A planting method for controlling and preventing fusarium wilt by using stephania sinica and woody plant undergrowth rotation

Abstract

The application provides a planting method for preventing and controlling fusarium wilt by under-forest rotation of Stephania tetrandra S. Moore and woody plants. The under-forest rotation planting system of Stephania tetrandra S. Moore realizes all-round improvement from soil improvement to biological defense through the synergistic application of various technologies, and ultimately realizes ecological and sustainable prevention and control of fusarium wilt in disease prevention and control and production safety. The disease occurrence rate is fundamentally curbed, and the stable production and high yield of Stephania tetrandra S. Moore are ensured. A comprehensive technical paradigm that can be replicated and popularized is provided for the green and high-quality development of the entire traditional Chinese medicinal material industry, and the ecological benefit and economic benefit are integrated.

Description

Technical Field

[0001] This invention belongs to the technical field of Chinese medicinal herb cultivation, specifically relating to a cultivation method for controlling Fusarium wilt disease through understory rotation of Stephania tetrandra and other woody plants. Background Technology

[0002] As an important traditional Chinese medicinal herb, Stephania tetrandra faces a severe challenge in its large-scale production from soil-borne diseases, particularly Fusarium wilt caused by Fusarium. Currently, the control of this disease relies primarily on chemical pesticides. However, this traditional method has significant limitations: First, long-term, singular use of chemical pesticides easily induces drug resistance in pathogens, leading to a gradual decline in control effectiveness. Second, while killing pathogens, chemical pesticides indiscriminately inhibit the activity of beneficial microorganisms in the soil, disrupting the soil's microecological balance, causing loss of soil biodiversity, and deteriorating its physical and chemical properties, creating a vicious cycle of "the more you try to control it, the more disease it gets." Finally, pesticide residues directly affect the safety of using medicinal herbs, failing to meet the requirements of green agriculture and high-quality medicinal herb production. Therefore, relying solely on chemical control is not only unsustainable but also poses a potential threat to the authentic quality of the medicinal herbs and the ecological environment of the production area.

[0003] On the other hand, in the pursuit of high-yield and high-efficiency intensive planting models, the soil is often over-exploited, and its health is neglected. Common problems in southern production areas, such as soil acidification and compaction, directly affect the development and function of the root system of Stephania tetrandra. A healthy root system is the foundation for disease resistance and efficient synthesis of medicinal components. However, in poor soil environments, plants grow weakly, have poor resistance to adverse conditions, and are more susceptible to disease. At the same time, the accumulation of secondary metabolites (such as tetrandrine and other core medicinal components) is also inhibited, leading to fluctuations or even a decline in the intrinsic quality of the medicinal material, making it difficult to stably guarantee the market reputation and value of "authentic medicinal materials".

[0004] Furthermore, the traditional monoculture continuous planting model exacerbates the accumulation and spread of soil-borne diseases, leading to the singular output of land resources and the degradation of ecosystem services. How to achieve sustainable disease control, stable improvement in medicinal herb quality, and maximization of land productivity without sacrificing the ecological environment is a core technological bottleneck that the current Stephania tetrandra industry must overcome for transformation and upgrading. Simply introducing a single biological agent or plant amendment often yields limited and unstable results because it fails to systematically reconstruct a rhizosphere micro-ecosystem that is conducive to crop health and unfavorable to pathogen growth.

[0005] Therefore, the industry urgently needs a comprehensive technical solution that can synergistically improve the physical, chemical, and biological properties of soil, and integrate the principles of niche management and biological synergistic defense. This solution requires a multi-dimensional approach, simultaneously addressing factors such as creating a soil environment unfavorable to pathogen survival, directly inhibiting pathogens and pests, and strengthening crop immunity and nutrient absorption, to construct a stable, self-sustaining healthy interaction system between soil, microorganisms, and plants. This is not only key to overcoming Fusarium wilt, but also an inevitable path to transform the cultivation of Stephania tetrandra into a sustainable development model that is environmentally friendly, resource-saving, and prioritizes quality. It has significant practical implications for ensuring the safety of medicinal herb sources and enhancing the overall competitiveness of the industry. Summary of the Invention

[0006] This invention discloses a planting method for controlling Fusarium wilt disease through understory rotation of Stephania tetrandra and woody plants, to solve any of the above-mentioned or potential problems in the prior art. To solve the above-mentioned technical problems, the specific steps of this application are as follows: (1) Soil conditioning: 1-2 months before planting Stephania tetrandra, the soil is treated as follows: evenly spread 300-400 parts of biochar, 0-20 parts of gypsum, 0.5-1 parts of organic humic acid, 0-200 parts of oyster shell powder, 25-35 parts of neem powder and 0.5-1 parts of vetiver gum per mu; after spreading, mix the materials with the soil thoroughly by deep plowing or rotary tillage, with a plowing depth of 20-25 cm; then, apply 300-500 parts of organic fertilizer per mu. Preferably, the organic matter content of the organic fertilizer is not less than 45%; The preparation process of oyster shell powder is as follows: after cleaning and drying oyster shells, crush them into fragments of 1-3 cm; place the fragments in an oxidizing atmosphere kiln and calcine them at a temperature of 850-950℃ for 1.5-3 hours; after the calcined product cools naturally, age it in a dry and ventilated environment for 7-14 days; finely grind the aged material and pass it through an 80-100 mesh sieve to obtain the oyster shell powder.

[0007] The soil in Jiangxi Province is mostly acidic, and the soil fertility is generally low.

[0008] If the soil pH is <5.5 (strongly acidic): use 150-200 kg / mu of oyster shell powder.

[0009] If the soil pH is between 5.5 and 6.0, 100 kg / mu of oyster shell powder can be used.

[0010] If the soil pH is between 6.0 and 7.0: there is no need to use oyster shell powder for pH adjustment, but a small amount (e.g., 50 kg / acre) can still be applied as a calcium fertilizer supplement.

[0011] If the soil pH is >7.0 (alkaline): the use of oyster shell powder should be completely stopped, and acidity regulators such as sulfur powder should be considered.

[0012] (2) Application of fungal agents: The application of fungal agents includes the use of Trichoderma harzianum agent and Gyrodactylis mosieurinum agent, wherein Trichoderma harzianum agent is a solid powder; Gyrodactylis mosieurinum agent is a liquid fungal agent; the number of effective live spores is not less than 1×10 8 CFU / g, Moses Gastrodia elata fungicide contains no less than 200 infectious vegetative cells per gram; The application of microbial agents is divided into three stages: The first stage is root dipping during the seedling stage: Take 0.5 parts of Trichoderma harzianum agent and 1.0 part of Gastrodia elata agent, add 0.05 parts of Vilan gum and 0.5 parts of brown sugar, and mix with 20-30 parts of water at a temperature of 20-28℃ to make a root dipping solution; immerse the roots of Stephania tetrandra seedlings in the root dipping solution for 15-20 minutes, and then transplant them. The second stage is the application of the fertilizer in the planting holes: Mix the remaining Trichoderma harzianum and Trichoderma mosesi with organic fertilizer at a weight ratio of 1:15 to 1:20 to make bio-fertilizer; apply 0.03-0.05 parts of the bio-fertilizer in each planting hole, cover with a thin layer of soil, and then plant the seedlings that have been treated with root dipping. The third stage is topdressing during the growing season: one month after transplanting and the seedlings have recovered, dilute 0.2-0.3 parts of Trichoderma harzianum agent with water 300-400 times to make a fungal suspension, and drench the roots with 200-300 ml per plant. (3) Understory rotation: Select at least one of the following woody plants for rotation: Camellia oleifera, walnut, Eucommia ulmoides, Magnolia officinalis, or Cunninghamia lanceolata. Plant the woody plants at a density of 3 meters between plants and 4 meters between rows. Make ridges between the rows of the woody plants and plant 1-2 rows of Stephania tetrandra, with a spacing of not less than 1 meter between the Stephania tetrandra rows and the woody plant rows. The row spacing of Stephania tetrandra is 60-80 cm and the plant spacing is 30-40 cm. These woody plants are selected for understory rotation because they complement Stephania tetrandra in terms of ecological niche. The canopy of the woody plants can regulate the microenvironment, reduce the spread of soil diseases, and improve land use efficiency and economic benefits, which is in line with the principles of ecological agriculture.

[0013] The advantages and beneficial effects of this invention are as follows: The understory rotation planting system of Stephania tetrandra in this invention achieves a comprehensive improvement from soil improvement to biological defense through the synergistic application of multiple technologies, ultimately bringing significant beneficial effects in many aspects, including disease control, medicinal quality, yield, ecological benefits and economic returns.

[0014] In the initial soil improvement phase, vegan gum and calcined oyster shell powder together laid the foundation for a soil environment that promotes growth and suppresses disease. As a highly efficient biopolymer, vegan gum's core function lies in its strong binding effect, firmly binding additives such as biochar to soil particles to form durable and porous water-stable aggregates. This not only directly optimizes the aeration and water retention performance of the root zone, but more importantly, it creates a structurally stable "micro-habitat" for subsequently introduced beneficial microorganisms, free from drastic environmental fluctuations, ensuring the sustainability of biological control effects. Simultaneously, calcined oyster shell powder precisely regulates the soil's chemical environment. Addressing the common problem of severe soil acidification in hilly areas of southern China, it slowly releases alkaline substances, effectively raising the soil pH from an extremely unfavorable acidic range to the ideal range of slightly acidic to neutral.

[0015] Building upon a solid foundation of excellent soil physicochemical properties, neem powder and functional microorganisms form a synergistic defensive biological mechanism. As a plant-derived biopesticide, neem powder continuously releases azadirachtin and other active substances that effectively inhibit the germination of Fusarium spores and repel and kill root-knot nematodes and other underground pests. This significantly reduces the initial pathogen population in the soil during the early stages of crop growth and minimizes root wounds caused by pests. This establishes an early chemical protective barrier for Stephania tetrandra seedlings.

[0016] In subsequent root dipping during the seedling stage, the targeted inoculation of *Trichoderma harzianum* and *Gloydium moses* rapidly colonized and established dominant populations. *Trichoderma harzianum* eradicated Fusarium around the rhizosphere through multiple mechanisms, including rapid competition for nutrients and space, entanglement with and parasitization of pathogenic fungal hyphae, and antibiotic secretion. Meanwhile, the hyphae of *Gloydium moses* formed a symbiotic relationship with the roots of *Stephania tetrandra*, significantly expanding the plant's absorption network for nutrients such as phosphorus and zinc, as well as water, fundamentally improving plant health physiologically. Simultaneously, this symbiotic relationship systematically induced broad-spectrum resistance in the plant, placing it in a highly immune state. Under favorable conditions created by *Neemum neem*, they jointly constructed a living, self-sustaining, and proliferating rhizosphere biological defense barrier. Multi-stage targeted inoculation aims to systematically establish and maintain a beneficial rhizosphere microbial community, inhibiting soil-borne pathogens such as Fusarium. *Trichoderma harzianum* directly controlled wilt disease through competition and antagonism; *Gloydium moses*, as an arbuscular mycorrhizal fungus, promoted plant nutrient absorption and disease resistance. Root dipping treatment ensures that the inoculant is directly colonized on the root system; planting hole application provides a continuous source of inoculants; and top dressing during the growing season strengthens the dominance of rhizosphere microorganisms, thereby reducing the risk of disease and reducing reliance on chemical pesticides.

[0017] Ultimately, in terms of disease control and production safety, this system achieves ecological and sustainable control of Fusarium wilt, fundamentally curbing the disease's incidence and ensuring stable production of Stephania tetrandra while avoiding the risks of chemical pesticide use and residues, thus guaranteeing the safety of the medicinal material at its source. Secondly, regarding the quality and authenticity of the medicinal material, Stephania tetrandra grown in optimized and healthy soil and microbial environments exhibits more vigorous physiological metabolism. Studies have confirmed that under favorable mycorrhizal symbiosis and balanced nutritional conditions, the synthesis and accumulation of medicinal active ingredients in its roots (such as tetrandrine and tetrandrine) are more complete, significantly enhancing the intrinsic quality of the medicinal material and the authentic characteristics of "Jiangxi Stephania tetrandra." Finally, in terms of ecological and industrial sustainability, this method, with the cultivation of healthy soil and biodiversity as its core, combines land use with soil conservation, transforming the soil from a consumable resource into a self-sustaining living asset. By combining the economic output of understory woody plants (such as camellia oleifera), a composite management model of "using short-term gains to support long-term development and ecological cycle" has been formed. This not only brings additional long-term economic benefits to growers, but also provides a replicable and scalable comprehensive technical paradigm for the green and high-quality development of the entire Chinese medicinal materials industry. Attached Figure Description

[0018] Figure 1 This is a diagram of the early seedling cultivation of Stephania tetrandra.

[0019] Figure 2 This is a picture showing the results of cultivating Stephania tetrandra alone.

[0020] Figure 3 Seedling cultivation diagram for intercropping Stephania tetrandra in Camellia oleifera forest.

[0021] Figure 4 A picture showing the later results of intercropping Stephania tetrandra in Camellia oleifera forest. Detailed Implementation

[0022] The present invention will be further described in detail below with reference to the embodiments.

[0023] The organic fertilizer used in the following implementation methods was purchased from Hebei Sanpin Fertilizer Co., Ltd., and its content standard complies with NY884-2012, with an organic matter content of not less than 45%.

[0024] Trichoderma harzianum fungicide is a solid powder with an effective viable spore count of not less than 1×10⁻⁶. 8 CFU / g, purchased from Shandong Changtai Biotechnology Co., Ltd.; Moses Globulus fungicide, containing no less than 200 infective vegetative cells per gram, purchased from Nanjing Cuijingyuan Biotechnology Co., Ltd.; Neem powder, extracted from the seeds and leaves of Neem tree (a plant in the genus Neem of the family Meliaceae), is a brownish-yellow powder with a fineness of 100 mesh, purchased from Lanzhou Waterles Biotechnology Co., Ltd.

[0025] The following implementation methods were conducted at a Stephania tetrandra planting base in Jiangxi Province. The test soil was red soil with an initial pH of 5.8, classifying it as acidic. The previous crop was Stephania tetrandra, which had a history of wilt disease. The experiment began in the spring of 2023. In the following implementation methods, two-year-old Camellia oleifera was selected as a woody plant for crop rotation. The implementation field for each implementation method was 1 mu (approximately 0.16 acres).

[0026] The preparation process of oyster shell powder is as follows: take oyster shells, clean and dry them, and break them into fragments of about 2 cm; place the fragments in an oxidizing atmosphere rotary kiln and calcine them at 900℃ for 2 hours; after the product cools naturally, it is aged in a rainproof shed for 10 days; finally, it is ground and passed through a 90-mesh sieve to obtain the desired oyster shell powder. Example 1

[0027] Forty-five days before planting, evenly spread 350 kg of biochar, 0.8 kg of organic humic acid, 100 kg of oyster shell powder, 30 kg of neem powder, and 0.8 kg of vetiver per acre, and mix thoroughly by rotary tilling to a depth of 22 cm. Subsequently, apply 400 kg of commercial organic fertilizer per acre.

[0028] Seedling root dipping: Take 0.5 kg of Trichoderma harzianum agent and 1.0 kg of Gastrodia moses agent, add 0.05 kg of vegan gum and 0.5 kg of brown sugar, and mix with 25 liters of 25℃ water to make a bacterial solution. Soak the seedling roots for 18 minutes.

[0029] Application in planting holes: Mix the remaining inoculant with organic fertilizer at a ratio of 1:18 to obtain bio-fertilizer, and apply 40 grams of bio-fertilizer to each planting hole.

[0030] Topdressing during the growing season: 30 days after transplanting Stephania tetrandra, mix 0.25 kg of Trichoderma harzianum powder with 100 liters of water and drench the roots of each plant with 250 ml of the solution.

[0031] The understory configuration is as follows: Camellia oleifera plants are spaced 3 meters apart and 4 meters apart. Two rows of Stephania tetrandra are planted on raised beds between the rows. The row spacing of Stephania tetrandra is 70 centimeters and the plant spacing is 35 centimeters. The edge of the raised bed is 1.2 meters away from the Camellia oleifera row. Example 2

[0032] Two months before planting, apply 300 kg of biochar, 0.5 kg of organic humic acid, 180 kg of oyster shell powder, 25 kg of neem powder, and 0.5 kg of venetian gum per mu. After rotary tilling to a depth of 20 cm, apply 300 kg of organic fertilizer.

[0033] Root dipping with fungicide: Take 0.5 kg of Trichoderma harzianum fungicide and 1.0 kg of Gastrodia moses fungicide, add 0.05 kg of Vilan gum and 0.5 kg of brown sugar, mix with 20 liters of 22°C water, and soak the seedling roots for 15 minutes.

[0034] Application in planting holes: Mix the remaining inoculant with organic fertilizer at a ratio of 1:15 to obtain bio-fertilizer, and apply 30 grams per hole.

[0035] Topdressing during the growing season: One month after transplanting, dilute 0.2 kg of Trichoderma harzianum powder with 60 liters of water and drench the roots of each plant with 200 ml of the solution.

[0036] The understory configuration is as follows: Camellia oleifera plants are spaced 3 meters apart and 4 meters apart, with one row of Stephania tetrandra planted between the rows. Stephania tetrandra plants are spaced 80 centimeters apart in rows and 40 centimeters apart in plants, and are 1 meter away from the Camellia oleifera row. Example 3

[0037] One month before planting, apply 400 kg of biochar, 15 kg of gypsum, 1 kg of organic humic acid, 50 kg of oyster shell powder, 35 kg of neem powder, and 1 kg of vetiver per mu. After rotary tilling to a depth of 25 cm, apply 500 kg of organic fertilizer.

[0038] Root dipping with fungicide: Take 0.5 kg of Trichoderma harzianum fungicide and 1.0 kg of Gastrodia moses fungicide, add 0.05 kg of Vilan gum and 0.5 kg of brown sugar, mix with 30 liters of 22°C water, and soak the seedling roots for 15 minutes.

[0039] Application in planting holes: Mix the remaining inoculant with organic fertilizer at a ratio of 1:20 to obtain bio-fertilizer, and apply 50 grams per hole.

[0040] Topdressing during the growing season: One month after transplanting, dilute 0.3 kg of Trichoderma harzianum powder with 120 liters of water and drench the roots of each plant with 300 ml of the solution.

[0041] The understory configuration is as follows: Camellia oleifera plants are spaced 3 meters apart and 4 meters apart, with two rows of Stephania tetrandra planted between the rows. Stephania tetrandra plants are spaced 60 centimeters apart in rows and 30 centimeters apart in plants, and are 1.5 meters away from the Camellia oleifera rows.

[0042] Comparative Example 1 Forty-five days before planting, evenly spread 350 kg of biochar, 0.8 kg of organic humic acid, 100 kg of oyster shell powder, 30 kg of neem powder, and 0.8 kg of vetiver per acre, and mix thoroughly by rotary tilling to a depth of 22 cm. Subsequently, apply 400 kg of commercial organic fertilizer per acre.

[0043] Seedling root dipping: Take 0.5 kg of Trichoderma harzianum agent and 1.0 kg of Gastrodia moses agent, add 0.05 kg of vegan gum and 0.5 kg of brown sugar, and mix with 25 liters of 25℃ water to make a bacterial solution. Soak the seedling roots for 18 minutes.

[0044] Application in planting holes: Mix the remaining inoculant with organic fertilizer at a ratio of 1:18 to obtain bio-fertilizer, and apply 40 grams of bio-fertilizer to each planting hole.

[0045] Topdressing during the growing season: 30 days after transplanting Stephania tetrandra, mix 0.25 kg of Trichoderma harzianum powder with 100 liters of water and drench the roots of each plant with 250 ml of the solution.

[0046] The planting pattern is monoculture of Stephania tetrandra, which means that the plant is planted in open fields without ridges, with a row spacing of 60 cm and a plant spacing of 30 cm.

[0047] Comparative Example 2 Forty-five days before planting, evenly spread 350 kg of biochar, 0.8 kg of organic humic acid, 100 kg of oyster shell powder, 30 kg of neem powder, and 0.8 kg of vetiver per acre, and mix thoroughly by rotary tilling to a depth of 22 cm. Subsequently, apply 400 kg of commercial organic fertilizer per acre.

[0048] The understory configuration is as follows: Camellia oleifera plants are spaced 3 meters apart and 4 meters apart. Two rows of Stephania tetrandra are planted in ridges between the rows. One month after planting Stephania tetrandra, 40 grams of organic fertilizer are applied as top dressing. The spacing between Stephania tetrandra rows is 70 centimeters and the spacing between plants is 35 centimeters. The edge of the ridge is 1.2 meters away from the Camellia oleifera row.

[0049] Comparative Example 3 Forty-five days before planting, evenly spread 350 kg of biochar, 0.8 kg of organic humic acid, 100 kg of oyster shell powder, 30 kg of neem powder, and 0.8 kg of vetiver per acre, and mix thoroughly by rotary tilling to a depth of 22 cm. Subsequently, apply 400 kg of commercial organic fertilizer per acre.

[0050] Take 0.5 kg of Trichoderma harzianum agent and 1.0 kg of Gastrodia elata agent, add 0.05 kg of velan gum and 0.5 kg of brown sugar, mix with 25 liters of 25℃ water to make a bacterial solution, and mix with organic fertilizer at a ratio of 1:18 to obtain a bio-fertilizer. When preparing the land, spread it evenly on the soil surface and lightly rake it.

[0051] The understory configuration is as follows: Camellia oleifera plants are spaced 3 meters apart and 4 meters apart. Two rows of Stephania tetrandra are planted on raised beds between the rows. The row spacing of Stephania tetrandra is 70 centimeters and the plant spacing is 35 centimeters. The edge of the raised bed is 1.2 meters away from the Camellia oleifera row.

[0052] Comparative Example 4 The difference between this comparative example and Example 1 is that oyster shell powder is replaced with quicklime in this comparative example; otherwise, it is the same as Example 1.

[0053] Comparative Example 5 The difference between this comparative example and Example 1 is that in this comparative example, neem powder is replaced with 3% phoxim granules as an insecticide; otherwise, it is the same as in Example 1.

[0054] Comparative Example 6 The difference between this comparative example and Example 1 is that in this comparative example, vegan gum is replaced with xanthan gum; otherwise, it is the same as in Example 1.

[0055] Comparative Example 7 The difference between this comparative example and Example 1 is that in this comparative example, all Trichoderma harzianum agents were replaced with an equal number of effective viable spores (2 × 10⁻⁶). 8 Bacillus subtilis solid inoculum (CFU / g); the rest is the same as in Example 1.

[0056] Experiment 1: Disease survey of Fusarium wilt in Stephania tetrandra All treatments of Stephania tetrandra were transplanted around mid-April 2023. After transplanting, except for the specific measures prescribed for each treatment, other field management practices (such as irrigation, weeding, trellising, etc.) were kept consistent and followed local conventional production and management standards for Stephania tetrandra. Throughout the entire growth period, except for Comparative Example 5, no chemical pesticides or fungicides were used in any of the treatments.

[0057] The "five-point sampling method" was used in each group, with 20 plants randomly and continuously investigated at each sampling point, for a total of 100 plants investigated.

[0058] Based on the symptoms of the above-ground parts of the plant, wilt disease is classified into 5 levels (0-4): Grade 0: The entire plant is healthy and shows no signs of disease.

[0059] Grade 1: A few leaves at the base of the plant show signs of chlorosis or yellowing, or one side of the branches shows slight wilting.

[0060] Level 2: The lower and middle leaves of the plant are generally yellow, and less than 1 / 3 of the plant is wilting.

[0061] Level 3: Most of the plant's leaves are yellowed and wilted, and 1 / 3 to 2 / 3 of the plant wilts or dies.

[0062] Level 4: The entire plant is severely wilted and dead.

[0063] Disease severity index calculation: The disease severity index for each community is calculated using the following formula: Disease Index (DI, %) = [ [(Number of tested plants × highest representative value)] × 100 In this calculation, the representative value for each disease level is its level number (0, 1, 2, 3, 4). The total number of plants surveyed is 100, and the highest disease level representative value is 4.

[0064] Control efficacy (CE) calculation: Using CP1 (monoculture control) as the baseline control, the control efficacy of other intercropping treatments in the forest was calculated. Simultaneously, CP2 (no microorganisms in the forest understory control) was used as a secondary control to evaluate the control efficacy of microbial inoculants. The calculation formula is as follows: Control efficacy (CE, %) = Disease index in control area - Disease index in treatment area / Disease index in control area × 100 Control efficacy (CE, %) = [(Disease index in control area - Disease index in treatment area) / Disease index in control area] × 100 The results are shown in Table 1 below: Table 1. Disease index and control effect of Stephania tetrandra wilt in each treatment group Example 1 1.75 96.22 94.40 Example 2 2.50 94.60 92.00 Example 3 4.00 91.35 87.20 Comparative Example 1 46.25 - - Comparative Example 2 31.25 32.43 - Comparative Example 3 13.75 70.27 56.00 Comparative Example 4 12.50 72.97 60.00 Comparative Example 5 9.50 79.46 69.60 Comparative Example 6 5.00 89.20 84.00 Comparative Example 7 6.00 87.03 80.80 Experiment 2: Yield Measurement Harvesting will take place in late October 2025, after the above-ground parts of Stephania tetrandra have naturally withered.

[0065] Yield measurement method: Select a 20m² quadrat in the central area of ​​each group, dig out the tuberous roots of all Stephania tetrandra plants in the quadrat completely, remove the soil and fibrous roots, wash with clean water and drain the surface water, drain the surface water in a cool and ventilated place, and then use an electronic scale to weigh the fresh weight (kg) of all tuberous roots in the quadrat.

[0066] Yield conversion: Based on the fresh weight of the sample plots, the fresh weight yield per mu (667m²) was calculated (kg / mu). Since Stephania tetrandra has a long growth cycle, this study measured the yield two and a half years after planting, which better reflects the differences between treatments.

[0067] Yield per mu (kg / mu) = [fresh weight of sample plot (kg) / 20m] 2 】×667m 2 / mu The results are shown in Table 2 below: Table 2 Fresh weight yield of Stephania tetrandra tubers in each treatment group Example 1 45.52 1518.18 Example 2 43.88 1463.49 Example 3 42.50 1417.48 Comparative Example 1 26.39 880.15 Comparative Example 2 29.80 993.89 Comparative Example 3 29.09 970.22 Comparative Example 4 32.50 1083.98 Comparative Example 5 24.59 820.12 Comparative Example 6 33.74 1125.30 Comparative Example 7 35.09 1170.30 Experiment 3: Detection of main pharmaceutical active ingredients: Sample preparation: 1 kg was randomly selected from the tuberous root samples of each treatment group, washed and cut into thin slices, dried in an oven at 60℃ to constant weight, pulverized and passed through an 80-mesh sieve to obtain powdered Stephania tetrandra medicinal material, which was sealed and stored in a desiccator for later use.

[0068] Detected components: Tetrandrine, Fangchinoline, and total alkaloids.

[0069] Detection method: Quantitative analysis was performed using high performance liquid chromatography (HPLC) in accordance with the Pharmacopoeia of the People's Republic of China (2020 edition).

[0070] Chromatographic conditions: Column: Agilent ZORBAX SB-C18 (4.6 mm × 250 mm, 5 μm) or a C18 reversed-phase column of equivalent performance. Mobile phase: Methanol-0.1% triethylamine aqueous solution (80:20, v / v), adjusted to pH 7.5 with phosphoric acid. Detection wavelength: 282 nm. Flow rate: 1.0 mL / min. Column temperature: 30 °C. Injection volume: 10 μL.

[0071] Preparation of reference solutions: Accurately weigh appropriate amounts of tetrandrine reference standard and tetrandrine reference standard (purity ≥98%, purchased from the China National Institutes for Food and Drug Control), dissolve and dilute with methanol to prepare a series of mixed standard solutions containing tetrandrine and tetrandrine concentrations of 5, 10, 20, 40, and 80 μg / mL, respectively.

[0072] Preparation of test solution: Accurately weigh approximately 0.5 g of herbal powder from each treatment group and place it in a stoppered conical flask. Accurately add 50 mL of mobile phase, seal tightly, weigh, and sonicate (250 W, 40 kHz) for 45 minutes. Remove and cool, shake well, and let stand or centrifuge at 5000 rpm for 5 minutes. Weigh again, replenish the lost weight with mobile phase, shake well, and filter through a 0.45 μm microporous membrane. Take the supernatant filtrate as the test solution. Prepare three replicates for each sample.

[0073] Content Calculation: A standard curve was plotted with peak area on the ordinate and reference concentration on the abscissa. Based on the peak area of ​​the test solution, the concentrations of tetrandrine and tebufenozide were calculated from the standard curve and then converted to their percentage content in the medicinal material. The total alkaloid content was calculated as the sum of the contents of tetrandrine and tebufenozide.

[0074] The results are shown in Table 3 below: Table 3. Content of main active ingredients in the tuberous roots of Stephania tetrandra in each treatment group Example 1 1.58 0.92 2.50 Example 2 1.51 0.87 2.38 Example 3 1.45 0.84 2.29 Comparative Example 1 1.05 0.61 1.66 Comparative Example 2 1.18 0.65 1.83 Comparative Example 3 1.15 0.63 1.78 Comparative Example 4 1.25 0.70 1.95 Comparative Example 5 0.88 0.53 1.41 Comparative Example 6 1.32 0.75 2.07 Comparative Example 7 1.38 0.79 2.17 Data from Tables 1 to 3 show that Examples 1-3 significantly outperformed the comparative examples in disease control, yield, and medicinal quality: the wilt disease severity index of the examples (1.75%-4.00%) was much lower than that of the monoculture control (Comparative Example 1, 46.25%), and the control effect reached over 91%; the yield per mu (1417-1518 kg) and total alkaloid content (2.29%-2.50%) were also significantly higher than those of the comparative examples. This comprehensive advantage stems from the systematic synergy of the examples through "soil conditioning - targeted microbial inoculation - understory rotation": vesiniplatin and calcined oyster shell powder optimized the soil structure and regulated the pH, creating a stable habitat for beneficial microorganisms; neem powder continuously inhibited pathogens and nematodes; the staged inoculation of Trichoderma harzianum and Gastrodia moses constructed a long-term biological defense line in the rhizosphere, directly antagonizing pathogens and enhancing plant resistance and nutrient absorption; and understory rotation (Camellia oleifera) further regulated the microenvironment and blocked disease transmission. In contrast, the comparative examples lacked key steps (e.g., Comparative Example 2 did not use microbial agents, Comparative Example 3 used microbial agents only once, Comparative Example 4 used quicklime to destroy microorganisms, Comparative Example 5 used chemical pesticides to inhibit beneficial bacteria, Comparative Example 6 had poor xanthan gum aggregation effect, and Comparative Example 7 had weak Bacillus subtilis disease prevention effect), resulting in incomplete system function and limitations in disease control, yield, and quality improvement. Therefore, the core advantage of this invention lies in its multi-level, multi-target ecological integration, simultaneously achieving green disease control, yield improvement, and the guarantee of the authenticity of medicinal materials, forming a sustainable and healthy interactive system.

Claims

1. A planting method for controlling Fusarium wilt disease through understory rotation of Stephania tetrandra and woody plants, comprising soil conditioning, application of fungicides, and understory rotation planting, characterized in that, The soil conditioning process includes the following treatment of the soil 1-2 months before planting Stephania tetrandra: Apply 300-400 parts biochar, 0-20 parts gypsum, 0.5-1 part organic humic acid, 0-200 parts oyster shell powder, 25-35 parts neem powder, and 0.5-1 part vetiver evenly per acre; after application, thoroughly mix the materials with the soil through deep plowing or rotary tillage to a depth of 20-25 cm; then, apply 300-500 parts organic fertilizer per acre. The organic matter content of the organic fertilizer shall not be less than 45%.

2. The planting method according to claim 1, characterized in that, The preparation process of the oyster shell powder is as follows: after cleaning and drying the oyster shells, crush them into fragments of 1-3 cm; place the fragments in an oxidizing atmosphere kiln and calcine them at a temperature of 850-950℃ for 1.5-3 hours; after the calcined product cools naturally, age it in a dry and ventilated environment for 7-14 days; finely grind the aged material and pass it through an 80-100 mesh sieve to obtain the oyster shell powder.

3. The planting method according to claim 1, characterized in that, The application of the fungal agents includes the use of Trichoderma harzianum agent and Gyrodactylum moses agent, wherein Trichoderma harzianum agent is a solid powder and Gyrodactylum moses agent is a liquid fungal agent.

4. The planting method according to claim 1 or 3, characterized in that, The application of the microbial agent is divided into three stages: The first stage is root dipping during the seedling stage: Take 0.5 parts of Trichoderma harzianum agent and 1.0 part of Gastrodia elata agent, add 0.05 parts of Vilan gum and 0.5 parts of brown sugar, and mix with 20-30 parts of water at a temperature of 20-28℃ to make a root dipping solution; immerse the roots of Stephania tetrandra seedlings in the root dipping solution for 15-20 minutes, and then transplant them. The second stage is the application of the fertilizer in the planting holes: Mix the remaining Trichoderma harzianum and Trichoderma mosesi with organic fertilizer at a weight ratio of 1:15 to 1:20 to make bio-fertilizer; apply 0.03-0.05 parts of the bio-fertilizer in each planting hole, cover with a thin layer of soil, and then plant the seedlings that have been treated with root dipping. The third stage is topdressing during the growing season: one month after transplanting and the seedlings have recovered, dilute 0.2-0.3 parts of Trichoderma harzianum agent with water 300-400 times to make a fungal suspension, and drench the roots with 200-300 ml per plant.

5. The planting method according to claim 1, characterized in that, The understory rotation planting includes: planting woody plants at a density of 3 meters between plants and 4 meters between rows; creating ridges between the rows of woody plants and planting 1-2 rows of Stephania tetrandra, wherein the distance between the Stephania tetrandra row and the woody plant row is not less than 1 meter, the row spacing of Stephania tetrandra is 60-80 cm, and the plant spacing is 30-40 cm.

6. The planting method according to claim 5, characterized in that, The woody plants selected are at least one of Camellia oleifera, walnut, Eucommia ulmoides, Magnolia officinalis, or Cunninghamia lanceolata, and are used for crop rotation.

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