Use of berberine, method of use, method of making fertilizer and fertilizer

Abstract

The application discloses a use of berberine as a nitrification inhibitor, a use method of berberine as the nitrification inhibitor, and a fertilizer manufacturing method and a fertilizer formed thereby. Through indoor simulation, field plot and field verification tests, it is found that when the berberine is used in an amount of 10.0-15.0 g per mu, the berberine has a good inhibiting effect on soil ammonia-oxidizing bacteria, can effectively delay the conversion of ammonium nitrogen into nitrate nitrogen, and has an advantage over a traditional nitrification inhibitor in inhibiting a late nitrification stage.

Description

Technical Field

[0001] This invention relates to the field of fertilizers, specifically to the use, application method, fertilizer production method, and fertilizer of berberine as a nitrification inhibitor. Background Technology

[0002] Traditional chemical fertilizers suffer from inherent defects and unscientific or unreasonable application, leading to a series of problems such as low nutrient utilization, serious fertilizer waste, and exacerbated non-point source pollution. Stabilized fertilizers containing nitrification inhibitors can delay the conversion of ammonium nitrogen (including ammonium nitrogen directly contained in nitrogen fertilizers or compound fertilizers, or ammonium nitrogen converted from amide nitrogen) to nitrate nitrogen, allowing nitrogen to exist in the soil as ammonium nitrogen, which is easily adsorbed by soil colloids. Simultaneously, they feature a nutrient release pattern synchronized with crop absorption patterns, significantly improving fertilizer utilization and providing an effective way to solve problems such as fertilizer resource waste and agricultural non-point source pollution. Existing chemically synthesized nitrification inhibitors suffer from drawbacks such as a single inhibitor source, short effective period in soil, and easy decomposition. In recent years, BASF in Germany has launched new inhibitors LIMUS and DMPSA, but their core components are still derivatives of the existing urease inhibitor NBPT and nitrification inhibitor DMPP. The inhibitor source remains relatively singular, mainly chemically synthesized, and still suffers from problems such as susceptibility to environmental factors, easy decomposition upon heating, and rapid degradation in soil, reducing the efficiency of inhibitor use. Therefore, it is necessary to find plant-derived nitrification inhibitors in order to solve the above problems.

[0003] Berberine is an isoquinoline alkaloid isolated from Coptis chinensis, also known as berberine, with the molecular formula C64. 20 H 18 NO4 + It has a relative molecular mass of 336.36 and a density of 1.17 g / cm³. 3Berberine has a melting point of 204-205℃ and a boiling point of 486.8℃, and is soluble in water. It is commonly used as an antibacterial agent to treat gastrointestinal infections and has many other medicinal applications. Berberine is also used in agricultural fertilizers. For example, patent application CN 111675569A discloses a high-efficiency compound foliar fertilizer for fruit trees that promotes flowering and fruiting, and its preparation method. This fertilizer includes chemical fertilizers, berberine hydrochloride, chitosan, polyacrylamide, and other components, which have a positive and effective promoting effect on increasing apple yield and income, and can effectively improve the rate of high-quality apples. Invention patent application CN107759373A discloses a method for preparing a high-fertilizer-utilization-rate slow-release fertilizer for lotus root. The method involves mixing Span and Tween fertilizers evenly, adding olive oil and stirring, then adding egg white, berberine hydrochloride, and other components to form a first material. Fermentation compound bacteria are added to water and mixed evenly, then chicken manure, tea seed meal, and other components are added to form a second material. Ammonium nitrate, ammonium sulfate, and defluorinated phosphate fertilizer are added to the second material and mixed evenly. The mixture is then granulated, sprayed onto the first material for coating, and dried to obtain the high-fertilizer-utilization-rate slow-release fertilizer for lotus root. It is evident that existing technologies all involve using berberine in combination with other components and various fertilizers, without addressing the use of berberine as a specific nitrification inhibitor. Summary of the Invention

[0004] One of the objectives of this invention is to provide the use of berberine as a nitrification inhibitor, that is, berberine can inhibit the conversion of ammonium nitrogen contained in nitrogen fertilizer or compound fertilizer or ammonium nitrogen produced from contained amide nitrogen into nitrate nitrogen, thereby improving the performance of these fertilizers. This new property of berberine is first proposed and utilized in this invention.

[0005] This invention also provides a method for preparing fertilizer, which involves mixing urea or compound fertilizer with berberine, and then applying the mixture directly or after granulation. The weight of berberine is 0.01-0.1% of the weight of urea or compound fertilizer, preferably 0.01-0.05%. The fertilizer prepared by this method has a built-in nitrification inhibition function.

[0006] Preferably, the urea or compound fertilizer is mixed with berberine in one of the following two ways: (a) the urea or compound fertilizer and berberine are directly and mechanically mixed evenly, and then applied directly or after granulation; (b) berberine is dissolved in ethanol to make a solution, and then the solution is evenly sprayed onto the surface of urea granules or compound fertilizer granules and dried.

[0007] More preferably, when berberine is dissolved in ethanol to prepare a solution, the mass ratio of berberine to ethanol is 10.0-15.0 g: 200.0-350.0 mL.

[0008] Another object of the present invention is to provide a fertilizer composed of urea and berberine, or composed of compound fertilizer and berberine, wherein the weight of berberine is 0.01-0.05% of the weight of urea or compound fertilizer. This fertilizer, due to the presence of berberine, has a nitrification inhibition function, which can delay the conversion of ammonium nitrogen to nitrate nitrogen.

[0009] Another objective of this invention is to provide a method for using berberine, wherein 10.0-15.0g of berberine per acre is mixed with fertilizer and applied to crop fields.

[0010] Preferably, the fertilizer is urea or compound fertilizer, the weight of berberine is 0.01-0.05% of the weight of urea or compound fertilizer, and the crop is corn, wheat or rice.

[0011] This invention offers the following beneficial effects: Through screening multiple materials for nitrification inhibition, and analyzing their chemical stability, toxicity, and environmental friendliness, it was discovered that berberine exhibits nitrification inhibition in the fertilizer field, along with good chemical stability, environmental friendliness, and low toxicity. Indoor simulations, field plots, and large-scale field trials demonstrate that berberine at a dosage of 10.0-15.0 g per mu (approximately 0.067 hectares) can inhibit soil ammonia-oxidizing bacteria, effectively delaying the conversion of ammonium nitrogen to nitrate nitrogen. This can increase the content of ammonium nitrogen in the soil, reduce the content of nitrate nitrogen, and improve nitrogen fertilizer utilization. Detailed Implementation

[0012] The present invention will be further described below through specific embodiments:

[0013] 1. Indoor simulation test

[0014] Soil samples from the topsoil layer (0-20cm) were collected from the experimental site in Taiping Village, Changling Town, Songyuan City, Jilin Province. The samples were air-dried and ground through a 2mm sieve. In the experimental group, berberine was added at 0.25% of the weight of urea, and urea at 0.5% of the weight of the air-dried soil. Berberine and urea were first crushed and passed through a 1mm sieve, then mixed thoroughly and finally incorporated into the soil. The mixture was then placed in culture pots, with 200g of soil in each pot. In the control group, urea was added at 0.5% of the weight of the air-dried soil. Urea was first crushed and passed through a 1mm sieve, then the mixture was thoroughly combined with the urea and urea, and then placed in culture pots, with 200g of soil in each pot. Both groups were soaked in water to maintain a soil moisture content of 25%. The soil was placed in a constant temperature environment (30℃), and water was replenished daily based on weight loss. The nitrate nitrogen content in the soil was measured on days 10, 30, 50, and 70. For each sampling and testing, first pour out the soil from the culture pot, mix it thoroughly, take 5.0g of it, place it in an Erlenmeyer flask, and add 2.00mol·L⁻¹. -1100.0 mL of potassium chloride solution was placed in a shaker and extracted for 1 hour. After filtration, the solution was measured using a 3-AA3 flow analyzer. The low nitrate nitrogen content indicates that berberine, as a nitrification inhibitor, delays the conversion of ammonium nitrogen from urea decomposition in the soil into nitrate nitrogen. The experimental results are shown in Table 1. As can be seen from Table 1, berberine showed a good nitrification inhibition effect from day 10 to day 70, effectively reducing the nitrate nitrogen content, allowing nitrogen fertilizer to exist in the soil in the form of ammonium nitrogen, reducing nitrate nitrogen loss, and improving utilization rate.

[0015] Table 1. Monitoring results of nitrate nitrogen content in soil

[0016] Nitrate nitrogen (mg / kg) Day 10 Day 30 Day 50 Day 70 experimental group 159.3 208.4 455.4 602.5 control group 197.1 499.3 703.8 883.7

[0017] 2. Comparative Experiment

[0018] Soil samples from the topsoil layer (0-20cm) were collected at the experimental site in Taiping Village, Changling Town, Songyuan City, Jilin Province. The samples were air-dried and ground through a 2mm sieve. In the experimental group, berberine was added at a urea content of 0.25%, and both urea and berberine were ground through a 1mm sieve. The urea content was 0.5% of the air-dried soil. Berberine and urea were first mixed thoroughly, then mixed evenly with the soil, and the mixture was placed in culture pots, with 200g of soil in each pot. In the control group, DCD (dicyandiamide), a commonly synthesized chemical, was used as a control. DCD was added at a urea content of 0.5%, and both urea and DCD were ground through a 1mm sieve. The urea content was 0.5% of the air-dried soil. DCD and urea were first mixed thoroughly, then mixed evenly with the soil, and the mixture was placed in culture pots, with 200g of soil in each pot. Both the experimental and control groups were soaked in water to maintain a soil moisture content of 25%. The soil was placed in a constant temperature environment (30℃), and water was replenished daily based on weight loss. The nitrate nitrogen content in the soil was measured on days 10, 30, 50, and 70. For each sample measurement, the soil in the culture pot was first poured out, mixed thoroughly, and 5.0 g was placed in an Erlenmeyer flask and 2.00 mol·L⁻¹ was added. -1 100.0 mL of potassium chloride solution was placed in a shaker and extracted for 1 hour. After filtration, the solution was analyzed using a 3-AA3 flow analyzer. The experimental results are shown in Table 2.

[0019] Table 2 Comparison results of berberine and DCD

[0020] Nitrate nitrogen (mg / kg) Day 10 Day 30 Day 50 Day 70 Experimental group - berberine 159.3 208.4 455.4 602.5 Control group - DCD 171.5 240.7 499.4 664.2

[0021] The table above shows that berberine, at half the dosage of DCD, was more effective than DCD from day 10 to day 70, and its toxicity was also better. Berberine has promising application prospects as a nitrification inhibitor.

[0022] 3. Community Trial

[0023] A small-plot verification experiment was conducted in a cornfield in Xujia Village, Laosiping Town, Changtu County, Liaoning Province. Each plot had an area of ​​25m². 2 (5m×5m). The experimental treatments were as follows: conventional fertilization (N), conventional fertilization + berberine, where the amount of berberine was 15.0g per mu.

[0024] Conventional fertilization: Each plot was treated with 977.1g urea, 399.3g superphosphate, and 388.6g potassium sulfate. All three were pulverized and passed through a 1mm sieve, then mixed thoroughly. Conventional fertilization + berberine: Each fertilizer and berberine were pulverized and passed through a 1mm sieve. The amounts and types of nitrogen, phosphorus, and potassium used were the same as conventional fertilization. In addition, 15.0g of berberine was applied per acre, with 0.56g applied per plot. Seeds, pesticides, cultivation, and management methods were consistent between the berberine-treated plots and the control plots without berberine. For conventional fertilization, the plots were mixed with 10 times their weight of dry farmland soil, evenly spread on the surface, and then tilled and sown, with 8 rows per plot and 20 plants per row. For conventional fertilization, after mixing berberine with the fertilizer, another 10 times their weight of dry farmland soil was mixed with the fertilizer, spread on the surface, tilled, harrowed, and then sown. The amounts of nitrate nitrogen and ammonium nitrogen in the soil were measured on days 20, 40, 60, 80, and 120. The results are shown in Tables 3 and 4. The testing method for nitrate nitrogen and ammonium nitrogen in the soil was as follows: Soil samples were taken from the topsoil layer (0-20 cm) using a soil auger. A five-point cross-sampling method was employed, with five augers used for each plot, collecting approximately 300g of soil. After the soil samples were brought back to the laboratory, they were sieved, mixed thoroughly, and 5.0g was taken for analysis using a 2.00 mol·L⁻¹ nitrogen solution. -1 Extracted with potassium chloride solution, and the results were determined using a 3-AA3 flow analyzer.

[0025] Table 3. Monitoring results of nitrate nitrogen in soil

[0026]

[0027] Table 4. Monitoring results of ammonium nitrogen in soil

[0028]

[0029] Yields were measured at harvest. In each plot, starting from the 7th plant in the 4th and 5th rows, five plants were harvested consecutively, for a total of 10 plants. The corn was picked, brought back to the laboratory, threshed, air-dried, and weighed. The results showed: the experimental group had an average yield of 1006.5 kg / mu; the control group had an average yield of 942.7 kg / mu. The average yield increase was 6.7%.

[0030] In summary, berberine demonstrated a good nitrification inhibition effect in maize plot experiments, effectively increasing the ammonium nitrogen content and decreasing the nitrate nitrogen content in the soil, while also effectively increasing maize yield.

[0031] 4. Field trials

[0032] Field trials were conducted to further test the effects of combining berberine with urea or compound fertilizer.

[0033] (1) Shandong maize experiment

[0034] The experimental site was located at the South Campus Experimental Base of Shandong Agricultural University, Taishan District, Tai'an City, Shandong Province. Two treatments were set up: conventional fertilization and conventional fertilization plus berberine. Each treatment had three replicates, and each treatment area was 1 mu (approximately 0.067 hectares). The initial fertility level of each plot was kept consistent. Berberine was applied at a rate of 15.0 g per mu, dissolved in 250.0 mL of ethanol to prepare a solution. 28.5 kg of urea (berberine weight being 0.05% of urea weight), 7.0 kg of potassium chloride (potassium oxide ≥ 62%), and 7.5 kg of diammonium phosphate (18-46-0) were applied per mu. The berberine ethanol solution was evenly sprayed onto the surface of urea granules, dried, and then mixed evenly with potassium chloride and diammonium phosphate. Sowing and fertilization were completed simultaneously using a seed-fertilizer planter. For plots using conventional fertilization, apply 28.5 kg of urea, 7.0 kg of potassium chloride (≥62% potassium oxide), and 7.5 kg of diammonium phosphate (18-46-0) per mu (approximately 0.067 hectares). Mix the urea, potassium chloride, and diammonium phosphate evenly and apply using a seed-fertilizer planter simultaneously. No further topdressing is required, and weeding and pesticide application remain consistent. At harvest, use a combine harvester to harvest each plot individually. After drying and dehydration, weigh the harvested plots to determine the actual yield. The average yield of the three plots is the final data.

[0035] (2) Jiangsu Rice Experiment

[0036] The experimental site was located in Youfang Village, Sheyanghu Town, Baoying County, Yangzhou City, Jiangsu Province. Two treatments were set up: conventional fertilization and conventional fertilization plus berberine, with three replicates per treatment and an area of ​​1 mu (approximately 0.067 hectares) per treatment. Initial fertility levels were kept consistent across all plots. Berberine was applied at a rate of 10.0 g per mu, dissolved in 250.0 mL of ethanol. A conventional compound fertilizer with a nitrogen-phosphorus-potassium ratio of 20-12-16 was used as base fertilizer, applied at a rate of 45.0 kg per mu. The ethanol solution of berberine was evenly sprayed onto the surface of the fertilizer granules and allowed to dry (berberine weight was 0.02% of the compound fertilizer weight). In the control plot, only a conventional compound fertilizer with a nitrogen-phosphorus-potassium ratio of 20-12-16 was applied at a rate of 45.0 kg per mu. The fields were plowed, soaked, and harrowed. Simultaneous side-deep fertilization technology was used during machine transplanting, with transplanting and fertilization occurring concurrently. During the tillering stage, all treatments were treated with urea at a rate of 10.0 kg / mu, and other weeding and pesticide application measures remained consistent. At harvest time, combine harvesters were used to harvest each plot individually. After drying and dehydration, the plots were weighed to obtain the actual yield. The average yield of the three plots was used as the final data.

[0037] (3) Henan wheat experiment

[0038] The experimental site was located in Qiaolaozhuang Village, Shiqiao Town, Ningling County, Shangqiu City, Henan Province. Two treatments were set up: conventional fertilization and conventional fertilization plus berberine, with three replicates per treatment. Each treatment covered an area of ​​1 mu (approximately 0.067 hectares), and the initial fertility level of each plot was kept consistent. Berberine was applied at a rate of 15.0 g per mu, dissolved in 300.0 mL of ethanol. A conventional compound fertilizer with a nitrogen-phosphorus-potassium ratio of 18-22-15 was used as basal fertilizer, applied at a rate of 50.0 kg per mu. The ethanol solution of berberine was evenly sprayed onto the surface of the compound fertilizer granules and allowed to dry (the weight of berberine was 0.03% of the weight of the compound fertilizer). For plots with conventional fertilization, a conventional compound fertilizer with a nitrogen-phosphorus-potassium ratio of 18-22-15 was applied as basal fertilizer at a rate of 50.0 kg per mu. All treatments maintained the same tillage method: plowing, harrowing, and simultaneous sowing and fertilization using a seed-fertilizer co-seeder. All treatments were treated with urea at a rate of 15 kg / mu before spring irrigation, while other weeding and pesticide application measures remained consistent. At harvest time, combine harvesters were used to harvest each plot individually. After drying and dehydration, the harvested plots were weighed to determine the actual yield. The average yield of the three plots was used as the final data, as shown in Table 5.

[0039] Table 5 Yield Measurement Results at Harvest Time

[0040] Yield (kg / mu) Shandong corn Jiangsu rice Henan wheat experimental group 702 617 595 control group 657 578 527 Production increase rate (%) 6.8 6.7 12.9

[0041] In summary, berberine, as a nitrification inhibitor, can effectively inhibit the conversion of ammonium nitrogen to nitrate nitrogen in fertilizers. Applying berberine in combination with urea or compound fertilizer can effectively increase grain yield.

[0042] The above embodiments are illustrative of the present invention and not limiting. Under the concept of the present invention, technical solutions that have not undergone substantial changes are still protected by the present invention.

Claims

1. Uses of berberine as a nitration inhibitor.

2. The use as described in claim 1, characterized in that... Berberine is used to inhibit the conversion of ammonium nitrogen contained in or produced in nitrogen fertilizers or compound fertilizers into nitrate nitrogen.

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