A smilax china planting substrate and a preparation method thereof

Abstract

The application discloses a yunnan rhizoma polygonati planting substrate and a preparation method thereof, relates to the technical field of Chinese herbal medicine planting, and the substrate comprises a base layer, a core layer and a surface covering layer. The core layer comprises specific medicinal plant residue fermentation products, is prepared by specific pretreatment processes of specific plant residues through multi-stage fermentation and sterilization of a compound microbial agent, is used for simulating under-forest litter and releasing substances capable of inducing physiological activity of yunnan rhizoma polygonati, and adds a core microbial component, at least including an arbuscular mycorrhizal fungus capable of constructing and maintaining a loose, breathable and porous structure of the core layer and enhancing nutrient absorption, and an efficient root zone nitrogen-fixing bacterial group. The surface covering layer is a lichen symbiotic nitrogen-fixing cyanobacterium, which cooperates with the efficient root zone nitrogen-fixing bacterial group of the core layer, provides double biological nitrogen fixation sources of a surface layer and a root zone for the yunnan rhizoma polygonati, and the substrate can significantly promote healthy growth of the yunnan rhizoma polygonati, improves soil structure, enhances plant stress resistance and nutrient utilization efficiency, and has good ecological benefits.

Description

Technical Field

[0001] This invention relates to the field of traditional Chinese medicine cultivation technology, specifically a cultivation substrate for Polygonatum yunnanense and its preparation method. Background Technology

[0002] As an important traditional Chinese medicine, Polygonatum yunnanense has high medicinal and economic value. With the increase in market demand, higher requirements have been placed on the standardized and ecological cultivation techniques of Polygonatum yunnanense. At present, the cultivation of Polygonatum yunnanense still faces many challenges. For example, how to construct a planting substrate that can meet its specific nutritional needs, promote its healthy growth, improve the quality of the medicinal material, enhance the plant's resistance to stress, reduce the occurrence of pests and diseases, and maintain the long-term health of the soil is a technical problem that urgently needs to be solved in this field. Traditional planting methods often rely on experience and lack standardized substrate formulas that can make full use of modern biotechnology and ecological principles. This may lead to poor plant growth, frequent diseases, and inconsistent quality of medicinal materials. Moreover, excessive reliance on exogenous fertilizers and pesticides also brings hidden dangers to environmental and quality safety.

[0003] There have been some attempts in the prior art to develop cultivation substrates or methods for Polygonatum plants. For example, Chinese invention patent CN115024185B discloses a substrate for the artificial cultivation of Polygonatum multiflorum, which is mainly composed of magnesium-modified attapulgite, peat moss, wood ash, neem bark, and mulberry leaves. It aims to prevent pests and reduce pesticide use by using specific additives (such as neem bark and mulberry leaves). This scheme has made some explorations in the use of natural substances for pest and disease prevention. Another Chinese invention patent, CN115362897B, discloses a cultivation method for Polygonatum that helps improve the quality of Polygonatum. This method involves rapid propagation by tissue culture and a cultivation substrate containing peat moss, wood ash, and a specific fermentation product (a mixture of sweet potato vines, pine needles, corn stalks, and EM bacterial solution for fermentation). It is used in conjunction with a culture medium containing Bacillus thuringiensis and Bacillus coagulans to improve the transplant survival rate and disease resistance of tissue culture seedlings. This scheme focuses on the early growth and adaptability of tissue culture seedlings and introduces microorganisms and fermentation products.

[0004] The above designs aim to improve plant growth, prevent and control pests and diseases, or increase transplant survival rate by introducing specific natural materials or microorganisms, but they still have certain limitations: the integrity and synergy of the ecosystem construction are insufficient, the potential for bioactivity and functional induction of the substrate is not explored in depth, the refined design of substrate structure and functional zoning is lacking, and there is insufficient systematic consideration of meeting the specific physiological needs of Polygonatum yunnanensis and improving the quality of medicinal materials.

[0005] Therefore, there is an urgent need for a special ecological planting substrate for Polygonatum yunnanensis and its preparation method that can overcome the above limitations by constructing a reasonable structure, optimized components, especially including fermented medicinal plant residues prepared through a specific pretreatment process to simulate natural habitats and induce plant physiological activities, while integrating a complex microbial community including arbuscular mycorrhizal fungi and a highly efficient dual biological nitrogen fixation system, so as to achieve efficient nutrient circulation, rhizosphere microecological balance, healthy plant growth and improved quality of medicinal materials. Summary of the Invention

[0006] The purpose of this invention is to overcome the shortcomings of the prior art and propose a planting substrate for Polygonatum yunnanense and its preparation method to solve the above-mentioned problems.

[0007] The purpose of this invention is achieved through the following technical solution: a planting substrate for Polygonatum yunnanense and its preparation method, comprising, arranged from bottom to top, the following components occupying a predetermined volume ratio of the total substrate: a base layer (10); a core layer (20), the core layer (20) occupying 60-75% of the total volume of the substrate, comprising: (I) a physical substrate, the physical substrate being composed of the following three components, and the sum of the volume percentages of these three components being 100%: (A) a specific medicinal plant residue fermentation product, occupying 5-10% of the total volume of the physical substrate of the core layer, the fermentation product being obtained by using a compound microbial agent containing at least one of EM bacteria, brewer's yeast, Lactobacillus plantarum or Trichoderma viride for multi-stage fermentation under controlled temperature and humidity and then being treated by pasteurization or high-temperature steam sterilization process, the fermentation product being used to simulate the characteristics of forest litter and to release secondary metabolites that mildly stimulate the growth of Polygonatum yunnanense or its physiological activity;

[0008] (B) Functional composite additive component, accounting for 5-10% of the total volume of the core layer physical matrix, the functional composite additive component itself is composed of the following sub-components mixed in approximately weight percentages: food-grade diatomaceous earth or micronized zeolite (40-50%); a 1:1 mixture of phosphate rock powder and calcined oyster shell powder or bone meal (20-30%); and high-quality humic acid or fulvic acid granules (as the remaining portion to supplement the weight of the aforementioned sub-components to 100%, usually accounting for 20-40% of the total weight of the functional composite additive component).

[0009] (C) The basic organic and inorganic mixed medium, whose volume is determined by the specific medicinal plant residue fermentation product (A) and (B) functional compound additive components, is used to supplement the remaining portion of the core layer physical matrix to 100% of the total volume. This remaining portion usually accounts for 80-90% of the total volume of the core layer physical matrix. The basic organic and inorganic mixed medium itself is composed of the following sub-components mixed in approximate volume percentages: fully fermented pine needle soil or broadleaf humus (40-50%); high-quality coconut coir (20-25%); a 1:1 mixture of perlite and vermiculite (10-15%); fine biochar powder pretreated and activated with fish protein hydrolysate or fermented soybean cake liquid (5-10%); and refined earthworm castings (as the remaining portion supplementing the aforementioned sub-components to 100% of the volume, usually accounting for 5-20% of the total volume of the basic organic and inorganic mixed medium).

[0010] (II) Core microbial components, added to the physical matrix, the core microbial components include at least: an arbuscular mycorrhizal fungi (AMF) agent that can effectively colonize the roots of Polygonatum yunnanensis, construct and maintain a loose and breathable porous structure in the core layer (20) through its extensive hyphal network and promote the formation of soil aggregates and enhance nutrient absorption; and a highly efficient rhizosphere nitrogen-fixing bacteria community;

[0011] A top cover layer (30) is located above the core layer (20) and has a thickness of 1-3 cm. It comprises: a base cover material (e) composed of fine-grained Akadama soil or Kanuma soil mixed with weathered pine bark fragments or coarse coconut coir powder in a volume ratio of 1:1 to 1:2, which has a micro-topography with small undulations created by human intervention; a second microbial component (f) which comprises dried fragments or powdery buds of lichens containing nitrogen-fixing cyanobacteria selected from the genus *Lithocarpus* or suitable *Lithocarpus*, and is spread at a rate of 1-5 grams of dried lichen fragments or powdery buds per square meter of the top cover layer surface, and is mixed with diluted milk liquid, rice water or 0.1-0.5% (w / v) sodium alginate solution as an adhesion and germination promoter; wherein, the nitrogen-fixing cyanobacteria symbiotic with lichens in the top cover layer (30) and the highly efficient rhizosphere nitrogen-fixing bacteria in the core layer (20) work synergistically to provide a dual biological nitrogen fixation source for *Polygonatum yunnanense* in the surface and rhizosphere.

[0012] The base layer (10) comprises 15-20% of the total volume of the matrix and consists of the following components in approximate volume percentages: coarse river sand comprising 20-30% of the layer volume; large pieces of bamboo charcoal or hardwood charcoal comprising 10-20% of the layer volume; and at least one or a mixture thereof selected from porous volcanic rock, large-particle ceramsite, high-temperature treated crushed walnut shells or chestnut shells, as the remaining portion comprising up to 100% of the volume of the base layer (10); and the core microbial components in the core layer (20) further constitute the main body of the first microbial community, which is further supplemented with: at least one saprophytic fungal agent selected from the Trichoderma genus, with an inoculation amount of 1 x 10⁻¹⁰ per gram of physical matrix of the core layer. 5 -1x10 7 CFU; The highly efficient rhizosphere nitrogen-fixing bacterial community consists of at least one bacterium selected from Azotobacter paspalumana, Azotobacter brevis, or nitrogen-fixing Bacillus, with a total nitrogen-fixing bacteria inoculum of 1 x 10⁻⁶ bacteria per gram of core layer physical substrate. 6 -1x10 8 CFU; the inoculum size of the arbuscular mycorrhizal fungi (AMF) inoculum is 50-500 viable propagules per gram of core layer physical substrate; the first microbial community also includes at least one phosphate-solubilizing bacterial inoculum selected from Bacillus megaterium or phosphate-solubilizing Pseudomonas fluorescens, with an inoculum size of 1 x 10⁻⁶ cells per gram of core layer physical substrate. 6 -1x10 8 CFU, and / or at least one potassium-solubilizing bacterial agent selected from Bacillus mucilaginosus or Bacillus mycoides, at an inoculum dosage of 1 x 10 CFU per gram of core layer physical substrate. 6 -1x10 8 CFU.

[0013] The first microbial community in the core layer (20) also contains at least one plant growth promoter (PGPR) or biocontrol microbial agent selected from Bacillus subtilis or antagonistic Streptomyces, with an inoculation amount of 1 x 10^6 micrograms per gram of physical substrate in the core layer. 6 -1x10 8 CFU.

[0014] The physical matrix (I) component (B) functional composite additive component in the core layer (20) also contains food-grade chitosan (molecular weight 5-200,000 Da) particles or seaweed extract (containing seaweed polysaccharide ≥20%) particles, accounting for 0-5% of the weight of the component.

[0015] The arbuscular mycorrhizal fungi (AMF) inoculum in the core layer (20) contains at least two different genera or species of AMF to enhance adaptability to different environmental conditions and symbiotic efficiency with Polygonatum yunnanense.

[0016] The second microbial component (f) in the mulch layer (30) also contains arbuscular mycorrhizal fungi (AMF) spores of the same or synergistic species as those used in the core layer (20), at a spreading rate of 1 x 10⁻⁶ spores per square meter of mulch surface. 3 -1x10 5 One spore.

[0017] In the core layer (20), the components (C) of the physical matrix (I) are: fully fermented pine needle soil or broadleaf humus soil accounts for 45% of its volume, high-quality coconut coir accounts for 23% of its volume, a 1:1 mixture of perlite and vermiculite accounts for 12% of its volume, pre-treated and activated fine biochar powder accounts for 10% of its volume, and refined earthworm castings account for 10% of its volume.

[0018] A method for preparing a planting substrate for Polygonatum yunnanense includes the following steps:

[0019] (S1) Preparation of the base layer (10): According to the description of the components of the base layer (10) and their volume percentage in the layer, lay up the material to form a base layer accounting for 15-20% of the total matrix volume;

[0020] (S2) Preparation of the core layer (20):

[0021] (S2a) The basic organic and inorganic mixed medium is prepared by mechanical stirring or manual mixing according to the components (C) of the physical matrix (I) of the core layer (20), the sub-components of the basic organic and inorganic mixed medium and their volume percentage in the medium.

[0022] (S2b) The basic organic and inorganic mixed medium obtained in step (S2a) and the components (A) specific medicinal plant residue fermentation product and (B) functional compound additive components of the physical matrix (I) of the core layer (20) are uniformly mixed by mechanical stirring or manual stirring according to their predetermined percentage in the total volume of the physical matrix of the core layer to form the physical matrix body of the core layer.

[0023] (S2c) The various microbial agents involved in the description of the core microbial components (II) of the core layer (20) and the supplementary description of the first microbial community are uniformly inoculated or mixed into the core layer physical matrix body prepared in step (S2b) according to their predetermined inoculation amount.

[0024] (S2d) Place the core layer matrix processed in step (S2c) on the base layer (10) to achieve a thickness of 60-75% of the total matrix volume.

[0025] (S3) Preparation of the topcoat layer (30): The basic covering material (e) component of the topcoat layer (30) is mixed in proportion and laid on the core layer (20) and the micro-topography with slight undulations is prepared. Then, the dried fragments or powdery sprouts of nitrogen-fixing cyanobacteria lichens in the second microbial component (f) of the topcoat layer (30) are mixed with the adhesion and germination promoter at a spreading rate of 1-5 grams per square meter of the topcoat surface and evenly spread or sprayed on the surface to achieve a thickness of 1-3 cm.

[0026] The preparation of the specific medicinal plant residue fermentation product of the core layer (20) physical matrix (I) component (A) includes: (A) specific plant residues, using a compound microbial agent containing at least one of EM bacteria, brewer's yeast, plant lactobacillus or green fungus, anaerobic fermentation for 7-15 days at a temperature of 25-35℃ and material humidity of 50-65%, followed by 15-30 days of aerobic composting fermentation, and after the fermentation is completed, pasteurization (temperature 65℃, lasting 30 minutes) or high temperature steam sterilization (temperature 121℃, lasting 20 minutes) is carried out.

[0027] The core microbial component and supplementary component of the first microbial community added in step (S2c), or the second microbial component (f) (containing AMF spores in addition to lichen vegetative cells) added in step (S3), are added after being mixed with a carrier material selected from calcium alginate, food-grade diatomaceous earth or pregelatinized starch at a ratio of 1-10% (w / w) to form sustained-release bio-inducible capsules or pellets with a diameter of 0.5-5 mm. The amount of capsules or pellets added accounts for 0.1-2% of the dry weight of the physical matrix of the core layer (20).

[0028] The beneficial effects of this invention are:

[0029] First, this invention can significantly promote the healthy growth of Polygonatum yunnanense, improve its overall vitality and production potential. By constructing a core layer containing fermented residues of specific medicinal plants, an optimized ratio of basic organic and inorganic mixed media, and functional compound additives, it provides a growth environment with excellent physical properties, balanced nutrients, and long-lasting growth for the rhizomes of Polygonatum yunnanense. In particular, the introduction and diversified application of arbuscular mycorrhizal fungi (AMF) not only greatly expands the root absorption surface area through its extensive mycelial network, enhancing the absorption of key elements such as phosphorus, but also effectively promotes the formation of soil aggregates and creates a loose and breathable rhizosphere environment, thereby ensuring the healthy development of the root system and the vigorous growth of the plant, and ultimately is expected to increase the unit yield and bioaccumulation of Polygonatum yunnanense.

[0030] Secondly, this invention aims to construct and maintain a healthy and sustainable soil micro-ecosystem, thereby improving the long-term fertility and health of the soil. The application of fermented medicinal plant residues in the core layer simulates the ecological function of the natural forest litter layer. Its slow decomposition can continuously replenish soil organic matter and may release beneficial secondary metabolites, further enriching soil biodiversity. At the same time, the synergistic effect of the complex microbial community, including saprophytic fungi, phosphate-solubilizing bacteria, and potassium-solubilizing bacteria, accelerates the mineralization of organic matter and the transformation of insoluble nutrients in the soil, forming an efficient nutrient cycling system. Substances such as glomerulonephrine secreted by arbuscular mycorrhizal fungi have a long-term effect on stabilizing and improving the soil aggregate structure, reducing the risk of soil compaction, and enhancing the self-repair and buffering capacity of the substrate.

[0031] Furthermore, this invention significantly enhances the resistance of Polygonatum yunnanense to various biotic and abiotic stresses through multiple biological and ecological mechanisms, and effectively inhibits the occurrence of soil-borne diseases. For example, the biocontrol microorganisms such as Bacillus subtilis and antagonistic Streptomyces introduced into the core layer can inhibit the growth of harmful pathogens through site competition, the production of antibacterial substances, or the induction of plant systemic resistance. The biostimulants such as chitosan or seaweed extract contained in the functional compound additives can activate the Polygonatum yunnanense's own defense system, improve its adaptability and resistance to adverse environments (such as drought and temperature fluctuations). A healthy and biodiverse rhizosphere microenvironment can itself form a natural barrier against pathogenic microorganisms, thereby reducing the use of pesticides and ensuring the ecological quality of the medicinal material.

[0032] Fourth, this invention achieves efficient biotransformation and continuous supply of key nutrients (especially nitrogen, phosphorus, and potassium) through a dual biological nitrogen fixation system and the synergistic effect of multiple microorganisms, significantly improving nutrient utilization efficiency and reducing dependence on exogenous chemical fertilizers. The nitrogen-fixing cyanobacteria in the surface layer and the highly efficient rhizosphere nitrogen-fixing bacteria in the core layer form a "top-bottom synergistic" nitrogen fixation network, continuously providing bioavailable nitrogen to the plants from the air. At the same time, the phosphorus- and potassium-solubilizing microorganisms in the core layer can activate the phosphorus and potassium elements fixed in the soil, converting them into forms that can be absorbed by the plants. This nutrient supply mode, which is mainly based on biological pathways, not only meets the nutritional needs of Polygonatum yunnanensis, but also better meets the requirements of ecological agriculture for efficient resource utilization and environmental protection.

[0033] Finally, this invention provides a solid foundation for improving the quality and standardizing the production of Polygonatum yunnanense through refined matrix component preparation processes and microbial application technologies. It also embodies the advanced concept of ecological planting. For example, the strictly controlled fermentation process of specific medicinal plant residues ensures the stability of its quality and potential physiological induction effects as an organic amendment. The application of microbial sustained-release capsule or pellet technology guarantees the survival rate, duration of efficacy, and uniformity of action of beneficial microbial agents in the matrix. The comprehensive application of these technologies not only has the potential to promote the accumulation and enhancement of effective medicinal components of Polygonatum yunnanense by improving the physiological state of plants, but also provides a replicable and scalable technical path for achieving large-scale, standardized, high-quality, and sustainable production of Polygonatum yunnanense. Attached Figure Description

[0034] Figure 1 The proportions of the present invention Figure 1 ;

[0035] Figure 2 The proportions of the present invention Figure 2 . Detailed Implementation

[0036] The technical solution of the present invention will be clearly and completely described below with reference to the embodiments. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. All other embodiments obtained by those skilled in the art based on the embodiments of the present invention without creative effort are within the scope of protection of the present invention.

[0037] It should be noted that the directional concepts of "left", "right", "up", "down", "front", "back", "inner", and "outer" in the following scheme are all relative directions, and will not be listed one by one here.

[0038] Example 1:

[0039] like Figure 1 and Figure 2 As shown, this embodiment provides a special multi-layered functional planting substrate designed for the standardized artificial cultivation of Polygonatum yunnanense. The substrate structure consists of three layers from bottom to top: a base layer (10), a core layer (20), and a surface layer (30). In a specific application case (for example, using a planting pot with an inner diameter of 30cm, a height of 25cm, a theoretical capacity of approximately 17.6L, and an actual total substrate volume of approximately 10.84L), the volume occupied by each layer and its main functional positioning are as follows:

[0040] The base layer (10) accounts for 20.3% (e.g. 2.2L) of the total actual volume of the substrate. Its main function is to ensure excellent physical drainage performance, prevent water accumulation in the roots, provide bottom aeration, and form a certain physical barrier for some large soil organisms. The core layer (20) accounts for 77.5% (e.g. 8.4L) of the total actual volume of the substrate. This layer is the main functional area for the growth of the rhizomes of Polygonatum yunnanensis, the absorption of nutrients and water, and the core biological activity. The surface layer (30) is about 2 cm thick and accounts for 2.2% (e.g. 0.24L) of the total actual volume of the substrate. Its main functions include moisturizing, reducing soil surface water evaporation, inhibiting weed growth, assisting in surface nitrogen fixation, and providing a buffered micro-ecological environment for beneficial microorganisms.

[0041] To achieve its function, the base layer (10) of this embodiment is composed of the following components in approximately volume percentage: porous volcanic rock with a particle size of 8-16 mm and a rough, porous surface accounts for 60% of the base layer volume. This material has excellent permeability and can quickly drain excess water. Coarse river sand with a particle size of 5-10 mm is natural river sand. Before use, it is repeatedly washed with clean water to remove mud and fine impurities and is fully dried for later use, accounting for 25% of the base layer volume. It is used to fill the gaps between volcanic rocks, stabilize the structure, and further assist in drainage. Large bamboo charcoal is made from mature bamboo without pesticide residues. After being carbonized at high temperature (e.g., 800-1000℃), it is broken into blocks with a length, width, and thickness of not less than 2 cm, accounting for 15% of the base layer volume. Bamboo charcoal is porous and can adsorb some impurities, regulate humidity, and provide a certain habitat for aerobic microorganisms.

[0042] During preparation, the three materials mentioned above are weighed in a clean and dry environment according to the set ratio, poured into a clean container or on the laying site, and thoroughly mixed manually or with a small mixing device (such as a cement mixer) to ensure that each component is evenly distributed. Then, the mixed base layer material is evenly spread on the bottom of the planting container or the bottom of the pre-treated planting bed, and gently tamped down to form a predetermined thickness (for example, a thickness corresponding to a 2.2L volume), ensuring that the surface is roughly flat.

[0043] The core layer (20) consists of three physical matrix components: fermented medicinal plant residues (A), functional compound additive components (B), and basic organic and inorganic mixed media (C). The sum of the volume percentages of these three components is 100%.

[0044] (A) Fermented Specific Medicinal Plant Residue (7% of the total volume of the core physical matrix): This component aims to simulate the characteristics of the forest litter layer under the natural habitat of *Polygonatum yunnanense*, and utilize its fermentation products to gently stimulate the growth or physiological activity of *Polygonatum yunnanense*. Healthy, disease-free pine needles and a small amount of honeysuckle vines (non-medicinal parts) are mixed at a weight ratio of 4:1, crushed or cut to a length of 1-3 cm, and treated with a compound microbial agent. For example, for every 100 kg of treated plant residue, a commercially available high-quality EM agent (effective viable count ≥1 x 10⁻⁶) is added. 9 3 kg of CFU / ml, 1 kg of brewer's yeast (active dry yeast), and Lactobacillus plantarum (effective live count ≥ 1 x 10⁻⁶). 10 1 kg of CFU / g microbial agent. All microbial agents are commercial products purchased from reputable microbial preparation suppliers and used according to the instructions.

[0045] Fermentation process: Thoroughly mix plant residues with a pre-calculated compound microbial agent (which can be activated with a small amount of sterile water beforehand). Gradually add clean water, strictly controlling the moisture content of the material within the range of 55-65% (it should clump together when squeezed in the hand, with water visible between the fingers but not dripping). Compactly pack the mixed material into a sealed fermentation tank or pile it up and completely cover it with double-layered black agricultural plastic film, pressing the edges firmly. Place it in an environment where the temperature can be strictly maintained at 28-32℃ (e.g., through a temperature-controlled fermentation room) for 10 days of anaerobic fermentation. After anaerobic fermentation... Move the material to a well-ventilated, clean composting area and pile it loosely (about 1-1.2 meters high and 1.5 meters wide). Maintain the material moisture content at 50-60% (spray clean water as needed if it gets dry). Turn the material thoroughly once a day at regular intervals (e.g., once in the morning and once in the evening) using a small turner or manually to ensure sufficient oxygen supply and uniform fermentation. During this stage, maintain the ambient temperature at 25-35℃. Due to microbial activity, the core temperature inside the pile can naturally rise to 55-70℃ and be maintained for at least 5-7 days, after which it will gradually decrease. The total aerobic fermentation time is 20 days.

[0046] Sterilization treatment: After fermentation and maturation are completed, in order to ensure biosafety, pasteurization (the material is kept at 65°C under moist heat for 30 minutes) or more thorough high temperature and high pressure steam sterilization (121°C, 20 minutes) is adopted. After treatment, the material is naturally cooled to room temperature for use.

[0047] (B) Functional composite additive component (7% of the total volume of the core layer physical matrix): This component is designed to supplement specific mineral elements, improve the physical and chemical properties of the matrix, and provide bioactive stimulation.

[0048] Composition and proportions (by weight): Food-grade diatomaceous earth (sieved through a 60-mesh sieve) accounts for 45%; natural phosphate rock powder (P2O5 content ≥25%, sieved through a 60-mesh sieve) and oyster shell powder (main component CaCO3, sieved through a 60-mesh sieve) calcined at high temperature (e.g., 900℃, 2 hours) and pulverized and sieved are mixed at a 1:1 weight ratio, and this mixture accounts for 30% of the total additives (i.e., phosphate rock powder 15%, calcined oyster shell powder 15%); high-quality potassium humate granules (humic acid content ≥60%, water solubility ≥90%) account for 25%.

[0049] Preparation: Weigh the above-mentioned dried components precisely according to the proportions, pour them into a clean, sealed drum mixer, and mix at low speed for 20-30 minutes to ensure that the components are evenly dispersed and there are no lumps visible to the naked eye. After mixing, seal and store for later use.

[0050] (C) Basic organic and inorganic mixed medium (accounting for 86% of the total volume of the core layer physical matrix, as the remaining portion to complete (A) and (B) to 100%):

[0051] This medium is the main body of the core layer, providing the root system with the main physical support, water and basic nutrients.

[0052] Composition and proportions (by volume): 45% is pine needle humus that has been fully fermented for more than 3 years and sieved; 23% is high-quality imported coconut coir (EC value <0.5mS / cm) that has been fully desalinated; 12% is a combination of horticultural grade perlite and vermiculite (both with a particle size of 3-6mm, premixed by volume at a ratio of 1:1); 10% is fine bamboo charcoal powder (200 mesh or finer, pre-saturated with 1% concentration of commercially available fish protein hydrolysate at a charcoal-liquid weight ratio of 1:2 for 12 hours, and then air-dried to a moisture content of about 20%); and 10% is refined and decomposed commercial earthworm castings (organic matter content ≥30%).

[0053] All raw materials must be free from obvious pests and diseases and harmful chemical residues. After accurately measuring each component according to the proportion, put them into a large stainless steel horizontal ribbon mixer in sequence and stir at medium speed for 30-40 minutes until all components are mixed evenly, with consistent color and moderate humidity.

[0054] (II) Addition of core microbial components: After the physical matrix body of the core layer is formed by fully mixing the above three types of physical matrix components (A), (B) and (C) in proportion, a screened, high-quality commercial microbial agent is uniformly inoculated into it.

[0055] Arbuscular mycorrhizal fungi (AMF) inoculants are selected from compound inoculant products containing at least two highly effective AMF species (such as *Bacillus mossiosus* and *Bacillus simianus*) with a spore density ≥1000 spores / gram. The dosage is calculated based on the standard addition of 100 effective propagules per gram of physical substrate (dry weight).

[0056] High-efficiency rhizosphere nitrogen-fixing bacteria community is selected from compound inoculants containing Azotobacter paspalumandii and Azotobacter brevicornu (total effective viable count ≥ 2 x 10⁻⁶). 9 CFU / g), adding 1×10 CFU / g of core layer physical matrix. 7 CFU total nitrogen-fixing bacteria inoculation.

[0057] Trichoderma saprophytic fungi inoculant: Select Trichoderma harzianum inoculant product (effective viable count ≥ 1 x 10⁻⁶). 8 CFU / g), adding 2×10 CFU / g of core layer physical matrix. 6 CFU dosage inoculation.

[0058] Phosphate-solubilizing bacteria inoculant selected is Bacillus megaterium (effective viable count ≥ 2 x 10⁻⁶). 9 CFU / g), adding 1×10 CFU / g of core layer physical matrix. 7 CFU dosage inoculation.

[0059] Potassium-solubilizing bacteria agent selected is Bacillus mucilaginosus agent product (effective viable count ≥ 2 x 10⁻⁶). 9 CFU / g), adding 1×10 CFU / g of core layer physical matrix. 7 CFU dosage inoculation.

[0060] Add the calculated amounts of various powdered or granular microbial agents evenly during the final mixing stage of the core layer physical matrix by spreading or injecting at multiple points, and continue mixing for 5-10 minutes to ensure dispersion. Alternatively, some or all of the microbial agents can be pre-prepared into sustained-release capsules / pills before addition. Throughout the entire operation, avoid high temperatures and direct sunlight to protect the activity of the microorganisms.

[0061] Composition and preparation of the coating layer

[0062] This layer aims to retain moisture, suppress weeds, and introduce nitrogen-fixing microorganisms to the soil surface. The base cover material (e) is made of fine-grained (2-5mm diameter) Japanese Akadama soil and weathered pine bark fragments (0.5-1cm in length) that have been screened to remove large pieces, mixed evenly at a volume ratio of 1:1.5. Before use, a small amount of clean water can be sprayed until the surface of the material is slightly damp (moisture content of about 30%).

[0063] Second microbial component (f):

[0064] Preparation of lichen propagules: Collect small quantities of Peltigera spp. lichens from an area similar to the natural habitat of Polygonatum yunnanensis and free from pollution. After collection, remove impurities and dry thoroughly in a cool, ventilated place. Gently crush the dried lichens by hand or cut them into small pieces or fragments of 2-5 mm with sterilized scissors, ensuring that the asexual reproductive structures (such as powdery buds and split buds) are included.

[0065] Spread 3 grams (in the range of 1-5 grams / square meter) of the prepared lichen fragments on the substrate surface per square meter, and mix them thoroughly with 0.3% (w / v) commercially available food-grade sodium alginate aqueous solution (or sterile milk diluted 10 times) to form a slightly sticky mixture.

[0066] After the core layer is filled and leveled, the mixed base cover material is evenly spread on its surface to a predetermined thickness of 2 cm. Then, by hand or with small tools, some undulating or dotted micro-topography of about 5-10 mm is artificially created on the surface of the cover layer. Finally, the pre-moistened lichen propagule mixture is evenly sprayed or spread on the prepared surface of the cover layer, ensuring that the lichen fragments can make good contact with the moist surface material to facilitate their recovery and colonization.

[0067] Work process

[0068] The planting substrate for Polygonatum yunnanense constructed in this embodiment has a synergistic effect among its components, creating an optimized ecological environment for the growth of Polygonatum yunnanense.

[0069] The base layer provides stable support and efficient bottom drainage channels to prevent water accumulation; the basic organic and inorganic mixed media in the core layer (especially perlite, vermiculite, coconut coir and biochar) ensures good granular structure, porosity and moderate water retention capacity, providing a water-air balanced growth space for the roots. The hyphal network of arbuscular mycorrhizal fungi further shuttles through and stabilizes these granules, maintaining and improving the loose and breathable properties of the substrate in the long term.

[0070] The earthworm castings, humus, and functional compound additives such as phosphate rock powder and calcined oyster shell powder in the core layer can slowly release nitrogen, phosphorus, potassium, calcium and various trace elements. The fermentation products of specific medicinal plant residues will also continuously release organic nutrients during the decomposition process.

[0071] The symbiotic cyanobacteria in the lichen cover fix nitrogen from the air through photosynthesis. The nitrogen they release can be absorbed by the surface roots or seep into the core layer with water. The highly efficient rhizosphere nitrogen-fixing bacteria in the core layer directly fix nitrogen in the rhizosphere, providing a more direct nitrogen source for Polygonatum yunnanense. This dual biological nitrogen fixation system, which works in synergy between the top and bottom layers, can significantly improve the bioavailability and sustainability of nitrogen supply, and reduce dependence on chemical nitrogen fertilizers.

[0072] Phosphorus-solubilizing and potassium-solubilizing bacteria in the core layer can convert phosphorus and potassium elements fixed in the soil into forms that can be absorbed by plants. The hyphae of arbuscular mycorrhizal fungi (AMF) greatly expand the absorption range of the root system, especially enhancing the absorption efficiency of poorly mobile phosphorus and certain trace elements. Humic acid can chelate mineral ions and improve their availability.

[0073] The various small-molecule organic compounds, phenols, oligosaccharides, and other secondary metabolites released during the decomposition of certain medicinal plant residues may have a mild and beneficial effect on the rooting, growth, and biosynthetic pathways of specific medicinal components in Polygonatum yunnanense.

[0074] Beneficial saprophytic fungi such as Trichoderma in the core layer can supplement biocontrol bacteria, which can inhibit the growth of potential pathogenic microorganisms in the soil through competition, antagonism, and antibiotic production mechanisms, maintain the health and balance of the rhizosphere microecology, and reduce the risk of soil-borne diseases. The Akadama soil and pine bark debris cover layer with micro-topography can effectively reduce soil moisture evaporation, buffer surface temperature changes, inhibit weed growth, and provide favorable conditions for the colonization of lichens and beneficial microorganisms.

[0075] Taking a certain Yunnan Solomon's Seal producing area (e.g., an altitude of about 2300 meters, an average annual temperature of 15℃, and an annual rainfall of 800-1000 mm) as an example, a gentle slope (slope <15°) with natural forest under-sunlight conditions was selected as a cultivation test site.

[0076] On the selected plot, planting beds are dug in an east-west direction. The beds are 1.2 meters wide, 0.4 meters deep, and the length depends on the terrain. The bottom of the beds is flat and slightly inclined to one side to facilitate drainage.

[0077] Strictly following the components, proportions, and preparation methods described in 1.1 and 1.2 of this embodiment, the matrix is ​​filled in layers from bottom to top. First, a base layer material accounting for about 18% (about 7.2 cm) of the total depth is laid and appropriately compacted. Then, a core layer matrix accounting for about 70% (about 28 cm) of the total depth, which has been uniformly mixed and inoculated with core microbial components, is filled and appropriately compacted. Finally, a surface layer material of about 2 cm thickness is evenly laid on the surface of the core layer, and the micro-topography is prepared. Then, the treated lichen propagules are sown.

[0078] Select healthy, disease-free rhizome segments of Polygonatum yunnanense with plump terminal buds (e.g., 5-8 cm in length, with 2-3 buds) as planting material. Before planting, disinfect the rhizomes by soaking them in a 0.1% potassium permanganate solution for 20 minutes. After rinsing them with clean water, dry them in a cool place. Plant the rhizomes horizontally or slightly at an angle in the core layer at a density of 20 cm × 30 cm, with the buds facing upwards. The soil covering depth should be about 5-6 cm from the bud to the surface of the covering layer.

[0079] After planting, immediately water thoroughly (to settle the roots) to fully moisten the substrate. Subsequent management mainly includes: replenishing water as needed according to weather conditions and substrate moisture, always keeping the core layer moist but not waterlogged. During the rainy season, pay attention to drainage to avoid prolonged waterlogging in the beds. If the natural forest undergrowth is insufficient or too strong, shade nets with a light transmittance of about 60-70% should be erected for adjustment. Before the surface cover forms a stable layer, any weeds that grow should be removed manually in a timely manner. In the early stage of lichen establishment (about the first 2-3 months), if there is continuous dry weather, the surface cover can be sprayed with moisture to promote lichen growth. To promote recovery and growth, consider supplementing with a small amount (e.g., 1 / 3 of the original sowing amount) of treated lichen propagules 1-2 months after planting to increase coverage. Throughout the entire growth cycle of Polygonatum yunnanense (usually harvested in 2-3 years), chemical nitrogen fertilizer should generally not be applied. In the middle and late stages of growth (such as before sprouting in the spring of the second year or during the tuber enlargement period), a small amount (e.g., 50-100 grams per square meter) of well-rotted organic phosphorus and potassium fertilizer (such as fermented bone meal, wood ash, etc.) or bio-organic fertilizer can be applied to the surface of the core layer (after removing the top cover) as appropriate, depending on the plant growth and substrate nutrient status, and then lightly covered with soil.

[0080] The *Polygonatum yunnanense* cultivated using the substrate of this embodiment was compared with a control group using conventional local planting soil (e.g., ordinary mountain yellow or red soil, improved by simple tillage and a small amount of farmyard manure). The following improvements in growth performance and indicators were observed:

[0081]

[0082]

[0083] The multi-layered structure of the substrate and the scientific ratio of its components ensure that the root zone is thoroughly drained and fully aerated, while maintaining moderate humidity and a stable temperature. This significantly reduces the risk of growth stress and root rot caused by poor soil conditions (such as compaction, waterlogging, and drought). The fermentation products of specific medicinal plant residues, while mimicking the natural understory habitat and slowly releasing nutrients, may also have a positive inducing and stimulating effect on the root growth and physiological metabolism of *Polygonatum yunnanensis*, enhancing its stress resistance and nutrient absorption capacity. Simultaneously, the "top-bottom synergy" dual biological nitrogen fixation system of the lichen nitrogen-fixing layer and the nitrogen-fixing bacteria community in the core root zone provides the plant with a continuous and stable source of nitrogen. Furthermore, the environmentally friendly nitrogen source significantly reduces reliance on chemical nitrogen fertilizers, lowering production costs and environmental pollution risks. The humus, coconut coir, and biochar components in the basic organic medium, in synergy with earthworm castings and specific fermentation products, provide an excellent carrier and sustainable carbon and energy source for the stable colonization, continuous reproduction, and functional performance of numerous beneficial functional microorganisms, including arbuscular mycorrhizal fungi (AMF), nitrogen-fixing bacteria, Trichoderma, and phosphorus- and potassium-solubilizing bacteria. The long-term construction and maintenance of soil aggregate structure by the AMF mycelial network ensures the sustained excellent physical properties of the substrate, enabling the establishment of a healthy, balanced, and diverse rhizosphere micro-ecosystem that effectively inhibits harmful pathogenic microorganisms. This system enhances the plant's overall disease resistance. Phosphorus- and potassium-solubilizing bacteria in the core layer effectively activate phosphorus and potassium elements fixed in the soil and organic matter, converting them into forms easily absorbed by the plant. The AMF mycorrhizal symbiotic system acts like a vast underground absorption network, greatly expanding the effective absorption range of the roots and significantly improving the absorption efficiency of poorly mobile mineral nutrients such as phosphorus, zinc, and copper, as well as water. The entire substrate system emphasizes the recycling of organic materials and the activation of biological potential, aligning with the development direction of resource-saving and environmentally friendly agriculture. Although the components and processes involved in this embodiment are relatively refined, all raw materials can be purchased through conventional channels or according to a defined process. The preparation, substrate formulation, layering, and planting procedures are clear and specific, providing a foundation for standardized operation and widespread application in actual production. Through the above-mentioned multi-faceted and multi-level synergistic optimization, the planting substrate constructed in this embodiment can significantly promote the vegetative and reproductive growth of Polygonatum yunnanense, increase the yield of underground tubers, and simultaneously improve the physiological health of the plants, enhance their stress resistance, and induce the effects of specific physiologically active substances. It is expected to simultaneously increase the content and accumulation of key effective components (such as polysaccharides, total saponins, flavonoids, etc.) in Polygonatum yunnanense, thereby achieving a dual improvement in yield and medicinal quality, with significant economic and socio-ecological benefits.

[0084] Example 2:

[0085] like Figure 1 and Figure 2As shown, this embodiment is based on the solid foundation of the Yunnan Polygonatum ecological synergistic matrix system constructed in Embodiment 1. By optimizing the functional gain of the physical matrix components of the core layer (20), and further enhancing the diversity and synergistic effect of its core microbial community (II) and the first microbial community as its main body, it aims to provide Yunnan Polygonatum with more precise biochemical stimulation, stronger adversity resistance, more efficient nutrient utilization and a superior rhizosphere micro-ecological environment.

[0086] The Yunnan Polygonatum planting substrate described in this embodiment has a three-layer structure (base layer (10), core layer (20), and surface layer (30)). The basic proportion of each layer in the total volume, the detailed composition of the base layer (10), the preparation and addition of specific medicinal plant residue fermentation products of the physical matrix (I) component (A) in the core layer (20), and the composition and preparation of the surface layer (30) are all described in accordance with the corresponding description in Example 1. The core improvements and detailed refinements of this embodiment are mainly reflected in the specific preferred ratio of the basic organic and inorganic mixed medium of the physical matrix (I) component (C) of the core layer (20), the enhancement of the functional compound additive component (B), and the diversity of arbuscular mycorrhizal fungi (AMF) agents in the core microbial components of the core layer (II) and the supplementation of specific plant growth promoting bacteria (PGPR) or biocontrol microbial agents in the first microbial community.

[0087] To further optimize the physicochemical properties of the core layer, in this embodiment, the core layer physical matrix (I) component (C) of the basic organic and inorganic mixed medium adopts the following precise volume percentage formula (the sum of the volume percentages of each sub-component is 100%): 45% fully fermented and sieved pine needle humus; 23% high-quality desalinated coconut coir (EC value <0.5mS / cm); 12% horticultural grade perlite and vermiculite (both premixed in a 1:1 volume ratio, particle size 3-5 mm); 10% fine bamboo charcoal powder (above 200 mesh) saturated with 1% fish protein hydrolysate and air-dried for activation; and 10% refined and decomposed commercial earthworm castings (organic matter content ≥30%). This ratio aims to provide a more ideal porosity, water retention, aeration, and slow-release nutrient base.

[0088] (B) Enhancement of Functional Composite Additive Components: Based on the functional composite additive components described in Example 1 (containing food-grade diatomaceous earth or micronized zeolite, a mixture of phosphate rock powder and calcined oyster shell powder / bone meal, and high-quality humic acid or fulvic acid granules), this example further introduces a bioactive stimulant to enhance the physiological functions and stress resistance of Polygonatum yunnanense. Specifically, 3% of food-grade chitosan granules (commercially available high-quality products with a molecular weight of 100,000-150,000 Da and a degree of deacetylation ≥85%) are added to the total weight of the functional composite additive components. When preparing the functional composite additive components, these chitosan granules are pre-mixed thoroughly and uniformly with other dry powder materials (diatomaceous earth, phosphate rock powder, etc.). Then, this enhanced functional composite additive component is mixed into the overall physical matrix of the core layer at a ratio of 5-10% of the total volume of the core layer physical matrix (for example, 7% is selected in this example).

[0089] Based on the core microbial components and the first microbial community described in Example 1, this example incorporates the following functional enhancements: Enhancement of the diversity of arbuscular mycorrhizal fungi (AMF) inoculants:

[0090] The AMF inoculant used in this embodiment particularly emphasizes the diversity of strains, selecting AMF containing at least two different genera or species (e.g., a compound inoculant product containing both commercially available *Cyclocarya mosierifolia* and *Cyclocarya surae*, or *Cyclocarya stolonifera* and *Cyclocarya melanothera*) to ensure that the AMF in the inoculant can more broadly adapt to the possible microenvironmental differences in the core layer substrate and increase the probability of successfully establishing efficient symbiosis with specific *Polygonatum yunnanense* genotypes. The total effective propagule inoculation amount of AMF is still maintained in the range of 50-500 per gram of core layer physical substrate (e.g., 200 effective propagules per gram in this embodiment). These AMF inoculants are all commercially available products purchased from professional microbiology companies to ensure the purity and activity of the strains.

[0091] Targeted addition of plant growth promoters (PGPR) and biocontrol agents:

[0092] To further enhance the growth vigor of *Polygonatum yunnanense* and strengthen its resistance to soil-borne diseases, this embodiment supplements the first microbial community of the core layer (20) with a specific PGPR or biocontrol microbial agent. Specifically, *Bacillus subtilis* can be added at a rate of 1 x 10⁻⁶ per gram of physical substrate in the core layer. 7 Inoculate with CFU, and / or use antagonistic Streptomyces, adding 1x10 CFU per gram of core layer physical substrate. 7For CFU inoculation, these microbial agents are all commercially available products. In practice, one can be selected based on the expected main efficacy (focusing on growth promotion or focusing on the biological control of specific diseases), or, provided that there is no antagonistic interaction between the selected strains and their mechanisms of action are complementary, both (e.g., Bacillus subtilis and antagonistic Streptomyces) can be inoculated at half the recommended dosage (i.e., 5 x 10^6 CFU for each type). 6 The microbial agents were combined with other microbial components in the core layer physical matrix to obtain a wider range of biological effects.

[0093] The basic organic-inorganic mixed medium provides a more stable and suitable physicochemical environment for the symbiosis of roots and microorganisms, ensuring a balanced supply of water, nutrients, and air. It is the cornerstone for the efficient functioning of other bioactive components. Chitosan, as a natural biological signaling molecule, can be recognized by *Polygonatum sibiricum* cells, thereby activating defense signaling pathways within the plant (such as the jasmonic acid pathway and salicylic acid pathway), inducing the production of a series of defense-related enzymes and secondary metabolites (such as phytoalexins and PR proteins), enhancing the plant's resistance to pathogen infection (i.e., inducing systemic resistance (SAR) or systemic acquired resistance (ISR)). Simultaneously, chitosan itself and its degradation products (chitosan oligosaccharides) can directly inhibit the growth of some pathogenic fungi and bacteria, and promote plant root development and the proliferation of beneficial microorganisms (such as actinomycetes). Different species of AMF mycorrhizal fungi show preferences for root colonization, hyphal expansion range, and resistance to different pathogens. Differences may exist in the utilization capacity of speciation nutrients (especially organophosphates) and the specific mechanisms by which plant stress resistance is induced. The use of diverse AMF inoculants can improve the overall success rate and coverage of mycorrhizal colonization, enabling Polygonatum yunnanense to absorb nutrients and water from a wider range of soil microzones and potentially activate more diverse plant physiological responses, thus exhibiting greater resilience and adaptability in the face of complex environmental stresses. Bacillus subtilis and / or antagonistic Streptomyces promote the growth and development of Polygonatum yunnanense and improve nutrient availability through multiple mechanisms, such as producing plant hormones like IAA, dissolving insoluble phosphates, producing siderophores, and secreting various antibiotics and hydrolytic enzymes (such as chitinase and glucanase). They also work synergistically with AMF, Trichoderma, and other beneficial microorganisms to form a strong biological barrier in the rhizosphere, effectively competing with and antagonizing soil-borne pathogens, significantly reducing the risk of disease occurrence, and creating a healthy rhizosphere microecology.

[0094] By introducing plant immune inducers such as chitosan and highly efficient biocontrol microorganisms (such as Bacillus subtilis and antagonistic Streptomyces), a multi-layered protection system combining active defense and biological antagonism was constructed, significantly enhancing the resistance of *Polygonatum yunnanensis* to soil-borne diseases and adverse environmental stresses. The diversified application of AMF strains improved the success rate of mycorrhizal establishment and the adaptability and functional stability under different substrate microenvironments, ensuring that *Polygonatum yunnanensis* can continuously and efficiently obtain nutrients and water from the symbiotic system and achieve stronger biological protection. Specific PGPR strains not only directly promote plant growth but also improve the physicochemical properties of the rhizosphere soil, regulate root exudates, and optimize... By optimizing the composition of beneficial microorganisms, a "healthy" or "disease-suppressing" rhizosphere microecology with greater inhibitory effect on pathogens is formed. Through more refined biological regulation, the negative impacts of environmental stress and diseases on plant growth and development are reduced, which helps to achieve uniformity in the individual development of Polygonatum yunnanense. This also makes it possible to produce high-quality medicinal materials with specific high-quality characteristics (such as high content of specific medicinal components and low pesticide residues). This embodiment further reduces dependence on external chemical inputs, strengthens the self-regulation and protection functions of the substrate ecosystem, and is more in line with the concept and requirements of organic agriculture and sustainable Chinese medicinal material production, with better ecological benefits and industrial promotion prospects.

[0095] In summary, based on Example 1, Example 2 constructs a more powerful and precise micro-ecosystem for the cultivation of Polygonatum yunnanense by optimizing and enhancing specific components of the core layer matrix and targeting the enhancement of microbial community diversity and function. This results in a more significant comprehensive advantage in improving plant health, stress resistance, nutrient utilization efficiency, and the final quality of the medicinal material.

[0096] Example 3:

[0097] like Figure 1 and Figure 2 As shown, this embodiment focuses on the integrated application of bio-enhanced surface layer, refined preparation process of specific organic materials, and efficient microbial sustained-release technology. By inoculating the surface layer (30) with additional arbuscular mycorrhizal fungi (AMF), adopting a more refined and standardized preparation process for the key specific medicinal plant residue fermentation product (A) in the core layer (20), and using advanced sustained-release encapsulation technology for the microbial agents in the core layer and part of the surface layer, the overall performance and stability of the matrix system are further improved.

[0098] The preparation principles of the Yunnan Polygonatum planting substrate described in this embodiment are similar to those of Examples 1 and 2. The overall three-layer structure (base layer (10), core layer (20), and surface layer (30)) and the basic proportion of each layer in the total volume, the composition of the base layer (10), the components of the physical matrix (I) in the core layer (20), the functional composite additive components (which may include chitosan or seaweed extract as described in Example 2) and the basic organic and inorganic mixed medium (which may adopt the specific preferred ratio described in Example 2) are similar to those of Examples 1 and 2. The main improvements in this embodiment are reflected in the following aspects:

[0099] Based on the preparation of the topcoat (30) described in Example 1 (including the basic covering material (e) and the second microbial component (f) containing nitrogen-fixing cyanobacteria lichen), this example further adds arbuscular mycorrhizal fungi (AMF) spores of the same species or with good synergistic effects as those used in the core layer (20) to the second microbial component (f).

[0100] Select the same or contain species that can adapt to the surface microenvironment (such as some small root canal stalk cysts) as the core layer AMF inoculant (such as the compound inoculant of *M. mossiosus* and *M. margaritifera* mentioned in Example 2).

[0101] AMF spore dispersal rate should be controlled at 1 x 10 spores per square meter of surface mulch. 4 1 x 10 spores (in 1 x 10 3 -1x10 5 Within the recommended range of spores per square meter, these AMF spores can be mixed with dried fragments or powdery buds of lichen (1-5 grams per square meter) into the aforementioned adhesion and germination promoter (such as 0.3% w / v sodium alginate solution or diluted milk solution) to form a mixed suspension or moist powder, which is then evenly sprayed or spread on the base cover material (e) that has been laid and prepared with micro-topography.

[0102] The core layer (20) physical matrix (I) component (A) is a standardized and refined preparation process for specific medicinal plant residue fermentation products. To ensure the uniformity of quality and the maximization of bioactivity of specific medicinal plant residue fermentation products, this embodiment adopts a more rigorous and refined preparation process:

[0103] Selected pine needles (70% by weight) free from mold and pollution and dried honeysuckle vines (non-medicinal parts, 30% by weight) are used as mixed plant residues. These residues are then processed into uniform small segments of 1.0-2.5 cm in length using a chopper or pulverizer.

[0104] For every 100 kg of treated plant residue, precisely add a compound microbial fermentation agent, such as: commercially available high-activity EM bacteria stock solution (total effective live bacteria count ≥ 1 x 10⁻⁶). 100.2 liters of CFU / ml, 50 grams of brewer's yeast (active dry yeast), and Lactobacillus plantarum (effective live count ≥1x10⁻⁶). 10 50 grams of CFU / g bacterial powder can be added, along with 2-3% rice bran or corn flour (based on the dry weight of the plant residue) to adjust the initial carbon-nitrogen ratio and provide additional nutrition.

[0105] The plant residues, compound microbial agent and excipients are thoroughly mixed in a clean mixing device. Clean water is slowly added, and the moisture content of the material is monitored and precisely adjusted to 60% ± 2% using a humidity meter.

[0106] The mixed material is compacted and packed into a sealable food-grade plastic fermentation tank or a fermentation pool lined with a thick plastic film. The air is completely removed and the tank is sealed. The tank is then placed in a constant-temperature fermentation chamber where the temperature can be actively controlled at 30℃±2℃ for anaerobic fermentation for 12 days (within the range of 7-15 days). During this period, the sealing condition is checked daily to ensure anaerobic conditions.

[0107] After anaerobic fermentation, the material is quickly transferred to a clean aerobic fermentation tank or composting site with forced ventilation or easy turning. The pile is loosely stacked, with an initial height not exceeding 1.2 meters. The internal temperature and humidity of the pile are monitored in real time using temperature and humidity sensors. In the first 3-5 days, forced ventilation or turning the pile at least twice a day promotes rapid temperature rise, ensuring that the temperature at the center of the pile reaches and is maintained at the thermophilic stage of 55-65℃ for at least 7 days to fully kill pathogens, insect eggs, and weed seeds. Afterward, the turning frequency (e.g., once every 2-3 days) and water spraying volume are adjusted according to temperature and humidity changes to keep the material humidity within the range of 50-60% until the pile temperature stabilizes at 30-40℃, the material is dark brown, loosely structured, and odorless, reaching a fully decomposed state. The entire aerobic fermentation stage lasts about 20-25 days (within the range of 15-30 days).

[0108] After the fermented and decomposed material is slightly spread out, it is sent to a high-pressure steam sterilizer and thoroughly sterilized at 121°C for 30 minutes. After sterilization, it is cooled in a clean environment and sieved (e.g., a 5mm sieve) to remove large pieces of undecomposed material, resulting in a homogeneous finished fermented product, which is then sealed and packaged for later use.

[0109] The application of bio-induced capsules / granules, a highly efficient sustained-release delivery technology for microbial agents, aims to improve the survival rate, duration of effectiveness, and uniformity of distribution in the matrix of the first microbial community (including core microbial components and their supplementary components, such as AMF, nitrogen-fixing bacteria, Trichoderma, phosphate-solubilizing bacteria, potassium-solubilizing bacteria, PGPR / biocontrol bacteria, etc.) added to the core layer (20) and the AMF spores (if used) added to the outer coating layer (30). In this embodiment, a microbial sustained-release encapsulation technology is used.

[0110] Food-grade or agricultural-grade sodium alginate is selected as the main encapsulating agent, food-grade diatomaceous earth is used as the filler and drying aid, and pregelatinized starch is used as a nutritional supplement and structural modifier. For example, a 2% (w / v) sodium alginate solution can be prepared by adding 5-10% of fine diatomaceous earth powder and 1-5% of pregelatinized starch by dry weight of sodium alginate, and stirring evenly to form an encapsulation solution.

[0111] High concentrations of various microbial suspensions or spore powders are uniformly dispersed in the above-mentioned encapsulation solution at a ratio of 1-10% (w / w) of effective biomass in the final dried capsules / pills.

[0112] Using a pelletizing machine or spray granulation equipment, the embedding solution containing microorganisms is dripped into a 2-3% (w / v) calcium chloride solution and stirred rapidly to form gel microspheres with a diameter of 1-3 mm (within the range of 0.5-5 mm). The microspheres are collected, the surface calcium chloride is washed off with sterile water, and then dried to constant weight in flowing air at low temperature (e.g., 30-40°C), or freeze-drying technology is used.

[0113] When mixing the prepared sustained-release bio-inducible capsules or pellets with the core layer physical matrix, add them evenly at a rate of 0.5% (within the recommended range of 0.1-2%) of the expected dry weight of the core layer (20) physical matrix. For the AMF spores of the surface layer (if encapsulated), the encapsulated AMF spore pellets can be evenly applied together with the lichen propagules.

[0114] Inoculating the surface layer with AMF spores allows the shallow roots of *Polygonatum yunnanense* to quickly establish a symbiotic relationship with AMF from seed germination or early seedling transplantation. This not only accelerates the plant's absorption of surface water and nutrients such as nitrogen fixed by lichens, but may also lead to a closer interaction with lichens through the mycelial network, jointly improving the surface soil structure and micro-zone nutrient cycling.

[0115] Fermented medicinal plant residues prepared through a meticulously controlled multi-stage fermentation and strict sterilization process have a higher degree of organic material decomposition, and harmful organisms are completely eliminated. The types and contents of secondary metabolites (such as small molecule organic acids, enzymes, and plant growth regulator analogs) produced by beneficial microorganisms fermentation are more abundant and stable. This makes its effects in simulating the forest understory microenvironment, slowly releasing nutrients, improving soil structure, and gently inducing specific physiological activities of Polygonatum yunnanensis more significant, lasting, and predictable.

[0116] Encapsulating microorganisms in sustained-release capsules or pellets can effectively protect them from adverse environmental factors (such as dryness, ultraviolet radiation, pH fluctuations, and competition from other microorganisms) during substrate mixing, storage, transportation, and the initial application period, significantly improving their survival rate. At the same time, the sustained-release carrier can gradually release the microorganisms as the substrate humidity changes and as it degrades, ensuring a continuous and stable supply to the rhizosphere microzone during the critical period of root growth and expansion of Polygonatum yunnanense. This allows for more effective colonization and the exercise of their biological functions such as nitrogen fixation, phosphorus solubilization, potassium solubilization, growth promotion, and biocontrol, maximizing and extending the efficacy of the microorganisms.

[0117] Specific application examples

[0118] For example, an agricultural enterprise dedicated to producing GAP-standard Polygonatum odoratum medicinal materials needs to establish a highly controllable and replicable seedling and planting substrate system.

[0119] According to the refined fermentation process detailed in this embodiment, a dedicated fermentation workshop is established to carry out large-scale, standardized fermentation and sterilization of specific medicinal plant residues to ensure the quality stability of each batch of fermented products.

[0120] By collaborating with microbial technology companies or establishing small-scale packaging production lines, the selected high-efficiency AMF, nitrogen-fixing bacteria, Trichoderma, phosphorus-solubilizing and potassium-solubilizing bacteria, PGPR / biocontrol bacteria, etc., can be prepared into sustained-release bio-inducible capsules or pellets with specific bacterial load and particle size, according to the method described in this embodiment.

[0121] Using automated substrate mixing and conveying equipment, seedling trays, nutrient pots or planting troughs are prepared and filled layer by layer according to the components and precise proportions (including the amount of slow-release microbial capsules / granules) determined in Examples 1 and 2 and this Example.

[0122] After laying the base covering material on the surface of the seedling tray or planting trough, use a precision spreading device to evenly apply the pretreated nitrogen-fixing lichen propagules and the encapsulated AMF spore pellets (if used).

[0123] Because the preparation and application of key bioactive components (fermentation products, microbial agents) are highly standardized and technologically optimized, the emergence rate, uniformity, robustness and transplant survival rate of Yunnan Polygonatum seedlings are expected to be greatly improved. In subsequent growth management, due to the superior biological fertility and biocontrol capabilities of the substrate, the use of chemical fertilizers and pesticides can be significantly reduced or completely replaced, and the products are more likely to meet organic or green food standards.

[0124] The rhizosphere AMF colonization rate is higher, and the mycelial network is more developed. The activity and abundance of functional microorganisms such as nitrogen-fixing bacteria and phosphorus- and potassium-solubilizing bacteria in the core and surface layers are maintained at a high level for a long time. The physiological indicators such as chlorophyll content, photosynthetic rate, and root vigor of the plants are better, the stress response to environmental stress is smaller, and the recovery ability is stronger. The final harvested Yunnan Polygonatum tuber yield is not only high, but also more uniform in size and has good marketability. The content of the main effective components in the medicinal material (such as Yunnan Polygonatum polysaccharide, total saponins, etc.) is expected to reach or exceed the standards of high-quality medicinal materials, and the batch-to-batch differences are reduced.

[0125] Slow-release encapsulation technology ensures that beneficial microorganisms function at the most needed time and place in optimal active state, significantly improving the utilization efficiency and persistence of biological agents. This provides long-term and stable protection for the biofertility and biocontrol capabilities of the substrate. A refined and standardized preparation process is used for fermented residues of specific medicinal plants, maximizing their functions in simulating natural litter, improving soil, slowly releasing nutrients, and inducing potential physiological activity, while ensuring reliable quality. The synergistic application of nitrogen-fixing lichens and AMF spores in the surface layer not only enhances the surface's biological nitrogen-fixing capacity but also promotes early symbiosis between *Polygonatum yunnanensis* seedlings or near-surface roots and AMF, laying a solid initial foundation for rapid and healthy plant growth. Through refined control of the preparation and application techniques of key components, the entire planting substrate system exhibits more efficient and stable performance, making it easier to standardize, replicate, and scale up. It is particularly suitable for the modern Chinese medicinal herb industry, which has strict requirements for the quality of medicinal materials and the production process.

[0126] This embodiment, by integrating and applying a variety of advanced biotechnologies and ecological regulation methods, minimizes dependence on external chemical inputs and improves the utilization efficiency of agricultural resources. It represents a profound practice and beneficial exploration for realizing the ecological, intensive, and sustainable development of the Chinese medicinal herb planting industry.

[0127] Comparison of the culture medium of this invention with ordinary culture medium:

[0128]

[0129]

[0130] In summary, Example 3, through the introduction of a series of refined and integrated technologies such as AMF enhancement of the surface layer, fine fermentation process of specific organic materials, and microbial slow-release delivery, has pushed the construction concept and technical level of Yunnan Polygonatum planting substrate to a new level, providing strong technical support for achieving the production goals of Yunnan Polygonatum that are ultra-efficient, ultra-high-quality, highly ecological, and standardized.

[0131] The above description is merely a preferred embodiment of the present invention. It should be understood that the present invention is not limited to the forms disclosed herein and should not be construed as excluding other embodiments. It can be used in various other combinations, modifications, and environments, and can be modified within the scope of the concept described herein through the above teachings or related technologies or knowledge. Modifications and variations made by those skilled in the art that do not depart from the spirit and scope of the present invention should be within the protection scope of the appended claims.

Claims

1. A substrate for planting Polygonatum yunnanense, characterized in that, The substrate comprises, arranged sequentially from bottom to top, the following components occupying a predetermined volume percentage of the total substrate: a base layer (10); and a core layer (20), wherein the core layer (20) occupies 60-75% of the total volume of the substrate and comprises: (I) a physical substrate, wherein the physical substrate is composed of the following three components, and the sum of the volume percentages of these three components is 100%: (A) a fermented product of specific medicinal plant residues, accounting for 5-10% of the total volume of the physical substrate of the core layer, wherein the fermented product is prepared by using plant residues selected from the needles of pine trees and non-medicinal parts of honeysuckle vines, through a multi-stage fermentation process with temperature and humidity controlled by a compound microbial agent containing at least one of EM bacteria, brewer's yeast, plant lactobacillus or green fungus, followed by pasteurization or high-temperature steam sterilization. The fermented product is used to simulate the characteristics of forest litter and can release secondary metabolites that mildly stimulate the growth of Polygonatum yunnanensis or its physiological activity. (B) Functional composite additive component, accounting for 5-10% of the total volume of the physical matrix of the core layer, wherein the functional composite additive component itself is composed of the following sub-components mixed in approximately weight percentages: 40-50% food-grade diatomaceous earth or micronized zeolite; 20-30% a 1:1 mixture of phosphate rock powder and calcined oyster shell powder or bone meal; and high-quality humic acid or fulvic acid granules, as the remaining portion to supplement the aforementioned sub-components to 100% by weight. (C) A basic organic and inorganic mixed medium, the volume of which is the remaining portion after the components of the specific medicinal plant residue fermentation product (A) and the functional composite additive (B) are determined, is added to the total volume of the core layer physical matrix to 100%, and the remaining portion accounts for 80-90% of the total volume of the core layer physical matrix. This basic organic and inorganic mixed medium itself is composed of the following sub-components mixed in volume percentage: 40-50% fully fermented pine needle soil or broadleaf humus, 20-25% high-quality coconut coir, 10-15% a 1:1 mixture of perlite and vermiculite; and 5-10% fine biochar powder pretreated and activated with fish protein hydrolysate or fermented soybean cake liquid. And the remaining portion of refined vermicompost as a supplement to the aforementioned sub-components to bring the volume to 100%; (II) Core microbial components, added to the physical matrix, wherein the core microbial components include at least: arbuscular mycorrhizal fungi (AMF) inoculum that can effectively colonize the roots of Polygonatum yunnanensis, construct and maintain a loose and breathable porous structure in the core layer (20) through its extensive hyphal network and promote the formation of soil aggregates, and enhance nutrient absorption. And a highly efficient rhizosphere nitrogen-fixing bacterial community; A topcoat (30), located above the core layer (20), is 1-3 cm thick and comprises: a base covering material (e) composed of fine-grained Akadama soil or Kanuma soil mixed with weathered pine bark fragments or coarse coconut coir powder in a volume ratio of 1:1 to 1:2, the material having a micro-topography with slight undulations created artificially; a second microbial component (f), comprising dried fragments or powdery buds of lichens containing nitrogen-fixing cyanobacteria selected from the genus *Lithocarpus* or suitable *Lithocarpus*, the amount of which is 1-5 grams of dried lichen fragments or powdery buds per square meter of the topcoat surface, and mixed with diluted milk, rice water or 0.1-0.5% w / v sodium alginate solution as an adhesion and germination promoter; wherein, the nitrogen-fixing cyanobacteria symbiotic with lichens in the topcoat (30) and the highly efficient rhizosphere nitrogen-fixing bacteria in the core layer (20) work synergistically to provide a dual biological nitrogen fixation source for *Polygonatum yunnanense* in the surface and root zones.

2. The planting substrate for Polygonatum yunnanense according to claim 1, characterized in that: The base layer (10) comprises 15-20% of the total volume of the matrix and is composed of the following components in approximate volume percentages: coarse river sand comprising 20-30% of the layer volume; large pieces of bamboo charcoal or hardwood charcoal comprising 10-20% of the layer volume; and at least one or a mixture thereof selected from porous volcanic rock, large-particle ceramsite, high-temperature treated crushed walnut shells or chestnut shells, as the remaining portion comprising up to 100% of the volume of the base layer (10); and the core microbial components in the core layer (20) further constitute the main body of the first microbial community, which further includes: at least one saprophytic fungal agent selected from the Trichoderma genus, with an inoculation amount of 1 x 10⁻¹⁰ per gram of physical matrix of the core layer. 5 -1x10 7 CFU; The highly efficient rhizosphere nitrogen-fixing bacterial community consists of at least one bacterium selected from Azotobacter paspalumana, Azotobacter brevis, or nitrogen-fixing Bacillus, with a total nitrogen-fixing bacteria inoculation amount of 1 x 10⁻⁶ bacteria per gram of core layer physical substrate. 6 -1x10 8 CFU; the inoculation amount of the arbuscular mycorrhizal fungi (AMF) inoculum is 50-500 effective propagules per gram of core layer physical substrate; the first microbial community also contains at least one phosphate-solubilizing bacterial inoculum selected from Bacillus megaterium or phosphate-solubilizing Pseudomonas fluorescens, and the inoculation amount is 1 x 10 CFU per gram of core layer physical substrate. 6 -1x10 8 CFU, and / or at least one potassium-solubilizing bacterial agent selected from Bacillus mucilaginosus or Bacillus mycoides, at an inoculum dosage of 1 x 10 CFU per gram of core layer physical substrate. 6 -1x10 8 CFU.

3. The planting substrate for Polygonatum yunnanense according to claim 2, characterized in that: The first microbial community in the core layer (20) also contains at least one plant growth promoter (PGPR) or biocontrol microbial agent selected from Bacillus subtilis or antagonistic Streptomyces, with an inoculation amount of 1 x 10^6 micrograms per gram of physical substrate in the core layer. 6 -1x10 8 CFU.

4. The planting substrate for Polygonatum yunnanense according to claim 3, characterized in that: The physical matrix (I) component (B) functional composite additive component in the core layer (20) also includes food-grade chitosan particles with a molecular weight of 5-200,000 Da or seaweed extract particles with a seaweed polysaccharide content of ≥20%, accounting for 0-5% of the weight of the component.

5. The planting substrate for Polygonatum yunnanense according to claim 4, characterized in that: The arbuscular mycorrhizal fungi (AMF) agent in the core layer (20) contains at least two different genera or species of AMF to enhance adaptability to different environmental conditions and symbiotic efficiency with Polygonatum yunnanense.

6. The planting substrate for Polygonatum yunnanense according to claim 5, characterized in that: The second microbial component (f) in the surface layer (30) also contains arbuscular mycorrhizal fungi (AMF) spores of the same or synergistic species as those used in the core layer (20), and is sown at a rate of 1 x 10³ - 1 x 10³ spores per square meter of surface layer. 5 One spore.

7. The planting substrate for Polygonatum yunnanense according to claim 6, characterized in that: In the core layer (20), the components (C) of the physical matrix (I) are a mixture of organic and inorganic media, in which fully fermented pine needle soil or broadleaf humus accounts for 45% of its volume, high-quality coconut coir accounts for 23% of its volume, a 1:1 mixture of perlite and vermiculite accounts for 12% of its volume, pre-treated and activated fine biochar powder accounts for 10% of its volume, and refined earthworm castings account for 10% of its volume.

8. A method for preparing the *Polygonatum yunnanense* planting substrate according to claim 7, characterized in that: Includes the following steps: (S1) Preparation of the base layer (10): According to the description of the components of the base layer (10) and their volume percentage in the layer, lay up the material to form a base layer accounting for 15-20% of the total matrix volume; (S2) Preparation of the core layer (20): (S2a) The basic organic and inorganic mixed medium is prepared by mechanical stirring or manual mixing according to the components (C) of the physical matrix (I) of the core layer (20), the sub-components of the basic organic and inorganic mixed medium and their volume percentage in the medium. (S2b) The basic organic and inorganic mixed medium obtained in step (S2a) and the components (A) specific medicinal plant residue fermentation product and (B) functional compound additive components of the physical matrix (I) of the core layer (20) are uniformly mixed by mechanical stirring or manual stirring according to their predetermined percentage in the total volume of the physical matrix of the core layer to form the physical matrix body of the core layer. (S2c) The various microbial agents involved in the description of the core microbial components (II) of the core layer (20) and the supplementary description of the first microbial community are uniformly inoculated or mixed into the core layer physical matrix body prepared in step (S2b) according to their predetermined inoculation amount. (S2d) Place the core layer matrix processed in step (S2c) on the base layer (10) to achieve a thickness of 60-75% of the total matrix volume; (S3) Preparation of the topcoat (30): The topcoat (30) is made by mixing the materials of the basic covering material (e) component in the topcoat (30) in proportion and laying it on the core layer (20) and shaping the micro-topography with slight undulations. Then, the dried fragments or powdery buds of nitrogen-fixing cyanobacteria lichens in the second microbial component (f) of the topcoat (30) are mixed with the adhesion and germination promoter at a spreading rate of 1-5 grams per square meter of the topcoat surface and evenly spread or sprayed on the surface to achieve a thickness of 1-3 cm.

9. The preparation method according to claim 8, characterized in that: The preparation of the specific medicinal plant residue fermentation product of component (A) of the physical matrix (I) of the core layer (20) includes: the specific plant residue of component (A) is subjected to anaerobic fermentation for 7-15 days using a compound microbial agent containing at least one of EM bacteria, brewer's yeast, Lactobacillus plantarum or Trichoderma viride, at a temperature of 25-35°C and a material humidity of 50-65%, followed by 15-30 days of aerobic composting fermentation, and after the fermentation is completed, pasteurization is performed at a temperature of 65°C for 30 minutes, or high-temperature steam sterilization is performed at a temperature of 121°C for 20 minutes.

10. The preparation method according to claim 8, characterized in that: The core microbial component and supplementary component of the first microbial community added in step (S2c), or the second microbial component (f) added in step (S3), which, in addition to lichen propagules, contains AMF spores, are added after being mixed with a carrier material selected from calcium alginate, food-grade diatomaceous earth or pregelatinized starch at a ratio of 1-10% w / w to form a slow-release bio-inducible capsule or pellet with a diameter of 0.5-5 mm. The amount of the capsule or pellet added accounts for 0.1-2% of the dry weight of the physical matrix of the core layer (20).

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