Method for old ginseng hole continuous cropping of ginseng

Abstract

The present application relates to the field of Gastrodia elata cultivation, and particularly relates to a method for continuously planting Gastrodia elata in old planting holes. The present application effectively improves the yield of Gastrodia elata, and significantly reduces the incidence of new Gastrodia elata and the decay rate of mother Gastrodia elata by scientifically planting after rhizoma polygonati. In the present application, the breeding hole is sexual reproduction, and it does not need to cultivate armillaria bed. The armillaria, fungus material and Gastrodia elata seeds can be placed at the same time. The planting hole is asexual reproduction. Since the Gastrodia elata seed needs a large amount of nutrient elements, the armillaria bed needs to be cultivated. This operation mode can form the difference of microbial community between the breeding hole and the planting hole, which is beneficial to ecological competition, and thus improves the soil recovery and improvement.

Description

Technical Field

[0001] This invention relates to the field of Gastrodia elata cultivation, specifically to a method for continuous planting of Gastrodia elata in existing planting holes. Background Technology

[0002] Gastrodia elata ( Gastrodia elata Blume Gastrodia elata is a perennial heterotrophic herb belonging to the genus Gastrodia elata in the Orchidaceae family. Its dried underground tubers are used as medicine and are a traditional and precious Chinese medicinal material with effects such as calming liver yang and dispelling wind and unblocking collaterals.

[0003] Currently, semi-wild cultivation under forest cover is one of the main production models for Gastrodia elata in our province. However, Gastrodia elata suffers from continuous cropping obstacles, meaning that after one crop of semi-wild Gastrodia elata is planted under forest cover, the land needs to be left idle for more than four years before it can be planted again. This not only leads to problems such as high planting costs, unstable yields, and land abandonment, but also, with the extension of the history of large-scale cultivation, the land resources available for Gastrodia elata cultivation are becoming increasingly scarce, which will seriously threaten the healthy and sustainable development of the Gastrodia elata industry. Therefore, developing a green, sustainable Gastrodia elata continuous cropping technology with industrial application prospects is of great practical significance for solving the problems of long soil resource utilization cycles and low sustainable production capacity of Gastrodia elata cultivation.

[0004] The essence of crop rotation obstacles in Gastrodia elata lies in the fact that the changes in the soil environment caused by the growth of the previous crop exceed its own adaptability. These changes are mainly manifested in soil nutrient imbalance, enrichment of soil pathogens, and accumulation of allelopathic substances. Among these, the secretions of Gastrodia elata, the metabolites of Armillaria mellea, and the decomposition products of the fungal material all have certain allelopathic effects and are important reasons for continuous cropping obstacles in Gastrodia elata. The imbalance of soil microecology, especially the increase of pathogenic fungi of the genus *Solanum lyratum*, is the fundamental reason for the high incidence of diseases in continuously cropped Gastrodia elata. Crop rotation or intercropping is a common method in this field to overcome continuous cropping obstacles. For example, CN116114555A discloses a method for semi-wild cultivation of Polygonatum in old ponds for Gastrodia elata, which clarifies that Polygonatum seedlings are harvested three years after transplantation, and the land after harvest can be directly used for Gastrodia elata cultivation. Another example is CN112005832A, which discloses a method for rotating and intercropping Gastrodia elata and Polygonatum sibiricum, which clarifies that Gastrodia elata can be cultivated after Polygonatum sibiricum cultivation. Yet another example is CN115039650A, which discloses a method for rotating Gastrodia elata and Polygonatum sibiricum, which clarifies that Gastrodia elata is planted after Polygonatum sibiricum is harvested. However, the technical solution of CN116114555A has a long harvest time for Polygonatum sibiricum and focuses on the quality of Polygonatum sibiricum; the technical solution of CN112005832A only focuses on the yield of Polygonatum sibiricum and Gastrodia elata, and cannot guarantee the quality; the technical solution of CN115039650A only focuses on the yield and quality of Polygonatum sibiricum, and cannot guarantee the quality of Gastrodia elata. In summary, existing technologies only demonstrate that crop rotation between Gastrodia elata and Polygonatum sibiricum can improve land utilization. However, it is unknown whether Polygonatum sibiricum can neutralize allelopathic substances produced by the previous crop of Gastrodia elata, restore soil physicochemical properties, nutrient and microbial community structure, and ensure the yield and quality of subsequent Gastrodia elata. Furthermore, since the soil in the planting holes left after Gastrodia elata harvest is often too moist, it is very easy for Polygonatum sibiricum root rot to occur. Whether it will have an adverse effect on the growth of Armillaria mellea, and whether it will cause cross-infection of diseases and pests in subsequent Gastrodia elata crops, are all unknown. Therefore, in order to ensure the quality and yield of continuous cropping of Gastrodia elata, the inventors, based on practical planting experience and planting theory, have provided a method for continuous cropping of Gastrodia elata in old planting holes. Summary of the Invention

[0005] To ensure the yield and quality of continuously cropped Gastrodia elata, this invention provides a method for continuously cropping Gastrodia elata in old planting holes by improving the soil microenvironment for Armillaria mellea growth.

[0006] The technical solution of the present invention: A method for treating Gastrodia elata in old acupoints repeatedly includes the following steps: Step 1: Propagation of Gastrodia elata: From March to July of the first year, select suitable wasteland for planting Gastrodia elata, dig breeding holes and loosen the soil. Then, lay a layer of fungal material with a length of 10-20cm and a spacing of ≥2cm. Fill the hole with clean, fine soil until 1 / 3 of the fungal material surface is exposed. Place Armillaria mellea and mixed with germinated Gastrodia elata seeds evenly at both ends and sides of the fungal material. Cover with clean, fine soil until the fungal material is no longer visible. Then lay another layer of fungal material, place Armillaria mellea and Gastrodia elata seeds on top, cover with soil, and then cover with fallen leaves. The thickness of the fallen leaves should be 8-12cm to help keep the planting hole warm and moist. Step 2: Cultivating the mycelium bed: After the breeding is completed, dig planting holes and loosen the soil at a distance from the breeding holes. Then, lay a layer of mycelium material with a length of 10-20cm and a spacing of ≥2cm. Fill the hole with clean, fine soil until 1 / 3 of the mycelium surface is exposed. Place the Armillaria mellea evenly at both ends and sides of the mycelium material and cover it with clean, fine soil until the mycelium material is no longer visible. Then, lay another layer of mycelium material, place the Armillaria mellea and Gastrodia elata seeds on top, cover with soil, and then cover with fallen leaves. The thickness of the fallen leaves covering should be 8-12cm. Step 3: Planting Polygonatum: From October to December of the second year, dig up the propagated Gastrodia elata seeds and select them to obtain Gastrodia elata seeds; then collect the fungal material in the breeding holes and screen out the incompletely decomposed parts, crush them to obtain 2-3cm fragments, and spread them evenly in the Gastrodia elata breeding holes. At the same time, add quicklime to the Gastrodia elata breeding holes, plow to 15-20cm, cover the surface with the excavated soil, then prepare the land and make ridges. Sow Polygonatum in March of the year following the harvest, using disease-free tubers as seeds. After sowing, cover with soil and cover with fallen leaves or straw from the forest. Step Four: Planting Gastrodia elata and Polygonatum sibiricum: Select the chosen Gastrodia elata seeds and inoculate them onto the mycelium bed. Harvest the Gastrodia elata in November-December of the third year. Then collect the mycelium material from the planting holes, sifting out any incompletely decomposed parts. Crush the incompletely decomposed material to obtain 2-3cm fragments, and evenly spread them into the Gastrodia elata planting holes. Simultaneously, add quicklime to the planting holes, till the soil to a depth of 15-20cm, and cover the surface with the excavated soil. Then, prepare the land by ridging. Sow Polygonatum sibiricum in March of the following year, using disease-free tubers as seeds. After sowing, cover with soil, then with fallen leaves or straw from the forest floor. Step 5: Cyclic planting: After planting Polygonatum for 2-5 years, harvest it. After harvesting, turn over the remaining stems, leaves and fine roots on the spot and return them to the field. Then, follow the methods of Step 1 to Step 4 to lay out new fungal material and plant Gastrodia elata seeds.

[0007] In step one, fresh twigs with no obvious fungal spots are placed between the Armillaria mellea and the germinated Gastrodia elata seeds to promote the branching growth of the Armillaria mellea mycelium.

[0008] In step four, the inoculation includes the following steps: 1) First, remove the covering material and soil from the surface of the fungal substrate, take out the fungal substrate placed on the surface, and then continue to dig down to the bottom layer of fungal substrate. Pull out 1 / 3 of the substrate surface, and evenly place 1 / 2 of the seed spawn at both ends and sides of the fungal substrate. Then cover it with clean fine soil until the seed spawn is no longer visible. 2) Arrange the removed fungal materials at intervals of ≥2cm, fill them with clean fine soil, leaving 1 / 3 of the material exposed, place the remaining seeds evenly at both ends and sides of the fungal material, cover them with fine soil until the seeds are no longer visible, cover with soil 1-3cm, and finally cover with fallen leaves.

[0009] The beneficial effects of this invention are: This invention effectively increases the yield of Gastrodia elata and significantly reduces the incidence of disease and seed rot rate of new Gastrodia elata by scientifically rotating it with Polygonatum sibiricum before replanting.

[0010] This invention utilizes a rotation system of *Polygonatum sibiricum* in both breeding and planting holes, achieving full utilization of land resources. It also effectively reduces soil pH, the content of available nitrogen and phosphorus, and significantly increases the activity of polyphenol oxidase and laccase in the soil, as well as the quantity and species diversity of soil microorganisms. This provides a favorable environment for subsequent sexual and asexual cultivation of *Gastrodia elata*. Furthermore, the rotation treatment improves soil fertility indicators and provides favorable conditions for the robust growth of *Armillaria mellea*. Crop rotation increases the number and diversity of soil microorganisms. The rotation model and hole-opening method of this invention also significantly reduces the abundance of common pathogenic fungi such as *Cladosporium* and *Armillaria* in the soil after rotating with *Polygonatum sibiricum*. This not only effectively inhibits the incidence of disease in *Gastrodia elata* but also provides a favorable environment for the growth of *Armillaria mellea*.

[0011] In this invention, the breeding pit is for sexual reproduction, which does not require the cultivation of Armillaria mellea mycelium bed. It can achieve the simultaneous placement of Armillaria mellea, mycelium material and Gastrodia elata seed. The planting pit is for asexual reproduction. Since Gastrodia elata seed requires a large amount of Armillaria mellea mycelium to provide nutrients, mycelium bed cultivation is necessary. This operation method can create a difference in microbial communities between the breeding pit and the planting pit, which is conducive to ecological competition, thereby restoring and improving the soil microenvironment and providing a good growth environment for improving the yield and quality of Gastrodia elata.

[0012] In this invention, only a 3-year rotation of Polygonatum sibiricum is required before Gastrodia elata can be planted again, which greatly shortens the continuous cropping time of Gastrodia elata. Attached Figure Description

[0013] Figure 1 Results of the influence of each treatment group on the soil enzyme activity in the Gastrodia elata pits in Experiment Example 1.

[0014] Figure 2 Schematic diagram of soil fungal OUTs composition in each treatment group in Experiment Example 1.

[0015] Figure 3The results of the relative richness distribution of soil fungal community composition at the phylum and genus levels in each treatment group in Experiment Example 1, where a represents the phylum level and b represents the genus level.

[0016] Figure 4 Results of Beta diversity analysis of soil fungal communities at the genus level in each treatment group in Experiment Example 1. Detailed Implementation

[0017] The specific embodiments of the present invention will be described in further detail below, but the present invention is not limited to these embodiments. Any improvements or substitutions based on the basic spirit of these embodiments shall still fall within the scope of protection claimed by the claims of the present invention.

[0018] Experimental Site: A semi-wild cultivation base for Gastrodia elata in Dafang County, Bijie City, Guizhou Province: Altitude approximately 1200-1900m, central coordinates (105°36'629″E, 27°20'816″N). It has a subtropical humid monsoon climate, characterized by mild temperatures and abundant rainfall. The average annual temperature is around 11.8℃, with the coldest month (January) averaging 1.6℃ and the hottest month (July) averaging 20.7℃. The average annual rainfall is 1150.4mm. The slope ranges from 20° to 37°. The dominant vegetation consists of broad-leaved mixed forests such as oak and birch, and the soil type is mountain sandy loam.

[0019] Testing instruments: High performance liquid chromatograph (Waters e2695), inductively coupled plasma atomic emission spectrometer (Optima 8100); Kjeldahl nitrogen analyzer (Kjeltec 8400), centrifuge (Thermo Scientific), polymerase chain reaction (PCR) system (9700), 1 / 1000 electronic balance (YP5001), 0.001% balance (JA2003N), digital display constant temperature water bath (HH-6), spectrophotometer (UV-2600), grinder (FW177), muffle furnace (SX-12-10), MISEQ sequencer (Illumina Miseq), shaking incubator (RAH-250Y), etc.

[0020] Hemp Seeds: Select high-quality white hemp seeds with no surface diseases or mechanical damage, and the seed weight should be more than 3g.

[0021] Fungal substrate: Guizhou oak, birch, etc.

[0022] Example 1: A method for treating Gastrodia elata in old acupoints repeatedly includes the following steps: Step 1: Propagation of Gastrodia elata: From March to July of the first year, select suitable wasteland for planting Gastrodia elata, dig breeding holes and loosen the soil. The specifications of the breeding holes are 80cm (length) * 60cm (width) * 15cm (depth). Then, lay a layer of 15cm long mycelium material at a spacing of about 2cm. Fill the hole with clean, fine soil until 1 / 3 of the mycelium material surface is exposed. Place Armillaria mellea and germinated Gastrodia elata seeds evenly at both ends and sides of the mycelium material. Place fresh branch nodes without obvious mycelial spots between the Armillaria mellea and germinated Gastrodia elata seeds to promote the branching growth of Armillaria mellea mycelium. Cover with clean, fine soil until the mycelium material is no longer visible. Then lay another layer of mycelium material, then another layer of mycelium material, and then place the Armillaria mellea and Gastrodia elata seeds. Cover with soil and then cover with about 10cm of fallen leaves to help retain heat and moisture.

[0023] Step 2: Cultivating the substrate: After the breeding is completed, dig planting holes about 1 meter away from the breeding holes and loosen the soil. The dimensions of the planting holes are 80cm (length) * 60cm (width) * 15cm (depth). Then, lay a layer of 15cm long substrate at a spacing of about 2cm. Fill the holes with clean, fine soil until 1 / 3 of the substrate surface is exposed. Place the Armillaria mellea evenly at both ends and sides of the substrate. Cover the substrate with clean, fine soil until the substrate is no longer visible. Then, lay another layer of substrate, place the Armillaria mellea on top, cover with soil, and cover with about 10cm of fallen leaves. Step 3: Planting Polygonatum: From October to December of the second year, dig up and select the propagated Gastrodia elata seeds to obtain seeds; then collect the fungal material in the breeding holes and crush it into pieces of about 2-3cm, then spread it evenly in the Gastrodia elata breeding holes, add quicklime to the Gastrodia elata breeding holes, plow to 15cm, cover the surface with the excavated soil, then prepare the land and make ridges. In March of the following year, plant Polygonatum elata at 6 plants / hole, select disease-free tubers as seeds, and after sowing, cover with about 8cm of soil and about 10cm of fallen leaves. Step Four: Planting Gastrodia elata and Polygonatum sibiricum: First, remove the surface covering and soil from the substrate. Take out the substrate from the surface layer, then continue digging down to the bottom layer. Pull out 1 / 3 of the substrate. Place the selected 1 / 2 of the Gastrodia elata seeds evenly at both ends and sides of the substrate. Cover with clean, fine soil until the seeds are no longer visible. Then, arrange the removed substrate at approximately 2cm intervals, fill with clean, fine soil, leaving 1 / 3 of the substrate exposed. Place the remaining seeds evenly at both ends and sides of the substrate, and cover with fine soil until the seeds are no longer visible. Afterwards, cover with 2cm of soil, and finally cover with 10cm of fallen leaves. In November-December of the third year, after harvesting Gastrodia elata, collect the fungal material in the planting hole and crush it into pieces of about 2-3cm. Then, spread it evenly in the Gastrodia elata breeding hole, add quicklime to the Gastrodia elata breeding hole, plow to 15cm, cover the surface with the excavated soil, and then prepare the land and make ridges. In March of the following year, plant Polygonatum sibiricum at a rate of 6 plants / hole, select disease-free tubers as seeds, and after planting, cover with about 8cm of soil and 10cm of fallen leaves. Step 5: Cyclic planting: After two years of planting Polygonatum, harvest the remaining stems, leaves and fine roots and return them to the field. Then, follow the methods in Steps 1 to 4 to lay out new substrate and plant Gastrodia elata seeds.

[0024] Example 2: A method for treating Gastrodia elata in old acupoints repeatedly includes the following steps: Step 1: Propagation of Gastrodia elata: From March to July of the first year, select suitable wasteland for planting Gastrodia elata, dig breeding holes and loosen the soil. The specifications of the breeding holes are 80cm (length) * 60cm (width) * 15cm (depth). Then, lay a layer of 10cm long mycelium material at a spacing of about 2cm. Then fill it with clean fine soil until 1 / 3 of the mycelium material surface is exposed. Place Armillaria mellea and germinated Gastrodia elata seeds evenly at both ends and sides of the mycelium material. Place fresh branch nodes between the Armillaria mellea and germinated Gastrodia elata seeds to promote the branching growth of Armillaria mellea mycelium. Then cover it with clean fine soil until the mycelium material is no longer visible. Then lay a layer of mycelium material, and place the Armillaria mellea and Gastrodia elata seeds. After covering with soil, cover with about 12cm of fallen leaves to help retain heat and moisture.

[0025] Step 2: Cultivating the substrate: After the breeding is completed, dig planting holes about 1 meter away from the breeding holes and loosen the soil. The dimensions of the planting holes are 80cm (length) * 60cm (width) * 15cm (depth). Then, lay a layer of 10cm long substrate at 2cm intervals. Fill the holes with clean, fine soil until 1 / 3 of the substrate surface is exposed. Place the Armillaria mellea evenly at both ends and sides of the substrate. Cover the substrate with clean, fine soil until the substrate is no longer visible. Then, lay another layer of substrate, place the Armillaria mellea on top, cover with soil, and cover with about 10cm of fallen leaves. Step 3: Planting Polygonatum: From October to December of the second year, dig up and select the propagated Gastrodia elata seeds to obtain seeds; then collect the fungal material in the breeding holes and crush it into 2-3cm pieces, then evenly spread it in the Gastrodia elata breeding holes, while adding quicklime to the breeding holes, tilling to 20cm, covering the surface with the excavated soil, then preparing the land and making ridges. In March of the following year, plant Polygonatum elata at 6 plants / hole, selecting disease-free tubers as seeds, and after sowing, cover with about 8cm of soil and about 12cm of fallen leaves. Step Four: Planting Gastrodia elata and Polygonatum sibiricum: First, remove the surface covering and soil from the substrate. Take out the substrate from the surface layer, then continue digging down to the bottom layer. Pull out 1 / 3 of the substrate. Place the selected 1 / 2 of the Gastrodia elata seeds evenly at both ends and sides of the substrate. Cover with clean, fine soil until the seeds are no longer visible. Then, arrange the removed substrate at 2cm intervals, fill with clean, fine soil, leaving 1 / 3 of the substrate exposed. Place the remaining seeds evenly at both ends and sides of the substrate, and cover with fine soil until the seeds are no longer visible. Afterwards, cover with 1cm of soil, and finally cover with 12cm of fallen leaves. In November-December of the third year, after harvesting Gastrodia elata, collect the fungal material in the planting hole and crush it into 2-3cm pieces. Then, spread it evenly in the Gastrodia elata breeding hole, add quicklime to the Gastrodia elata breeding hole, plow to 20cm, cover the surface with the excavated soil, and then prepare the land and make ridges. In March of the following year, plant Polygonatum sibiricum at a rate of 6 plants / hole, select disease-free tubers as seeds, and cover with about 8cm of soil and 12cm of fallen leaves after planting. Step 5: Cyclic planting: After planting Polygonatum for 3 years, harvest it. After harvesting, dig up the remaining stems, leaves and fine roots and turn them back into the field. Then, follow the methods in Steps 1 to 4 to lay out new fungal materials and plant Gastrodia elata seeds.

[0026] Example 3: A method for treating Gastrodia elata in old acupoints repeatedly includes the following steps: Step 1: Propagation of Gastrodia elata: From March to July of the first year, select suitable wasteland for planting Gastrodia elata, dig breeding holes and loosen the soil. The specifications of the breeding holes are 80cm (length) * 60cm (width) * 15cm (depth). Then, lay a layer of 20cm long mycelium material at a spacing of about 3cm. Then fill it with clean fine soil until 1 / 3 of the mycelium material surface is exposed. Place Armillaria mellea and germinated Gastrodia elata seeds evenly at both ends and sides of the mycelium material. Place fresh branch nodes between the Armillaria mellea and germinated Gastrodia elata seeds to promote the branching growth of Armillaria mellea mycelium. Then cover it with clean fine soil until the mycelium material is no longer visible. Then lay a layer of mycelium material, place the Armillaria mellea and Gastrodia elata seeds, and cover with soil and about 8cm of fallen leaves to help retain heat and moisture.

[0027] Step 2: Cultivating the substrate: After the breeding is completed, dig planting holes about 1 meter away from the breeding holes and loosen the soil. The dimensions of the planting holes are 80cm (length) * 60cm (width) * 15cm (depth). Then, lay a layer of 20cm long substrate at 3cm intervals. Fill the holes with clean, fine soil until 1 / 3 of the substrate surface is exposed. Place the Armillaria mellea evenly at both ends and sides of the substrate. Cover the substrate with clean, fine soil until the substrate is no longer visible. Then lay another layer of substrate, place the Armillaria mellea on top, cover with soil, and cover with about 10cm of fallen leaves. Step 3: Planting Polygonatum: From October to December of the second year, dig up and select the propagated Gastrodia elata seeds to obtain seeds; then collect the fungal material in the breeding holes and crush it into 2-3cm pieces, then evenly spread it in the Gastrodia elata breeding holes, while adding quicklime to the breeding holes, tilling to 20cm, covering the surface with the excavated soil, then preparing the land and making ridges. In March of the following year, plant Polygonatum at 4 plants / hole, selecting disease-free tubers as seeds, and after sowing, cover with about 8cm of soil and then cover with about 8-12cm of fallen leaves. Step Four: Planting Gastrodia elata and Polygonatum sibiricum: First, remove the surface covering and soil from the substrate. Take out the substrate from the surface layer, then continue digging down to the bottom layer. Pull out 1 / 3 of the substrate. Evenly place the selected 1 / 2 of the Gastrodia elata seeds from Step Three at both ends and sides of the substrate. Cover with clean, fine soil until the seeds are no longer visible. Then, arrange the removed substrate at 3cm intervals, fill with clean, fine soil, leaving 1 / 3 of the substrate exposed. Evenly place the remaining seeds at both ends and sides of the substrate, and cover with fine soil until the seeds are no longer visible. After planting, cover with 3cm of soil, and finally cover with 8cm of fallen leaves. After harvesting Gastrodia elata in November-December of the third year, collect the fungal material in the planting hole and crush it into 2-3cm pieces. Then, spread it evenly in the Gastrodia elata breeding hole, add quicklime to the Gastrodia elata breeding hole, plow to 20cm, cover the surface with the excavated soil, and then prepare the land and make ridges. In March of the following year, plant Polygonatum sibiricum at a rate of 6 plants / hole, select disease-free tubers as seeds, and cover with about 8cm of soil and 8cm of fallen leaves after planting. Step 5: Cyclic planting: After planting Polygonatum for 4 years, harvest it. After harvesting, dig up the remaining stems, leaves and fine roots and turn them back into the field. Then, follow the methods in Steps 1 to 4 to lay out new fungal materials and plant Gastrodia elata seeds.

[0028] Example 4: A method for treating Gastrodia elata in old acupoints repeatedly includes the following steps: Step 1: Propagation of Gastrodia elata: From March to July of the first year, select suitable wasteland for planting Gastrodia elata, dig breeding holes and loosen the soil. The specifications of the breeding holes are 80cm (length) * 60cm (width) * 15cm (depth). Then, lay a layer of 15cm long mycelium material at a spacing of about 2.5cm. Then fill it with clean fine soil until 1 / 3 of the mycelium material surface is exposed. Place Armillaria mellea and the mixed germinated Gastrodia elata seeds evenly at both ends and sides of the mycelium material. Place fresh branch nodes between the Armillaria mellea and the germinated Gastrodia elata seeds to promote the branching growth of the Armillaria mellea mycelium. Then cover it with clean fine soil until the mycelium material is no longer visible. Then lay a layer of mycelium material, and then place the Armillaria mellea and Gastrodia elata seeds. After covering with soil, cover with about 10cm of fallen leaves to help retain heat and moisture.

[0029] Step 2: Cultivating the substrate bed: After the breeding is completed, dig planting holes about 1 meter away from the breeding holes and loosen the soil. The dimensions of the planting holes are 80cm (length) * 60cm (width) * 15cm (depth). Then, lay a layer of 15cm long substrate at 2.5cm intervals. Fill the holes with clean, fine soil until 1 / 3 of the substrate surface is exposed. Place the Armillaria mellea evenly at both ends and sides of the substrate. Cover the substrate with clean, fine soil until the substrate is no longer visible. Then lay another layer of substrate, place the Armillaria mellea on top, cover with soil, and cover with about 10cm of fallen leaves. Step 3: Planting Polygonatum: From October to December of the second year, dig up and select the propagated Gastrodia elata seeds to obtain seeds; then collect the fungal material in the breeding holes and crush it into 2-3cm pieces, then evenly spread it in the Gastrodia elata breeding holes, add quicklime to the Gastrodia elata breeding holes, plow to 20cm, cover the surface with the excavated soil, then prepare the land and make ridges. In March of the following year, plant Polygonatum elata at 6 plants / hole, select disease-free tubers as seeds, and after sowing, cover with about 8cm of soil and about 10cm of fallen leaves. Step Four: Planting Gastrodia elata and Polygonatum sibiricum: First, remove the surface covering and soil from the substrate. Take out the substrate from the surface layer, then continue digging down to the bottom layer. Pull out 1 / 3 of the substrate. Place the selected 1 / 2 of the Gastrodia elata seeds evenly at both ends and sides of the substrate. Cover with clean, fine soil until the seeds are no longer visible. Then, arrange the removed substrate at intervals of about 2.5cm, fill with clean, fine soil, leaving 1 / 3 of the substrate exposed. Place the remaining seeds evenly at both ends and sides of the substrate, and cover with fine soil until the seeds are no longer visible. After planting, cover with 2cm of soil, and finally cover with 10cm of fallen leaves. After harvesting Gastrodia elata in November-December of the third year, collect the fungal material in the planting hole and crush it into 2-3cm pieces. Then, spread it evenly in the Gastrodia elata breeding hole, add quicklime to the Gastrodia elata breeding hole, plow to 20cm, cover the surface with the excavated soil, and then prepare the land and make ridges. In March of the following year, plant Polygonatum sibiricum at a rate of 6 plants / hole, select disease-free tubers as seeds, and cover with about 8cm of soil and 10cm of fallen leaves after planting. Step 5: Cyclic planting: After 5 years of planting Polygonatum, harvest the remaining stems, leaves and fine roots and return them to the field. Then, follow the methods in Steps 1 to 4 to lay out new substrate and plant Gastrodia elata seeds.

[0030] Experimental Example 1: I. Experimental Design: Three treatments were designed: the rotation treatment (LZ) followed by crop rotation with Polygonatum sibiricum and then replanting Gastrodia elata (as in Example 1); the fallow treatment (XG) followed by two years of fallow followed by replanting Gastrodia elata; and the control treatment (ZC) using surrounding natural soil for planting Gastrodia elata. A randomized block design was used, with three plots, each containing five planting holes, and three replicates. The hole area was 0.5 m². 2 The amount of fungal material used is 20 kg·m³. -2 The amount of hemp seed used is 1.0 kg·m-2 The fallen leaves are fern leaves, and are 8-10cm thick.

[0031] II. Indicator Measurement and Methods: (1) Gastrodia elata yield and morphological indicators: The length and diameter of the Gastrodia elata tuber were measured with vernier calipers, and the weight of the Gastrodia elata was weighed with a 1 / 100 scale. The number of healthy new tubers, diseased new tubers, rotten mother tubers and healthy mother tubers were counted. The grading and output value of Gastrodia elata were based on existing research. The length and thickness of the Armillaria mellea mycelium cords were measured with vernier calipers, and the tensile strength of the mycelium cords was measured with a spring balance. The dry weight of the mycelium cords was determined by the drying method (Armillaria mellea was dried to constant weight in an oven at 80℃). The incidence rate of Gastrodia elata is the percentage of newly planted Gastrodia elata (with black spots and lesions on the surface exceeding one-fifth) out of the total number of newly planted Gastrodia elata. This indicator reflects the susceptibility of newly planted Gastrodia elata after treatment. The mother plant rot rate is the percentage of rotten mother plants (with black spots and rot exceeding one-fifth) in the nest out of the total number of mother plants. This indicator reflects the rottenness of mother plants after treatment. The Armillaria mellea infection index is the percentage of wood nodes infected with Armillaria mellea (more than one-fifth infected) out of the total number of wood nodes. This index reflects the vigorous growth of Armillaria mellea after treatment. The formulas for calculating the Gastrodia elata susceptibility rate, mother plant rot rate, and Armillaria mellea infection index are as follows: Incidence rate of Gastrodia elata = Number of infected Gastrodia elata ÷ Total number of Gastrodia elata surveyed × 100% Hemp seed decay rate = Number of decayed hemp seeds ÷ Total number of hemp seeds surveyed × 100% Armillaria mellea infection index = Number of infected wood nodes ÷ Total number of wood nodes surveyed × 100% (2) Quality Indicators of Gastrodia elata: The quality determination of Gastrodia elata medicinal materials was carried out in accordance with the Chinese Pharmacopoeia (2020 edition). The moisture content was determined according to General Rule 0832 (Method II), the total ash content according to General Rule 2302, and the alcohol-soluble extract according to General Rule 2201. The content of gastrodin and p-hydroxymethyl methanol was determined by high performance liquid chromatography, according to General Rule 0512. The dry weight of Gastrodia elata is the percentage of the corresponding fresh weight of the dried product that meets the moisture requirements of the Chinese Pharmacopoeia (2020 edition), and the formula is as follows: Dry weight percentage = Dry weight ÷ Fresh weight × 100% (3) Soil fertility indexes Soil fertility indexes refer to "Soil Agrochemical Analysis" (3rd edition). Soil pH value is determined by pH meter; organic matter is determined by potassium dichromate volumetric method and external heating method; alkaline nitrogen is determined by alkaline diffusion method; available phosphorus is determined by molybdenum antimony colorimetric method; available potassium is determined by flame photometry.

[0032] (4) Soil microbial testing was commissioned to Shanghai Meiji Biomedical Technology Co., Ltd.

[0033] (5) Soil enzyme activity detection: Polyphenol oxidase, laccase, acid phosphatase, and catalase were detected in the soil of each treatment of continuous cropping of Polygonatum rotation. Enzyme activity was determined using the plant enzyme activity and soil enzyme activity kit from Beijing Solarbio Science & Technology Co., Ltd.

[0034] III. Experimental Results:

[0035] Table 1 shows that the yield of fresh Gastrodia elata under the crop rotation treatment was 1.54 kg / m³. 2 The yield of fresh Gastrodia elata after fallow treatment was 0.93 kg / m³. 2 The crop rotation treatment increased yield by 0.66% compared to the fallow treatment, but the difference was not statistically significant; the fresh Gastrodia elata yield in the continuous cropping treatment was 1.91 kg / m². 2 The yield was not significantly different from the crop rotation treatment, but significantly higher than the fallow treatment. Among the various grades of Gastrodia elata, the yield of Gastrodia elata showed the order of continuous cropping > crop rotation > fallow, with a yield of 1.83 kg / m². 2 1.31 kg / m 2 0.83kg / m 2 The yield of white Gastrodia elata was highest in the rotation treatment, followed by fallow, and lastly in the regular cropping treatment, with no significant differences among the three treatments. In terms of yield at each grade, the fallow treatment had no first-grade Gastrodia elata yield, while the yield of first-grade Gastrodia elata in the regular cropping and rotation treatments was not significantly different. The yield of second-grade Gastrodia elata in both the regular cropping and rotation treatments was significantly higher than that in the fallow treatment. In the third and fourth grades, there were no significant differences between any of the three treatments. Regarding the yield and value of dried Gastrodia elata, the order was regular cropping > rotation > fallow, with regular cropping significantly higher than fallow. The difference between the rotation treatment and the regular cropping and fallow treatments was not significant. Regarding the dry weight reduction rate of Gastrodia elata, there were no significant differences among the three treatments. Regarding the disease incidence rate of new Gastrodia elata, the fallow treatment had a disease incidence rate of 24.98%, significantly higher than the regular cropping and rotation treatments. Regarding the seed rot rate, the mother hemp rot rates in the regular cropping, rotation, and fallow treatments were 41.16%, 52.64%, and 63.19%, respectively, with no significant difference between the rotation treatment and the fallow treatment. In summary, crop rotation with Polygonatum sibiricum is more effective in increasing the yield of Gastrodia elata compared to fallow, and the yield of Gastrodia elata is not significantly different from that of the regular crop. Crop rotation with Polygonatum sibiricum can significantly reduce the disease incidence of new Gastrodia elata, reaching the same level as the regular crop, and also reduces the rate of rot in the mother plant.

[0036] As shown in Table 2, compared with fallow, crop rotation of Polygonatum odoratum increases the content of gastrodin and p-hydroxybenzyl alcohol.

[0037] Table 3 shows that crop rotation with Polygonatum and fallow had no significant effect on the tensile strength and thickness of Armillaria mellea mycelia. The tensile strength and mycelia thickness of both treatments were not significantly different from those of the continuous cropping treatment. In terms of mycelia dry weight, the continuous cropping treatment had the highest mycelia dry weight at 30.76 g / m². 2 The dry weight of the mycelium in the crop rotation treatment was significantly higher than that in the fallow treatment; there was no significant difference between the mycelium dry weight in the crop rotation treatment and the continuous cropping treatment. Regarding the *Armillaria mellea* infection index, the order was continuous cropping > crop rotation > fallow, with no significant difference between the crop rotation treatment and the continuous cropping treatment, while the fallow treatment was significantly lower than the continuous cropping treatment. In conclusion, crop rotation with *Polygonatum sibiricum* is more conducive to the growth of *Armillaria mellea*, increasing its dry weight and infection index.

[0038] As shown in Table 4, the soil pH values ​​for crop rotation and fallow treatments were 7.92 and 8.07, respectively, while the pH value for continuous cropping treatment was 7.36. Both crop rotation and fallow treatments significantly increased soil pH compared to continuous cropping treatment. Both crop rotation and fallow treatments increased soil organic matter content, but the difference was not significant compared to continuous cropping. Regarding available nitrogen content, the available nitrogen content in both fallow and crop rotation treatments was higher than that in continuous cropping treatment, with the fallow treatment showing a significantly higher content. There was no significant difference in available nitrogen content between crop rotation and continuous cropping treatments. Regarding available phosphorus content, the available phosphorus content in the fallow treatment was 18.72 mg / kg, while the available phosphorus contents in the continuous cropping and crop rotation treatments were 5.0 mg / kg and 5.17 mg / kg, respectively. There was no significant difference in available phosphorus content between crop rotation and continuous cropping treatments, but the available phosphorus content in the fallow treatment was significantly higher than that in continuous cropping treatment. Regarding available potassium content, the order was fallow > continuous cropping > crop rotation, with significant differences between each of the three treatments. In summary, crop rotation with Polygonatum sibiricum is more effective than fallow in reducing soil pH; crop rotation with Polygonatum sibiricum can reduce the content of available nitrogen and available phosphorus in the soil to the same level as regular crop rotation.

[0039] Crop rotation treatment has a significant effect on soil enzyme activity in Gastrodia elata planting sites, such as... Figure 1As shown in the figures, the soil polyphenol oxidase activity was highest in the crop rotation treatment (17.38 U / g), significantly higher than the fallow treatment (13.02 U / g), but not significantly different from the continuous cropping treatment (17.00 U / g). For soil laccase activity, the activities in the continuous cropping, crop rotation, and fallow treatments were 94.33 U / g, 49.28 U / g, and 18.99 U / g, respectively, with significant differences between each treatment. For acid phosphatase activity, the crop rotation treatment had the highest activity (20,365.30 U / g), with no significant difference from the continuous cropping and fallow treatments. For catalase activity, the order from strongest to weakest was continuous cropping > crop rotation > fallow, with values ​​of 18.73 U / g, 18.38 U / g, and 17.03 U / g, respectively; there was no significant difference in catalase activity among the three treatments. In summary, crop rotation with Polygonatum significantly increased the activities of soil polyphenol oxidase and laccase compared to fallow, with polyphenol oxidase activity reaching the same level as the continuous cropping treatment. Crop rotation and fallow treatments had no significant effect on acid phosphatase and catalase activities.

[0040] In this experimental example, Venn diagrams were used to analyze the similarity and overlap of species composition in each treatment, such as... Figure 2 The three treatments of continuous cropping, crop rotation, and fallow had 519, 750, and 473 unique out-of-plant (OUT) values, respectively, totaling 803 core OUT values. The results indicate that crop rotation with *Polygonatum sibiricum* can influence the composition of the soil fungal community in *Gastrodia elata* to some extent. Each treatment accumulated a certain number of unique OUT values, with the crop rotation treatment having the largest number of unique OUT values.

[0041] The relative abundance of dominant species in each treatment is as follows: Figure 3As shown in Figure a, species with a relative abundance of less than 1% at the phylum level were grouped into "others". The dominant fungal community composition at the phylum level was the same across treatments, but their relative abundance differed. The distribution of the relative abundance of dominant fungal phyla across treatments was as follows: Ascomycota (47.39%–67.87%), Mucoromycota (12.75%–34.70%), Basidiomycota (5.91%–13.20%), and Rozellomycota (0.83%–2.22%). Ascomycota and Mucoromycota had relative abundances above 80% in all treatments, making them the dominant phyla in all three treatments. Ascomycota had the highest relative abundance in the crop rotation treatment, significantly higher than in the continuous cropping and highest abundance treatments. The relative abundance of Mucoromycota was significantly higher in the crop rotation and fallow treatments than in the regular cropping treatment.

[0042] The trend of relative abundance at the level of soil fungi is as follows: Figure 3As shown in b, the relative richness of the top 50 fungal genera reached over 50%. Fungal community composition analysis indicated that the dominant species at the genus level were the same across the three treatments, but their relative richness varied significantly. The relative richness distribution of the dominant genera was as follows: *Mortierella* (12.37%–34.20%), *Trichoderma* (2.93%–12.34%), *Clonostachys* (1.11%–10.64%), and so on. The genera *Volutella* (0.66%–5.16%), *Tomentella* (0.12%–6.15%), *Fusarium* (1.43%–1.47%), *Cylindrocarpon* (0.53%–1.78%), *Cladosporium* (0.16%–1.51%), and *Ilyonectria* (0.11%–0.65%) were abundant. The *Mortierella* genus had the highest relative abundance in the crop rotation and fallow treatments, significantly higher than in the crop rotation and fallow treatments. *Trichoderma* had the highest relative abundance in the crop rotation treatment, significantly higher than in the crop rotation and fallow treatments. In the fallow treatment, *Clonostachys* had the highest relative abundance, significantly higher than in the crop rotation and crop rotation treatments. Common pathogens include *Fusarium*, *Cladosporium*, and *IIyonectria*. The relative abundance of *Fusarium* in the crop rotation, crop rotation, and fallow treatments was 1.47%, 1.45%, and 1.43%, respectively. The relative abundance of *Cladosporium* in the crop rotation, crop rotation, and fallow treatments was 0.85%, 0.53%, and 1.78%, respectively. *IIyonectria* had the highest relative abundance in the fallow treatment at 0.65%, while the relative abundance in the crop rotation and crop rotation treatments was 0.11% and 0.25%, respectively, significantly lower than that in the fallow treatment.

[0043] Depend on Figure 4 It can be seen that the variances of the principal component 1 (PC1) and principal component 2 (PC2) related to soil fungal communities were 38.41% and 30.86% respectively, with a cumulative contribution rate of 69.27%, indicating that there were differences in community composition among the three treatments at the genus level. Figure 4The confidence region distribution of the three treatments shows that the confidence regions of the crop rotation treatment and the regular cropping treatment are clustered together, indicating that the genus-level fungal community composition of the crop rotation treatment is similar. The confidence regions of the fallow treatment and the regular cropping treatment are separated, and the two regions are separated by the principal component (PC2), indicating that the genus-level fungal community composition of the fallow treatment is different from that of the regular cropping treatment.

Claims

1. A method for growing Gastrodia elata in old Gastrodia elata planting holes, characterized in that, Includes the following steps: Step 1: Propagation of Gastrodia elata: From March to July of the first year, select suitable wasteland for planting Gastrodia elata, dig breeding holes and loosen the soil. Then, lay a layer of fungal material with a spacing of ≥2cm. Fill the hole with clean fine soil until 1 / 3 of the fungal material surface is exposed. Place Armillaria mellea and germinated Gastrodia elata seeds evenly at both ends and sides of the fungal material. Cover with clean fine soil until the fungal material is no longer visible. Then lay another layer of fungal material, place Armillaria mellea and Gastrodia elata seeds, cover with soil and fallen leaves. Step 2: Cultivating the substrate: After completing Step 1, dig planting holes and loosen the soil at a distance from the breeding holes. Then, lay a layer of substrate at a spacing of ≥2cm. Fill the holes with clean, fine soil until 1 / 3 of the substrate surface is exposed. Place the Armillaria mellea evenly at both ends and sides of the substrate. Cover the substrate with clean, fine soil until the substrate is no longer visible. Then, lay another layer of substrate, place the Armillaria mellea, cover with soil, and cover with fallen leaves. Step 3: Planting Polygonatum: From October to December of the second year, dig up the propagated Gastrodia elata seeds and select them to obtain Gastrodia elata seeds; then collect the fungal material in the breeding holes and screen out the incompletely decomposed parts, crush them to obtain fragments, and spread them evenly in the Gastrodia elata breeding holes. At the same time, add quicklime to the Gastrodia elata breeding holes, turn the soil, cover the surface with the excavated soil, then prepare the land and make ridges. Sow Polygonatum in March of the year following the harvest, using disease-free tubers as seeds. After sowing, cover with soil and cover with fallen leaves or straw from the forest. Step Four: Planting Gastrodia elata and Polygonatum sibiricum: Selected Gastrodia elata seeds are inoculated onto a fungal bed. Gastrodia elata is harvested in November-December of the third year. The fungal material from the planting holes is then collected, and any incompletely decomposed parts are pulverized and evenly spread into the Gastrodia elata planting holes. Simultaneously, quicklime is added to the planting holes, the soil is turned over, and the surface is covered with excavated soil. The land is then prepared by ridging. Polygonatum sibiricum is planted in March of the following year. After sowing, the seeds are covered with soil, then with fallen leaves or straw from the forest floor. Step 5: Cyclic planting: After planting Polygonatum for 2-5 years, harvest it. After harvesting, turn over the remaining stems, leaves and fine roots on the spot and return them to the field. Then, follow the methods of Step 1 to Step 4 to lay out new fungal material and plant Gastrodia elata seeds.

2. A method of growing Gastrodia elata in old Gastrodia elata planting holes according to claim 1, wherein, In step one, fresh branches and wood nodes are placed between the Armillaria mellea and the germinated Gastrodia elata seeds to promote the branching growth of the Armillaria mellea mycelium.

3. The method for continuous cultivation of Gastrodia elata at old acupoints as described in claim 1, characterized in that, The thickness of the fallen leaves is 8-12 cm.

4. The method for continuous cultivation of Gastrodia elata at old acupoints as described in claim 1, characterized in that, The Polygonatum seedlings used for planting are selected from disease-free tubers that are 2 years old or older.

5. The method for continuous cultivation of Gastrodia elata at old acupoints as described in claim 1, characterized in that, In step four, the inoculation includes the following steps: 1) First, remove the covering material and soil from the surface of the fungal substrate, take out the fungal substrate placed on the surface, and then continue to dig down to the bottom layer of fungal substrate. Pull out 1 / 3 of the substrate surface, and evenly place 1 / 2 of the seed spawn at both ends and sides of the fungal substrate. Then cover it with clean fine soil until the seed spawn is no longer visible. 2) Arrange the removed fungal materials at intervals of ≥2cm, fill them with clean fine soil, leaving 1 / 3 of the material exposed, place the remaining seeds evenly at both ends and sides of the fungal material, cover them with fine soil until the seeds are no longer visible, cover with soil 1-3cm, and finally cover with fallen leaves.

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