An enzymatic small molecule peptide composition, and a preparation method and application thereof

Abstract

The application discloses an enzymatic small-molecule peptide composition and a preparation method and application thereof, and relates to the technical field of fertilizers.The application comprises the following steps: after plant raw materials are crushed, the plant raw materials are mixed with water and boiled, and then cooled; cellulase is added for enzymolysis, and enzyme is inactivated; liquid is filtered out to obtain protein material A; animal raw materials are soaked in a sodium bicarbonate solution, filtered, soaked in ethanol, filtered, mixed and beaten, water is added, lipase is added for enzymolysis, and enzyme is inactivated to obtain protein material B; the protein material A, the protein material B and wood ash are uniformly mixed, protease and an additive are added for enzymolysis, enzyme is inactivated, and the filtrate is obtained through filtration; the filtrate is freeze-dried, crushed and ground to obtain a soluble small-molecule peptide composition powder.The product contains rich amino acids and small-molecule peptides, can significantly promote crop growth, and has the technical effects of improving benefits and increasing yield compared with conventional fertilizers.

Description

Technical Field

[0001] The present invention relates to the technical field of fertilizers, and in particular to an enzymatically hydrolyzed small molecule peptide composition, a preparation method thereof, and an application thereof. Background Art

[0002] Fertilizer plays a significant role in increasing crop yields and is an indispensable production factor in modern agriculture. Fertilizer plays a vital role in increasing agricultural production and ensuring human food supply.

[0003] Amino acids play a vital ecological role in soil, yet their presence is extremely low. Exogenous amino acid addition can fill the gap in the soil's need for these important organic nitrogen sources. Using enzymatic hydrolysis to hydrolyze waste materials to produce amino acid liquid fertilizer, applied to agricultural soil, not only replenishes amino acids in the soil and improves soil quality, but also provides a new avenue for the development and utilization of waste fish feed and sustainable agricultural development. Amino acids, as the smallest molecules that make up protein, are present in fertilizers and are easily absorbed by crops. They also enhance disease resistance and improve the quality of crops. They supplement essential amino acids for plants, stimulate and regulate rapid plant growth, promote robust plant growth, and facilitate nutrient absorption. They also enhance plant metabolism, improve photosynthesis, promote root development, and accelerate plant growth and reproduction.

[0004] In addition, the small molecule peptides obtained by enzymatic hydrolysis of animal proteins have extremely strong biological activity and diversity, and have a significant effect on improving the quality and yield of agricultural products. For example, Chinese patent application CN201610252537.6 discloses an enzymatically active small molecule peptide organic water-soluble fertilizer and a production method thereof. Using enzymatically active small molecule peptides as a carrier, nitrogen fertilizer, phosphorus fertilizer, potassium fertilizer, humate, and fulvic acid are compounded into the product to form an organic water-soluble fertilizer with strong stability and high effectiveness. The product of the present invention has the efficacy of regulating crop nutritional growth, enhancing crop resistance, increasing yield, improving quality, and can improve the utilization rate of fertilizers and soil nutrients.

[0005] However, on the one hand, the current technology for preparing small molecule peptides using animal and plant raw materials is still immature, and the protein extraction and decomposition process is complex and not conducive to operation. On the other hand, there is little research on the promoting effect of the molecular weight distribution of small molecule peptides on crops. Therefore, further research and development is still needed in the field of preparing small molecule peptide crop fertilizers.

[0006] In view of this, in order to address the deficiencies of the prior art, the present invention provides an enzymatically hydrolyzed small molecule peptide composition, a preparation method thereof, and an application thereof. The small molecule peptide composition is produced from animal and plant raw materials, the process is simple and easy, the product contains rich amino acids and small molecule peptides, and is applied to crop fertilizers to significantly promote crop growth. Compared with conventional fertilizers, it has the technical effect of improving efficiency and increasing yield. Summary of the Invention

[0007] The purpose of the present invention is to provide an enzymatically hydrolyzed small molecule peptide composition and a preparation method and application thereof, which can significantly improve crop quality, increase crop yield and save costs.

[0008] In order to achieve the above-mentioned purpose of the invention, the technical solution of the present invention is as follows:

[0009] In one aspect, the present invention provides a method for preparing an enzymatically hydrolyzed small molecule peptide composition, comprising the following steps:

[0010] (1) After the plant raw materials are crushed, water is added, mixed and boiled, cooled, cellulase is added for enzymolysis, the enzyme is inactivated, and the liquid is filtered to obtain protein material A; the plant raw materials are wheat, sorghum, soybean and sunflower head;

[0011] (2) soaking the animal raw material in sodium bicarbonate solution, filtering, soaking in ethanol, filtering, mixing and beating, adding water, adding lipase for enzymolysis, and inactivating the enzyme to obtain protein material B; the animal raw material is shrimp shells and fish viscera;

[0012] (3) Protein material A, protein material B, and wood ash are mixed evenly, and protease and auxiliary agents are added for enzymolysis, the enzyme is inactivated, the filtrate is filtered, freeze-dried, and crushed to obtain a soluble small molecule peptide composition powder.

[0013] Wherein, the auxiliary agent is at least one of lignin sulfonate and humate.

[0014] In the present invention, wheat, sorghum and soybean are all sun-dried seeds.

[0015] Preferably, in step (1), the mass ratio of the wheat, sorghum, soybean and sunflower disc is 1-10:1-10:1-10:1, and more preferably 2:2:2:1.

[0016] Preferably, in step (1), the boiling time is 20-60 min, more preferably 25-35 min.

[0017] Preferably, in step (1), the conditions for the enzymatic hydrolysis of the cellulase are: solid content of 1-5%, enzymatic hydrolysis time of 30-120 min, enzyme dosage of 0.1-3%, and enzymatic hydrolysis temperature of 40-60°C. Most preferably, the conditions for the enzymatic hydrolysis of the cellulase are: solid content of 3%, enzymatic hydrolysis time of 90 min, enzyme dosage of 0.5%, and enzymatic hydrolysis temperature of 50°C.

[0018] Preferably, in steps (1)-(3), the enzymatic hydrolysis is carried out under stirring.

[0019] Preferably, in step (2), the solid-to-liquid ratio (weight ratio) of the sodium bicarbonate solution is 1:5-20, more preferably 1:10; the soaking time is 2-8 hours, more preferably 3-4 hours.

[0020] Preferably, in step (2), the temperature of ethanol immersion is 40-80°C, more preferably 50°C; the solid-liquid ratio (weight ratio) of ethanol immersion is 1:5-20, more preferably 1:10; the time of ethanol immersion is 5-24h, more preferably 8-10h.

[0021] Preferably, in step (2), the conditions for lipase hydrolysis are: solid content 3-10%, hydrolysis time 30-60 min, hydrolysis temperature 25-40° C., and enzyme dosage 0.1-5%. Most preferably, the conditions for lipase hydrolysis are: solid content 6%, hydrolysis time 50 min, hydrolysis temperature 30° C., and enzyme dosage 0.3%.

[0022] Preferably, the weight ratio of the protein material A to the protein material B is 1:1-3, more preferably 1:1.5. Preferably, in step (3), the protease is alkaline protease and papain, more preferably, the weight ratio of the alkaline protease to papain is 3:1.

[0023] Preferably, in step (3), the conditions for protease hydrolysis are: hydrolysis time 30-120 min, hydrolysis temperature 40-60° C., and enzyme dosage 0.1-5%. Most preferably, the conditions for lipase hydrolysis are: hydrolysis time 60 min, hydrolysis temperature 50° C., and enzyme dosage 1%.

[0024] Preferably, in step (3), the amount of wood ash added is 1-2.5% of the total weight of protein material A and protein material B, more preferably 2%.

[0025] Preferably, in step (3), the amount of the additive added is 0.1-10%, more preferably 3%.

[0026] Preferably, in step (3), the auxiliary agents are lignin sulfonate and humate, and the mass ratio of the two is 1:1-10, more preferably 1:5.

[0027] In another aspect, the present invention provides an enzymatically hydrolyzed small molecule peptide composition prepared according to the above preparation method, characterized in that it comprises amino acids and small molecule peptides.

[0028] Preferably, the content of the amino acid is 10-15%.

[0029] Preferably, the molecular weight distribution of the small molecule peptide is: below 430, 55-65%; greater than 430 and less than 1700, 29-32%; 1700-5800, no more than 10%.

[0030] Finally, the present invention provides the use of the enzymatically hydrolyzed small molecule peptide composition in the preparation of crop fertilizers.

[0031] Preferably, the crops include cotton, corn, potato, sweet potato, sunflower, wheat and rice.

[0032] The beneficial effects of the present invention are:

[0033] (1) The present invention provides a method for preparing an enzymatically hydrolyzed small molecule peptide composition. The process is simple and easy. Plant protein and animal protein are processed separately. The obtained product contains rich amino acids and small molecule peptides. When applied to crop fertilizers, it can significantly promote crop growth. Compared with conventional fertilizers, it has the technical effect of improving efficiency and increasing yield.

[0034] (2) Processing animal raw materials and plant raw materials separately, and then adding a certain amount of wood ash and protein materials for enzymatic hydrolysis together can improve the enzymatic hydrolysis efficiency of protease, save the amount of protease used, and improve the enzymatic hydrolysis effect.

[0035] (3) The combined enzymatic hydrolysis of alkaline protease and papain improved the enzymatic hydrolysis effect compared with single enzymatic hydrolysis, significantly increased the amino acid content, and improved the distribution of small molecule peptides.

[0036] (4) The present invention uses lignin sulfonate and humate as enzymatic hydrolysis aids, which, in addition to adjusting the pH, are also beneficial to increasing the amino acid content. BRIEF DESCRIPTION OF THE DRAWINGS

[0037] Figure 1 It is a process flow chart of the present invention.

[0038] Figure 2 This is a diagram showing the effect of the composition of the present invention on the development of corn seeds.

[0039] Figure 3 This is a diagram showing the effect of the composition of the present invention on the development of the root system of cotton seedlings.

[0040] Figure 4 This is a graph showing the effects of different fertilization treatments on the number of true leaves, fruiting branches, buds and bolls, and plant height at the cotton seedling stage.

[0041] Figure 5 This is a diagram showing the effects of different fertilization treatments on agronomic traits of cotton at the boll stage (Korla).

[0042] Figure 6 This figure shows the effect of different fertilization on cotton dry matter accumulation.

[0043] Figure 7 This figure shows the effects of different fertilization treatments on the photosynthetic performance of cotton. DETAILED DESCRIPTION

[0044] The following non-limiting examples are provided to enable those skilled in the art to more fully understand the present invention, but are not intended to limit the present invention in any way. The following is merely an illustrative description of the scope of protection claimed in this application. Those skilled in the art may make various changes and modifications to the invention of this application based on the disclosed content, and such changes and modifications should also fall within the scope of protection claimed in this application.

[0045] The present invention will be further described below in the form of specific embodiments. The various chemical reagents used in the embodiments of the present invention are all obtained through conventional commercial channels unless otherwise specified. Unless otherwise specified, the content (percentage) described hereinafter is all mass content (mass percentage). Unless otherwise specified, it is understood that it is carried out at room temperature. Hereinafter, the term "solid content" refers to the mass percentage of the solid part in the whole mixture, and the amount of enzyme added hereinafter is the mass percentage of the weight of the enzyme in the raw material to be enzymolyzed, such as "0.3% lipase" represents that the mass of lipase accounts for 0.3% of the total mass of the plant raw material, and "1% composite protease" represents that the mass of the composite protease accounts for 0.3% of the total mass of protein material A and protein material B. The amount of auxiliary agent added is calculated in the same way as the amount of enzyme added.

[0046] In the embodiments of the present invention, the cellulase used is Xiasheng cellulase FFG-0665; the lipase used is Xiasheng lipase SDG-2426, the alkaline protease used is Xiasheng alkaline protease FDG-2202, and the papain used is Xiasheng papain FDG-2203, which does not limit the scope of protection of the present invention.

[0047] Example 1

[0048] (1) Wheat, sorghum, soybeans, and sunflower heads were crushed and mixed in a mass ratio of 2:2:2:1. Water was added to adjust the solid content to 3%. The mixture was mixed thoroughly, boiled, and cooled after 30 minutes. The mixture was cooled to 50° C. and maintained at a constant temperature. 0.5% cellulase was added while stirring and enzymatically hydrolyzed for 90 minutes. The temperature was raised to inactivate the enzyme, and the liquid was filtered to obtain protein material A.

[0049] (2) Shrimp shells and fish viscera were mixed in a mass ratio of 1:1, and sodium bicarbonate solution was added and soaked at room temperature for 3 hours, with a solid-liquid ratio (weight ratio) of 1:10. After soaking, the mixture was filtered and washed with water three times. An equal weight of 75% ethanol was added again, and the mixture was soaked at 50°C for 10 hours. The mixture was filtered, mixed and beaten, and water was added to adjust the solid content to 6%. The temperature was maintained at 30°C, and 0.3% lipase was added for enzymatic hydrolysis for 50 minutes. The enzyme was inactivated by heating to obtain protein material B.

[0050] (3) Protein material A and protein material B are mixed in a weight ratio of 1:1.5, 2% wood ash is added and mixed evenly, the temperature is maintained at 50°C, 1% composite protease (0.75% alkaline protease + 0.25% papain), 0.5% potassium lignin sulfonate, and 2.5% potassium humate are added, stirred evenly and enzymatically hydrolyzed for 60 minutes under stirring, the enzyme is inactivated by heating, filtered, the filtrate is collected, freeze-dried, and crushed to obtain a soluble small molecule peptide composition powder.

[0051] Example 2

[0052] (1) Wheat, sorghum, soybeans, and sunflower heads were crushed and mixed in a mass ratio of 2:2:2:1. Water was added to adjust the solid content to 5%. The mixture was mixed thoroughly, boiled, and cooled after 30 minutes. The mixture was cooled to 50° C. and maintained at a constant temperature. 0.5% cellulase was added while stirring and enzymatically hydrolyzed for 70 minutes. The temperature was raised to inactivate the enzyme, and the liquid was filtered to obtain protein material A.

[0053] (2) Shrimp shells and fish viscera were mixed in a mass ratio of 1:1, and sodium bicarbonate solution was added and soaked at room temperature for 4 hours, with a solid-liquid ratio (weight ratio) of 1:15. After soaking, the mixture was filtered and washed with water three times. An equal weight of 75% ethanol was added again, and the mixture was soaked at 50°C for 8 hours. The mixture was filtered, mixed and beaten, and water was added to adjust the solid content to 3%. The temperature was maintained at 30°C, and 1% lipase was added for enzymatic hydrolysis for 50 minutes. The enzyme was inactivated by heating to obtain protein material B.

[0054] (3) Protein material A and protein material B are mixed in a weight ratio of 1:3, 1% wood ash is added and mixed evenly, the temperature is maintained at 50°C, 1% composite protease (0.75% alkaline protease + 0.25% papain), 1.0% potassium lignin sulfonate, and 2.0% potassium humate are added, stirred evenly and enzymatically hydrolyzed for 80 minutes under stirring, the enzyme is inactivated by heating, filtered, the filtrate is collected, freeze-dried, and crushed to obtain a soluble small molecule peptide composition powder.

[0055] The amino acid content was tested according to NY 1429-2010, and the results are as follows:

[0056]

[0057]

[0058] It can be seen that the free amino acid content in the compositions prepared in Example 1 and Example 2 is both above 10%.

[0059] The molecular weight distribution of peptides was determined by liquid chromatography according to Appendix A of GB / T 22492-2008. Standards included cytochrome C, insulin, bacitracin, tetrapeptides glycine-glycine-tyrosine-arginine, and glycine-glycine-glycine. The column was TSKgelG2000SWXL (7.8 mm × 30 cm). The mobile phase consisted of acetonitrile:water:trifluoroacetic acid (20:80:0.1) at a flow rate of 0.5 ml / min, a column temperature of 35°C, and an injection volume of 10 μl.

[0060] The calculation formula is:

[0061] X=(A i / A 总 )×100%

[0062] X: relative percentage of each molecular weight segment

[0063] Ai: area of ​​each molecular weight segment

[0064] A 总 : Total area of ​​total molecular weight segment

[0065] The results of Example 1 are as follows:

[0066]

[0067] The results of Example 2 are as follows:

[0068]

[0069]

[0070] Comparative Example 1

[0071] (1) Wheat, sorghum, soybeans, and sunflower heads were crushed and mixed in a mass ratio of 2:2:2:1. Water was added to adjust the solid content to 3%. The mixture was mixed thoroughly, boiled, cooled after 30 minutes, filtered, and the liquid was removed to obtain a mixture A.

[0072] (2) Shrimp shells and fish viscera were mixed in a mass ratio of 1:1, and soaked in sodium bicarbonate solution at room temperature for 3 h with a solid-to-liquid ratio (weight ratio) of 1:10. After soaking, the mixture was filtered and washed with water three times. An equal weight of 75% ethanol was added again, and the mixture was soaked at 50° C. for 10 h, filtered, mixed, and beaten to obtain a mixture B.

[0073] (3) Mix mixture A and mixture B in a weight ratio of 1:1.5, add 2% wood ash and mix evenly, maintain the temperature at 50°C, add 1% composite protease (0.75% alkaline protease + 0.25% papain), 0.5% potassium lignin sulfonate, and 2.5% potassium humate, stir evenly and enzymatically hydrolyze for 60 minutes under stirring, heat to inactivate the enzyme, filter, collect the filtrate, freeze-dry, and crush to obtain a soluble small molecule peptide composition powder.

[0074] Comparative Example 2

[0075] The difference from Example 1 is that only 1% alkaline protease is added during the enzymatic hydrolysis in step (3), and the conditions of the other steps are the same.

[0076] Comparative Example 3

[0077] The difference from Example 1 is that in step (3), potassium lignin sulfonate is not added as the auxiliary agent, but only 2.5% potassium humate is added, and the conditions of the other steps are the same.

[0078] Comparative Example 4

[0079] The difference from Example 1 is that wood ash is not added in step (3), and the conditions of the other steps are the same.

[0080] The amino acid content and peptide molecular weight distribution were determined using the same methods as before. The results are as follows:

[0081]

[0082]

[0083] As can be seen from the table above, the amino acid content of Comparative Examples 1-4 did not reach 10%.

[0084] The molecular weight distribution of the peptide of Comparative Example 1 is as follows:

[0085]

[0086] The molecular weight distribution of the peptide of Comparative Example 2 is as follows:

[0087]

[0088] The molecular weight distribution of the peptide of Comparative Example 3 is as follows:

[0089]

[0090] The molecular weight distribution of the peptide of Comparative Example 4 is as follows:

[0091]

[0092] Experimental Example 1

[0093] The root development of corn seeds and cotton seedlings was tested and the results are shown in the attached figure. Figure 2 、 3 Test group: corn seeds were soaked in a 0.1% aqueous solution of the composition of Example 1 of the present invention; cotton seedlings were drip-applied with the composition of Example 1;

[0094] Control group: corn seeds were soaked in water only; cotton seedlings were not subjected to the composition solution, and other culture conditions were the same as those of the experimental group.

[0095] From the attached Figure 2 、 3 It can be seen that, compared with the control group, the composition of the present invention can significantly increase the germination rate of corn seeds and promote the development of the root system of cotton seedlings.

[0096] Experimental Example 2

[0097] 2.1 Test location:

[0098] Huyanghe Experimental Station of the Cotton Research Institute of the Chinese Academy of Agricultural Sciences (44°44′N, 84°48′E, temperate continental arid climate, average annual precipitation of 182.1 mm, medium soil fertility, clay loam) and Korla Experimental Station (41°10′N, 85°14′E, temperate continental arid climate, average annual precipitation of 58.6 mm, medium soil fertility, slightly saline-alkali, clay loam).

[0099] Experimental varieties: China Cotton Research Institute 109 and China Cotton Research Institute 99, the fiber quality of both is above "double 30".

[0100] 2.2 Test method:

[0101] (1) Huyanghe Experimental Station

[0102] Randomized block design, two treatments were set up in the fertilizer test, namely CK control (conventional field fertilization) and T1 treatment (using the composition of Example 1 of the present invention to replace urea), adopting the "wide, early and excellent" mode (1 film, 3 rows, equal row spacing), and the planting density was 12,000 plants / mu. The fertilization measures of the CK treatment were carried out in accordance with the experimental field management. The nitrogen fertilizers were urea and monoammonium, the phosphate fertilizer was monoammonium phosphate, and the potash fertilizer was potassium sulfate; the T1 treatment did not apply urea, but used the composition of Example 1 instead, and drip-fertilized according to the fertilization plan. Other field management measures such as weeding, pest control, topping, etc. were carried out in accordance with the management of the experimental station. Each treatment was repeated 3 times, with a total of 6 plots, and the area of ​​each plot was 57m×6.5m=370.5m 2 , with a total area of ​​3.3 acres, and sowing was carried out on April 24, 2020.

[0103] (2) Korla Experimental Station

[0104] A large-scale demonstration comparison was conducted, and 67 mu of test land was selected. Two water outlets (independent irrigation systems) were used for fertilization dripping. 37 mu of the land was used for field treatment (CK), and 30 mu of land was treated with the composition of Example 1 instead of urea (T1). The fertilization plan was followed, and other field management measures such as weeding, pest control, and topping were carried out in accordance with the management of the experimental station.

[0105] Fertilization plan:

[0106] Huyanghe CK group: 42 kg / mu of urea + 26 kg / mu of monoammonium phosphate + 3 kg / mu of potassium dihydrogen phosphate + 20 kg / mu of potassium sulfate + 20 kg / mu of monoammonium phosphate + 1 kg / mu of potassium fulvic acid + 800 g / mu of boron zinc fertilizer;

[0107] Korla CK group: urea 42 kg / mu + monoammonium 20 kg / mu + potassium dihydrogen phosphate 3 kg / mu + potassium sulfate 20 kg / mu + monoammonium phosphate 14.5 kg / mu + potassium fulvic acid 2 kg / mu + boron zinc fertilizer 800 g / mu + organic fertilizer 9.3 kg / mu;

[0108] Group T1: 410 g / mu of the composition of Example 1 + 2 kg / mu of potassium dihydrogen phosphate + 15 kg / mu of monoammonium phosphate + 12 kg / mu of potassium sulfate + 1 kg / mu of potassium fulvic acid + 800 g / mu of boron zinc fertilizer.

[0109] 2.3 Survey indicators:

[0110] (1) Records of cotton disease resistance and cultivation practices

[0111] Record in detail the field management such as fertilization time and amount, dripping time and amount of water, spraying time and dosage, and calculate the output-input ratio.

[0112] (2) Investigation of cotton agronomic traits

[0113] During the cotton growing season, surveys were conducted six times (seedling stage, bud stage, initial flowering stage, peak flowering stage, boll stage, and boll opening stage) on 10 established cotton plants in each plot. Records primarily included plant height, number of fruiting branches, and the number of buds, flowers, bolls, rotten bolls, and boll opening on each branch.

[0114] (3) Determination of cotton photosynthetic performance and light energy utilization

[0115] During the cotton growth period, photosynthetic efficiency and light energy utilization rate of different treatments were measured using photosynthetic meter Li-6800 and light meter Li-300.

[0116] (4) Cotton yield, composition, and fiber quality

[0117] The theoretical and actual yields were measured during the boll opening period, and 6.67m 2Yield was measured in the plots three times, and fiber quality was also tested. The economic benefits of seed cotton in the T1 group were compared.

[0118] 2.4 Results

[0119] (1) Effects of different fertilization treatments on cotton agronomic morphology

[0120] Dynamic changes of cotton agronomic traits under different fertilization treatments Figure 4 、 Figure 5 As shown, at the Huyanghe Experimental Station, plant height differences between the two treatments were not significant during the early growth phase. However, as growth progressed, the T1 treatment significantly promoted plant height growth. At 60 days (bud stage) and 90 days (boll stage) after emergence, plant height in the T1 treatment was 10% and 12% higher than in the CK treatment. From 20 to 90 days after emergence, plant height in the CK and T1 treatments increased by 62 cm and 72 cm, respectively, indicating that T1 promoted vegetative growth in cotton. The number of true leaves in the seedling stage was higher in the T1 treatment than in the CK treatment. 70 days after sowing, the number of fruiting branches in the T1 treatment was 22.2% higher than in the CK treatment, and 3.8% higher. At 101 days after sowing (peak boll stage), the number of bolls per plant in the T1 treatment was 2.3 higher than in the CK treatment, and the number of fruiting branches was 1.8 higher. Similarly, at the Korla Experimental Station, T1 > CK in all of these patterns. The results from the two sites demonstrate that the composition of the present invention effectively promotes cotton growth and development.

[0121] (2) Effects of different fertilization treatments on cotton dry matter weight

[0122] The accumulation pattern of cotton dry matter under different fertilization treatments is as follows Figure 6 As shown, early dry matter and nitrogen distribution centers on the vegetative organs. After the flowering and boll formation stage, dry matter and nitrogen distribution gradually shift to the reproductive organs, ultimately resulting in a higher proportion of dry matter in the reproductive organs. During the cotton seedling stage, the highest proportion of photosynthate is allocated to the stem. From bud formation to initial flowering, only 6-9% is allocated to buds and bolls. This percentage gradually increases in the later stages, and after the boll opening stage, the vast majority of dry matter (61-63%) is allocated to bolls. Throughout the growth period, the weight of leaves, stems, reproductive organs, and total dry matter in the T1 treatment is greater than that in the CK treatment. In particular, at the full boll formation stage (August 9), the weight of reproductive organs and total dry matter in the T1 treatment were 43.5% and 54.3% higher than those in the CK treatment, indicating that application of the composition of the present invention effectively increases the proportion of photosynthate converted to reproductive organs and accelerates the buildup of dry matter in cotton.

[0123] (3) Effects of different fertilization treatments on cotton photosynthetic performance

[0124] The photosynthetic performance of cotton under different fertilization treatments was analyzed ( Figure 7) Results showed that the T1 treatment significantly affected the net photosynthetic rate, transpiration rate, intercellular carbon dioxide concentration, and stomatal conductance of cotton at all growth stages, all of which were higher than those of the CK treatment. In particular, at the peak flowering stage, the net photosynthetic rate, transpiration rate, intercellular carbon dioxide concentration, and stomatal conductance of the T1 treatment were 3.5%, 13.9%, 5.6%, and 21.6% higher than those of the CK treatment, respectively.

[0125] (4) Effects of different fertilization treatments on cotton light energy utilization efficiency

[0126] The results are as follows:

[0127]

[0128] As shown in the table above, the light absorption rate under the T1 treatment was significantly higher than that under the CK treatment, while the transmittance was significantly lower than that under the CK treatment. This indicates that the T1 treatment can form a reasonable canopy structure, promote spiders' absorption and utilization of light energy, and enhance the photosynthetic efficiency of the cotton colony.

[0129] (5) Effects of different fertilization treatments on cotton yield and fiber quality

[0130] Impact on production:

[0131]

[0132] Impact on fiber quality:

[0133]

[0134] As shown in the table above, there was no significant difference in fiber quality between the two treatments, indicating that the composition of the present invention did not affect fiber quality. Compared with the CK, the T1 treatment significantly increased the number of bolls per plant. The Huyanghe T1 treatment increased yield by 5.6% compared to the CK, and the Korla T1 treatment increased yield by 4.7%. Based on a seed cotton purchase price of 6.5 yuan / kg, this translates to an increase of approximately 130 yuan per mu.

[0135] (6) Effects of different fertilization treatments on economic benefits

[0136] Through statistics of fertilizer input and output value, the output-input ratio was calculated. The output-input ratio of the Populus euphratica CK group was 10.5, that of the T1 group was 12.5, that of the Korla CK group was 15, and that of the T1 group was 16.2. By using the composition of the present invention, the fertilizer input cost was reduced by 6-22 yuan / mu, and the net income was increased by about 139 yuan / mu.

[0137] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions, improvements, etc. made within the spirit and principles of the present invention should be included in the scope of protection of the present invention.

Claims

1. A method for preparing an enzymatically hydrolyzed small molecule peptide composition, characterized in that: The following steps are involved: (1) After the plant raw materials are crushed, water is added to the mixture and the mixture is boiled, cooled, cellulase is added for enzymolysis, the enzyme is inactivated, and the liquid is filtered to obtain protein material A; the plant raw materials are wheat, sorghum, soybean and sunflower heads; (2) Soaking the animal raw material in sodium bicarbonate solution, filtering, soaking in ethanol, filtering, mixing and beating, adding water, adding lipase for enzymolysis, and inactivating the enzyme to obtain protein material B; the animal raw material is shrimp shells and fish viscera; (3) Mix protein material A, protein material B and wood ash evenly, add protease and auxiliary agents for enzymatic hydrolysis, inactivate the enzyme, filter the filtrate, freeze-dry, and crush to obtain a soluble small molecule peptide composition powder. in, In step (1), the conditions for the enzymatic hydrolysis of the cellulase are: solid content of 1-5%, enzymatic hydrolysis time of 30-120 min, enzyme dosage of 0.1-3%, and enzymatic hydrolysis temperature of 40-60°C; in step (2), the conditions for the enzymatic hydrolysis of the lipase are: solid content of 3-10%, enzymatic hydrolysis time of 30-60 min, enzymatic hydrolysis temperature of 25-40°C, and enzyme dosage of 0.1-5%; in step (3), the conditions for the enzymatic hydrolysis of the protease are: enzymatic hydrolysis time of 30-120 min, enzymatic hydrolysis temperature of 40-60°C, and enzyme dosage of 0.1-5%; In step (3), the weight ratio of the protein material A to the protein material B is 1:1-3, the protease is alkaline protease and papain, and the weight ratio of alkaline protease to papain is 3:1; The amount of the plant ash added is 1-2.5% of the total weight of the protein material A and the protein material B, the amount of the additive added is 0.1-10%, and the additives are lignin sulfonate and humate, and the mass ratio of the two is 1:1-10.

2. The preparation method according to claim 1, characterized in that In step (1), the mass ratio of the wheat, sorghum, soybean and sunflower disc is 1-10:1-10:1-10:

1.

3. The preparation method according to claim 1, characterized in that In step (1), the boiling time is 20-60 minutes.

4. The preparation method according to claim 1, characterized in that In step (2), the solid-liquid ratio of the sodium bicarbonate solution is 1:5-20, the soaking time is 2-8 hours, the temperature of the ethanol soaking is 40-80°C, the solid-liquid ratio of the ethanol soaking is 1:5-20, and the ethanol soaking time is 5-24 hours.

5. The enzymatically hydrolyzed small molecule peptide composition prepared by the preparation method according to any one of claims 1 to 4, characterized in that: Including amino acids and small molecule peptides.

6. The enzymatically hydrolyzed small molecule peptide composition according to claim 5, characterized in that: The amino acid content is 10-15%, and the molecular weight distribution of the small molecule peptide is: below 430, 55-65%; greater than 430 and less than 1700, 29-32%; 1700-5800, no more than 10%.

7. Use of the enzymatically hydrolyzed small molecule peptide composition according to claim 5 or 6 in the preparation of crop fertilizer.

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