Plant extract-based superoxide dismutase bio-stress nutrient fertilizer and method of production thereof

Abstract

The application discloses a superoxide dismutase biological anti-stress nutrient fertilizer based on plant extraction and a production method thereof. The fertilizer contains, in percentage by mass, 1.0-10.0% of superoxide dismutase, 1.0-5.0% of a plant anti-stress inducing factor complex, 0.1-2.0% of organic chelated trace elements, 5.0-15.0% of a phosphorus-potassium enhancing component, 0.5-5.0% of a natural protective agent, 3.0-10.0% of a penetrating agent, 0.5-15.0% of a microbial metabolic synergist, 0.1-5.0% of a pH buffer component, 0.01-0.5% of a plant-derived preservative, and the balance of water. The superoxide dismutase is preferably derived from corn germ or spinach extract. The production method comprises the steps of enzyme activity component preparation, water phase base preparation, anti-stress function phase preparation, step-by-step mixing and primary stabilization, final product integration and secondary stabilization, and the like. Through scientific compounding and synergy of various biological active components, the application significantly enhances the resistance of crops to drought, saline-alkali, high and low temperature and the like, and promotes growth and quality improvement.

Description

Technical Field

[0001] This invention patent relates to the field of fertilizer technology, and in particular to a plant-derived superoxide dismutase bio-resistance nutrient fertilizer and its production method. Background Technology

[0002] In agricultural production, crops constantly face various abiotic stresses such as drought, salinity, high temperature, and low temperature, leading to physiological metabolic disorders, inhibited growth, and reduced yield and quality. This has become a key bottleneck restricting the sustainable development of modern agriculture. To address this challenge, developing multifunctional specialty fertilizers that integrate nutrient supply, stress resistance induction, and biostimulation is an important current research direction.

[0003] From the perspective of existing formulation design, the main shortcomings are as follows: First, the functional positioning is singular. Most products still rely on traditional macroelements or single regulatory substances (such as a certain plant hormone or amino acid), resulting in simple formulation structures that fail to activate the plant's systemic stress response network, leading to limited and non-broad-spectrum protective effects. Second, the synergy of components is poor. Existing formulations often involve simple physical mixing of raw materials with different functions, lacking scientific research on the compatibility and synergistic mechanisms between components. This fails to achieve effective connection and synergistic amplification of multiple stages such as "induction-defense-nutrition-recovery," resulting in low overall efficiency. Third, there is an over-reliance on chemically synthesized substances. Some formulations use large amounts of chemical chelating agents, synthetic preservatives, or stabilizers in pursuit of rapid effects, which may lead to problems such as soil burden, agricultural product residues, and low environmental compatibility. Finally, there is insufficient utilization of bioactive components, especially for bioenzymes such as superoxide dismutase, which have core functions in scavenging reactive oxygen species. Stabilization technologies and compound application schemes in fertilizer formulations are still immature, limiting their actual effectiveness. Summary of the Invention

[0004] In view of this, the present invention aims to provide a plant-derived superoxide dismutase bio-resistance nutrient fertilizer and its production method to solve or alleviate the technical problems existing in the prior art.

[0005] The technical solution of this invention patent embodiment is implemented as follows: a plant-derived superoxide dismutase bio-resistance nutrient fertilizer, by mass percentage, comprises the following components: superoxide dismutase 1.0%-10.0%, plant stress resistance inducing factor complex 1.0%-5.0%, organic chelated trace elements 0.1%-2.0%, phosphorus and potassium enhancing components 5.0%-15.0%, natural protective agent 0.5%-5.0%, penetrant 3.0%-10.0%, microbial metabolism synergist 0.5%-15.0%, pH buffer component 0.1%-5.0%, plant-derived preservative 0.01%-0.5%, with the balance being water.

[0006] As an improvement, the superoxide dismutase is derived from extracts of corn germ or spinach, and its enzyme activity is 3000-8000 U / g.

[0007] As an improvement, the plant stress resistance inducing factor complex comprises the following components in weight percentage: 0.5%-2.5% seaweed polysaccharide, 0.3%-1.5% betaine, 0.1%-0.7% proline analogue and 0.1%-0.3% salicylic acid.

[0008] As an improvement, the organic chelated trace elements comprise the following components by weight percentage: amino acid chelated iron 0.04%-0.8%, citric acid chelated zinc 0.03%-0.6%, humic acid chelated manganese 0.02%-0.4%, and amino acid chelated copper 0.01%-0.2%; the phosphorus and potassium reinforcing components comprise the following components by weight percentage: potassium dihydrogen phosphate 3.0%-10.0% and potassium silicate 2.0%-5.0%.

[0009] As an improvement, the natural preservative comprises the following components by weight percentage: xanthan gum 0.2%-2.0% and sodium alginate 0.3%-3.0%; the penetrant comprises the following components by weight percentage: glycerol 2.0%-6.0% and sorbitol 1.0%-4.0%; the microbial metabolic enhancer comprises the following components by weight percentage: Bacillus subtilis fermentation broth 0.3%-10.0% and Bacillus jellyoid metabolites 0.2%-5.0%; the pH buffer component is a citrate-sodium citrate buffer solution, the content of which is 0.1%-5.0% of the composition; and the plant-derived preservative is rosemary extract, the content of which is 0.01%-0.5% of the composition.

[0010] As an improvement, the composition, by weight percentage, includes the following components: superoxide dismutase 5.5%, seaweed polysaccharide 1.5%, betaine 0.9%, proline analogue 0.4%, salicylic acid 0.2%, amino acid chelated iron 0.4%, citrate chelated zinc 0.3%, humic acid chelated manganese 0.2%, amino acid chelated copper 0.1%, potassium dihydrogen phosphate 6.5%, potassium silicate 3.5%, xanthan gum 1.1%, sodium alginate 1.7%, glycerol 4.0%, sorbitol 2.5%, Bacillus subtilis fermentation broth 5.2%, gelatinous Bacillus metabolites 2.4%, citrate-sodium citrate buffer 2.5%, rosemary extract 0.25%, and the balance being water.

[0011] The production method of plant-derived superoxide dismutase bio-resistance nutrient fertilizer includes the following steps: S1. Preparation of enzyme active components: After washing and crushing the plant raw materials, the extract was extracted using a low-temperature extracting solution. After centrifugation, the supernatant was collected and concentrated and stabilized through membrane treatment to obtain a superoxide dismutase (SOD) concentrate with stable enzyme activity. S2. Preparation of aqueous base material: The natural protective agent and part of the penetrant are dissolved in water under heating and stirring to form a homogeneous colloidal solution. Then, the solution is cooled down, and the pH buffer component, plant-derived preservative and the phosphorus and potassium enhancement component are added in sequence. The solution is stirred until completely dissolved to obtain the aqueous base material. S3. Preparation of stress-resistant functional phase: The plant stress-resistant inducing factor complex and the organic chelated trace elements are mixed with the remaining penetrant in another container to form a homogeneous functional phase premix; S4. Stepwise mixing and primary stabilization: Under low-speed shear conditions, the functional phase premixed liquid obtained in step S3 is slowly added to the aqueous phase base material obtained in step S2. After mixing evenly, the microbial metabolism enhancer is added, and the mixture is continuously stirred to form a homogeneous composite emulsion. S5. Final product integration and secondary stabilization: Under temperature control and inert gas protection conditions, the SOD concentrate obtained in step S1 is slowly added to the composite emulsion obtained in step S4, homogenized using a homogenizing device, then degassed, filtered, and bottled to obtain the biological stress-resistant nutrient fertilizer.

[0012] As an improvement, in step S1, the temperature of the low-temperature extraction is 2-10°C; the membrane concentration uses an ultrafiltration membrane with a molecular weight cutoff of 5-10 kDa; and the stabilization treatment involves adding 5%-15% by mass of a permeation protectant composed of glycerol and sorbitol to the concentrate.

[0013] As an improvement, in step S2, the heating and stirring temperature is 45-55℃, and the cooling refers to cooling the solution to below 30℃; in step S4, the rotation speed of the low-speed shearing is 300-600 rpm.

[0014] As an improvement, in step S5, the temperature control refers to maintaining the material temperature below 25°C, and the pressure of the homogenization process is 15-25 MPa.

[0015] The embodiments of this invention, employing the above technical solutions, possess the following advantages: By systematically compounding core functional components such as plant-extracted superoxide dismutase (SOD), plant stress-inducing factor complexes, organically chelated trace elements, and microbial metabolic synergists, a multi-level, synergistic biological stress-resistant protection system is constructed. This formulation design effectively overcomes the technical defects of existing technologies, such as single functional components and insufficient synergistic effects, achieving the organic integration of multiple efficiencies in biostimulation, nutrient fortification, and stress defense. Specifically, exogenous plant-derived SOD synergistically activates and enhances the plant's antioxidant defense system with endogenous stress-resistance signaling substances (such as seaweed polysaccharides and betaine); organically chelated trace elements and microbial metabolites (such as Bacillus subtilis fermentation broth) produce a "bio-chemical" synergistic effect, significantly promoting root development and nutrient absorption efficiency; simultaneously, the entire formulation uses natural protectants and plant-derived preservatives, improving the product's environmental compatibility and storage stability. Ultimately, the product of this invention exhibits significant broad-spectrum stress resistance in field applications, effectively alleviating damage to crops caused by various stresses such as drought, salinity, and low temperature, while simultaneously improving yield and fruit quality, achieving a balance between stress resistance and growth promotion, resulting in significant economic benefits.

[0016] The above overview is for illustrative purposes only and is not intended to be limiting in any way. Further aspects, embodiments, and features of this invention will become readily apparent from the following detailed description, in addition to the illustrative aspects, embodiments, and features described above. Detailed Implementation

[0017] The invention will be more readily understood by referring to the following detailed description of preferred embodiments and included examples. Unless otherwise specified, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains. In case of conflict, the definitions in this specification shall prevail.

[0018] As used herein, the terms “prepared from” and “comprising” are synonymous. The terms “comprising,” “including,” “having,” “containing,” or any other variations thereof, as used herein, are intended to cover non-exclusive inclusion. For example, a composition, step, method, article, or apparatus that includes the listed elements is not necessarily limited to those elements, but may include other elements not expressly listed or elements inherent to such composition, step, method, article, or apparatus.

[0019] The conjunction "composed of..." excludes any unspecified elements, steps, or components. If used in a claim, this phrase makes the claim closed, excluding materials other than those described, except for conventional impurities associated with them. When the phrase "composed of..." appears in a clause of the body of a claim rather than immediately following it, it limits only the elements described in that clause; other elements are not excluded from the claim as a whole.

[0020] When a quantity, concentration, or other value or parameter is expressed as a range, a preferred range, or a range defined by a series of upper and lower preferred values, this should be understood as specifically disclosing all ranges formed by any pair of any upper or preferred value with any lower or preferred value, regardless of whether the range is disclosed individually. For example, when the range “1 to 5” is disclosed, the described range should be interpreted as including the ranges “1 to 4”, “1 to 3”, “1 to 2”, “1 to 2 and 4 to 5”, “1 to 3 and 5”, etc. When numerical ranges are described herein, unless otherwise stated, the range is intended to include its endpoints and all integers and fractions within that range.

[0021] The singular form includes the plural objects of discussion unless the context clearly indicates otherwise. "Optional" or "any one" means that the matter or event described thereafter may or may not occur, and the description includes both the possibility that the event occurs and the possibility that the event does not occur.

[0022] Approximate terms used in the specification and claims to modify quantities indicate that the invention is not limited to that specific quantity, but also includes acceptable modifications close to that quantity that do not alter the relevant essential function. Correspondingly, the use of "about," "approximately," etc., to modify a numerical value means that the invention is not limited to that precise value. In some instances, approximate terms may correspond to the precision of the instrument used to measure the value. In this application's specification and claims, scope definitions can be combined and / or interchanged, unless otherwise stated, these scopes include all subscopes contained therein.

[0023] Furthermore, the indefinite articles “a” and “an” preceding the elements or components of this invention do not impose any limitation on the quantity (i.e., number of times) of the elements or components. Therefore, “an” or “a” should be interpreted as including one or at least one, and the singular form of an element or component also includes the plural form, unless the quantity clearly refers to the singular form.

[0024] Furthermore, any methods and materials similar to or equivalent to those described herein may be applied to this invention. The preferred embodiments and materials described herein are for illustrative purposes only and do not limit the scope of this application.

[0025] Unless otherwise specified, the experimental methods used in the following examples are conventional methods; unless otherwise specified, the experimental materials and test strains used in the following examples were purchased from commercial channels.

[0026] Instructions on Microbial Metabolic Enhancers 1. Preparation and definition of Bacillus subtilis fermentation broth: The "Bacillus subtilis fermentation broth" of this invention is a liquid fermentation product prepared by the following standardized fermentation process.

[0027] Fermentation medium (g / L): glucose 20.0, peptone 10.0, yeast extract 5.0, NaCl 5.0, K2HPO4 2.0, MgSO4·7H2O 0.5, pH natural (approximately 7.0-7.2 before sterilization).

[0028] Fermentation process: The activated seed liquid was inoculated into the fermentation medium at an inoculation rate of 5% (v / v) and cultured at 37℃ and 180 rpm for 48 hours with shaking.

[0029] Product processing: After fermentation, the fermentation broth was centrifuged at 4°C and 8000 rpm for 15 min, and the supernatant was collected to obtain the "Bacillus subtilis fermentation broth" of this invention. The fermentation supernatant is rich in active metabolites such as lipopeptides (e.g., surfactants, iturobrine), plant growth hormones (e.g., IAA), and various enzymes produced by this strain.

[0030] 2. Preparation and definition of Bacillus jelly-like metabolites: The "Bacillus jelly-like metabolites" of this invention are prepared by the following standardized fermentation process to remove the active substances of the bacterial cells.

[0031] Fermentation medium (g / L): sucrose 10.0, Na2HPO4 2.0, MgSO4·7H2O 0.5, FeCl3 0.005, CaCO3 1.0, pH 7.0-7.5.

[0032] Fermentation process: The activated seed liquid was inoculated into the fermentation medium at an inoculation rate of 5% (v / v) and cultured at 30℃ and 150 rpm for 72 hours with shaking.

[0033] Product processing: After fermentation, the fermentation broth was centrifuged at 4°C and 10,000 rpm for 20 minutes, and the supernatant was collected, which is the "gelatinous Bacillus metabolite" described in this invention. The supernatant mainly contains bacterial extracellular polysaccharides, organic acids (such as citric acid and gluconic acid), plant hormones, and enzymes related to phosphorus solubilization and potassium solubilization.

[0034] 3. Component functions and quality control indicators: The two fermentation supernatants mentioned above serve as the core of the "microbial metabolic enhancer," and their main functions are: secreting plant growth stimulants to promote root development; producing organic acids and chelating agents to activate rhizosphere nutrients; and inhibiting soil-borne pathogens through competition and metabolic products. To ensure product consistency, the following quality control measures are implemented for the fermentation supernatants used: the surfactant titer of the Bacillus subtilis fermentation supernatant is not less than 100 mg / L (determined by high performance liquid chromatography); the polysaccharide content of the gelatinous Bacillus fermentation supernatant is not less than 2.0 g / L (determined by the phenol-sulfuric acid method). Example 1

[0035] This plant-derived superoxide dismutase (SOD) bio-nutrient fertilizer for stress resistance comprises, by weight percentage: 1.0% superoxide dismutase, 1.0% plant stress resistance inducing factor complex, 0.1% organic chelated trace elements, 5.0% phosphorus and potassium enhancing components, 0.5% natural protectant, 3.0% penetrant, 0.5% microbial metabolism synergist, 0.1% pH buffer component, 0.01% plant-derived preservative, with the balance being water.

[0036] Specifically, the superoxide dismutase is derived from extracts of corn germ or spinach, and its enzyme activity is 3000 U / g.

[0037] The plant stress resistance inducing factor complex includes seaweed polysaccharide, betaine, proline analogue and salicylic acid.

[0038] The organic chelated trace elements include amino acid chelated iron, citric acid chelated zinc, humic acid chelated manganese, and amino acid chelated copper; the phosphorus and potassium enhancing components include potassium dihydrogen phosphate and potassium silicate.

[0039] The natural preservatives include xanthan gum and sodium alginate; the penetrants include glycerin and sorbitol; the microbial metabolic enhancers include Bacillus subtilis fermentation broth and Bacillus lentiginosa metabolites; the pH buffer is a citrate-sodium citrate buffer; and the plant-derived preservative is rosemary extract.

[0040] Specifically, the plant-derived superoxide dismutase bio-resistance nutrient fertilizer, by weight percentage, includes the following components: superoxide dismutase 1.0%, seaweed polysaccharide 0.5%, betaine 0.3%, proline analogue 0.1%, salicylic acid 0.1%, amino acid chelated iron 0.04%, citrate chelated zinc 0.03%, humic acid chelated manganese 0.02%, amino acid chelated copper 0.01%, potassium dihydrogen phosphate 3.0%, potassium silicate 2.0%, xanthan gum 0.2%, sodium alginate 0.3%, glycerol 2.0%, sorbitol 1.0%, Bacillus subtilis fermentation broth 0.3%, gelatinous Bacillus metabolites 0.2%, citrate-sodium citrate buffer 0.1%, rosemary extract 0.01%, and the balance being water.

[0041] The production method of plant-derived superoxide dismutase bio-resistance nutrient fertilizer includes the following steps: S1. Preparation of enzyme active components: After washing and crushing the plant raw materials, the extract was extracted using a low-temperature extracting solution. After centrifugation, the supernatant was collected and concentrated and stabilized through membrane treatment to obtain a superoxide dismutase (SOD) concentrate with stable enzyme activity. The low-temperature extraction temperature is 2°C; the membrane concentration uses an ultrafiltration membrane with a molecular weight cutoff of 5 kDa; and the stabilization treatment involves adding 5% by mass of a permeation protectant composed of glycerol and sorbitol to the concentrate.

[0042] S2. Preparation of aqueous base material: The natural protective agent and part of the penetrant are dissolved in water under heating and stirring to form a homogeneous colloidal solution. Then, the solution is cooled down, and the pH buffer component, plant-derived preservative and the phosphorus and potassium enhancement component are added in sequence. The solution is stirred until completely dissolved to obtain the aqueous base material. The heating and stirring temperature is 45°C, and the cooling refers to cooling the solution to below 30°C.

[0043] S3. Preparation of stress-resistant functional phase: The plant stress-resistant inducing factor complex and the organic chelated trace elements are mixed with the remaining penetrant in another container to form a homogeneous functional phase premix; S4. Stepwise mixing and primary stabilization: Under low-speed shear conditions, the functional phase premixed liquid obtained in step S3 is slowly added to the aqueous phase base material obtained in step S2. After mixing evenly, the microbial metabolism enhancer is added, and the mixture is continuously stirred to form a homogeneous composite emulsion. The rotational speed of the low-speed shearing is 300 rpm.

[0044] S5. Final product integration and secondary stabilization: Under temperature control and inert gas protection conditions, the SOD concentrate obtained in step S1 is slowly added to the composite emulsion obtained in step S4, homogenized using a homogenizing device, then degassed, filtered, and bottled to obtain the biological stress-resistant nutrient fertilizer.

[0045] The temperature control refers to maintaining the material temperature below 25°C, and the homogenization process pressure is 15 MPa. Example 2

[0046] This plant-derived superoxide dismutase (SOD) bio-nutrient fertilizer for stress resistance comprises, by weight percentage: 5.5% superoxide dismutase, 3.0% plant stress resistance inducing factor complex, 1.0% organic chelated trace elements, 10.0% phosphorus and potassium enhancing components, 2.8% natural protectant, 6.5% penetrant, 7.9% microbial metabolism synergist, 2.5% pH buffer component, 0.25% plant-derived preservative, with the balance being water.

[0047] Specifically, the superoxide dismutase is derived from extracts of corn germ or spinach, and its enzyme activity is 5500 U / g.

[0048] The plant stress resistance inducing factor complex includes seaweed polysaccharide, betaine, proline analogue and salicylic acid.

[0049] The organic chelated trace elements include amino acid chelated iron, citric acid chelated zinc, humic acid chelated manganese, and amino acid chelated copper; the phosphorus and potassium enhancing components include potassium dihydrogen phosphate and potassium silicate.

[0050] The natural preservatives include xanthan gum and sodium alginate; the penetrants include glycerin and sorbitol; the microbial metabolic enhancers include Bacillus subtilis fermentation broth and Bacillus lentiginosa metabolites; the pH buffer is a citrate-sodium citrate buffer; and the plant-derived preservative is rosemary extract.

[0051] Specifically, the plant-derived superoxide dismutase bio-resistance nutrient fertilizer, by weight percentage, includes the following components: superoxide dismutase 5.5%, seaweed polysaccharide 1.5%, betaine 0.9%, proline analogue 0.4%, salicylic acid 0.2%, amino acid chelated iron 0.4%, citrate chelated zinc 0.3%, humic acid chelated manganese 0.2%, amino acid chelated copper 0.1%, potassium dihydrogen phosphate 6.5%, potassium silicate 3.5%, xanthan gum 1.1%, sodium alginate 1.7%, glycerol 4.0%, sorbitol 2.5%, Bacillus subtilis fermentation broth 5.2%, gelatinous Bacillus metabolites 2.4%, citrate-sodium citrate buffer 2.5%, rosemary extract 0.25%, and the balance being water.

[0052] The production method of plant-derived superoxide dismutase bio-resistance nutrient fertilizer includes the following steps: S1. Preparation of enzyme active components: After washing and crushing the plant raw materials, the extract was extracted using a low-temperature extracting solution. After centrifugation, the supernatant was collected and concentrated and stabilized through membrane treatment to obtain a superoxide dismutase (SOD) concentrate with stable enzyme activity. The low-temperature extraction temperature is 6°C; the membrane concentration uses an ultrafiltration membrane with a molecular weight cutoff of 8 kDa; and the stabilization treatment involves adding 10% by mass of a permeation protectant composed of glycerol and sorbitol to the concentrate.

[0053] S2. Preparation of aqueous base material: The natural protective agent and part of the penetrant are dissolved in water under heating and stirring to form a homogeneous colloidal solution. Then, the solution is cooled down, and the pH buffer component, plant-derived preservative and the phosphorus and potassium enhancement component are added in sequence. The solution is stirred until completely dissolved to obtain the aqueous base material. The heating and stirring temperature is 50°C, and the cooling refers to cooling the solution to below 30°C.

[0054] S3. Preparation of stress-resistant functional phase: The plant stress-resistant inducing factor complex and the organic chelated trace elements are mixed with the remaining penetrant in another container to form a homogeneous functional phase premix; S4. Stepwise mixing and primary stabilization: Under low-speed shear conditions, the functional phase premixed liquid obtained in step S3 is slowly added to the aqueous phase base material obtained in step S2. After mixing evenly, the microbial metabolism enhancer is added, and the mixture is continuously stirred to form a homogeneous composite emulsion. The rotational speed of the low-speed shearing is 450 rpm.

[0055] S5. Final product integration and secondary stabilization: Under temperature control and inert gas protection conditions, the SOD concentrate obtained in step S1 is slowly added to the composite emulsion obtained in step S4, homogenized using a homogenizing device, then degassed, filtered, and bottled to obtain the biological stress-resistant nutrient fertilizer.

[0056] The temperature control refers to maintaining the material temperature below 25°C, and the homogenization process pressure is 20 MPa. Example 3

[0057] This plant-derived superoxide dismutase (SOD) bio-nutrient fertilizer for stress resistance comprises, by weight percentage: 10.0% superoxide dismutase, 5.0% plant stress resistance inducing factor complex, 2.0% organic chelated trace elements, 15.0% phosphorus and potassium enhancing components, 5.0% natural protectant, 10.0% penetrant, 15.0% microbial metabolism synergist, 5.0% pH buffer component, 0.5% plant-derived preservative, with the balance being water.

[0058] Specifically, the superoxide dismutase is derived from extracts of corn germ or spinach, and its enzyme activity is 8000 U / g.

[0059] The plant stress resistance inducing factor complex includes seaweed polysaccharide, betaine, proline analogue and salicylic acid.

[0060] The organic chelated trace elements include amino acid chelated iron, citric acid chelated zinc, humic acid chelated manganese, and amino acid chelated copper; the phosphorus and potassium enhancing components include potassium dihydrogen phosphate and potassium silicate.

[0061] The natural preservatives include xanthan gum and sodium alginate; the penetrants include glycerin and sorbitol; the microbial metabolic enhancers include Bacillus subtilis fermentation broth and Bacillus lentiginosa metabolites; the pH buffer is a citrate-sodium citrate buffer; and the plant-derived preservative is rosemary extract.

[0062] Specifically, the plant-derived superoxide dismutase bio-resistance nutrient fertilizer, by weight percentage, includes the following components: superoxide dismutase 10%, seaweed polysaccharide 2.5%, betaine 1.5%, proline analogue 0.7%, salicylic acid 0.3%, amino acid chelated iron 0.8%, citrate chelated zinc 0.6%, humic acid chelated manganese 0.4%, amino acid chelated copper 0.2%, potassium dihydrogen phosphate 10.0%, potassium silicate 5.0%, xanthan gum 2.0%, sodium alginate 3.0%, glycerol 6.0%, sorbitol 4.0%, Bacillus subtilis fermentation broth 10.0%, gelatinous Bacillus metabolites 5.0%, citrate-sodium citrate buffer 5.0%, rosemary extract 0.5%, and the balance being water.

[0063] The production method of plant-derived superoxide dismutase bio-resistance nutrient fertilizer includes the following steps: S1. Preparation of enzyme active components: After washing and crushing the plant raw materials, the extract was extracted using a low-temperature extracting solution. After centrifugation, the supernatant was collected and concentrated and stabilized through membrane treatment to obtain a superoxide dismutase (SOD) concentrate with stable enzyme activity. The low-temperature extraction temperature is 10°C; the membrane concentration uses an ultrafiltration membrane with a molecular weight cutoff of 10 kDa; and the stabilization treatment involves adding 15% by mass of a permeation protectant composed of glycerol and sorbitol to the concentrate.

[0064] S2. Preparation of aqueous base material: The natural protective agent and part of the penetrant are dissolved in water under heating and stirring to form a homogeneous colloidal solution. Then, the solution is cooled down, and the pH buffer component, plant-derived preservative and the phosphorus and potassium enhancement component are added in sequence. The solution is stirred until completely dissolved to obtain the aqueous base material. The heating and stirring temperature is 55°C, and the cooling refers to cooling the solution to below 30°C.

[0065] S3. Preparation of stress-resistant functional phase: The plant stress-resistant inducing factor complex and the organic chelated trace elements are mixed with the remaining penetrant in another container to form a homogeneous functional phase premix; S4. Stepwise mixing and primary stabilization: Under low-speed shear conditions, the functional phase premixed liquid obtained in step S3 is slowly added to the aqueous phase base material obtained in step S2. After mixing evenly, the microbial metabolism enhancer is added, and the mixture is continuously stirred to form a homogeneous composite emulsion. The rotational speed of the low-speed shearing is 600 rpm.

[0066] S5. Final product integration and secondary stabilization: Under temperature control and inert gas protection conditions, the SOD concentrate obtained in step S1 is slowly added to the composite emulsion obtained in step S4, homogenized using a homogenizing device, then degassed, filtered, and bottled to obtain the biological stress-resistant nutrient fertilizer.

[0067] The temperature control refers to maintaining the material temperature below 25°C, and the homogenization process pressure is 25 MPa.

[0068] Experimental Example 1: Verification of the activity and synergistic stress resistance effect of the product's core components (indoor potted plant experiment) I. Experimental Objective: This experiment aims to simulate drought stress conditions and verify the synergistic effect between the core active component of the product of this invention—plant-derived superoxide dismutase (SOD)—and the plant stress resistance inducing factor complex. By measuring a series of physiological, biochemical, and growth recovery indicators, the effect of enhancing plant stress resistance is scientifically evaluated.

[0069] II. Materials and Methods: 1. Test materials: Plant material: Tomato variety 'Zhongza 9'. Select plump seeds, disinfect and germinate them, then sow them in seedling trays. When the seedlings have grown to four leaves and a central bud, select uniformly growing plants for transplanting.

[0070] Cultivation substrate: a mixture of peat moss, vermiculite and perlite in a ratio of 3:1:1 (v / v / v). Mix 1.5g of compound fertilizer (N-P2O5-K2O=15-15-15) into each kilogram of substrate as base fertilizer.

[0071] Experimental containers: Black plastic seedling pots (top diameter × height × bottom diameter: 12cm × 11cm × 9cm) were used uniformly. Each pot was filled with an equal amount of substrate and planted with one seedling.

[0072] Test product: CK: Deionized water.

[0073] T1 (Commercially Available Comparison): A well-known brand of amino acid water-soluble foliar fertilizer (total amino acids ≥100g / L), diluted 800 times according to the recommended concentration.

[0074] T2 (Full Formula of the Invention): The sample prepared according to Patent Example 2 is diluted 500 times with deionized water before use.

[0075] T3 (SOD-deficient group): Prepare a control standard by removing the superoxide dismutase component according to the formulation of Example 2, while keeping the other components and proportions unchanged, and dilute it 500 times before use.

[0076] T4 (Group lacking stress resistance factors): Prepare a control standard according to the formulation in Example 2, removing the plant stress resistance inducing factor complex composed of seaweed polysaccharide, betaine, proline analog, and salicylic acid, while keeping the other components and proportions unchanged, and dilute it 500 times for use.

[0077] 2. Experimental Design: The experiment was conducted in an artificial climate chamber, with the following environmental conditions set: day / night temperature 28℃ / 22℃, photoperiod 14h / 10h, and light intensity 300μmol·m⁻¹. -2 ·s -1 Relative humidity 60%-70%.

[0078] A completely randomized block design was adopted, with a total of 5 treatments, each with 15 pots (i.e., 15 replicates). 10 pots were used for destructive sampling to determine stress physiological parameters, and 5 pots were used for recovery growth observation.

[0079] All plants were allowed to recover for 7 days after transplanting before treatment began.

[0080] 3. Experimental Procedure: Step 1. Pretreatment period: After the seedling establishment period, all treatment groups were managed with normal water and fertilizer (maintaining the substrate at 70%-75% field capacity). On day 0 and day 7, the leaves were evenly sprayed with a handheld sprayer in the evening until droplets were about to drip from the leaves. The control group (CK) was sprayed with an equal amount of deionized water.

[0081] Step 2. Drought stress induction: Drought stress was initiated 48 hours after the second spraying treatment (i.e., day 9). Watering of all plants was stopped. Soil moisture was controlled by weighing, and the substrate moisture content was gradually reduced to stabilize at 40-45% of field capacity. This severe drought condition was maintained for 5 days (days 9 to 13).

[0082] Step 3. Stress index determination: On the 5th day of the drought stress maintenance period (i.e., the 13th day), 6 plants with moderate growth were randomly selected from each treatment for destructive sampling. The third fully expanded leaf from the top was selected for the determination of various physiological and biochemical indicators.

[0083] Step 4. Rehydration and Growth Recovery Observation: Immediately after the stress test sampling, the remaining 5 pots from each treatment were thoroughly rehydrated (watered until water seeped out of the drainage holes) and normal temperature, light, and water management was restored. Seven days after rehydration (i.e., day 20), plant height and the number of new leaves were measured to assess recovery capacity.

[0084] 4. Measurement Items and Methods: Superoxide dismutase (SOD) activity: The nitroblue tetrazolium (NBT) photochemical reduction inhibition method was used, and one enzyme activity unit (U) was defined as the inhibition of NBT photochemical reduction by 50%.

[0085] Peroxidase (POD) activity: The guaiacol method was used, and one unit of enzyme activity (U) was defined as a change of 0.01 in A470 per minute.

[0086] Malondialdehyde (MDA) content: determined by the thiobarbituric acid (TBA) method.

[0087] Relative water content of leaves (RWC): Measured by saturated weighing. RWC(%) = [(fresh weight - dry weight) / (saturated fresh weight - dry weight)] × 100%.

[0088] Relative chlorophyll content (SPAD value): Leaf samples at the same position were measured using a SPAD-502 chlorophyll meter.

[0089] Proline (Pro) content: extracted with sulfosalicylic acid and determined by ninhydrin colorimetric method.

[0090] Growth recovery indicators: Seven days after rehydration, plant height (from substrate surface to growth point) was measured, and the number of fully expanded new leaves was counted.

[0091] 5. Data Analysis: All data were processed using Microsoft Excel 2021. One-way ANOVA was performed using SPSS 26.0 software, and Duncan's new multiple range test was used to test for significance (p<0.05).

[0092] III. Experimental Results and Data: Table 1 Effects of different treatments under drought stress on physiological and biochemical parameters of tomato seedlings Note: Different lowercase letters after the data in the same column indicate that the difference is significant at the p<0.05 level. Data are expressed as mean ± standard deviation.

[0093] Table 2. Growth recovery of tomato seedlings under different treatments 7 days after rehydration. Note: Different lowercase letters after the data in the same column indicate that the difference is significant at the p<0.05 level.

[0094] IV. Experimental Conclusions and Summary: Based on the data in Tables 1 and 2, the following clear conclusions can be drawn: The product of this invention (T2) exhibits excellent drought protection effects: under severe drought stress, plants treated with T2 showed the highest SOD and POD activities and the lowest MDA accumulation, while maintaining the highest leaf water content, chlorophyll levels, and proline accumulation. This directly demonstrates that T2 can effectively enhance the plant's own antioxidant defense system, minimize the peroxidative damage of reactive oxygen species to the cell membrane, and thus maintain the relative stability of cell structure and photosynthetic function.

[0095] The core components exhibit significant synergistic effects: The activity of antioxidant enzymes (SOD, POD) in the T3 (SOD-deficient) group was significantly lower than that in the T2 group, while the MDA content was higher than that in the T2 group. This indicates that the exogenously added plant-derived SOD is a key initiating factor that rapidly enhances the overall antioxidant capacity of the plant and directly scavenges superoxide anions, and its role is irreplaceable.

[0096] The proline content and relative water content of the T4 group (lacking stress-resistance factors) were significantly lower than those of the T2 group, and the number of newly regenerated leaves was also slightly lower. This indicates that stress-resistance inducing factors such as seaweed polysaccharides and betaine play a core role in maintaining osmotic balance, stabilizing biofilms, and transmitting stress-resistance signals. The higher SOD activity in the T4 group may be partly due to the induction effect of SOD itself, but due to the lack of synergistic effects from stress-resistance factors, its overall stress resistance performance is still inferior to that of the full-formula T2 group.

[0097] T2's overall performance is significantly better than T3 and T4, and it is not a simple sum of the effects of the two. This confirms that the two core modules in the patent, "SOD" and "plant stress resistance inducing factor complex", have a synergistic effect of 1+1>2, which together form a multi-level and three-dimensional stress resistance protection network.

[0098] The product of this invention significantly promotes recovery growth after stress: after rehydration, plants treated with T2 showed the fastest growth rate (increased plant height and new leaf development), indicating that it helps plants recover quickly and accumulate biomass after the stress is relieved.

[0099] The product of this invention is superior to conventional commercially available products: in all measured indicators, T2 (this invention) is significantly better than T1 (commercially available amino acid foliar fertilizer), highlighting the advanced nature and unique effects of the technical path of this invention with biological enzymes and stress resistance induction as its core.

[0100] Experimental Example 2: Validation of broad-spectrum stress resistance under different stress types (growth chamber simulation experiment) I. Experimental Objective: This experiment aims to simulate two common abiotic stresses, salt stress and low temperature stress, under strictly controlled artificial climate conditions. By measuring growth, physiological and oxidative damage visualization indicators, the protective effect of the product of this invention (formulation of Example 2) against different types of stresses is systematically verified, thereby proving its broad-spectrum stress resistance.

[0101] II. Materials and Methods: 1. Test materials: Plant material: Cucumber variety 'Jinchun No. 4'. After disinfection and germination, the seeds were sown in seedling trays filled with sterilized quartz sand. When the cotyledons were fully expanded, seedlings with uniform growth were selected for hydroponics.

[0102] Hydroponic system: Use 1 / 2 strength Hoagland nutrient solution as the base culture medium, change it every 3 days, and maintain continuous aeration. Use black planting boards and 500mL opaque beakers as hydroponic containers, planting one seedling per beaker.

[0103] Test product: CK (stress control): Foliar spraying with deionized water during the stress treatment period.

[0104] T (Product treatment of this invention): Before stress treatment, the product prepared according to Example 2 and diluted 500 times was sprayed on the leaves. It was sprayed once on the 3rd day and once on the 1st day before the start of stress.

[0105] Coercion settings: A. Salt stress experiment: The experiment was conducted in an artificial climate chamber (temperature 28℃ / 22℃, photoperiod 14h / 10h, light intensity 300μmol·m⁻¹). -2 ·s -1 The stress was carried out at a humidity of 60%. When the first true leaf of the seedling was fully unfolded, the nutrient solution was replaced with 1 / 2 Hogland nutrient solution with 150mM NaCl added, and the stress was started and continued for 7 days.

[0106] B. Low-temperature stress experiment: conducted in a programmable artificial climate chamber. When the first true leaf of the seedlings was fully expanded, the ambient temperature was directly reduced from normal culture conditions (28℃ / 22℃) to low-temperature stress conditions (8℃ / 5℃, day / night), while the photoperiod and light intensity remained unchanged, for 7 days. The nutrient solution remained 1 / 2 Hoagland nutrient solution.

[0107] 2. Experimental Design: This experiment is a two-factor experiment: Factor 1 is the type of stress (salt, low temperature), and Factor 2 is the product treatment (CK, T).

[0108] For each type of stress, two treatments, CK and T, were set up, with 10 seedlings in each treatment (i.e., 10 replicates) arranged completely randomly.

[0109] All plants were pre-cultured in a hydroponic system to a uniform physiological state before being subjected to stress.

[0110] 3. Test Procedures and Measurement Items: Step 1: Pretreatment and Stress Induction After 7 days of pre-culture of seedlings, all plants in the T treatment group were foliar sprayed twice (2 days apart). The CK group was sprayed with an equal amount of water.

[0111] Twenty-four hours after the last spray, salt stress (replacing the nutrient solution with salt) and low temperature stress (lowering the ambient temperature) were initiated respectively.

[0112] Step 2: Measurement of stress indicators (day 7 of stress) Growth index determination: Six plants were randomly selected from each treatment, washed with deionized water, and blotted dry with absorbent paper. Plant height (from the base of the root to the growing point) and root length (length of the longest root) were measured. The plants were then separated into aboveground parts and root systems, blanched at 105℃ for 30 min, and dried at 75℃ to constant weight. The dry weight of the aboveground parts and the dry weight of the root system were measured separately.

[0113] Physiological and biochemical indicators were measured using the second true leaf.

[0114] Electrolyte permeability (EL): The conductivity method was used. Leaf discs were immersed in deionized water, and the initial conductivity (C1) and the final conductivity after boiling (C2) were measured. EL(%) = (C1 / C2) × 100%.

[0115] Net photosynthetic rate (Pn): The net photosynthetic rate (μmol CO2·m) of leaves was measured in situ in the growth chamber from 9:00 to 11:00 AM on the 7th day of stress using a portable photosynthesis measurement system (such as LI-6800). -2 ·s -1 The same light intensity was used in the growth chamber during the measurement.

[0116] Antioxidant enzyme activity: Superoxide dismutase (SOD): NBT photochemical reduction inhibition method.

[0117] Catalase (CAT): Activity (U / gFW) is calculated by measuring the rate of H2O2 decomposition at 240 nm using ultraviolet absorption method.

[0118] Oxidative damage histochemical staining (day 5 of stress): The second true leaf of 3 seedlings from each treatment was taken for in vivo staining for visual comparison.

[0119] Superoxide anion (O2) - •): Nitrotetrazole blue (NBT) staining was used. Leaves were immersed in 0.1% NBT (dissolved in 10 mM potassium phosphate buffer, pH 7.8) solution, vacuum permeation was followed by reaction under light for 2 hours. O2 was then applied. - The blue formazan precipitate formed during reduction indicates O2. - • Accumulation sites and intensity.

[0120] Hydrogen peroxide (H2O2): Staining was performed using 3,3'-diaminobenzidine (DAB). Leaves were immersed in a 1 mg / mL DAB solution (pH 3.8), and after vacuum permeation, the mixture was reacted under light for 8 hours. H2O2 caused DAB to polymerize under the action of peroxidase, producing a brown precipitate.

[0121] 4. Data Analysis: Data were analyzed using SPSS 26.0 software in a two-way ANOVA to determine the significant impact of adversity type, product treatment, and their interaction on various indicators. If the interaction was significant, a simple effects analysis was performed. For the same adversity type, the difference between the CK and T treatments was analyzed using an independent samples t-test (p < 0.05, p < 0.01).

[0122] III. Experimental Results and Data: Table 1. Effects of the product of the present invention on the growth and physiological indicators of cucumber seedlings under different stresses. Note: Data are expressed as mean ± standard deviation. Within the same type of adversity, ** indicates a highly significant difference between the T treatment and the CK (p < 0.01).

[0123] Table 2. Effects of the product of the present invention on the antioxidant enzyme activity of cucumber seedlings under different stresses. Note: Data are expressed as mean ± standard deviation. Within the same type of adversity, ** indicates a highly significant difference between the T treatment and the CK (p < 0.01).

[0124] Histochemical staining observation results (descriptive conclusions): NBT staining (O2) - •): Under both adverse conditions, the leaves of the CK group showed a deep blue staining, with the color being particularly deep along the veins and edges, indicating that O2 - • Significant accumulation. The leaves of the T-treated group showed a significantly lighter staining color, with both the area and intensity of the blue region being significantly reduced, indicating that the product of this invention effectively inhibited stress-induced superoxide anion bursts.

[0125] DAB staining (H2O2): Under both stress conditions, the leaves of the CK group showed obvious brown spots or patches of deposits, especially at the leaf tips and margins. The brown deposits in the leaves of the T treatment group were significantly reduced, and the color was close to light yellowish-brown, indicating that the accumulation of H2O2 was effectively controlled.

[0126] IV. Experimental Conclusions and Summary: Based on Tables 1 and 2 and the histochemical staining results, the following conclusions are drawn: The product of this invention exhibits significant protective effects against both salt and low-temperature stress, demonstrating broad-spectrum stress resistance. Under two distinctly different abiotic conditions, the T treatment, compared to the CK treatment, significantly (p<0.01) alleviated growth inhibition (decreased plant height, root length, and biomass), reduced cell membrane damage (decreased electrolyte permeability), and maintained higher photosynthetic capacity (increased net photosynthetic rate). This demonstrates that the product's protective mechanism is not targeted at a single stress factor, but rather acts on a common pathway in plant responses to stress.

[0127] The protective effect of the product is closely related to enhancing the plant's endogenous antioxidant system. Table 2 data shows that under salt and low-temperature stress, plants treated with the product of this invention exhibited significantly higher SOD and CAT activities than the corresponding controls. SOD is a key component for scavenging O2. - As the first line of defense, CAT is responsible for decomposing H2O2. The simultaneous increase in enzyme activity indicates that the product systematically enhances the plant's ability to scavenge reactive oxygen species.

[0128] The visual evidence of oxidative damage is conclusive. NBT and DAB staining results provide visual evidence: dark O2 in the leaves of the CK group. - • The precipitation of H2O2 is a direct manifestation of oxidative damage; and the significant lightening of leaf color in group T is highly consistent with the decrease in EL (membrane damage) data and the increase in antioxidant enzyme activity. This directly confirms that the product of this invention effectively curbs the explosive accumulation of reactive oxygen species (ROS) in the early stages of stress by enhancing antioxidant defense, thereby reducing the damage of oxidative damage to cell membranes and photosynthetic structures from the source.

[0129] The two-way ANOVA showed a significant interaction (p<0.05), indicating that the effectiveness of the product treatment varied under different types of adversity. However, simple effects analysis confirmed that the T treatment was significantly better than the CK treatment under each type of adversity. This suggests that the product can flexibly respond to different stresses, but the magnitude of its synergistic effect may vary depending on the nature of the stress.

[0130] Experimental Example 3: Verification of the Synergistic Effect of Microbial Metabolism Enhancers and Nutrient Absorption (Hydroculture Experiment) I. Experimental Objective This experiment aims to investigate the synergistic effect of the microbial metabolic synergist (Bacillus subtilis and Bacillus mucilaginosus metabolites) in the product of this invention, along with organically chelated trace elements and phosphorus and potassium enhancing components, on promoting crop root development and activating nutrient absorption under strictly controlled hydroponic conditions by setting up a full formula, a microbial synergist-deficient treatment, and a blank control treatment, and to verify its "biological-chemical" dual synergistic mechanism.

[0131] II. Materials and Methods: 1. Test materials: Plant material: Heading lettuce variety 'Beishen No. 3'. After sterilization and germination, the seeds were sown in petri dishes covered with moist filter paper. Once the seedlings showed signs of sprouting, they were transplanted into hydroponic sponge blocks and pre-cultured in 1 / 4 strength Hoagland nutrient solution.

[0132] Hydroponic system: A deep flow hydroponics (DFT) system is used, consisting of an opaque hydroponic tank (length × width × height: 60cm × 40cm × 20cm), a planting board, and an aeration pump. The nutrient solution is continuously aerated, with the EC value maintained at 1.2-1.5 mS / cm, and the pH value maintained at 5.8-6.2 by adjusting with dilute acid / alkali.

[0133] Test product treatment solution: Based on 1 / 2 strength Hoagland nutrient solution, the following were added respectively: CK: No products added, pure nutrient solution control.

[0134] T1 (Full Formulation Processing of the Invention): Following the product formulation of Example 2, the solution is added to the base nutrient solution at a concentration of 1000 times dilution. This ensures that the nutrient solution ultimately contains all functional components of the product except for water.

[0135] T2 (Control without microbial synergist): A control standard containing all other components (SOD, stress resistance factor, organic chelated micronutrient fertilizer, phosphorus and potassium, etc.) but without "Bacillus subtilis fermentation broth" and "gelatinous Bacillus metabolites" was added, and the concentration was converted to the same amount as T1.

[0136] 2. Experimental Design: The experiment was conducted in a smart greenhouse, with environmental conditions controlled as follows: day / night temperature 25℃ / 18℃, photoperiod 12h / 12h, and natural light supplemented to a light intensity of not less than 200 μmol·m⁻². -2 ·s -1 .

[0137] A completely randomized block design was adopted, with a total of 3 treatments (CK, T1, T2). Each treatment had 8 replicate hydroponic tanks (i.e., 8 blocks), and 4 lettuce seedlings were planted in each hydroponic tank, for a total of 32 seedlings / treatment.

[0138] When the pre-cultured lettuce seedlings have grown two true leaves, select plants with uniform growth and transplant them into the corresponding hydroponic tanks for each treatment to begin the formal experiment.

[0139] 3. Test Procedures and Measurement Items: Step 1: Handling Start-up and Culture After transplanting, 10L of fresh nutrient solution corresponding to the treatment was added to all hydroponic tanks. The nutrient solution was then changed every 5 days to maintain stable treatment components and prevent nutrient depletion.

[0140] The experiment lasted a total of 15 days.

[0141] Step 2: Sampling and Measurement (Day 15) Root morphology scanning analysis: One plantlet was randomly selected from each replicate grid for each treatment, for a total of eight plants. The plants were carefully removed, and the roots were gently rinsed with deionized water. Root images were acquired using a root scanner (e.g., Epson Expression 11000XL), and then the following parameters were analyzed using professional root analysis software (e.g., WinRHIZO): Total root length; Total root surface area; Total number of root tips.

[0142] Nutrient content determination in plants: Scanned plants were separated into aboveground parts and roots. After blanching at 105℃ for 30 minutes, they were dried at 75℃ to constant weight, and the dry weight was measured. The dried samples were ground and analyzed using conventional methods. Macroelements: The total nitrogen (N), total phosphorus (P), and total potassium (K) content in the aboveground parts were determined by sulfuric acid-hydrogen peroxide digestion and flow analyzer.

[0143] Trace elements: The contents of iron (Fe), zinc (Zn), manganese (Mn), and copper (Cu) in the aboveground parts were determined by nitric acid-perchloric acid digestion and inductively coupled plasma mass spectrometry (ICP-MS).

[0144] Root activity assay: Three additional plants from each treatment that had not undergone destructive scanning were collected, and fresh root samples were taken. Root dehydrogenase activity was determined using the triphenyltetrazolium chloride (TTC) reduction method. The activity was expressed as TTC reducing strength (μg·g⁻¹). -1 ·h -1 () indicates root vitality.

[0145] 4. Data Analysis: All data were analyzed using SPSS 26.0 software for one-way ANOVA, and multiple comparisons were performed using Duncan's new multiple range method. The significance level was set at p < 0.05.

[0146] III. Experimental Results and Data: Table 1 Effects of different treatments on root morphology and vigor of lettuce Note: Different lowercase letters after the data in the same column indicate that the difference is significant at the p<0.05 level. Data are expressed as mean ± standard deviation.

[0147] Table 2. Effects of different treatments on dry matter accumulation and nutrient content in the aboveground parts of lettuce. Note: Different lowercase letters after the data in the same column indicate significant differences at the p<0.05 level. Data are expressed as mean ± standard deviation. DW: Dry weight.

[0148] IV. Experimental Conclusions and Summary: Based on the data in Tables 1 and 2, the following clear conclusions can be drawn: The complete formula (T1) of this invention has significant advantages in promoting root development and nutrient absorption. The total root length, surface area, number of root tips, and root activity of lettuce treated with T1 were all significantly higher than those treated with CK and T2. The well-developed root architecture and higher physiological activity laid a solid foundation for efficient nutrient absorption, which was directly reflected in the highest accumulation of aboveground dry weight and most nutrient contents (N, P, K, Fe, Zn, Cu) in the T1 treatment.

[0149] Microbial metabolic synergists are a key driver of the "synergistic effect." Although the T2 (microbial synergist-deficient) treatment contained organic chelated micronutrients and phosphorus and potassium components, it significantly outperformed the pure nutrient solution control (CK) in all indicators, demonstrating the effectiveness of the chemical nutrient components in the formulation. However, T2 was significantly lower than T1 (complete formulation) in all indicators. This indicates that: Metabolites of Bacillus subtilis and Bacillus mucilaginosa (such as plant growth hormone IAA, cytokinins, organic acids, polysaccharides, etc.) significantly promoted root morphogenesis and physiological activity (data in Table 1), which T2 could not achieve.

[0150] These metabolites exhibit strong positive interactions with organically chelated trace elements (especially Fe and Zn). For Fe and Zn absorption, T1 increased by approximately 28% and 36% compared to T2, respectively, far exceeding the increases for macroelements such as N, P, and K. This may be because microbial metabolites further activate the rhizosphere microenvironment, secreted chelating agents or protons promote the dissolution of solid nutrients, or by regulating the expression of genes related to iron and zinc absorption and transport within the plant, thereby amplifying the absorption efficiency of organically chelated nutrients.

[0151] This demonstrates the scientific validity of the "bio-chemical" synergistic effect model. The experimental results validate the logic of the formulation design of this invention: microbial metabolites, as "biostimulants," primarily target root development and activation, while organic chelated nutrients, as "high-efficiency nutrient sources," provide directly absorbable forms. The combination of the two produces a significant synergistic effect in root construction and nutrient absorption (especially trace elements), while the chemical component (T2) alone cannot achieve the same effect.

[0152] Experiment Example 4: Evaluation of the comprehensive application effect and economic benefits in the field (field experiment) I. Experimental Objective: This experiment aims to comprehensively evaluate the protective effect of the product of this invention under natural adverse conditions (high temperature) in actual field production, using high-value-added fruit trees such as grapes, as well as its comprehensive impact on fruit yield, quality and tree health. Based on the input-output ratio, an economic benefit analysis will be conducted to verify its value and promotion potential in actual agricultural applications.

[0153] II. Materials and Methods: 1. Overview of the test site: Location: A standardized vineyard in Binchuan County, Dali Prefecture, Yunnan Province (25.8°N, 100.6°E).

[0154] Soil: Sandy loam, pH 7.2, organic matter content 1.5%, available nitrogen 85 mg / kg, available phosphorus (P2O5) 35 mg / kg, available potassium (K2O) 180 mg / kg.

[0155] The test crop was grape (Vitis vinifera L.), variety 'Sunshine Rose', four-year-old, with a plant spacing of 2.5m × 3.0m, and cultivated under a "T"-shaped trellis.

[0156] Climate characteristics: In the experimental year (2023), during the grape berry expansion period (late May to mid-June), there was an abnormally high temperature weather with a daily maximum temperature of ≥35℃ for more than a week, which constituted a natural high temperature stress condition.

[0157] 2. Experimental Design: Processing settings: CK (Common Management Control): Fertilizer and water management, pest and disease control, and flower and fruit management were carried out according to the park's standardized plan. For foliar fertilizer, commercially available potassium dihydrogen phosphate (500 times dilution) was sprayed only once after flowering and at the beginning of color change.

[0158] T (Treatment of the product of this invention): Based on the same routine management as CK, apply the product of this invention during the following four key phenological periods (Example 2, diluted 600 times for foliar spraying): 1. Budding and leaf unfolding period (around March 25th) 2. Before flowering (around April 20th) 3. Young fruit enlargement stage (around May 15th, before the arrival of high temperatures) 4. Early stage of color change (around June 25th) Experimental Design: A randomized block design was adopted. Thirty consecutive grapevines with similar growth vigor and flowering quantity were selected as one plot, with four replicates per treatment, for a total of eight plots and 240 grapevines. Three guard rows were set between plots.

[0159] 3. Field management: Apart from foliar treatment, all the soil fertilization (mainly organic fertilizer as base fertilizer and water-soluble fertilizer as top dressing), irrigation, pest and disease control (mainly preventing downy mildew, powdery mildew and thrips), and flower and fruit management (flower and fruit thinning, seedless fruit preservation treatment, bagging) measures in all communities are completely consistent and are operated by the same technical team.

[0160] Spraying is carried out after 16:00 on sunny afternoons, using a backpack electric sprayer to evenly spray the leaves, the back of the leaves and the fruit clusters, until they are wet but not dripping.

[0161] 4. Measurement Items and Methods: Adversity response indicators: Sunburn rate: One week after the end of the high-temperature weather (June 20), 10 bunches of grapes were randomly surveyed in each plot, and the number of fruits with obvious scalding, dents, and browning on the fruit surface was counted. Sunburn rate (%) = (Number of sunburned fruits / Total number of surveyed fruits) × 100%.

[0162] Yield and marketability indicators (measured uniformly at the fruit's commercial maturity stage, August 10th): Single ear weight: 20 representative ears of fruit were randomly selected from each plot, weighed using an electronic scale, and the average weight (g) was taken.

[0163] Single fruit weight: 50 fruits were randomly selected from the above-mentioned bunches and weighed, and the average value (g) was calculated.

[0164] Soluble solids (sugar content): The sugar content of the juice of 20 fruits in each plot was measured using a digital refractometer, and the average value (°Brix) was taken.

[0165] Fruit firmness: Using a fruit firmness tester, the firmness of the peeled portion of the center of the sunny side of 20 fruits in each plot was measured, and the average value (kg / cm²) was taken. 2 ).

[0166] Fruit surface color uniformity: scored by 3 experienced technicians in a blind evaluation (1-5 points, 1 point is extremely uneven, 5 points is uniform and bright green).

[0167] Commercial fruit rate: The percentage (%) of the total number of clusters harvested in each plot that meet the first-grade commercial fruit standard for this variety (single cluster weight 500-800g, uniform fruit size, no sunburn, no disease or insect spots, and good color).

[0168] Tree health indicators (measured one week after harvest, on August 17): SPAD value of functional leaves: The SPAD value of the 3rd to 5th leaves from the bottom on the corresponding branches of 10 fruit clusters in each plot was measured using a chlorophyll meter, and the average value was taken.

[0169] Downy mildew disease index: Investigate the lower leaves of all plants in each plot and classify them according to the proportion of diseased area to leaf area (0: no disease; 1: ≤5%; 3: 6%-25%; 5: 26%-50%; 7: >50%), and calculate the disease index.

[0170] 5. Economic Benefit Analysis: Record the differences in input costs for each treatment (mainly product purchase and manual spraying costs).

[0171] The average yield per acre of each treatment plot was calculated based on the purchase price of different grades of 'Sunshine Rose' grapes by local buyers.

[0172] Calculate the output-input ratio.

[0173] 6. Data Analysis: Independent samples t-tests were performed using SPSS 26.0 software to compare the significance of differences in each indicator between the CK and T treatments (p<0.05, p<0.01).

[0174] III. Experimental Results and Data: Table 1. Effects of the product of this invention on the stress response, yield, and fruit quality of 'Sunshine Rose' grapes. Note: Data are expressed as mean ± standard deviation. ** indicates a significant difference (p<0.05), and ** indicates an extremely significant difference (p<0.01).

[0175] Table 2. Economic Benefit Analysis of Applying the Product of This Invention to 'Sunshine Rose' Grapes (per acre) IV. Experimental Conclusions and Summary: Based on the data in Tables 1 and 2, the following clear conclusions can be drawn: This invention significantly enhances the resistance of grapes to natural high-temperature stress. Under abnormally high temperatures, the sunburn rate in treatment T (3.2%) was significantly lower than that in treatment CK (8.7%), with a relative reduction of over 60%. This directly demonstrates that the product can effectively activate the plant's stress-resistance physiological mechanisms (such as enhancing cell membrane stability and improving antioxidant capacity) under field conditions, reducing the direct physical damage to young fruits caused by sudden extreme environments and ensuring a good yield.

[0176] This invention comprehensively improves both fruit yield and core quality. The T treatment not only increases potential yield by reducing sunburn loss, but also promotes uniform fruit enlargement by enhancing tree nutrition and stress resistance, resulting in a significant increase in average bunch weight and individual fruit weight. More importantly, the sugar content (19.8°Brix vs. 18.2°Brix), firmness, and color uniformity of the T-treated fruit are significantly improved, ultimately leading to a substantial increase in the marketable fruit rate from 78.5% to 89.3%. This demonstrates that the product not only maintains yield, but its core function lies in improving quality, and quality is the decisive factor in the economic benefits of high-end fruits such as 'Sunshine Rose'.

[0177] This invention enhances the health of trees in the later stages, demonstrating its biological health-preserving function. After harvesting, the functional leaves of T-treated plants showed higher chlorophyll content (SPAD value) and lower downy mildew disease index. This indicates that through repeated application in the early stages, the product systematically strengthens the tree vigor, improves the photosynthetic efficiency and disease resistance potential of the leaves, and is beneficial for post-harvest nutrient accumulation and safe overwintering, laying the foundation for a bumper and stable yield in the following year.

[0178] The product of this invention has extremely high economic benefits and application value. Economic benefit analysis (Table 2) shows that although treatment T increased input costs by approximately 180 yuan per mu, the total output value per mu increased by 3435 yuan due to the significant improvement in the marketable fruit rate (especially the proportion of first-grade fruit) and sugar content, resulting in a net increase in income of 3255 yuan, and a production-input ratio as high as 18.1:1. This eloquently demonstrates from an economic perspective that using the product of this invention is a high-return production input that can truly create significant value for growers.

[0179] The above description is merely a specific embodiment of the present invention, but the scope of protection of the present invention is not limited thereto. Any person skilled in the art can easily conceive of various variations or substitutions within the technical scope disclosed in the present invention, and these should all be included within the scope of protection of the present invention. Therefore, the scope of protection of the present invention should be determined by the scope of the claims.

Claims

1. A plant-derived superoxide dismutase bio-stress-resistant nutrient fertilizer, characterized in that, By weight percentage, it includes the following components: superoxide dismutase 1.0%-10.0%, plant stress resistance inducing factor complex 1.0%-5.0%, organic chelated trace elements 0.1%-2.0%, phosphorus and potassium enhancing components 5.0%-15.0%, natural protective agent 0.5%-5.0%, penetrant 3.0%-10.0%, microbial metabolism synergist 0.5%-15.0%, pH buffer component 0.1%-5.0%, plant-derived preservative 0.01%-0.5%, with the balance being water.

2. The superoxide dismutase-based bio-stress-resistant nutrient fertilizer based on plant extraction according to claim 1, characterized in that, The superoxide dismutase is derived from extracts of corn germ or spinach, and its enzyme activity is 3000-8000 U / g.

3. The superoxide dismutase-based bio-stress-resistant nutrient fertilizer based on plant extracts according to claim 1, characterized in that, The plant stress resistance inducing factor complex comprises the following components in weight percentage: 0.5%-2.5% seaweed polysaccharide, 0.3%-1.5% betaine, 0.1%-0.7% proline analogue and 0.1%-0.3% salicylic acid.

4. The superoxide dismutase-based bio-stress-resistant nutrient fertilizer based on plant extraction according to claim 1, characterized in that, The organic chelated trace elements comprise the following components by weight percentage: amino acid chelated iron 0.04%-0.8%, citric acid chelated zinc 0.03%-0.6%, humic acid chelated manganese 0.02%-0.4%, and amino acid chelated copper 0.01%-0.2%; the phosphorus and potassium reinforcing components comprise the following components by weight percentage: potassium dihydrogen phosphate 3.0%-10.0% and potassium silicate 2.0%-5.0%.

5. The superoxide dismutase-based bio-stress-resistant nutrient fertilizer based on plant extraction according to claim 1, characterized in that, The natural preservative comprises the following components by weight percentage: xanthan gum 0.2%-2.0% and sodium alginate 0.3%-3.0%; the penetrant comprises the following components by weight percentage: glycerol 2.0%-6.0% and sorbitol 1.0%-4.0%; the microbial metabolic enhancer comprises the following components by weight percentage: Bacillus subtilis fermentation broth 0.3%-10.0% and Bacillus jellyoid metabolites 0.2%-5.0%; the pH buffer is a citrate-sodium citrate buffer solution, with a content of 0.1%-5.0% of the composition; the plant-derived preservative is rosemary extract, with a content of 0.01%-0.5% of the composition.

6. The superoxide dismutase bio-resistance nutrient fertilizer based on plant extracts according to any one of claims 1-5, characterized in that, The composition, by weight percentage, includes the following: superoxide dismutase 5.5%, seaweed polysaccharide 1.5%, betaine 0.9%, proline analogue 0.4%, salicylic acid 0.2%, amino acid chelated iron 0.4%, citrate chelated zinc 0.3%, humic acid chelated manganese 0.2%, amino acid chelated copper 0.1%, potassium dihydrogen phosphate 6.5%, potassium silicate 3.5%, xanthan gum 1.1%, sodium alginate 1.7%, glycerol 4.0%, sorbitol 2.5%, Bacillus subtilis fermentation broth 5.2%, gelatinous Bacillus metabolites 2.4%, citrate-sodium citrate buffer 2.5%, rosemary extract 0.25%, and the balance being water.

7. A method for producing a plant-extracted superoxide dismutase bio-resistance nutrient fertilizer as described in any one of claims 1-6, characterized in that, Includes the following steps: S1. Preparation of enzyme active components: After washing and crushing the plant raw materials, the extract was extracted using a low-temperature extracting solution. After centrifugation, the supernatant was collected and concentrated and stabilized through membrane treatment to obtain a superoxide dismutase (SOD) concentrate with stable enzyme activity. S2. Preparation of aqueous base material: The natural protective agent and part of the penetrant are dissolved in water under heating and stirring to form a homogeneous colloidal solution. Then, the solution is cooled down, and the pH buffer component, plant-derived preservative and the phosphorus and potassium enhancement component are added in sequence. The solution is stirred until completely dissolved to obtain the aqueous base material. S3. Preparation of stress-resistant functional phase: The plant stress-resistant inducing factor complex and the organic chelated trace elements are mixed with the remaining penetrant in another container to form a homogeneous functional phase premix; S4. Stepwise mixing and primary stabilization: Under low-speed shear conditions, the functional phase premixed liquid obtained in step S3 is slowly added to the aqueous phase base material obtained in step S2. After mixing evenly, the microbial metabolism enhancer is added, and the mixture is continuously stirred to form a homogeneous composite emulsion. S5. Final product integration and secondary stabilization: Under temperature control and inert gas protection conditions, the SOD concentrate obtained in step S1 is slowly added to the composite emulsion obtained in step S4, homogenized using a homogenizing device, then degassed, filtered, and bottled to obtain the biological stress-resistant nutrient fertilizer.

8. The production method of the plant-extracted superoxide dismutase bio-stress-resistant nutrient fertilizer according to claim 7, characterized in that: In step S1, the low-temperature extraction temperature is 2-10℃; the membrane concentration uses an ultrafiltration membrane with a molecular weight cutoff of 5-10kDa; the stabilization treatment involves adding 5%-15% by mass of a permeation protectant composed of glycerol and sorbitol to the concentrate.

9. The production method of the plant-extracted superoxide dismutase bio-stress-resistant nutrient fertilizer according to claim 7, characterized in that: In step S2, the heating and stirring temperature is 45-55℃, and the cooling refers to cooling the solution to below 30℃; in step S4, the rotation speed of the low-speed shearing is 300-600 rpm.

10. The production method of the plant-extracted superoxide dismutase bio-stress-resistant nutrient fertilizer according to claim 7, characterized in that, In step S5, temperature control refers to maintaining the material temperature below 25°C, and the pressure of homogenization treatment is 15-25 MPa.

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