Special energy-type immunizing agent for plant blight and verticillium wilt and preparation method of special energy-type immunizing agent

Abstract

The invention discloses a special energy type immunizing agent for plant blight and verticillium wilt, and the special energy type immunizing agent for plant blight and verticillium wilt is prepared from the following components: 200 to 1000 parts of radix et rhizoma rhei water extract, 100 to 1500 parts of plant immune yield-increasing protein Pulvtong, 200 to 600 parts of gibberellin, 130 to 450 parts of sodium p-hydroxybenzoate and 120 to 340 parts of paenibacillus polymyxa. The invention also discloses a preparation method of the energy-type immunizing agent special for plant blight and verticillium wilt. The energy-type immunizing agent is prepared by mixing the rhubarb water extract, the plant immune yield-increasing protein Pulvtong, gibberellin and paenibacillus polymyxa, so that the purposes of improving the autoimmunity of plants, improving the antibacterial and antiviral capabilities, promoting the growth of the plants, increasing the yield and enhancing the stress resistance of the plants are achieved, all-around protection is provided for the plants, and the economic benefits of the plants are increased. The composition can effectively prevent and treat wilt and verticillium wilt, can improve the overall health level of plants, does not harm human health and has no pesticide residue problem, and the quality of agricultural products is better after the composition is used.

Description

Technical Field

[0001] The invention belongs to the technical field of plant medicines, and in particular relates to an energy-type immune agent specially used for plant verticillium wilt and a preparation method thereof. Background Art

[0002] Verticillium wilt is a plant vascular disease mainly caused by Verticillium dahliae and Verticillium nigra, which causes serious damage to cotton, Solanaceae and many other plants. In addition to hyphae, Verticillium wilt also forms conidia and sclerotia. Sclerotia are persistent organs generated by the spore division of hyphae. They are produced in large quantities on diseased plants and overwinter in the soil in the form of sclerotia. These sclerotia are spread together with the residue of diseased plants by wind and rain and spread to the surrounding areas. Infected seedlings can also bring the pathogen to non-infected areas. When the seedlings are transplanted, the sclerotia will germinate. Mycelium grows from the tubes and infects the plant from the top of the root and the wound. The pathogens that invade the plant body mainly reproduce in the ducts, hindering the rise of water and producing toxins, causing the plant to wilt. At present, chemical control with pesticides is the main means of preventing and controlling wilt disease, but excessive use of pesticides will lead to excessive pesticide residues in agricultural products, affecting human health. Long-term use of chemical pesticides will pollute the soil, water sources and other environments, and destroy the ecological balance. In this regard, we have proposed an energy-type immune agent specifically for plant wilt disease and a preparation method thereof. Summary of the Invention

[0003] In view of the deficiencies in the prior art, the present invention provides an energy-type immune agent specifically for plant Verticillium wilt and a preparation method thereof to solve the above technical problems.

[0004] To achieve the above-mentioned purpose, the present invention provides the following technical solution: an energy-type immune agent specifically for plant fusarium wilt, comprising the following ingredients: 200-1000 parts of rhubarb water extract, 100-1500 parts of plant immune production-increasing protein Pu Lu Tong, 200-600 parts of gibberellins, 130-450 parts of sodium p-hydroxybenzoate and 120-340 parts of Bacillus polymyxa.

[0005] Preferably, the invention is specifically composed of the following ingredients: 640 parts of rhubarb water extract, 1120 parts of plant immune production-increasing protein Pulutong, 320 parts of gibberellins, 256 parts of sodium p-hydroxybenzoate and 176 parts of Paenibacillus polymyxa.

[0006] Preferably, the rhubarb aqueous extract is extracted from rhubarb by a water extraction method, comprising placing the coarsest powder of rhubarb medicinal material into a container and soaking it in deionized water, heating the soaked rhubarb medicinal material at 95° C. for 1 hour in a water bath and subjecting it to reflux extraction, filtering the solution with gauze after it cools to remove solid impurities, repeating the heating and extraction steps five times, and combining the filtrates to increase the concentration to obtain the rhubarb aqueous extract.

[0007] Preferably, the plant immune yield-increasing protein, Pu Lu Tong, is a protein biological pesticide, prepared by culturing Alternaria alternata and Colletotrichum thunbergii on a blood agar plate medium for 60 hours, inoculating the bacteria into a solid fermentation medium for primary fermentation, stirring at a speed of 60 rpm during the culture process to obtain a fermentation liquid, and then fermenting the fermentation liquid in the solid fermentation medium for three times, adding a mixed solution of dipotassium hydrogen phosphate and potassium dihydrogen phosphate, a protein protective agent and humic acid to the fermentation liquid, stirring and mixing, and then spray-drying to obtain the plant immune yield-increasing protein, Pu Lu Tong.

[0008] Preferably, gibberellin is a plant hormone. The gibberellin strain is inoculated onto a slant culture medium and activated at 25°C-28°C. Mycelial blocks are picked from the activated slant strain and inoculated into a shake flask filled with liquid culture medium. Oscillation culture is carried out at the same temperature to allow the strain to multiply in large quantities. The seed liquid in the shake flask is introduced into a fermenter. The temperature in the fermenter is controlled at 25°C-28°C, the pH value is within the range of 4.5-5.5, the fermentation is stirred at 60 rpm, and the fermentation time is 200 hours. After the fermentation is completed, the bacteria and impurities in the fermentation liquid are removed by centrifugation to obtain gibberellin.

[0009] Preferably, sodium p-hydroxybenzoate is a compound, which is prepared by mixing phenol and a 40% potassium hydroxide solution in a reaction pot, stirring at 70 rpm for 0.5 h at 100° C. until the free base concentration of the potassium phenolate solution is 0.3%-1.2%, heating the reaction solution for atmospheric dehydration, switching to reduced pressure dehydration when the internal temperature reaches 140° C., distilling water at a pressure of 10.6 kPa for 0.5 h-1 h until the internal temperature reaches above 170° C., adding phenol as a solvent for azeotropic dehydration, and completing the dehydration when the temperature reaches 200° C. and the pressure reaches 2.67 kPa, thereby obtaining a composite salt of potassium phenolate and phenol. The composite salt is further heated to 220°C-230°C, and purified anhydrous carbon dioxide is introduced, the pressure is maintained at 0.5 MPa, and the reaction is carried out for 2.5 hours. The temperature is then lowered to 200°C, phenol is added, and the mixture is stirred at 60 rpm for 30 minutes while maintaining the temperature. The phenol is then recovered under reduced pressure for a second carboxylation, and carbon dioxide is introduced for 2 hours. After the carboxylation is completed, the phenol is recovered and cooled to below 180°C, and water is added to dissolve the recovered phenol to obtain a carboxylated solution. Sulfuric acid is gradually added to the carboxylated solution to neutralize the solution to a pH of 6.7 at a temperature below 70°C. The solution is cooled and filtered to remove potassium sulfate to obtain a crude filter cake, which is then recrystallized with water and decolorized with activated carbon to obtain sodium p-hydroxybenzoate.

[0010] Preferably, Paenibacillus polymyxa is a bacterium that is prepared by inoculating Paenibacillus polymyxa strains into a sterile nutrient agar medium to form a suspension of the strains, and performing slant inoculation on a solid culture medium, i.e., evenly spreading the suspension of the strains on the slant surface of the culture medium, maintaining the growth environment temperature at 30°C-32°C, and culturing for 24h-48h to obtain Paenibacillus polymyxa.

[0011] A method for preparing a special energy-type immune agent for plant Verticillium wilt, the preparation steps comprising:

[0012] S1. Prepare the corresponding number of materials;

[0013] S2, placing the rhubarb water extract, the plant immune production-increasing protein Pulutong, and gibberellin in a low-temperature magnetic stirrer and stirring evenly to form a mixed solution;

[0014] S3. After mixing evenly, add Paenibacillus polymyxa to the mixture and continue stirring evenly;

[0015] S4. Add acid-base regulator to adjust the pH value of the mixture to 5-9;

[0016] S5. Inject the prepared immune agent into a vial, seal it, and store it at low temperature.

[0017] Preferably, in step S2, the temperature of the low-temperature magnetic stirrer is set at 10°C-15°C, the stirring speed is set at 100 rpm, and the stirring time is 1 hour. In step S3, the temperature of the low-temperature magnetic stirrer is set at 4°C-6°C, the stirring speed is set at 60 rpm, and the stirring time is 0.5 hour.

[0018] Preferably, the storage temperature of the immune agent in step S5 is 4°C-6°C.

[0019] Compared with the prior art, the present invention has the following beneficial effects:

[0020] This application prepares an energy-type immune agent by mixing rhubarb water extract, plant immune yield-increasing protein Prolutong, gibberellin and Bacillus polymyxa, so as to achieve the purpose of enhancing the plant's own immunity, improving the antibacterial and antiviral capabilities, promoting plant growth and yield, and enhancing plant stress resistance, providing all-round protection for plants, not only effectively preventing and treating fusarium wilt, but also improving the overall health level of plants, without harming human health, without pesticide residue problems, and with better quality of agricultural products after use. DETAILED DESCRIPTION

[0021] The following will be combined with the embodiments of the present invention to clearly and completely describe the technical solutions in the embodiments of the present invention. Obviously, the embodiments described 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 efforts are within the scope of protection of the present invention.

[0022] The present invention provides a technical solution: an energy-type immune agent specially used for treating plant fusarium wilt, which is composed of the following components: 200-1000 parts of rhubarb water extract, 100-1500 parts of plant immune production-increasing protein Pulutong, 200-600 parts of gibberellins, 130-450 parts of sodium p-hydroxybenzoate and 120-340 parts of Paenibacillus polymyxa.

[0023] Furthermore, the invention is specifically composed of the following ingredients: 640 parts of rhubarb water extract, 1120 parts of plant immune production-increasing protein Pulutong, 320 parts of gibberellins, 256 parts of sodium p-hydroxybenzoate and 176 parts of Paenibacillus polymyxa.

[0024] Furthermore, the rhubarb aqueous extract is extracted from rhubarb by a water extraction method, wherein the rhubarb medicinal material is washed, dried and crushed to form the coarsest powder, and the coarsest powder of the rhubarb medicinal material is placed in a container and soaked in deionized water. The deionized water is used to reduce the influence of impurities in the water on the extract and ensure the purity of the extract. During the soaking process, the soluble components in the rhubarb medicinal material will gradually dissolve in the water to form a preliminary extract. In order to accelerate the extraction process and improve the extraction efficiency, a water bath is used to heat the soaked rhubarb medicinal material, and the soaked rhubarb medicinal material is heated at 95° C. for 1 hour using a water bath for reflux extraction. After the solution is cooled, it is filtered with gauze to remove solid impurities, the heating extraction step is repeated 5 times, and the filtrate is combined to increase the concentration to obtain the rhubarb aqueous extract. During the concentration process, the temperature and time need to be strictly controlled to avoid the heat-sensitive loss of the effective ingredients.

[0025] Rhubarb aqueous extract comes from rhubarb, which is the dried root and rhizome of Rheum palmatum, Rheum tanguticum or medicinal rhubarb of the Polygonaceae family. The rhubarb aqueous extract has a 100% antibacterial rate against Verticillium dahliae, the main cause of Verticillium wilt. It can inhibit the germination of Verticillium dahliae spores to form hyphae, destroy the hyphae surface and cause deformity, inhibit the reproduction of pathogenic fungi in plants, and effectively prevent and control Verticillium wilt. Rhubarb aqueous extract can stimulate the expression of plant resistance genes, reduce the content of malondialdehyde in plants, enhance the enzyme activities of PPO and SOD, improve the plant's own stress resistance and promote plant growth.

[0026] Furthermore, the plant immune yield-increasing protein Pulutong protein biological pesticide is cultured by Alternaria alternata and Colletotrichum on blood agar plate medium for 60 hours. The strains will multiply rapidly and produce metabolites that are beneficial to plant growth. After the plate culture is completed, the vigorous bacteria are inoculated into the solid fermentation medium through aseptic operation for primary fermentation culture. The culture medium will provide the carbon source, nitrogen source, inorganic salts and growth factors required for the growth of the bacteria, and at the same time optimize the fermentation conditions to promote the rapid growth of the bacteria and the accumulation of metabolites. During the culture process, the fermentation liquid is obtained by stirring at a speed of 60 rpm, and the fermentation culture of the fermentation liquid in the solid fermentation medium is repeated 3 times. In the culturing and fermentation step, after multiple fermentation cultures are completed, a mixture of dipotassium hydrogen phosphate and potassium dihydrogen phosphate, a protein protective agent, and humic acid are added to the fermentation liquid. The phosphate mixture provides a suitable pH environment and ionic strength for the protein, enhancing the stability of the protein. The protein protective agent can effectively prevent the denaturation and degradation of the protein during subsequent processing. Humic acid, as a natural organic substance, can improve soil structure, promote plant root development, and enhance plant stress resistance. After stirring and mixing, spray drying is performed. The spray drying technology can quickly convert the liquid into a fine granular product while retaining its biological activity and nutrients, thereby producing the plant immune and yield-increasing protein Pu Lu Tong.

[0027] The plant immune yield-increasing protein Pu Lu Tong combines plant immune-inducing protein and amino oligosaccharide. Through mutual synergistic effects, it fully stimulates the plant's immune-inducing resistance potential, and has a highly effective preventive effect on viral, fungal, and bacterial diseases, especially for viral diseases. Whether used as seed treatment, seedbed spraying, or root dipping before transplanting, the plant immune yield-increasing protein Pu Lu Tong can significantly promote the development of crop seedling roots. The fresh weight of the root system can increase by 20%-30% compared with the control, which manifests as deep roots and lush leaves, and strong seedlings. It has the dual effects of promoting cell division and cell elongation, increasing the chlorophyll content and photosynthetic efficiency of plant leaves, and promoting crop nutritional growth. At the same time, it increases the germination rate of pollen and the elongation of pollen tubes, thereby increasing the fruit set rate and fruit setting rate, which is beneficial to increasing yields. It can also improve the drought and cold resistance of crops, and reduce the damage to normal functions of harmful substances produced in the plant body under adverse conditions. It adds a variety of nutrients necessary for crop growth, providing comprehensive protection for the healthy growth of crops.

[0028] Furthermore, gibberellin is a plant hormone. The gibberellin strain is inoculated onto the slant culture medium to provide a suitable living environment for the strain, allowing the gibberellin strain to undergo initial growth and activation. The activation is carried out at 25°C-28°C, and the mycelium blocks are picked from the activated slant strains. The mycelium blocks are required to be healthy, well-grown and rich in active cells. They will be used as seeds for the fermentation process. The mycelium blocks are inoculated into shake flasks filled with liquid culture medium. The liquid culture medium will meet the growth and metabolic needs of gibberellin. Oscillating culture is carried out at the same temperature. Through continuous oscillation, the oxygen and nutrients in the culture medium are evenly distributed, which promotes the rapid reproduction of the strain and the initial synthesis of gibberellin. As the strain reproduces in large numbers, its metabolic activity becomes more and more vigorous, and the content of gibberellin gradually accumulates. At this time, the seed liquid in the shake flask is connected to the fermentation tank. The temperature in the fermentation tank is controlled at 25°C-28°C and the pH value is between 4.5 and 5. .5 range, stirred at 60rpm, and fermented for 200h. During this period, various parameters in the fermentation tank need to be closely monitored, including but not limited to temperature, pH value, dissolved oxygen content and gibberellin accumulation. After the fermentation is completed, the bacteria and impurities in the fermentation liquid are removed by centrifugation to obtain gibberellin. Gibberellic acid can promote stem elongation, germination, breaking of seed dormancy, flowering, and growth and development of fruits, and has a significant yield-increasing effect on most crops. By treating seeds with an appropriate concentration of gibberellin solution, the dormancy period of seeds can be broken early, the germination rate can be increased, and production and income can be increased. Gibberellic acid can also regulate the antioxidant capacity of plants, increase the activity of superoxide dismutase antioxidant enzymes, and protect cells from oxidative damage. When plants suffer from drought and salt stress, gibberellin can regulate the osmotic regulation ability of plants, improve cell stability and tolerance, and help plants adapt to different environmental conditions.

[0029] Furthermore, sodium p-hydroxybenzoate is a compound. By mixing phenol with a 40% solution of potassium hydroxide in a reaction pot, the subsequent reaction can be ensured to proceed smoothly, because potassium hydroxide converts phenol into potassium phenolate, providing an active intermediate for the subsequent carboxylation reaction. In the reaction pot, the mixture is stirred at 70 rpm for 0.5 h at a temperature of 100 ° C. until the free base concentration of the potassium phenolate solution is 0.3%-1.2%. During this process, the free base concentration of the reaction solution needs to be closely monitored and the free base concentration of the potassium phenolate solution is ensured by titration analysis. The reaction solution is heated for atmospheric dehydration. When the internal temperature reaches 140 ° C., it is changed to reduced pressure dehydration. Water is distilled at a pressure of 10.6 kPa for 0.5 h-1 h until the internal temperature reaches above 170 ° C. Solvent phenol is added for azeotropic dehydration. The dehydration is terminated when the temperature reaches 200 ° C. and the pressure reaches 2.67 kPa to obtain a composite salt of potassium phenolate and phenol. The composite salt is further heated to Purified anhydrous carbon dioxide is introduced at 220°C-230°C, the pressure is maintained at 0.5MPa, the reaction is carried out for 2.5h, the temperature is lowered to 200°C, phenol is added, the mixture is kept warm and stirred for 30 minutes at a stirring speed of 60rpm, and then the phenol is recovered under reduced pressure for a second carboxylation, carbon dioxide is introduced for 2h to ensure the complete formation of potassium p-hydroxybenzoate, and after the carboxylation is completed, the phenol is recovered and cooled to below 180°C, and water is added to dissolve it to obtain a carboxylated liquid, sulfuric acid is gradually added to the carboxylated liquid and neutralized to a pH of 6.7 at a temperature below 70°C to avoid local overheating, the crude filter cake is cooled and filtered to remove potassium sulfate to obtain a crude filter cake, and in order to further improve the purity and quality of the product, the crude filter cake is recrystallized with water and decolorized with activated carbon to obtain sodium p-hydroxybenzoate, which promotes the division, elongation and differentiation of plant cells, enhances the tension and elasticity of cell walls, and thus promotes the growth and development of plants, and can enhance the stability and durability of the immune agent in the immune agent.

[0030] Furthermore, Bacillus polymyxa is a bacterium. The Bacillus polymyxa strain is inoculated into a sterile nutrient agar medium, and the strain is evenly dispersed in the medium through stirring to prepare a strain suspension. Slant inoculation is performed on the solid culture medium. Slant inoculation is a commonly used microbial culture method, which uses the slope formed by the culture medium after solidification to provide a larger growth area for the strain. That is, the suspension of the strain is evenly spread on the slope of the culture medium with a sterile inoculation loop to ensure that the strain can be evenly distributed and attached to the surface of the culture medium. After the inoculation is completed, the culture dish is placed in a constant temperature incubator and the growth environment temperature is maintained at 30°C-32°C. This temperature range is the best condition for the growth of Bacillus polymyxa, which is conducive to the rapid reproduction of the strain and the accumulation of metabolites. The culture time is 24h-48h. Use a sterile inoculation loop to gently scrape the colonies on the slope without destroying the integrity of the culture medium to obtain Bacillus polymyxa. The "using bacteria to treat bacteria" approach has a strong colonization ability in the roots, stems, and leaves of plants, and can prevent pathogens from infecting plants through site competition. Bacillus polymyxa can continuously secrete broad-spectrum antibacterial substances, including but not limited to polymyxins, colistin, and cyclobactin, which can inhibit or kill pathogens, thereby achieving the purpose of preventing and controlling verticillium wilt diseases. During the reproduction process, Bacillus polymyxa can produce plant hormones auxin and mitogen to promote plant growth. By enhancing plant photosynthesis, providing plants with nitrogen sources and degrading P and K complexes into nutrients that can be absorbed and utilized by plants, it further promotes plant growth and increases production. Bacillus polymyxa can increase plant yield and quality, and by enhancing plant disease resistance and stress resistance, it reduces the occurrence of diseases, thereby improving the overall yield and quality of crops. Bacillus polymyxa is a microbial strain that is safe and non-toxic to plants. No matter what concentration is used, it will not cause any phytotoxicity to crops. It is a safe and environmentally friendly biological pesticide.

[0031] A method for preparing a special energy-type immune agent for plant Verticillium wilt, the preparation steps comprising:

[0032] S1. Prepare the corresponding number of materials;

[0033] S2, placing the rhubarb water extract, the plant immune production-increasing protein Pulutong, and gibberellin in a low-temperature magnetic stirrer and stirring evenly to form a mixed solution;

[0034] S3. After mixing evenly, add Paenibacillus polymyxa to the mixture and continue stirring evenly;

[0035] S4. Add acid-base regulator to adjust the pH value of the mixture to 5-9;

[0036] S5. Inject the prepared immune agent into a vial, seal it, and store it at low temperature.

[0037] Furthermore, in step S2, the temperature of the low-temperature magnetic stirrer is set at 10°C-15°C, the stirring speed is set to 100rpm, and the stirring time is 1h. In step S3, the temperature of the low-temperature magnetic stirrer is set at 4°C-6°C, the stirring speed is set to 60rpm, and the stirring time is 0.5h. The low-temperature magnetic stirrer uses the magnetic principle to uniformly stir the liquid sample under low temperature conditions. It consists of two parts: a magnetic stirrer and a magnetic drive. The magnetic stirrer includes a magnetic rotor and a sample solution in a container, and the magnetic drive includes an external motor and a magnetic rod that generates a rotating magnetic field. When the external motor is started, the magnetic rod generates a rotating magnetic field, causing the magnetic rotor to rotate in the container, thereby stirring the solution. It has good temperature control performance and is equipped with a constant temperature system. It can accurately control the sample temperature in a low-temperature environment, which is very important for the preparation of stirring immune agents that need to be carried out under low-temperature conditions.

[0038] Furthermore, the storage temperature of the immunizing agent in step S5 is 4°C-6°C. The low temperature can ensure that the number of living and dead Paenibacillus polymyxa is reduced, inhibit the growth of bacteria and ensure the functionality of the immunizing agent.

[0039] It should be noted that, in this document, relational terms such as first and second, etc., are used only to distinguish one entity or operation from another entity or operation, and do not necessarily require or imply any actual relationship or order between these entities or operations. Moreover, the terms "comprises," "comprising," or any other variations thereof are intended to cover non-exclusive inclusion, such that a process, method, article, or apparatus that includes a list of elements includes not only those elements but also other elements not explicitly listed, or elements inherent to such process, method, article, or apparatus.

[0040] While embodiments of the present invention have been shown and described, it will be appreciated by those skilled in the art that various changes, modifications, substitutions, and variations may be made to these embodiments without departing from the principles and spirit of the invention, and that the scope of the invention is defined by the appended claims and their equivalents.

Claims

1. A special energy-type immune agent for plant Verticillium wilt, characterized in that: The invention is composed of the following ingredients: 200-1000 parts of rhubarb water extract, 100-1500 parts of plant immune production-increasing protein Pulutong, 200-600 parts of gibberellins, 130-450 parts of sodium p-hydroxybenzoate and 120-340 parts of Paenibacillus polymyxa.

2. The energy-type immune agent for plant Verticillium wilt according to claim 1, characterized in that: Specifically, the invention comprises the following ingredients: 640 parts of rhubarb water extract, 1120 parts of plant immune production-increasing protein Prolutong, 320 parts of gibberellins, 256 parts of sodium p-hydroxybenzoate and 176 parts of Paenibacillus polymyxa.

3. The energy-based immune agent for plant Verticillium wilt according to claim 1, characterized in that: The rhubarb aqueous extract is extracted from rhubarb by a water extraction method, wherein the coarsest powder of the rhubarb medicinal material is placed in a container and deionized water is added for soaking, the soaked rhubarb medicinal material is heated at 95°C for 1 hour in a water bath and reflux extracted, the solution is cooled and filtered with gauze to remove solid impurities, the heating extraction step is repeated 5 times, and the filtrate is combined to increase the concentration to obtain the rhubarb aqueous extract.

4. The energy-based immune agent for plant Verticillium wilt according to claim 1, characterized in that: The plant immune and yield-increasing protein, Pu Lu Tong, is a protein biological pesticide. The method comprises the following steps: culturing Alternaria alternata and Colletotrichum spp. under blood agar plate culture medium for 60 hours, inoculating the bacteria into a solid fermentation medium for primary fermentation culture, stirring at a speed of 60 rpm during the culture process to obtain a fermentation liquid, and then fermenting the fermentation liquid in the solid fermentation medium for three times. A mixed solution of dipotassium hydrogen phosphate and potassium dihydrogen phosphate, a protein protective agent and humic acid are added to the fermentation liquid, stirring and mixing, and then spray-drying to obtain the plant immune and yield-increasing protein, Pu Lu Tong.

5. The energy-based immune agent for plant Verticillium wilt according to claim 1, characterized in that: Gibberellic acid is a plant hormone. Gibberellic acid strains are inoculated onto a slant culture medium and activated at 25°C-28°C. Mycelial blocks are picked from the activated slant culture medium and inoculated into a shake flask filled with liquid culture medium. Oscillation culture is carried out at the same temperature to allow the strain to multiply in large quantities. The seed liquid in the shake flask is introduced into a fermentation tank. The temperature in the fermentation tank is controlled at 25°C-28°C, the pH value is within the range of 4.5-5.5, and the fermentation is stirred at 60 rpm for 200 hours. After the fermentation is completed, the bacteria and impurities in the fermentation liquid are removed by centrifugation to produce gibberellin.

6. The energy-based immune agent for plant Verticillium wilt according to claim 1, characterized in that: Sodium p-hydroxybenzoate is a compound, which is prepared by mixing phenol and 40% potassium hydroxide solution in a reaction pot, stirring at 70 rpm for 0.5 h at 100°C, until the free base concentration of potassium phenolate solution is 0.3%-1.2%, heating the reaction liquid for normal pressure dehydration, and switching to reduced pressure dehydration when the internal temperature reaches 140°C, distilling water at 10.6 kPa pressure for 0.5 h-1 h, until the internal temperature reaches above 170°C, adding solvent phenol for azeotropic dehydration, and completing dehydration when the temperature reaches 200°C and the pressure reaches 2.67 kPa, thereby obtaining a composite salt of potassium phenolate and phenol. The salt mixture is further heated to 220°C-230°C, and purified anhydrous carbon dioxide is introduced, the pressure is maintained at 0.5 MPa, the reaction is carried out for 2.5 hours, the temperature is lowered to 200°C, phenol is added, and the mixture is stirred at 60 rpm for 30 minutes at a temperature of 100 rpm. The phenol is then recovered under reduced pressure for a second carboxylation, and carbon dioxide is introduced for 2 hours. After the carboxylation is completed, the phenol is recovered and cooled to below 180°C, and water is added to dissolve the recovered phenol to obtain a carboxylated solution. Sulfuric acid is gradually added to the carboxylated solution to neutralize the solution to a pH of 6.7 at a temperature below 70°C. The solution is cooled and filtered to remove potassium sulfate to obtain a crude filter cake, which is then recrystallized with water and decolorized with activated carbon to obtain sodium p-hydroxybenzoate.

7. The energy-based immune agent for plant Verticillium wilt according to claim 1, characterized in that: Paenibacillus polymyxa is a bacterium that can be obtained by inoculating the Paenibacillus polymyxa strain into a sterile nutrient agar medium to prepare a strain suspension, and performing slant inoculation on a solid culture medium, i.e., evenly spreading the strain suspension on the slant surface of the culture medium, maintaining the growth environment temperature at 30°C-32°C, and culturing for 24h-48h.

8. A method for preparing an energy-type immune agent specifically for plant Verticillium wilt, characterized in that: The preparation steps include: S1. Prepare the corresponding number of materials; S2, placing the rhubarb water extract, the plant immune production-increasing protein Pulutong, and gibberellin in a low-temperature magnetic stirrer and stirring evenly to form a mixed solution; S3. After mixing evenly, add Paenibacillus polymyxa to the mixture and continue stirring evenly; S4. Add acid-base regulator to adjust the pH value of the mixture to 5-9; S5. Inject the prepared immune agent into a vial, seal it, and store it at low temperature.

9. The method for preparing the energy-type immune agent for plant Verticillium wilt according to claim 8, characterized in that: In step S2, the temperature of the low-temperature magnetic stirrer is set at 10°C-15°C, the stirring speed is set at 100 rpm, and the stirring time is 1 hour. In step S3, the temperature of the low-temperature magnetic stirrer is set at 4°C-6°C, the stirring speed is set at 60 rpm, and the stirring time is 0.5 hour.

10. The method for preparing the energy-type immune agent for plant Verticillium wilt according to claim 8, characterized in that: The storage temperature of the immune agent in step S5 is 4°C-6°C.