whitening serum and polyfunctional slow-release solid chemical fertilizer |
| WO2024174986A1 (en) * | 2023-02-20 | 2024-08-29 | 山东省农业科学院 | Granulation process for chemical fertilizer-biological compound fertilizer, production process, and system |
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