Preparation method and application of radix peucedani seminal root germination accelerating and rooting liquid

Abstract

The invention relates to a preparation method of a peucedanum praeruptorum seminal root germination accelerating and rooting solution, which comprises the following steps: S1, raw material treatment: collecting willow twigs, peeling off the fresh barks of the willow twigs, cleaning, and cutting into small sections; s2, fermentation treatment: mixing the willow twig fresh bark segments with water, adding a fermentation aid, adjusting the pH value to be acidic, and fermenting at 25-35 DEG C to obtain fermentation liquor; the fermentation auxiliary agent is a microbial agent and / or an enzyme preparation capable of decomposing plant cell walls; and S3, post-treatment: carrying out solid-liquid separation on the fermentation liquid to obtain the germination accelerating and rooting liquid.

Description

Technical Field

[0001] This application relates to plant cultivation technology, and more specifically, to a method for preparing and applying a root-promoting solution for the germination and rooting of Peucedanum praeruptorum seeds. Background Technology

[0002] In the cultivation practice of Angelica dahurica root cuttings, the planting environment often exhibits diverse characteristics, including different geological structures and climatic conditions. When the soil is in a prolonged drought or has low moisture content, directly planting untreated Angelica dahurica roots presents significant challenges. The roots struggle to effectively absorb water and nutrients in dry soil, hindering germination, resulting in weak seedling growth, and even root rot or death, thus drastically reducing the overall survival rate. Furthermore, untreated roots have limited adaptability to harsh environments and are easily affected by external factors, further exacerbating planting risks. Therefore, scientifically germinating Angelica dahurica roots before planting is a crucial step in ensuring planting stability and improving production efficiency. Existing germination methods largely rely on traditional techniques, but these often suffer from complex operations and inconsistent results, failing to meet the practical needs of different geological environments.

[0003] To address the aforementioned issues, existing technologies urgently need improvement. Summary of the Invention

[0004] The purpose of this application is to provide a method for preparing and applying a root-promoting solution for Peucedanum praeruptorum seeds, which has the advantages of improving the germination rate and survival rate of the seeds in arid or low-moisture soils, reducing the risk of rotting and death, and enhancing environmental adaptability.

[0005] This application provides a method for preparing a root-promoting and germination solution for Peucedanum praeruptorum seeds, the technical solution of which is as follows: Includes the following steps: S1. Raw material processing: Collect tender willow branches, peel off their fresh bark, wash them, and cut them into small sections; S2. Fermentation treatment: Mix small pieces of fresh bark from tender willow branches with water, add fermentation aids, adjust the pH to acidic, and ferment at 25-35℃ to obtain fermentation liquid; the fermentation aids are microbial agents and / or enzyme preparations that can decompose plant cell walls. S3. Post-treatment: The fermentation broth is subjected to solid-liquid separation to obtain a sprouting and rooting solution.

[0006] Furthermore, this application also proposes that, in step S2, the pH value be adjusted to 4-6.

[0007] Furthermore, this application also proposes that, in step S2, the fermentation temperature is 28-32°C.

[0008] Furthermore, this application also proposes that, in step S2, the microbial agent includes at least one of Aspergillus niger, Trichoderma, and Lactobacillus plantarum.

[0009] Furthermore, this application also proposes that the microbial agent is a compound microbial agent containing Aspergillus niger, Trichoderma and Lactobacillus plantarum, and the inoculum amount is 0.1%-0.5% of the total mass of the mixed system.

[0010] Furthermore, this application also proposes that, in step S2, the enzyme preparation includes cellulase and pectinase; the fermentation treatment specifically includes: first adding the enzyme preparation for enzymatic hydrolysis, and then adding the microbial agent for fermentation.

[0011] Furthermore, this application also proposes that the enzymatic hydrolysis conditions are: temperature 40-50℃, time 2-4 hours; and the total amount of enzyme preparation added is 0.05%-0.2% of the total mass of the mixed system.

[0012] Furthermore, this application also proposes that, in step S3, after solid-liquid separation, there are further steps of concentration and / or addition of preservatives.

[0013] Furthermore, this application also proposes the application of the germination and rooting solution prepared by the above method in promoting the germination and / or rooting of Peucedanum praeruptorum seeds.

[0014] Furthermore, this application also proposes that the method for operating the above application includes: soaking the roots of Peucedanum praeruptorum in the undiluted or diluted rooting solution for 1-2 hours before planting; or, germinating the soaked roots of Peucedanum praeruptorum in the dark at 15-25℃ for 2-3 days before planting.

[0015] As can be seen from the above, the preparation method and application of the root-promoting and germination solution of Peucedanum praeruptorum provided in this application include raw material processing, fermentation processing and post-processing steps. By preparing the root-promoting and germination solution, the root germination and rooting of the seed are promoted, and the survival rate of planting in arid or low-moisture soil is improved. It has the advantages of improving the germination rate and survival rate of the seed roots in arid or low-moisture soil, reducing the risk of rot and death, and enhancing environmental adaptability. Attached Figure Description

[0016] Figure 1 This is a flowchart illustrating the preparation method of the root-promoting and germination solution for Peucedanum praeruptorum in this application embodiment.

[0017] Figure 2 This is a diagram of the budding sample of the root of *Peucedanum praeruptorum* after treatment with germination solution in an embodiment of this application. Detailed Implementation

[0018] To make the objectives, technical solutions, and advantages of the embodiments of this application clearer, the technical solutions of the embodiments of this application will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some embodiments of this application, not all embodiments. Based on the embodiments of this application, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of this application.

[0019] The following disclosure provides numerous different embodiments or examples for implementing various structures of the invention. To simplify the disclosure, specific examples of components and arrangements are described below. These are merely examples and are not intended to limit the scope of the invention. Furthermore, reference numerals and / or letters may be repeated in different examples. Such repetition is for simplification and clarity and does not in itself indicate a relationship between the various embodiments and / or arrangements discussed.

[0020] Traditional methods for preserving Peucedanum praeruptorum seeds often involve sand storage or cellar stacking. However, improper control of humidity and temperature during storage frequently leads to root rot, mold growth, or premature sprouting, resulting in decreased root viability or even rendering the seeds unusable. These methods lack precise control over humidity and ventilation, making them prone to mold growth. Furthermore, excessively thick stacking can cause internal heating and rotting, making it difficult to maximize the preservation of the root's physiological activity.

[0021] In the cultivation and production of root-propagated Angelica dahurica, the traditional existing planting methods, especially when the roots are not treated to promote germination, significantly increase the planting risk when faced with prolonged drought and dry soil conditions, affecting the survival rate and yield of Angelica dahurica.

[0022] In response, this application proposes a method for preparing a root-promoting and germination-promoting solution for Peucedanum praeruptorum seeds. This method involves collecting tender willow branches and processing their fresh bark, followed by fermentation to obtain active substances. Finally, the root-promoting solution is obtained through solid-liquid separation. The aim is to effectively promote the germination and rooting of Peucedanum praeruptorum seeds, thereby reducing the planting risk under adverse environments.

[0023] For ease of understanding, the following explains some key terms in this embodiment: Peucedanum praeruptorum root sprouting and rooting solution: refers to a liquid used to treat Peucedanum praeruptorum roots. Its main function is to stimulate the dormancy state of the roots, promote the rapid germination of buds and the growth of new roots, so as to improve the survival rate of planting.

[0024] Fresh bark from tender willow branches: refers to the fresh bark peeled off from newly grown branches of willow trees. It is believed to contain natural plant growth regulators and is the source of active ingredients for preparing bud-promoting and root-promoting solutions.

[0025] Fermentation aids: refer to substances added during the fermentation process that can promote or accelerate the fermentation reaction. In this application, they specifically refer to microbial agents and / or enzyme preparations that can decompose plant cell walls to help release the effective components in fresh willow bark.

[0026] Microbial agents: refer to preparations containing specific microorganisms that can decompose complex organic matter through their metabolic activities. In this application, they are used to decompose the cell wall structure of fresh willow bark.

[0027] Enzyme preparations: refer to preparations containing specific biological enzymes that can catalyze specific biochemical reactions. In this application, they are used to hydrolyze cell wall components such as cellulose and pectin in fresh willow bark.

[0028] This application provides a method for preparing a root-promoting and germination solution from Peucedanum praeruptorum seeds, which includes the following steps: In the raw material processing step, tender branches can be collected from naturally growing willow trees or from specially cultivated willow groves. The bark of the tender branches can be peeled off by hand or with the assistance of simple mechanical scraping tools. The peeled bark can be rinsed in clean water to remove surface impurities, and then cut into small pieces several centimeters in length using a hand knife or simple chopping equipment to increase the surface area for subsequent processing.

[0029] In the fermentation process, the cut pieces of fresh willow bark can be mixed with an appropriate amount of water to form a mixture. Fermentation aids can be a single microbial agent, such as common saprophytic bacteria, or a single enzyme preparation, such as commercially available crude enzymes, or a simple mixture of both. The pH of the fermentation system can be adjusted to the acidic range by adding common acidic substances, such as acetic acid or citric acid, and monitoring with pH test paper or a simple pH meter. Subsequently, the mixture is placed in a container capable of maintaining a temperature, such as an insulated container with a heating rod, to keep the system temperature between 25°C and 35°C for a period of time until a fermentation broth is obtained.

[0030] In the post-processing step, the obtained fermentation broth can be treated using simple solid-liquid separation methods. For example, multiple layers of gauze or a filter screen can be used to separate the liquid portion from the solid residue. The separated liquid is the initially obtained germination and rooting solution, which can be directly used for subsequent applications.

[0031] The preparation method proposed in this application utilizes natural growth-promoting substances from the fresh bark of tender willow branches, combined with the decomposition effect of fermentation aids, to efficiently extract and enrich active ingredients that promote the germination and rooting of Peucedanum praeruptorum seeds. The resulting germination and rooting solution effectively stimulates the activity of Peucedanum praeruptorum seeds, accelerating their germination and rooting process, thereby significantly reducing the risks under adverse planting conditions such as drought and soil dryness, and improving the survival rate and initial growth vigor of Peucedanum praeruptorum.

[0032] In some embodiments described above, adjusting the pH value to acidic levels is proposed to optimize the fermentation process. However, during implementation, the uncertainty of the pH range may lead to low fermentation efficiency or unstable results. Therefore, this application further proposes adjusting the pH value to 4-6 in step S2.

[0033] Specifically, pH is an indicator of the acidity or alkalinity of a solution, and its precise control is crucial for bio-fermentation processes. Adjusting the pH to 4-6 aims to provide a suitable acidic environment for the microbial agents and / or enzymes in the fermentation system, ensuring their activity and catalytic efficiency. This pH can be precisely controlled in several ways. For example, it can be adjusted by monitoring the pH of the fermentation broth in real time and adding diluted acidic solutions (such as dilute sulfuric acid, dilute hydrochloric acid, or acetic acid) or alkaline solutions (such as sodium hydroxide or potassium hydroxide solutions) as needed. Alternatively, buffering agents, such as acetate buffer or phosphate buffer, can be introduced to maintain pH stability, thus resisting the effects of acidic or alkaline metabolites that may be generated during fermentation.

[0034] The above technical solution precisely controls the pH value in fermentation step S2 within the range of 4-6, providing an optimal activity environment for fermentation aids (including microbial agents and / or enzyme preparations). This precise pH control effectively avoids the problem of inhibited microbial growth or reduced enzyme activity caused by excessive pH fluctuations, thus ensuring the stable progress of the fermentation process. Consequently, fermentation efficiency is significantly improved, and the consistency of the prepared germination and rooting solution is effectively guaranteed, solving the problems of low fermentation efficiency or unstable results.

[0035] In some of the embodiments described above in this application, fermentation temperature is proposed to regulate the fermentation process in order to optimize the preparation of rooting solution. However, in its implementation, the temperature range of 25-35°C may be too wide, leading to unstable microbial activity or fluctuations in fermentation efficiency, which in turn affects the uniformity of the composition of the fermentation solution and the consistency of the germination effect of the final rooting solution.

[0036] In this regard, this application further proposes that the fermentation temperature in step S2 is 28-32℃.

[0037] In step S2, the fermentation temperature is controlled at 28-32℃ to provide an optimized environment for the fermentation process, ensuring that the fermentation aids (microbial inoculants and / or enzyme preparations) can efficiently decompose plant cell walls. Precise temperature control is crucial for maintaining the metabolic activity of microorganisms and the catalytic efficiency of enzymes. Specifically, this can be achieved in the following ways: for example, a fermenter with a temperature control system can be used, with built-in heating and cooling devices combined with temperature sensors to monitor and adjust the temperature of the fermentation broth in real time, stabilizing it within the range of 28-32℃; alternatively, a constant-temperature water bath or incubator can be used, placing the container holding the fermentation mixture within it, to indirectly stabilize the fermentation broth temperature through precise control of the ambient temperature.

[0038] By precisely limiting the fermentation temperature to an optimized range of 28-32℃, the problem of unstable microbial activity and fluctuating fermentation efficiency caused by a wide fermentation temperature range is effectively solved. Under this temperature condition, the microbial inoculants and / or enzyme preparations can maintain optimal activity and catalytic efficiency, thereby promoting the full and stable decomposition of plant cell walls in small segments of willow tender bark. This precise temperature control avoids insufficient fermentation due to excessively low temperatures or enzyme inactivation due to excessively high temperatures, thus making the fermentation process more stable and controllable, resulting in a more uniform composition of the fermentation broth. The final germination and rooting solution exhibits a more consistent and excellent effect in promoting the germination and / or rooting of Peucedanum praeruptorum seeds. This significantly improves the stability of the preparation process and the reliability of product quality.

[0039] In some of the solutions described above in this application, microbial agents are proposed for decomposing plant cell walls. However, during implementation, the selection of microbial agents may not be optimized, leading to low fermentation efficiency or inconsistent results, affecting the quality of the final rooting solution. Therefore, this application further proposes that in the fermentation treatment step S2, the microbial agent includes at least one of Aspergillus niger, Trichoderma, and Lactobacillus plantarum.

[0040] Specifically, the microbial agent refers to a preparation containing specific microorganisms or microbial communities. Its main function is to utilize the metabolic activities of microorganisms to decompose organic matter, transform substances, or produce beneficial metabolites. In this application, the microbial agent is used to decompose the plant cell walls of the fresh bark of willow twigs to promote the release of active ingredients. It can exist in the form of liquid agents, solid agents (such as powders or granules), etc., and can be selected and formulated according to actual needs. For example, the bacterial liquid obtained from liquid fermentation can be used directly as the microbial agent, or the bacterial liquid can be concentrated and mixed with a carrier to prepare a solid agent. Among them, Aspergillus niger is a common filamentous fungus, known for its strong enzyme secretion ability, especially adept at secreting various hydrolytic enzymes such as cellulase and pectinase. In this application, Aspergillus niger is mainly used to efficiently decompose the cellulose components in the fresh bark of willow twigs, thereby destroying the plant cell wall structure and promoting the release of intracellular substances. In addition to directly using pure cultured Aspergillus niger strains as microbial agents, compound agents containing Aspergillus niger can also be used, or genetically engineered Aspergillus niger strains can be used to improve their enzyme activity. Trichoderma is another fungus widely found in nature, also capable of secreting various enzymes, especially cellulase and hemicellulase. In this application, Trichoderma works synergistically with Aspergillus niger to further decompose cellulose and hemicellulose in the fresh bark of willow twigs, enhancing the degradation efficiency of cell walls. Trichoderma inoculants can be used alone or in combination with other microbial inoculants. For example, Trichoderma reesei or Trichoderma harzianum, which have high cellulase activity, can be selected as microbial inoculants. Lactobacillus plantarum is a common lactic acid bacterium capable of lactic acid fermentation, producing organic acids such as lactic acid and acetic acid, and may produce some biologically active secondary metabolites. In this application, the addition of Lactobacillus plantarum helps to regulate the pH of the fermentation system, maintain an acidic environment, inhibit the growth of harmful microorganisms, and may produce substances beneficial to plant growth, thereby improving the quality of the fermentation broth. Lactobacillus plantarum can be added in the form of lyophilized powder or live bacterial solution. For example, selected Lactobacillus plantarum strains with good acid-producing capacity and environmental adaptability can be selected. The phrase "at least one" means that the microbial agent can be used alone from any one of Aspergillus niger, Trichoderma, or Lactobacillus plantarum, or in combination of any two or three of them. This flexibility allows for optimized selection based on the characteristics of the raw materials, fermentation conditions, and desired fermentation effects. For example, in some cases, only cell wall decomposition may be required, in which case Aspergillus niger or Trichoderma can be used alone; while when both cell wall decomposition and fermentation broth quality improvement are needed, a combination of Aspergillus niger and Lactobacillus plantarum, or a combination of Trichoderma and Lactobacillus plantarum, can be considered.

[0041] Through the above technical solution, in the fermentation treatment step S2, the microbial agent is limited to at least one of Aspergillus niger, Trichoderma, and Lactobacillus plantarum. This application can effectively solve the problem of low fermentation efficiency or inconsistent effects caused by improper selection of microbial agents. Specifically, Aspergillus niger and Trichoderma, as highly efficient cellulose and hemicellulose degrading bacteria, can work synergistically to more thoroughly decompose the plant cell walls of fresh bark of willow tender branches, promoting the release of intracellular effective components (such as auxin precursors and cytokinins). At the same time, the introduction of Lactobacillus plantarum, through its acid-producing characteristics, helps maintain the acidic environment of the fermentation system, inhibits the growth of miscellaneous bacteria, ensures the stability and safety of the fermentation process, and may produce metabolites beneficial to plant growth. This optimized combination or selection of microbial agents makes the fermentation process more efficient and controllable, thereby significantly improving the quality and stability of the prepared germination and rooting solution, providing a more reliable guarantee for the germination and rooting of Peucedanum praeruptorum seeds.

[0042] In some of the above-mentioned schemes of this application, microbial agents are proposed to decompose plant cell walls for fermentation. However, in the process of implementation, the use of a single strain may lead to insufficient decomposition efficiency, and the unclear inoculation amount affects the fermentation effect.

[0043] In this regard, this application further proposes that the microbial agent is a compound microbial agent containing Aspergillus niger, Trichoderma and Lactobacillus plantarum, and its inoculation amount is 0.1%-0.5% of the total mass of the mixed system.

[0044] Specifically, the compound microbial agent refers to a preparation composed of two or more microorganisms with different physiological functions or synergistic effects. In this application, the compound microbial agent aims to more efficiently and comprehensively decompose plant cell wall components in the fresh bark of willow tender branches through the synergistic effect of multiple microorganisms. Aspergillus niger is known for its strong cellulase and hemicellulase activities, which can effectively degrade cellulose and hemicellulose in plant cell walls; Trichoderma exhibits excellent performance in degrading cellulose and lignin, especially in assisting the decomposition of structurally complex lignin components; Lactobacillus plantarum, as a lactic acid bacterium, can produce lactic acid, lower the pH value of the fermentation system, inhibit the growth of harmful microorganisms, and may produce some small molecule organic acids, further promoting the decomposition of plant cell walls and the dissolution of effective components. In addition to the above combination, the compound microbial agent can also be composed of other microorganisms with similar decomposition capabilities, for example, it can contain other types of cellulose-decomposing bacteria, pectin-decomposing bacteria, or lignin-decomposing bacteria to meet the needs of different plant raw materials or fermentation conditions.

[0045] Meanwhile, the inoculum size refers to the amount of microbial inoculant added during fermentation, which has a crucial impact on fermentation efficiency and product quality. An appropriate inoculum size ensures a sufficient number of microorganisms in the fermentation system, thereby quickly initiating the fermentation process and maintaining high decomposition activity. If the inoculum size is too low, the number of microorganisms will be insufficient to effectively decompose the substrate, leading to a prolonged fermentation cycle and low efficiency; if the inoculum size is too high, it may cause competition for nutrients among microorganisms, preventing some microorganisms from fully functioning and increasing production costs. Controlling the inoculum size within the range of 0.1%-0.5% of the total mass of the mixed system aims to achieve the optimal balance between microbial activity and substrate decomposition efficiency. This inoculum size can be adjusted according to the actual fermenter volume, feed concentration, and inoculant activity. For example, in some cases, a more precise inoculum size can be determined by measuring the viable cell count or enzyme activity units of the inoculant to ensure optimal fermentation results.

[0046] By employing a compound microbial agent containing Aspergillus niger, Trichoderma, and Lactobacillus plantarum, and controlling its inoculum amount within the range of 0.1%-0.5% of the total mass of the mixed system, this application significantly improves the preparation efficiency and quality of the root-promoting solution for Angelica dahurica seeds. Specifically, the synergistic effect of Aspergillus niger, Trichoderma, and Lactobacillus plantarum allows for a more thorough and rapid decomposition of plant cell wall components such as cellulose, hemicellulose, and lignin in the fresh bark of willow tender branches, thereby promoting the full release and dissolution of effective components. Simultaneously, the lactic acid produced by Lactobacillus plantarum helps maintain the acidic environment of the fermentation system, further optimizing enzyme activity and inhibiting the growth of contaminating microorganisms, ensuring the stability and safety of the fermentation process. Precisely controlled inoculum amount ensures that the microbial agent can function at the optimal concentration in the fermentation system, avoiding both incomplete fermentation due to insufficient microbial quantity and resource waste and potential accumulation of metabolites due to excessive microbial quantity. This optimized combination not only improves fermentation efficiency and shortens the production cycle, but also makes the final germination and rooting solution richer in active ingredients and more stable in quality, thus showing a better effect in promoting the germination and rooting of Peucedanum praeruptorum seeds.

[0047] In some of the schemes described above in this application, fermentation aids are proposed to decompose plant cell walls and promote fermentation. However, in the process of implementation, unclear types of enzyme preparations and improper order of fermentation steps may lead to insufficient decomposition of plant materials and affect the activity of fermentation broth.

[0048] In this regard, this application further proposes that in step S2, the enzyme preparation includes cellulase and pectinase; the fermentation treatment specifically includes: first adding the enzyme preparation for enzymatic hydrolysis, and then adding the microbial agent for fermentation.

[0049] Cellulase is an enzyme that catalyzes the hydrolysis of cellulose, breaking down the cellulose components in plant cell walls. For example, this cellulase can be derived from *Trichoderma* or *Aspergillus* microorganisms, and its activity and purity can be selected according to practical application requirements. Pectinase, on the other hand, is an enzyme that catalyzes the degradation of pectin, breaking down the pectin components in plant cell walls. For example, this pectinase can be derived from *Aspergillus niger* or *Bacillus subtilis*, and its type can include polygalacturonase or pectin lyase. By using both enzymes simultaneously, the plant cell wall structure can be more comprehensively disrupted, releasing the internal active ingredients.

[0050] Enzymatic hydrolysis is the process of breaking down large molecules into smaller ones using the biocatalytic action of enzymes. In this step, the aforementioned enzyme preparation is added first to pre-decompose the cellulose and pectin in the fresh bark of willow tender branches, thereby effectively disrupting the plant cell wall structure and allowing the full release of beneficial substances from within the plant. For example, the enzymatic hydrolysis process can be carried out under suitable temperature and pH conditions to ensure maximum enzyme activity and for a certain duration to guarantee complete degradation.

[0051] After enzymatic pretreatment, microbial inoculants are added for fermentation. The aim is to utilize the metabolic activities of microorganisms to further decompose the enzymatic hydrolysis products and produce secondary metabolites with germination and rooting activity. For example, the microbial inoculant can be a single species or a compound species. Its inoculum size and fermentation conditions (such as temperature, time, and aeration rate) can be optimized according to the characteristics of the selected species to promote its growth and metabolism, thereby improving fermentation efficiency and product activity.

[0052] The above technical solution clarified the specific composition of the enzyme preparation in the fermentation aid, namely, cellulase and pectinase. These two enzymes work synergistically to efficiently decompose the main components of the cell wall of fresh bark in willow twigs, thereby releasing the active substances inside the plant more thoroughly. Simultaneously, by refining the fermentation process into a sequence of enzymatic hydrolysis followed by fermentation, pretreatment of the plant raw materials was achieved, allowing macromolecules to be initially degraded into smaller molecules, providing a more readily usable substrate for subsequent microbial fermentation. This step-by-step treatment method avoids the problem of incomplete decomposition that may occur during direct fermentation, significantly improving the metabolic efficiency of microorganisms and the content of active ingredients in the fermentation broth. Ultimately, this optimized preparation method can obtain a more active and effective germination and rooting solution, effectively solving the problems of insufficient decomposition of plant materials and affected fermentation broth activity, providing stronger support for the germination and rooting of Angelica dahurica seeds.

[0053] In some embodiments described above in this application, a step of enzymatic hydrolysis followed by fermentation is proposed to decompose plant cell walls and promote the generation of fermentation broth. However, in the implementation process, the conditions of the enzymatic hydrolysis step, such as temperature, time, and enzyme addition, are not specifically limited, which may lead to unstable enzymatic hydrolysis effect, low efficiency, or waste of resources, thereby affecting the uniformity and effectiveness of the fermentation broth.

[0054] In this regard, this application further proposes the following conditions for enzymatic hydrolysis: temperature 40-50℃, time 2-4 hours; the total amount of enzyme preparation added is 0.05%-0.2% of the total mass of the mixed system.

[0055] Specifically, enzymatic hydrolysis temperature is a key factor affecting enzyme activity. Enzymes have specific optimal temperature ranges within which their catalytic efficiency is highest. Below the optimal temperature, enzyme activity decreases; above the optimal temperature, the enzyme may undergo irreversible denaturation and become inactive. To ensure that enzyme preparations (such as cellulase and pectinase) can efficiently decompose plant cell walls, the enzymatic hydrolysis temperature needs to be controlled within a suitable range. In addition to 40-50℃, other suitable ranges such as 30-40℃ or 50-60℃ can be used, depending on the specific type and characteristics of the selected enzyme preparation, to ensure the effective action of the enzyme.

[0056] Meanwhile, the enzymatic hydrolysis time determines the extent to which the substrate and enzyme fully contact and react. Sufficient time ensures that the plant cell walls are fully decomposed, thereby releasing more active ingredients and providing ample substrate for subsequent fermentation. Too short a time may lead to incomplete decomposition, affecting fermentation efficiency and product quality; too long a time may result in energy waste or the generation of unnecessary byproducts. In addition to 2-4 hours, the enzymatic hydrolysis time can also be adjusted to 1-2 hours or 4-6 hours, depending on the type of raw material, the degree of pretreatment, the activity of the enzyme preparation, and the required degree of decomposition.

[0057] Furthermore, the amount of enzyme added directly affects the rate and efficiency of the enzymatic hydrolysis reaction. An appropriate amount of enzyme ensures effective substrate decomposition while avoiding increased costs and potential inhibitory effects from excessive addition. Too little enzyme may lead to incomplete hydrolysis, while too much may waste resources and may not significantly improve hydrolysis efficiency. Besides 0.05%-0.2%, the total amount of enzyme added can be adjusted to other proportions such as 0.01%-0.05% or 0.2%-0.5%, depending on the enzyme's activity units, substrate concentration, reaction conditions, and desired decomposition effect.

[0058] Through the above technical solution, this application synergistically optimizes the enzymatic hydrolysis effect by precisely controlling the temperature, time, and enzyme dosage during the process. Maintaining the temperature within the range of 40-50℃ fully utilizes the optimal activity range of cellulase and pectinase, accelerating the enzyme reaction rate and significantly improving the decomposition efficiency of plant cell walls. Setting the time to 2-4 hours balances the sufficiency of enzyme action with the process duration, ensuring sufficient substrate decomposition while avoiding energy waste or byproduct generation due to excessive time. Limiting the total enzyme dosage to 0.05%-0.2% of the total mass of the mixed system precisely controls the enzyme concentration, ensuring effective catalysis while preventing resource waste and potential negative impacts from excessive addition, thus optimizing overall resource utilization. This precise control of enzymatic hydrolysis conditions ensures efficient and uniform decomposition of plant cell walls, laying a solid foundation for the subsequent fermentation process. This significantly improves the uniformity and effectiveness of the fermentation broth, resulting in a more stable and superior germination and rooting-promoting solution.

[0059] In some of the embodiments described above in this application, a rooting solution obtained by solid-liquid separation is proposed for use in preparing rooting solution. However, in the process of implementation, the concentration of the rooting solution may be insufficient, resulting in poor performance, or it may be susceptible to microbial contamination and spoilage, which is not conducive to long-term storage and practical application.

[0060] To address this, this application further proposes that step S3, after solid-liquid separation, includes a concentration step and / or the addition of a preservative. The concentration step aims to increase the concentration of the effective active ingredients in the germination and rooting solution, thereby enhancing its efficacy in promoting the germination and rooting of Angelica dahurica seeds. Specifically, vacuum concentration technology can be used, evaporating water at a relatively low temperature by lowering the boiling point to reduce the loss of heat-sensitive active ingredients; alternatively, membrane separation technology, such as nanofiltration or reverse osmosis, can be used to separate water and small molecule impurities from the effective ingredients through selective membranes, thereby achieving concentration; alternatively, concentration can also be achieved through atmospheric pressure evaporation, but temperature and time must be strictly controlled to avoid degradation of active substances. The addition of a preservative aims to inhibit the growth and reproduction of microorganisms in the germination and rooting solution, preventing spoilage and thus extending the product's shelf life and ensuring its stability and safety during storage and transportation. Specifically, food-grade or agriculturally permitted chemical preservatives can be added, such as sodium benzoate and potassium sorbate. These preservatives have broad-spectrum antibacterial effects and can effectively inhibit a variety of microorganisms. Alternatively, natural preservatives can be selected, such as plant extracts (e.g., rosemary extract, tea polyphenols) or organic acids (e.g., lactic acid, citric acid). These natural components inhibit microbial growth while having a low potential impact on plant growth. Physical methods can also be considered, such as short-time high-temperature sterilization (e.g., pasteurization) after adding preservatives, to further reduce the microbial load.

[0061] Through the above technical solutions, this application effectively solves the problems of insufficient concentration and susceptibility to microbial contamination and spoilage in germination and rooting solutions. Specifically, the concentration process significantly increases the content of active ingredients in the rooting solution, allowing small amounts to achieve the desired germination and rooting effect, thereby improving the product's economy and efficiency. Simultaneously, the addition of preservatives effectively inhibits microbial growth, preventing deterioration of the rooting solution during storage and transportation due to microbial activity, ensuring product stability and long-term effectiveness. These measures work together to produce a germination and rooting solution with not only higher activity and stronger efficacy but also good storage stability, greatly facilitating the cultivation and production of Angelica dahurica roots, reducing planting risks, and increasing the germination and rooting rates of the roots.

[0062] In some of the above-mentioned schemes of this application, a rooting solution for promoting germination and rooting is proposed to promote plant growth. However, in the process of its implementation, there are problems such as unclear application methods, unstable effects, or inconvenient operation, especially under drought or dry soil conditions, regarding how to specifically apply the rooting solution to effectively promote the germination and rooting of Peucedanum praeruptorum seeds.

[0063] In this regard, this application proposes the application of a method for preparing a germination and rooting solution in promoting the germination and / or rooting of Peucedanum praeruptorum seeds.

[0064] This rooting and sprouting solution is not a commercially available generic product, but rather prepared using a specific method. Specifically, the preparation process involves fermenting fresh willow bark twigs. Through the action of microbial agents and / or enzymes, naturally occurring active ingredients in the willow bark (such as salicylic acid and auxin precursors) are extracted, transformed, and enriched, resulting in a liquid with specific biological activity. This preparation method ensures that the types and amounts of active substances in the rooting solution can specifically stimulate the plant's physiological mechanisms. For example, in addition to the aforementioned fermentation process using fresh willow bark twigs, other plant materials rich in plant growth regulators can also be used as raw materials, combined with different combinations of microbial agents or enzymes for biotransformation to obtain a liquid with similar rooting and sprouting effects.

[0065] This application refers to using the specifically prepared germination and rooting solution as a functional agent to improve the physiological state of the roots of *Peucedanum praeruptorum* seeds, enabling them to transition from dormancy to growth more quickly and effectively, and to form new root systems. Specifically, this application can be achieved in several ways. For example, the roots can be directly immersed in the germination and rooting solution, allowing the active ingredients in the solution to be fully absorbed by the roots, thereby activating their internal germination and rooting mechanisms. Alternatively, the rooting solution can be applied to the soil environment surrounding the roots through spraying or irrigation, indirectly promoting germination and rooting through the soil medium. It can also be combined with other agricultural techniques, such as soil conditioners and fertilizers, to achieve even better promotional effects.

[0066] Through the above technical solution, this application clarifies the specific use of a rooting and germination solution prepared by a specific method to promote the germination and / or rooting of Peucedanum praeruptorum seeds. This specifically prepared rooting solution, through a process (as described above) involving the fermentation of fresh bark from tender willow branches, produces bioactive components that specifically stimulate the roots of Peucedanum praeruptorum seeds. These active components effectively activate the physiological mechanisms of the roots, promoting their transition from dormancy to growth, accelerating the germination process, and stimulating the formation and growth of new roots. Especially under adverse conditions such as drought or dry soil, this rooting solution can significantly improve the germination and rooting rates of Peucedanum praeruptorum seeds, reduce planting risks, and increase survival rates and uniform emergence. By providing a clear application target and purpose, this application avoids the fluctuations in effectiveness caused by environmental factors in traditional planting, making the planting of Peucedanum praeruptorum seeds more efficient and stable.

[0067] In some of the embodiments described above in this application, the use of germination and rooting solution in promoting the germination and rooting of Peucedanum praeruptorum seeds is proposed. However, in the actual implementation process, the lack of a clear application method may lead to inconsistent germination effects or low efficiency.

[0068] In this regard, this application further proposes an application method, which includes: soaking the root of Peucedanum praeruptorum in the undiluted or diluted solution of the germination and rooting solution for 1-2 hours before planting; or, germinating the soaked root of Peucedanum praeruptorum in the dark at 15-25℃ for 2-3 days before planting.

[0069] Specifically, the soaking treatment aims to allow the roots of *Peucedanum praeruptorum* to fully absorb the active ingredients in the germination and rooting solution, thereby activating their physiological activity and promoting germination and rooting. In practice, the undiluted solution or diluted solution can be used depending on the actual condition of the roots and the expected effect. For example, for roots with low activity or requiring strong treatment, the undiluted solution can be used directly for soaking; while for roots with good activity or requiring mild treatment, the germination and rooting solution can be diluted with water at a certain ratio (such as 1:10 or 1:20) before use, which also helps to reduce treatment costs. The choice of soaking time is also flexible. For example, when the ambient temperature is high or the roots have a strong absorption capacity, soaking for 1 hour can be chosen to shorten the treatment cycle; while when the ambient temperature is low or the roots are large and have a weak absorption capacity, soaking for 2 hours can be chosen to ensure full absorption of the active ingredients. Soaking methods can include completely immersing the roots in the liquid, or using partial immersion, spraying, etc.

[0070] After soaking, one application method is to directly transplant the roots of *Peucedanum praeruptorum* into prepared soil or other planting media. This method simplifies the operation process, reduces intermediate steps, and is particularly suitable for scenarios requiring rapid or large-scale planting, effectively improving planting efficiency.

[0071] As another application method, this method can also be used to germinate the soaked Peucedanum praeruptorum roots in the dark at 15-25℃ for 2-3 days before planting. This step aims to provide a more optimized germination environment for the Peucedanum praeruptorum roots, thereby further improving their germination and rooting success rate and consistency. The "15-25℃" temperature range is a suitable temperature determined based on the physiological characteristics of the Peucedanum praeruptorum roots. For example, at 15℃, the physiological activity of the roots can be slowly activated, suitable for situations requiring a longer germination time; while at 25℃, the physiological activity of the roots can be accelerated, suitable for situations requiring faster germination. This temperature can be maintained using temperature control equipment (such as a constant temperature chamber) or by selecting a suitable natural environment. Simultaneously, the "dark-shielding" treatment is to avoid interference from light on the physiological activity of the roots in the early stages of germination, reduce water evaporation, simulate an underground environment, and thus promote better root growth. Dark-shielding can be achieved by placing the roots in a dark container, covering them with opaque material, or placing them in a dark room. The "2-3 day germination period" setting is as follows: 2 days is suitable for seed roots with good activity and rapid germination; 3 days is suitable for seed roots with moderate activity and slow germination, ensuring that the seed roots have enough time for physiological preparation and laying a solid foundation for subsequent planting. During this germination period, suitable humidity is usually maintained, for example, through regular spraying or placing a moist medium.

[0072] Through the above technical solution, this application provides a clear and flexible method for applying the root-promoting solution for *Peucedanum praeruptorum* seeds, effectively solving the problem of inconsistent germination effects or low efficiency caused by unclear application methods in existing technologies. Specifically, by soaking the *Peucedanum praeruptorum* seeds in the undiluted or diluted root-promoting solution for 1-2 hours, it is ensured that the seeds can fully absorb the effective components in the rooting solution, laying the foundation for subsequent germination and rooting. Subsequently, two optional planting paths are provided: one is direct planting, which is simple and time-saving, suitable for planting scenarios with high efficiency requirements; the other is germination at 15-25℃ in the dark for 2-3 days before planting. This path, through precise control of temperature and light conditions, provides the optimal germination environment for the seeds, significantly improving the success rate and consistency of germination and rooting, especially suitable for scenarios with higher requirements for planting success rate and seedling quality. These two application methods complement each other, maximizing the effectiveness of the root-promoting solution, thereby ensuring the healthy growth and high yield of *Peucedanum praeruptorum* seeds.

[0073] The following example will provide a more detailed explanation of the above technical solution: In an agricultural production base, in order to improve the germination rate and rooting effect of Peucedanum praeruptorum seeds, especially in areas with poor soil conditions or high risk of drought, technicians decided to adopt a new method for preparing a germination and rooting solution.

[0074] First, the raw materials are processed. Technicians collect vigorous young willow branches in the spring. The bark of these branches is carefully peeled off and then thoroughly washed with clean water to remove surface dust and impurities. The washed bark is then cut into small pieces about 1.5 cm in length to increase the surface area during the subsequent fermentation process.

[0075] Next, fermentation was carried out. Small pieces of fresh willow bark, cut into sections, were placed in a clean fermentation tank, and an appropriate amount of water was added to achieve the preset solid-liquid ratio. Then, technicians added a compound microbial agent containing Aspergillus niger, Trichoderma, and Lactobacillus plantarum, at an inoculum size of 0.2% of the total mass of the mixture. After adding the agent, food-grade citric acid was added to precisely adjust the pH of the mixture to 5.5. The temperature inside the fermentation tank was strictly controlled at 30°C. Under these conditions, the microbial agent began to decompose plant cell walls, releasing naturally occurring growth regulators from the willow bark and synthesizing new bioactive substances. The fermentation process lasted 72 hours, with regular stirring to ensure uniform fermentation.

[0076] In another embodiment, the fermentation process can begin with enzymatic hydrolysis. Technicians first add a compound enzyme preparation of cellulase and pectinase to a mixture of willow twig bark fragments and water, with a total addition amount of 0.1% of the total mass of the mixture. The enzymatic hydrolysis conditions are set at 45°C for 3 hours. After enzymatic hydrolysis, the aforementioned compound microbial agent is added, and the pH is adjusted to 5.5, followed by microbial fermentation at 30°C. This strategy of enzymatic hydrolysis followed by fermentation can more thoroughly decompose plant cell walls and improve the release efficiency of active ingredients.

[0077] Finally, post-processing is performed. After fermentation, a fermentation broth rich in bioactive substances is obtained. This broth is then subjected to solid-liquid separation using a centrifuge to remove solid residues, thus obtaining a preliminary germination and rooting solution. To facilitate storage and transportation, and to further improve its active concentration, technicians performed vacuum concentration on the separated germination and rooting solution to ensure that the content of its effective components reaches the predetermined standard. Simultaneously, to extend the product's shelf life, an appropriate amount of food-grade preservative, such as potassium sorbate, is added to the concentrated liquid.

[0078] like Figure 2 As shown, after using the germination solution of this embodiment to germinate the roots of Peucedanum praeruptorum, the number of germinated roots reached nearly 10.

[0079] To scientifically verify the effectiveness of the technical solution in this embodiment, the applicant designed the following three sets of comparative experiments. All experiments were conducted within the same time period and from the same batch of Peucedanum praeruptorum roots to control variables to the greatest extent possible.

[0080] I. Core Comparative Experimental Data (Indoor Germination and Rooting Experiment) 1. Experimental Design: Test material: Healthy, uniformly sized roots of Peucedanum praeruptorum (diameter 1.0±0.2cm).

[0081] Experimental group: a. CK (blank control): Soaked in water.

[0082] b. CA (Chemical Control): Immersion in 50 mg / L indolebutyric acid (IBA) solution (commercially available standard procedure).

[0083] c. TW (Traditional Control): Soak fresh willow bark in the supernatant after soaking at room temperature for 48 hours at a ratio of 1:10 (w / v).

[0084] d. TF-A (this invention): Fermentation stock solution prepared according to scheme A (microbial fermentation), diluted 5 times and then soaked.

[0085] e. TF-B (this invention): The fermentation stock solution prepared according to scheme B (enzymatic hydrolysis-fermentation combination) is diluted 5 times and then soaked.

[0086] Treatment method: Soak the seed roots for 2 hours, then remove them and place them in a petri dish lined with moistened filter paper. Incubate at a constant temperature of 25℃ in the dark. Each treatment was repeated 3 times, with 50 seed root segments per replicate.

[0087] Observation period: days 5, 10, and 15.

[0088] 2. Observation Results Data Table: 3. Conclusion: The TF-A and TF-B treatments of this invention are significantly superior to the blank control (CK), chemical hormone control (CA), and traditional willow twig sap control (TW) in all key indicators. Among them, TF-B (enzymatic hydrolysis-fermentation combined process) has the most outstanding effect, indicating that this process can release active substances more efficiently.

[0089] The above description describes specific embodiments of the present invention, but the scope of protection of the present invention is not limited thereto. Any person skilled in the art can easily conceive of various equivalent modifications or substitutions within the technical scope disclosed in the present invention, and these modifications or substitutions should all be covered within the scope of protection of the present invention. Therefore, the scope of protection of the present invention should be determined by the scope of the claims.

Claims

1. A method for preparing a root-promoting and germination-enhancing solution for Peucedanum praeruptorum seeds, characterized in that, Includes the following steps: S1. Raw material processing: Collect tender willow branches, peel off their fresh bark, wash them, and cut them into small sections; S2. Fermentation treatment: Mix the small pieces of fresh bark from the tender willow branches with water, add a fermentation aid, adjust the pH to acidic, and ferment at 25-35℃ to obtain a fermentation liquid; the fermentation aid is a microbial agent and / or enzyme preparation that can decompose plant cell walls. S3. Post-processing: The fermentation broth is subjected to solid-liquid separation to obtain the sprouting and rooting solution.

2. The preparation method according to claim 1, characterized in that, In step S2, the pH value is adjusted to 4-6.

3. The preparation method according to claim 1 or 2, characterized in that, In step S2, the fermentation temperature is 28-32°C.

4. The preparation method according to claim 1, characterized in that, In step S2, the microbial agent includes at least one of Aspergillus niger, Trichoderma, and Lactobacillus plantarum.

5. The preparation method according to claim 4, characterized in that, The microbial agent is a compound agent containing Aspergillus niger, Trichoderma and Lactobacillus plantarum, and its inoculum amount is 0.1%-0.5% of the total mass of the mixed system.

6. The preparation method according to claim 1, characterized in that, In step S2, the enzyme preparation includes cellulase and pectinase; the fermentation process specifically includes: first adding the enzyme preparation for enzymatic hydrolysis, and then adding the microbial agent for fermentation.

7. The preparation method according to claim 6, characterized in that, The enzymatic hydrolysis conditions are: temperature 40-50℃, time 2-4 hours; the total amount of enzyme preparation added is 0.05%-0.2% of the total mass of the mixed system.

8. The preparation method according to claim 1, characterized in that, In step S3, the solid-liquid separation is followed by a concentration and / or the addition of a preservative.

9. The use of a germination and rooting solution prepared by the method of any one of claims 1-8 in promoting the germination and / or rooting of Peucedanum praeruptorum seeds.

10. The application according to claim 9, characterized in that, The application method includes: soaking the roots of Peucedanum praeruptorum in the undiluted or diluted rooting solution for 1-2 hours before planting; or, germinating the soaked roots in the dark at 15-25℃ for 2-3 days before planting.

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