Boletus pumilum PP013 and its application
By using spore inoculation of Boletus pumilus strain PP013 and dry weed mulching technology, the problems of unstable mycelial colonization and unstable yield in existing cultivation methods have been solved, achieving efficient and sustainable understory fruiting and ecological cycle.
Patent Information
- Application Number
- CN202610473217.7
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2026-04-10
- Publication Date
- 2026-05-26
AI Technical Summary
Existing methods for cultivating Boletus edulis under forest cover suffer from problems such as unstable mycelial colonization success rate, high cost, short duration of fruiting effect, and unstable yield, making it impossible to form a stable ecological cycle.
Using the Boletus truncatus strain PP013, spores from its fruiting bodies were inoculated. Combined with shallow-rooted trees and a layer of dry weeds, a self-sustaining, cyclical fruiting symbiotic system was established. Through steps such as liquid spawn cultivation, spawn cultivation, soil covering for fruiting, and understory inoculation, a sustainable fruiting model was formed.
This achieved stable mycelial colonization and efficient growth, reduced production costs, increased mushroom yield, formed a sustainable ecological cycle, and promoted biodiversity in the forest ecosystem.
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Abstract
Description
Technical Field
[0001] This invention relates to the field of microbiology, specifically to a Boletus pueraria strain PP013 and its applications. Background Technology
[0002] Boletus edulis is a valuable edible fungus, generally considered to be a facultative parasite. Existing understory cultivation techniques usually involve directly inoculating artificially cultured strains (usually mycelium) into the forest. This method has several drawbacks: the success rate of mycelium colonization in complex forest environments is unstable, the inoculation amount is large, the cost is high, the fruiting effect is short-lived, the yield is unstable, and repeated inoculation is often required for many years, making it impossible to form a stable ecological cycle.
[0003] Therefore, developing a cultivation method that utilizes its natural reproduction process under artificial intervention is crucial for the industrialization of wild Boletus thunbergii. Summary of the Invention
[0004] The purpose of this invention is to overcome the shortcomings of existing technologies and provide a *Boletus pustulosa* strain PP013 and its applications. This invention can be used for sustainable, cyclical fruiting cultivation in forests. The core of this method is not simply transplanting mycelium, but rather selecting the most suitable shallow-rooted tree species, introducing artificially cultivated fruiting bodies as "spore bombs" into a suitable forest environment, utilizing the large number of basidiospores produced for inoculation, and establishing a self-sustaining, cyclical fruiting symbiotic system under artificial intervention.
[0005] To achieve the above objectives, the technical solution designed by the present invention is as follows: This invention provides a dark brown reticulate Boletus ( Phlebopus portentosus )PP013, with accession number CGMCC No.40661.
[0006] The above-mentioned dark brown reticulate Boletus ( Phlebopus portentosus PP013 was deposited on June 6, 2023, at the China General Microbiological Culture Collection Center (CGMCC) with accession number CGMCC No. 40661, located at No. 3, Courtyard 1, Beichen West Road, Chaoyang District, Beijing.
[0007] The present invention also provides the application of the above-mentioned Boletus PP013 in the preparation of protoplasts, spores, fruiting bodies or mycelia.
[0008] The present invention also provides an application of the above-mentioned Boletus PP013 in the cultivation of edible fungi or under-forest cultivation.
[0009] This invention also provides a sustainable, cyclical mushroom cultivation method under forest cover, comprising the following steps: (1) Liquid culture: The above strains were inoculated into liquid culture medium and cultured in the dark with shaking to obtain liquid culture; (2) Cultivation of culture medium: Inoculate the liquid culture medium into the sterilized culture medium and cultivate in the dark until the mycelium fills the bag; (3) Covering the mushroom bags with soil: Cover the surface of the cultured bags with soil, control the temperature, humidity and light, and cultivate until the fruiting bodies are mature; (4) Understory inoculation and sustainable fruiting cycle: Select shallow-rooted tree species, remove the top layer of soil 0.8-1.5 meters away from the center of the trunk, lay a layer of dry weeds, place the fruiting bodies with gills about to open along with the mushroom bags under the tree to produce spores, lay a layer of dry weeds again after the spores are produced, water the soil and the covering layer to make them thoroughly wet, and after the fruiting of the following year, each tree should retain 2-3 fruiting bodies for spore production each time, allowing them to mature and produce spores naturally, water in time during the dry season, and replenish the layer of dry weeds.
[0010] Furthermore, in step (4), the shallow-rooted tree species are one or more of the following: crape myrtle, Bauhinia purpurea, gardenia, flame tree, and coffee.
[0011] Furthermore, the shallow-rooted tree species is either the flame tree or the gardenia.
[0012] Furthermore, in step (4), the dry weeds include sugarcane bagasse, miscanthus and barnyardgrass, and the dry weight ratio of the three is 1:1:1; The thickness of the topsoil layer removed is 1-5cm, the thickness of the first layer of dry weeds is 1-5cm, and the thickness of the dry weeds layer laid after spore production is 3-5cm.
[0013] Furthermore, in step (1), the liquid culture medium is P1 liquid culture medium, which consists of: 200 g / L peeled potato, 20 g / L glucose, 4 g / L yeast extract, 2 g / L MgSO4, and 2 g / L K2HPO4.
[0014] Furthermore, in step (2), the culture medium for the cultivation of the seedlings consists of: 12 kg of rubberwood chips, coffee wood chips or miscellaneous wood chips, 7.5 kg of sugarcane bagasse, 24 kg of mountain soil, 12 kg of grains, 6 kg of flour, 15 g of MgSO4, 15 g of KH2PO4, pH value 5.0-6.0, and water added to make the water content of the culture medium 50%-60.0%.
[0015] Furthermore, in step (3), the thickness of the soil cover layer is 3-5cm, the temperature is 25-30℃, the relative humidity is 80-95%, and the illuminance is 400-1500Lx.
[0016] Furthermore, in step (4), the shallow-rooted tree species are 3-5 years old.
[0017] The beneficial effects of this invention are: This invention breaks away from the traditional thinking of direct mycelial inoculation and creatively uses artificial living fruiting body spores as strains, returning to and utilizing the fungus's natural reproductive instinct, making the technical approach more ecological and efficient.
[0018] By covering with dry weeds and intervening with artificial watering, a stable, high-humidity microenvironment is provided for spore germination. Direct sunlight is avoided to prevent damage to spores and young mycelia. A loose and breathable structure is formed, which is conducive to mycelial colonization and growth. Furthermore, by planting once and retaining some fruiting bodies, a self-reproducing mycelial community is established, achieving benefits for many years with a single inoculation and greatly reducing long-term production costs.
[0019] This method is carried out entirely under the forest canopy, without damaging the main structure of the forest land. It is a highly efficient forest-fungus complex model that opens up new avenues for increasing the income of forest farmers and promotes the biodiversity of forest ecosystems. Attached Figure Description
[0020] Figure 1 The colony morphology of Boletus PP013 strain on different culture media is shown. In the figure, A shows the colony morphology of Boletus pumilum PP013 strain on P1 solid medium; B shows the colony morphology of Boletus PP013 strain on PDA medium. C shows the colony morphology of Boletus PP013 strain on Delonix regia medium. D is a colony morphology diagram of Boletus PP013 strain on sugarcane bagasse medium. E shows the colony morphology of Boletus PP013 strain on reed culture medium. F shows the colony morphology of Boletus PP013 strain on Miscanthus agar medium. G is a colony morphology diagram of Boletus PP013 strain on Digitaria rubra culture medium. H is a colony morphology diagram of Boletus PP013 strain on Cimicifuga foetida medium. I is a colony morphology diagram of Boletus PP013 strain on guar gum medium; J shows the colony morphology of Boletus PP013 strain on rubberwood culture medium.
[0021] Figure 2 Figure showing the effect of different culture media on the mycelial growth of Boletus pueraria var. PP013. In the diagram, A represents P1 solid medium; B represents PDA medium; C represents Delonix regia medium; D represents sugarcane bagasse medium; E represents reed medium; F represents Miscanthus sinensis medium; G represents Crataegus pinnatifida medium; H represents Crataegus pinnatifida medium; I represents Guillaume medium; and J represents Rubberwood medium. Figure 3 Image of liquid culture of Boletus PP013 (dark brown reticulate stalk). Figure 4 This is a diagram of indoor soil covering and fruiting of Boletus PP013 strain (dark brown reticulate stem). Figure 5 This image shows the fruiting of Boletus PP013 strain under a Delonix regia forest. Figure 6 This image shows the fruiting of Boletus PP013 strain under Gardenia jasminoides trees. Figure 7 This image shows the fruiting of Boletus PP013 strain under a Crape Myrtle forest. Figure 8 This image shows the fruiting of Boletus PP013 strain in a coffee forest. Figure 9 This image shows the fruiting of Bauhinia variegata strain PP013 under forest cover. Detailed Implementation The present invention will now be described in further detail with reference to specific embodiments, so that those skilled in the art can understand it.
[0022] Example 1: Isolation and Identification of Boletus purpureus PP013 1. Materials Strain PP013 was collected from wild fruiting bodies in Nongdao Town, Ruili City, Dehong Prefecture, Yunnan Province on September 10, 2020, and obtained through isolation and purification.
[0023] 2. Isolation and culture of bacterial strains Fresh, robust, and disease-free wild Boletus edulis fruiting bodies were collected. In a clean bench, the tissue at the junction of the cap and stipe was surface-sterilized with 75% alcohol swabs. Approximately 5 mm³ internal tissue blocks were excised using a sterile scalpel and inoculated into the center of sterilized P1 medium plates (200 g / L peeled potato, 20 g / L glucose, 4 g / L yeast extract, 2 g / L MgSO4, 2 g / L K2HPO4, 15 g / L agar). The plates were incubated at 25-28℃ in the dark. After mycelial growth, the tips of the mycelia were selected for purification to obtain a pure culture strain, named PP013, which was then preserved.
[0024] 3. Morphological identification The isolated and purified strain PP013 was inoculated onto the above-mentioned P1 medium and placed at (25-28)℃ for dark culture. After 14 days, mycelia were picked and slides were prepared for observation of mycelial morphology under an optical microscope. Colony morphology characteristics were observed and measured daily.
[0025] 4. DNA sequencing and molecular identification DNA was extracted from the strain using the Tsingke DNA extraction kit. The extracted DNA sample was diluted appropriately and used as a PCR template. Gene primers were used to amplify the strain's genomic DNA via PCR. The ITS primer pair was as follows: ITS1: 5′-TCCGTAGGTGAACCTGCGG-3′, as shown in SEQ ID NO:2; ITS4: 5′-TCCTCCGCTTATTGATATGC-3′, as shown in SEQ ID NO:3; The amplification system consisted of 1 μL of primers, 1 μL of template DNA, and 45 μL of Tsingke TSE101 Gold Mix. PCR amplification was performed on an amplification instrument using the following program: 94℃ pre-denaturation for 2 min, 94℃ denaturation for 10 s, 56℃ annealing for 10 s, 72℃ extension at a rate of 10 s / kb, 35 cycles, followed by a 5 min extension at 72℃, with the final annealing temperature at 56℃. The amplified PCR products were analyzed by agarose gel electrophoresis (2 μL sample + 6 μL bromophenol blue) and observed and photographed using a gel imaging system. The prepared PCR products were sent to Kunming Qingke Biotechnology Co., Ltd. for sequencing; the nucleotide sequence of the ITS gene is shown in SEQ ID NO:1. ; The measured sequences are submitted to the NCBI database, and Blast is used to perform homology comparisons with known sequences.
[0026] 5. Results After obtaining strain PP013, its morphological characteristics were identified. ITS-rDNA sequence analysis of strain PP013 was performed, and the sequence results were compared with those in the NCBI database using the BLAST program. Phlebopus portentosus(Accession number PV029065.1) The highest homology was 99.74%. Combining morphological characteristics and relevant molecular biological information, strain PP013 is *Boletus glomeratus* (accession number PV029065.1). Phlebopus portentosus It was named Boletus purpurea PP013.
[0027] The above-mentioned dark brown reticulate Boletus ( Phlebopus portentosus PP013 was deposited on June 6, 2023, at the China General Microbiological Culture Collection Center (CGMCC) with accession number CGMCC No. 40661, located at No. 3, Courtyard 1, Beichen West Road, Chaoyang District, Beijing.
[0028] Example 2: Growth of Boletus purpureus PP013 on different culture media 1. Different culture medium formulations Potato glucose agar (PDA): 200g peeled potatoes, 20g glucose, 15g agar, add distilled water to a final volume of 1000mL.
[0029] P1 solid medium (A): 200g peeled potato, 20g glucose, 4g / L yeast extract, 2g MgSO4, 2g K2HPO4, 15g agar, add distilled water to a final volume of 1000mL.
[0030] PDA medium (B): 200g peeled potato, 20g glucose, 15g agar, add distilled water to a final volume of 1000mL.
[0031] Flame Tree Culture Medium (C): 20g of dried flame tree powder, 15g of agar, and distilled water to a final volume of 1000mL.
[0032] Sugarcane bagasse culture medium (D): 20g sugarcane bagasse powder, 15g agar, add distilled water to a final volume of 1000mL.
[0033] Reed culture medium (E): 20g of dried reed powder, 15g of agar, and distilled water to a final volume of 1000mL.
[0034] Miscanthus culture medium (F): 20g dried Miscanthus powder, 15g agar, add distilled water to a final volume of 1000mL.
[0035] Crab Culture Medium (G): 20g of dried crab powder, 15g of agar, and distilled water to a final volume of 1000mL.
[0036] Cultivated yam culture medium (H): 20g of dried cultivated yam powder, 15g of agar, and distilled water to a final volume of 1000mL.
[0037] Sesbania culture medium (I): 20g of dried sesbania powder, 15g of agar, and distilled water to a final volume of 1000mL.
[0038] Rubberwood culture medium (J): 20g of dry rubberwood powder, 15g of agar, and distilled water to a final volume of 1000mL.
[0039] The aforementioned phoenix wood powder and rubberwood powder are made by chopping and drying phoenix wood and rubberwood branches, then grinding them into powder using a pulverizer. The aforementioned crabgrass, sesbania, guarana, reed, and miscanthus powder are made by chopping and drying the above-ground parts of crabgrass, sesbania, guarana, reed, and miscanthus, then grinding them into powder using a pulverizer. The aforementioned bagasse powder is made from bagasse remaining after sugarcane processing, which is then dried and ground into powder using a pulverizer. The aforementioned culture medium is placed in a 121°C autoclave for 15-20 minutes. After the culture medium cools to approximately 50-60°C, in a sterile operating area near an alcohol lamp flame, the culture medium is poured into pre-sterilized petri dishes (approximately 20 mL / dish).
[0040] 2. Growth on different culture media Standard mycelial discs were prepared from the edge of the pre-cultured PP013 colonies using a 5mm diameter punch. The mycelial discs were inoculated in the center of each treatment medium plate, and the plates were sealed with sterile sealing film. The treatment information was marked with an oil-based marker. All plates were placed in an inverted dark incubator at 28±1℃. Each treatment was replicated in 3 places. After 14 days of incubation, the colony diameter was measured using the cross-sectional method.
[0041] like Figure 1 As shown, the mycelial growth of Boletus PP013 on 10 different culture media was different. The mycelium grew vigorously and densely on P1 solid medium (A) and PDA medium (B), while the growth was weak and sparse on the other media. The colonies were larger on P1 solid medium (A), PDA medium (B) and sugarcane bagasse medium (D).
[0042] Depend on Figure 2 It can be seen that *Boletus pumilum* PP013 can grow on all 10 culture media. The mycelial growth rate is fastest on sugarcane bagasse medium (D), with a colony diameter of 34.63 mm after 14 days of culture, which is significantly larger than that on the other 9 culture media. The next fastest growth rates are on P1 solid medium (A), PDA medium (B), Miscanthus sinensis medium (F), and Dioscorea opposita medium (H). Therefore, sugarcane bagasse, Miscanthus sinensis, and Dioscorea opposita are conducive to the mycelial growth of *Boletus pumilum* PP013.
[0043] Example 3: Fruiting and understory cultivation of Boletus PP013 (dark brown reticulate stem). 1. Liquid culture In a clean bench, the Boletus pueraria lobata PP013 spawn was cut into small pieces. These pieces were inoculated into 500 mL Erlenmeyer flasks containing 200 mL of P1 liquid medium (composition: peeled potato: 200 g / L, glucose: 20 g / L, yeast extract: 4 g / L, MgSO4: 2 g / L, K2HPO4: 2 g / L). The Erlenmeyer flasks were then placed in a shaker at 120-150 r / min and incubated in the dark at 28℃. After 7 days of incubation, the liquid spawn was obtained. Figure 3 ).
[0044] 2. Cultivation of Cultivated Varieties Culture medium for cultivation: 12 kg of rubber, coffee or other hardwood chips, 7.5 kg of sugarcane bagasse, 24 kg of hillside soil, 12 kg of grains, 6 kg of flour, 15 g of MgSO4, 15 g of KH2PO4, pH 5.0-6.0. Add water and mix until the moisture content of the culture medium is 50%-60.0%.
[0045] Wood chips, sugarcane bagasse, and grains are soaked in water to pre-moisten them, then drained and set aside. The prepared spawn culture medium is mixed thoroughly and placed into polypropylene spawn bags, sterilized at 121℃ for 30 minutes, and then cooled before use. The prepared liquid spawn is inoculated into the sterilized spawn culture medium. After inoculation, the culture bags are placed in a 28℃ incubation room in the dark for 30 days until the mycelium has fully colonized the bags.
[0046] 3. Covering with soil to promote mushroom growth For mushroom cultivation with casing soil, sift out larger clods of soil from the base of the plant and then cover the surface of the cultivated mushroom bags with a casing soil layer 3-5 cm thick. Place the bag in a mushroom house at a temperature of 28℃, a relative humidity of 80-95%, and a light intensity of 400-1500 Lx. Keep the surface of the casing soil moist. Primordia will appear after about 7 days, and the fruiting bodies will mature in about 14 days. When the fruiting body cap is fully thickened, darkens in color, is not yet fully flat, and the gills are about to open (…), the mushrooms will mature. Figure 4 Stop watering and prepare for transplanting under the trees.
[0047] 4. Inoculation under forest cover and sustainable fruiting cycle After sugarcane is processed into sugar, the remaining sugarcane bagasse is dried and set aside for later use. The above-ground parts of Miscanthus sinensis and Dioscorea opposita are cut, dried, and set aside for later use.
[0048] Select shallow-rooted tree species such as crape myrtle, bauhinia, gardenia, flame tree, and coffee, which are 3-5 years old. One meter from the center of the trunk, remove 1-5cm of topsoil and cover with 1-5cm of dry weeds (sugarcane bagasse, miscanthus, and crabgrass in a 1:1:1 dry weight ratio). Place the fruiting bodies (with gills about to open) along with the substrate bag (…). Figure 4Place the bags under the trees to await spore production, 1-2 bags per tree. After spore production in 1-2 days, cover with a 3-5cm layer of dry weeds (sugarcane bagasse, miscanthus, and barnyardgrass in a 1:1:1 dry weight ratio); then water thoroughly to saturate the soil and mulch. Mushrooms will appear the following year. After fruiting, retain 2-3 fruiting bodies per tree for spore production each time. Allow unharvested fruiting bodies to mature and produce spores naturally, thus naturally reinforcing the inoculation of the forest and achieving sustainable fruiting cycles. Water promptly during the dry season (to keep the soil moist) and supplement with dry weed mulch to promote fruiting. Figures 5-9 ).
[0049] Comparative Example 1 1. Liquid culture In a clean bench, the Boletus pueraria lobata PP013 spawn was cut into small pieces. These pieces were inoculated into 500 mL Erlenmeyer flasks containing 200 mL of P1 liquid medium (composition: peeled potato: 200 g / L, glucose: 20 g / L, yeast extract: 4 g / L, MgSO4: 2 g / L, K2HPO4: 2 g / L). The Erlenmeyer flasks were then placed in a shaker at 120-150 r / min and incubated in the dark at 28℃. After 7 days of incubation, the liquid spawn was obtained. Figure 3 ).
[0050] 2. Cultivation of Cultivated Varieties Culture medium for cultivation: 12 kg of rubber, coffee or other hardwood chips, 7.5 kg of sugarcane bagasse, 24 kg of hillside soil, 12 kg of grains, 6 kg of flour, 15 g of MgSO4, 15 g of KH2PO4, pH 5.0-6.0. Add water and mix until the moisture content of the culture medium is 50%-60.0%.
[0051] Wood chips, sugarcane bagasse, and grains are soaked in water to pre-moisten them, then drained and set aside. The prepared spawn culture medium is mixed thoroughly and placed into polypropylene spawn bags, sterilized at 121℃ for 30 minutes, and then cooled before use. The prepared liquid spawn is inoculated into the sterilized spawn culture medium. After inoculation, the culture bags are placed in a 28℃ incubation room in the dark for 30 days until the mycelium has fully colonized the bags.
[0052] 3. Inoculation under forest cover Dig up the soil around the roots of 3-5 year old crape myrtle, bauhinia, gardenia, flame tree, and coffee trees to expose the lateral roots. Then break the cultivated spawn into clumps and inoculate them onto the lateral roots, using 500-1000g of spawn per tree. When inoculating, try to make close contact between the spawn and the lateral roots of the tree. Then cover the dug-up soil back up.
[0053] 4. Cultivating mushrooms After inoculation, water thoroughly to keep the soil around the roots of the inoculated tree moist, and mushrooms will grow the following year.
[0054] Table 1. Understory cultivation and fruiting of Boletus PP013 (Dark Brown Net-Stemmed) The results are shown in Table 1: All five shallow-rooted tree species successfully formed a symbiotic relationship with Boletus purpureus PP013, but the host tree species had a significant impact on yield. For example, Delonix regia performed best (386.86 kg / mu under conventional methods, 800.40 kg / mu under optimized methods), followed by Gardenia jasminoides, both of which had significantly higher yields than the other three. Bauhinia purpurea had the lowest yield, only about 62% of that of Delonix regia.
[0055] This invention optimizes management by providing a stable, high-humidity microenvironment for spore germination through timely watering and the laying of dry straw during the dry season. This avoids direct sunlight damage to spores and young mycelia, creating a loose and breathable structure that promotes mycelial colonization and growth. The addition of dry straw (sugarcane bagasse, miscanthus, and crabgrass in a 1:1:1 dry weight ratio) also provides the necessary nutrients for the growth of *Boletus edulis*. Through timely watering and the addition of dry straw during the dry season, the yield of all five tree species under optimized management was approximately 2.07 times that of conventional management, with a highly consistent increase (approximately 107%–109%). This demonstrates that these two measures—watering during the dry season and adding dry straw—are universally effective for all tree species and are key to steadily increasing yield. This invention can double mushroom production through reasonable water management (watering during the dry season) and the covering with dry straw, allowing for 8-10 harvests per year.
[0056] All other parts not described in detail are existing technologies. Although the above embodiments have provided a detailed description of the present invention, they are only some embodiments of the present invention, not all embodiments. People can obtain other embodiments based on these embodiments without creative effort, and these embodiments all fall within the protection scope of the present invention.
Claims
1. A dark brown, net-stalked Boletus ( Phlebopus portentosus )PP013, with accession number CGMCCNo.40661.
2. The use of the Boletus PP013 of claim 1 in the preparation of protoplasts, spores, fruiting bodies or mycelia.
3. The application of the Boletus PP013 of claim 1 in the cultivation of edible fungi or under-forest cultivation.
4. A sustainable, cyclical mushroom cultivation method under forest cover, characterized by: Includes the following steps: (1) Liquid culture: The Boletus PP013 of claim 1 was inoculated into a liquid culture medium and cultured under shaking in the dark to obtain a liquid culture; (2) Cultivation of culture medium: Inoculate the liquid culture medium into the sterilized culture medium and cultivate in the dark until the mycelium fills the bag; (3) Covering the mushroom bags with soil: Cover the surface of the cultured bags with soil, control the temperature, humidity and light, and cultivate until the fruiting bodies are mature; (4) Understory inoculation and sustainable fruiting cycle: Select shallow-rooted tree species, remove the top layer of soil 0.8-1.5 meters away from the center of the trunk, lay a layer of dry weeds, place the fruiting bodies with gills about to open along with the mushroom bags under the tree to produce spores, lay a layer of dry weeds again after the spores are produced, water the soil and the covering layer to make them thoroughly wet, and after the fruiting of the following year, each tree should retain 2-3 fruiting bodies for spore production each time, allowing them to mature and produce spores naturally, water in time during the dry season, and replenish the layer of dry weeds.
5. The forest understory cultivation method according to claim 4, characterized in that: In step (4), the shallow-rooted tree species are one or more of the following: crape myrtle, Bauhinia purpurea, gardenia, flame tree, and coffee.
6. The forest understory cultivation method according to claim 4 or 5, characterized in that: The shallow-rooted tree species are either flame trees or gardenias.
7. The forest understory cultivation method according to claim 4, characterized in that: In step (4), the dry weeds include sugarcane bagasse, Miscanthus sinensis and Dioscorea opposita, and the dry weight ratio of the three is 1:1:1; the thickness of the removed topsoil layer is 1-5cm, the thickness of the first layer of dry weeds is 1-5cm, and the thickness of the layer of dry weeds laid after spore production is 3-5cm.
8. The forest understory cultivation method according to claim 4, characterized in that: In step (1), the liquid culture medium is P1 liquid culture medium, which consists of: 200g / L peeled potato, 20g / L glucose, 4g / L yeast extract, 2g / L MgSO4, and 2g / L K2HPO4. In step (2), the culture medium consists of: 12kg rubberwood chips, coffee wood chips, or miscellaneous wood chips, 7.5kg sugarcane bagasse, 24kg hillside soil, 12kg grains, 6kg flour, 15g MgSO4, 15g KH2PO4, pH 5.0-6.0, and water added to make the water content of the culture medium 50%-60.0%.
9. The forest understory cultivation method according to claim 4, characterized in that: In step (3), the thickness of the soil cover layer is 3-5cm, the temperature is 25-30℃, the relative humidity is 80-95%, and the illuminance is 400-1500Lx.
10. The forest understory cultivation method according to claim 6, characterized in that: In step (4), the shallow-rooted tree species are 3-5 years old.