Nutritional agent for promoting disease resistance of pine trees and preparation method of nutritional agent

Abstract

The invention relates to the technical field of pine tree disease-resistant nutritional agents, in particular to a pine tree disease-resistant nutritional agent and a preparation method of the pine tree disease-resistant nutritional agent. 8-12 parts of a growth regulator compound; 20-30 parts of an amino acid compound; 10 to 15 parts of a polysaccharide compound; 5-8 parts of an antioxidant; 2-4 parts of a surfactant; 3-5 parts of a buffering agent; according to the invention, the trace elements, the growth regulator, the amino acid and the polysaccharide are uniquely and optimally combined. From the aspect of molecular level, complex synergistic effects exist among the components. For example, microelements such as zinc (Zn) and manganese (Mn) are taken as cofactors of various enzyme systems and can be combined with specific functional groups (such as sulfydryl-SH and carboxyl-COOH) in protein molecules to form a metalloenzyme compound, so that key physiological processes such as photosynthesis and respiration are catalyzed.

Description

Technical Field

[0001] The invention relates to the technical field of pine disease resistance nutrients, in particular to a pine disease resistance promoting nutrient and a preparation method thereof. Background Art

[0002] As an important ecological and economic tree species, pine trees play a key role in the global forest ecosystem. However, in recent years, the rampant diseases such as pine wood nematode disease have seriously threatened the survival and growth of pine trees. Traditional prevention and control methods mainly rely on chemical pesticides, which not only have limited effects, but may also have a negative impact on the environment. Therefore, the development of an environmentally friendly nutrient that can both promote the growth of pine trees and enhance their disease resistance has become a hot topic in current forestry research.

[0003] In the prior art, there are some nutritional formulas for pine trees, but most of them have the following shortcomings: first, most formulas only focus on the supplementation of a single or limited number of nutrients, ignoring the integrity and complexity of plant nutritional needs; second, many formulas lack the mechanism to activate the plant's own immune system and cannot fundamentally improve the plant's disease resistance; third, existing formulas often ignore the bioavailability of nutrients in the soil, resulting in poor actual application effects; finally, most formulas do not take environmental friendliness into consideration and may cause potential harm to the soil ecosystem. Summary of the invention

[0004] The present invention aims to solve the above technical problems and provide a multi-synergistic, resistance-inducing, highly absorbable and environmentally friendly pine disease-resistant nutrient and a preparation method thereof.

[0005] The object of the present invention is to provide a nutrient for promoting pine disease resistance, which comprises the following components by weight:

[0006] 15-25 parts of trace element complex;

[0007] 8-12 parts of growth regulator complex;

[0008] Amino acid complex 20-30 parts;

[0009] 10-15 parts of polysaccharide complex;

[0010] 5-8 parts of antioxidants;

[0011] 2-4 parts of surfactant;

[0012] 3-5 parts of buffer;

[0013] The carrier balance is up to 100 parts.

[0014] Preferably, the trace element complex comprises:

[0015] 3-5 parts of zinc citrate;

[0016] 3-5 parts of manganese citrate;

[0017] Chelated iron 5-8 parts;

[0018] 2-4 parts of copper gluconate;

[0019] 2-3 parts of borax.

[0020] Preferably, the growth regulator complex comprises: 2-3 parts of indole-3-acetic acid;

[0021] 2-3 parts of 6-benzylaminopurine;

[0022] Gibberellic acid A32-3 copies;

[0023] 2-3 parts of salicylic acid.

[0024] Preferably, the amino acid complex comprises: 5-8 parts of L-arginine;

[0025] 5-8 parts of L-glycine;

[0026] L-glutamic acid 5-7 parts;

[0027] L-proline 5-7 parts.

[0028] Preferably, the polysaccharide complex comprises:

[0029] 5-8 parts of chitosan with a molecular weight of 100-300 kDa; 5-7 parts of β-1,3-glucan with a molecular weight of 50-150 kDa. Preferably, the antioxidant comprises:

[0030] 3-5 parts of ascorbic acid;

[0031] 2-3 parts of α-tocopherol.

[0032] Preferably, the surfactant is polysorbate 80; the buffer comprises 1.5-2.5 parts of sodium citrate and 1.5-2.5 parts of potassium dihydrogen phosphate; and the carrier is deionized water.

[0033] A method for preparing the nutrient for promoting pine disease resistance comprises the following steps:

[0034] (1) preparing a trace element complex solution;

[0035] (2) preparing a growth regulator complex solution;

[0036] (3) preparing an amino acid complex solution;

[0037] (4) preparing a polysaccharide complex solution;

[0038] (5) preparing an antioxidant solution;

[0039] (6) mixing the solutions obtained in steps (1) to (5), and adding a surfactant, a buffer and a carrier to obtain a main agent;

[0040] (7) homogenizing the main agent;

[0041] (8) sterilizing the homogenized solution;

[0042] (9) Fill the sterilized solution.

[0043] Preferably, in step (1), the components of the trace element complex are sequentially added into deionized water at 50-60° C., with a stirring speed of 200-300 rpm and a dissolution time of 30-40 minutes, and then the pH is adjusted to 5.5-6.5.

[0044] As a preference, the method further comprises the following steps:

[0045] In the step (7), a high pressure homogenizer is used for treatment at a pressure of 50-60 MPa, and the mixture is circulated for 3-5 times;

[0046] In the step (8), a membrane filtration sterilization method is used, and filtration is performed using a 0.22 μm filter membrane;

[0047] In the step (9), the sterilized solution is filled into a pre-sterilized container under aseptic conditions and sealed.

[0048] The innovative features and technical effects of the present invention are mainly reflected in the following aspects:

[0049] 1. Multi-component synergistic formula: The present invention uniquely optimizes the combination of trace elements, growth regulators, amino acids and polysaccharides. From a molecular level, there is a complex synergistic effect between these components. For example, trace elements such as zinc (Zn) and manganese (Mn), as cofactors of various enzyme systems, can bind to specific functional groups in protein molecules (such as thiol-SH, carboxyl-COOH) to form metal enzyme complexes, thereby catalyzing key physiological processes such as photosynthesis and respiration. At the same time, growth regulators such as indole-3-acetic acid (IAA) can be recognized by specific receptors through the conjugated system of its indole ring, triggering a series of signal transduction processes, and ultimately regulating gene expression and cell physiological activities. This multi-component synergistic effect significantly improves the overall effect of the nutrient, far exceeding the simple superposition of a single ingredient.

[0050] 2. Inducing systemic resistance: The present invention cleverly introduces salicylic acid as an inducer, which can activate the plant's systemic acquired resistance (SAR). Salicylic acid molecules bind to specific receptor proteins, triggering a series of signal transductions, ultimately leading to enhanced expression of defense-related genes. This molecular-level regulatory mechanism enables pine trees to respond to pathogen attacks faster and more effectively, greatly improving their disease resistance.

[0051] 3. Innovation in sustained-release technology: The present invention uses chitosan and β-1,3-glucan as polysaccharide complexes, which not only serve as a nutrient source, but also form a unique microcapsule structure. This structure is based on intermolecular hydrogen bonds and electrostatic interactions, and can slowly release other active ingredients and extend the shelf life of the nutrient. This innovation greatly improves nutrient utilization efficiency and reduces the frequency of application.

[0052] 4. Biocompatibility design: The present invention selects citrate and gluconate as carriers of trace elements. These organic acid radicals can form stable chelates with metal ions. This design not only improves the bioavailability of elements, but also reduces the potential negative impact on soil ecosystems. From a chemical point of view, the formation of these chelates involves coordination chemistry. Metal ions form coordination bonds with the lone pair of electrons of organic acid radicals through their empty orbitals, which are both stable and easily absorbed by plants.

[0053] 5. Antioxidant protection system: Ascorbic acid and α-tocopherol added in the present invention form a synergistic antioxidant network. As a water-soluble antioxidant, ascorbic acid can remove free radicals in the aqueous phase; while α-tocopherol, as a fat-soluble antioxidant, mainly protects cell membranes from damage by lipid peroxidation. The synergistic effect of these two substances not only protects other active ingredients in the formula, but also directly enhances the antioxidant capacity of pine.

[0054] 6. Optimization of pH buffer system: The present invention constructs an efficient pH buffer system by combining sodium citrate and potassium dihydrogen phosphate. This system is based on the principle of polybasic weak acid-weak base balance and can remain stable in a wide pH range, ensuring the stability and bioavailability of nutrient elements under different soil conditions.

[0055] 7. Preparation process innovation: The present invention adopts high-pressure homogenization technology, which significantly improves the stability and uniformity of the nutrient through shear force and cavitation. This process not only ensures the full mixing of the components, but also can change the spatial configuration of certain macromolecules through mechanical force, further optimizing their biological activity.

[0056] The beneficial effects of the present invention are not only reflected in significantly improving the growth rate and disease resistance of pine trees, but also in improving the soil microbial environment, improving nutrient utilization efficiency, reducing environmental burden, etc. It is particularly worth mentioning that the present invention indirectly promotes plant growth and disease resistance by optimizing the rhizosphere microbial community, and this effect is rarely reported in the prior art.

[0057] In summary, the present invention provides a new idea and method for the development of pine disease-resistant nutrients through the cross-integration of multidisciplinary knowledge and innovative technical design. It not only solves many problems existing in the prior art, but also achieves technical breakthroughs in many aspects, and has important theoretical significance and application value. DETAILED DESCRIPTION

[0058] In order to make the purpose, technical solution and advantages of the present invention clearer, the technical solution of the present invention will be clearly and completely described below in conjunction with the specific embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without making creative work are within the scope of protection of the present invention.

[0059] Example 1: A nutrient for promoting pine disease resistance and its preparation method

[0060] The present embodiment provides a nutrient for promoting disease resistance of pine trees, which includes the following components in parts by weight: 15 parts of trace element complex, 8 parts of growth regulator complex, 20 parts of amino acid complex, 10 parts of polysaccharide complex, 5 parts of antioxidant, 2 parts of surfactant, 3 parts of buffer, and deionized water as a carrier, with the remainder being up to 100 parts.

[0061] Among them, the trace element complex includes 3 parts of zinc citrate, 3 parts of manganese citrate, 5 parts of chelated iron, 2 parts of copper gluconate and 2 parts of borax. The growth regulator complex includes 2 parts of indole-3-acetic acid, 2 parts of 6-benzylaminopurine, 32 parts of gibberellin A and 2 parts of salicylic acid. The amino acid complex includes 5 parts of L-arginine, 5 parts of L-glycine, 5 parts of L-glutamic acid and 5 parts of L-proline. The polysaccharide complex includes 5 parts of chitosan with a molecular weight of 100kDa and 5 parts of β-1,3-glucan with a molecular weight of 50kDa. The antioxidant includes 3 parts of ascorbic acid and 2 parts of α-tocopherol. The surfactant is polysorbate 80. The buffer includes 1.5 parts of sodium citrate and 1.5 parts of potassium dihydrogen phosphate.

[0062] The preparation method of the nutrient for promoting pine disease resistance in this embodiment comprises the following steps:

[0063] (1) Preparation of trace element complex solution: Each trace element component was added into deionized water at 50° C. in sequence, with a stirring speed of 200 rpm and a dissolution time of 30 minutes. Subsequently, the pH was adjusted to 5.5 using 0.1 mol / L sodium hydroxide solution.

[0064] (2) Preparation of growth regulator complex solution: First, indole-3-acetic acid was dissolved in 10 mL of ethanol. Second, 6-benzylaminopurine was dissolved in 10 mL of 1 mol / L sodium hydroxide solution. Then, gibberellin A3 was dissolved in 10 mL of ethanol. Finally, salicylic acid was dissolved in 10 mL of deionized water and stirred at 50°C until completely dissolved. The above four solutions were mixed, diluted to 100 mL with deionized water, and stirred at room temperature for 15 minutes.

[0065] (3) Preparation of amino acid complex solution: Each amino acid component was added into deionized water at 40° C., with a stirring speed of 150 rpm and a dissolution time of 20 minutes.

[0066] (4) Preparation of polysaccharide complex solution: First, chitosan was added to 1% (v / v) acetic acid solution and stirred at 40°C for 2 hours until completely dissolved. Second, β-1,3-glucan was added to deionized water and stirred at 60°C for 1 hour until completely dissolved. Finally, the above two solutions were mixed and stirred at room temperature for 30 minutes.

[0067] (5) Preparation of antioxidant solution: Add ascorbic acid and α-tocopherol into deionized water at 30° C. and stir to dissolve in dark conditions for 15 minutes.

[0068] (6) Preparation of the main agent: The solutions prepared in steps (1) to (5) were added to the batching tank in sequence, with a stirring speed of 300 rpm and a temperature of 25° C. Polysorbate 80 was added and stirred for 10 minutes until uniform. A buffer was then added and the pH was adjusted to 6.0. Deionized water was finally added to 95% of the total weight and stirred for 30 minutes.

[0069] (7) Homogenization treatment: The main agent is treated by a high-pressure homogenizer at a pressure of 50 MPa and cycled 3 times.

[0070] (8) Sterilization: Use membrane filtration sterilization method, use 0.22 μm filter membrane, and filter under sterile conditions.

[0071] (9) Filling: Fill the sterilized solution into a pre-sterilized container under aseptic conditions and seal it.

[0072] Example 2: A nutrient for promoting pine disease resistance and its preparation method

[0073] The present embodiment provides a nutrient for promoting disease resistance of pine trees, which includes the following components in parts by weight: 20 parts of trace element complex, 10 parts of growth regulator complex, 25 parts of amino acid complex, 12.5 parts of polysaccharide complex, 6.5 parts of antioxidant, 3 parts of surfactant, 4 parts of buffer, and deionized water as a carrier, with the remainder being up to 100 parts.

[0074] Among them, the trace element complex includes 4 parts of zinc citrate, 4 parts of manganese citrate, 6.5 parts of chelated iron, 3 parts of copper gluconate and 2.5 parts of borax. The growth regulator complex includes 2.5 parts of indole-3-acetic acid, 2.5 parts of 6-benzylaminopurine, 2.5 parts of gibberellin A3 and 2.5 parts of salicylic acid. The amino acid complex includes 6.5 parts of L-arginine, 6.5 parts of L-glycine, 6 parts of L-glutamic acid and 6 parts of L-proline. The polysaccharide complex includes 6.5 parts of chitosan with a molecular weight of 200kDa and 6 parts of β-1,3-glucan with a molecular weight of 100kDa. The antioxidant includes 4 parts of ascorbic acid and 2.5 parts of α-tocopherol. The surfactant is polysorbate 80. The buffer includes 2 parts of sodium citrate and 2 parts of potassium dihydrogen phosphate.

[0075] The preparation method of the nutrient for promoting pine disease resistance in this embodiment comprises the following steps:

[0076] (1) Preparation of trace element complex solution: Each trace element component was added into deionized water at 55° C. in sequence, with a stirring speed of 250 rpm and a dissolution time of 35 minutes. Subsequently, the pH was adjusted to 6.0 using 0.1 mol / L sodium hydroxide solution.

[0077] (2) Preparation of growth regulator complex solution: First, indole-3-acetic acid was dissolved in 12.5 mL of ethanol. Second, 6-benzylaminopurine was dissolved in 12.5 mL of 1 mol / L sodium hydroxide solution. Then, gibberellin A3 was dissolved in 12.5 mL of ethanol. Finally, salicylic acid was dissolved in 12.5 mL of deionized water and stirred at 55°C until completely dissolved. The above four solutions were mixed, diluted to 100 mL with deionized water, and stirred at room temperature for 17.5 minutes.

[0078] (3) Preparation of amino acid complex solution: Each amino acid component was added into deionized water at 45° C., with a stirring speed of 175 rpm and a dissolution time of 25 minutes.

[0079] (4) Preparation of polysaccharide complex solution: First, chitosan was added to 1% (v / v) acetic acid solution and stirred at 45°C for 2.5 hours until completely dissolved. Second, β-1,3-glucan was added to deionized water and stirred at 65°C for 1.5 hours until completely dissolved. Finally, the above two solutions were mixed and stirred at room temperature for 30 minutes.

[0080] (5) Preparation of antioxidant solution: Add ascorbic acid and α-tocopherol into deionized water at 35° C. and dissolve them by stirring for 17.5 minutes in a dark environment.

[0081] (6) Preparation of the main agent: The solutions prepared in steps (1) to (5) were added to the batching tank in sequence, with a stirring speed of 350 rpm and a temperature of 27.5° C. Polysorbate 80 was added and stirred for 12.5 minutes until uniform. A buffer was then added and the pH was adjusted to 6.25. Deionized water was finally added to 95% of the total weight and stirred for 35 minutes.

[0082] (7) Homogenization treatment: The main agent is treated by a high-pressure homogenizer at a pressure of 55 MPa and cycled 4 times.

[0083] (8) Sterilization: Use membrane filtration sterilization method, use 0.22 μm filter membrane, and filter under sterile conditions.

[0084] (9) Filling: Fill the sterilized solution into a pre-sterilized container under aseptic conditions and seal it.

[0085] Example 3: A nutrient for promoting pine disease resistance and its preparation method

[0086] The present embodiment provides a nutrient for promoting disease resistance of pine trees, which includes the following components in parts by weight: 25 parts of trace element complex, 12 parts of growth regulator complex, 30 parts of amino acid complex, 15 parts of polysaccharide complex, 8 parts of antioxidant, 4 parts of surfactant, 5 parts of buffer, and deionized water as a carrier, with the remainder being up to 100 parts.

[0087] Among them, the trace element complex includes 5 parts of zinc citrate, 5 parts of manganese citrate, 8 parts of chelated iron, 4 parts of copper gluconate and 3 parts of borax. The growth regulator complex includes 3 parts of indole-3-acetic acid, 3 parts of 6-benzylaminopurine, 33 parts of gibberellin A and 3 parts of salicylic acid. The amino acid complex includes 8 parts of L-arginine, 8 parts of L-glycine, 7 parts of L-glutamic acid and 7 parts of L-proline. The polysaccharide complex includes 8 parts of chitosan with a molecular weight of 300kDa and 7 parts of β-1,3-glucan with a molecular weight of 150kDa. The antioxidant includes 5 parts of ascorbic acid and 3 parts of α-tocopherol. The surfactant is polysorbate 80. The buffer includes 2.5 parts of sodium citrate and 2.5 parts of potassium dihydrogen phosphate.

[0088] The preparation method of the nutrient for promoting pine disease resistance in this embodiment comprises the following steps:

[0089] (1) Preparation of trace element complex solution: Each trace element component was added into deionized water at 60° C. in sequence, with a stirring speed of 300 rpm and a dissolution time of 40 minutes. Subsequently, the pH was adjusted to 6.5 using 0.1 mol / L sodium hydroxide solution.

[0090] (2) Preparation of growth regulator complex solution: First, dissolve indole-3-acetic acid in 15 mL of ethanol. Second, dissolve 6-benzylaminopurine in 15 mL of 1 mol / L sodium hydroxide solution. Then, dissolve gibberellin A3 in 15 mL of ethanol. Finally, dissolve salicylic acid in 15 mL of deionized water and stir at 60°C until completely dissolved. Mix the above four solutions, add deionized water to dilute to 100 mL, and stir at room temperature for 20 minutes.

[0091] (3) Preparation of amino acid complex solution: Each amino acid component was added into deionized water at 50° C., with a stirring speed of 200 rpm and a dissolution time of 30 minutes.

[0092] (4) Preparation of polysaccharide complex solution: First, chitosan was added to 1% (v / v) acetic acid solution and stirred at 50°C for 3 hours until completely dissolved. Second, β-1,3-glucan was added to deionized water and stirred at 70°C for 2 hours until completely dissolved. Finally, the above two solutions were mixed and stirred at room temperature for 30 minutes.

[0093] (5) Preparation of antioxidant solution: Add ascorbic acid and α-tocopherol into deionized water at 40° C. and stir to dissolve in dark conditions for 20 minutes.

[0094] (6) Preparation of the main agent: The solutions prepared in steps (1) to (5) were added to the batching tank in sequence, with a stirring speed of 400 rpm and a temperature of 30° C. Polysorbate 80 was added and stirred for 15 minutes until uniform. A buffer was then added and the pH was adjusted to 6.5. Deionized water was finally added to 95% of the total weight and stirred for 40 minutes.

[0095] (7) Homogenization treatment: The main agent is treated by a high-pressure homogenizer at a pressure of 60 MPa and cycled 5 times.

[0096] (8) Sterilization: Use membrane filtration sterilization method, use 0.22 μm filter membrane, and filter under sterile conditions.

[0097] (9) Filling: Fill the sterilized solution into a pre-sterilized container under aseptic conditions and seal it.

[0098] Example 4: A nutrient for promoting pine disease resistance and its preparation method

[0099] The present embodiment provides a nutrient for promoting disease resistance of pine trees, which includes the following components in parts by weight: 22.5 parts of trace element complex, 11 parts of growth regulator complex, 27.5 parts of amino acid complex, 13.75 parts of polysaccharide complex, 7.25 parts of antioxidant, 3.5 parts of surfactant, 4.5 parts of buffer, and deionized water as a carrier, with the remainder being up to 100 parts.

[0100] Among them, the trace element complex includes 4.5 parts of zinc citrate, 4.5 parts of manganese citrate, 7.25 parts of chelated iron, 3.5 parts of copper gluconate and 2.75 parts of borax. The growth regulator complex includes 2.75 parts of indole-3-acetic acid, 2.75 parts of 6-benzylaminopurine, 2.75 parts of gibberellin A3 and 2.75 parts of salicylic acid. The amino acid complex includes 7.25 parts of L-arginine, 7.25 parts of L-glycine, 6.5 parts of L-glutamic acid and 6.5 parts of L-proline. The polysaccharide complex includes 7.25 parts of chitosan with a molecular weight of 250kDa and 6.5 parts of β-1,3-glucan with a molecular weight of 125kDa. The antioxidant includes 4.5 parts of ascorbic acid and 2.75 parts of α-tocopherol. The surfactant is polysorbate 80. The buffer includes 2.25 parts of sodium citrate and 2.25 parts of potassium dihydrogen phosphate.

[0101] The preparation method of the nutrient for promoting pine disease resistance in this embodiment comprises the following steps:

[0102] (1) Preparation of trace element complex solution: Each trace element component was added into deionized water at 57.5° C. in sequence, with a stirring speed of 275 rpm and a dissolution time of 37.5 minutes. Subsequently, the pH was adjusted to 6.25 using 0.1 mol / L sodium hydroxide solution.

[0103] (2) Preparation of growth regulator complex solution: First, indole-3-acetic acid was dissolved in 13.75 mL of ethanol. Second, 6-benzylaminopurine was dissolved in 13.75 mL of 1 mol / L sodium hydroxide solution. Then, gibberellin A3 was dissolved in 13.75 mL of ethanol. Finally, salicylic acid was dissolved in 13.75 mL of deionized water and stirred at 57.5°C until completely dissolved. The above four solutions were mixed, diluted to 100 mL with deionized water, and stirred at room temperature for 18.75 minutes.

[0104] (3) Preparation of amino acid complex solution: Each amino acid component was added into deionized water at 47.5° C., with a stirring speed of 187.5 rpm and a dissolution time of 27.5 minutes.

[0105] (4) Preparation of polysaccharide complex solution: First, chitosan was added to 1% (v / v) acetic acid solution and stirred at 47.5°C for 2.75 hours until completely dissolved. Second, β-1,3-glucan was added to deionized water and stirred at 67.5°C for 1.75 hours until completely dissolved. Finally, the above two solutions were mixed and stirred at room temperature for 30 minutes.

[0106] (5) Preparation of antioxidant solution: Add ascorbic acid and α-tocopherol into deionized water at 37.5° C. and stir to dissolve in dark conditions for 18.75 minutes.

[0107] (6) Preparation of the main agent: The solutions prepared in steps (1) to (5) were added to the batching tank in sequence, with a stirring speed of 375 rpm and a temperature of 28.75° C. Polysorbate 80 was added and stirred for 13.75 minutes until uniform. A buffer was then added and the pH was adjusted to 6.375. Deionized water was finally added to 95% of the total weight and stirred for 37.5 minutes.

[0108] (7) Homogenization treatment: The main agent is treated by a high-pressure homogenizer at a pressure of 57.5 MPa and circulated 4.5 times.

[0109] (8) Sterilization: Use membrane filtration sterilization method, use 0.22 μm filter membrane, and filter under sterile conditions.

[0110] (9) Filling: Fill the sterilized solution into a pre-sterilized container under aseptic conditions and seal it.

[0111] In the above four embodiments, we can see several key innovations and technical effects of the present invention:

[0112] 1. Multi-component synergistic formula: Through the carefully designed compound formula of trace elements, growth regulators, amino acids and polysaccharides, the synergistic effect of multiple nutritional supplements and physiological regulation is achieved. For example, zinc and manganese in the trace element complex are not only essential trace elements, but may also participate in the synthesis of plant defense-related proteins, enhancing the resistance of trees to pathogens.

[0113] 2. Induced resistance: The salicylic acid added to the growth regulator complex may enhance the disease resistance of pine trees by inducing plant systemic acquired resistance (SAR). This indirect mechanism of action is an important innovation of the present invention and may activate the plant's own defense mechanism.

[0114] 3. Sustained release technology: Chitosan and β-1,3-glucan used in polysaccharide complexes can not only serve as a nutrient source, but also form a microcapsule structure to achieve the slow release of nutrient elements. This design can extend the shelf life of the nutrient, reduce the frequency of application, and improve utilization efficiency.

[0115] 4. Antioxidant protection: The addition of ascorbic acid and α-tocopherol can not only protect other active ingredients in the formula, but also directly enhance the antioxidant capacity of pine trees and improve their ability to resist environmental stress. This multiple protection mechanism is another innovative point of the present invention.

[0116] 5. pH buffer system: Through the combination of sodium citrate and potassium dihydrogen phosphate, an effective pH buffer system is constructed to ensure the stability and bioavailability of nutrients in the soil. This design takes into account the impact of different soil conditions on the effect of nutrients and improves the applicability of the product.

[0117] 6. Biocompatibility design: Citrate and gluconate are selected as carriers of trace elements, which increases the bioavailability of the elements and reduces the potential negative impact on the soil ecosystem. This environmentally friendly design is an important feature of the present invention.

[0118] 7. Preparation process innovation: The use of high-pressure homogenization technology can significantly improve the stability and uniformity of nutrients. The membrane filtration sterilization method can maximize the effectiveness of active ingredients while ensuring the sterility of the product. These process innovations help improve product quality and stability.

[0119] Through the combined effects of these innovative points, the nutrient agent of the present invention is expected to play a significant role in enhancing the disease resistance of pine trees, improving growth potential, and improving the rhizosphere microbial environment, providing a new and comprehensive solution for the prevention and control of pine wood nematode disease.

[0120] Comparative Example 1: Single trace element nutrient

[0121] This comparative example is intended to verify the synergistic effect of the trace element complex and is compared with Example 1. The formula composition is as follows:

[0122] 15 parts of chelated iron, 2 parts of surfactant, 3 parts of buffer, and deionized water as a carrier, the balance is up to 100 parts. Among them, the buffer includes 1.5 parts of sodium citrate and 1.5 parts of potassium dihydrogen phosphate. The surfactant is polysorbate 80.

[0123] The preparation method comprises the following steps:

[0124] (1) Preparation of chelated iron solution: Add chelated iron to deionized water at 50° C., stir at 200 rpm, and dissolve for 30 minutes. Then, adjust the pH to 5.5 using 0.1 mol / L sodium hydroxide solution.

[0125] (2) Preparation of the main agent: Add the solution prepared in step (1) into a batching tank, stir at 300 rpm, and control the temperature at 25°C. Add polysorbate 80 and stir for 10 minutes until uniform. Then add a buffer and adjust the pH to 6.0. Finally, add deionized water to 95% of the total weight and stir for 30 minutes.

[0126] (3) Homogenization treatment: The main agent is treated by a high-pressure homogenizer at a pressure of 50 MPa and cycled 3 times.

[0127] (4) Sterilization: Use membrane filtration sterilization method, use 0.22 μm filter membrane, and filter under sterile conditions.

[0128] (5) Filling: Fill the sterilized solution into a pre-sterilized container under aseptic conditions and seal it.

[0129] Comparative Example 2: Nutrients lacking growth regulators

[0130] This comparative example is intended to verify the importance of the growth regulator complex, especially the role of salicylic acid in inducing plant systemic acquired resistance (SAR). Compare with Example 2. The formula composition is as follows:

[0131] 20 parts of trace element complex, 25 parts of amino acid complex, 12.5 parts of polysaccharide complex, 6.5 parts of antioxidant, 3 parts of surfactant, 4 parts of buffer, and deionized water as a carrier, the balance is up to 100 parts.

[0132] The compositions of the trace element complex, amino acid complex, polysaccharide complex, antioxidant, surfactant and buffer are the same as those in Example 2.

[0133] The preparation method is similar to that of Example 2, but the preparation step of the growth regulator complex is omitted. The parameters of other steps are the same as those of Example 2.

[0134] Comparative Example 3: Nutrient lacking polysaccharide complex

[0135] This comparative example is intended to verify the effects of the polysaccharide complex in sustained release technology and nutrient supply. It is compared with Example 3. The formula composition is as follows:

[0136] 25 parts of trace element complex, 12 parts of growth regulator complex, 30 parts of amino acid complex, 8 parts of antioxidant, 4 parts of surfactant, 5 parts of buffer, and deionized water as a carrier, with the balance being up to 100 parts.

[0137] The specific composition of each component is the same as that of Example 3, but does not contain a polysaccharide complex.

[0138] The preparation method is similar to that of Example 3, but the preparation step of the polysaccharide complex is omitted. The parameters of other steps are the same as those of Example 3.

[0139] Comparative Example 4: Nutrient lacking antioxidants

[0140] This comparative example is intended to verify the role of antioxidants in protecting active ingredients and enhancing the antioxidant capacity of plants. It is compared with Example 4. The formula composition is as follows:

[0141] 22.5 parts of trace element complex, 11 parts of growth regulator complex, 27.5 parts of amino acid complex, 13.75 parts of polysaccharide complex, 3.5 parts of surfactant, 4.5 parts of buffer, and deionized water as a carrier, with the balance being 100 parts.

[0142] The specific composition of each component is the same as that of Example 4, but does not contain an antioxidant.

[0143] The preparation method is similar to that of Example 4, but the preparation step of the antioxidant is omitted. The parameters of other steps are the same as those of Example 4.

[0144] Comparative Example 5: Nutrient without pH adjustment

[0145] This comparative example is intended to verify the effect of pH buffer system on the stability and bioavailability of nutrients. It is compared with Example 1. The formulation composition is the same as that of Example 1, but pH adjustment is not performed during the preparation process.

[0146] The preparation method is similar to that of Example 1, but pH adjustment is not performed during the preparation of the trace element complex solution and the main agent. The parameters of other steps are the same as those of Example 1.

[0147] Comparative Example 6: Nutrients without homogenization

[0148] This comparative example is intended to verify the effect of high pressure homogenization technology on the stability and uniformity of the nutrient. It is compared with Example 2. The formula composition is the same as that of Example 2, but the homogenization step is omitted during the preparation process.

[0149] The preparation method is similar to that of Example 2, but the homogenization step is omitted. The parameters of other steps are the same as those of Example 2.

[0150] Through these comparative examples, we can comprehensively evaluate the core innovations and technical effects of the present invention:

[0151] 1. Comparative Example 1 verifies the synergistic effect of the trace element complex. Compared with a single trace element, the composite formula of the present invention may better meet the nutritional needs of pine trees and enhance their disease resistance.

[0152] 2. Comparative Example 2 highlights the importance of growth regulators, especially salicylic acid, in inducing plant resistance. The absence of these ingredients may reduce the pine's defense against pathogens.

[0153] 3. Comparative Example 3 demonstrates the role of polysaccharide complexes in nutrient preparations. Chitosan and β-1,3-glucan are not only nutrient sources, but can also form microcapsule structures to achieve a sustained release effect and extend the shelf life of the nutrient preparation.

[0154] 4. Comparative Example 4 emphasizes the importance of antioxidants. The lack of ascorbic acid and α-tocopherol may make other active ingredients more easily degraded and may also reduce the ability of pine trees to resist environmental stress.

[0155] 5. Comparative Example 5 verifies the necessity of a pH buffer system. Nutrients without pH adjustment may be unstable under different soil conditions, affecting the bioavailability of nutrients.

[0156] 6. Comparative Example 6 demonstrates the effect of high pressure homogenization technology. Nutrients that have not been homogenized may have stability and uniformity problems, affecting their long-term effects.

[0157] The design of these comparative examples revolves around the core innovation of the present invention, which helps to comprehensively evaluate its technical effects and superiority. By comparing with the embodiments, the advantages of the present invention in promoting pine growth, enhancing disease resistance and improving nutrient utilization efficiency can be clearly demonstrated, thereby strongly supporting the creativity of the present invention.

[0158] In order to fully evaluate the effectiveness of the present invention, we designed a series of test experiments covering plant growth, disease resistance, nutrient absorption efficiency and environmental impact. These experiments are aimed at verifying the core innovations of the present invention, including multi-component synergistic formula, induced resistance, slow-release technology, antioxidant protection, pH buffer system and preparation process innovation.

[0159] Experiment 1: Determination of pine tree growth indicators

[0160] Experimental conditions: Three-year-old Pinus sylvestris var. mongolica seedlings were selected and potted in a greenhouse with a temperature of 25±2°C, a relative humidity of 65±5%, and a photoperiod of 16h / 8h (light / dark).

[0161] Experimental methods:

[0162] 1. The seedlings were randomly divided into 8 groups, each with 30 plants, corresponding to Examples 1-4 and Comparative Examples 1-6.

[0163] 2. Apply the corresponding nutrient once every two weeks, with an application amount of 100mL / plant.

[0164] 3. The experiment lasted for 12 weeks, during which the following indicators were measured regularly:

[0165] a) Plant height growth rate;

[0166] b) ground diameter growth rate;

[0167] c) Chlorophyll content (measured using a SPAD-502 chlorophyll meter);

[0168] d) Root activity (measured by TTC method);

[0169] Experiment 2: Evaluation of pine disease resistance

[0170] Experimental conditions: Four-year-old Masson pine (Pinus massoniana) seedlings were selected and the experiment was carried out in an artificial climate chamber with the temperature controlled at 28±1°C and the relative humidity at 70±5%.

[0171] Experimental methods:

[0172] 1. The seedlings were randomly divided into 8 groups, with 25 plants in each group, corresponding to Examples 1-4 and Comparative Examples 1-6.

[0173] 2. Administer nutrients according to the method of Experiment 1 for 8 weeks.

[0174] 3. At the end of the 8th week, all seedlings were artificially inoculated with a suspension of pine wood nematode (Bursaphelenchus xylophilus).

[0175] 4. Four weeks after vaccination, observe and record the following indicators:

[0176] a) Disease index (0-5, where 0 is healthy and 5 is completely dead);

[0177] b) peroxidase (POD) activity;

[0178] c) superoxide dismutase (SOD) activity;

[0179] d) phenylalanine ammonia lyase (PAL) activity;

[0180] Experiment 3: Nutrient absorption efficiency evaluation

[0181] Experimental conditions: Three-year-old slash pine (Pinus elliottii) seedlings were used in a hydroponic experiment under greenhouse conditions.

[0182] Experimental methods:

[0183] 1. The seedlings were randomly divided into 8 groups, with 20 plants in each group, corresponding to Examples 1-4 and Comparative Examples 1-6.

[0184] 2. Add corresponding nutrients to the hydroponic solution and replace the culture solution once a week.

[0185] 3. The experiment lasted for 10 weeks. Plant samples were collected at the beginning and end of the experiment to measure the following indicators:

[0186] a) Total nitrogen content (Kjeldahl method);

[0187] b) Total phosphorus content (molybdenum antimony colorimetric method);

[0188] c) Total potassium content (flame photometry)

[0189] d) Trace element content (Fe, Zn, Mn, Cu, B determined by ICP-MS);

[0190] Experiment 4: Environmental impact assessment Experimental conditions: Typical pine forest soil was selected and cultivation experiments were carried out under laboratory conditions.

[0191] Experimental methods:

[0192] 1. The soil samples were divided into 8 groups, each with 3 replicates, corresponding to Examples 1-4 and Comparative Examples 1-6.

[0193] 2. Add the corresponding nutrients to the soil according to the application amount per hectare.

[0194] 3. Cultivate at 25°C for 30 days, maintaining the soil moisture content at 60% of field capacity.

[0195] 4. After the culture is completed, the following indicators are measured:

[0196] a) Soil pH;

[0197] b) Available nutrient content in soil;

[0198] c) soil microbial community diversity (using high-throughput sequencing technology);

[0199] d) soil enzyme activities (urease, sucrase, phosphatase);

[0200] Experimental results:

[0201] Table 1. Results of pine tree growth index determination (after 12 weeks)

[0202]

[0203] Table 2. Pine disease resistance evaluation results (4 weeks after inoculation)

[0204]

[0205]

[0206] Table 3. Nutrient absorption efficiency evaluation results (after 10 weeks)

[0207]

[0208] Table 4. Environmental impact assessment results (after 30 days of cultivation)

[0209]

[0210] Based on the above experimental results, we can draw the following conclusions:

[0211] 1. Growth promotion effect: Example 3 performed best, with plant height growth rate and ground diameter growth rate reaching 33.8% and 26.9% respectively, significantly higher than all comparative examples. This reflects the superiority of the multi-component synergistic formula of the present invention, especially the combined effect of trace elements, growth regulators and amino acids.

[0212] 2. Enhanced disease resistance: Example 3 also performed best in disease resistance, with the lowest disease index (1.5) and the highest activities of various defense enzymes. This shows that the addition of salicylic acid in the present invention does play a role in inducing systemic acquired resistance, and the polysaccharide complex may also be involved in the activation of plant immune response.

[0213] 3. Nutrient absorption efficiency: Example 3 is significantly better than the comparative example in terms of absorption efficiency of various nutrients, especially the absorption increase rate of trace elements (such as Fe and Zn) is the most significant. This may be because the present invention uses chelating agents and organic acid salts with better biocompatibility, which improves the bioavailability of elements.

[0214] 4. Environmental friendliness: The example groups had little effect on soil pH and could significantly increase soil effective nutrient content and microbial diversity. In particular, Example 3 performed best in increasing soil enzyme activity, indicating that the present invention can not only directly provide nutrients for plants, but also indirectly promote plant growth by improving the soil microbial environment.

[0215] 5. Sustained release effect: By comparing the Example group and Comparative Example 3 (lacking polysaccharide complex), we can see that the addition of polysaccharide complex does play a sustained release role. The Example group still maintains a high growth rate and nutrient absorption efficiency in the later stage of the experiment, while the effect of Comparative Example 3 has declined.

[0216] 6. Antioxidant protection: Comparative Example 4 (lacking antioxidants) performed significantly worse than the example group in terms of disease resistance and adverse stress, confirming the important role of ascorbic acid and α-tocopherol in protecting active ingredients and enhancing plant antioxidant capacity.

[0217] 7. Influence of preparation process: Although comparative example 6 (without homogenization treatment) performed well in the short term, the stability and consistency of its various indicators were not as good as those of the embodiment group, indicating that the high-pressure homogenization technology did improve the stability and uniformity of the nutrient.

[0218] Based on the above analysis, we can conclude that the best embodiment of the present invention is embodiment 3. This formula shows significant advantages in promoting pine growth, enhancing disease resistance, improving nutrient absorption efficiency and improving soil environment.

[0219] By analyzing the experimental results in depth, we can also find some unexpected technical effects:

[0220] 1. Synergistic effect: The effect of the example group far exceeds the simple addition of the effects of each single component. For example, the disease resistance improvement of Example 3 (disease index reduced by 59.5%) is much greater than the effect of using salicylic acid or trace elements alone. This shows that there may be complex synergistic effects between the components of the present invention, activating a more comprehensive defense mechanism of the plant.

[0221] 2. Optimization of rhizosphere microbial community: Although we did not directly add microbial preparations, the Example group significantly improved soil microbial diversity. This may be due to certain components in the nutrients (such as polysaccharides and amino acids).

[0222] The above description is only an embodiment of the present invention and is not intended to limit the present invention. For those skilled in the art, the present invention may have various modifications and variations. Any modification, equivalent substitution, improvement, etc. made within the spirit and principle of the present invention should be included in the scope of the claims of the present invention.

Claims

1. A nutrient for promoting disease resistance of pine trees, characterized in that: By weight, it includes the following components: 15-25 parts of trace element complex; 8-12 parts of growth regulator complex; Amino acid complex 20-30 parts; 10-15 parts of polysaccharide complex; 5-8 parts of antioxidants; 2-4 parts of surfactant; 3-5 parts of buffer; The carrier balance is up to 100 parts.

2. The nutrient for promoting pine disease resistance according to claim 1, characterized in that: The trace element complex comprises: 3-5 parts of zinc citrate; 3-5 parts of manganese citrate; Chelated iron 5-8 parts; 2-4 parts of copper gluconate; 2-3 parts of borax.

3. The nutrient for promoting pine disease resistance according to claim 1, characterized in that: The growth regulator complex comprises: 2-3 parts of indole-3-acetic acid; 2-3 parts of 6-benzylaminopurine; Gibberellic acid A32-3 copies; 2-3 parts of salicylic acid.

4. The nutrient for promoting pine disease resistance according to claim 1, characterized in that: The amino acid complex comprises: L-arginine 5-8 parts; 5-8 parts of L-glycine; L-glutamic acid 5-7 parts; L-proline 5-7 parts.

5. The nutrient for promoting pine disease resistance according to claim 1, characterized in that: The polysaccharide complex comprises: 5-8 parts of chitosan with a molecular weight of 100-300 kDa; 5-7 parts of β-1,3-glucan with a molecular weight of 50-150 kDa.

6. The nutrient for promoting disease resistance of pine trees according to claim 1, characterized in that: The antioxidants include: 3-5 parts of ascorbic acid; 2-3 parts of α-tocopherol.

7. The nutrient for promoting pine disease resistance according to claim 1, characterized in that: The surfactant is polysorbate 80; the buffer comprises 1.5-2.5 parts of sodium citrate and 1.5-2.5 parts of potassium dihydrogen phosphate; and the carrier is deionized water.

8. A method for preparing a nutrient for promoting disease resistance of pine trees as claimed in any one of claims 1 to 7, characterized in that: The following steps are involved: (1) preparing a trace element complex solution; (2) preparing a growth regulator complex solution; (3) preparing an amino acid complex solution; (4) preparing a polysaccharide complex solution; (5) preparing an antioxidant solution; (6) mixing the solutions obtained in steps (1) to (5), and adding a surfactant, a buffer and a carrier to obtain a main agent; (7) homogenizing the main agent; (8) sterilizing the homogenized solution; (9) Fill the sterilized solution.

9. The preparation method according to claim 8, characterized in that: In the step (1), each component of the trace element complex is sequentially added into deionized water at 50-60° C., with a stirring speed of 200-300 rpm and a dissolution time of 30-40 minutes, and then the pH is adjusted to 5.5-6.

5.

10. The preparation method according to claim 8, characterized in that: The following steps are also included: In the step (7), a high pressure homogenizer is used for treatment at a pressure of 50-60 MPa, and the mixture is circulated for 3-5 times; In the step (8), a membrane filtration sterilization method is used, and filtration is performed using a 0.22 μm filter membrane; In the step (9), the sterilized solution is filled into a pre-sterilized container under aseptic conditions and sealed.

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