Bioactive formulation of levorotatory amino acids fortified with copper and potassium for the prevention and control of frost damage in agricultural products

Abstract

This invention relates to the formulation and synthesis of a levorotatory amino acid biocomplex enriched with copper and potassium ions, which has been developed with the aim of increasing plant resistance and effectively preventing freezing in agricultural crops. The presented composition significantly increases the plant's ability to withstand critical temperatures by utilizing synergistic biological mechanisms, including inhibiting the formation and growth of ice crystals, increasing the osmotic pressure of the cell sap, stabilizing membranes and proteins, and reducing the density of ice-forming bacteria. This biocomplex is produced by the controlled reaction of 19 left-handed amino acids with soluble copper and potassium salts in an aqueous medium under precise temperature and time conditions. The final product is a stable compound, completely water-soluble and highly bioavailable. The final formulation contains approximately 17–18% left-handed amino acids, 4–5% copper ions and 22–23% potassium ions. This compound is capable of being produced on an industrial scale and is used as a specialized input in gardens, agricultural fields, greenhouses and hydroponic systems. It is applied as a foliar spray at a concentration of 6 parts per thousand and increases plant metabolism, stimulates enzyme activity, protects sensitive tissues from cold, and significantly reduces damage caused by frostbite.

Description

Description of the invention Title of the invention Bioactive formulation of levorotatory amino acids fortified with copper and potassium for the prevention and control of frost damage in agricultural crops. Technical background of the relevant invention This invention relates to the field of agriculture, plant nutrition and the production of growth-promoting biocompounds, and specifically to the formulation and synthesis of levorotatory amino acid biocomplexes enriched with copper and potassium micronutrients. This compound is designed to increase plant resistance and prevent and control climatic stresses caused by frost, as well as reduce frost damage in horticultural, field, greenhouse and hydroponic crops. Technical problem and stating the objectives of the invention Cold stress is one of the most important abiotic stresses in agriculture, causing significant damage to horticultural, field, and greenhouse crops by causing cellular damage, metabolic disorders, and yield reduction. Existing strategies for preventing and controlling cold stress often have limited effectiveness and are not capable enough to inhibit ice-forming bacteria, stabilize cell structure, and maintain plant physiological activities at critical temperatures. This invention has been developed with the aim of providing a copper and potassium enriched levorotatory amino acid biocomplex formulation. This compound is synthesized by the reaction of levorotatory amino acids with soluble copper and potassium salts in an aqueous environment under controlled temperature and time conditions, and the final product is a stable, water-soluble, and highly absorbable biocomplex. The purpose of this invention is to increase plant resistance to frost, reduce the formation and growth of ice crystals, improve cellular osmotic pressure, strengthen the antioxidant system, and reduce the density of ice-forming bacteria. The simultaneous presence of bioactive amino acids and copper-amino acid chelates improves metabolism, increases the activity of defense enzymes, and enhances plant tolerance in the period before and during cold. A description of the state of the prior art and the history of developments related to the claimed invention. Cold stress is one of the most important abiotic stresses in agriculture, usually occurring at temperatures of 0 to 10 degrees Celsius, depending on the plant species and cultivar, and causing physiological and morphological damage, including growth retardation, chlorosis, leaf scorch, cell tissue destruction, impaired germination, vascular occlusion, and reduced photosynthetic material transport. After the occurrence of cold stress, cell and vascular destruction disrupts normal growth, reduces nutrient absorption, and increases plant susceptibility to pathogens. Despite the importance of this phenomenon, many existing methods do not have sufficient ability to prevent ice crystal formation, inhibit ice-forming bacteria, and maintain plant metabolic activities at critical temperatures. In recent years, the use of amino acids, osmolytes, and micronutrients has been considered as adjuvant strategies to increase plant resistance to cold; however, most of these compounds have lacked stability, adequate absorption, or the ability to simultaneously concentrate cell sap, stabilize membrane structure, and control ice-forming bacteria. Common sources of copper and potassium also do not show optimal efficiency under cold conditions due to limited solubility or low bioavailability. Given this scientific and technical gap, there is a need to develop a compound that can synergistically provide plant nutrition, increase osmotic pressure, inhibit ice crystals, and control ice-forming bacteria. The present invention aims to address this need by introducing a biocomplex of left-handed amino acids enriched with copper and potassium. This compound is designed to: - Deliver copper to the sensitive tissues of the plant in a complexed and absorbable form, -Reduce the density of ice-nucleating bacteria, -Increase the osmotic pressure of cell sap and interstitial water by introducing amino acid and potassium osmolytes, And it provides the necessary conditions to increase the plant's resistance to frost. The synthesis process of this biocomplex involves dissolving soluble copper and potassium compounds in water and adding a mixture of 19 left-handed amino acids. The reaction is carried out at a temperature of 50 to 80 degrees Celsius for 1 to 5 hours to form a stable, bioactive, and completely water-soluble complex. The final product is sprayed on plants at a specific concentration before frost occurs. Based on prior knowledge, known mechanisms for increasing plant resistance to cold include inhibiting the growth and recrystallization of ice crystals, increasing osmotic pressure, stabilizing proteins and membranes, and enhancing the antioxidant system. The biocomplex presented in this invention, by utilizing these same principles and by simultaneously combining bioactive amino acids, short-chain peptides, and copper-amino acid chelates, provides a more comprehensive and effective function than existing compounds. The presence of copper in the complexed form, in addition to its nutritional effects, reduces the density of ice-forming bacteria and improves the activity of defense enzymes such as Cu / Zn-SOD, and plays an important role in reducing oxidative damage after cold. This set of advances demonstrates that the technology presented in this invention provides an innovative, sustainable, and efficient solution for preventing and controlling frost damage in agricultural crops, compared to previous methods and compositions. Providing a solution to an existing technical problem along with an accurate, sufficient, and integrated description of the invention Given the limitations of existing methods for preventing frost damage and the ineffectiveness of common compounds in inhibiting ice crystal formation, increasing osmotic pressure, stabilizing cell structure, and controlling ice-forming bacteria, the present invention offers a comprehensive, sustainable, and innovative solution. This solution is based on the design and synthesis of a biocomplex of levorotatory amino acids enriched with copper and potassium ions, which synergistically activates several plant defense mechanisms and significantly increases its resistance to frost damage. The proposed solution and its scientific logic is that this biocomplex, by simultaneously combining three key components including left-handed amino acids, micronutrients copper and potassium, creates a bioactive structure that is capable of: -Transfer copper in a complexed form with high bioavailability to germ cells and sensitive tissues; -Reduce the concentration of ice-forming bacteria such as Pseudomonas syringae and prevent ice nucleation; -Increase the osmotic pressure of cell sap and interstitial water through the entry of amino acid and potassium osmolytes; - Stabilize membrane structure, proteins, and vital enzymes against cold stress; - Ultimately inhibit the formation, growth, and recrystallization of ice crystals. This set of functions allows the presented combination to act as an integrated and effective agent in preventing frostbite, a capability that did not exist in previous technologies. In the product of this invention, about 17-18% by weight of the mixture of left-handed amino acids, 4-5% by weight of copper ions, and 22-23% of potassium ions are provided in the final product, respectively. The biocomplex synthesis method and production process includes the following steps: First, copper compounds are added to the mixture of left-handed amino acids. A mixture of 19 left-handed amino acids including alanine, arginine, aspartic acid, cysteine, glutamic acid, valine, glycine, histidine, isoleucine, leucine, lysine, methionine, phenylalanine, proline, serine, threonine, tryptophan, tyrosine, and glutamine is added as bioactive ligands to initiate the complexation reaction. After the copper-amino acid complexation is complete, potassium-containing compounds are added to the reaction mixture. The resulting mixture is stirred continuously for about 3 hours at 20 to 40°C to form a stable, uniform, and fully water-soluble complex. The final product is a stable, homogeneous, fully water-soluble, and rapidly absorbable liquid. Implementation example: In a working example, first 1000 grams of copper nitrate trihydrate was powdered and slowly added to 2000 ml of a solution of left-handed amino acids (including the 19 amino acids mentioned) and stirring was continued until the solution became uniform. Then 3000 grams of potassium sulfate was added to the mixture. The final mixture was stirred for 3 hours at a temperature of 20 to 40 ° C to complete the complexation reaction and obtain a stable and completely water-soluble mixture. Application and function: This biocomplex is used as a foliar spray on the leaves, stems, and trunks of plants 1 to 2 days before frost occurs at a concentration of 6 parts per thousand. Its functions include: -The presence of left-handed amino acids and potassium as fully nutritional osmolytes increases the osmotic pressure of cell sap and interstitial water. -Delaying the formation of ice crystals and changing the shape of the crystals in a way that prevents their growth; -Reducing the density of ice-forming bacteria by delivering complexed copper to sensitive tissues; -Increase cellular metabolism and stimulate enzymatic activity; -Enhance plant resistance to frost through a set of nutritional and protective mechanisms. The sum of these compounds acts as nutritional elements and greatly increases the plant's resistance to frost by increasing cellular metabolism and stimulating enzyme activity. Explanation of shapes, maps and diagrams The process block diagram is provided in the attached file. A clear and precise statement of the advantages of the claimed invention over prior inventions. Until now, no comprehensive, effective and integrated method has been presented for the prevention and control of frost damage in plants. The present invention has a set of technical, functional and structural advantages over existing compounds, methods and formulations in the field of frost prevention, which makes it an innovative and superior technology. The most important advantages of this invention are: 1. Integrated combination of three key factors in a single formulation In previous technologies, amino acids, copper compounds, and potassium sources were typically used separately or in non-synergistic formulations. The present invention combines, for the first time, levorotatory amino acids, complexed copper ions, and highly bioavailable potassium ions in a stable, homogeneous biocomplex. This integration increases efficiency and reduces the need for multiple products. 2. Increase copper bioavailability and prevent toxicity and precipitation In previous methods, copper was usually used as mineral salts which: had low bioavailability, precipitated at different pHs, and increased the possibility of toxicity or leaf burn. In this invention, copper is provided in a complexed form with left-handed amino acids which: increases its absorption several-fold, reduces precipitation and toxicity, and effectively transports it to sensitive plant tissues. 3. Simultaneous control of the three main pathways of frostbite injury Previous technologies typically targeted only one of the damage pathways (e.g., just nutrition, or just anti-icing). The present invention simultaneously inhibits three major pathways of frost damage: Inhibiting the formation and growth of ice crystals Increasing osmotic pressure and preventing intracellular freezing Reducing the density of ice-forming bacteria This synergy significantly increases the plant's resistance to critical temperatures. 4. Use of left-handed amino acids with high biological activity In many existing products, amino acids are used in the form of D / L mixtures or low-quality hydrolysates. In this invention: only left-handed amino acids (L-form, with high biological activity and effective complexing ability) are used. This leads to: increased absorption, increased osmotic pressure, and improved cold protection. 5. High stability and complete solubility in water Many existing compounds: precipitate, do not dissolve completely in water, or become unstable over time. The biocomplex of this invention: is completely stable, completely water-soluble, and has high molecular uniformity. This property is crucial for foliar application and rapid absorption. 6. Effective control of ice-forming bacteria In previous technologies, the control of ice-forming bacteria was usually achieved with: non-complexed copper compounds, or chemicals with limited effect. In this invention, the copper-amino acid chelate: has higher permeability, provides stronger antibacterial effect, and effectively reduces the possibility of ice nucleation. 7. High compatibility with plants and reduced risk of burns Due to the complexation of ions with left-handed amino acids: the risk of leaf burn is reduced, copper toxicity is controlled, and the product can be used for a wide range of garden, field, and greenhouse plants. 8. Increase metabolism and stimulate enzyme activity In addition to its protective role, this biocomplex is also a complete biostimulant and causes: increased activity of antioxidant enzymes, increased ATP production, improved photosynthesis, and accelerated regeneration of damaged tissues. This benefit is less common in previous products. 9. Fast performance and high effectiveness at critical temperatures Due to its rapid absorption and high bioavailability, this compound: becomes active shortly after foliar application, and can provide a protective effect 1 to 2 days before frost occurs. This feature is not seen in many existing products. Description of at least one implementation method for implementing the invention For the practical use of the biocomplex of left-handed amino acids enriched with copper and potassium ions, the following method can be used as a standard operating procedure. This method is designed to achieve the highest efficiency of the product in preventing frost damage and increasing plant resistance. 1. Consumption schedule Foliar spraying should be done 1 to 2 days before frost occurs. If the cold is expected to continue, repeated application is possible after 3 days. The best time for foliar spraying is early morning or evening so that the leaves can absorb it completely. For orchards, foliar spraying should be done when the buds begin to swell and continue until the flowers open. Avoid spraying on newly opened flowers. After the initial fruit formation, foliar spraying can be repeated. For crops and greenhouses, repeat every three days until the cold wave passes. 2. For gardens, 6 liters of the biocomplex product is dissolved in 1000 liters of water and sprayed (concentration 6 in 1000). For farms and greenhouses, 5 liters of the product is dissolved in 1000 liters of water and sprayed. The water used must be free of high hardness and free chlorine to maintain the stability of the complex. The spray tank must be clean and free of fertilizer or other pesticide residues. 3. Foliar spraying method The prepared solution is sprayed evenly on buds, leaves, fruit, stems, trunks, and all branches. Avoid spraying this mixture on flowers. Complete coverage of the plant surface is essential, but the solution should be prevented from running onto the leaves. In orchards, the volume of foliar spray is adjusted based on the age of the tree and the density of the shoots. 4. Mechanism of action during application After spraying, the left-handed amino acids and potassium enter the tissues as osmolytes and increase the osmotic pressure of the cell sap, to prevent intracellular freezing. And effectively delay the formation of ice crystals. The complexed copper is also rapidly absorbed and reduces the density of ice-forming bacteria, and also increases the activity of defense enzymes such as Cu / Zn-SOD, and prevents ice nucleation. In addition, the structure of the membrane and sensitive proteins are stabilized, and the resistance of tissues to cold shock is increased, and cellular metabolism is maintained at low temperatures. 5. Suitable environmental conditions for the operation: The ambient temperature during foliar spraying should be between 2 and 20 degrees Celsius. Spraying should be avoided during rain or strong winds. Rain should not occur for at least 4 to 6 hours after spraying. 6. Compatibility and Mixability Mixing with fruit set ingredients should be avoided. Avoid spraying on open flowers. By properly implementing this method, the formation of ice crystals in tissues is significantly reduced, sensitive plant tissues become resistant to cold shock, frost damage to buds, flowers, and leaves is reduced, and normal plant growth is maintained after the cold period. Practical example (for one hectare of garden, farm or greenhouse) Application method in the garden, field or greenhouse before frost occurs Before frost occurs, the biocomplex of left-handed amino acid-copper-potassium should be diluted in water and applied as a direct foliar spray on the leaves, stems, and fruits of affected plants. For one hectare of cultivated area, 5-6 liters of the biocomplex is dissolved in 1000 liters of water and the resulting solution is sprayed on all plant surfaces using a manual or motorized sprayer. This foliar spray: Rapidly delivers nutrients to plant cells, including swelling buds, newly formed fruits, plant leaves, and other sensitive plant organs, and protects the plant, buds, fruits, young branches, and leaves from damage caused by cold stress, and enables normal plant growth. For areas less than or greater than one hectare, the amount of biocomplex and water volume are adjusted accordingly. Explicit mention of the industrial application of the invention This invention has the ability to be produced on an industrial scale and can be implemented in biofertilizer production units, plant growth stimulants and specialized agricultural input production industries. The final product of this invention is a levorotatory amino acid biocomplex enriched with copper and potassium ions, which is used as an efficient and specialized compound in managing climatic stresses, especially preventing frostbite and reducing its damages. This product has the ability to be widely used in gardens, agricultural fields, greenhouses and hydroponic systems and can be used to: increase plant resistance to frostbite, reduce the formation and growth of ice crystals in plant tissues, accelerate the repair and regeneration of damaged tissues, improve the nutritional and metabolic status of the plant, and improve yield and quality of the product as a foliar spray or used in plant nutritional programs. The stable nature, complete solubility in water, and high bioavailability of the active components of this biocomplex make it perfectly suitable for mass production and use in professional and industrial agricultural operations. Industrial production method (executive example for a 6000 kg batch) For industrial production of this compound on a scale of 6000 kg per batch, the following steps are performed: Adding copper compounds: 1,000 kilograms of trihydrate copper nitrate powder is poured into an industrial mixer with a capacity of about 10,000 liters. Adding the left-handed amino acid solution: Then, 2000 liters of left-handed amino acid solution are gradually added to the tank and stirring is continued until the solution becomes uniform. Adding potassium compounds: 3000 kilograms of potassium sulfate is added to the mixture gradually and with stirring until it becomes completely homogeneous. Completion of the complexation reaction: The final mixture is stirred continuously for 3 hours at a temperature of 20 to 40°C until the complexation reaction is complete and a stable, uniform, and fully water-soluble biocomplex is obtained. This process is repeatable, scalable, and compatible with standard liquid fertilizer production equipment, enabling continuous production in industrial lines. Practical application of the product in the agricultural industry Before frost occurs, the product is used as a nutritional-preventive solution and a quick cleanser of ice-forming bacteria. The method of use in gardens, fields or greenhouses is as follows: 5 to 6 liters of biocomplex is dissolved in 1000 liters of water. The prepared solution is sprayed evenly on the leaves, stems, branches and fruits of plants. This amount is suitable for one hectare of cultivated area and for smaller or larger areas, the amount of product and water is adjusted accordingly. Effects after use: Rapid delivery of nutrients and active elements to plant cells, maintaining metabolic activity at low temperatures, reducing the formation of ice crystals and preventing their growth and recrystallization, reducing the density of ice-forming bacteria and preventing ice nucleation, and ultimately significantly reducing damage caused by frostbite.

Claims

Claim What is claimed: Claim 1) The claimed product is a bioactive formulation of left-handed amino acids enriched with copper ions and potassium ions, which is used to prevent and control frost damage in agricultural crops. Claim 2) According to claim 1, the bioactive formulation of a biocomplex of left-handed amino acids enriched with copper and potassium contains about 18-17% by weight of a mixture of 19 left-handed amino acids, 4-5% by weight of copper ions, and 23-22% by weight of potassium ions, respectively. Claim 3) The method for synthesizing a levorotatory amino acid-copper-potassium biocomplex includes the steps of: adding copper compounds to a solution containing 19 levorotatory amino acids and performing a copper-amino acid complexation reaction, then adding potassium compounds to the reaction mixture and stirring the mixture for about 3 hours at a temperature of 20 to 40 degrees Celsius. Claim 4) According to claim 1, foliar spraying of the bioactive formulation at a concentration of 6 parts per thousand is carried out 1 to 2 days before the occurrence of frost damage to increase the resistance of plants to frost damage. Claim 5) According to claim 1, the left-handed amino acids include one or more of the following: alanine, arginine, aspartic acid, cysteine, glutamic acid, valine, glycine, histidine, isoleucine, leucine, lysine, methionine, phenylalanine, proline, serine, threonine, tryptophan, tyrosine, and glutamine. Claim 6) According to claim 1, the copper ion source is used from compounds of copper nitrate, copper sulfate, copper chloride, copper acetate or copper chelates. Claim 7) According to claim 1, the potassium ion source is used from the group of potassium sulfate, potassium nitrate, potassium chloride, potassium acetate or potassium citrate. Claim 8) According to claim 1, the resulting biocomplex has high stability, complete solubility in water, rapid absorption capability, and increased bioavailability for foliar absorption. Claim 9) According to claim 1, the final product can be packaged as a stable composition without the need for filtration or phase separation.

Images