Nano enzyme, preparation method thereof, agricultural preparation and application of agricultural preparation

Abstract

The invention relates to a nano-enzyme, a preparation method thereof, an agricultural preparation and application of the agricultural preparation. The catalyst comprises the following preparation raw materials in parts by weight: 1-6 parts of metal salt; 0.5 to 4 parts of a surfactant; 1 part to 1.04 parts of an acidic reagent; the metal salt comprises one or more of ferric salt, manganese salt, calcium salt, zinc salt and copper salt; the metal organic framework material of the nano enzyme is Prussian blue. The preparation raw materials of the nano-enzyme comprise specific components with a specific proportioning relation, metal salt provides an active center and a structural basis of the nano-enzyme, that is, existence of metal ions can provide the active center and endow the nano-enzyme with catalytic activity, and the nano-enzyme is similar to metal ions in a natural enzyme to play a catalytic role at active sites; the surfactant controls the size and dispersity of the nano-enzyme, the acidic reagent adjusts the reaction conditions, and the three components cooperate with each other to provide guarantee for successful preparation and performance optimization of the nano-enzyme.

Description

Technical Field

[0001] The present application relates to the technical field of nanomaterials, and in particular to a nanozyme and a preparation method thereof, an agricultural preparation and an application thereof. Background Art

[0002] Nanozymes are environmentally friendly catalytic materials with nano-sized sizes and similar enzymatic reaction kinetics and specificity to natural enzymes. They are widely used in biology, medicine and other fields. In the agricultural field, nanozymes have strong environmental tolerance and micro-fertilizer effect, making them particularly suitable for large-scale industrial production. However, although nanozymes have shown broad application prospects in the agricultural field, the integrated fertilizer and drug technology based on nanozymes has not yet been widely used.

[0003] Seed coating agents are widely used in agricultural production. They provide necessary nutrients by coating seeds and effectively prevent and control pests and diseases, which plays a vital role in promoting the healthy growth of crops and increasing crop yields. However, traditional seed coating agents usually contain high concentrations of pesticides, which can easily cause pesticide damage to seeds, affecting their germination rate and seedling growth. In addition, traditional seed coating agents have a short effective period, low pesticide utilization, and may introduce toxic warning color dyes, which is not only detrimental to the ecological environment, but also difficult to meet the needs of green and sustainable agricultural development.

[0004] Therefore, there is an urgent need for products and technologies that can solve the problems of pesticide damage and abiotic stress caused by traditional seed coating agents, improve the effective utilization rate of pesticides, and promote the healthy growth of seeds. Summary of the invention

[0005] Based on this, the present application provides a nanozyme, which can effectively degrade free radicals produced by pesticide stress and reduce their toxic effects on biological systems. When used as seed coating materials, it can not only reduce the risk of phytotoxicity of traditional pesticides to seeds, but also significantly improve the germination rate of seeds and the growth quality of seedlings; and accordingly, a nanozyme and its preparation method, agricultural preparation and its application are provided.

[0006] The specific technical solutions are as follows: The first aspect of the present application provides a nanozyme, comprising the following preparation raw materials by weight: 1 to 6 parts of metal salt; 0.5 to 4 parts of surfactant; Acidic reagent 1 part to 1.04 parts; The metal salt includes one or more of iron salt, manganese salt, calcium salt, zinc salt and copper salt; the metal organic framework material of the nanozyme is Prussian blue.

[0007] In some embodiments, the surfactant includes tea seed cake extract; The acidic agent includes citric acid.

[0008] On the other hand, the present application also provides a method for preparing a nanozyme, comprising the following steps: The above-mentioned raw materials for preparing nanozymes are mixed, reacted and dried to prepare nanozymes.

[0009] In some embodiments, the stirring speed of the mixing is 500r / min~700r / min, and the mixing time is 10min~30min; The reaction temperature is 60°C~80°C, and the reaction time is 24h~48h; The drying method includes one or more of vacuum drying and freeze drying.

[0010] On the other hand, the present application also provides an agricultural preparation, which includes the above-mentioned nanozyme.

[0011] In some embodiments, the agricultural preparation includes a seed coating agent, and the nanozyme accounts for 5% to 9% of the weight of the seed coating agent; The seed coating agent comprises the following components by weight: 2 to 10 parts of pesticide; 5 to 9 parts of the above-mentioned nanozyme; Thickener 1.5~3 parts; 0.75 to 1.5 parts of bio-based polymer; 0.1~0.5 parts of organic acid; 3 to 5 parts of organic nitrogen compound; 0.5~1.5 parts of mineral materials; 0.1~0.3 parts of film-forming agent.

[0012] In some embodiments, the organic acid comprises one or more of salicylic acid, citric acid, ascorbic acid and tartaric acid.

[0013] In some embodiments, the mineral material includes one or more of attapulgite, diatomaceous earth, bentonite and sepiolite.

[0014] On the other hand, the present application also provides the use of the above agricultural formulation in improving the germination rate of seeds and the growth quality of seedlings.

[0015] In some embodiments, the seeds include one or more of soybean, wheat, corn, rapeseed, peanut, cotton, cucumber, tomato and carrot.

[0016] The raw materials for preparing the nanozyme of the present application include specific components with a specific ratio relationship, wherein the metal salt provides the active center and structural basis of the nanozyme, that is, the presence of metal ions can provide an active center, giving the nanozyme catalytic activity, similar to the catalytic effect of metal ions in the active site in natural enzymes; the surfactant controls the size and dispersibility of the nanozyme, and the acidic reagent adjusts the reaction conditions. The three cooperate with each other to provide a guarantee for the successful preparation and performance optimization of the nanozyme. Each component is coordinated and coordinated through a specific ratio relationship, and the nanomaterial modified by the surfactant has good water solubility and can be stably present in the form of a high concentration of colloid. When subsequently applied to seedling cultivation, it has the effect of promoting growth, enhancing stress resistance, increasing yield and improving quality for crops. In addition, the nanozyme itself has a bright blue color, so it can replace the highly toxic organic dyes commonly used in seed dressing agents on the market in a green way. BRIEF DESCRIPTION OF THE DRAWINGS

[0017] Figure 1 This is an optical microscope image of the agricultural preparation in Example 1;

[0018] Figure 2 It is a digital image of the agricultural preparation in Example 1;

[0019] Figure 3 This is the phenotype diagram of the agricultural preparation in Example 1;

[0020] Figure 4 The bar graph of the effect of the agricultural preparation on soybean germination rate in Example 1;

[0021] Figure 5 This is a diagram showing the effect of the agricultural preparation on soybean germination in Example 1;

[0022] Figure 6 This is a bar graph showing the effect of the agricultural preparation on soybean sprout length in Example 1;

[0023] Figure 7 This is a digital image showing that the agricultural formulation in Example 1 promotes soybean growth;

[0024] Figure 8 The effect of the agricultural preparation in Example 1 on the height of soybean plants;

[0025] Fig. 9 The effect of the agricultural preparation in Example 1 on the root length of soybean plants;

[0026] Fig.10 The effect of the agricultural preparation in Example 1 on the fresh weight of soybean plants;

[0027] Fig.11 This is a digital image of the effect of the agricultural formulation in Example 1 on the resistance of soybean plants to salt stress;

[0028] Fig.12This is a bar graph showing the effect of the agricultural formulation in Example 1 on the chlorophyll content of soybean plants after salt stress. DETAILED DESCRIPTION

[0029] For ease of understanding of the present application, the present application will be described more fully below, and preferred embodiments of the present application are provided. However, the present application can be implemented in many different forms and is not limited to the embodiments described herein. On the contrary, the purpose of providing these embodiments is to make the understanding of the disclosure of the present application more thorough and comprehensive.

[0030] The implementation of this application is described in detail below in combination with some implementation methods and examples. This example is implemented based on the technical solution of this application, and provides a detailed implementation method and specific operation process, but the protection scope of this application is not limited to the following examples.

[0031] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as those commonly understood by those skilled in the art to which this application belongs. The terms used herein in the specification of this application are only for the purpose of describing specific embodiments and are not intended to limit this application.

[0032] the term

[0033] Unless otherwise specified or incompatible herewith, the terms and phrases used herein shall have the following meanings:

[0034] In the present application, "plurality", "multiple" and the like, unless otherwise specified, refer to a number greater than or equal to 2. For example, "one or more" means one or greater than or equal to two.

[0035] In the present application, the terms "further", "particularly" and the like are used for descriptive purposes to indicate differences in content, but should not be construed as limiting the scope of protection of the present application.

[0036] In the present application, the technical features described in an open manner include closed technical solutions composed of the listed features, and also include open technical solutions containing the listed features.

[0037] In the present application, when it comes to a numerical interval (i.e., a numerical range), unless otherwise specified, the distribution of the optional numerical values ​​in the numerical interval is considered to be continuous, and includes the two numerical endpoints (i.e., the minimum value and the maximum value) of the numerical interval, and each numerical value between the two numerical endpoints. Unless otherwise specified, when the numerical interval only refers to the integers in the numerical interval, it includes the two endpoint integers of the numerical range, and each integer between the two endpoints, which is equivalent to directly listing each integer. When multiple numerical ranges are provided to describe features or characteristics, these numerical ranges can be combined. In other words, unless otherwise specified, the numerical range disclosed herein should be understood to include any and all sub-ranges included therein. The "numerical value" in the numerical interval can be any quantitative value, such as a number, a percentage, a ratio, etc. "Numerical interval" allows broadly including numerical interval types such as percentage intervals, ratio intervals, and ratio intervals.

[0038] In this application, unless otherwise specified, the temperature parameter is allowed to be either a constant temperature treatment or to vary within a certain temperature range. It should be understood that the constant temperature treatment allows the temperature to fluctuate within the accuracy range controlled by the instrument. Fluctuations within the range of ±5°C, ±4°C, ±3°C, ±2°C, and ±1°C are allowed.

[0039] In this application, when referring to the unit of a data range, if there is a unit only after the right endpoint, it means that the units of the left and right endpoints are the same. For example, 2h~5h means that the units of the left endpoint "2" and the right endpoint "5" are both h (hours).

[0040] As an environmentally friendly catalytic material, nanozymes have a size range of nanometers and have enzymatic reaction kinetics and specificity similar to natural enzymes. They are widely used in biology, medicine and other fields. In the field of agriculture, nanozymes are expected to solve the problems of pesticide damage and abiotic stress caused by traditional seed coating agents through the seed coating technology of integrated pesticide and fertilizer, and improve the effective utilization rate of pesticides and promote the healthy growth of seeds. First, nanozymes can significantly improve the nutrient absorption efficiency of plant roots. Applying nanozymes to plant roots can enhance the plant's ability to absorb nutrients in the soil, thereby increasing the growth rate and yield of plants. Secondly, nanozymes improve the seed's tolerance to pesticides by catalyzing reactive oxygen, thereby alleviating the degree of seed damage. In addition, nanozymes help improve the stress resistance of plants. Plants are easily damaged when facing environmental stresses, such as drought, extreme temperatures, salinity, heavy metals, etc. Nanozymes improve the plant's stress resistance by regulating the plant's metabolic pathways and signal transduction pathways, enabling it to better adapt to harsh environmental conditions. The potential application of nanozymes to promote crop growth and development and increase yields has brought new breakthroughs in the high-quality production of agricultural products. More importantly, nanozymes have many advantages. Their preparation process is simple, their chemical properties are stable, their catalytic reactions have good repeatability, and they have strong environmental tolerance and micro-fertilizer effects, making them particularly suitable for large-scale industrial production.

[0041] In addition, the seed coating agent uses film-forming materials such as bio-based polymers and thickeners, and self-assembles through organic acid cross-linking to form a "biofilm hydrogel" with a three-dimensional network structure. This biofilm is highly sensitive to changes in the external environment and can respond to environmental stimuli (such as pH, temperature, humidity, and salinity) through volume swelling and contraction, thereby intelligently controlling the release rate of fertilizers and improving the utilization rate of fertilizers. It also has the effect of retaining water and improving soil quality. However, the characteristics of nano seed coating agents and their preparation methods in traditional technologies are only reflected at the nanoscale. The components of the seed coating agent have problems such as poor biodegradability and the environmental pollution caused by small molecule surfactants, and its functions are relatively single.

[0042] Based on this, one embodiment of the present application provides a nanozyme, and the raw materials for preparing the nanozyme include, by weight: 1 to 6 parts of metal salt; 0.5 to 4 parts of surfactant; Acidic reagent 1 part to 1.04 parts; The above-mentioned metal salt includes one or more of iron salt, manganese salt, calcium salt, zinc salt and copper salt; the metal organic framework material of the above-mentioned nanozyme is Prussian blue.

[0043] Understandably, the research results fully confirm that as a typical metal-organic framework material, the crystal structure of Prussian blue nanoparticles provides them with a stable skeleton, enabling them to maintain stable catalytic activity under various environmental conditions. At the same time, its surface has been finely chemically modified, and can effectively bind to target molecules (such as superoxide anions, hydrogen peroxide, and hydroxyl radicals, etc.), thereby accelerating their decomposition or transformation, and has excellent antioxidant enzyme activity (mainly superoxide dismutase-like, catalase-like, and scavenging hydroxyl radicals). This property helps to improve the ability of crops to resist abiotic stress. In addition, the metal elements doped in Prussian blue derivative nanozymes have the effects of promoting growth, enhancing stress resistance, increasing yield and improving quality for crops. Among them, manganese can act as an activator of various enzymes in plants, participate in photosynthesis and respiration, regulate the ion balance in plants, and improve the adaptability of plants to environmental stresses such as drought and salinity. Zinc is an indispensable element for plant growth and development. It participates in crop photosynthesis, respiration, nitrogen metabolism, hormone synthesis and plant growth. Zinc can promote crop photosynthesis, is conducive to the accumulation of organic matter, and improves crop yield and stress resistance. Copper participates in plant photosynthesis and respiration, catalyzes redox reactions, promotes the metabolism and synthesis of carbohydrates and proteins, improves the stability of chlorophyll, and enhances the cold and drought resistance of plants. Iron is one of the indispensable trace elements in the growth and development of plants. It mainly promotes chlorophyll synthesis, participates in respiration, regulates plant growth and development, and improves stress resistance. Iron ions can catalyze the synthesis of chlorophyll, make plant leaves appear green, and participate in the transportation and release of oxygen, as well as the activity catalysis of multiple enzymes. Calcium is an essential trace element for some lower plants and higher plants. In higher plants, calcium is a metal cofactor of urease, which is essential for nitrogen metabolism. Calcium also participates in the activity regulation of other enzymes and promotes the normal growth and development of plants.

[0044] In some embodiments, the surfactant comprises tea seed cake extract.

[0045] In some embodiments, the acidic agent includes citric acid.

[0046] The raw materials for preparing the nanozyme of the present application include specific components with a specific ratio relationship, wherein the metal salt provides the active center and structural basis of the nanozyme, that is, the presence of metal ions can provide an active center, giving the nanozyme catalytic activity, similar to the catalytic effect of metal ions in the active site in natural enzymes; the surfactant controls the size and dispersibility of the nanozyme, and the acidic reagent adjusts the reaction conditions. The three cooperate with each other to provide a guarantee for the successful preparation and performance optimization of the nanozyme. Each component is coordinated and coordinated through a specific ratio relationship, and the nanomaterial modified by the surfactant has good water solubility and can be stably present in the form of a high concentration of colloid. When subsequently applied to seedling cultivation, it has the effect of promoting growth, enhancing stress resistance, increasing yield and improving quality for crops.

[0047] One embodiment of the present application provides a method for preparing a nanozyme, comprising the following steps: mixing, reacting and drying the above-mentioned raw materials for preparing the nanozyme to prepare the nanozyme.

[0048] In some embodiments, the stirring speed of the mixing is 500 r / min-700 r / min, and the mixing time is 10 min-30 min.

[0049] In some embodiments, the reaction temperature is 60° C. to 80° C., and the reaction time is 24 h to 48 h.

[0050] In some embodiments, the drying method includes one or more of vacuum drying and freeze drying.

[0051] The preparation method of the above-mentioned nanozyme of the present application has a simple process and low cost. Using the nanozyme as a seed coating material can not only reduce the risk of phytotoxicity of traditional pesticides to seeds, but also significantly improve the germination rate of seeds and the growth quality of seedlings.

[0052] One embodiment of the present application provides an agricultural preparation, which includes the above-mentioned nanozyme.

[0053] In some of the embodiments, the agricultural preparation includes a seed coating agent, and the nanozyme accounts for 5% to 9% of the weight of the seed coating agent.

[0054] In some embodiments, the seed coating agent comprises the following components by weight: 2 to 10 parts of pesticide; Nanozyme 5 to 9 parts; Thickener 1.5~3 parts; 0.75 to 1.5 parts of bio-based polymer; 0.1~0.5 parts of organic acid; 3 to 5 parts of organic nitrogen compound; 0.5~1.5 parts of mineral materials; 0.1~0.3 parts of film-forming agent.

[0055] The above-mentioned seed coating agent combines the nanozymes of the present application with pesticides, thickeners, bio-based polymers, organic acids, organic nitrogen compounds, mineral materials and film-forming agents. By controlling the specific ratios of each preparation raw material, the components work synergistically, which promotes the addition of nanozymes. It can not only achieve green and efficient disease prevention and control, but also provide nutrients for crops, promote crop growth and enhance their stress resistance, so that the seed coating agent has significant economic and ecological benefits, and has broad application prospects in the field of modern agriculture.

[0056] It should be noted that the value range of pesticide is "2 parts to 10 parts", which means the minimum and maximum values ​​of the range of 2 parts to 10 parts, and every value between the minimum and maximum values. Specific examples include but are not limited to the point values ​​in the embodiment and the following point values: 2 parts, 3 parts, 4 parts, 5 parts, 6 parts, 7 parts, 8 parts, 9 parts, 10 parts; or a range consisting of any two of these values, including, as an example, 3 parts to 10 parts.

[0057] The value range of nanozyme is "5 parts to 9 parts", that is, the minimum and maximum values ​​of the range of 5 parts to 9 parts, and every value between the minimum and maximum values. Specific examples include but are not limited to the point values ​​in the embodiment and the following point values: 5 parts, 6 parts, 7 parts, 8 parts, 9 parts; or a range composed of any two of these values, as an example, including: 6 parts to 9 parts.

[0058] The range of the thickener is "1.5 parts to 3 parts", which means the minimum and maximum values ​​of the range of 1.5 parts to 3 parts, and every value between the minimum and maximum values. Specific examples include but are not limited to the point values ​​in the embodiments and the following point values: 1.5 parts, 1.6 parts, 1.7 parts, 1.8 parts, 1.9 parts, 2.0 parts, 2.1 parts, 2.2 parts, 2.3 parts, 2.4 parts, 2.5 parts, 2.6 parts, 2.7 parts, 2.8 parts, 2.9 parts, 3.0 parts; or a range consisting of any two of these values, as an example, including: 2 parts to 3 parts.

[0059] The value range of bio-based polymer is "0.75 parts to 1.5 parts", that is, the minimum and maximum values ​​of the range of 0.75 parts to 1.5 parts, and each value between the minimum and maximum values. Specific examples include, but are not limited to, the point values ​​in the embodiments and the following point values: 0.75 parts, 0.85 parts, 0.95 parts, 1.05 parts, 1.15 parts, 1.25 parts, 1.35 parts, 1.45 parts or 1.5 parts; or a range composed of any two of these values, as an example, including: 1 part to 1.5 parts.

[0060] The value range of organic acid is "0.1 part to 0.5 part", which means the minimum and maximum values ​​of the range of 0.1 part to 0.5 part, and every value between the minimum and maximum values. Specific examples include but are not limited to the point values ​​in the embodiment and the following point values: 0.1 part, 0.2 part, 0.3 part, 0.4 part, 0.5 part; or a range consisting of any two of these values, as an example, including: 0.1 part to 0.4 part.

[0061] The value range of organic nitrogen compound is "3 parts to 5 parts", that is, the minimum and maximum values ​​of the range of 3 parts to 5 parts, and every value between the minimum and maximum values. Specific examples include, but are not limited to, the point values ​​in the embodiments and the following point values: 3.0 parts, 3.1 parts, 3.2 parts, 3.3 parts, 3.4 parts, 3.5 parts, 3.6 parts, 3.7 parts, 3.8 parts, 3.9 parts, 4.0 parts, 4.1 parts, 4.2 parts, 4.3 parts, 4.4 parts, 4.5 parts, 4.6 parts, 4.7 parts, 4.8 parts, 4.9 parts, 5.0 parts; or a range composed of any two of these values, as an example, including: 3 parts to 4.5 parts.

[0062] The range of the film-forming agent is "0.1 part to 0.3 part", which means that the minimum and maximum values ​​of the range of 0.1 part to 0.3 part, and each value between the minimum and maximum values, are taken. Specific examples include, but are not limited to, the point values ​​in the embodiment and the following point values: 0.1 part, 0.2 part, 0.3 part; or a range consisting of any two of these values, including, as an example, 0.1 part to 0.2 part.

[0063] In some embodiments, the following components are included by weight:

[0064] 5 to 10 parts of pesticide;

[0065] Nanozyme 5 to 9 parts;

[0066] 1.5 to 2 parts of thickener;

[0067] 0.75 to 1 part of bio-based polymer;

[0068] 0.1 to 0.25 parts of organic acid;

[0069] 3 to 5 parts of organic nitrogen compound;

[0070] 0.5~1 part of mineral materials;

[0071] 0.1~0.3 parts of film-forming agent.

[0072] By further controlling the components of the seed coating agent within the above-mentioned ratio range, the risk of traditional pesticide damage to seeds is lower, and the seed germination rate and seedling growth quality are better.

[0073] In some embodiments, the pesticides include one or more of fungicides and insecticides.

[0074] In some embodiments, the fungicide comprises one or more of pyraclostrobin, iprodione, boscalid and prothioconazole.

[0075] In some embodiments, the insecticide comprises one or more of cyhalothrin, emamectin benzoate, clothianidin and thiamethoxam.

[0076] In some embodiments, the thickener comprises one or more of guar gum, pectin, gelatin and starch.

[0077] In some embodiments, the bio-based polymer includes one or more of γ-polyglutamic acid, polylactic acid, polyhydroxyalkanoate and chitin.

[0078] In some embodiments, the organic acid comprises one or more of salicylic acid, citric acid, ascorbic acid and tartaric acid.

[0079] In some embodiments, the organic nitrogen compound includes one or more of urea, amino acid, polyacrylamide and formamide.

[0080] In some embodiments, the mineral material includes one or more of attapulgite, diatomaceous earth, bentonite and sepiolite.

[0081] In some embodiments, the film-forming agent includes one or more of polyvinyl alcohol, polyacrylate, chitosan, sodium carboxymethyl cellulose, silicate, natural plant oil, ethyl cellulose, sodium alginate, polyacrylic acid, starch and lignin.

[0082] One embodiment of the present application also provides a method for preparing the above-mentioned seed coating agent, comprising the following steps: mixing the components of the above-mentioned seed coating agent to obtain the seed coating agent.

[0083] One embodiment of the present application also provides a use of the above agricultural formulation in improving the germination rate of seeds and the growth quality of seedlings.

[0084] In some embodiments, the seeds include one or more of soybean, wheat, corn, rapeseed, peanut, cotton, cucumber, tomato and carrot.

[0085] The agricultural preparation of the present application has many advantages, such as improving the efficacy of medicine and fertilizer, enhancing the disease resistance and stress resistance of seeds, reducing the use of pesticides and fertilizers, improving the soil environment, improving the quality of agricultural products, and environmental protection advantages, through the efficient catalytic effect of nanozymes and the synergistic effect of medicines and fertilizers. It effectively promotes crop growth and enhances the stress resistance of crops, making the agricultural preparation have significant economic and ecological benefits, and has broad application prospects in the field of modern agriculture.

[0086] In order to make the purpose, technical solutions and advantages of the present application more concise and clear, the present application is described with the following specific embodiments, but the present application is by no means limited to these embodiments. The embodiments described below are only preferred embodiments of the present application and can be used to describe the present application, and cannot be understood as limiting the scope of the present application. It should be pointed out that any modifications, equivalent substitutions and improvements made within the spirit and principles of the present application should be included in the protection scope of the present application.

[0087] In order to better illustrate the present application, the present application is further described below in conjunction with the examples. The experimental methods used in the following examples are conventional methods unless otherwise specified; the reagents, materials, etc. used in the following examples are all commercially available unless otherwise specified.

[0088] Example 1

[0089] (1) Preparation of Prussian blue derivative nanozymes: Nanozymes are composed of metal salts, surfactants and acidic reagents. The specific method is as follows: 1 part of surfactant (tea seed cake extract), 1 part of acidic reagent (citric acid), 2 parts of iron salt (potassium ferrocyanide), and 0.9 parts of manganese salt, zinc salt, copper salt, and calcium salt (specifically manganese chloride, zinc nitrate, copper chloride, and calcium chloride) are added to 50 mL of deionized water containing 0.02 parts of acidic reagent, stirred at 600 r / min for 10 minutes at room temperature, placed in a vacuum drying oven at 80°C for 20 hours, washed the reaction product with deionized water, then added anhydrous ethanol and centrifuged, repeated three times, and freeze-dried to obtain nanozyme powder. Please refer to Table 1 for details.

[0090] (2) Preparation of seed coating agent: According to the weight proportion, 10 parts of fludioxonil and 0.1 parts of salicylic acid are mixed, and the mixture is ground to a particle size of 80 nm to 100 nm, and 3 parts of urea and 0.5 parts of attapulgite are added and mixed evenly. Finally, 1.5 parts of guar gum, 0.75 parts of γ-polyglutamic acid, 5 parts of nanozymes, and 0.3 parts of a film-forming agent (starch) are added and stirred evenly to obtain a seed coating agent.

[0091] (3) Preparation of biofilm-based seed coating: Soybean seeds are immersed in the above-mentioned seed coating agent for 1 minute, taken out and dried to obtain seeds with a coating.

[0092] Embodiment 2~6

[0093] The preparation methods of the seed coating agents in Examples 2 to 6 are basically the same as those in Example 1, except that the components of the seed coating agents are different, as shown in Tables 1 and 2 for details.

[0094] The other steps and conditions are the same as those in Example 1.

[0095] Example 7

[0096] The preparation methods of the seed coating agent in Example 7 and Example 1 are basically the same, except that the ratio of the raw materials for preparing the nanozyme is different, specifically the ratio of the metal salt is different. In Example 7, 1 part of iron salt (specifically potassium ferrocyanide), and 0.1 parts of manganese salt, zinc salt, copper salt, and calcium salt (specifically manganese chloride, zinc nitrate, copper chloride, and calcium chloride) are used, see Table 1 to Table 2 for details.

[0097] The other steps and conditions are the same as those in Example 1.

[0098] Example 8

[0099] The preparation methods of the seed coating agent in Example 8 and Example 1 are basically the same, except that the ratio of the raw materials for preparing the nanozyme is different, specifically the ratio of the metal salt is different. In Example 8, 2 parts of iron salt (specifically potassium ferrocyanide), and 0.3 parts of manganese salt, zinc salt, copper salt, and calcium salt (specifically manganese chloride, zinc nitrate, copper chloride, and calcium chloride) are used, see Table 1 to Table 2 for details.

[0100] The other steps and conditions are the same as those in Example 1.

[0101] Comparative Example 1

[0102] The preparation method of the seed coating agent in Comparative Example 1 is basically the same as that in Example 1, except that the ratio of the raw materials for preparing the nanozyme is different, specifically the ratio of the acidic reagent is different, which is 0.005 parts of citric acid in Comparative Example 1, see Tables 1 and 2 for details.

[0103] The other steps and conditions are the same as those in Example 1.

[0104] Comparative Example 2

[0105] The preparation methods of the seed coating agents in Comparative Example 2 and Example 1 are basically the same, except that the components of the seed coating agents are different and nanozymes are not introduced, see Table 2 for details.

[0106] The other steps and conditions are the same as those in Example 1.

[0107] Comparative Example 3

[0108] The preparation methods of the seed coating agents in Comparative Example 3 and Example 1 are basically the same, except that the components of the seed coating agents are different, specifically the content of organic acid (salicylic acid). Comparative Example 3 contains 0.6 parts of salicylic acid, see Table 1 to Table 2 for details.

[0109] The other steps and conditions are the same as those in Example 1.

[0110] Table 1

[0111]

[0112] Table 2

[0113]

[0114] test:

[0115] 1. Film-forming time detection of different formulas

[0116] (1) Experimental methods

[0117] When a good suspended seed coating agent is used to coat seeds, the liquid can quickly solidify into a film on the surface of the seeds to form a protective film to defend against pests and diseases. Film-forming property is an important test indicator in the application process of suspended seed coating agents. Test method: Weigh 50g of seeds (accurate to 1g) in a culture dish, use a syringe to draw 1g of the sample and inject it into the culture dish, cover it and turn it over for 5 minutes, open the lid, spread the coated seeds flat to form a film, use a glass rod to gently move the seeds, and observe the surface of the seeds. If the seed coating agent on the surface of all seeds solidifies into a film, the film-forming property is qualified. Record the film-forming time of each formula separately.

[0118] (2) Data Analysis

[0119] The test results are shown in Table 3. It can be clearly seen that the formulation of Example 1 has a fast film-forming property.

[0120] Table 3

[0121]

[0122] 2. Coating uniformity test of different formulations

[0123] (1) Experimental methods

[0124] Randomly select 100 coated seeds with qualified film-forming properties from the coated seeds made with different formulas, place them in 25 stoppered centrifuge tubes (4 seeds in each centrifuge tube), accurately add 2.0~5.0mL ethanol solution (to ensure the absorbance is within the linear range) with a pipette, cover and soak for 1h, shake and extract for 15min, and let stand or centrifuge to obtain a clear solution. Using ethanol solution as a reference, measure its absorbance A at the maximum absorption wavelength of the extract. Arrange the 25 measured absorbance data from small to large, and calculate the average absorbance value A. Coating uniformity X(%)=n / 25 100%, where n is the number of centrifuge tubes with absorbance A in the range of 0.7A~1.3A; 25 is the total number of centrifuge tubes.

[0125] (2) Data Analysis

[0126] The test results are shown in Table 4. It can be clearly seen that the coating effect of the formulation in Example 1 is the most uniform.

[0127] Table 4

[0128]

[0129] 3. Shedding rate test of different formulas

[0130] (1) Experimental methods

[0131] Weigh 10g (accurate to 0.02g) of two portions of coated seeds to be tested for film-forming properties and place them in conical flasks. Accurately add 100mL of ethanol to one portion, stopper it and place it in an ultrasonic cleaner for 10 minutes to fully dissolve the seed coating agent on the surface of the seeds. Take it out and let it stand for 10 minutes. Take 5mL of the solution and place it in a 50mL volumetric flask, dilute it to the scale with ethanol, shake it well, and obtain solution A. Place the other portion of coated seeds in an oscillator, oscillate for 10 minutes, then carefully place the seeds in another conical flask and treat them according to the method of solution A to obtain solution B. Using ethanol as a reference, measure its absorbance at the maximum absorption wavelength of the solution. The shedding rate after coating X(%)=(A 0 m 1 -A 1 m 0 ) / A 0 m 1 100%, where m 0 is the mass of the coated seeds weighed to prepare solution A, g; m 1 A is the mass of the coated seeds weighed to prepare solution B, g; 0 is the absorbance of solution A; A 1 is the absorbance A of solution B.

[0132] (2) Data Analysis

[0133] The test results are shown in Table 5, and it can be clearly seen that the formulation of Example 1 has excellent anti-shedding performance.

[0134] Table 5

[0135]

[0136] 4. Detection of pesticide damage to soybeans by different formulations

[0137] (1) Experimental methods

[0138] The coated seeds of the embodiment and the comparative example were placed in a 9 cm culture dish, 10 seeds were placed in each dish, and 3 replicates were set for each treatment group. During the culture, the humidity environment was maintained at 60%, and the germination rate was counted after three days. The uncoated seeds were used as blank controls.

[0139] (2) Data Analysis

[0140] The test results are shown in Table 6, which clearly shows that the formulation in Example 1 did not cause phytotoxicity to soybeans.

[0141] Table 6

[0142]

[0143] 5. Test on the effect of different formulas on soybean root rot prevention and control

[0144] (1) Experimental methods

[0145] Prepare the soil with Pythium aphanidermatum and put it in pots, then coat the seeds (such as Figure 3 ) were sown into flower pots, 10 seeds were sown in each pot, and 4 pots were set for each treatment group. When the seeds germinated and grew to the 3-leaf stage, the disease rate and disease level were investigated in the pots, and the control effect was calculated. Among them, the uncoated seeds were blank controls.

[0146] (2) Data Analysis

[0147] The test results are shown in Table 7, which clearly shows that the formula of Example 1 has an excellent effect on preventing and controlling soybean root rot.

[0148] Table 7

[0149]

[0150] 6. Effect of Seed Coating Agent on Soybean Germination Rate and Bud Length in Example 1

[0151] (1) Experimental methods

[0152] The coated seeds prepared in Example 1 were placed on filter paper, 25 seeds in each group, for a total of 3 groups, with uncoated seeds as blank controls. The humidity was maintained at 60%-90%, and the germination rate and bud length were counted after 3 days.

[0153] (2) Data Analysis

[0154] The test results are as follows Figure 4 and Figure 5 As shown in the figure, it can be clearly seen that seed coating has no adverse effect on soybean germination, and the germination rate is not much different, but the effect of promoting seed bud growth is obvious (such as Figure 6 ).

[0155] 7. Effect of Seed Coating Agent on Soybean Growth in Example 1

[0156] (1) Experimental methods

[0157] The coated seeds were sown in the soil, and the control group was uncoated seeds. The potted plants were placed in a light incubator for cultivation. After one week, the root length, plant height and fresh weight of each plant were observed and recorded.

[0158] (2) Data Analysis

[0159] The test results are as follows Figure 7 As shown, it can be seen that the seed coating agent of Example 1 has a promoting effect on soybean growth, which is specifically manifested in the promotion of root length, plant height and fresh weight (such as Figure 8 , Fig. 9 , Fig.10 ).

[0160] 8. Example 1: Effect of Seed Coating Agent on Plant Salt Stress Resistance

[0161] (1) Experimental methods

[0162] The coated seeds were sown in the soil, and the control group was uncoated seeds. The potted plants were placed in a light incubator and treated with 0.1% NaCl solution to simulate salt stress. After one week, the growth of each plant was observed and recorded, and the chlorophyll content of its leaves was measured.

[0163] (2) Data Analysis

[0164] The test results are as follows Fig.11 As shown, it can be seen that the seed coating agent of Example 1 has a promoting effect on the salt stress resistance of soybeans, and the chlorophyll content is also higher than that of uncoated seeds (such as Fig.12 ).

[0165] From the above Tables 3 to 7 and Figure 1 to Figure 12 The results show that, Figure 1This is an optical microscope image of the agricultural preparation in Example 1; Figure 2 It is a digital image of the agricultural preparation in Example 1; Figure 3 This is the phenotype diagram of the agricultural preparation in Example 1; Figure 4 The bar graph of the effect of the agricultural preparation on soybean germination rate in Example 1; Figure 5 This is a diagram showing the effect of the agricultural preparation on soybean germination in Example 1; Figure 6 This is a bar graph showing the effect of the agricultural preparation on soybean sprout length in Example 1; Figure 7 This is a digital image showing that the agricultural formulation in Example 1 promotes soybean growth; Figure 8 The effect of the agricultural preparation in Example 1 on the height of soybean plants; Fig. 9 The effect of the agricultural preparation in Example 1 on the root length of soybean plants; Fig.10 The effect of the agricultural preparation in Example 1 on the fresh weight of soybean plants; Fig.11 This is a digital image of the effect of the agricultural formulation in Example 1 on the resistance of soybean plants to salt stress; Fig.12It is a bar graph of the chlorophyll content of soybean plants after salt stress by agricultural preparations in Example 1. The above agricultural preparations have the following effects: 1. Solve the problem of pesticide damage: Nanozymes, as an active oxygen catalytic material, have the advantages of high efficiency, low toxicity and repeatable catalysis, and can effectively degrade the free radicals produced by pesticide stress and reduce their toxic effects on biological systems. In the field of agriculture, the application of nanozymes can not only reduce the risk of pesticide damage to seeds by traditional pesticides, but also significantly improve the germination rate of seeds and the growth quality of seedlings. This technology provides a safer and more environmentally friendly solution for agricultural production, which helps to improve crop yield and quality. 2. Promote crop growth: Through the application of nanotechnology, drug and fertilizer ingredients can be more effectively attached to the surface of seeds, forming a lasting protective layer, continuously inhibiting the invasion and spread of pests and diseases, and providing necessary nutrient support at the same time. Nanozymes, with their small size and large specific surface area, can contact the surface of seeds more efficiently, significantly improving the utilization rate of drug and fertilizer effects. In addition, the in situ catalytic oxygen production characteristics of nanozymes provide sufficient oxygen for rhizosphere microorganisms, further promoting the healthy growth of crops. This innovative technology brings more efficient and environmentally friendly solutions to agricultural production and increases crop yields. 3. Enhance seed disease resistance: Nanozyme coatings achieve effective protection of plants by directly destroying pathogens or enhancing the plant's own immune system (such as increasing the activity of antioxidant enzymes in crops). This technology can significantly improve the disease resistance of seeds and reduce the risk of crops being attacked by pests and diseases, thereby increasing crop yield and quality. Nanozyme coatings provide an efficient and sustainable solution for plant disease prevention and control, with broad application prospects. 4. Improve stress resistance: The pesticide components in seed coatings can effectively prevent and control pests and diseases. At the same time, the introduction of nanozymes enables them to exert multiple enzyme activities, efficiently catalyze active oxygen, and further improve the stress resistance of crops, whether in response to biological or abiotic stress. The unique structure of nanozymes can also enhance the adaptability of crops to adverse environments and provide crops with a more comprehensive protection mechanism. This technology not only improves the stress resistance of crops, but also reduces the threat of pests and diseases, realizes the synergistic effect of pesticides and fertilizers, and optimizes agricultural production results. 5. Reduce the amount of pesticides used: Nanozymes, due to their high specific surface area and porosity, help to achieve the sustained release effect of active ingredients in pesticides, thereby significantly improving the utilization rate of pesticides. At the same time, nanozymes can improve the dispersibility and stability of pesticides, enhance their solubility and mobility in the soil, and promote the effective use of pesticides by plants. Through this mechanism, not only the amount of pesticides used is reduced, but also the risk of environmental pollution is reduced, achieving a more environmentally friendly and efficient agricultural production goal. 6. Improve the yield and quality of agricultural products: The agricultural preparations of the embodiments of the present application significantly improve the yield and quality of crops by promoting crop growth and enhancing stress resistance. In addition, since the amount of pesticides used is effectively reduced, the safety of agricultural products is improved, thereby enhancing their market competitiveness.This innovative technology not only optimizes agricultural production efficiency, but also provides strong support for sustainable agricultural development. 7. Environmental protection and cost advantages: Nanoenzyme coating technology has environmental advantages, which can reduce the loss and volatilization of pesticides and reduce pollution to the environment. At the same time, this technology can also improve the environmental behavior and biosafety of pesticides and enhance the safety of chemical control to the ecological environment. The raw materials of the seed coating are widely available, the preparation process is simple, and it is easy to mass-produce.

[0166] In summary, agricultural preparations have many advantages, such as improving the efficacy of medicine and fertilizer, enhancing the disease resistance and stress resistance of seeds, reducing the use of pesticides and fertilizers, improving the soil environment, improving the quality of agricultural products, and environmental protection advantages, through the efficient catalytic effect of nanozymes and the synergistic effect of pesticides and fertilizers. It effectively promotes crop growth and improves the stress resistance of crops, making the agricultural preparations have significant economic and ecological benefits, and has broad application prospects in the field of modern agriculture.

[0167] The technical features of the above-described embodiments may be arbitrarily combined. To make the description concise, not all possible combinations of the technical features in the above-described embodiments are described. However, as long as there is no contradiction in the combination of these technical features, they should be considered to be within the scope of this specification.

[0168] The above-described embodiments only express several implementation methods of the present application, and the descriptions thereof are relatively specific and detailed, but they cannot be construed as limiting the scope of the patent application. It should be pointed out that, for a person of ordinary skill in the art, several variations and improvements can be made without departing from the concept of the present application, and these all belong to the protection scope of the present application. Therefore, the protection scope of the patent application shall be subject to the attached claims.

Claims

1. A nanozyme, characterized in that: The raw materials for preparing the nanozyme include, by weight: 1 to 6 parts of metal salt; 0.5 to 4 parts of surfactant; 1 part to 1.04 parts of acid reagent; The metal salt includes one or more of iron salt, manganese salt, calcium salt, zinc salt and copper salt; the metal organic framework material of the nanozyme is Prussian blue.

2. A nanozyme according to claim 1, characterized in that: The surfactant includes tea seed cake extract; The acidic agent includes citric acid.

3. A method for preparing a nanozyme, characterized in that: The steps include: The raw materials for preparing the nanozyme as described in any one of claims 1 to 2 are mixed, reacted and dried to prepare the nanozyme.

4. A method for preparing the nanozyme according to claim 3, characterized in that: The mixing speed is 500r / min~700r / min, and the mixing time is 10min~30min; The reaction temperature is 60°C~80°C, and the reaction time is 24h~48h; The drying method includes one or more of vacuum drying and freeze drying.

5. An agricultural preparation, characterized in that: The agricultural preparation comprises the nanozyme according to any one of claims 1 to 2.

6. An agricultural formulation according to claim 5, characterized in that The agricultural preparation includes a seed coating agent, and the nanozyme accounts for 5% to 9% based on the weight of the seed coating agent; The agricultural preparation comprises the following components by weight: 2 to 10 parts of pesticide; Nanozyme 5 to 9 parts; Thickener 1.5~3 parts; 0.75 to 1.5 parts of bio-based polymer; 0.1~0.5 parts of organic acid; 3 to 5 parts of organic nitrogen compound; 0.5~1.5 parts of mineral materials; 0.1~0.3 parts of film-forming agent.

7. An agricultural formulation according to claim 6, characterized in that The organic acid includes one or more of salicylic acid, citric acid, ascorbic acid and tartaric acid.

8. An agricultural formulation according to claim 6, characterized in that The mineral material includes one or more of attapulgite, diatomaceous earth, bentonite and sepiolite.

9. Use of the agricultural preparation according to any one of claims 6 to 8 for improving the germination rate of seeds and the growth quality of seedlings.

10. A use according to claim 9, characterized in that The seeds include one or more of soybean, wheat, corn, rapeseed, peanut, cotton, cucumber, tomato and carrot.

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