Functional proteoglycan organic fertilizer of lentinula edodes dregs, preparation method and application method thereof

Abstract

The present application relates to the technical field of agricultural fertilizer, and particularly relates to a functional proteoglycan organic fertilizer of lentinula edodes residue, a preparation method and an application method thereof.The organic fertilizer is composed of functional proteoglycan 40-60%, humic acid 15-30%, potassium dihydrogen phosphate 8-15%, biochar 5-10% and seaweed extract 5-10%, wherein the functional proteoglycan is prepared by mixing lentinula edodes residue, pleurotus ostreatus residue and agaric residue, is a proteoglycan with a molecular weight of 5-50 kDa and contains a beta-1,3 / 1,6-glucan structure; the preparation method comprises the steps of residue pretreatment, ultrasonic-assisted enzymatic hydrolysis, multi-stage membrane separation, concentration and drying, formula mixing, granulation and packaging and the like.The organic fertilizer has a dual function of promoting plant growth and activating the plant immune system, can significantly improve the disease resistance and yield of plants, and simultaneously improves the physical and chemical properties of soil; the preparation process is green and environmentally friendly, has high extraction efficiency and low energy consumption; and the pH value of the product is adjustable.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of agricultural fertilizer, in particular to a functional proteoglycan organic fertilizer of Lentinula edodes residue, a preparation method and an application method thereof. The organic fertilizer can provide nutrients required by plants, activate the immune system of plants, and improve the physical and chemical properties of soil, and is a multifunctional composite organic fertilizer. BACKGROUND

[0002] With the development of modern agriculture, improving crop yield and quality, improving soil environment, and reducing the use of chemical pesticides have become the main goal of current agricultural production. Traditional chemical fertilizers can provide nutrients required by crops, but long-term use will lead to a series of problems such as soil compaction, acidification, and microbial activity decline. Although organic fertilizer can improve soil structure, the nutrient release is slow, and the short-term fertilizer effect is not significant. Therefore, how to develop efficient organic fertilizer with nutrient supply and soil improvement functions has become a research hotspot in the field of agriculture.

[0003] The edible fungus industry is an important part of China's agriculture, with an annual output of more than 400 million tons, and also produces a large amount of waste mushroom residue. These residues are rich in organic matter, nitrogen, phosphorus, potassium and other nutrients and functional bioactive substances, and are ideal raw materials for preparing efficient organic fertilizer. However, the current utilization of edible fungus residue mainly focuses on direct composting or as a low-value cultivation medium, and the potential value of functional substances in the residue has not been fully tapped. As studies have shown, Lentinula edodes residue is usually treated by aerobic composting, and the fermentation efficiency and harmless treatment efficiency can be improved by adding other raw materials such as cow dung and composite microbial agents, but this treatment method still does not realize the high-value utilization of residue resources.

[0004] Chinese patent CN108794210A discloses an organic fertilizer containing Lentinula edodes residue and a preparation method thereof, which comprises the following raw materials by weight: 35-42 parts of Lentinula edodes residue, 12-19 parts of livestock and poultry manure, 5-9 parts of modified calcium carbonate, 10-14 parts of bone mud, 7-11 parts of filler, 6-9 parts of wood ash, 3-6 parts of sodium phosphate, 3-6 parts of potassium sulfate, 4-7 parts of calcium peroxide, 3-5 parts of sodium percarbonate, 4-8 parts of citric acid, 5-9 parts of sugar residue, 6-10 parts of composite fermentation bacteria, 3-5 parts of 35% ammonium hydroxide solution, 8-13 parts of urea, 8-11 parts of seaweed extract, and 12-16 parts of water.

[0005] The prior art has the following main problems: ① the utilization mode of the mushroom residue is single, mostly direct composting, and high-value utilization is not achieved; ② the extraction process of functional substances is low in efficiency, high in energy consumption and cost; ③ the prepared products are single in function, most of which only have the effect of conventional fertilizer and lack special functions such as plant immune activation; ④ the applicability is limited, and the adaptability to different soil types is poor. In addition, in modern agriculture, plant growth-promoting bacteria can significantly improve soil quality, increase soil fertility, improve crop stress resistance and increase yield and quality, but plant immune activation and organic fertilizer have not been effectively combined.

[0006] Therefore, it is urgent to develop an organic fertilizer capable of efficiently utilizing edible mushroom waste, extracting functional proteoglycans therefrom, having dual functions of nutrient supply and plant immune activation, and realizing a low-energy-consumption and green environmental protection preparation process. SUMMARY

[0007] The purpose of the application is to provide a Lentinula edodes residue functional proteoglycan organic fertilizer, a preparation method and an application method thereof, aiming to solve the technical problems of low utilization rate, single function and high energy consumption of the preparation process of the existing mushroom residue, realize high-value utilization of the mushroom residue resource, and provide a multifunctional organic fertilizer with growth promotion and immune activation functions.

[0008] To achieve the above-mentioned purpose, the application provides the following technical solutions:

[0009] The Lentinula edodes residue functional proteoglycan organic fertilizer is composed of the following components by weight percentage:

[0010] functional proteoglycan 40-60%,

[0011] humic acid 15-30%,

[0012] potassium dihydrogen phosphate 8-15%,

[0013] biochar 5-10%,

[0014] seaweed extract 5-10%;

[0015] The functional proteoglycan is a proteoglycan with a molecular weight of 5-50 kDa and contains a β-1,3 / 1,6-glucan structure.

[0016] As a preferred, the functional proteoglycan is prepared from the following weight ratio of residue mixture: Lentinula edodes residue 50-70%, Pleurotus ostreatus residue 15-25%, and Auricularia auricula residue 15-25%.

[0017] As a preferred, the physicochemical properties of the organic fertilizer are:

[0018] pH value 5.0-9.0,

[0019] Organic matter content ≥ 45%,

[0020] Total nutrient N + P2O5+ K2O content ≥ 8%,

[0021] Moisture content ≤ 15%,

[0022] Particle size 1-5 mm.

[0023] As preferred, the humic acid is rich in carboxyl and phenolic hydroxyl groups, with strong complexing ability; the specific surface area of the biochar is ≥ 300 m² / g, and the porosity is ≥ 60%; the seaweed extract contains ≥ 20% alginic acid and ≥ 5% mannitol.

[0024] The method for preparing the functional proteoglycan organic fertilizer from shiitake mushroom residue comprises the following steps:

[0025] (1) Residue pretreatment: mix shiitake mushroom residue, pleurotus ostreatus residue, and agaric residue, crush to a particle size ≤ 0.5 mm, add 0.3-0.8% NaOH solution, control the pH value to 8.0-9.5, and pretreat at 55-65°C for 1.5-2.5 hours;

[0026] (2) Ultrasonic-assisted enzymatic hydrolysis: add a complex enzyme system to the pretreated mixed residue, the complex enzyme system comprising cellulase, hemicellulase, and protease, the enzyme addition amount being 1-3% of the substrate mass, adjust the pH value to 5.5-6.0, control the temperature at 45-50°C, apply ultrasonic treatment at a power of 400-600 W and a frequency of 35-45 kHz, and hydrolyze for 4-6 hours;

[0027] (3) Multistage membrane separation: after solid-liquid separation of the enzymatic hydrolysate, sequentially pass the supernatant through membrane separation devices with a 100 kDa, 50 kDa, and 5 kDa molecular weight cut-off, and collect the proteoglycan component with a molecular weight of 5-50 kDa;

[0028] (4) Concentration and drying: concentrate the collected proteoglycan solution to a concentration of 15-25%, and use a spray drying process to prepare proteoglycan powder at an inlet temperature of 170-190°C and an outlet temperature of 75-85°C;

[0029] (5) Formula mixing: uniformly mix the proteoglycan powder with humic acid, potassium dihydrogen phosphate, biochar, and seaweed extract according to the weight percentages;

[0030] (6) Granulation and packaging: pass the mixed material through an extrusion granulation device to prepare granules with a particle size of 1-5 mm, dry to a moisture content of ≤ 15%, cool, sieve, and then package.

[0031] As preferred, in step (1), the pretreatment is performed in a reaction kettle with a stirring speed of 100-200 rpm, and the solid-liquid ratio is 1:5-1:8.

[0032] As preferred, in the step (2), the ultrasonic treatment is in an intermittent mode, with 2-3 minutes of opening and 1-2 minutes of pause, and the total ultrasonic treatment time is 30-45 minutes.

[0033] As preferred, in the step (3), the membrane separation operation pressure is 0.2-0.4 MPa, the temperature is 25-30℃, and the pH value is controlled at 6.0-7.0.

[0034] As preferred, in the step (5), the mixing is performed by using a V-shaped mixer, the mixing time is 15-30 minutes, the rotating speed is 15-25 rpm, the proteoglycan powder is mixed with the humic acid first, then the potassium dihydrogen phosphate is added, and finally the biochar and the seaweed extract are added.

[0035] The application method of the functional proteoglycan organic fertilizer of the Lentinula edodes residue comprises the following steps:

[0036] (1) Basic application: 7-15 days before the sowing or planting of crops, the organic fertilizer is uniformly applied to the plough layer and mixed with the soil by ploughing, and the application amount is 30-50 kg / mu;

[0037] (2) Additional application: in the key period of crop growth, the water and fertilizer are integrated by using the irrigation system, and the organic fertilizer is used by being diluted by 500-1000 times;

[0038] (3) Foliar spraying: 7-10 days before the high incidence period of diseases, the organic fertilizer is diluted by 1000-2000 times, and is uniformly sprayed on the plant leaves, once every 7-15 days, for 2-3 times continuously;

[0039] It is suitable for various soil types with a pH value of 5.0-9.0, and is suitable for crops including vegetables, fruit trees and economic crops.

[0040] Compared with the prior art, the present application has the following beneficial effects:

[0041] 1. The edible fungus waste residue is used for high-value utilization, the resource recycling is realized, the extracted functional proteoglycan can significantly improve the plant disease resistance, the disease resistance of tomatoes is increased by 50%, and the yield is increased by 30%;

[0042] 2. The green process combining ultrasonic-assisted enzymolysis and multi-stage membrane separation is innovatively adopted, the extraction efficiency is increased by 80% than that of the traditional acidolysis method, no waste liquid is discharged, and the energy consumption is reduced by 40%;

[0043] 3. Through the synergistic design of the formula components, the product has multiple functions of nutrient supply, plant immune activation and soil improvement, and the pH value is adjustable (5.0-9.0), which is suitable for various soil types such as acid soil, neutral soil and alkaline soil;

[0044] 4. Application mode is diversified, can be base application, post application and foliar spraying, suitable for vegetables, fruit trees and economic crops and various crops, and has wide application prospect. DETAILED DESCRIPTION

[0045] The application will be further described in detail through specific examples, but the protection scope of the application is not limited thereto. Unless otherwise specified, the technical methods and materials used in the application are methods and materials known to those skilled in the art. The application will be further described in detail through specific examples, but the protection scope of the application is not limited to the following examples.

[0046] The raw materials used in the application are all commercially available or obtained by conventional methods. Unless otherwise specified, the methods used in the examples of the application are conventional methods.

[0047] The key of the application lies in efficient extraction of functional proteoglycans from edible mushroom waste. Proteoglycans are a class of glycoprotein complexes composed of protein skeleton and covalently linked sugar chains, which play an important role as signal molecules in plants, can activate the plant immune system and improve plant stress resistance. Edible mushroom waste contains rich proteoglycans, mainly including β-1, 3 / 1, 6-glucan structure, which is recognized by plants as pathogen-associated molecular patterns (PAMPs) and can induce plant systemic acquired resistance (SAR), thereby activating the plant immune system.

[0048] Through research, it is found that the content and structure of proteoglycans in different edible mushroom waste are different, therefore, the application adopts the mixed extraction method of shiitake mushroom waste, shiitake mushroom waste and agaric waste to obtain a mixture of proteoglycans with diverse structures and complementary functions. Among them, shiitake mushroom waste mainly contains proteoglycans with large molecular weight, high β-1, 3 glycosidic bond content; shiitake mushroom waste contains proteoglycans with moderate molecular weight and high β-1, 6 glycosidic bond content; agaric waste contains special sulfated proteoglycans with strong plant immune activity. By reasonably proportioning the three kinds of waste, a proteoglycan product with more complete structure and more diverse functions can be obtained.

[0049] In the extraction process, the application innovatively adopts a green process combining ultrasonic-assisted enzymatic hydrolysis and multi-stage membrane separation. Ultrasonic treatment can destroy the cell wall structure of the waste and increase the enzymatic hydrolysis efficiency; multi-stage membrane separation can accurately separate the proteoglycans according to their molecular weight, and obtain products with higher activity of specific molecular weight. Compared with the traditional acid hydrolysis method, the process of the application not only improves the extraction efficiency, but also avoids the generation of acid and alkali waste liquid, realizing green and environmental protection.

[0050] Example 1: Shiitake mushroom waste functional proteoglycan organic fertilizer with standard formula

[0051] The organic fertilizer of the present embodiment is composed of the following components in percentage by weight: functional proteoglycan 50%, humic acid 25%, potassium dihydrogen phosphate 10%, biochar 8%, and seaweed extract 7%.

[0052] The functional proteoglycan is prepared from the following mixture of mushroom residues in weight ratio: Lentinula edodes residue 60%, Pleurotus ostreatus residue 20%, and Auricularia auricula residue 20%.

[0053] The humic acid is prepared by using weathered coal as the main raw material and combining with a microbial fermentation process, and the specific steps are as follows:

[0054] (1) Raw material screening and pretreatment: high-quality weathered lignite with humic substance content ≥ 60% is selected as the raw material, and after removing obvious impurities, it is crushed to a particle size of 0.2-0.3 mm, and then the crushed material is dried at 80°C to a water content of less than 5%;

[0055] (2) Pre-oxidation treatment: the dried coal powder is mixed with 3% hydrogen peroxide solution (solid-liquid ratio of 1:5), stirred (150 rpm) at 40°C for 4 hours, so that the organic matter structure in the coal is partially oxidized, and the subsequent alkali extraction efficiency is increased;

[0056] (3) Alkali extraction: the pre-oxidized material is added to 5% NaOH solution (solid-liquid ratio of 1:6), and extracted by reflux at 85°C for 3 hours with a stirring speed of 180 rpm; during the extraction process, the humic acid leaching rate is measured every 30 minutes to ensure sufficient extraction;

[0057] (4) Solid-liquid separation: the extraction liquid is filtered through a 120-mesh screen, and the filter residue is returned to step (3) for secondary extraction, and the two filtrates are combined;

[0058] (5) Acid precipitation: the combined filtrate is adjusted to a pH of 2.0 with concentrated sulfuric acid, slowly stirred (60 rpm), and left to stand at room temperature for 8 hours to allow the humic acid to fully precipitate;

[0059] (6) Humic acid separation: the precipitated humic acid is collected by centrifugal separation (4000 rpm, 25 minutes), and the precipitate is washed with deionized water for 3 times to remove residual inorganic salts;

[0060] (7) Neutralization and conditioning: the washed humic acid is mixed with 2% KOH solution to adjust the pH to 7.0-7.5 to form water-soluble potassium humate, and after uniform stirring (120 rpm), it is left to stand for 2 hours;

[0061] (8) Microbial fermentation strengthening: humic substance degrading bacteria (Bacillus subtilis, Pseudomonas, and Actinomyces, in a ratio of 2:2:1, with a total bacterial amount ≥ 2×10 8The fermentation was carried out at 30°C for 48 hours with 5% inoculation amount, and intermittent stirring (10 minutes every 2 hours) and micro-aeration (dissolved oxygen 2-4 mg / L) were maintained during the fermentation;

[0062] (9) Functional modification: after the fermentation, 2% amino acids (glycine and alanine, mass ratio 3:2) were added to the solution, and the reaction was carried out at 50°C for 4 hours to enhance the chelating ability of humic acid to trace elements;

[0063] (10) Membrane concentration: the functionalized humic acid solution was concentrated to 1 / 4 of the original volume using an ultrafiltration membrane (molecular weight cut-off 3 kDa);

[0064] (11) Spray drying: the concentrated solution was made into a powdered humic acid product using spray drying technology (inlet temperature 165°C, outlet temperature 70°C, atomization pressure 2.5 MPa);

[0065] (12) Quality testing: the final product was tested to have humic acid content ≥75%, pH value 6.8-7.2, water solubility ≥90%, organic matter content ≥65%, potassium content ≥8%, rich in carboxyl groups (≥3.8 mmol / g) and phenolic hydroxyl groups (≥2.5 mmol / g), and strong ion exchange capacity and complexing ability.

[0066] The humic acid in this embodiment is prepared by extracting weathered coal combined with microbial fermentation strengthening process, and has the characteristics of high activity, high water solubility, and strong chelating ability. The rich carboxyl and phenolic hydroxyl structures can effectively improve the soil aggregate structure, promote soil microbial activity, and enhance the soil's ability to retain fertilizer and water; at the same time, they can also synergize with proteoglycans to promote nutrient uptake by plants and enhance plant disease resistance. The microbial fermentation strengthening process allows the final product to contain a variety of beneficial microbial metabolites, further improving the biological activity and fertilizer efficiency persistence of humic acid.

[0067] Detailed preparation method of biochar:

[0068] Biochar is prepared using agricultural waste (rice husk) as raw material, and the specific steps are as follows:

[0069] (1) Raw material pretreatment: collect rice husk, wash and dry at 80°C to a moisture content of less than 10%, then crush to 3-5 mm in size;

[0070] (2) High-temperature pyrolysis: put the pretreated rice husk into a rotary kiln type pyrolysis furnace with controllable internal temperature, slowly heat to 350°C under nitrogen protection, maintain for 1 hour; then heat to 500°C, maintain for 2 hours for complete carbonization. The heating rate is controlled at 10°C / min during pyrolysis to ensure uniform carbonization;

[0071] (3) Activation treatment: After the primary carbonization of the biochar, it is cooled to room temperature and then immersed in a 1.5 mol / L phosphoric acid solution with a liquid-solid ratio of 4:1 for 12 hours. Subsequent pyrolysis is performed at 650°C under a nitrogen atmosphere for 1.5 hours to increase the porosity and specific surface area of the biochar;

[0072] (4) Post-treatment: The activated biochar is repeatedly washed with deionized water until it is neutral, dried at 105°C for 12 hours, and then sieved through a 180-mesh screen to obtain high-quality biochar with uniform particles, a specific surface area of ≥350 m² / g, and a porosity of about 65%;

[0073] (5) Functionalization treatment: To improve the adsorption and slow-release capacity of the biochar for nutrient elements, it is mixed with a 0.5% chitosan solution (mass ratio of 10:1), stirred uniformly, and then dried at 60°C for 4 hours to obtain modified biochar with good adsorption capacity and slow-release function.

[0074] Detailed preparation method of seaweed extract:

[0075] The seaweed extract is prepared using brown algae (a mixture of kelp and sargassum) as raw material, and the specific steps are as follows:

[0076] (1) Raw material preparation: Fresh brown algae (kelp and sargassum mixed in a mass ratio of 1:1) is selected, washed, and cut into small pieces, then dried at 60°C until the water content is less than 15%, and then pulverized to a particle size of less than 2mm;

[0077] (2) Alkaline extraction: Dry brown algae powder is added to a 0.8% NaOH solution (solid-liquid ratio of 1:15), and extracted at 60°C for 3 hours, with stirring (150 rpm) during the extraction process;

[0078] (3) Acid precipitation: The extract is filtered through a 100-mesh screen, and the pH is adjusted to 3.0 with hydrochloric acid. It is then allowed to stand at room temperature for 4 hours to precipitate the alginic acid;

[0079] (4) Alcohol extraction: The treated solution is added to 3 times the volume of 95% ethanol, and allowed to stand at room temperature for 8 hours to precipitate the components rich in fucoidan and mannitol;

[0080] (5) Separation and purification: The precipitate is collected by centrifugation (5000 rpm, 20 minutes), dissolved in deionized water, treated with activated carbon for 2 hours to decolorize, and then filtered through a 0.45μm filter membrane;

[0081] (6) Concentration and drying: The purified solution is concentrated to 1 / 5 of the original volume under reduced pressure (-0.08 MPa), and then spray-dried (inlet temperature 160°C, outlet temperature 70°C) to produce powdered seaweed extract.

[0082] (7)Quality detection: The content of alginate in the final product is ≥25%, the content of mannitol is ≥8%, and the content of fucoidan is ≥10%, which has strong plant growth regulation and immune activation functions.

[0083] Preparation method of functional proteoglycan organic fertilizer of shiitake mushroom residue

[0084] (1) Residue pretreatment: mix shiitake mushroom residue, shiitake mushroom residue and agaric residue according to the weight ratio of 60:20:20, crush to particle size 0.4mm, add 0.5% NaOH solution (solid-liquid ratio 1:6), control pH value 9.0, pretreat at 60℃ for 2 hours, stirring speed 150rpm;

[0085] (2) Ultrasonic-assisted enzymolysis: add compound enzyme system (cellulase, hemicellulase and protease mass ratio 2:2:1) to the pretreated mixed residue, enzyme addition amount is 2% of the substrate mass, adjust pH value to 5.8, control temperature at 48℃, apply ultrasonic treatment (power 500W, frequency 40kHz) at the same time, adopt intermittent mode (open for 2.5 minutes, pause for 1.5 minutes), total ultrasonic treatment time 40 minutes, enzymolysis time 5 hours;

[0086] (3) Multistage membrane separation: centrifuge the enzymolysis liquid (5000rpm, 15 minutes), take the supernatant and pass through hollow fiber membranes with molecular weight cut-off of 100kDa, 50kDa and 5kDa in turn, operating pressure 0.3MPa, temperature 28℃, pH value 6.5, collect the proteoglycan components with molecular weight of 5-50kDa;

[0087] (4) Concentration and drying: concentrate the collected proteoglycan solution to 20% concentration with a rotary evaporator, and then use spray drying process (inlet temperature 180℃, outlet temperature 80℃, feeding rate 20L / h) to make proteoglycan powder;

[0088] (5) Formula mixing: put 50kg of proteoglycan powder, 25kg of humic acid, 10kg of potassium dihydrogen phosphate, 8kg of the prepared biochar and 7kg of the prepared seaweed extract into a V-type mixer, mix for 25 minutes at a speed of 20rpm, first mix the proteoglycan powder with the humic acid, then add the potassium dihydrogen phosphate, and finally add the biochar and seaweed extract;

[0089] (6) Granulation and packaging: make the mixture into granules by extrusion granulation equipment (pore size 3mm), dry at 60℃ for 6 hours to a moisture content of 12%, cool to room temperature, then sieve, collect the granules with size of 1-5mm, and package into plastic woven bags, 25kg per bag.

[0090] By the above detailed preparation method of biochar and seaweed extract, the quality and stability of active components in the final organic fertilizer product are ensured, making the product have excellent performance in plant immune activation and soil improvement. The high specific surface area and pore structure of biochar are beneficial to improve soil structure and water and fertilizer retention performance, while the active substances such as alginate, mannitol and fucoidan in seaweed extract can effectively promote plant growth and enhance plant resistance and immunity. Proteoglycans and humic acid synergistically enhance plant root development and nutrient absorption. Biochar provides a habitat for microorganisms, enhancing microbial activity and persistence; the growth-regulating substances in seaweed extract and the immune-activating function of proteoglycans complement each other; potassium dihydrogen phosphate provides available nutrients to support plant growth in the early stage and enhance the immune-activating effect of proteoglycans.

[0091] Product characteristics:

[0092] Appearance: brown granular, no odor

[0093] pH: 6.8

[0094] Organic matter content: 52.3%

[0095] Total nutrient (N+P2O5+K2O) content: 9.2%

[0096] Moisture content: 12%

[0097] Particle size: 1-5 mm.

[0098] Example 2: Organic fertilizer with high proteoglycan content formula

[0099] The organic fertilizer of this example is composed of the following components by weight percentage: functional proteoglycan 60%, humic acid 20%, potassium dihydrogen phosphate 8%, biochar 6%, and seaweed extract 6%.

[0100] Among them, the functional proteoglycan is prepared from the following weight ratio of mushroom residue mixture: shiitake mushroom residue 70%, shiitake mushroom residue 15%, and agaric mushroom residue 15%.

[0101] Preparation method:

[0102] (1) Residue pretreatment: Mix shiitake mushroom residue, shiitake mushroom residue and agaric mushroom residue at a weight ratio of 70:15:15, crush to a particle size of 0.3 mm, add 0.7% NaOH solution (solid-liquid ratio 1:5), control pH 9.2, pretreat at 62°C for 2.2 hours, stirring speed 180 rpm;

[0103] (2) Ultrasonic-assisted enzymatic hydrolysis: A complex enzyme system (cellulase, hemicellulase, and protease with a mass ratio of 2.5:2:1) was added to the pretreated mixed microbial residue. The enzyme addition amount was 2.5% of the substrate mass, the pH value was adjusted to 5.6, the temperature was controlled at 49°C, and ultrasonic treatment (power 550W, frequency 42kHz) was applied. The intermittent mode (on for 2 minutes, off for 1 minute) was adopted, and the total ultrasonic treatment time was 45 minutes. The enzymatic hydrolysis time was 5.5 hours;

[0104] (3) Multistage membrane separation: The enzyme hydrolysate was centrifuged (5500 rpm, 20 minutes), and the supernatant was sequentially passed through hollow fiber membranes with a molecular weight cutoff of 100kDa, 50kDa, and 5kDa. The operating pressure was 0.35MPa, the temperature was 26°C, the pH value was 6.8, and the 5-50kDa proteoglycan component was collected;

[0105] (4) Concentration and drying: The collected proteoglycan solution was concentrated to 25% concentration using a rotary evaporator, and then spray-dried (inlet temperature 185°C, outlet temperature 85°C, feed rate 18L / h) to produce proteoglycan powder;

[0106] (5) Formula mixing: 60kg of proteoglycan powder, 20kg of humic acid, 8kg of potassium dihydrogen phosphate, 6kg of biochar, and 6kg of seaweed extract were added to a double-screw mixer. The mixing time was 20 minutes, the rotation speed was 25rpm, and the mixing sequence was the same as in Example 1.

[0107] (6) Granulation and packaging: The mixed material was granulated by disc granulation equipment, dried at 65°C for 5 hours to a moisture content of 10%, cooled to room temperature, sieved, and the 1-4mm granules were collected and packaged into composite plastic bags, each bag containing 20kg.

[0108] Product characteristics:

[0109] Appearance: dark brown granules, no odor

[0110] pH value: 7.2

[0111] Organic matter content: 56.8%

[0112] Total nutrient (N+P2O5+K2O) content: 8.6%

[0113] Moisture content: 10%

[0114] Particle size: 1-4mm.

[0115] Example 3: High humic acid formula organic fertilizer

[0116] The organic fertilizer of the present embodiment is composed of the following components in percentage by weight: functional proteoglycan 45%, humic acid 30%, potassium dihydrogen phosphate 10%, biochar 7%, seaweed extract 8%.

[0117] The functional proteoglycan is prepared from the following mixture of mushroom residues in weight ratio: Lentinula edodes mushroom residue 55%, Pleurotus ostreatus mushroom residue 25%, Auricularia auricular mushroom residue 20%.

[0118] Preparation method:

[0119] (1) Residue pretreatment: mix Lentinula edodes mushroom residue, Pleurotus ostreatus mushroom residue and Auricularia auricular mushroom residue in weight ratio of 55:25:20, crush to particle size of 0.45 mm, add 0.4% NaOH solution (solid-liquid ratio 1:7), control pH value to 8.5, pretreat at 58°C for 1.8 hours, stirring speed 130 rpm;

[0120] (2) Ultrasonic-assisted enzymatic hydrolysis: add a complex enzyme system (cellulase, hemicellulase and protease in mass ratio of 1.5:2:1.5) to the pretreated mixed mushroom residue, enzyme addition amount is 1.8% of the substrate mass, adjust pH value to 5.7, control temperature at 47°C, apply ultrasonic treatment (power 450 W, frequency 38 kHz) at the same time, adopt intermittent mode (turn on for 3 minutes, pause for 1.5 minutes), total ultrasonic treatment time 35 minutes, enzymatic hydrolysis time 4.5 hours;

[0121] (3) Multistage membrane separation: centrifuge the enzymatic hydrolysate (4800 rpm, 18 minutes), take the supernatant and pass through hollow fiber membranes with molecular weight cut-off of 100 kDa, 50 kDa and 5 kDa in sequence, operating pressure 0.25 MPa, temperature 27°C, pH value 6.3, collect the proteoglycan component of 5-50 kDa;

[0122] (4) Concentration and drying: concentrate the collected proteoglycan solution to 18% concentration with a rotary evaporator, then use spray drying process (inlet temperature 175°C, outlet temperature 78°C, feed rate 22 L / h) to make proteoglycan powder;

[0123] (5) Formula mixing: put 45 kg of proteoglycan powder, 30 kg of humic acid, 10 kg of potassium dihydrogen phosphate, 7 kg of biochar and 8 kg of seaweed extract into a double-cone mixer, mix for 30 minutes at a speed of 15 rpm, and the mixing sequence is the same as in Example 1;

[0124] (6) Granulation and packaging: pass the mixed material through an extrusion granulation device (pore size 2.5 mm) to make granules, dry at 62°C for 5.5 hours to a moisture content of 13%, cool to room temperature, sieve and collect granules of 1-4.5 mm, and pack into kraft paper bags, 15 kg per bag.

[0125] Product characteristics:

[0126] Appearance: black-brown granules, no odor

[0127] pH: 6.5

[0128] Organic matter content: 58.5%

[0129] Total nutrient (N+P2O5+K2O) content: 9.8%

[0130] Moisture content: 13%

[0131] Particle size: 1-4.5 mm.

[0132] Example 4: Organic fertilizer with high phosphorus formula

[0133] The organic fertilizer of this example is composed of the following components in weight percentage: functional proteoglycan 40%, humic acid 25%, monopotassium phosphate 15%, biochar 10%, and seaweed extract 10%.

[0134] Among them, the functional proteoglycan is prepared from the following weight ratio of fungus residue mixture: shiitake mushroom residue 50%, oyster mushroom residue 25%, and agaric fungus residue 25%.

[0135] Preparation method:

[0136] (1) Fungus residue pretreatment: mix shiitake mushroom residue, oyster mushroom residue, and agaric fungus residue in a weight ratio of 50:25:25, crush to a particle size of 0.5 mm, add 0.3% NaOH solution (solid-liquid ratio 1:8), control pH value to 8.2, pretreat at 55°C for 1.5 hours, stirring speed 120 rpm;

[0137] (2) Ultrasonic-assisted enzymatic hydrolysis: add a complex enzyme system (cellulase, hemicellulase, and protease in a mass ratio of 1:2:1) to the pretreated mixed fungus residue, enzyme addition amount is 1.5% of the substrate mass, adjust pH value to 5.5, control temperature at 45°C, and apply ultrasonic treatment (power 400W, frequency 35kHz) at the same time, adopt intermittent mode (turn on for 2 minutes, pause for 2 minutes), total ultrasonic treatment time 30 minutes, enzymatic hydrolysis time 4 hours;

[0138] (3) Multistage membrane separation: centrifuge the enzymatic hydrolysate (4500 rpm, 15 minutes), take the supernatant and pass through hollow fiber membranes with molecular weight cut-off of 100kDa, 50kDa, and 5kDa in sequence, operating pressure 0.2MPa, temperature 25°C, pH value 6.0, collect the proteoglycan component with molecular weight of 5-50kDa;

[0139] (4) Concentration and drying: The collected proteoglycan solution was concentrated to 15% concentration by using a rotary evaporator, and then a spray drying process was used to make proteoglycan powder (inlet temperature 170°C, outlet temperature 75°C, feeding rate 25L / h);

[0140] (5) Formula mixing: 40 kg of proteoglycan powder, 25 kg of humic acid, 15 kg of potassium dihydrogen phosphate, 10 kg of biochar, and 10 kg of seaweed extract were put into a V-type mixer, mixed for 30 minutes at a speed of 18 rpm, and the mixing sequence was the same as in Example 1.

[0141] (6) Granulation and packaging: The mixed material was made into granules by a pressure roller granulation device, dried at 58°C for 7 hours to a moisture content of 11%, cooled to room temperature, and then sieved to collect 1.5-5mm granules, which were packaged into woven bags, each containing 25 kg.

[0142] Product characteristics:

[0143] Appearance: Brown granules, no odor

[0144] pH: 6.3

[0145] Organic matter content: 49.6%

[0146] Total nutrient (N+P2O5+K2O) content: 12.5%

[0147] Moisture content: 11%

[0148] Particle size: 1.5-5mm.

[0149] Example 5: Organic fertilizer suitable for acidic soil

[0150] The organic fertilizer of this example is composed of the following components by weight percentage: functional proteoglycan 45%, humic acid 20%, potassium dihydrogen phosphate 10%, biochar 15%, and seaweed extract 10%.

[0151] Among them, the functional proteoglycan is prepared from the following weight ratio of mushroom residue mixture: shiitake mushroom residue 55%, shiitake mushroom residue 20%, and agaric mushroom residue 25%.

[0152] Preparation method:

[0153] (1) Residue pretreatment: Mix shiitake mushroom residue, shiitake mushroom residue, and agaric mushroom residue at a weight ratio of 55:20:25, crush to a particle size of 0.4mm, add 0.6% NaOH solution (solid-liquid ratio 1:6.5), control pH 9.5, pretreat at 65°C for 2.5 hours, stirring speed 160 rpm;

[0154] (2) Ultrasonic-assisted enzymatic hydrolysis: A complex enzyme system (cellulase, hemicellulase, and protease with a mass ratio of 1.8:1.8:1) was added to the pretreated mixed bacteria residue, the enzyme addition amount was 2.2% of the substrate mass, the pH value was adjusted to 6.0, the temperature was controlled at 50°C, and ultrasonic treatment (power 600W, frequency 45kHz) was applied at the same time. The intermittent mode (on for 2.2 minutes, pause for 1.2 minutes) was adopted, and the total ultrasonic treatment time was 42 minutes, and the enzymatic hydrolysis time was 6 hours;

[0155] (3) Multistage membrane separation: The enzyme hydrolysate was centrifuged (5200 rpm, 16 minutes), and the supernatant was sequentially passed through hollow fiber membranes with a molecular weight cutoff of 100 kDa, 50 kDa, and 5 kDa. The operating pressure was 0.4 MPa, the temperature was 30°C, the pH value was 7.0, and the proteoglycan component with a molecular weight of 5-50 kDa was collected;

[0156] (4) Concentration and drying: The collected proteoglycan solution was concentrated to 22% concentration using a rotary evaporator, and then spray-dried (inlet temperature 190°C, outlet temperature 83°C, feed rate 19 L / h) to produce proteoglycan powder;

[0157] (5) Formula mixing: 45 kg of proteoglycan powder, 20 kg of humic acid, 10 kg of potassium dihydrogen phosphate, 15 kg of biochar, and 10 kg of seaweed extract were put into a conical mixer, mixed for 28 minutes at a speed of 23 rpm, and the mixing sequence was the same as in Example 1;

[0158] (6) Granulation and packaging: The mixed material was made into granules by extrusion granulation equipment (pore size 3.5 mm), dried at 64°C for 6.5 hours to a moisture content of 9%, cooled to room temperature, and sieved to collect 2-5 mm granules, and packaged into composite plastic bags, each bag containing 20 kg.

[0159] Product characteristics:

[0160] Appearance: Brown-black granular, no odor

[0161] pH value: 7.8

[0162] Organic matter content: 51.5%

[0163] Total nutrient (N+P2O5+K2O) content: 9.5%

[0164] Moisture content: 9%

[0165] Particle size: 2-5 mm.

[0166] Example 6: Organic fertilizer suitable for alkaline soil

[0167] The organic fertilizer of the present embodiment is composed of the following components in percentage by weight: functional proteoglycan 42%, humic acid 28%, potassium dihydrogen phosphate 14%, biochar 6%, and seaweed extract 10%.

[0168] The functional proteoglycan is prepared from the following mixture of mushroom residues in weight ratio: Lentinula edodes residue 52%, Pleurotus ostreatus residue 23%, and Auricularia auricula residue 25%.

[0169] Preparation method:

[0170] (1) Residue pretreatment: Lentinula edodes residue, Pleurotus ostreatus residue, and Auricularia auricula residue are mixed in a weight ratio of 52:23:25, crushed to a particle size of 0.35 mm, 0.35% NaOH solution (solid-liquid ratio 1:7.5) is added, the pH value is controlled at 8.8, and the pretreatment is carried out at 63℃ for 2.3 hours with a stirring speed of 145 rpm;

[0171] (2) Ultrasonic-assisted enzymatic hydrolysis: a complex enzyme system (cellulase, hemicellulase, and protease in a mass ratio of 2.2:1.8:1.2) is added to the pretreated mixed mushroom residue, the enzyme addition amount is 1.7% of the substrate mass, the pH value is adjusted to 5.5, the temperature is controlled at 46℃, and ultrasonic treatment (power 480W, frequency 38kHz) is applied at the same time in an intermittent manner (turned on for 2.8 minutes and paused for 1.3 minutes), the total ultrasonic treatment time is 38 minutes, and the enzymatic hydrolysis time is 5.8 hours;

[0172] (3) Multistage membrane separation: the enzymatic hydrolysate is centrifuged (5100 rpm, 17 minutes), the supernatant is sequentially passed through hollow fiber membranes with a molecular weight cut-off of 100kDa, 50kDa, and 5kDa, the operating pressure is 0.32MPa, the temperature is 29℃, the pH value is 6.7, and the proteoglycan component with a molecular weight of 5-50kDa is collected;

[0173] (4) Concentration and drying: the collected proteoglycan solution is concentrated to a concentration of 19% using a rotary evaporator, and then spray-dried (inlet temperature 183℃, outlet temperature 82℃, feed rate 21L / h) to obtain proteoglycan powder;

[0174] (5) Formula mixing: 42kg of proteoglycan powder is mixed with 28kg of humic acid, 14kg of potassium dihydrogen phosphate, 6kg of biochar, and 10kg of seaweed extract in a double-cone mixer, the mixing time is 26 minutes, the rotation speed is 21 rpm, and the mixing sequence is the same as in Example 1;

[0175] (6) Granulation and packaging: the mixed material is granulated by extrusion granulation equipment (pore size 2.8mm), dried at 61℃ for 6 hours to a moisture content of 10.5%, cooled to room temperature, sieved, and the 1-4mm granules are collected and packaged into kraft paper bags, each bag containing 15kg.

[0176] Product characteristics:

[0177] Appearance: black granules, no odor

[0178] pH: 5.5

[0179] Organic matter content: 54.2%

[0180] Total nutrient (N+P2O5+K2O) content: 11.2%

[0181] Moisture content: 10.5%

[0182] Particle size: 1-4 mm.

[0183] Comparative Example 1: Lentinula edodes residue organic fertilizer prepared by traditional acid hydrolysis method

[0184] The organic fertilizer of this comparative example is composed of the following components by weight percentage: functional proteoglycan 50%, humic acid 25%, potassium dihydrogen phosphate 10%, biochar 8%, and seaweed extract 7%. The same formula as Example 1, but a different preparation method is used.

[0185] Preparation method:

[0186] (1) Residue treatment: The Lentinula edodes residue is crushed to a particle size of 0.4 mm, 3% hydrochloric acid solution (solid-liquid ratio 1:8) is added, and direct acid hydrolysis is carried out at 95°C for 4 hours;

[0187] (2) Separation and extraction: The acid hydrolysis solution is neutralized (add NaOH solution to pH 7.0), centrifuged, the supernatant is taken, 3 times the volume of 95% ethanol is added for precipitation, and it is placed at 4°C for 12 hours;

[0188] (3) Collect the product: Centrifugal collection of precipitate, 75% ethanol washing 3 times, vacuum drying to get crude proteoglycan product;

[0189] (4) Formula mixing: Mix 50 kg of proteoglycan powder with 25 kg of humic acid, 10 kg of potassium dihydrogen phosphate, 8 kg of biochar, and 7 kg of seaweed extract evenly;

[0190] (5) Granulation and packaging: The same granulation and packaging method as Example 1.

[0191] Compared with Example 1, this comparative example uses the traditional acid hydrolysis method, although it can also extract proteoglycan, but due to high temperature acid treatment, the structure of proteoglycan is damaged, the content of β-1,3 / 1,6-glucan is reduced, and the activity is significantly decreased. The plant immune activation effect of the product is about 40% lower than that of Example 1. At the same time, this method has high energy consumption, produces a large amount of acidic waste liquid, has poor environmental friendliness, and has high extraction cost.

[0192] Comparative Example 2: Organic fertilizer prepared by single strain

[0193] The organic fertilizer of the present comparative example is composed of the following components in weight percentage: functional proteoglycan 50%, humic acid 25%, monopotassium phosphate 10%, biochar 8%, seaweed extract 7%. Same formulation as Example 1, but the functional proteoglycan is prepared only from Lentinula edodes residue (100%).

[0194] Preparation method: The same ultrasonic-assisted enzymatic hydrolysis-multistage membrane separation process as Example 1 is adopted, but the raw material is only Lentinula edodes residue.

[0195] The present comparative example only uses a single strain, and the obtained proteoglycan structure is single, and the function is not comprehensive enough, and the adaptability to different crops and diseases is poor. Experiments have proved that the effect of proteoglycan extracted by a single strain in improving plant disease resistance is 15-20% lower than that of mixed strains. At the same time, the applicability of the product is narrow, and it cannot meet the needs of various crops.

[0196] Comparative Example 3: Organic fertilizer without fractionation

[0197] The organic fertilizer of the present comparative example is composed of the following components in weight percentage: functional proteoglycan 50%, humic acid 25%, monopotassium phosphate 10%, biochar 8%, seaweed extract 7%. Same formulation as Example 1, but the preparation process of functional proteoglycan omits the multistage membrane separation step.

[0198] Preparation method: A similar process to Example 1 is adopted, but after ultrasonic-assisted enzymatic hydrolysis, direct alcohol precipitation is carried out without molecular weight fractionation.

[0199] The present comparative example omits the multistage membrane separation step, and the obtained proteoglycan has a wide molecular weight distribution range (5-300 kDa), among which there are more low-activity and high-molecular-weight components, resulting in low effective ingredient content and about 30% lower plant immune activation effect of the product than Example 1. At the same time, due to the absence of molecular weight fractionation, the quality stability of the product is poor, and there is a large difference between batches.

[0200] Comparative Example 4: Organic fertilizer without biochar formulation

[0201] The organic fertilizer of the present comparative example is composed of the following components in weight percentage: functional proteoglycan 55%, humic acid 30%, monopotassium phosphate 10%, seaweed extract 5%. Compared with Example 1, the biochar component is removed, and the proportion of functional proteoglycan and humic acid is correspondingly increased.

[0202] Preparation method: The same process as Example 1 is adopted, but no biochar is added during the formulation mixing stage.

[0203] The absence of biochar component in the present comparative example leads to a significant reduction in the soil improvement effect of the product, especially in improving soil structure, increasing soil aeration and water retention. The absence of biochar also reduces the product's adsorption capacity for heavy metals and weakens its environmental remediation function. Field trials show that the present comparative example is about 35% less effective than Example 1 in improving soil aggregate structure, and about 25% less effective in water and fertilizer retention.

[0204] Comparative Example 5: Organic fertilizer without seaweed extract formula

[0205] The organic fertilizer of the present comparative example is composed of the following components in weight percentage: functional proteoglycan 54%, humic acid 28%, monopotassium phosphate 10%, and biochar 8%. Compared with Example 1, the seaweed extract component is removed, and the proportion of functional proteoglycan and humic acid is correspondingly increased.

[0206] Preparation method: the same process as Example 1 is adopted, but no seaweed extract is added during the formula mixing stage.

[0207] The absence of seaweed extract component in the present comparative example leads to a lack of bioactive substances such as alginate and mannitol in the product, reducing the product's function of promoting plant growth and stress resistance. The absence of seaweed extract weakens the product's ability to regulate plant hormone balance and reduces its effect on abiotic stress resistance. Field trials show that the present comparative example is about 20% less effective than Example 1 in promoting plant growth, and about 30% less effective in enhancing the drought and cold resistance of crops.

[0208] The effectiveness verification is as follows:

[0209] 1. Comparison of proteoglycan extraction efficiency

[0210] The ultrasonic-assisted enzymatic hydrolysis-multistage membrane separation process of the present application is compared with the traditional acid hydrolysis method (Comparative Example 1), and the results are as follows:

[0211]

[0212] The results show that the method of the present application not only has high extraction efficiency (increased by 82%), but also has high content of active ingredients, low energy consumption (reduced by 39%), and low waste liquid production (reduced by 88%), with obvious technical advantages.

[0213] 2. Plant disease resistance test

[0214] A tomato variety susceptible to disease is selected as the test material, and a potting method is used, with 6 treatments: ① Example 1; ② Example 2; ③ Comparative Example 1; ④ Comparative Example 2; ⑤ Comparative Example 3; ⑥ Control (conventional organic fertilizer). The application amount is 2% of the pot soil. At the 3-leaf stage of the tomato, Botrytis cinerea is inoculated, and the disease index is counted after 7 days, with the following results:

[0215]

[0216] The results show that the product has a significant plant disease resistance effect, and the prevention effects of Example 1 and Example 2 are 78.5% and 81.6% respectively, which are 23.2-38.8 percentage points higher than those of Comparative Examples 1-3. This is mainly due to the use of ultrasonic-assisted enzymatic hydrolysis-multistage membrane separation process in the application, which maintains the integrity of the proteoglycan structure, and the mixed multi-strain provides a variety of active ingredients.

[0217] 3. Soil improvement effect test

[0218] The alkaline soil with pH value of 8.2 is selected for potting test, and the product of Example 6 is applied, the application amount is 3% of the soil, and tomato is planted, and the soil physical and chemical indexes are measured after 60 days, and the results are as follows:

[0219]

[0220] The results show that the product has a significant improvement effect on alkaline soil, can effectively reduce the soil pH value, increase the organic matter content, improve the soil structure, improve the nutrient availability and microbial activity, and create a good soil environment for crop growth.

[0221] The same test is also used to evaluate Comparative Example 4 and Comparative Example 5, and the results show that their effects on improving soil aggregate structure and improving soil microbial activity are significantly lower than those of Example 6.

[0222] 4. Yield and quality effect test

[0223] Tomato yield test is carried out under field conditions, and Comparative Example 1 is compared with Comparative Examples 1, 2 and conventional organic fertilizer, and the application amount is 50 kg / acre, and the results are as follows:

[0224]

[0225] The results show that the product has a significant effect on improving crop yield and quality, and the tomato yield of Example 1 is increased by 17.7%, 11.2% and 29.9% respectively compared with Comparative Examples 1, 2 and conventional organic fertilizer, and the quality index is obviously improved. This is mainly due to the functional proteoglycan in the product, which not only improves the plant disease resistance, but also activates the plant growth-related pathway, promotes the plant growth and development, and improves the yield and quality.

[0226] 5. Application method effect comparison

[0227] The product of Example 1 is selected, and the effects of different application methods on the prevention effect of tomato gray mold and yield are compared, and the results are as follows:

[0228]

[0229] The results show that different application methods have a significant impact on product effect, among which the comprehensive application (basic application + additional application + foliar spraying) has the best effect, the prevention effect reaches 82.1%, and the yield increase rate reaches 30.8%. This shows that the product has adaptability to various application methods, and can be flexibly applied according to different crops and growth stages.

[0230] The functional proteoglycan organic fertilizer prepared from Lentinula edodes residue has simple and feasible preparation process, raw materials are easy to obtain, and the equipment is common, so that large-scale production can be realized. The main raw materials, Lentinula edodes residue, Pleurotus ostreatus residue and Auricularia auricular residue, are all wastes of edible fungus industry, so that the source is wide and the cost is low. The ultrasonic-assisted enzymolysis and multi-stage membrane separation technology used in the preparation process are mature technologies, so that the equipment investment is moderate and the operation is simple. The product has multiple application methods, wide application range and broad market prospect.

[0231] The product can be applied to the cultivation of various crops such as vegetables, fruit trees and economic crops, can improve the disease resistance and yield of crops, improve the soil environment, reduce the use of chemical pesticides, meet the development direction of green agriculture, and has significant economic benefit, social benefit and environmental benefit.

[0232] The present application provides a functional proteoglycan organic fertilizer prepared from Lentinula edodes residue and a preparation method thereof. Through the innovative ultrasonic-assisted enzymolysis and multi-stage membrane separation process, the proteoglycan in the waste residue of edible fungus is efficiently extracted, and then compounded with humic acid, potassium dihydrogen phosphate, biochar and seaweed extract to prepare an organic fertilizer with double functions of promoting growth and activating immunity. The organic fertilizer can not only provide nutrients required for plant growth, but also activate the plant immune system, improve the disease resistance of plants, and at the same time improve the soil physical and chemical properties and soil fertility. The product is suitable for various soil types and crop varieties, has a wide application prospect, and provides a new way for the high-value utilization of waste residue of edible fungus and the development of green agriculture.

Claims

1. A functional proteoglycan organic fertilizer made from shiitake mushroom residue, characterized in that, The organic fertilizer is composed of the following components by weight percentage: Functional proteoglycans 40-60%, 15-30% humic acid Potassium dihydrogen phosphate 8-15%, Biochar 5-10%, 5-10% seaweed extract; The functional proteoglycan is a proteoglycan with a molecular weight of 5-50 kDa, containing a β-1,3 / 1,6-glucan structure; The functional proteoglycan is prepared from a mixture of mushroom residue in the following weight ratios: 50-70% shiitake mushroom residue, 15-25% oyster mushroom residue, and 15-25% wood ear mushroom residue; The method for preparing the functional proteoglycan organic fertilizer from shiitake mushroom residue includes the following steps: (1) Pretreatment of mushroom residue: Mix shiitake mushroom residue, oyster mushroom residue and wood ear mushroom residue, crush to a particle size ≤0.5mm, add 0.3-0.8% NaOH solution, control the pH value to 8.0-9.5, and pretreat at 55-65℃ for 1.5-2.5 hours; (2) Ultrasonic-assisted enzymatic hydrolysis: Add a complex enzyme system to the pretreated mixed bacterial residue. The complex enzyme system includes cellulase, hemicellulase and protease. The amount of enzyme added is 1-3% of the substrate mass. Adjust the pH value to 5.5-6.0, control the temperature at 45-50℃, and apply ultrasonic treatment at a power of 400-600W and a frequency of 35-45kHz. The enzymatic hydrolysis time is 4-6 hours. (3) Multistage membrane separation: After solid-liquid separation of the enzymatic hydrolysate, the supernatant is passed sequentially through membrane separation devices with molecular weight cutoffs of 100kDa, 50kDa, and 5kDa to collect the proteoglycan component of 5-50kDa. (4) Concentration and drying: The collected proteoglycan solution is concentrated to a concentration of 15-25%, and spray drying is carried out with an inlet temperature of 170-190℃ and an outlet temperature of 75-85℃ to produce proteoglycan powder. (5) Formulation mixing: Mix the proteoglycan powder with humic acid, potassium dihydrogen phosphate, biochar and seaweed extract evenly according to the weight percentages stated above; (6) Granulation and packaging: The mixture is made into granules with a particle size of 1-5mm by extrusion granulation equipment, dried to a moisture content of ≤15%, cooled, screened and packaged.

2. The mushroom residue functional proteoglycan organic fertilizer according to claim 1, characterized in that, The physicochemical properties of the organic fertilizer are as follows: pH value 5.0-9.0, Organic matter content ≥45%, Total nutrients Content ≥8%, Moisture content ≤15%, Particle size 1-5mm.

3. The mushroom residue functional proteoglycan organic fertilizer according to claim 1, characterized in that, The humic acid is rich in carboxyl and phenolic hydroxyl groups, exhibiting strong complexing ability; the biochar has a specific surface area ≥300. The porosity is ≥60%; the seaweed extract contains ≥20% alginic acid and ≥5% mannitol.

4. The mushroom residue functional proteoglycan organic fertilizer according to claim 1, characterized in that, In step (1), the pretreatment is carried out in a reaction vessel with a stirring speed of 100-200 rpm and a solid-liquid ratio of 1:5-1:

8.

5. The mushroom residue functional proteoglycan organic fertilizer according to claim 1, characterized in that, In step (2), the ultrasonic treatment is performed intermittently, starting for 2-3 minutes and then pausing for 1-2 minutes, repeating the cycle. The total ultrasonic treatment time is 30-45 minutes.

6. The mushroom residue functional proteoglycan organic fertilizer according to claim 1, characterized in that, In step (3), the membrane separation operation pressure is 0.2-0.4 MPa, the temperature is 25-30℃, and the pH value is controlled at 6.0-7.

0.

7. The mushroom residue functional proteoglycan organic fertilizer according to claim 1, characterized in that, In step (5), a V-type mixer is used for mixing. The mixing time is 15-30 minutes and the speed is 15-25 rpm. First, the proteoglycan powder is mixed with humic acid, then potassium dihydrogen phosphate is added, and finally biochar and seaweed extract are added.

8. The method of applying the functional proteoglycan organic fertilizer from shiitake mushroom residue according to any one of claims 1-7, characterized in that, Includes the following steps: (1) Basic application: 7-15 days before sowing or transplanting crops, spread organic fertilizer evenly on the tillage layer and plow and mix it into the soil. The application rate is 30-50 kg / mu. (2) Additional application: During the critical growth period of crops, combine water and fertilizer application with irrigation system, and dilute organic fertilizer 500-1000 times before use; (3) Foliar spraying: 7-10 days before the peak period of disease, dilute the organic fertilizer 1000-2000 times and spray it evenly on the plant leaves. Repeat every 7-15 days for 2-3 times. It is suitable for various soil types with a pH value of 5.0-9.0, and is applicable to crops including vegetables, fruit trees and cash crops.

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