Method for producing organic water-soluble fertilizer by using tagetes fermentation press liquor

Abstract

The application discloses a method for producing organic water-soluble fertilizer by using tagetes fermenting pressure filtrate, and belongs to the field of fertilizers.The method comprises the following steps: preparing tagetes water-soluble fertilizer by adjusting the pH of tagetes fermenting pressure filtrate, separating solid and liquid to obtain liquid, concentrating, cooling and settling to obtain liquid; and preparing rosemary fermenting liquid by fermenting rosemary waste residues by using Trichoderma asperellum, and separating solid and liquid to obtain liquid.The two agricultural wastes are converted into high-value functional fertilizer, and resource circulation and medicine-fertilizer integration are realized.Through biological fermentation, the tagetes water-soluble fertilizer provides growth-promoting nutrition and improves the synthesis of capsaicin, and the rosemary fermenting liquid introduces Trichoderma asperellum live bacteria to specially prevent and treat pepper blight.The application is especially suitable for the planting of peppers, and is also a general type of water-soluble fertilizer.

Description

Technical Field

[0001] This invention belongs to the field of fertilizers, and in particular relates to a method for producing organic water-soluble fertilizer using marigold fermentation filtrate. Background Technology

[0002] In the process of producing lutein from marigolds through fermentation in lutein factories, marigold fermentation filtrate is generated; dry hot air with an outlet temperature of 300-400 degrees Celsius is generated by burning biomass fuel or coal. This dry hot air is then purified, cooled to 80-90 degrees Celsius, and can only be used for drying marigold petal fragments; a large amount of heat is lost throughout the process.

[0003] Because the fermentation filtrate of marigolds has high COD (chemical oxygen demand) and BOD (biochemical oxygen demand), and a low pH level of 3-6, and contains a large amount of organic matter, nitrogen, potassium and other components, if it is discharged directly without treatment, it will pollute the environment and waste useful water-soluble organic matter, amino acids, biochemical fulvic acid, nitrogen, potassium and other fertilizer resources.

[0004] The commonly used methods for treating marigold fermentation filtrate are as follows: 1) Farmland irrigation method – The marigold fermentation filtrate is diluted and used directly to irrigate farmland. This method is subject to limitations regarding the location and timing of fertilization. 2) Treatment method of fermentation wastewater by environmental protection station: Most lutein factories build a small wastewater treatment plant to achieve standard discharge. The cost of treating 1 ton of wastewater is 60-80 yuan.

[0005] 3) Concentrated spray drying method – Concentrate and dry the marigold fermentation filtrate.

[0006] These methods are limited by various conditions and cannot fully utilize marigold fermentation filtrate (e.g.); secondly, they cannot fully utilize marigold fermentation filtrate, but the equipment investment is large and the operating cost is high (e.g., the concentrated spray drying method requires an investment of more than 10 million yuan for a single set of equipment, and the treatment cost of 1 ton of marigold fermentation filtrate exceeds 100 yuan). Their investment and operating costs are difficult for ordinary enterprises to bear, which is not conducive to their promotion and application. Summary of the Invention

[0007] The first objective of this invention is to provide an organic water-soluble fertilizer comprising marigold water-soluble fertilizer and rosemary fermentation liquid, wherein the mass ratio is 2-3:1; The marigold water-soluble fertilizer is obtained by adjusting the pH of marigold fermentation filtrate, separating solid and liquid to obtain liquid, concentrating, cooling and settling to obtain liquid. The marigold fermentation filtrate is the liquid obtained by pressing and filtering after solid-state fermentation pretreatment when producing lutein using the marigold fermentation method. The rosemary fermentation liquid is obtained by fermenting rosemary waste residue with bacteria of accession number CICC13028, followed by solid-liquid separation to obtain liquid. The rosemary residue is the solid waste produced after extracting essential oils.

[0008] Preferably, the preparation of the marigold water-soluble fertilizer includes the following steps: S1: Adjust the pH of the marigold fermentation filtrate to 6.0-6.5; S2: Heat the marigold fermentation filtrate after pH adjustment to boiling and remove foam to obtain boiling liquid; S3: Separate the boiling liquid from the solid and liquid phases and collect the liquid phase; S4: The liquid phase is concentrated by evaporation to a solid content of 50-60%; a concentrated solution is obtained. S5: The concentrate is cooled and settled to separate the solid and liquid phases. The liquid phase is then taken as marigold water-soluble fertilizer.

[0009] Furthermore, it also includes the following steps: S6: Wash the solid phase separated in step S5, wherein the washing solution is returned to step S4; the washed solid phase is used to prepare lutein.

[0010] In a further step S4, the evaporation container containing the liquid phase is cleaned with an acid pickling agent, and the cleaning solution is returned to step S1; the acid pickling agent is hydrochloric acid and / or nitric acid.

[0011] Preferably, the preparation of the rosemary fermented liquid fertilizer includes the following steps: S1: After extracting essential oil, the rosemary is dried at a temperature below 50°C until the moisture content is ≤15%; then it is crushed into particles with a particle size ≤2 mm to obtain rosemary waste residue; S2: Prepare the culture medium according to the following proportions by weight. Rosemary residue, 100 portions; Carbon source, at least 10 parts; Nitrogen source, at least 4 parts; Potassium dihydrogen phosphate, at least 0.2 parts; Magnesium sulfate heptahydrate, at least 0.1 parts; Adjust the water content of the culture medium to between 55-65%, and the initial pH to 5.0-5.5; S3: Activate the bacteria with accession number CICC13028, and then prepare a concentration ≥1×10⁻⁶. 7 CFU / mL spore suspension; S4: Inoculate the spore suspension into the culture medium for aerobic fermentation; the criterion for completion of fermentation is a pH change of ≤0.1 within 12 hours; S5: After fermentation, water is added to the solid material for extraction, and then the filtrate is filtered. The filtrate is then separated into solid and liquid again to obtain primary liquid fertilizer. S6: The primary liquid fertilizer is concentrated at ≤50℃ to a solid content of ≥40% to obtain rosemary fermented liquid fertilizer.

[0012] Furthermore, in step S4, the fermentation conditions are: temperature 28±1℃, fermentation time ≥10 days; spore suspension inoculation amount is at least 5% of the culture medium mass; solid-state aerobic fermentation, with turning at least once a day.

[0013] Furthermore, step S5 is as follows: after fermentation, water is added to the solid material for extraction; the amount of water added is at least 5 times the mass of the solid material, and the extraction is carried out for at least 4 hours; After extraction, filter the solution using a filter screen and collect the filtrate. Centrifuge the filtrate to separate the solid and liquid components, and take the supernatant to obtain the primary liquid fertilizer.

[0014] The second objective of this invention is to provide the application of the aforementioned organic water-soluble fertilizer in chili pepper cultivation.

[0015] Specifically, the organic water-soluble fertilizer is used in chili pepper cultivation to promote chili pepper growth and capsaicin synthesis, as well as to prevent chili pepper blight.

[0016] This invention provides a functional organic water-soluble fertilizer, which is a compound of marigold water-soluble fertilizer as the core nutrient supply carrier and rosemary fermentation liquid as the core antibacterial activity carrier. The two are scientifically formulated in a mass ratio of 2-3:1. Each component performs its own function, with no component antagonism and synergistic effect. The overall preparation process is designed around nutrient retention, antibacterial activity enrichment, and compliance control. The core principles are as follows: (a) The core function of marigold water-soluble fertilizer is to provide the necessary nutrients for crop growth in an all-round way.

[0017] This invention provides a marigold water-soluble fertilizer, which is a high-purity nutrient stock solution obtained from the marigold fermentation filtrate after solid-state fermentation pretreatment during lutein production. The filtrate is precisely adjusted to pH 6.0-6.5, boiled to remove foam, separated from solids, concentrated to a solid content of 50-60%, and then cooled and purified. Its core value lies in its single and efficient nutrient supply: the filtrate itself is rich in natural nitrogen, phosphorus, and potassium macroelements, as well as microelements such as calcium, magnesium, iron, and zinc from the marigold plant. It also contains humic acid, amino acids, and small molecule peptides, among other active organic nutrients. The refining process removes impurities and ineffective solid phases from the filtrate, retaining all water-soluble nutrients without any redundant components. It can directly provide the crops with the complete and essential nutrients required for growth and development, and is the core nutrient source of this water-soluble fertilizer.

[0018] The difficulty in preparing marigold water-soluble fertilizer lies in first adjusting the pH of the marigold fermentation filtrate to the range of 6.0–6.5. This is because when the pH is below 6.0, and the mass concentration of the marigold fermentation filtrate exceeds 50%, the concentrate will become a paste due to the precipitation of a large amount of potassium humate, making further concentration impossible. When the pH is above 6.5, the ammonium nitrogen in the marigold fermentation filtrate is easily lost as ammonia gas during the concentration process.

[0019] Secondly, during the concentration process of marigold fermentation filtrate, solids accounting for 1-2% of the original liquid mass will precipitate. The main components of these solids are inorganic salts such as marigold flower residue, potassium sulfate, calcium sulfate, and dicalcium phosphate. Some of these solids will form scale at the heating interface of the evaporator, thereby reducing the evaporation efficiency of the evaporator. If this problem is not solved, the concentration of marigold fermentation filtrate cannot continue. This invention has found that the main components of the scale are inorganic salts such as marigold flower residue, potassium sulfate, calcium sulfate, and dicalcium phosphate, which are easily soluble in inorganic acids such as hydrochloric acid and nitric acid. Therefore, this invention uses inorganic acids to clean the evaporator regularly (once every 24-72 hours). On the one hand, this allows the concentration of marigold fermentation filtrate to continue efficiently; on the other hand, the washing liquid can be returned to step S1 to participate in pH adjustment, without being discharged into the environment and polluting it.

[0020] (ii) The core function of rosemary fermentation liquid is to provide natural and highly effective antibacterial active substances in a targeted manner.

[0021] The rosemary fermentation broth in this invention is an active stock solution obtained from the solid waste residue of rosemary after essential oil extraction through aerobic fermentation, extraction, centrifugation, and low-temperature concentration. Its core value lies in the targeted supply of antibacterial substances. The fermentation broth contains a high concentration of live fungal spores and their antimicrobial metabolites secreted during fermentation (such as chitinase, protease, and antifungal peptides). After being applied to the soil (or sprayed on leaves or used for root irrigation), the bacteria can rapidly colonize the pepper roots, directly inhibiting and killing soil-borne pathogens such as Phytophthora capsici through nutrient competition, hyperparasitism, and the secretion of antimicrobial substances. The bacteria colonizing the roots also act as a "biological stimulant," continuously stimulating the pepper plant to activate its own systemic disease resistance, placing it in an "early warning" state, resulting in a faster and stronger response to subsequent pathogen invasion.

[0022] Rosemary waste contains natural antibacterial active ingredients such as rosmarinic acid, carrageenan, and ursolic acid. Through the bio-fermentation of bacteria, the large molecular components in the waste are degraded into small molecular active substances, while the metabolic products of the bacteria add new biocontrol and antibacterial components. The low-temperature concentration process (≤50℃) throughout the preparation process maximizes the preservation of the activity of all natural antibacterial active ingredients. This fermentation liquid contains no added chemical bactericides; its core function is to provide natural antibacterial substances, while only supplementing a small amount of organic nutrients. This is the core source of antibacterial and disease-preventing properties for this water-soluble fertilizer.

[0023] (III) Synergistic effect of compounding marigold water-soluble fertilizer and rosemary fermentation liquid The mass ratio of marigold water-soluble fertilizer to rosemary fermentation liquid is 2-3:1, which is a compliant ratio for nutrient supply and antibacterial protection. After this adjustment, it meets the standard requirements for macro-element type in "NY 1106-2010 Humic Acid Water-Soluble Fertilizer". It has good compatibility and can be used as a general-purpose fertilizer.

[0024] The marigold water-soluble fertilizer in this invention provides "food" for the bacteria: its rich soluble organic matter and nutrients provide an immediate source of energy and nutrition for the bacteria after they are applied to the soil, helping them to germinate, colonize and reproduce quickly, thereby consolidating and prolonging the effect of biological control.

[0025] The bacteria in rosemary fermentation liquid can enhance the efficacy of marigold components: the vigorous activity of bacteria in the rhizosphere can improve the rhizosphere microenvironment, promote the development of pepper roots, and thus improve the absorption and utilization efficiency of various biostimulants and nutrients in marigold fermentation liquid by peppers.

[0026] Capsaicin synthesis is a secondary metabolic reaction in chili peppers. The active ingredients of marigolds directly or indirectly participate in regulating its synthesis pathway, while mild stress induced by bacteria "activates" the defense metabolism of chili peppers. The two work synergistically to more effectively increase the capsaicin content in chili pepper fruits.

[0027] Phytophthora blight is a prevalent soil-borne fungal disease in chili pepper cultivation, caused by Phytophthora. It spreads rapidly and widely, causing significant yield losses once infected. Conventional chemical fungicides are prone to leaving pesticide residues and can lead to fungal resistance. However, the natural antibacterial substances such as rosmarinic acid and caryopsisic acid in rosemary fermentation liquid can target and inhibit the mycelial growth and spore germination of Phytophthora. Simultaneously, the fungal metabolites can colonize the chili pepper roots, forming a biological protective barrier and blocking the infection pathway. This antibacterial method combines biological inhibition and physical protection, resulting in significant control of Phytophthora blight and preventing its occurrence at its source. Furthermore, it leaves no pesticide residues, addressing the core pain points of difficult and persistent Phytophthora blight control.

[0028] In this invention, if marigold water-soluble fertilizer is missing: Rosemary fermentation liquid (microbial agent) will lose the crucial "nutrient support" it receives after being applied to the soil. Spores may colonize slowly and fail to establish a population due to intense competition in the soil and insufficient initial nutrients, resulting in unstable disease control and a shortened duration of effectiveness. Simultaneously, chili peppers will lose the comprehensive growth stimulation and potential capsaicin synthesis regulation provided by marigolds, leading to a significant reduction in growth-promoting and quality-improving effects. Ultimately, the product may degenerate into an ordinary biocontrol agent with unstable efficacy.

[0029] In this invention, if rosemary fermentation liquid is unavailable: The product will degenerate into a mere organic nutrient solution. Although marigold fermentation liquid can provide some growth-promoting effects, the plants will lose their active defense against soil-borne diseases (especially pepper blight). In areas with high incidence of blight or in fields with continuous cropping, the risk of pepper blight outbreaks will increase significantly. Simultaneously, due to the loss of induced systemic resistance, the overall health and stress resistance of pepper plants will decline, and the efficacy of the active ingredients in the marigold liquid cannot be maximized with the help of rhizosphere microorganisms, making it difficult to guarantee the effect of enhancing capsaicin synthesis.

[0030] The advantage of this invention lies in transforming two types of agricultural waste (marigold and rosemary residue) into high-value functional fertilizer, achieving resource recycling and integrated fertilizer and pesticide application. Through bio-fermentation, the marigold water-soluble fertilizer provides growth-promoting nutrients and enhances capsaicin synthesis, while the rosemary fermentation liquid introduces live bacteria, specifically targeting and controlling pepper blight. The two, when combined in a specific ratio, produce a synergistic effect: the bacteria receive nutrients, resulting in more stable preventative effects; the plants are protected, leading to superior growth. This significantly promotes pepper growth, improves fruit quality, and enhances disease resistance while reducing the use of chemical pesticides. Besides being particularly suitable for pepper cultivation, it is also a general-purpose water-soluble fertilizer. Detailed Implementation

[0031] To better understand the present invention, the present invention will be further described below with reference to specific serial numbers. The terminology used in the serial numbers is for describing specific embodiments and does not constitute a limitation on the scope of protection of the present invention.

[0032] In the specific implementation methods, unless otherwise specified, the experimental methods used are all conventional methods, and the materials and reagents used are all commercially available unless otherwise specified.

[0033] The bacteria used in this invention are those with accession number CICC13028, which were purchased from Mingzhou Biotechnology.

[0034] Example 1: Preparation of Marigold Water-Soluble Fertilizer Serial number (1) Raw materials used: Marigold fermentation filtrate: Marigold fermentation filtrate from a lutein factory; pH ​​3.5, containing: 11.1% solids, 3.5% amino acids, 1.67% K₂O, 0.57% total N, 0.38% ammonium nitrogen, and SO₄²⁻. 2- 1.88% Potassium carbonate (K2CO3): Prepared using commercially available quicklime, with a K2O content of over 35%.

[0035] Process steps: A. Adjust the pH of 4000g of marigold fermentation filtrate to 6.0 using potassium carbonate in a stirring tank; B. Add the marigold fermentation filtrate (with pH adjusted in step A) to a conventional evaporator and heat to boiling, while simultaneously removing organic foam. C. Filter the marigold fermentation filtrate heated in step B using a vacuum filter to obtain filtrate and filter residue. The filter residue is marigold flower residue that is returned to be dried, granulated and extracted for lutein extraction. D. The filtrate obtained in step C is sent to a conventional multi-effect evaporator to evaporate and concentrate to 840 grams, and the condensate during the evaporation process is collected. E. After cooling and settling, the concentrated liquid obtained in step D is sent to a settling tank to separate the solid and liquid. The organic matter in the solid is marigold residue that is returned to be dried, granulated and extracted to extract lutein. F. The liquid obtained in step E is sent to a conventional multi-effect evaporator for evaporation and concentration to 546 grams (containing 71.7% solids) to obtain the finished organic liquid fertilizer, whose effective components are: organic matter 41.0%, humic acid 29.0%, K2O 10.50%, total N 3.91%, ammonium nitrogen 2.42%, amino acids, etc. The condensate during the evaporation process is collected. G. Clean the evaporator in step D with industrial hydrochloric acid with a mass concentration of 3% every 8 hours, and return the cleaning solution to step A to participate in pH adjustment. The condensate from steps H, D, and F can be used as agricultural water.

[0036] Serial number (2) Raw materials used: Marigold fermentation filtrate: This is a fermentation filtrate of marigolds from a lutein factory, with a pH of 4.5. It contains: 11.1% solids, 1.67% K₂O, 0.57% total N, 0.38% ammonium nitrogen, and SO₄²⁻. 2- 1.88%; Ammonium bicarbonate: a commercially available product, with an nitrogen content of 17.0%; Process steps: A. Adjust the pH of 4000g of marigold fermentation filtrate to 6.2 using ammonium bicarbonate in a stirred tank; B. Add the marigold fermentation filtrate (with pH adjusted in step A) to a conventional evaporator and heat to boiling, while simultaneously removing organic foam. C. Filter the boiling liquid obtained in step B using a vacuum filter to obtain filtrate and filter residue. The filter residue is marigold flower residue, which is then dried, granulated, and used to extract lutein. D. The filtrate obtained in step C is sent to a conventional multi-effect evaporator and evaporated to 720 grams. The condensate during the evaporation process is collected. E. After cooling and settling, the concentrated liquid obtained in step D is sent to a settling tank to separate the liquid and solid. The organic matter in the solid is marigold residue that is returned to be dried, granulated and extracted to extract lutein. F. The liquid obtained in step E is sent to a conventional evaporator and evaporated to 542 grams (containing 71.8% solids) to obtain the finished organic water-soluble fertilizer. Its effective components are: organic matter 41.1%, humic acid 29.0%, K2O 9.71%, total N 4.90%, ammonium nitrogen 3.52%, and water-insoluble matter less than 5%. The condensate during the evaporation process is collected. G. Clean the evaporator from step D with 5% industrial salt or nitric acid every 72 hours, and return the cleaning solution to step A to participate in pH adjustment. The condensate from steps H, D, and F can be used as agricultural water.

[0037] Serial number (3) Raw materials used: Marigold fermentation filtrate: This is a fermentation filtrate of marigolds from a lutein factory, with a pH of 4.5. It contains: 11.1% solids, 1.67% K₂O, 0.57% total N, 0.38% ammonium nitrogen, and SO₄²⁻. 2- 1.88%; Potassium hydroxide: Commercially available, with a KOH content of 80.0%; Process steps: A. Adjust the pH of 4000g of marigold fermentation filtrate to 6.5 using potassium hydroxide in a stirring tank; B. The conditioning solution obtained in step A is sent to a conventional evaporator and heated to boiling, while simultaneously removing organic foam. C. Filter the solution obtained in step B using a vacuum filter to obtain filtrate and filter residue. The filter residue, namely marigold flower residue, is returned to be dried, granulated and extracted for lutein extraction. D. The filtrate obtained in step C is sent to a conventional falling film evaporator and evaporated to 780 grams. The condensate during the evaporation process is collected. E. After cooling and settling, the concentrated liquid obtained in step D is sent to a settling tank to separate the solid and liquid. The organic matter in the solid is marigold residue that is returned to be dried, granulated and extracted to extract lutein. F. The liquid obtained in step E is sent to a conventional falling film evaporator and evaporated to 551 grams (containing 71.8% solids) to obtain the finished organic liquid fertilizer. Its effective components are: organic matter 41.4%, humic acid 29.1%, K2O 12.50%, total N 3.90%, ammonium nitrogen 2.39%, amino acids, etc. The condensate during the evaporation process is collected. G. Clean the evaporator from step D with 4% industrial nitric acid every 14 hours, and return the cleaning solution to step A to participate in pH adjustment. The condensate from steps H, D, and F can be used as agricultural water.

[0038] In this embodiment, the marigold water-soluble fertilizers prepared by serial numbers (1), (2), and (3) all meet the relevant requirements of NY 1106-2010 for large element type liquid products in humic acid water-soluble fertilizers.

[0039] Example 2 Preparation of Rosemary Fermentation Broth Includes the following steps: S1: After extracting essential oil, the rosemary is dried at a temperature below 50°C until the moisture content is ≤15%; then it is crushed into particles with a particle size ≤2 mm to obtain rosemary waste residue; S2: Prepare the culture medium according to the following proportions by weight. Rosemary residue, 100 portions; Wheat bran, 10 parts; Peptone, 4 servings; Potassium dihydrogen phosphate, 0.2 parts; Magnesium sulfate heptahydrate, 0.1 parts; Adjust the water content of the culture medium to between 55-65%, and the initial pH to 5.0-5.5; S3: Take the preserved bacteria and incubate them in PDA medium at a constant temperature for 5-7 days until the surface of the medium is covered with mycelium and a large number of bluish-green spores begin to be produced.

[0040] Pour an appropriate amount of 0.05% Tween-80 sterile aqueous solution into a plate contaminated with spores. Gently scrape the surface of the culture medium with a sterile spreader or inoculation loop to fully release the spores into the liquid.

[0041] The harvested crude suspension was filtered, centrifuged and washed to collect the precipitate, and the suspension was adjusted to a concentration ≥1×10⁻⁶ with sterile water. 7 CFU / mL spore suspension; S4: Inoculate the spore suspension into the culture medium for fermentation under the following conditions: Temperature 28±1℃, fermentation time ≥10 days; spore suspension inoculation amount is 5% of the culture medium mass; solid-state aerobic fermentation, turning at least once a day.

[0042] The criteria for determining the completion of fermentation are: pH change ≤ 0.1 within 12 hours; the surface of the culture medium is completely covered with a green spore layer; the odor is a mild fermented aroma with a hint of earth mold, without any off-odors such as rancidity or ammonia.

[0043] S5: After fermentation, add 5 times the mass of water to the solid material; stir and extract at room temperature for 4 hours; After extraction, coarsely filter the solution using a filter screen to remove large solid residues; collect the filtrate. The filtrate was centrifuged at 8000 rpm for 10 min; the supernatant was collected. This yielded the primary liquid fertilizer. S6: The primary liquid fertilizer is concentrated at ≤50℃ to a solid content of ≥40% to obtain rosemary fermented liquid fertilizer.

[0044] Example 3: Preparation of Rosemary Inactivation Solution Includes the following steps: S1: After extracting essential oil, the rosemary is dried at a temperature below 50°C until the moisture content is ≤15%; then it is crushed into particles with a particle size ≤2 mm to obtain rosemary waste residue; S2: Prepare the culture medium according to the following proportions by weight. Rosemary residue, 100 portions; Wheat bran, 10 parts; Peptone, 4 servings; Potassium dihydrogen phosphate, 0.2 parts; Magnesium sulfate heptahydrate, 0.1 parts; Adjust the water content of the culture medium to between 55-65%, and the initial pH to 5.0-5.5; S3: Take the preserved bacteria and incubate them in PDA medium at a constant temperature for 5-7 days until the surface of the medium is covered with mycelium and a large number of bluish-green spores begin to be produced.

[0045] Pour an appropriate amount of 0.05% Tween-80 sterile aqueous solution into a plate contaminated with spores. Gently scrape the surface of the culture medium with a sterile spreader or inoculation loop to fully release the spores into the liquid.

[0046] The harvested crude suspension was filtered, centrifuged and washed to collect the precipitate, and the suspension was adjusted to a concentration ≥1×10⁻⁶ with sterile water. 7 CFU / mL spore suspension; S4: Inoculate the spore suspension into the culture medium for fermentation under the following conditions: Temperature 28±1℃, fermentation time ≥10 days; spore suspension inoculation amount is 5% of the culture medium mass; solid-state aerobic fermentation, turning at least once a day.

[0047] The criteria for determining the completion of fermentation are: pH change ≤ 0.1 within 12 hours; the surface of the culture medium is completely covered with a green spore layer; the odor is a mild fermented aroma with a hint of earth mold, without any off-odors such as rancidity or ammonia.

[0048] S5: After fermentation, sterilize at 121℃ for 15 minutes under high pressure; then add 5 times the mass of water to the solid material; stir and extract at room temperature for 4 hours. After extraction, coarsely filter the solution using a filter screen to remove large solid residues; collect the filtrate. The filtrate was centrifuged at 8000 rpm for 10 min; the supernatant was collected. This yielded the primary liquid fertilizer. S6: The primary liquid fertilizer is concentrated at ≤50℃ to a solid content of ≥40% to obtain rosemary inactivation solution.

[0049] Example 4 Preparation of Rosemary Extract Includes the following steps: S1: After extracting essential oil, the rosemary is dried at a temperature below 50°C until the moisture content is ≤15%; then it is crushed into particles with a particle size ≤2 mm to obtain rosemary waste residue; S2: Autoclave the rosemary residue at 121℃ for 10 min; then add 5 times its weight of water; stir and extract at room temperature for 4 h; After extraction, coarsely filter the solution using a filter screen to remove large solid residues; collect the filtrate. The filtrate was centrifuged at 8000 rpm for 10 min; the supernatant was collected. This yielded the primary liquid fertilizer. S3: The primary liquid fertilizer is concentrated at ≤50℃ to a solid content of ≥40% to obtain rosemary extract.

[0050] Example 5: Preparation of Organic Water-Soluble Fertilizer Weigh each raw material according to the mass fractions in Table 1; then stir evenly to obtain organic water-soluble fertilizer.

[0051] Table 1 Serial Number (1) Marigold Water-Soluble Fertilizer 2 0 0 3 3 0 1 2 2 Serial number (2) Marigold water-soluble fertilizer 0 2 0 0 0 0 0 0 0 Serial number (3) Marigold water-soluble fertilizer 0 0 2 0 0 0 0 0 0 Rosemary fermentation broth (Example 2) 1 1 1 1 0 3 1 0 0 Rosemary inactivation solution (Example 3) 0 0 0 0 0 0 0 1 0 Rosemary extract (Example 4) 0 0 0 0 0 0 0 0 1 Example 6 Antimicrobial Experiment of Phytophthora capsici I. Materials and Methods The tested strain was *Phytophthora capsici*.

[0052] II. Test Methods The Oxford cup method is used for measurement. Sterile Oxford cups are placed on agar plates inoculated with the target pathogen, and the fertilizer solution to be tested is added to the cups. Soluble chemicals and microbial metabolites in the fertilizer solution diffuse outwards through the contact surface between the bottom of the cup and the agar. If antibacterial components are present, a transparent inhibition zone forms around the Oxford cup. The cup walls physically prevent live bacteria in the fertilizer solution from entering the agar, thus solving the interference problem.

[0053] III. Experimental Procedure Instruments and consumables: sterile Oxford cups (6mm inner diameter, 8mm outer diameter, 10mm height), sterile petri dishes, constant temperature incubator, sterile pipettes and tips, vernier calipers, alcohol lamp, and laminar flow hood.

[0054] PDA culture medium is prepared and then sterilized for later use.

[0055] (1) Sample preparation: Water-soluble fertilizer solution containing live bacteria: The organic water-soluble fertilizers prepared in Example 5 were diluted 100 times with sterile water and then used for later use.

[0056] Preparation of reference standard: Positive control: Prepare 50 μg / mL nystatin for positive control.

[0057] Negative control: sterile water.

[0058] Target bacterial suspension: Prepare a fresh suspension of Phytophthora capsici spores and adjust the concentration to 1×10^6 CFU / mL.

[0059] (2) Preparation of bacterial plates Cool the sterilized culture medium to approximately 45-50°C.

[0060] In a clean bench, add 5 mL of target bacterial suspension for every 100 mL of culture medium and mix quickly.

[0061] Immediately pour the mixed culture medium containing bacteria into sterile petri dishes, about 15 mL per dish, and let it stand horizontally until it solidifies completely.

[0062] (3) Place the Oxford cup and add the sample On a solidified agar plate, use sterile forceps to pick up an Oxford cup, briefly flaming its outer wall over an alcohol lamp flame to sterilize it, and after cooling, gently place it vertically on the plate surface, applying slight pressure to ensure it is in close contact with the agar, preventing any gaps underneath that could lead to leakage. Place three Oxford cups evenly spaced on each plate.

[0063] Using a sterile pipette, carefully add equal volumes of different sample or control solutions to each Oxford cup.

[0064] After adding the sample, gently move the petri dish horizontally to ensure that the bottom of the Oxford cup is in complete contact with the agar and that there are no air bubbles.

[0065] (4) Cultivation and observation Place the plate upright in a constant temperature incubator and incubate at 25°C for 5 days.

[0066] Then measure the diameter of each inhibition zone.

[0067] The results are shown in Table 2.

[0068] Table 2 1 16.5 2 16.3 3 16.2 4 15.4 5 10.3 6 17.6 7 17.0 8 14.9 9 13.2 Positive control 19.3 negative control none The serial numbers in Table 2 follow the serial numbers of the organic water-soluble fertilizer prepared in Example 5.

[0069] Table 2 shows that items 1, 2, 3, and 4 all contain the chemical antibacterial components of marigold fertilizer and live bacteria and their metabolites from rosemary fermentation broth. The inhibition zones ranged from 15.4 mm to 16.5 mm, all significantly higher than item 5 (using marigold fertilizer alone), but lower than item 6 (using live bacteria fermentation broth alone). This indicates that: Limited synergistic effect: When marigold components are combined with live bacteria metabolites, a strong synergistic effect of "1+1>2" is not produced, and the total antibacterial efficacy is closer to the weighted average of the two.

[0070] At a fixed total volume, adding marigold fertilizer is equivalent to diluting the concentration of highly active live bacterial metabolites, which may result in the efficiency not reaching the level of pure fermentation broth. In addition, the pH or complex chemical composition of marigold fertilizer may slightly affect the stability or activity of live bacterial metabolites, which may be more obvious in item 4.

[0071] Differences in marigold fertilizer production processes have a negligible impact: Marigold fertilizers prepared using three different alkali sources were used in serial numbers 1, 2, and 3, but the differences in the inhibition zones (18.5, 18.3, 18.2 mm) were minimal, indicating that the chemical differences in the final product caused by different acid-base processes have no significant impact on the overall antibacterial efficacy after compounding.

[0072] Serial number 5 serves as a control for "containing only marigold water-soluble fertilizer (chemical components)". This data indicates that the basic chemical components (organic matter, humic acid, etc.) of the marigold fertilizer (process number 1) possess a certain antibacterial ability.

[0073] Serial number 6 serves as a control group containing only rosemary fermentation broth (live bacterial metabolites). It exhibits the largest inhibition zone, demonstrating that the live bacteria produced a large amount of highly efficient and diffusible antimicrobial metabolites during fermentation in the rosemary substrate.

[0074] Numbers 7, 8, and 9 use the same marigold fertilizer (process 1 in Example 1), but are combined with different states of rosemary components, demonstrating the key role of live bacteria: No. 7: Marigold fertilizer and rosemary fermentation broth (live bacteria). The inhibition zone was significantly larger than that of Nos. 1-4, and close to the highest level of No. 6. This verifies that when the proportion of live bacteria fermentation broth added is increased (1:1), its dominant role in the compound is enhanced.

[0075] No. 8: Marigold fertilizer and rosemary inactivation solution. The inhibition zone was significantly smaller than that of No. 7, which contained live bacteria. This indicates that while autoclaving kills live bacteria, it may also destroy some heat-sensitive antibacterial metabolites, leading to a decrease in antibacterial efficacy. However, residual heat-stable products and rosemary chemical extracts still contribute.

[0076] Serial No. 9: Marigold fertilizer and rosemary extract (unfermented). The inhibition zone is the smallest in this series. This demonstrates that unfermented rosemary raw material has limited antibacterial activity. The enhanced antibacterial ability is mainly attributed to the biotransformation by the bacteria, rather than simply the extraction of the raw material.

[0077] Example 7: Growth-promoting effect on chili seedlings The experiment was conducted using the Hangjiao No. 1 variety, and the method is as follows: First, the organic water-soluble fertilizer prepared in each serial number of Example 5 was mixed with commercial seedling substrate at a ratio of 1:50 (w / w) for pepper seedling cultivation. When the pepper seedlings grew to 4-6 true leaves, they were transplanted. Ten days after transplanting chili peppers, spray them once with organic water-soluble fertilizer diluted 100 times; the application rate is 50L / acre. In the early stage of fruit setting of peppers, apply organic water-soluble fertilizer (diluted 100 times) to the leaves, once every 10 days, for a total of 3 times, with an application rate of 50L / mu each time. Finally, 15 days before the chili peppers are harvested, spray the leaves once with a concentration of 25 mg / L abscisic acid, at a rate of 50 L / mu.

[0078] A pot experiment was conducted, and chili peppers were raised and transplanted according to the above experimental design. Twenty days after the chili pepper seedlings were transplanted, the plant height, stem diameter and capsaicin content were measured (all values ​​were averages of the samples). The results are shown in Table 3.

[0079] Commercially available organic fertilizers were used as a positive control group at the same dilution ratio.

[0080] Fertilizer-free cultivation was used as a negative control group in the experiment.

[0081] Table 3 1 46.78 6.24 0.371 2 46.57 6.26 0.364 3 46.37 6.24 0.369 4 48.54 6.32 0.354 5 47.04 6.30 0.323 6 41.18 5.45 0.335 7 43.74 5.78 0.357 8 46.23 6.18 0.346 9 44.11 5.89 0.331 Positive control 46.58 6.17 0.312 negative control 29.71 5.12 0.316 The serial numbers in Table 3 follow the serial numbers of the organic water-soluble fertilizer prepared in Example 5.

[0082] The data in Table 3 show that the plant height and stem diameter of all fertilization treatments were significantly higher than those of the negative control. This indicates that regardless of the formula, all treatments had a clear promoting effect on the growth of chili seedlings.

[0083] However, experimental data showed that the growth-promoting and antibacterial effects of the organic water-soluble fertilizers in each sequence of Example 5 did not exhibit a simple positive correlation, indicating that their mechanisms of action differed. The core function of rosemary fermentation broth (live bacteria) lies in its antibacterial effect: In the antibacterial experiment, the strongest performer (number 6, pure fermentation broth) exhibited the lowest plant height and stem diameter among all fertilization groups in the growth-promoting experiment, even lower than some other groups. This suggests that the main advantage of live bacteria and their metabolites may lie in secreting antibacterial substances, rather than directly providing nutrients or hormones that promote plant growth.

[0084] Marigold water-soluble fertilizer is a key foundation for promoting growth: Product number 5, containing only marigold water-soluble fertilizer, showed excellent performance in plant height and stem diameter, significantly better than the negative control and comparable to the positive control (commercially available organic fertilizer). This confirms that the organic matter, humic acid, potassium, nitrogen, and other nutrients contained in marigold water-soluble fertilizer are the key material basis for supporting the early growth of chili seedlings.

[0085] Most of the compound formulations (numbered 1-4, 7, 8) showed growth-promoting effects between pure marigold fertilizer and pure fermentation liquid in terms of plant height and stem diameter, and were generally equivalent to commercially available organic fertilizers (positive control), indicating that the compound formulations are effective overall.

[0086] Treatment number 4 exhibited the highest plant height and stem diameter among all treatments. Its formulation consisted of a 3:1 mixture of marigold fertilizer produced using a specific process and rosemary fermentation broth. This suggests that this particular ratio and process combination may have achieved an optimal balance in providing nutrients and regulating microbial activity / secondary metabolism, thus being most conducive to vegetative growth.

[0087] The growth promotion indicators (plant height 43.74 cm, stem diameter 5.78 mm) of No. 7 (a 1:1 mixture of marigold fertilizer and fermentation liquid) were relatively low, further supporting the observation that "a high proportion of fermentation liquid may not be entirely beneficial for early growth promotion," but its effect was still significantly better than the negative control.

[0088] The growth-promoting effect (plant height 46.23 cm, stem diameter 6.18 mm) of compound groups (combinations of compounds containing live bacteria fermentation liquid, such as compounds 1-3) was very similar to that of groups 1-3, and the capsaicin content (0.346 mg / g) was at a medium-to-high level. This indicates that sterilization treatment removes the potential competitive or inhibitory effects of live bacteria while retaining some beneficial metabolites produced during fermentation and substances from rosemary itself, which may be more conducive to the stable absorption of nutrients by plants, thus resulting in robust performance; however, it still lags behind the group containing live bacteria to some extent. The growth-promoting effect of No. 9 (marigold fertilizer + unfermented extract) was relatively weak; this once again confirms that rosemary raw materials that have not undergone microbial biotransformation provide limited growth-promoting substances or signaling substances directly.

[0089] Regarding the effect on capsaicin content, the measured capsaicin content in all fertilization treatment groups was higher than or close to the negative control (fertilizer-free cultivation) of 0.316 mg / g. This baseline indicates that fertilization treatments, as a whole, provide the material basis or inducing signal for capsaicin synthesis.

[0090] In the compound treatment containing live bacteria fermentation broth, the capsaicin content of samples 1, 2, and 3 (0.371, 0.364, and 0.369 mg / g, respectively) was the most outstanding, increasing by approximately 15% to 17% compared to the negative control. This indicates a synergistic effect between the comprehensive nutrition provided by the marigold water-soluble fertilizer and the specific metabolites or elicitors produced by the live bacteria, jointly promoting the biosynthesis of capsaicin.

[0091] Although the capsaicin content of sample 6 (pure rosemary fermentation liquid) was higher than that of the negative control, it was significantly lower than that of the compound group mentioned above. This indicates that, in the absence of sufficient basic nutrients provided by marigold fertilizer, the biostimulatory effect of live bacteria alone has a relatively limited effect on enhancing capsaicin synthesis.

[0092] The capsaicin content of sample number 5 (pure marigold water-soluble fertilizer) was slightly higher than that of the negative control. This confirms that the nutrients in the marigold fertilizer can support basic capsaicin synthesis. It is noteworthy that sample number 4, which showed the best growth-promoting effect (maximum plant height and stem diameter), had a capsaicin content of 0.354 mg / g, which, while higher than the average, was not the highest. This may suggest a certain resource allocation trade-off between rapid vegetative growth (plant height, stem diameter) and the accumulation of secondary metabolites (capsaicin).

[0093] The content of sample 8 (marigold fertilizer + inactivation solution) remained stable, indicating that some heat-stable substances produced during fermentation still have a positive impact on capsaicin synthesis. Sample 9 (marigold fertilizer + unfermented extract) showed a relatively weaker effect, further emphasizing the importance of the microbial biotransformation process for producing active substances that effectively stimulate capsaicin synthesis.

[0094] The above detailed description is a specific description of one of the feasible embodiments of the present invention. This embodiment is not intended to limit the patent scope of the present invention. All equivalent implementations or modifications that do not depart from the present invention should be included within the scope of the technical solution of the present invention.

Claims

1. An organic water-soluble fertilizer, characterized in that, Including marigold water-soluble fertilizer and rosemary fermentation liquid; the mass ratio of marigold water-soluble fertilizer to rosemary fermentation liquid is 2-3:1; The marigold water-soluble fertilizer is obtained by adjusting the pH of marigold fermentation filtrate, separating solid and liquid to obtain liquid, concentrating, cooling and settling to obtain liquid. The marigold fermentation filtrate is the liquid obtained by pressing and filtering after solid-state fermentation pretreatment when producing lutein using the marigold fermentation method. The rosemary fermentation broth was obtained by fermenting rosemary waste residue with bacteria of accession number CICC13028, followed by solid-liquid separation to obtain liquid. The rosemary residue is a solid waste product after essential oil extraction; The preparation of the marigold water-soluble fertilizer includes the following steps: S1: Adjust the pH of the marigold fermentation filtrate to 6.0-6.5; S2: Heat the marigold fermentation filtrate after pH adjustment to boiling and remove foam to obtain boiling liquid; S3: Separate the boiling liquid from the solid and liquid phases and collect the liquid phase; S4: The liquid phase is concentrated by evaporation to a solid content of 50-60%; a concentrated solution is obtained. S5: The concentrated liquid is cooled and settled to separate solid and liquid phases. The liquid phase is then taken as marigold water-soluble fertilizer. The preparation of the rosemary fermentation broth includes the following steps: A1: After extracting the essential oil, the rosemary is dried at a temperature below 50°C until the moisture content is ≤15%; then it is crushed into particles with a particle size ≤2 mm to obtain rosemary waste residue; A2: Prepare the culture medium according to the following proportions by weight. Rosemary residue, 100 portions; Carbon source, at least 10 parts; Nitrogen source, at least 4 parts; Potassium dihydrogen phosphate, at least 0.2 parts; Magnesium sulfate heptahydrate, at least 0.1 parts; Adjust the water content of the culture medium to between 55-65%, and the initial pH to 5.0-5.5; A3: Activate the bacteria with accession number CICC13028, and then prepare a concentration ≥1×10⁻⁶. 7 CFU / mL spore suspension; A4: Inoculate the spore suspension into the culture medium for aerobic fermentation; the criterion for determining the completion of fermentation is a pH change ≤0.1 within 12 hours; A5: After fermentation, water is added to the solid material for extraction, and then the filtrate is obtained by filtration; the filtrate is then separated into solid and liquid again to obtain primary liquid fertilizer; A6: Primary liquid fertilizer is concentrated at ≤50℃ to a solid content of ≥40% to obtain rosemary fermentation liquid; In step A4, the fermentation conditions are: temperature 28±1℃, fermentation time ≥10 days; spore suspension inoculum volume at least 5% of the culture medium mass; solid-state aerobic fermentation, with stirring at least once a day; step A5 is: After fermentation is complete, water is added to the solid material for extraction; the amount of water added is at least 5 times the mass of the solid material, and the extraction is carried out for at least 4 hours. After extraction, filter the solution using a filter screen and collect the filtrate. Centrifuge the filtrate to separate the solid and liquid components, and take the supernatant to obtain the primary liquid fertilizer.

2. The organic water-soluble fertilizer according to claim 1, characterized in that, The preparation of the marigold water-soluble fertilizer also includes the following steps: S6: Wash the solid phase separated in step S5, wherein the washing solution is returned to step S4; the washed solid phase is used to prepare lutein.

3. The organic water-soluble fertilizer according to claim 1, characterized in that, In step S4 of the preparation of the marigold water-soluble fertilizer, the evaporation container containing the liquid phase is cleaned with an acid washing agent, and the cleaning solution is returned to step S1; the acid washing agent is hydrochloric acid and / or nitric acid.

4. The application of the organic water-soluble fertilizer according to any one of claims 1-3 in chili pepper cultivation.

5. The application according to claim 4, characterized in that, The organic water-soluble fertilizer is used in chili pepper cultivation to promote chili pepper growth and capsaicin synthesis, as well as to prevent and control chili pepper diseases.