Cascade coupling HEM functional bacteria bio-organic fertilizer system construction and time sequence combined application method

Abstract

The application discloses a cascade coupling HEM functional bacteria bio-organic fertilizer system construction and time sequence combined application method, and belongs to the technical field of biological fertilizer and soil micro-ecological improvement. In view of the fact that the existing biological fertilizer is simple physical compounding at the factory end, the strain storage is seriously antagonized, the bacterial body preparation is greatly damaged, the fertilizer supply is mismatched with the crop growth period demand, and the bacteria-fertilizer compounding can only realize the reduction of chemical fertilizer, and it is difficult to stably complete the full amount substitution of chemical fertilizer, the application constructs three differentiated functional bacteria modules in time sequence orientation, and matches a substrate type functional water-soluble fertilizer; the bacteria agent modules adopt independent liquid-solid grading fermentation and low-temperature low-damage post-treatment process, are independently packaged when leaving the factory, and are subjected to mild step-by-step coupling compounding before application; time sequence combined application is carried out according to the crop seedling stage-nutrition growth period-reproductive growth period, so that the microbial community completes cascade coupling metabolism in situ in the rhizosphere soil, the metabolic products of the previous microbial community are used as the growth substrate of the subsequent microbial community, and a metabolic relay chain is formed; the matched water-soluble fertilizer is mainly used as the carbon and nitrogen substrate for microbial community proliferation and rhizosphere signal material, relies on soil organic matter mineralization and microbial community metabolism to supply nutrients to crops in the whole growth period, and realizes full amount substitution of chemical nitrogen, phosphorus and potassium fertilizer under field conditions. The application can improve soil organic matter, reduce the occurrence rate of soil-borne diseases, and keep the crop yield flat with the conventional chemical fertilizer treatment; the process can be industrialized and enlarged, and is suitable for large-scale planting of various crops such as field grain, facility fruits and vegetables and economic Chinese medicinal materials.

Description

Technical Field

[0001] This invention belongs to the fields of bio-organic fertilizer preparation, soil micro-ecological improvement, and green agricultural fertilization technology. Specifically, it involves a fully chemical fertilizer replacement cultivation technology system that integrates modular construction of HEM multifunctional microbial communities, graded low-temperature coupling preparation, and sequential application throughout the entire crop growth period. It is particularly suitable for ecological restoration and quality improvement and yield enhancement of farmland soils that are degraded by continuous cropping, have secondary salinization, or are compacted and acidified. It can be widely applied to green planting of greenhouse fruits and vegetables, field grains, and cash crops with zero chemical fertilizer input, and belongs to the field of agricultural microbial fertilizer and ecological fertilization technology innovation. Background Technology

[0002] Currently, my country's arable land generally suffers from problems such as long-term continuous cropping, excessive application of nitrogen, phosphorus, and potassium fertilizers, insufficient input of organic fertilizers, and shallow topsoil. This leads to a series of degradation phenomena in farmland soils, including secondary salinization, pH imbalance, destruction of soil aggregate structure, lack of organic matter, and a sharp decline in soil microbial diversity. The imbalance of soil microecology further triggers production problems such as outbreaks of soil-borne diseases, high incidence of root-knot nematodes, poor crop root vitality, stunted and weak seedlings, unstable yields, and declining quality, seriously hindering the high-quality development of facility agriculture and large-scale field agriculture.

[0003] Existing commercial bio-organic fertilizers, compound microbial agents, and related systems suffer from numerous structural and technical defects in practical applications, which can be summarized into the following core technical bottlenecks: First, traditional microbial fertilizers often employ a multi-strain physical premixing process, mixing functional strains with different functions, metabolic habits, and growth conditions in a single batch at the factory. Because different strains exhibit significant differences in their proliferation cycles, metabolic substrate requirements, and salt and temperature tolerance, premixed storage is highly susceptible to bacterial antagonism, metabolic competition, and the death of dominant strains. This results in rapid decay of live bacteria in the finished product, short shelf life, and low field colonization rates. Consequently, the actual fertilizer application effect is far lower than the theoretical laboratory effect, a common technical pain point in the industry.

[0004] Secondly, existing bio-fertilizer preparation processes generally suffer from defects such as high-temperature drying, high-intensity mechanical mixing, and simultaneous compounding of high-salt nutrients. Traditional drying temperatures are generally above 60℃, which can damage spore cells, rupture cell membranes, and inactivate functional metabolic proteins, significantly reducing the survival rate of effective live bacteria. At the same time, the one-time mixing of high-salt trace elements, high-concentration organic matter, and bacterial powder creates a high-osmotic-pressure environment that further kills functional bacteria, resulting in a low rate of effective live bacteria count in the finished product and poor field resistance and survival ability.

[0005] Third, existing fertilization systems all adopt a one-time basal application and static fertilization model, which cannot match the dynamic nutritional needs of crops throughout their entire growth period and the succession patterns of the rhizosphere microecology. From the seedling stage, vegetative growth stage to the reproductive growth stage, there are significant differences in the root distribution range, nutrient absorption patterns, and the degree of stress in the rhizosphere microenvironment. Single fertilizers and single fertilization cannot achieve the segmented functional matching of "root protection and antibacterial activity in the seedling stage, nutrient activation in the growth stage, and stress resistance and stable yield in the fruit expansion stage". This easily leads to problems such as redundant competition of microbial communities in the early stage, absence of functional microbial communities in the later stage, and nutrient supply gaps.

[0006] Fourth, existing combinations of microbial fertilizers and water-soluble fertilizers are mostly simple superpositions without scientific sequential matching and metabolic coupling design. Conventional solutions simply package solid microbial fertilizers and water-soluble fertilizers separately and apply them randomly, failing to construct a closed-loop metabolic logic of "microbial colonization - substrate supply - nutrient activation - crop absorption." Water-soluble fertilizers only serve as a nutrient supplement for crops and cannot act as a dedicated carbon and nitrogen substrate for the proliferation and metabolism of functional microorganisms. This makes it difficult to achieve continuous optimization of the soil microecology and long-term nutrient activation. Therefore, existing technologies can only reduce the amount of chemical fertilizers used and cannot achieve large-scale, stable, and complete replacement of chemical fertilizers.

[0007] Fifth, existing technologies lack a systematic microbial fertilizer construction system that is functionally modular, time-adaptable, and metabolically coupled. Most microbial fertilizers on the market have only one function, possessing only simple soil improvement or growth promotion functions. They cannot simultaneously achieve multiple functions such as soil remediation, rhizosphere protection, disease control, nutrient activation, and stress resistance and yield increase. They cannot form a systematic remediation solution for degraded soils such as those affected by continuous cropping, saline-alkali soils, and compacted soils, and their technological adaptability and scenario compatibility are poor.

[0008] In summary, existing bio-fertilizers generally suffer from technical defects such as imperfect formulation systems, damage to microorganisms during preparation processes, severe antagonism among microbial communities, static and fixed application patterns, poor coupling between microorganisms and fertilizers, and inability to fully replace chemical fertilizers. The industry urgently needs a new technical system that can achieve modular independent preparation of microbial communities, low-damage coupled compounding, sequential application during the growth period, and in-situ cascade metabolism in the soil, breaking through the existing technical bottlenecks from four dimensions: fertilizer structure, preparation process, application logic, and ecological metabolism.

[0009] Based on this, the present invention proposes a cascaded coupling HEM functional microbial bio-organic fertilizer system construction and sequential application method. Through the construction of differentiated functional microbial community modules, low-temperature stepwise coupling preparation, and sequential cascade application throughout the entire growth period, the system achieves dynamic restoration of rhizosphere microecology, continuous activation of soil nutrients, and full-process crop nutrient self-sufficiency, truly realizing the ecological restoration of degraded soils due to continuous cropping and green cultivation with full replacement of chemical fertilizers. Summary of the Invention

[0010] The following five major technological bottlenecks exist in the current bio-organic fertilizer field: ① Premixing multiple microbial strains in factories leads to antagonistic competition among microbial communities, resulting in rapid decay of live bacteria during storage and low rhizosphere colonization efficiency in the field; ② Traditional high-temperature drying and high-intensity mixing processes cause significant damage to functional microorganisms, resulting in low retention rates of live bacteria in the finished product; ③ The fertilizer application mode is a one-time basal application, which cannot match the succession patterns of rhizosphere microecology at different growth stages of crops, leading to excessive competition among microbial communities in the early stages and a lack of functional bacteria in the later stages; ④ The microbial-fertilizer combination is merely a simple physical combination, lacking metabolic coupling design. Water-soluble fertilizer is directly used as crop nutrients and cannot serve as a substrate for microbial community proliferation, only achieving fertilizer reduction, which is difficult to stably achieve full replacement of chemical fertilizers under field conditions; ⑤ Existing products have fragmented functions and lack modular systems, resulting in limited ability to remediate acidified, salinized, and continuously cropped degraded soils.

[0011] The technical problem to be solved by this invention is to overcome the inherent defects of existing bio-fertilizer formulations, processes, and application systems, and to provide a method for constructing and sequentially applying a cascade-coupled functional microbial bio-organic fertilizer system; to solve the technical problems of multi-strain coexistence and antagonism, microbial cell damage during preparation, mismatch between fertilizer supply and crop growth period, metabolic disconnect between microbial fertilizer and water-soluble fertilizer, and difficulty in fully replacing chemical fertilizers; and to achieve the micro-ecological restoration of degraded farmland under field and facility cultivation conditions, and to stably achieve a zero-input planting mode of chemical nitrogen, phosphorus, and potassium fertilizers.

[0012] The first objective of this invention is to construct a functionally partitioned modular microbial community system, abandoning the traditional approach of mixing multiple microorganisms in one pot, and dividing functional modules according to the logic of rhizosphere ecological succession, thereby avoiding the risk of long-term antagonism between strains from the source of the formula.

[0013] Secondly, it provides a low-damage, graded coupling preparation process, in which each functional bacterial module is fermented and propagated independently. Low-temperature post-treatment and mild stepwise compounding processes are used to maximize the preservation of the physiological activity of each module strain and improve the shelf-life viable bacteria retention rate of the product. The third aspect is to change the traditional idea of ​​completing microbial community coupling in factories and to innovatively adopt the soil in-situ cascade coupling metabolic mechanism. Instead of forcibly completing microbial community synergy at the production end, it is transferred to the rhizosphere soil environment and applied sequentially according to the crop growth cycle to achieve the successive colonization of microbial communities, with metabolic products serving as substrates for each other, and the functions being carried out in a step-by-step relay. Fourthly, it involves reconstructing the synergistic logic of bacteria and water-soluble fertilizer, positioning the matching amino acid-humic acid water-soluble fertilizer as a substrate for bacterial metabolism and a substance for inducing rhizosphere signals, rather than directly supplying crops with large amounts of inorganic nutrients. This establishes a complete closed loop of "modular bacterial agent colonization - substrate supply - organic matter mineralization - nutrient release - crop absorption," achieving full replacement of chemical fertilizers under field conditions. Fifthly, it provides a complete set of technical solutions that are suitable for grains, fruits, vegetables, and cash crops, taking into account multiple effects such as soil improvement, suppression of soil-borne diseases, stress resistance and quality improvement, and stable and increased yield. The process is easy to scale up industrially and is suitable for large-scale agricultural production scenarios.

[0014] To achieve the above-mentioned objectives, the present invention adopts the following technical solution:

[0015] A method for constructing and sequentially applying a cascaded coupled HEM functional bacteria bio-organic fertilizer system includes system construction steps, a cascaded coupling preparation step, and a sequential application step.

[0016] The system construction steps are as follows: a modular complete fertilizer system is constructed, which includes three functionally differentiated independent microbial agent modules and supporting functional water-soluble fertilizer components; the three microbial agent modules are divided according to the temporal logic of crop rhizosphere microecology establishment - organic matter nutrient activation - stress resistance regulation, namely the rhizosphere activation bacteria module, the organic matter mineralization bacteria module, and the stress resistance regulation bacteria module.

[0017] Rhizosphere activating bacteria module: responsible for modifying the microenvironment around the roots in the early stages of transplanting / sowing, prioritizing the occupation of the rhizosphere ecological niche, constructing colonization sites, and providing a survival basis for the subsequent entry of other microbial communities; Organic matter mineralizing bacteria module: responsible for the mineralization and decomposition of inherent organic matter and organic carriers in the soil, continuously releasing slow-release nitrogen, phosphorus, potassium and trace nutrients to supply crops with nutrition; Stress resistance regulating bacteria module: planted on the root surface during the reproductive growth stage, it induces systemic resistance in crops, inhibits soil-borne harmful microorganisms, alleviates stress from continuous cropping, salinity, and drought, and ensures stable yield and improved quality during the reproductive growth stage.

[0018] The supporting functional water-soluble fertilizer components are: amino acid-humic acid type substrate water-soluble fertilizer, which does not contain inorganic nitrogen, phosphorus and potassium main fertilizers. It provides carbon and nitrogen metabolism substrates for the proliferation of functional bacteria in each module, and at the same time secretes rhizosphere signaling substances to promote the adsorption and colonization of functional bacteria. It does not independently undertake the supply of main nutrients to crops.

[0019] The graded coupling preparation steps are as follows: each of the three microbial agent modules undergoes independent liquid seed fermentation, solid carrier propagation, and low-temperature air drying to obtain independently stored microbial agent units; only before application or at the application site is the microbial agent compounded with the well-rotted organic matter carrier at room temperature and low speed in a stepwise coupling process; high-temperature drying and high-intensity shear mixing are avoided throughout the process to maximize the protection of microbial activity; each module is packaged independently at the factory without deep premixing, thus inhibiting antagonistic competition between strains during the storage stage from the source.

[0020] Furthermore, each microbial agent module employs an independent fermentation process: (1) Primary liquid seed fermentation: Each group of functional strains is inoculated into the corresponding liquid culture medium, the temperature is 28-32℃, the dissolved oxygen is controlled at 25-40%, and the fermentation time is 24-36h to obtain a highly active seed liquid; each strain is fermented separately and is not mixed with other strains in the same tank. (2) Secondary solid carrier propagation: Spray the corresponding seed liquid onto the sterilized organic carrier material with a moisture content of 42-48%. Perform aerobic fermentation at 26-30℃ for 4-6 days in a closed environment. Turn the pile regularly to replenish oxygen and complete the solid propagation to obtain a single-module solid substrate. (3) Low-temperature passivation post-treatment: After fermentation, the material is air-dried at a low temperature of 40-45℃. The material temperature is strictly controlled not to exceed 50℃ to obtain independent microbial agent module units, which are then individually sealed and stored in separate bags.

[0021] Furthermore, the step-by-step coupling compounding process is as follows: compounding is carried out before field application, with an ambient temperature of 20-35℃, a stirring speed of 20-35r / min, and a mixing time of 8-12min. Low-speed and light stirring is used to achieve only uniform mixing of materials and avoid mechanical shearing damage to the spore cells. No granulation high-temperature process is carried out to preserve the original metabolic characteristics of each microbial community module.

[0022] Furthermore, the viable bacteria indicators for each module are as follows: The rhizosphere activated bacteria module includes Bacillus subtilis and Bacillus amyloliquefaciens, with an effective viable bacteria count ≥ 2.2 × 10⁻⁶. 8 CFU / g; The organic matter mineralizing bacteria module contains Bacillus megaterium and Bacillus mucilaginosus, with an effective viable count ≥ 2.1 × 10⁻⁶. 8 cfu / g; the stress resistance regulating bacteria module includes Pseudomonas and Trichoderma, with an effective viable count ≥2.0×10⁻⁶. 9 cfu / g.

[0023] The accompanying water-soluble fertilizer contains ≥120g / L total amino acids and ≥50g / L humic acid, with no external addition of large amounts of inorganic nitrogen, phosphorus, and potassium.

[0024] The sequential application steps are as follows: based on the crop's growth rhythm from seedling stage to vegetative growth stage to reproductive growth stage, corresponding microbial agent modules are added in stages, simultaneously combined with drip irrigation / fertigation of matching functional water-soluble fertilizers; after the microbial community enters the soil, it completes sequential colonization and cascade coupled metabolism in the rhizosphere in situ environment; the metabolic products of the microbial community in the previous stage serve as the substrate for the growth of the microbial community in the next stage, forming a metabolic relay chain; no chemical nitrogen, phosphorus and potassium fertilizers are applied throughout the process, relying on soil organic matter mineralization + microbial community coupled metabolism to supply nutrients for the crop throughout its entire growth period.

[0025] Specific stages of sequential implementation: a) Early stage of sowing / transplanting: Apply rhizosphere activated bacteria module, in combination with matching functional water-soluble fertilizer, to prioritize the occupation of rhizosphere sites, modify the rhizosphere microenvironment, and lay a good foundation for subsequent colonization of microorganisms. b) Vegetative growth stage: Apply organic matter mineralization bacteria module, combined with matching functional water-soluble fertilizer, to enhance the mineralization and decomposition of soil organic matter, continuously release slow-release nutrients, and meet the nutritional growth needs of crops. c) Reproductive growth stage: Apply stress resistance regulating bacteria modules, along with matching functional water-soluble fertilizers, to regulate the rhizosphere microenvironment for stress resistance, inhibit soil-borne diseases, and ensure development during the flowering, fruit setting, and grain filling stages.

[0026] Field application parameters: 40-60 kg / mu for rhizosphere activating bacteria module, 30-50 kg / mu for organic matter mineralizing bacteria module, and 25-40 kg / mu for stress resistance regulating bacteria module; supplement with 2-5 kg / mu of functional water-soluble fertilizer each time, applied by drip irrigation or fertigation, in 3-5 applications throughout the entire growth period.

[0027] This invention also protects the cascade-coupled HEM functional bacteria bio-organic fertilizer complete fertilizer product prepared by the aforementioned method.

[0028] Formula system innovation: Time-oriented modular microbial community segmentation

[0029] Breaking away from the traditional approach of classifying strains based on simple functions like growth promotion and phosphorus and potassium solubilization, this product is divided into three major functional modules according to the time sequence of rhizosphere ecological succession. These modules have a succession relationship in terms of ecological niches, rather than simply having parallel functions. The product is individually packaged at the factory to avoid storage antagonism and solve the long-standing problem of shelf-life activity decline in multi-strain compound microbial fertilizers.

[0030] Innovative preparation process: modular independent fermentation + low-temperature, low-damage stepwise coupling process The conventional industry process of mixed fermentation and high-temperature granulation and drying was abandoned; each strain module was fermented and propagated separately, and air-dried at low temperature, with only slight compounding before application; process comparison proved that compared with the traditional high-temperature granulation process, the initial viable count of the finished product and the viable retention rate after 3 months of storage were significantly improved, and the process improvement brought clear and quantifiable technical effects.

[0031] Innovative Mechanism of Action: In-situ Cascade Coupling of Metabolism in Soil Existing technologies all attempt to achieve synergistic effects by mixing multiple microorganisms within a factory. This invention innovatively transfers the microbial coupling process to the rhizosphere soil of crops, achieving sequential colonization of the microbial community through sequential application. The metabolic products of preceding applications serve as substrates for subsequent applications, forming a sequential metabolic relay chain that matches the dynamic changes in the crop's growth cycle, thus solving the problem of mismatch between fertilizer supply and crop demand. This mechanism is the most core distinguishing technical feature of this invention compared to existing microbial fertilizer combinations.

[0032] Synergistic Innovation of Microbial Fertilizer and Water-Soluble Fertilizer: Substrate-Based Water-Soluble Fertilizer Design This approach changes the traditional idea of ​​"compound fertilizer directly supplying nutrients to crops." Instead, the water-soluble fertilizer used in the fertilizer is not the main nutrient source. Its core function is to serve as a substrate for the proliferation of carbon and nitrogen in functional bacteria and a rhizosphere signal inducer, driving the proliferation and metabolism of modular microbial communities within the soil. It also mineralizes the background organic matter in the soil to release nutrients, truly achieving full replacement of chemical fertilizers under field conditions, rather than simply reducing the amount of chemical fertilizers.

[0033] This invention is not a simple combination of several fertilizers, but a complete closed-loop system formed from formula construction, fermentation preparation, compounding process, in-situ metabolic mechanism, and time-sequential application method; the various technical features are interconnected and mutually supportive, and are not a simple superposition of existing technical features. Beneficial effects

[0034] Each functional microbial module of this invention is fermented and packaged independently, avoiding antagonistic competition between different strains during storage from the source; the low-temperature, low-damage preparation process significantly increases the initial viable count and shelf-life viable retention rate of the finished product; compared with the traditional high-temperature granulation process, the viable retention rate after 3 months of storage is increased from 31.7% to 82.4%, and the product storage stability is significantly improved.

[0035] The soil-based in-situ cascaded coupled metabolism model is adopted, with the microbial community colonizing sequentially according to the crop's growth stage, and the metabolism relaying. This matches the rhizosphere environment and nutrient requirements of different stages of crop growth, such as seedling stage, vegetative growth, and reproductive growth, avoiding the problems of early niche competition and later functional loss caused by applying all microbial communities at once.

[0036] The synergistic relationship between microbial agents and water-soluble fertilizers was reconstructed, and the substrate-type water-soluble fertilizers were used to drive the mineralization and release of nutrients from the soil's background organic matter. Under field and facility cultivation conditions, it can stably achieve full replacement of chemical nitrogen, phosphorus, and potassium fertilizers. Experiments showed that the yield of corn under the condition of no chemical fertilizer application can reach the same level as that of conventional fertilizer treatment, and the yield of greenhouse tomatoes is not lower than that of the fertilizer control.

[0037] Continuously improve degraded arable land, increase soil organic matter, reduce soil bulk density, inhibit the occurrence of soil-borne diseases, alleviate the harm of continuous cropping and secondary salinization, and take into account multiple benefits such as soil improvement, growth promotion, stress resistance and yield increase.

[0038] The entire preparation process is mild and uses a wide range of raw materials, which can be directly connected to existing organic fertilizer factory equipment to complete industrial production. The application scheme is compatible with mainstream agricultural operations such as broadcasting, fertigation, and drip irrigation, and is suitable for large-scale promotion and application of various crops such as grains, fruits and vegetables, and Chinese medicinal herbs. Attached Figure Description

[0039] Figure 1 This is a flowchart illustrating the overall system construction, hierarchical preparation, and sequential application process of the present invention. Figure 2 This is a block diagram of the in-situ cascaded coupling metabolic logic in soil according to the present invention; Figure 1Process Description: Modular system construction → Independent fermentation and propagation of each module → Low-temperature air drying and independent packaging → Mild stepwise compounding before application → Sequential application throughout the crop's entire growth period → In-situ cascaded coupling metabolism in the rhizosphere → Soil improvement + complete replacement of chemical fertilizers + improved crop quality and yield. The system modules are, in order, rhizosphere activation module A, organic matter mineralization module B, stress regulation module C, and Helm water-soluble fertilizer module D.

[0041] Figure 2 Logical explanation: Seedling stage ecological niche construction → nutrient stage root ecological niche occupation → reproductive stage stress resistance and stable yield, the microbial community metabolism relay, substrate complementarity, and functional progression at each stage form a closed-loop ecological fertilization system. Detailed Implementation

[0042] All experimental data testing standards in this invention strictly follow the standards of "GB / T 20287-2006 Agricultural Microbial Agents", "NY / T 525-2021 Organic Fertilizers", and conventional soil agrochemical testing methods. All field trials uniformly adopt a plot randomized block design with three biological replicates, and set up multiple comparison groups including blank control, conventional fertilizer control, and commercially available microbial fertilizer control.

[0043] The unified preparation standard of this invention: Liquid fermentation temperature: 28-32℃, fermentation time: 24-36h; solid propagation temperature: 26-30℃, moisture content: 42-48%, fermentation time: 4-6d; low-temperature air drying temperature: 40-45℃, maximum not exceeding 50℃; compounding temperature: 20-35℃, rotation speed: 20-35r / min, duration: 8-12min.

[0044] The standard specifications for water-soluble fertilizers are: total amino acids ≥120g / L, humic acid ≥50g / L, and no inorganic nitrogen, phosphorus, or potassium single-element fertilizers.

[0045] Basic conditions of the experimental plot: a typical continuous cornfield in North China, planted for more than 10 years, with soil salinity of 1.86 g / kg, pH 8.2, and organic matter of 14.7 g / kg. Soil-borne diseases are prevalent year after year, and the land is a typical degraded arable land.

[0046] Viable microbial index of the microbial community module: Microbial substrate (rhizosphere activation module) 2.3×10 8 CFU / g; Bacterial-based lyolysis (organic matter mineralization module) 2.1 × 10⁻⁶ 8 CFU / g; Bacterial-based wireless (stress regulation module) 2.6 × 10⁻⁶ 9 cfu / g.

[0047] Application plan (dosage per acre): 1. Sowing and land preparation period: 50 kg of fungal-based fertile soil is evenly spread and tilled to a depth of 22 cm, and 3 kg of Hem water-soluble fertilizer is applied by irrigation. 2. During the jointing and vegetative growth stage: 40 kg of bacterial substrate solution and 4 kg of Hem water-soluble fertilizer for drip irrigation; 3. From the large trumpet mouth to the male reproductive growth stage: 30kg of microbial substrate and 3kg of Hem water-soluble fertilizer for drip irrigation; No chemical nitrogen, phosphorus, or potassium fertilizers are used throughout the entire process.

[0048] Harvest test results: At harvest, soil organic matter increased to 18.6 g / kg, soil bulk density decreased from 1.38 g / cm³ to 1.21 g / cm³, and soil salinity decreased to 1.23 g / kg; the incidence of soil-borne diseases was only 7.2%; the corn yield was 642.7 kg / mu; the yield of commercially available ordinary microbial fertilizer without chemical fertilizer treatment was 481.3 kg / mu, with a disease incidence of 16.8%; the yield of farmers treated with conventional chemical fertilizer was 635.2 kg / mu, and after harvest, the soil organic matter was 14.7 g / kg, the soil bulk density was 1.38 g / cm³, the salinity was 1.72 g / kg, and the disease incidence was 14.5%. As can be seen from the results in Table 1, Example 1 of this invention, without the application of chemical fertilizers throughout the entire process, resulted in a slightly higher corn yield than the conventional chemical fertilizer treatment; soil organic matter was significantly increased, soil bulk density and salinity decreased, and soil-borne diseases were significantly suppressed; under the same organic matter input conditions, the yield of commercially available microbial fertilizer without chemical fertilizers decreased significantly, and it could not achieve a complete replacement of chemical fertilizers.

[0049] Table 1 Comparison of effects among different groups in maize field experiment

[0050] Example 2: Experiment on the cultivation of tomatoes in greenhouses with continuous cropping, completely replacing chemical fertilizers. Experimental conditions: Tomatoes were continuously grown in greenhouses for 8 years, with high incidence of root-knot nematodes and wilt disease, and secondary soil salinization.

[0051] Application plan per mu: Before transplanting, apply 45 kg of fungal-based fertile soil as a base fertilizer; during the seedling establishment and vigorous growth period, apply 35 kg of fungal-based soluble fertilizer + 3 kg of Hem water-soluble fertilizer; during the fruit setting and expansion period, apply 28 kg of fungal-based soluble fertilizer and supplement with Hem water-soluble fertilizer twice, totaling 7.5 kg; zero chemical fertilizer input throughout the entire process.

[0052] Harvest test results: Tomato yield was 5218 kg / mu, higher than the local conventional fertilizer control group's yield of 5106 kg / mu; root-knot nematode incidence decreased from 29.4% to 11.2%; soil organic matter increased by 2.1 g / kg compared to before the experiment; the soluble solids content of the fruit in the invention group was 5.7%; the commercially available microbial fertilizer-free group had an increase of 0.7 g / kg in organic matter, a root-knot nematode incidence of 24.6%, a yield of 4126 kg / mu, and soluble solids content of 4.8%; the conventional fertilizer control group had an increase of 0.3 g / kg in organic matter, a root-knot nematode incidence of 29.4%, a yield of 5106 kg / mu, and soluble solids content of 4.9%. As shown in Table 2, under the condition of completely eliminating chemical fertilizers, the invention achieved a higher tomato yield than the fertilizer control group, while simultaneously restoring the soil, significantly reducing nematode disease, and improving the fruit quality index of soluble solids content, achieving a synergistic effect of stable yield and improved quality.

[0053] Table 2 Comparison of effects among different groups in the greenhouse tomato experiment

[0054] Example 3 Gradient Dosage Adaptation Verification Test Slightly degraded farmland was selected, and three application dose gradients (low, medium, and high) were set to verify the applicability of the parameter range within the scope of protection of the claims.

[0055] Low dose: 40kg of bacterial-based fertile soil + 30kg of bacterial-based solvent + 25kg of bacterial-based wireless; Medium dose: 50kg of bacterial-based fertile soil + 40kg of bacterial-based solvent + 30kg of bacterial-based wireless agent; High dosage: 60kg of bacterial-based fertile soil + 50kg of bacterial-based solvent + 40kg of bacterial-based wireless agent.

[0056] Test results: In the low-dose group, soil organic matter was 16.2 g / kg, the incidence of soil-borne diseases was 11.5%, and the corn yield was 563.4 kg / mu; in the medium-dose group, soil organic matter was 18.1 g / kg, the incidence of soil-borne diseases was 7.6%, and the corn yield was 631.8 kg / mu; in the high-dose group, soil organic matter was 18.9 g / kg, the incidence of soil-borne diseases was 6.8%, and the corn yield was 640.2 kg / mu. The experiment shows that the low dose is suitable for mildly degraded soils; the medium and high doses are more effective in improving moderately to severely degraded soils; proving that the dosage ranges of 40-60 kg, 30-50 kg, and 25-40 kg as claimed in this invention are reasonable and effective.

[0057] Table 3 Soil and yield results under different application gradients

[0058] Comparative Example 1: Preparation Process Comparison and Verification Experiment This comparative example is used to demonstrate the technical advantages of the independent fermentation and low-temperature air-drying process of the present invention compared with the traditional high-temperature granulation process.

[0059] Experimental group: The process of this invention involves independent fermentation of each module, low-temperature air drying at 43℃, and independent packaging; Control group: Traditional process, all strains are mixed and then dried and granulated at 65℃.

[0060] Test results: The initial total viable count of the finished product from the low-temperature independent fermentation process of this invention was 2.0 × 10⁻⁶. 9 CFU / g, viable count after 3 months of storage: 1.65 × 10⁻⁶ 9 CFU / g, 3-month viable cell retention rate 82.4%; initial total effective viable cell count of the finished product from traditional high-temperature mixed-culturing granulation process was 4.3 × 10⁻⁶. 8 CFU / g, viable count after 3 months of storage: 1.36 × 10⁻⁶ 8 The cfu / g and 3-month viable cell retention rate were 31.7%. Table 4 data confirms that traditional high-temperature granulation processes cause significant cell death and severe storage degradation; the process of this invention can significantly retain viable cells and improve shelf-life stability, which is key inventive evidence at the process level.

[0061] Table 4 Comparison of live bacteria preservation effects of different preparation processes

[0062] Combining the experimental data from Tables 1-4, it is clear that this invention addresses the industry pain point that traditional bio-fertilizers cannot stably and completely replace chemical fertilizers through five core technical features: modular independent preparation, low-temperature and low-damage process, in-situ cascaded coupled metabolism in soil, sequential application, and substrate-based water-fertilizer coupling. Under conditions without the application of chemical nitrogen, phosphorus, and potassium fertilizers, it achieves stable crop yields, soil remediation, disease reduction, and improved agricultural product quality. Comparative examples demonstrate that without the core technical features of this invention, the technical effects of this invention cannot be achieved, proving that the entire technical solution is not a simple superposition of existing technologies and possesses sufficient novelty and inventiveness. The technical solution of this invention has already been commercialized and field-verified in the Hemong series of products (Bacterial-Based Fertile Soil, Bacterial-Based Soluble Soil, Bacterial-Based Wireless Soil, and Hemong Water-Soluble Fertilizer) of Shuike Environmental Technology (Zhengzhou) Co., Ltd.

[0063] It will be apparent to those skilled in the art that the present invention is not limited to the details of the exemplary embodiments described above, and that the invention can be implemented in other specific forms without departing from its spirit or essential characteristics. Therefore, the embodiments should be considered in all respects as exemplary and non-limiting, and the scope of the invention is defined by the appended claims rather than the foregoing description. Thus, all variations falling within the meaning and scope of equivalents of the claims are intended to be included within the present invention. No reference numerals in the claims should be construed as limiting the scope of the claims.

[0064] Furthermore, it should be understood that although this specification describes embodiments, not every embodiment contains only one independent technical solution. This narrative style is merely for clarity. Those skilled in the art should consider the specification as a whole, and the technical solutions in each embodiment can also be appropriately combined to form other embodiments that can be understood by those skilled in the art.

Claims

1. A method for constructing and sequentially applying a cascaded coupled HEM functional bacteria bio-organic fertilizer system, characterized in that, This includes system construction steps, hierarchical coupling preparation steps, and time-series combined application steps; The system construction includes: setting up at least 3 functionally differentiated microbial agent modules, and supporting functional water-soluble fertilizer components; the microbial agent modules are a rhizosphere activating bacteria module, an organic matter mineralizing bacteria module, and a stress resistance regulating bacteria module; The graded coupling preparation step includes independently fermenting each microbial agent module to obtain a solid microbial agent unit, and then coupling and compounding it with the carrier organic matter in a stepwise manner, without high-temperature mixing; The sequential application method involves applying various microbial agent modules and supporting functional water-soluble fertilizer components in sequence or in segments according to different growth stages of the crop. It relies on the in-situ coupling metabolism of microbial communities to complete the nutrient supply for the entire growth period of the crop and achieve full replacement of chemical fertilizers.

2. The method according to claim 1, characterized in that, The rhizosphere activated bacteria module contains Bacillus subtilis and Bacillus amyloliquefaciens, with an effective viable count ≥ 2.2 × 10⁻⁶. 8 CFU / g; The organic matter mineralizing bacteria module contains Bacillus megaterium and Bacillus mucilaginosus, with an effective viable count ≥ 2.1 × 10⁻⁶. 8 cfu / g; the stress resistance regulating bacteria module includes Pseudomonas and Trichoderma, with an effective viable count ≥2.0×10⁻⁶. 9 cfu / g.

3. The method according to claim 1, characterized in that, The independent fermentation processes for each microbial agent module are as follows: (1) Primary liquid seed fermentation: The corresponding strains are inoculated into liquid culture medium, the temperature is 28-32℃, the dissolved oxygen is 25-40%, and the fermentation time is 24-36h to obtain seed liquid; each strain is fermented separately and not mixed in the same tank. (2) Secondary solid propagation: Spray the seed liquid onto a sterilized organic carrier, control the moisture content to 42-48%, and ferment in a closed aerobic environment at 26-30℃ for 4-6 days, turning the pile to supply oxygen during the process, to obtain a single-module solid microbial agent; (3) Low-temperature passivation post-treatment: After fermentation, air dry at 40-45℃, the material temperature shall not exceed 50℃, and each microbial agent module unit shall be stored independently and packaged separately.

4. The method according to claim 1, characterized in that, Stepwise coupling and compounding: Each microbial agent module is mixed with the decomposed organic matter carrier at a low speed of 20-35℃, with a stirring speed of 20-35r / min and a mixing time of 8-12min, to avoid mechanical damage to the microbial cells; each module is not pre-mixed at the factory to preserve the independent activity basis of the module's microbial community.

5. The method according to claim 1, characterized in that, The supporting functional water-soluble fertilizer component is an amino acid-humic acid type water-soluble fertilizer with a total amino acid content ≥120g / L and a humic acid content ≥50g / L. It does not contain inorganic nitrogen, phosphorus, and potassium fertilizers. As a substrate for bacterial metabolism and a signaling substance, it works synergistically with the bacterial agent module to supply nutrients.

6. The method according to claim 1, characterized in that, The timing-based simultaneous application includes: a) Pre-sowing / transplanting stage: Apply rhizosphere activating bacteria module, along with some matching functional water-soluble fertilizer, to activate the rhizosphere soil and establish initial colonization sites; b) Vegetative growth period: Apply organic matter mineralizing bacteria module, combined with matching functional water-soluble fertilizer, to mineralize soil organic matter and release slow-release nutrients; c) Reproductive growth period: Apply stress resistance regulating bacteria module, combined with matching functional water-soluble fertilizer, to regulate the rhizosphere stress resistance microenvironment and ensure reproductive growth; No chemical nitrogen, phosphorus, or potassium fertilizers are applied throughout the entire growth process. Nutrient supply is achieved through microbial coupled metabolism and water-soluble fertilizer substrate.

7. The method according to claim 6, characterized in that, Application parameters per mu in the field: 40-60 kg of rhizosphere activating bacteria module, 30-50 kg of organic matter mineralizing bacteria module, and 25-40 kg of stress resistance regulating bacteria module; apply 2-5 kg / mu of matching functional water-soluble fertilizer each time, with drip irrigation or fertigation, in 3-5 applications according to the growth stage.

8. The cascaded coupled functional microbial bio-organic fertilizer product obtained by the method according to any one of claims 1-7.

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