Lyophyllum cavaleriei strain L6894 and cultivation method thereof

Abstract

The invention relates to a lyophyllum trabeum strain L6894 and a cultivation method thereof, and belongs to the technical field of microorganisms, the lyophyllum trabeum strain L6894 is preserved in Guangdong Microbial Culture Collection Center on December 17, 2025, and the preservation number is GDMCC NO: 67497. Compared with the currently cultivated lyophyllum decastes, the lyophyllum decastes strain L6894 has larger sporocarp, pileus is grey orange or orange, the pileus is hemispherical when young, and the pileus is gradually flattened after maturing; the stipe is cylindrical, thick and strong, slightly thicker towards the base part, light grey yellow or grey orange, smooth in surface, white and solid in mushroom pulp, free of color change after being injured, and wide in suitable temperature range.

Description

TECHNICAL FIELD

[0001] The application belongs to the technical field of microorganisms, and particularly relates to a Lyophyllum connatum strain L6894 and a cultivation method thereof. BACKGROUND

[0003] Lyophyllum connatum in Yunnan Province is mainly sold in the form of wild fruiting bodies, and artificial planting is rarely successful. Lyophyllum connatum has poor resistance to other bacteria, a long growth cycle, high requirements for fruiting conditions, low yield and technical difficulties, and is difficult to be large-scale cultivated and popularized. Although there are many domestication and cultivation research reports, the mycelium culture conditions differ greatly due to differences in regions and research methods, and further verification and exploration are needed. SUMMARY

[0004] In order to overcome the problems in the background art, the application provides a Lyophyllum connatum strain L6894 and a cultivation method thereof. The Lyophyllum connatum strain L6894 has been preserved in the Guangdong Microbial Culture Collection Center on December 17, 2025, with a preservation number of GDMCC No: 67497, and the address of the preservation unit is the 5th floor of Building 59, Guangdong Academy of Sciences, 100 Xianlie Road, Guangzhou.

[0005] The application also provides application of the Lyophyllum connatum strain L6894 in Lyophyllum connatum breeding.

[0006] The application also provides a cultivation method of the Lyophyllum connatum strain L689, which comprises the following steps:

[0007] (1) The mycelium of the Lyophyllum connatum strain L6894 is used for propagation by using a modified PDA culture medium, and is cultured at 24 DEG C in the dark, and the mother strain is obtained after the culture medium is fully covered;

[0008] (2) The mother strain cultured in step (1) is transferred to a liquid culture medium, and is cultured in the dark in a constant-temperature shaking bed to obtain a liquid strain;

[0009] (3) The liquid strain of step (2) is inoculated into a cultivation bag, and is cultured in the dark in a constant-temperature incubator at 24 DEG C until the mycelium fully covers the cultivation bag;

[0010] (4) The cultivation bag cultured in step (3) is transported to a greenhouse for soil cultivation, and the fruiting bodies are harvested when the fruiting bodies grow to maturity and the caps are flat.

[0011] Further, in step (1), the modified PDA culture medium is composed of 200±10 g of potatoes, 1 L of water, 20±2 g of malt powder, 2±0.1 g of soybean powder, 0.5±0.1 g of calcium carbonate, 0.002±0.0002 g of inositol, and 20±3 g of agar powder.

[0012] Furthermore, in step (2), the liquid culture medium is composed of the following components: 200±10g of potato, 1 L of water, 20±2g of malt extract powder, 2±0.1g of soybean powder, 0.5±0.1g of calcium carbonate, and 0.002±0.0002g of inositol.

[0013] Furthermore, in step (3), the cultivation material in the cultivation bag includes the following components by weight: 75-80 parts sawdust, 15-20 parts wheat bran, 1-2 parts sugar, 1-2 parts gypsum, and 1-2 parts quicklime.

[0014] The beneficial effects of this invention are:

[0015] This invention obtained strain L6894 of Lyophyllum aurantiacum by culturing and domesticating Lyophyllum aurantiacum collected from Beixin Village, Mouding County, Chuxiong Prefecture, Yunnan Province. Phylogenetic analysis of strain L6894 was performed using the ITS and LSU gene sequences. The maximum likelihood method was used to calculate the branch node support rate, which was 97%. The strain was identified as Lyophyllum aurantiacum, thus determining its taxonomic position.

[0016] The *Amanita fulva* strain L6894 of this invention exhibits rapid mycelial growth, with thick, white mycelia. After cultivation, the cap size is (5.80±0.50) × (4.20±0.40) cm, and the fresh weight is 12.47±7.76 g / cap. Each package can yield 2-3 harvests. Compared to the currently cultivated *Amanita fulva*, the *Amanita fulva* var. *lotus* has a larger fruiting body, with a grayish-orange or orange cap, a cylindrical, pale grayish-yellow or grayish-orange stipe, a smooth surface, and a thicker stipe. The flesh is white and solid, does not change color when injured, and the cap is darker in color.

[0017] Compared to wild fruiting bodies, the fruiting bodies of the *Agaricus cirrhosa* strain L6894 of this invention are more uniform (fruiting occurs annually in October-November with soil covering), and the fruiting bodies are larger (800-1200 g per cluster). The overall fruiting is uniform, and the fruiting time of soil covering cultivation is mainly in autumn and winter, which is different from the peak season of wild mushrooms (July-September), forming an off-season advantage and a wide suitable temperature range. Attached Figure Description

[0018] Figure 1 These are images of the three wild strains collected in this invention after domestication;

[0019] Figure 2 This invention is a maximum likelihood (ML) phylogenetic tree of the genus *Pyroxylon* constructed based on ITS + LSU multigenes;

[0020] Figure 3 A comparison of the effects of different carbon sources on the mycelial growth of *Agaricus cirrhosa* strain;

[0021] Figure 4 A comparison of the effects of different nitrogen sources on the mycelial growth of *Agaricus cirrhosa* strain;

[0022] Figure 5 A comparison of the effects of different inorganic salts on the mycelial growth of *Agaricus cirrhosa* strains;

[0023] Figure 6 A comparison of the effects of different growth factors on the mycelial growth of *Agaricus cirrhosa* strain;

[0024] Figure 7 A comparison of the effects of different temperatures on the mycelial growth of *Agaricus cirrhosa* strain;

[0025] Figure 8 This is a diagram illustrating the cultivation process of the orange-capped pleated cap mushroom according to the present invention; ac represents the primordia stage, df represents the young mushroom stage, and gi represents the maturity stage. Detailed Implementation

[0026] To make the objectives, technical solutions, and beneficial effects of this invention clearer, the technical solutions of this invention will be described in detail below. Obviously, the described embodiments are only a part of the embodiments of this invention, and not all of them. Other embodiments obtained by those skilled in the art based on the embodiments of this invention without creative effort are all within the scope of protection of this invention.

[0027] Example 1: Obtaining and identifying *Amanita fulva* strain L6894

[0028] (1) Collection and domestication

[0029] Wild fruiting bodies of *Gnaphalium affine* from Beixin Village, Mouding County, Chuxiong Prefecture: Cap 1.8-9 cm wide, fleshy and crisp, hemispherical when young, gradually flattening as mature, with a dry and smooth surface; gills 0.4-0.8 cm wide, grayish-yellow, not discolored when injured, with intact edges; stipe cylindrical, 4.5-12.5 × 0.5-2.1 cm, slightly thickening towards the base, pale grayish-yellow or grayish-orange, with a smooth surface and white, solid flesh.

[0030] One *Amanita muscaria* plant was collected, and three different parts of the best-performing wild fruiting body were isolated using tissue isolation: cap flesh (G), cap-stipe junction flesh (GB), and stipe flesh (B). These parts were inoculated onto PDA medium to obtain three strains for domestication. The three isolated strains were then incubated at 22°C for screening. After 15 days of incubation, the growth of the three strains was as follows: Figure 1 As shown, strain GB (with the densest mycelium and fastest growth rate) was selected. Figure 1 The strain on the far right was named *Amanita fulva* strain L6894.

[0031] (2) Preparation of culture medium

[0032] Modified PDA medium: 200 g potato, 20 g malt extract powder, 2 g soybean flour, 0.5 g calcium carbonate, 0.002 g inositol, 20 g agar powder, 1 L water.

[0033] (3) Strain domestication

[0034] The obtained strain L6894 was propagated using modified PDA medium and cultured at 18 ± 1℃ in the dark.

[0035] (4) Molecular biology research

[0036] Genome extraction: Hyphae of *Amanita fulva* strain L6894 were added to a 1.5 mL centrifuge tube with zircon beads. The tube was then placed in a rapid biological sample preparation system for low-temperature disruption and centrifugation. DNA was extracted using the Ezup column fungal genomic DNA extraction kit from Sangon Biotech (Shanghai) Co., Ltd., and the extracted DNA was stored at -20 ℃. Two gene fragments, ITS and LSU, were selected. The specific primer base composition is shown in Table 1. RNA polymerase chain reaction (PCR) was used to amplify the ITS and LSU gene fragments. The PCR reaction system (25 μL) included: 12.5 μL mix, 9.5 μL sterile ddH2O, 1 μL each of forward and reverse primers at a concentration of 100 mol / μL, and 1 μL of stock DNA solution. The primers used for amplification of the two gene fragments are shown in the table below.

[0037] Table 1 Primers and base composition used for PCR amplification

[0038]

[0039] Note: Bases include A, T, C, and G. Other letters in the primers represent degenerate bases.

[0040] The PCR amplification reaction program was as follows: initial denaturation at 95 ℃ for 3 min, 35 cycles (denaturation at 94 ℃ for 30 s, denaturation at 53 ℃ for 30 s, denaturation at 72 ℃ for 1 min), and extension at 72 ℃ for 10 min.

[0041] Detection of PCR amplification products: Dissolve 0.8 g of agarose in 80 mL of 0.5×TBE solution and heat in a microwave oven for 1 min to accelerate dissolution until clear and transparent. After slightly cooling, add 8 μL of nucleic acid dye to the solution, shake well, transfer to a silica gel plate, insert a comb, and allow it to cool before use. Place the cooled agarose gel in an electrophoresis tank, add electrophoresis buffer (0.5×TBE solution), and use a pipette to spot the marker and PCR amplification products at a volume of 2 μL. Then, electrophoresis is performed for 15 min at 110 V and 80 A. After electrophoresis is completed and the power is turned off, the agarose gel is removed and observed in a UV gel imaging system. If the bands are clear, the PCR amplification is successful. The PCR products are sent to Sangon Biotech (Shanghai) Co., Ltd. for sequencing to obtain the characteristic sequences of strain L6894: ITS (as shown in SEQ ID NO.1) and LSU (as shown in SEQ ID NO.2).

[0042] Sequence results were searched using Blast in the GenBank database on the NCBI website and compared with existing 5.8S and 28S ribosomal RNA genes. The obtained strain L6894 sequence showed high similarity to *Lyophyllum*. Sequences of 5.8S and 28S ribosomal RNA genes with high homology to strain L6894 were downloaded and aligned using MAFFT v.7 (https: / / mafft.cbrc.jp / alignment / server / ). Each gene fragment was further examined in BioEdit v.7. Maximum likelihood (ML) analysis was performed on the CIPRES platform using RAxML-HPC2 v.8.2.3, employing the GTR+G+I model, and a phylogenetic tree was constructed. The results are as follows: Figure 2 As shown, the branches are marked with nodes where the maximum likelihood support is equal to or greater than 75% and the Bayesian posterior probability is equal to or greater than 0.90. Lyophyllum aurantiacum is marked in bold red. The results show that strain L6894 and Lyophyllum aurantiacum HKAS 147010 clustered together. The sequences were checked and compared, and the difference was 2 bases. Therefore, strain L6894 was identified as Lyophyllum aurantiacum.

[0043] Bayesian analysis was performed in MrBayes v.3.2, and the optimal alternative model for each partition was determined by MrModeltest v.2.3: the GTR+G model was used for ITS and LSU. The Markov Chain Monte Carlo (MCMC) method was used to estimate the posterior probability (PP). Six chains were run for 50,000 generations, with the stoprul parameter set to stopval = 0.01, and sampling was performed every 100 generations. A branch was considered to have strong support if its bootstrap support (BS) ≥ 70% and posterior probability (PP) ≥ 0.90. Developmental tree analysis showed that the orange-capped Lyophyllum aurantiacum strain L6894 clustered with another orange-capped Lyophyllum specimen, exhibiting high bootstrap support; strain L6894 was therefore identified as orange-capped Lyophyllum aurantiacum.

[0044] After successfully domesticated and cultivated fruiting bodies of Lyophyllum aurantiacum L6894 were isolated and preserved for biological collection at the Guangdong Provincial Microbial Culture Collection Center. Taxonomic name: Lyophyllum aurantiacum, accession number GDMCC No: 67497, address of the collection institution is Institute of Microbiology, Guangdong Academy of Sciences, 5th Floor, Building 59, No. 100 Xianlie Middle Road, Guangzhou.

[0045] Example 2: Effect of carbon source on the mycelial growth of *Amanita fulva* L6894

[0046] Carbon sources tested: glucose, malt extract, sucrose, fructose, and lactose.

[0047] The basic culture medium formula used was: 1 L water, 2 g peptone, 0.5 g magnesium sulfate, and 20 g agar.

[0048] One of the tested carbon sources was added to the basal culture medium at a concentration of 20 g / L (20 g carbon source per liter of culture medium). A control group without carbon source was used. After mixing, the mixture was poured into 250 mL Erlenmeyer flasks (200 mL per flask) and sterilized at 121 °C for 2 h. After slight cooling, the mixture was poured into plates, approximately 30 mL per plate. A 7 mm diameter Lyophyllum aurantiacum L6894 fruiting body was inoculated in the center of each plate (the same applies to the following examples). Each treatment was repeated 6 times. The plates were incubated at 22 °C. Colony diameter was measured every 3 days using the cross-sectional method. Measurements were stopped when the mycelium completely covered the culture medium. Mycelial growth rate, density, and color were recorded and photographed. The mycelial growth under different carbon sources was obtained (Table 2 and...). Figure 3The growth rate showed a pattern of malt extract powder > fructose > sucrose > glucose > lactose, indicating that the mycelial growth rate of strain L6894 was the fastest when malt extract powder was used as the carbon source, which was (3.67±0.52) mm / d.

[0049] Table 2. Effects of carbon source on the mycelial growth of *Amanita fulva* L6894.

[0050]

[0051] Note: Mycelial density and lateral growth can be described using "+", where "+++" indicates dense mycelial growth / good lateral growth, "++" indicates relatively dense mycelial growth / average lateral growth, and "+" indicates sparse mycelial growth / poor lateral growth; different lowercase letters indicate significant differences (p<0.05); the rest are described in detail.

[0052] Example 3: Effect of nitrogen source on the mycelial growth of *Pleurotus ostreatus* L6894

[0053] Nitrogen sources tested: peptone, soybean flour, urea, yeast powder, and beef extract.

[0054] The basic culture medium formula used is: 1 L water, 20 g glucose, 0.5 g magnesium sulfate, and 20 g agar.

[0055] One of the tested nitrogen sources was added to the basal culture medium at a concentration of 2 g / L (2 g nitrogen source per liter of culture medium). A control group without nitrogen source was used. After mixing, the mixture was poured into 250 mL Erlenmeyer flasks (200 mL per flask) and sterilized at 121 °C for 2 h. After slight cooling, the mixture was poured into plates, approximately 30 mL per plate. A 7 mm diameter mycelial block was inoculated in the center of each plate. Each treatment was replicated six times. The plates were incubated at 22 °C. Colony diameter was measured every 3 days using the cross-crossing method. Measurements were stopped when mycelia completely covered the culture medium. Mycelial growth rate, density, and color were recorded and photographed. The mycelial growth under different nitrogen sources was obtained (Table 3 and...). Figure 4 The results showed a pattern of soybean flour > peptone > yeast powder > urea > beef extract, indicating that the mycelial growth rate of strain L6894 was the fastest (4.33±0.00) mm / d when soybean flour was used as the nitrogen source.

[0056] Table 3. Effects of nitrogen source on the mycelial growth rate of *Amanita fulva* L6894.

[0057]

[0058] Note: Mycelial density and lateral growth can be described using "+", where "+++" indicates dense mycelial growth / good lateral growth, "++" indicates relatively dense mycelial growth / average lateral growth, and "+" indicates sparse mycelial growth / poor lateral growth; different lowercase letters indicate significant differences (p<0.05); the rest are described in detail.

[0059] Example 4: Effects of Inorganic Salts on the Mycelial Growth of *Pleurotus ostreatus* L6894

[0060] The inorganic salts tested were: magnesium sulfate, potassium dihydrogen phosphate, ferrous sulfate, calcium chloride, and calcium carbonate.

[0061] The basic culture medium formula used is: 1 L water, 20 g glucose, 2 g peptone, and 20 g agar.

[0062] One of the tested inorganic salts was added to the basal culture medium at a concentration of 0.5 g / L (0.5 g of inorganic salt per liter of culture medium). A control group without inorganic salt was used. After mixing, the solutions were poured into 250 mL Erlenmeyer flasks (200 mL per flask) and sterilized at 121 °C for 2 h. After slight cooling, the solutions were poured onto plates, approximately 30 mL of culture medium per plate. A 7 mm diameter mycelial block was inoculated in the center of each plate. Each treatment was replicated six times. The plates were incubated at 22 °C. Colony diameter was measured every 3 days using the cross-crossing method. Measurements were stopped when mycelia completely covered the culture medium. Mycelial growth rate, density, and color were recorded and photographed. The mycelial growth under different inorganic salt conditions was obtained (Table 4 and...). Figure 5 The results showed a pattern of calcium carbonate > calcium chloride > magnesium sulfate > potassium dihydrogen phosphate > ferrous sulfate, indicating that when calcium carbonate was chosen as the inorganic salt, the mycelial growth rate of strain L6894 was the fastest (3.36±0.11) mm / d.

[0063] Table 4. Effects of inorganic salts on the mycelial growth of *Amanita fulva* L6894.

[0064]

[0065] Note: Mycelial density and lateral growth can be described using "+", where "+++" indicates dense mycelial growth / good lateral growth, "++" indicates relatively dense mycelial growth / average lateral growth, and "+" indicates sparse mycelial growth / poor lateral growth; different lowercase letters indicate significant differences (p<0.05); the rest are described in detail.

[0066] Example 5: Effects of growth factors on the mycelial growth of *Pleurotus ostreatus* L6894.

[0067] Tested growth factors: Vitamin B1, Vitamin B6, Vitamin C, Vitamin E, and Inositol

[0068] The basic culture medium formula used is: 1 L water, 20 g glucose, 2 g peptone, 0.5 g magnesium sulfate, and 20 g agar.

[0069] One of the tested growth factors was added to the basal medium at a rate of 0.002 g. A control group without growth factor was used. After mixing, the mixture was poured into 250 mL Erlenmeyer flasks (200 mL per flask) and sterilized at 121 °C for 2 h. After slight cooling, the mixture was poured into plates, approximately 30 mL per plate. A 7 mm diameter mycelial block was inoculated in the center of each plate. Each treatment was replicated six times. The plates were incubated at 22 °C for 30 days. Colony diameter was measured every 3 days using the cross-hatching method. Measurements were stopped when the culture medium was completely covered. Mycelial growth rate, density, and color were recorded and photographed. The mycelial growth under different growth factors was obtained (Table 5 and...). Figure 6 The results showed that inositol > vitamin B1 > vitamin C > vitamin E > vitamin B6, indicating that the mycelial growth rate of strain L6894 was the fastest (2.86±0.02) mm / d when inositol was used as the growth factor.

[0070] Table 5. Effects of growth factors on the mycelial growth of *Amanita fulva* L6894.

[0071]

[0072] Note: Mycelial density and lateral growth can be described using "+", where "+++" indicates dense mycelial growth / good lateral growth, "++" indicates relatively dense mycelial growth / average lateral growth, and "+" indicates sparse mycelial growth / poor lateral growth; different lowercase letters indicate significant differences (p<0.05); the rest are described in detail.

[0073] Example 6: Effect of temperature on the mycelial growth of *Pleurotus ostreatus* L6894

[0074] Test temperatures: 18 ℃, 20 ℃, 22 ℃, 24 ℃, 26 ℃, 28 ℃.

[0075] The basic culture medium formula used is: 1 L water, 20 g glucose, 2 g peptone, 0.5 g magnesium sulfate, and 20 g agar.

[0076] Prepare the basal medium by dispensing 200 mL of culture medium into 250 mL Erlenmeyer flasks. Sterilize each flask at 121 °C for 2 hours. After slight cooling, pour the medium onto plates, approximately 30 mL per plate. Inoculate the center of each plate with a 7 mm diameter mycelial block. Each treatment is replicated six times. Incubate at constant temperatures of 18 °C, 20 °C, 22 °C, 24 °C, 26 °C, and 28 °C for 30 consecutive days. Measure the colony diameter every 3 days using the cross-hatching method. Stop measurement once the culture medium is fully covered. Record the mycelial growth rate, density, and color, and take photographs. Obtain the mycelial growth at different temperatures (Table 6 and...). Figure 7 The mycelial growth rate was highest at 22 ℃ > 24 ℃ > 26 ℃ > 20 ℃ > 28 ℃ > 18 ℃, with the fastest growth rate (3.23±0.03) mm / d at 22 ℃. The mycelial growth rate showed very little difference between 20 and 26 ℃, indicating that the strain has a wide suitable temperature range.

[0077] Table 6. Effects of temperature on the mycelial growth of *Amanita fulva* L6894.

[0078]

[0079] Note: Mycelial density and lateral growth can be described using "+", where "+++" indicates dense mycelial growth / good lateral growth, "++" indicates relatively dense mycelial growth / average lateral growth, and "+" indicates sparse mycelial growth / poor lateral growth; different lowercase letters indicate significant differences (p<0.05); the rest are described in detail.

[0080] Example 7 Cultivation of Orange-Lidded Umbrella L6894

[0081] (1) Activation, rejuvenation and propagation of strains

[0082] After activating the mycelium of *Pleurotus ostreatus* L6894 at room temperature, it was inoculated onto modified PDA medium and placed in an incubator at 24 ℃ for constant temperature incubation until the colonies filled the culture dish.

[0083] The modified PDA culture medium consists of: 200g potato, 1L water, 20g malt extract, 2g soybean flour, 0.5g calcium carbonate, 0.002g inositol, and 20g agar powder.

[0084] (2) Preparation of liquid bacterial strains

[0085] Take the mother culture prepared in step (1) and transfer it to a liquid culture medium shake flask. Incubate in a constant temperature shaker at 26 ℃ and 170 r / min in the dark. When the number of bacterial balls in the liquid shake flask accounts for two-thirds of the total liquid volume, the liquid culture is obtained.

[0086] Liquid culture medium: 200 g potato, 1 L water, 20 g malt extract powder, 2 g soybean flour, 0.5 g calcium carbonate, 0.002 g inositol.

[0087] (3) Preparation of cultivars

[0088] The liquid inoculum from step (2) is inoculated into a cultivation bag and placed in a 24 ℃ incubation room for dark culture, maintaining an air humidity of 60-65%. Once the mycelium has fully grown in the cultivation bag, the culture is obtained.

[0089] The cultivation material in the cultivation bag includes the following components by weight: 75-80 parts sawdust, 15-20 parts wheat bran, 1-2 parts sugar, 1-2 parts gypsum, and 1-2 parts slaked lime.

[0090] (4) Soil covering cultivation

[0091] Transport the spawn from step (3) to the greenhouse, and dig planting troughs 0.80-1.00 m wide and 0.10-0.20 m deep. Carefully remove the plastic from the mushroom bags with a knife, being careful not to damage the mycelium. Place the removed cultivation bags horizontally in the planting troughs, keeping a spacing of 5-7 cm between the cultivation bags and a row spacing of 10-15 cm. Cover the planting troughs with soil, with a thickness of 2-3 cm, and then carry out mushroom management.

[0092] (5) Results of domestication and cultivation

[0093] The mycelium of *Pleurotus ostreatus* L6894 is pure white, dense, and grows slowly. About 65 days after inoculation, the mycelium fills the bag. After removing the bag, the bag is covered with soil for cultivation. Primordia begin to appear after (20±5) days, young mushrooms begin to appear after (30±5) days, and mature after (40±5) days.

[0094] Domesticated and cultivated fruiting bodies are relatively large, with a grayish-orange center and a pale orange edge that is rolled inwards; the flesh is white and does not change color after injury; the stipe is cylindrical, slightly swollen towards the base, and is pale grayish-yellow or grayish-orange; the stipe flesh is white and solid, and does not change color after injury. See the growth status diagram of orange-capped *Gymnocladus orientalis* L6894 during cultivation. Figure 8The cap size is (5.80±0.50)×(4.20±0.40) cm, the thickness is (0.30±0.10) cm, the stipe size is (9.50±0.80)×(1.30±0.20) cm, the fresh weight of each mushroom is (12.47±7.76) g, and there are 8 to 14 mushrooms per cluster. Compared with wild fruiting bodies, the number of mushrooms per cluster is significantly increased, and the mushroom weight is increased by 50% to 80%. The fruiting rate of the bag is 66.7%, the biological efficiency is 48.42%, and it can produce 2 to 3 flushes of fruiting. During the domestication and cultivation process, mushrooms can be produced from June to October. It can still grow well under conditions of 18-26 ℃ and 75% humidity, with a wider suitable temperature range than the currently cultivated *Pleurotus ostreatus*. The fruiting bodies of the currently cultivated *Pleurotus ostreatus* are smaller. For example, the cap diameter of *Pleurotus ostreatus* is 1-3 cm, with a smoky color, and the stipe is about 5-7 cm long and 1-3 cm thick. After domestication and cultivation, the cap diameter of the *Pleurotus ostreatus* fruiting bodies is 5-6 cm, which is about 175% larger than that of the currently cultivated *Pleurotus ostreatus*. The stipe size is 8.5-9.5 cm × 1.0-1.5 cm, which is about 55% longer than that of the currently cultivated *Pleurotus ostreatus*, resulting in a better commercial shape.

[0095] The basic principles, core features, and technical advantages of this invention have been fully disclosed above. Those skilled in the art will understand that the scope of protection of this invention is not limited to the described embodiments; the embodiments and the content contained in the specification are only for illustrative purposes. Any changes, optimizations, or improvements made to this invention without departing from its spirit and without exceeding the scope defined by its claims should be covered within the scope of protection of this invention. The scope of the claims of this invention is determined by the appended claims and their equivalents.

Claims

1. A strain of *Amanita fulva* L6894, characterized in that, It was deposited at the Guangdong Provincial Center for Microbial Culture Collection on December 17, 2025, with accession number GDMCC NO:67497.

2. The application of the *Amanita fulva* strain L6894 according to claim 1 in the breeding of *Amanita fulva*.

3. A method for cultivating *Amanita fulva* strain, characterized in that, The *Amanita fulva* strain L6894 described in claim 1 was cultivated.

4. The cultivation method according to claim 3, characterized in that, Includes the following steps: (1) Using modified PDA medium, the mycelium of *Agaricus cirrhosa* strain L6894 was propagated and cultured at 24 ℃ in the dark until the medium was fully grown to obtain the mother culture. (2) Take the mother culture cultured in step (1) and transfer it to liquid culture medium. Place it in a constant temperature shaker and culture it in the dark to obtain liquid culture. (3) Inoculate the liquid culture from step (2) into the cultivation bag and place it in a 24 ℃ constant temperature incubator to cultivate in the dark until the mycelium has covered the cultivation bag; (4) Transport the cultivation bags prepared in step (3) to the greenhouse for soil covering and cultivation. Harvest the fruiting bodies when they have grown to maturity and the caps are flat.

5. The cultivation method according to claim 4, characterized in that, In step (1), the modified PDA culture medium consists of 200±10 g of potato, 1 L of water, 20±2 g of malt extract powder, 2±0.1 g of soybean powder, 0.5±0.1 g of calcium carbonate, 0.002±0.0002 g of inositol, and 20±3 g of agar powder.

6. The cultivation method according to claim 4, characterized in that, In step (2), the liquid culture medium consists of the following components: 200±10g potato, 1 L water, 20±2g malt extract powder, 2±0.1g soybean powder, 0.5±0.1g calcium carbonate, and 0.002±0.0002g inositol.

7. The cultivation method according to claim 4, characterized in that, In step (3), the cultivation material in the cultivation bag includes the following components by weight: 75-80 parts sawdust, 15-20 parts wheat bran, 1-2 parts sugar, 1-2 parts gypsum, and 1-2 parts quicklime.

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