Preparation of benzyl dipeptide and application of benzyl dipeptide in agriculture
By preparing benzyl dipeptide and applying it to agriculture, the environmental pollution problem caused by farmers' excessive application of phosphate fertilizer has been solved, the utilization rate of phosphate fertilizer and crop yield have been improved, and sustainable agricultural development has been achieved.
Patent Information
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- SHANDONG PENGBO BIOTECHNOLOGY CO LTD
- Filing Date
- 2026-01-16
- Publication Date
- 2026-04-28
AI Technical Summary
In order to increase crop yields, farmers overuse phosphate fertilizers, leading to increased production costs and environmental pollution.
Benzyl dipeptide was prepared and applied to agriculture. Its optimal concentration was determined through indoor pot experiments. It was added to soils with different concentrations of phosphate fertilizer to improve phosphorus utilization and crop growth.
It significantly improved the utilization rate of phosphate fertilizer, increased the amount of phosphorus accumulated in crops and the utilization rate of phosphorus, reduced the waste of phosphate fertilizer and environmental pollution, and promoted crop growth and yield.
Smart Images
Figure CN121930176A_ABST
Abstract
Description
Technical Field
[0001] This invention relates to the application technology of benzyl dipeptide in agriculture, specifically to the preparation of benzyl dipeptide and its application in agriculture. Background Technology
[0002] Phosphorus is one of the essential macronutrients for plant growth and development, playing a crucial role in crop yield and quality. However, in natural soils, phosphorus availability is often low, with most phosphorus existing in the form of insoluble compounds that are difficult for plants to directly absorb and utilize.
[0003] In traditional agriculture, farmers often overuse phosphate fertilizers to increase crop yields. This not only increases production costs but also leads to environmental problems such as soil pollution and eutrophication of water bodies. Therefore, developing a new technology that can improve phosphorus utilization and reduce phosphate fertilizer application is of great significance for the sustainable development of agriculture. Summary of the Invention
[0004] The purpose of this invention is to provide the preparation of benzyl dipeptide and its application in agriculture, in order to solve the problem that in the prior art, farmers often apply excessive amounts of phosphate fertilizer in order to increase crop yield, which not only increases production costs, but also leads to environmental problems such as soil pollution and eutrophication of water bodies.
[0005] To achieve the above objectives, the present invention provides the following technical solution: a method for preparing benzyl dipeptide, comprising the following steps:
[0006] Step 1: In a 250 ml single-necked round-bottom flask, add 30 g of accurately weighed L-phenylalanine raw material and about 150 ml of ethylene glycol solvent, ensuring that the liquid level does not exceed two-thirds of the flask's capacity.
[0007] Step 2: Fix the flask on the magnetic stirrer, add the stir bar, and install the spherical condenser tube, and pass cooling water through its top to form a closed loop;
[0008] Step 3: Turn on the stirring device and adjust it to a moderate speed to fully suspend the solid particles. Then turn on the heating module and gradually increase the temperature until the mixture begins to reflux.
[0009] Step 4: Control the heating power to keep the condensed droplets falling steadily, and maintain this mild reflux condition for 10 hours to continue the reaction.
[0010] Step 5: After the reaction is complete, remove the heat source and allow the reaction system to cool down to room temperature naturally. Then place it in an ice-water bath for 30 minutes to promote the crystallization of the product.
[0011] Step 6: Use a Buchner funnel and a vacuum filtration flask for vacuum filtration. First, wet the filter paper with a small amount of ethylene glycol, then wash the filter cake three times with pre-cooled ethylene glycol, each time using about 5 ml. Then wash twice more with ice-cold ethanol or anhydrous ether.
[0012] Step 7: Transfer the washed filter cake to a petri dish, spread it into a uniform thin layer, and place it in a vacuum drying oven. Set the drying temperature to 50-60 degrees Celsius, maintain a stable vacuum in the system, and continue drying for 4-6 hours until constant weight is obtained, yielding a white powdery cyclic crystalline product, benzyl dipeptide.
[0013] The application of benzyl dipeptide in agriculture includes the following steps:
[0014] Step 1: Select wheat, corn, cucumber, and lettuce as experimental materials, and set up a water control and benzyl dipeptide treatment solutions with different concentrations from 5 ng / mL to 1000 ng / mL;
[0015] Step 2: Conduct indoor pot experiments, setting up multiple replicates for each treatment and watering with different concentrations of benzyl dipeptide treatment solution;
[0016] Step 3: Cultivate in an artificial climate chamber with light / dark (16h / 8h), temperature 23℃, humidity 50%-60%, and light intensity 6000-7000 lux. Harvest the plants when there are obvious phenotypic differences.
[0017] Step 4: Measure and record relevant growth indicators and phosphorus content to determine the effective concentration range and optimal concentration of benzyl dipeptide.
[0018] Furthermore, in low-phosphorus soils with a phosphorus fertilizer application rate of 10 mg / kg, adding 20 ng / mL of benzyl dipeptide treatment solution significantly improved the phosphorus accumulation and phosphorus utilization rate of crops.
[0019] Furthermore, in medium-phosphorus soils with an application rate of 20 mg / kg phosphate fertilizer, adding 20 ng / mL of benzyl dipeptide treatment solution promoted crop growth and phosphorus absorption.
[0020] Furthermore, in high-phosphorus soils where the phosphate fertilizer application rate was 30 mg / kg, adding 20 ng / mL of benzyl dipeptide treatment solution further improved the phosphorus accumulation and phosphate fertilizer productivity of crops.
[0021] Furthermore, through indoor pot experiments, wheat was irrigated with a treatment solution containing 20 ng / mL benzyl dipeptide during its growth.
[0022] Furthermore, through indoor pot experiments, corn was irrigated with a treatment solution containing 20 ng / mL benzyl dipeptide during its growth.
[0023] Furthermore, through indoor pot experiments, cucumbers were irrigated with a treatment solution containing 20 ng / mL benzyl dipeptide during their growth.
[0024] Furthermore, through indoor pot experiment, lettuce was irrigated with a treatment solution containing 20 ng / mL benzyl dipeptide during its growth.
[0025] The preparation method of benzyl dipeptide includes the following steps:
[0026] Step 1: Inoculate *Cyclocarya paliurus* onto PDA solid medium and incubate at 26°C for 6 days until the strain is activated. Then, transfer the strain to an Erlenmeyer flask containing 250 ml of PDA liquid seed medium using the agar block method and incubate with shaking for 4-6 days to obtain the first-stage seed culture.
[0027] Step 2: After sterilizing and cooling the 5L fermenter, inoculate it with 7% primary seed liquid and culture for 6 days to obtain the fermentation broth;
[0028] Step 3: Centrifuge the fermentation broth to separate mycelium and filtrate. After washing the mycelium, dry it at 60°C, weigh it and crush it. Extract it three times with an equal volume of ethanol. Mix the extracts three times with a magnetic stirrer, sonicate for 1 hour, vacuum filter, and collect the filtrate, which is the extract.
[0029] Step 4: After the extract is concentrated by rotary evaporation, it is extracted three times with an equal volume of ethyl acetate containing 5% acetone. The supernatant is collected, and the extract is evaporated by rotary evaporation and dried under nitrogen to obtain the crude extract.
[0030] Step 5: The crude extract was initially separated by 200-300 mesh silica gel column chromatography and ODS pressure chromatography, and then combined by TLC, purified by HPLC and analyzed by NMR to obtain benzyl dipeptide;
[0031] The fungus *Scytalidium circinatum* is deposited at the China General Microbiological Culture Collection Center (CGMCC) with accession number CGMCC No. 42391 on November 27, 2025.
[0032] Compared with existing technologies, the preparation of benzyl dipeptide and its application in agriculture provided by this invention have the following beneficial effects:
[0033] The preparation and application of benzyl dipeptide in agriculture has significantly improved phosphate fertilizer utilization. Adding benzyl dipeptide to soils with varying phosphorus levels substantially increased crop phosphorus accumulation and utilization, with the best results observed at a soil application rate of 30 mg / kg phosphate and the addition of 20 ng / mL benzyl dipeptide. This reduces phosphate fertilizer fixation and loss in the soil, allowing more phosphorus to be absorbed and utilized by crops, reducing phosphate fertilizer waste, and alleviating the environmental pressure caused by excessive phosphate fertilizer, thus achieving green and sustainable agricultural development.
[0034] Through systematic experiments, the optimal concentration of benzyl dipeptide was determined and applied to different crops. The results showed significant improvements in crop growth indicators such as plant height and above-ground fresh weight. For example, wheat, corn, cucumber, and lettuce exhibited more robust growth after the addition of benzyl dipeptide, and yield components such as the number of grains per ear and thousand-grain weight also increased, thus enhancing crop yield. Simultaneously, the increased phosphorus content in the crops helps improve crop quality, making them more nutritious and palatable. Therefore, this invention has significant advantages in promoting crop growth and improving yield and quality, bringing greater economic and social benefits to agricultural production. Attached Figure Description
[0035] To more clearly illustrate the technical solutions in the embodiments of this application or the prior art, the drawings used in the embodiments will be briefly introduced below. Obviously, the drawings described below are only some embodiments recorded in this invention. For those skilled in the art, other drawings can be obtained based on these drawings.
[0036] Figure 1 This is a flowchart of the preparation method of the benzyl dipeptide of the present invention. Detailed Implementation
[0037] To enable those skilled in the art to better understand the technical solution of the present invention, the present invention will be further described in detail below with reference to the accompanying drawings.
[0038] Example 1: Preparation of benzyl dipeptide
[0039] Please see Figure 1 As shown, in a 250 mL single-necked round-bottom flask, accurately weigh 30 g of L-phenylalanine and approximately 150 mL of ethylene glycol solvent, ensuring the liquid level does not exceed two-thirds of the flask's capacity. Secure the flask to a magnetic stirrer, add the stir bar, and install a spherical condenser, allowing cooling water to flow through its top to form a closed loop. Turn on the stirrer and adjust the speed to a moderate level to fully suspend the solid particles. Then, activate the heating module and gradually increase the temperature until the mixture begins to reflux. Control the heating power to maintain a steady dripping of the condensate, and continue the reaction under these mild reflux conditions for 10 hours. During this time, the reaction system will gradually change from an initial turbid suspension to a pale yellow turbid liquid.
[0040] After the reaction reaches the predetermined time, remove the heat source and allow the reaction system to cool naturally with continuous stirring. Once the temperature reaches room temperature, the flask can be placed in an ice-water bath and allowed to stand for another 30 minutes to promote maximum crystallization of the product. Assemble the Buchner funnel and vacuum filtration flask, and line it with an appropriate amount of filter paper. First, moisten the filter paper with a small amount of ethylene glycol to ensure a tight seal. Slowly pour the crystalline suspension into the funnel and turn on the vacuum pump for filtration. During operation, carefully control the negative pressure to ensure a smooth flow of the filtrate. After filtration, wash the filter cake three times with pre-cooled ethylene glycol, using approximately 5 ml each time, to thoroughly remove any residual mother liquor. Repeat the washing twice with ice-cold ethanol or anhydrous ether. Carefully transfer the washed filter cake to a watch glass, spread it into a uniform thin layer, and place it in a vacuum drying oven. Set the drying temperature to 50-60 degrees Celsius, maintain a stable vacuum level, and continue drying for 4-6 hours until constant weight. During the drying process, ensure the vacuum circuit remains unobstructed and promptly remove any volatile solvents. The final product is a white powdery cyclic (phenylalanine-phenylalanine) crystal product, which is stored in a desiccator for later use.
[0041] Example 2: Screening of effective concentrations of benzyl dipeptide
[0042] (1) Experimental design
[0043] This experiment was an indoor potted plant experiment. Wheat, corn, cucumber, and lettuce were selected as experimental materials. The experimental treatments are shown in Table 1. A total of 9 treatments were set up, namely water control (CK), and benzyl dipeptide added at concentrations of 5 ng / mL, 10 ng / mL, 20 ng / mL, 50 ng / mL, 100 ng / mL, 200 ng / mL, 500 ng / mL, and 1000 ng / mL. The soil used in the experiment was nutrient soil.
[0044] Wheat and corn: Select plump and uniform seeds, germinate them until they show white sprouts, and then sow them. Take an appropriate amount of soil and put it into a flowerpot. After sowing, cover the seeds with soil. Set up 9 replicates for each treatment. Water each pot with 100 mL of benzyl dipeptide treatment solution at different concentrations. Place the potted plants in an artificial climate room with light / dark (16h / 8h), temperature of 23℃, humidity of 50%-60%, and light intensity of 6000-7000 lux. Harvest the plants when there are obvious phenotypic differences and measure the relevant indicators. Observe the plant growth status every day during the period.
[0045] Cucumbers and lettuce: Select plump and uniform seeds, germinate them until they show white sprouts, and then raise seedlings. Select seedlings with uniform growth and transplant them. Set up 9 replicates for each treatment. After two days of seedling establishment, water each pot with 100 mL of benzyl dipeptide treatment solution at different concentrations. Place the potted plants in an artificial climate room with light / dark (16h / 8h), temperature of 23℃, humidity of 50%-60%, and light intensity of 6000-7000 lux. Harvest the plants when there are obvious phenotypic differences and measure the relevant indicators. Observe the plant growth status every day during the period.
[0046] Table 1 Experimental Design
[0047]
[0048] (2) Test results
[0049] The results are shown in Tables 2, 3, 4, and 5. Benzyl dipeptide can effectively increase phosphorus content in plants, with an effective concentration of 5-500 ng / mL and an optimal concentration of 20 ng / mL (used in subsequent experiments). It is effective on a variety of crops.
[0050] Table 2 Effects of different treatments on wheat growth and phosphorus utilization
[0051]
[0052] Table 3 Effects of different treatments on maize growth and phosphorus utilization
[0053]
[0054] Table 4. Effects of different treatments on cucumber growth and phosphorus utilization
[0055]
[0056] Table 5. Effects of different treatments on lettuce growth and phosphorus utilization.
[0057]
[0058] Example 3: Effects of adding benzyl dipeptide at different phosphorus levels on plant growth and phosphorus uptake and utilization
[0059] 1. Effects of benzyl dipeptide supplementation on wheat growth and phosphorus uptake and utilization at different phosphorus levels
[0060] (1) Experimental design
[0061] This experiment was an indoor pot experiment, using wheat (Jimai 22) as the experimental material, as shown in Table 6. Six treatments were set up, with nine replicates for each treatment. The soil used was field soil from Tai'an City, Shandong Province. Before sowing, phosphate fertilizer was mixed evenly with the field soil according to Table 6. An appropriate amount of soil was placed in each pot, and four wheat seeds were sown in each pot, spreading them evenly on the soil surface without overlap. After sowing, the pots were filled with soil, and then 100 mL of benzyl peptide treatment solution was applied per pot according to the experimental design. The plants were then cultured indoors under an artificial climate with light / dark (16h / 8h), temperature of 23℃, humidity of 50%-60%, and light intensity of 6000-7000 lux. Plants were harvested when phenotypic differences were significant, and relevant indicators were measured. Plant growth was observed daily during the experiment.
[0062] Table 6 Experimental Design
[0063]
[0064] (2) Test results
[0065] Adding benzyl dipeptide at different phosphorus levels can promote wheat growth and phosphorus utilization. When the phosphorus application rate is 30 mg / kg soil and 20 ng / mL benzyl dipeptide is added, the wheat phosphorus accumulation and phosphorus utilization are the highest.
[0066] Table 7. Effects of benzyl dipeptide addition on wheat growth and phosphorus uptake and utilization at different phosphorus levels.
[0067]
[0068] 2. Effects of benzyl dipeptide supplementation on maize growth and phosphorus uptake and utilization at different phosphorus levels
[0069] (1) Test methods
[0070] This experiment was an indoor pot experiment, using maize (Zhengdan 958) as the experimental material, as shown in Table 8. Six treatments were set up, with 12 replicates for each treatment. The soil used was field soil from Tai'an City, Shandong Province. Before sowing the wheat, phosphate fertilizer was mixed evenly with the field soil according to Table 8. An appropriate amount of soil was placed in each pot, and four maize seeds were sown in each pot, spreading them evenly on the soil surface without overlap. After sowing, the pots were filled with soil, and then 100 mL of benzyl peptide treatment solution was applied per pot according to the experimental design. The plants were then cultured indoors under an artificial climate with light / dark (16h / 8h), temperature of 23℃, humidity of 50%-60%, and light intensity of 6000-7000 lux. Plants were harvested when phenotypic differences were significant, and relevant indicators were measured. Plant growth was observed daily during the experiment.
[0071] Table 8 Experimental Design
[0072]
[0073] (2) Test results
[0074] Adding benzyl dipeptide at different phosphorus levels can promote wheat growth and phosphorus utilization. When the phosphorus application rate is 30 mg / kg soil and 20 ng / mL benzyl dipeptide is added, the wheat phosphorus accumulation and phosphorus utilization are the highest.
[0075] Table 9. Effects of benzyl dipeptide addition on maize growth and phosphorus uptake and utilization at different phosphorus levels.
[0076]
[0077] Example 4: Effects of benzyl dipeptide on plant phosphorus uptake and soil phosphorus form transformation
[0078] (1) Experimental design
[0079] This experiment was an outdoor field trial conducted in Tai'an City, Shandong Province. The experimental crop was wheat. As shown in Table 10, three phosphate fertilizer application rates were set up: no phosphate (P0), recommended phosphate application rate (P1, based on Example 3), and farmer's usual phosphate application rate (P2), for a total of 6 treatments, each with 3 replicates, for a total of 18 plots. Each plot area was 36 m2 (3.6 m × 10 m), arranged in a randomized block design. The fertilizer application rates are shown in Table 10. The tested nitrogen fertilizers were ordinary urea (containing 46% N, accounting for 50% of the total nitrogen application) and controlled-release urea (containing 45% N, accounting for 50% of the total nitrogen application); the phosphate fertilizer was diammonium phosphate (containing 18% N and 46% P2O5); and the potassium fertilizer was potassium chloride (containing 60% K2O). All fertilizers were evenly spread on the soil surface once before sowing, followed by rotary tillage (to a depth of about 15-20 cm) to thoroughly mix the fertilizers and samples with the topsoil. After emergence, the seedlings were irrigated with benzyl peptide treatment solution according to the experimental design. Except for the fertilization plan, the sowing density, irrigation, pest and disease control and other agricultural management measures were completely consistent across all treatment plots.
[0080] After the wheat matures, samples are taken. Three duplicate samples are taken from each plot. Soil samples from the 0-20cm soil layer are collected along an "S" shape. After natural air drying, grinding and sieving, the corresponding indicators are measured.
[0081] Table 10 Experimental Design
[0082]
[0083] (2) Test results
[0084] Adding benzyl dipeptide at three different phosphate fertilizer application rates significantly improved the components of wheat yield and the absorption and utilization of phosphorus by plants. It significantly increased the activity of highly active phosphorus phosphate resin-P, directly meaning a substantial increase in "ready-to-eat phosphorus" accessible to plant roots; it also significantly increased phosphatase activity, indicating enhanced soil biotransformation of organic phosphorus. Benzyl dipeptide improves plant utilization of soil phosphorus by altering soil phosphorus forms, thereby promoting phosphorus utilization efficiency.
[0085] Table 11 Effects of benzyl dipeptide on wheat yield components and phosphorus uptake and utilization
[0086]
[0087] Table 12 Effects of benzyl dipeptide on soil phosphorus form transformation
[0088]
[0089] Comparison Example
[0090] Existing technical solutions
[0091] Current technologies primarily improve phosphorus absorption by crops by increasing the application rate of phosphate fertilizers. Specific methods are as follows:
[0092] Phosphate fertilizer selection: Choose common phosphate fertilizers, such as superphosphate and diammonium phosphate.
[0093] Fertilization method:
[0094] Base fertilizer application: Before sowing, spread the phosphate fertilizer evenly on the soil surface, and then mix the phosphate fertilizer into the topsoil through tillage.
[0095] Topdressing: During the crop growth process, phosphate fertilizer is supplemented through irrigation or direct application, depending on the crop's growth status.
[0096] Fertilizer application rate: Determine the recommended application rate of phosphate fertilizer based on soil test results and crop nutrient requirements. For example, for crops such as wheat and corn, the recommended application rate of phosphate fertilizer (P2O5) is generally 80-150 kg / hm².
[0097] Fertilizer management: During fertilization, irrigation management should be combined to ensure that phosphate fertilizer is fully dissolved and absorbed by crops. At the same time, regular soil testing should be conducted to monitor soil phosphorus content and adjust the fertilization plan accordingly. Detailed Implementation
[0098] Example 1: Traditional Phosphate Fertilizer Application Method
[0099] Experimental Design:
[0100] Experimental crops: wheat, corn
[0101] Experimental treatments: A water control (CK) and a conventional phosphate fertilizer treatment (T1) were set up.
[0102] Phosphate fertilizer type: Diammonium phosphate (containing 46% P2O5)
[0103] Fertilizer application rate: Recommended phosphorus application rate is 80 kg / hm² (P1), while farmers are accustomed to applying 150 kg / hm² (P2).
[0104] Experimental soil: Field soil, taken from Tai'an City, Shandong Province.
[0105] Experimental steps:
[0106] Soil preparation: Sift the soil for the experiment to remove impurities, mix it evenly, and then fill it into flower pots with the same amount of soil in each pot.
[0107] Fertilization treatment:
[0108] Base fertilizer application: Before sowing, spread phosphate fertilizer evenly on the soil surface, and then use a rotary tiller to till (to a depth of about 15-20cm) to fully mix the fertilizer and soil.
[0109] Topdressing: During the crop growth process, phosphate fertilizer is supplemented through irrigation according to the crop growth status.
[0110] Sowing and Management: Select plump, uniform seeds for sowing, with 4 seeds per pot. Cover the seeds with soil after sowing. Place the pots in an artificial climate indoor environment with 16h / 8h light / dark conditions, a temperature of 23℃, humidity of 50%-60%, and light intensity of 6000-7000 lux. Observe the crop growth daily and irrigate as needed to keep the soil moist.
[0111] Harvesting and Measurement: Harvest the plants when there are significant differences in crop phenotypes, and measure and record relevant growth indicators (such as plant height, above-ground fresh weight, underground fresh weight, etc.) and phosphorus content.
[0112] Experimental results:
[0113] Traditional methods of applying phosphate fertilizer can improve the phosphorus content and growth indicators of crops to some extent, but the improvement is limited.
[0114] With the increase in the application of phosphate fertilizer, the phosphorus content of crops has increased. However, excessive application of phosphate fertilizer has not significantly increased crop yield. Instead, it has increased production costs and the risk of environmental pollution.
[0115] In summary, while existing technologies that increase the application of phosphate fertilizer to improve crop phosphorus absorption have increased crop phosphorus content to some extent, they also suffer from problems such as low phosphate fertilizer utilization, high production costs, and serious environmental pollution.
[0116] The preparation of benzyl dipeptides from Examples 1 to 4 and their applications in agriculture were compared with those of the control examples, and the results are shown in the table below:
[0117]
[0118] As shown in the table above, Examples 1 to 3, through systematic research and experimentation, demonstrated significant beneficial effects compared to the traditional phosphate fertilizer application method of the control example. Example 1 provides an efficient and stable method for preparing benzyl dipeptide, laying the foundation for subsequent agricultural applications. Example 2, through scientific screening, determined the optimal concentration of benzyl dipeptide, providing a basis for precise agricultural application. Example 3 further verified that in soils with different phosphorus levels, adding benzyl dipeptide can significantly increase the phosphorus accumulation and phosphorus utilization rate of crops, promote crop growth, and reduce phosphate fertilizer waste and environmental pollution risks. In contrast, the traditional phosphate fertilizer application method of the control example suffers from problems such as low phosphate fertilizer utilization rate, high production costs, and serious environmental pollution. Therefore, the technical content of the examples has significant advantages in improving phosphate fertilizer utilization rate, promoting crop growth, and protecting the environment.
[0119] The foregoing has only described certain exemplary embodiments of the present invention by way of illustration. Undoubtedly, those skilled in the art can modify the described embodiments in various ways without departing from the spirit and scope of the present invention. Therefore, the foregoing drawings and descriptions are illustrative in nature and should not be construed as limiting the scope of protection of the claims of the present invention.
Claims
1. A method for preparing benzyl dipeptide, characterized in that, Includes the following steps: Step 1: In a 250 ml single-necked round-bottom flask, add 30 g of accurately weighed L-phenylalanine raw material and 150 ml of ethylene glycol solvent. Step 2: Fix the flask on the magnetic stirrer, add the stir bar, and install the spherical condenser tube, and pass cooling water through its top to form a closed loop; Step 3: Turn on the stirring device and adjust it to a moderate speed to fully suspend the solid particles. Then turn on the heating module and gradually increase the temperature until the mixture begins to reflux. Step 4: Control the heating power to keep the condensed droplets falling steadily, and maintain this mild reflux condition for 10 hours to continue the reaction. Step 5: After the reaction is complete, remove the heat source and allow the reaction system to cool down to room temperature naturally. Then place it in an ice-water bath for 30 minutes to promote the crystallization of the product. Step 6: Use a Buchner funnel and a vacuum filtration flask for vacuum filtration. First, wet the filter paper with a small amount of ethylene glycol, then wash the filter cake three times with pre-cooled ethylene glycol, each time using about 5 ml. Then wash twice more with ice-cold ethanol or anhydrous ether. Step 7: Transfer the washed filter cake to a petri dish, spread it into a uniform thin layer, and place it in a vacuum drying oven. Set the drying temperature to 50-60 degrees Celsius, maintain a stable vacuum in the system, and continue drying for 4-6 hours until constant weight is obtained, yielding a white powdery cyclic crystalline product, benzyl dipeptide.
2. The application of a benzyl dipeptide prepared by the method described in claim 1 in agriculture, characterized in that, Includes the following steps: Step 1: Select wheat, corn, cucumber, and lettuce as experimental materials, and set up a water control and benzyl dipeptide treatment solutions with different concentrations from 5 ng / mL to 1000 ng / mL; Step 2: Conduct indoor pot experiments, setting up multiple replicates for each treatment and watering with different concentrations of benzyl dipeptide treatment solution; Step 3: Cultivate in an artificial climate chamber with light / dark (16h / 8h), temperature 23℃, humidity 50%-60%, and light intensity 6000-7000 lux. Harvest the plants when there are obvious phenotypic differences. Step 4: Measure and record relevant growth indicators and phosphorus content to determine the effective concentration range and optimal concentration of benzyl dipeptide.
3. The application of the benzyl dipeptide in agriculture according to claim 2, characterized in that, In low-phosphorus soils with an application rate of 10 mg / kg phosphate fertilizer, the addition of 20 ng / mL benzyl dipeptide significantly improved the phosphorus accumulation and phosphorus utilization rate of crops.
4. The application of the benzyl dipeptide in agriculture according to claim 2, characterized in that, In medium-phosphorus soils where phosphate fertilizer was applied at a rate of 20 mg / kg, adding 20 ng / mL of benzyl dipeptide solution promoted crop growth and phosphorus absorption.
5. The application of the benzyl dipeptide according to claim 2 in agriculture, characterized in that, In high-phosphorus soils where phosphate fertilizer application rate is 30 mg / kg, adding 20 ng / mL of benzyl dipeptide treatment solution further increases the phosphorus accumulation and phosphate fertilizer productivity of crops.
6. The application of the benzyl dipeptide according to claim 2 in agriculture, characterized in that, In an indoor pot experiment, wheat was irrigated with a treatment solution containing 20 ng / mL benzyl dipeptide during its growth.
7. The application of the benzyl dipeptide according to claim 2 in agriculture, characterized in that, An indoor pot experiment was conducted, in which corn was irrigated with a treatment solution containing 20 ng / mL benzyl dipeptide during its growth.
8. The application of the benzyl dipeptide according to claim 2 in agriculture, characterized in that, An indoor pot experiment was conducted, in which cucumbers were irrigated with a treatment solution containing 20 ng / mL benzyl dipeptide during their growth.
9. The application of the benzyl dipeptide according to claim 2 in agriculture, characterized in that, In an indoor pot experiment, lettuce was irrigated with a treatment solution containing 20 ng / mL benzyl dipeptide during its growth.
10. A method for preparing benzyl dipeptide, characterized in that, Includes the following steps: Step 1: Inoculate *Cyclocarya paliurus* onto PDA solid medium and incubate at 26°C for 6 days until the strain is activated. Then, transfer the strain to an Erlenmeyer flask containing 250 ml of PDA liquid seed medium using the agar block method and incubate with shaking for 4-6 days to obtain the first-stage seed culture. Step 2: After sterilizing and cooling the 5L fermenter, inoculate it with 7% primary seed liquid and culture for 6 days to obtain the fermentation broth; Step 3: Centrifuge the fermentation broth to separate mycelium and filtrate. After washing the mycelium, dry it at 60°C, weigh it and crush it. Extract it three times with an equal volume of ethanol. Mix the extracts three times with a magnetic stirrer, sonicate for 1 hour, vacuum filter, and collect the filtrate, which is the extract. Step 4: After the extract is concentrated by rotary evaporation, it is extracted three times with an equal volume of ethyl acetate containing 5% acetone. The supernatant is collected, and the extract is evaporated by rotary evaporation and dried under nitrogen to obtain the crude extract. Step 5: The crude extract was initially separated by 200-300 mesh silica gel column chromatography and ODS pressure chromatography, and then combined by TLC, purified by HPLC and analyzed by NMR to obtain benzyl dipeptide; The *Cyclocarya paliurus* species described is deposited at the China General Microbiological Culture Collection Center (CGMCC) under accession number CGMCC No. 42391.