Intercropping planting method for improving yield and quality of rhizoma atractylodis

Abstract

The invention relates to the technical field of agricultural product planting, in particular to an intercropping planting method for improving the yield and quality of rhizoma atractylodis, crops and rhizoma atractylodis are intercropped, the crops are used for regulating and controlling the abundance and diversity of rhizosphere microorganisms of rhizoma atractylodis, and the relative abundance and diversity of pathogenic bacteria are reduced; according to intercropping, crops and rhizoma atractylodis are intercropped according to the row ratio of 1: 2, the row spacing between the rhizoma atractylodis ranges from 25 cm to 35 cm, and the plant spacing ranges from 10 cm to 20 cm; the line spacing between the rhizoma atractylodis and the crops is 40 cm to 50 cm; the plant spacing between the crops is 30 cm to 40 cm; the crop is at least one of corn, wheat and millet. By setting the specific row spacing and plant spacing, the abundance and diversity of rhizoma atractylodis rhizosphere microorganisms are regulated and controlled, the relative abundance of pathogenic bacteria is reduced, continuous cropping obstacles are relieved, the rhizoma atractylodis yield is increased, the volatile oil content is increased, the medicinal material quality is effectively improved, ecological protection and economic benefits are considered, and the method is suitable for large-scale ecological rhizoma atractylodis planting.

Description

Technical Field

[0001] This invention relates to the field of agricultural product planting technology, specifically to an intercropping method for improving the yield and quality of Atractylodes lancea. Background Technology

[0002] Northern Atractylodes ( Atractylodes chinensis (DC.) Koidz. Atractylodes lancea (Cangzhu) is a perennial herbaceous plant belonging to the Asteraceae family. Its dried rhizome is used medicinally and is called Cangzhu. It is warm in nature and fragrant, with a slightly sweet, pungent, and bitter taste. It has the effects of drying dampness and strengthening the spleen, dispelling wind and turbidity, dispersing cold and improving eyesight, and is one of the commonly used Chinese medicinal materials. In recent years, the market size of Chinese medicinal materials has continued to expand, and Atractylodes lancea has shown strong development potential in the industry.

[0003] However, *Atractylodes lancea* grows slowly, requiring 5-6 years from sowing to harvest. With increasing cultivation years, its disease incidence and mortality rates rise significantly, especially with frequent outbreaks of white mold and root rot, leading to significant continuous cropping obstacles. Simultaneously, the problem of mixed germplasm is prominent, with multiple variants and even *Atractylodes lancea* often present in the same plot, severely impacting the quality of the medicinal material and planting efficiency. Wild resources have been severely depleted due to long-term over-harvesting and insufficient regeneration capacity, making artificial cultivation the main way to alleviate the supply-demand imbalance. However, monoculture often leads to poor growth, reduced yield, and lower quality of *Atractylodes lancea*. Using chemical fertilizers and pesticides to improve the yield and quality of *Atractylodes lancea* brings serious environmental problems. Long-term, excessive use of chemical fertilizers leads to soil acidification and compaction, damaging soil structure and reducing permeability and fertilizer retention capacity; pesticide residues pollute the soil and water systems, harm biodiversity, and threaten human health through the food chain.

[0004] Therefore, there is an urgent need to develop a cultivation method that can improve the yield and quality of Atractylodes lancea. Summary of the Invention

[0005] To address the above problems, this invention provides an intercropping method to improve the yield and quality of Atractylodes lancea.

[0006] This invention is achieved through the following technical solution: A method for intercropping Atractylodes lancea to improve yield and quality by regulating rhizosphere microorganisms is characterized by intercropping crops with Atractylodes lancea, using crops to regulate the abundance and diversity of rhizosphere microorganisms in Atractylodes lancea, thereby reducing the relative abundance and diversity of pathogenic bacteria.

[0007] Intercropping involves intercropping crops with Atractylodes lancea in a 1:2 row ratio. The row spacing between Atractylodes lancea plants is 25cm-35cm, and the plant spacing is 10cm-20cm. The row spacing between Atractylodes lancea plants and crops is 40cm-50cm. The plant spacing between crops is 30cm-40cm.

[0008] The crop is at least one of corn, wheat and millet.

[0009] Preferably, the Atractylodes is Northern Atractylodes (… Atractylodes chinensis (DC.) Koidz. ).

[0010] Preferably, the regulated microorganisms include at least one of the following categories: (1) Bacteria: Proteobacteria, Chloroflexi, Acidobacteriota, Actinobacteriota, WPS-2, Firmicutes, Gemmatimonadota.

[0011] (2) Fungi: Ascomycota, Mortierellomycota, Basidiomycota.

[0012] Preferably, the intercropping method can reduce the relative abundance of Ascomycota in the rhizosphere soil of Atractylodes lancea.

[0013] Preferably, improving the quality of Atractylodes lancea refers to: increasing the total concentration of volatile oil, increasing the content of atractylodes alcohol and / or increasing the content of β-cineole.

[0014] Preferably, the soil depth for planting is 0cm to 40cm.

[0015] Preferably, the soil pH value is 5.0~6.5.

[0016] Preferably, the growing season for the planted plants is from April to October each year.

[0017] Preferably, the fertilization management adopts a combination of organic and inorganic fertilizers, with a total nitrogen application rate of 180 kg / ha, a total phosphate fertilizer application rate of 225 kg / ha, and a total potassium fertilizer application rate of 105 kg / ha.

[0018] Compared with the prior art, the present invention has the following beneficial effects: An intercropping method for improving the yield and quality of Atractylodes lancea by regulating rhizosphere microorganisms involves intercropping Atractylodes lancea with other crops. The intercropping utilizes the crop to regulate the abundance and diversity of rhizosphere microorganisms in Atractylodes lancea, reducing the relative abundance and diversity of pathogenic bacteria. Specifically, the intercropping is performed with the other crops and Atractylodes lancea at a 1:2 row ratio. The row spacing between Atractylodes lancea plants is 25-35 cm, and the plant spacing is 10-20 cm. The row spacing between Atractylodes lancea and other crops is 40-50 cm. The plant spacing between other crops is 30-40 cm. The crops are at least one of corn, wheat, and millet. This invention achieves a systematic improvement from microbial regulation to growth environment optimization by employing an innovative 1:2 intercropping method with other crops and precise configuration of row and plant spacing. Its innovation lies in the first-time active regulation of the rhizosphere microbial community of Atractylodes lancea by co-planting crops such as corn, wheat, or millet, significantly reducing the relative abundance of pathogenic bacteria such as Ascomycota, thereby effectively alleviating continuous cropping obstacles. This method effectively increases the yield and biomass of Atractylodes lancea. When intercropped with corn, wheat, or millet at a 1:2 row ratio, the yield can be increased by 8%–12%, and the aboveground biomass also increases significantly, which is beneficial to the overall growth of the plant. Simultaneously, intercropping significantly improves the quality of Atractylodes lancea, especially increasing its volatile oil content. In wheat / Atractylodes lancea and millet / Atractylodes lancea treatments, the total volatile oil concentration increased by 45%–55%, with a significant increase in the content of key active ingredients such as atractylodes alcohol and β-cineole, meeting pharmacopoeia standards and ensuring the medicinal value of the herb. Furthermore, this method optimizes the growth traits of Atractylodes lancea, such as plant height and leaf area, enhancing photosynthetic capacity and plant vigor. In terms of soil ecology, intercropping regulates the rhizosphere microbial community structure, reduces the relative abundance of potential pathogens, and helps alleviate continuous cropping obstacles and maintain soil health. This method can also improve the utilization efficiency of resources such as light, water, and fertilizer, reduce environmental pressure, lower management costs, and achieve a balance between ecological and economic benefits, providing a feasible way for the ecological and standardized cultivation of Atractylodes lancea. Attached Figure Description

[0019] To more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the drawings used in the description of the embodiments or the prior art will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For those skilled in the art, other drawings can be obtained based on these drawings without creative effort.

[0020] Figure 1 This is a design drawing of the community according to the present invention.

[0021] Figure 2 This invention illustrates the effects of different intercropping methods on the yield and aboveground biomass of Atractylodes lancea at harvest time. Figure 2In the figure, (a) represents yield; (b) represents aboveground biomass; (c) represents leaf dry weight; and (d) represents stem dry weight. Note: Different letters in the figure represent different significant differences.

[0022] Figure 3 This invention illustrates the differences in functional traits of Atractylodes lancea at harvest time under different intercropping methods. Figure 3 In the figure, (a) represents plant height; (b) represents survival rate; (c) represents basal diameter; and (d) represents the number of buds. Note: Different letters in the figure represent different significant differences.

[0023] Figure 4 This invention relates to the effects of different intercropping methods on the volatile oil content of Atractylodes lancea at harvest time; Figure 4 In the figure, (a) represents the total harvest of the four volatile oils; (b) represents the total concentration of the four volatile oils; (c) represents the concentration of atractylone; (d) represents the concentration of atractylodes lancea; and (e) represents the concentrations of atractylodes lancea and β-eucalyptol in Atractylodes lancea. Note: Different letters in the figure represent different significant differences.

[0024] Figure 5 Non-metric multidimensional scale (NMDS) analysis of rhizosphere soil bacteria and fungi of Atractylodes lancea under different planting methods in this invention; Figure 5 In the table, (a) shows the non-metric multidimensional scale (NMDS) analysis of bacteria in the rhizosphere soil of Atractylodes lancea; and (b) shows the non-metric multidimensional scale (NMDS) analysis of fungi in the rhizosphere soil of Atractylodes lancea.

[0025] Figure 6 This invention provides a comparative analysis of the relative abundance of bacteria and fungi in the rhizosphere soil of Atractylodes lancea under different intercropping methods. Figure 6 In the table, (a) represents the relative abundance of bacteria; (b) represents the relative abundance of fungi.

[0026] Figure 7 This is a schematic diagram illustrating the effects of different intercropping methods of the present invention on the yield and quality of Atractylodes lancea. Detailed Implementation

[0027] To facilitate understanding of the present invention, a more comprehensive description is provided below, along with preferred embodiments. However, the present invention can be implemented in many different forms and is not limited to the embodiments described herein. Rather, these embodiments are provided to provide a thorough and complete understanding of the disclosure of the present invention.

[0028] Unless otherwise defined, all technical and scientific terms used in this invention have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains. The terminology used in this invention and in its specification is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention.

[0029] The beneficial effects of the present invention will be illustrated below through specific embodiments.

[0030] Example 1: An intercropping method to improve the yield and quality of Atractylodes lancea. Corn and Atractylodes lancea were intercropped at a row ratio of 1:2. The row spacing between Atractylodes lancea was 30cm and the plant spacing was 15cm. The row spacing between Atractylodes lancea and corn was 45cm and the plant spacing between corn was 34cm.

[0031] Example 2: An intercropping method to improve the yield and quality of Atractylodes lancea. Wheat and Atractylodes lancea were intercropped at a row ratio of 1:2. The row spacing between Atractylodes lancea was 30cm and the plant spacing was 15cm. The row spacing between Atractylodes lancea and wheat was 45cm and the plant spacing between wheat was 34cm.

[0032] Example 3: An intercropping method to improve the yield and quality of Atractylodes lancea. Millet and Atractylodes lancea were intercropped at a row ratio of 1:2. The row spacing between Atractylodes lancea plants was 30cm and the plant spacing was 15cm. The row spacing between Atractylodes lancea and millet was 45cm and the plant spacing between millet plants was 34cm.

[0033] Comparative Example 1: An intercropping method to improve the yield and quality of Atractylodes lancea. Corn and Atractylodes lancea were intercropped in a 2:2 row ratio. The row spacing between Atractylodes lancea was 30cm and the plant spacing was 15cm. The row spacing between Atractylodes lancea and corn was 45cm and the plant spacing between corn was 34cm.

[0034] Comparative Example 2: An intercropping method to improve the yield and quality of Atractylodes lancea. Corn and Atractylodes lancea were intercropped in a row ratio of 2:4. The row spacing between Atractylodes lancea was 30cm and the plant spacing was 15cm. The row spacing between Atractylodes lancea and corn was 45cm and the plant spacing between corn was 34cm.

[0035] Comparative Example 3: An intercropping method to improve the yield and quality of Atractylodes lancea. Soybeans and Atractylodes lancea were intercropped at a row ratio of 1:2. The row spacing between Atractylodes lancea was 30cm and the plant spacing was 15cm. The row spacing between Atractylodes lancea and soybeans was 45cm and the plant spacing between soybeans was 8.5cm.

[0036] Comparative Example 4: An intercropping method to improve the yield and quality of Atractylodes lancea. Marigolds and Atractylodes lancea were intercropped at a row ratio of 1:2. The row spacing between Atractylodes lancea was 30cm and the plant spacing was 15cm. The row spacing between Atractylodes lancea and marigolds was 45cm and the plant spacing between marigolds was 17cm.

[0037] Comparative Example 5: An intercropping method to improve the yield and quality of Atractylodes lancea. Peanuts and Atractylodes lancea were intercropped at a row ratio of 1:2. The row spacing between Atractylodes lancea was 30cm and the plant spacing was 15cm. The row spacing between Atractylodes lancea and peanuts was 45cm and the plant spacing between peanuts was 8.5cm.

[0038] Comparative Example 6: An intercropping method to improve the yield and quality of Atractylodes lancea. Intercropping of Coix lacryma-jobi and Atractylodes lancea was carried out at a row ratio of 1:2. The row spacing between Atractylodes lancea was 30cm and the plant spacing was 15cm. The row spacing between Atractylodes lancea and Coix lacryma-jobi was 45cm and the plant spacing between Coix lacryma-jobi was 17cm.

[0039] Comparative Example 7: An intercropping method to improve the yield and quality of Atractylodes lancea. Perilla and Atractylodes were intercropped in a row ratio of 1:2. The row spacing between Atractylodes was 30cm and the plant spacing was 15cm. The row spacing between Atractylodes and Perilla was 45cm and the plant spacing between Perilla was 17cm.

[0040] Comparative Example 8: An intercropping method to improve the yield and quality of Atractylodes lancea. Atractylodes lancea should be planted alone, with a row spacing of 30cm and a plant spacing of 15cm.

[0041] Comparative Example 9: An intercropping method to improve the yield and quality of Atractylodes lancea. When corn is planted as a monoculture, the row spacing between corn plants is 60cm and the plant spacing is 34cm.

[0042] It should be noted that in intercropping methods to improve the yield and quality of Atractylodes lancea, the row spacing between Atractylodes lancea should be 25cm-35cm, and the plant spacing should be 10cm-20cm; the row spacing between Atractylodes lancea and corn should be 40cm-50cm, the row spacing between Atractylodes lancea and wheat should be 40cm-50cm, and the row spacing between Atractylodes lancea and millet should be 40cm-50cm; the plant spacing between corn should be 30cm-40cm; the plant spacing between wheat should be 30cm-40cm or dense planting varieties should be used; and the plant spacing between millet should be 30cm-40cm or dense planting varieties should be used. All of these methods can improve the yield and quality of Atractylodes lancea.

[0043] Test case The following experiments were conducted on the planting methods in Examples 1-3 and Comparative Examples 1-9.

[0044] I. Complete Planting Steps: (1) Overview of the test site This invention was conducted from April to November 2024 at the Jilin Lishu Experimental Station of China Agricultural University (43.3°N, 124.4°E), located in Siping City, Jilin Province. This field experiment was established in 2023. The average temperature during the growing season was 17.7℃, the frost-free period ranged from 115 to 188 days, and the annual accumulated temperature (≥10℃) ranged from 2900℃ to 3100℃. This region is situated in the main maize-producing area of ​​Northeast China, with a temperate semi-arid climate. Annual precipitation ranges from 400 mm to 600 mm, and the precipitation distribution is uneven, exhibiting distinct monsoon characteristics. Rainfall from June to August accounts for 65% of the total annual precipitation, indicating a concentrated rainy season. The average annual evaporation reaches 808 mm, and the aridity index is approximately 1.35, indicating a water deficit in this region. In addition, the area has abundant solar energy resources, with an annual sunshine duration of 2644.2 hours, an average of about 8 hours of sunshine per day, and an annual total solar radiation of 191.28 kcal / cm², of which photosynthetically active radiation (PAR) is as high as 247.43 kcal / cm², providing ample energy for crop photosynthesis. The basic physicochemical properties of the soil in the Atractylodes lancea diversified planting experimental plots are shown in Table 1.

[0045] Table 1. Basic physical and chemical properties of soil in the experimental plots for diversified cultivation of Atractylodes lancea (2) Screening Experiment Design A single-factor experimental design was used, setting up 12 planting methods: maize / Atractylodes lancea intercropping (row ratio 1:2), maize / Atractylodes lancea intercropping (row ratio 2:2), maize / Atractylodes lancea intercropping (row ratio 2:4), millet / Atractylodes lancea intercropping (row ratio 1:2), soybean / Atractylodes lancea intercropping (row ratio 1:2), marigold / Atractylodes lancea intercropping (row ratio 1:2), wheat / Atractylodes lancea intercropping (row ratio 1:2), peanut / Atractylodes lancea intercropping (row ratio 1:2), Job's tears / Atractylodes lancea intercropping (row ratio 1:2), perilla / Atractylodes lancea intercropping (row ratio 1:2), Atractylodes lancea monoculture, and maize monoculture. There were 12 treatments, with three replicates, totaling 36 plots. Figure 1 As shown.

[0046] All residential areas are arranged in an east-west direction. Each area is 6m long and 7.5m wide, with a total area of ​​6 × 7.5m = 45m². 2 Within the planting area, plants are oriented east-west. The row spacing and plant spacing between Atractylodes lancea plants are 30cm and 15cm respectively, and the row spacing between Atractylodes lancea plants and adjacent plants is 45cm. Wheat and millet are planted densely, corn plants are spaced 34cm apart, Job's tears, marigolds, and perilla plants are spaced 17cm apart, and soybeans and peanuts are spaced 8.5cm apart. The east-west spacing within each planting area is 0.5m, the north-south spacing is 0.6m, and each repeating area has a 0.5m interval.

[0047] Intercropping maize / Atractylodes lancea in a 1:2 row ratio, as well as intercropping other plants besides maize, all use a 1:2 row ratio, with a total of 3 strips. Each strip includes four rows of Atractylodes lancea and two rows of other plants. Intercropping maize / Atractylodes lancea in a 2:2 row ratio uses a total of 4 strips, with each strip including two rows of Atractylodes lancea and one row of maize.

[0048] (3) Fertilizer management: adopt the method of applying organic and inorganic fertilizers together.

[0049] For monoculture of Atractylodes lancea, monoculture of maize, and intercropping of Atractylodes lancea with non-leguminous crops: the total nitrogen application rate is 180 kg / ha (75 kg / ha of organic fertilizer and 105 kg / ha of inorganic fertilizer). Base fertilizer application is 75 kg / ha of organic fertilizer and 80 kg / ha of inorganic fertilizer. Topdressing with inorganic fertilizer at the maize jointing stage is 25 kg / ha.

[0050] Intercropping Atractylodes lancea / Leguminates: Nitrogen application rate is 167.5 kg / ha (75 kg / ha of organic fertilizer and 92.5 kg / ha of inorganic fertilizer). Apply 75 kg / ha of organic fertilizer and 80 kg / ha of inorganic fertilizer as base fertilizer, and apply 12.5 kg / ha of inorganic fertilizer as top dressing during the corn jointing stage.

[0051] Phosphate fertilizer: The total application rate is 225 kg / ha (97 kg / ha of organic fertilizer and 128 kg / ha of inorganic fertilizer), all of which are applied as base fertilizer.

[0052] Potassium fertilizer: The total application rate is 105 kg / ha (all organic fertilizer), and it is applied as base fertilizer.

[0053] II. Samples and Collection Methods Atractylodes samples: The seedlings were transplanted from two-year-old Atractylodes lancea plants, all from Jianyou Atractylodes lancea Planting Professional Cooperative in Qinglong Manchu Autonomous County, Qinhuangdao City, Hebei Province. They were identified as Atractylodes lancea by Associate Professor Qiao Xu of the Institute of Medicinal Plants. A. chinensis Authentic product.

[0054] The remaining paired crops are all common varieties native to Jilin.

[0055] The period from late April to early October is the plant growth period (April to August for wheat). During this period, samples were taken from the paired plants and Atractylodes lancea in each plot once during the vigorous growth period (August) and once during the harvest period (October), for a total of 2 samplings (wheat was sampled only once in August).

[0056] Sampling of Atractylodes lancea: During the harvest season, ten plants of Atractylodes lancea from each plot were sampled to measure functional traits (plant height, basal diameter, number of buds, etc.); during the yield measurement, all plants in the sampling zone were sampled, avoiding the plants being tested, and the stems, leaves and rhizomes of all plants within the last 4 meters were sampled to measure biomass and productivity.

[0057] Rhizosphere soil was collected during the vigorous growth period. In the sampling areas of the experimental plots of Atractylodes lancea monoculture, millet and Atractylodes lancea intercropping (1:2), wheat and Atractylodes lancea intercropping (1:2), and maize and Atractylodes lancea intercropping (1:2), the "root shaking method" was used to select 5 Atractylodes lancea plants for sampling. The entire root of Atractylodes lancea was carefully dug out, and large clumps of soil attached to the root were shaken off. The soil attached to the root was quickly brought back to the laboratory and collected. Root stubble, stones and other impurities were removed. The soil from the rhizosphere of the 5 Atractylodes lancea plants was mixed to form 12 soil samples, which were stored in a -20℃ refrigerator for testing.

[0058] III. Sample Preparation and Measurement (1) Biomass determination (fresh and dry weight of aboveground and underground parts): After cleaning the stems, leaves and roots of Atractylodes lancea, wipe off the excess water on the surface, and then weigh the fresh weight of the aboveground and underground parts of Atractylodes lancea using a balance of 0.01%. After weighing, place the stems, leaves and roots of Atractylodes lancea on kraft paper and put them in a drying room to air dry naturally. After about two weeks, weigh the dry weight of the samples for 3 consecutive days. If the measured value remains unchanged, record it as the dry weight of the roots, stems and leaves of Atractylodes lancea. After weighing, put them into an envelope.

[0059] (2) Determination of nutrient content in plants: Naturally air-dried Atractylodes lancea samples were taken, and the roots, stems, and leaves were ground into powder using a pulverizer and passed through a 60-mesh sieve. 0.3500g~0.4000g of plant samples were accurately weighed using a 0.01% balance, transferred to test tubes, and 5mL of concentrated sulfuric acid was added. The mixture was left to stand overnight. The next day, digestion was performed. The digestion furnace was heated to 270℃ for 30min; then the temperature was increased to 360℃ for 2h~3h. During digestion, 2mL of hydrogen peroxide solution was added to the test tubes three times. The digestion was complete when the liquid in the test tube became clear and transparent. After the digestion solution was cooled to room temperature, the volume was adjusted to 75mL. The liquid was then used for the determination of nitrogen, phosphorus, and potassium in the plants. The Kjeldahl method was used to determine the nitrogen content, the vanadium-molybdenum yellow colorimetric method was used to determine the phosphorus content, and the flame photometry method was used to determine the potassium content.

[0060] (3) Determination of volatile oil content: Weigh 0.8 g of sample, add 20 mL of methanol, vortex, and extract by ultrasonication at 40℃ for 60 min. Centrifuge at 5000 r / min for 10 min, take the supernatant, dilute it 10 times with acetonitrile-water (1:1), filter it through a 0.22 μm filter membrane, and then load it into the chromatographic instrument. Chromatographic conditions: chromatographic column: Symmetry C18 5 μm 4.6 mm × 150 mm, mobile phase A: acetonitrile, mobile phase B: 0.1% phosphoric acid water (w / v), flow rate: 1 mL / min, column temperature: 30℃, detection wavelength: 203 nm, injection volume: 20 μL.

[0061] (4) Soil microbial abundance and diversity: Soil samples were flash-frozen in liquid nitrogen and stored at -80℃. Total microbial DNA was extracted using the MoBio DNA extraction kit and MP homogenizer. After the extracted nucleic acids passed quality control by an ultra-micro spectrophotometer and a bioanalyzer, PCR amplification of the 16S rRNA gene V3-V4 hypervariable region (primer 338F / 806R) or fungal ITS sequence was performed using a 96-well gradient PCR instrument for 30-35 cycles. The amplified products were sequenced at 2×250bp paired ends using a sequencing platform. The raw data were imported into QIIME2 software for quality control, noise reduction, and ASV clustering. Species annotation was performed using the Silva database, and α diversity index and β diversity based on Bray-Curtis distance were calculated using R language. The absolute abundance of microorganisms was accurately quantified by 16S rRNA gene copy number using an ABI 7500 real-time quantitative PCR system.

[0062] IV. Results and Analysis Indicator 1: Biomass.

[0063] Different intercropping methods had no significant impact on the yield of Atractylodes lancea at harvest time (October). Intercropping Atractylodes lancea with millet, wheat, and peanuts showed an increasing trend, increasing yield by 8%–12%. Intercropping Atractylodes lancea with perilla and corn at row ratios of 1:2 and 2:4 resulted in yields that were basically the same as monocultures. Intercropping Atractylodes lancea with soybeans, marigolds, coix seeds, and corn at a 2:2 ratio showed a decreasing trend in yield. Specifically, intercropping Atractylodes lancea with marigolds reduced yield by 49%. Figure 2 As shown.

[0064] Different intercropping methods affect the aboveground biomass of Atractylodes lancea at harvest. Intercropping with millet, wheat, and maize (1:2) tends to increase the aboveground biomass of Atractylodes lancea at harvest. However, intercropping with marigolds significantly reduces the aboveground biomass of Atractylodes lancea. The aboveground biomass of other intercropping methods is not significantly different from that of monoculture. Different intercropping methods have a significant impact on the leaf dry weight and stem dry weight of Atractylodes lancea at harvest. P <0.05). Among them, the dry weight of leaves increased by 33.6% and the dry weight of stems increased by 72.0% when millet was intercropped with Atractylodes lancea; when intercropped with marigolds, the dry weight of leaves of Atractylodes lancea decreased by 80.9% and the dry weight of stems decreased by 67.8%. Figure 2 As shown.

[0065] Indicator 2: Functional traits.

[0066] During the harvest period, intercropping with peanuts, millet, wheat, coix seed, and corn at a 1:2 ratio all tended to increase the plant height of Atractylodes lancea. Compared with monoculture, intercropping with marigolds, except for marigolds, did not decrease the plant height. Buds refer to the hairy buds attached to the round stem base of Atractylodes lancea roots. Atractylodes lancea stems are thin, fragile, and easily broken. Buds can break through the soil layer and grow into stems after stem breakage, allowing Atractylodes lancea to continue growing. A higher number of buds indicates a higher survival rate in the later stages. Results showed that intercropping with marigolds, coix seed, and corn significantly reduced the number of buds in Atractylodes lancea. However, different intercropping methods had no significant effect on the survival rate and basal diameter of Atractylodes lancea. Figure 3 As shown.

[0067] Index 3: Volatile oil content.

[0068] Analysis of the volatile oil concentration of Atractylodes lancea at harvest revealed that different intercropping methods had no significant impact on the total volatile oil yield of the four types of Atractylodes lancea. However, intercropping with millet, wheat, peanut, marigold, perilla, corn (1:2), and corn (2:4) showed an increasing trend in the total volatile oil yield of the four types of Atractylodes lancea. Figure 4 As shown in (a) above. All intercropping methods increased the total concentration of the four volatile oils in Atractylodes lancea, with the ratios being wheat / Atractylodes lancea > millet / Atractylodes lancea > marigold / Atractylodes lancea > perilla / Atractylodes lancea > peanut / Atractylodes lancea > soybean / Atractylodes lancea > corn / Atractylodes lancea (1:2) > corn / Atractylodes lancea (2:2) > coix seed / Atractylodes lancea > corn / Atractylodes lancea (2:2). Among these, intercropping with perilla, marigold, millet, and wheat significantly increased the total volatile oil concentration of Atractylodes lancea by 45%–55%. P <0.05), such as Figure 4 As shown in (b) of the diagram. Compared with monoculture of Atractylodes lancea, intercropping with soybeans increased the concentration of atractylone in Atractylodes lancea, while wheat decreased the concentration of atractylone. Other intercropping methods had no significant effect on the concentration of atractylone in Atractylodes lancea, such as... Figure 4 As shown in (c), different intercropping methods significantly affected the concentrations of atractylodes lancea and β-cineole in Atractylodes lancea. Intercropping with wheat and millet significantly increased the concentrations of atractylodes lancea and β-cineole. Intercropping with peanuts, marigolds, perilla leaves, coix seeds, and corn at 1:2 and 2:2 ratios also increased the concentrations of these two substances in Atractylodes lancea. Figure 4 As shown in (e) in the figure. Experiments revealed that different intercropping methods did not significantly affect the concentration of atractylodes in Atractylodes lancea at harvest time. However, except for intercropping with soybeans and coix seeds, the concentration of atractylodes in Atractylodes lancea under other intercropping methods met the pharmacopoeia requirements, such as... Figure 4 As shown in (d) in the figure.

[0069] Indicator 4: Soil microbial abundance and diversity.

[0070] Intercropping altered the relative abundance and community structure of fungi in the rhizosphere soil of Atractylodes lancea. Intercropping with wheat and Atractylodes lancea, as well as with maize and Atractylodes lancea (1:2), significantly reduced the relative abundance of Ascomycota in the rhizosphere soil of Atractylodes lancea, while intercropping with millet and wheat significantly altered the fungal community structure in the rhizosphere soil of Atractylodes lancea.

[0071] The differences in rhizosphere soil bacterial communities of Atractylodes lancea under different intercropping methods were not significant, such as Figure 5 As shown, there is significant overlap between the groups in the NMDS plot, indicating that the microbial community composition of these samples is very similar and difficult to distinguish. Figure 5 It can be seen that the fungal community structure differed significantly between monoculture of Atractylodes lancea and the rice / Atractylodes lancea and corn / Atractylodes lancea (1:2) ratios. Intercropping altered the rhizosphere soil microbial community of Atractylodes lancea, with significant differences in fungal community structure between the rice / Atractylodes lancea and corn / Atractylodes lancea (1:2) ratios.

[0072] OTUs were aggregated at the phylum level, and the ratio of phylum-related microbial abundance to total OTU abundance was calculated. Based on species annotation results, it was found that the bacterial phyla with the highest relative abundance were the same across four cultivation methods: Atractylodes lancea monoculture, millet / Atractylodes lancea, wheat / Atractylodes lancea, and maize / Atractylodes lancea. Figure 6 As shown. Proteobacteria ( ) in the rhizosphere soil of Atractylodes lancea under different planting methods. Proteobacteria ), Green Curvature ( Chloroflexi ), Acidobacteria ( Acidobacteriota ), Actinobacteria ( Actinobacteriota The phylum with relatively high abundance is . The phylum with relatively low abundance is Pseudobacteria ( WPS -2), Firmicutes ( Firmicutes )Bacillus phylum ( Gemmatimonadota At the fungal level, the relative abundance of rhizosphere fungi in Atractylodes lancea varied significantly under different planting methods. The relative abundance of Ascomycota in the rhizosphere soil of monoculture Atractylodes lancea was significantly higher than that of intercropping with millet / Atractylodes lancea and wheat / Atractylodes lancea. P <0.05), other dominant phyla such as Zygomycetes ( Mortierellomycota Basidiomycota ( Basidiomycota There was no significant difference in relative abundance. P >0.05).

[0073] V. Summary This invention designed a randomized block experiment, selecting twelve planting methods (Atractylodes lancea monoculture, maize monoculture, maize / Atractylodes lancea (1:2, 2:2, 2:4), millet / Atractylodes lancea, wheat / Atractylodes lancea, marigold / Atractylodes lancea, soybean / Atractylodes lancea, peanut / Atractylodes lancea, coix seed / Atractylodes lancea, and perilla / Atractylodes lancea) to explore the effects of different intercropping systems on the yield and quality of Atractylodes lancea. The main conclusions are as follows:

[0074] (1) Intercropping can increase the yield and biomass of Atractylodes lancea. Among them, the yield of Atractylodes lancea is higher when intercropped with millet, wheat, peanut, and corn (1:2). The aboveground biomass accumulation of Atractylodes lancea is greater when intercropped with millet / Atractylodes lancea, wheat / Atractylodes lancea, and corn / Atractylodes lancea (1:2). The yield and biomass of the Atractylodes lancea / corn intercropping system are greater than those of monoculture, and the yield LER and biomass LER are both greater than 1, which proves that intercropping has a yield advantage. In general, corn / Atractylodes lancea (1:2) intercropping is the most suitable grain-medicinal herb intercropping method for widespread promotion.

[0075] (2) Intercropping improved the functional traits of Atractylodes lancea. After intercropping at harvest, the plant height of Atractylodes lancea increased, and the leaf area of ​​some intercropped plants (corn / Atractylodes lancea (1:2), marigold / Atractylodes lancea, coix seed / Atractylodes lancea, wheat / Atractylodes lancea) increased, which promoted light interception of Atractylodes lancea.

[0076] (3) Intercropping methods promoted the increase of Atractylodes lancea volatile oil concentration. Overall, wheat / Atractylodes lancea, millet / Atractylodes lancea, marigold / Atractylodes lancea, and perilla / Atractylodes lancea intercropping significantly increased the total volatile oil concentration of Atractylodes lancea. Compared with other methods, the concentration of atractylodes lancea was higher under corn / Atractylodes lancea (1:2) intercropping. The concentrations of atractylol and β-eucalyptol in Atractylodes lancea were significantly increased under wheat / Atractylodes lancea and millet / Atractylodes lancea intercropping.

[0077] (4) Intercropping affects the relative abundance of bacterial and fungal species and community structure in the rhizosphere soil of Atractylodes lancea. Different intercropping methods have no significant effect on the bacterial species diversity and community structure in the rhizosphere soil of Atractylodes lancea, but have a significant effect on the level of fungal species diversity and community structure. Among them, wheat / Atractylodes lancea and maize / Atractylodes lancea (1:2) intercropping methods have a greater advantage in improving the soil microenvironment and can be further optimized and promoted as ecological planting methods for Atractylodes lancea.

[0078] In conclusion, diversified planting can promote the improvement of Atractylodes lancea quality. Wheat / Atractylodes lancea, millet / Atractylodes lancea, and corn / Atractylodes lancea (1:2) can increase the volatile oil concentration of Atractylodes lancea without reducing yield, making them suitable intercropping methods for demonstration and promotion. Figure 7 As shown.

[0079] The technical features of the above embodiments can be combined in any way. For the sake of brevity, not all possible combinations of the technical features in the above embodiments are described. However, as long as there is no contradiction in the combination of these technical features, they should be considered to be within the scope of this specification.

[0080] The embodiments described above are merely illustrative of several implementations of the present invention, and while the descriptions are specific and detailed, they should not be construed as limiting the scope of the invention. Those skilled in the art can make various modifications and improvements without departing from the concept of the present invention, and these all fall within the scope of protection of the present invention. Therefore, the scope of protection of this invention should be determined by the appended claims.

Claims

1. An intercropping method for improving the yield and quality of Atractylodes lancea by regulating rhizosphere microorganisms, characterized in that, Intercropping crops with Atractylodes lancea can help regulate the abundance and diversity of rhizosphere microorganisms in Atractylodes lancea and reduce the relative abundance and diversity of pathogens. Intercropping involves intercropping crops with Atractylodes lancea in a 1:2 row ratio. The row spacing between Atractylodes lancea plants is 25cm-35cm, and the plant spacing is 10cm-20cm. The row spacing between Atractylodes lancea plants and crops is 40cm-50cm. The plant spacing between crops is 30cm-40cm. The crop is at least one of corn, wheat and millet.

2. The intercropping method as described in claim 1, characterized in that, The Atractylodes lancea mentioned is Northern Atractylodes lancea ( Atractylodes chinensis (DC.) Koidz. ).

3. The intercropping method as described in claim 1, characterized in that, The regulated microorganisms include at least one of the following categories: (1) Bacteria: Proteobacteria, Chloroflexi, Acidobacteriota, Actinobacteriota, WPS-2, Firmicutes, Gemmatimonadota. (2) Fungi: Ascomycota, Mortierellomycota, Basidiomycota.

4. The intercropping method as described in claim 3, characterized in that, The intercropping method described above can reduce the relative abundance of Ascomycota in the rhizosphere soil of Atractylodes lancea.

5. The intercropping method as described in claim 1, characterized in that, The improvement of Atractylodes lancea quality refers to: increasing the total concentration of volatile oil, increasing the content of atractylodes alcohol and / or increasing the content of β-cineole.

6. The intercropping method as described in claim 1, characterized in that, The soil depth for planting is 0cm to 40cm.

7. The intercropping method as described in claim 1, characterized in that, The soil pH value is 5.0~6.

5.

8. The intercropping method as described in claim 1, characterized in that, The growing season for the plants is from April to October each year.

9. The intercropping method as described in claim 1, characterized in that, Fertilization management adopts a combination of organic and inorganic fertilizers, with a total nitrogen application rate of 180 kg / ha, a total phosphate fertilizer application rate of 225 kg / ha, and a total potassium fertilizer application rate of 105 kg / ha.

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