A strain of thin-skinned casein and its cultivation method and application

Abstract

The present invention provides a Tyromyces chioneus strain, which is Tyromyces chioneus V11 and has a deposit number of GDMCC No. 65077. The present invention also provides a cultivation method for the Tyromyces chioneus, comprising the steps of pre-treatment of culture medium, bagging and sterilization, inoculation and cultivation, post-ripening culture, bud-inducing and fruiting, and fruiting management, to obtain Tyromyces chioneus fruiting bodies, followed by harvesting and post-harvest management, and secondary fruiting. The *Cephalotaxus fortunei* strain of this invention can produce two flushes of mushrooms, with a fruiting body yield of 196.80±5.21 g / bag, a biological efficiency of 35.14±0.93%, a total flavonoid content of 708 mg / 100g, a crude protein content of 16.30 g / 100g, a dietary fiber content of 18.82 g / 100g, and a calcium content of 554 mg / kg. The strain exhibits a lignin degradation rate of 33.93%, a cellulose degradation rate of 37.72%, and a hemicellulose degradation rate of 28.96%.

Description

Technical Field

[0001] This invention belongs to the field of microbial technology, specifically relating to a strain of *Cephalomyces thunbergii* and its cultivation method and application. Background Technology

[0002] Macrofungi germplasm resources are fundamental for cultivating high-yield and high-quality edible fungi varieties, providing abundant materials for research in edible fungi genetics and evolutionary biology. Fully developing and utilizing macrofungi resources to obtain superior germplasm is crucial. However, many macrofungi mycelia often exhibit slow growth, difficulty in preservation, unstable growth cycles, immature propagation techniques, and high domestication difficulty. These factors severely restrict the richness of germplasm resources and delay subsequent research. In a sense, the preservation and domestication cultivation of new macrofungi strains are fundamental and preliminary work. *Tyromyceschioneus*, belonging to the phylum Basidiomycota, class Agaricales, order Polyporaceae, family Polyporaceae, and genus *Tyromyceschioneus*, is widely distributed in temperate and cold-temperate broad-leaved forests in Asia, Europe, and North America. It grows on fallen logs and secretes non-specific extracellular enzyme systems such as laccase and lignin peroxidase, degrading lignocellulose and playing a vital role in promoting the transformation of materials in forest ecosystems. Thin-skinned casein also possesses antioxidant and antiviral biological activities, and studies have found that a variety of compounds can be isolated from the liquid fermentation broth of thin-skinned casein.

[0003] Currently, there is relatively little research on the domestication and cultivation of *Ceratophyllum demersum*. Qi Zhengxiang et al. conducted research on the biological characteristics and domestication of *Ceratophyllum demersum* in *Acta Mycotae Sinica*, 2023, 42(1):408-417. They disclosed that they successfully cultivated fruiting bodies of *Ceratophyllum demersum* on a culture medium containing 77% broadleaf tree sawdust, 20% wheat bran, 1% glucose, 0.8% lime, 0.8% gypsum, 0.3% potassium dihydrogen phosphate, and 0.1% magnesium sulfate. However, the biological efficiency was only 19.15%, and not every bag produced fruiting bodies. The biological efficiency and fruiting rate were both low, and there is an urgent need for high-quality strains to expand the germplasm resource bank of *Ceratophyllum demersum*. Summary of the Invention

[0004] The technical problem to be solved by this invention is to address the shortcomings of the prior art by providing a Tyromyces chioneus strain and its cultivation method and application. The strain is Tyromyces chioneus V11. The cultivation method of this invention can produce two flushes of mushrooms with high fruiting body yield and high biological efficiency. This invention also evaluates the nutritional components of Tyromyces chioneus, providing materials and basis for the widespread cultivation of Tyromyces chioneus and providing a scientific basis for the development and sustainable utilization of macrofungi germplasm resources.

[0005] To solve the above-mentioned technical problems, the technical solution adopted by the present invention is as follows:

[0006] A strain of Tyromyces chioneus, namely Tyromyces chioneus V11, was deposited on August 30, 2024, at the Guangdong Provincial Center for Microbial Culture Collection, with accession number GDMCC NO.65077. The address of the depository is 5th Floor, Building 59, No. 100 Xianlie Middle Road, Guangzhou.

[0007] The present invention also provides a method for cultivating the thin-skinned casei, the method comprising the following steps:

[0008] S1, Culture medium pretreatment

[0009] The cultivation substrate formula by weight percentage is: 40% corn cob, 43% sawdust, 15% wheat bran, 1% gypsum, and 1% lime. Weigh the raw materials according to the proportion, pre-moisten the corn cob and sawdust with clean water 24 hours in advance, and then mix them evenly with wheat bran, gypsum, and lime to obtain the culture medium.

[0010] S2, Packing and Sterilization

[0011] The culture medium obtained in S1 is bagged, and the bagged culture medium is sterilized at 117℃ for 10 hours, then left to sit for 2 hours. When the temperature drops below 70℃, the culture medium is removed from the sterilizer and cooled to below 25℃ before inoculation.

[0012] S3, Inoculation and Culture

[0013] Inoculate with thin-skinned casein under aseptic conditions, seal the container, and then place it in a colony in the dark for incubation. The substrate temperature is controlled at 22℃-25℃. After 10 days, the substrate temperature is adjusted to 20℃-22℃, and the relative humidity is controlled below 40%. Ventilate 1-2 times a day for 0.5 hours each time.

[0014] S4, Post-ripening culture

[0015] After the mycelium of the thin-skinned case fungus has fully colonized the spawn bag, continue to cultivate for 5-10 days, and adjust the indoor temperature to 18℃-22℃.

[0016] S5, Inducing Bud and Mushroom Growth

[0017] Move the mushroom bags into the fruiting room and place them closely together on the ground, along the wall. Stack the bags tightly together, in rows, with 4-6 layers. Leave the bag openings and bottoms slightly apart between layers to create two fruiting surfaces. Use a sterilized blade to make a 10-20mm long "I" shaped cut at the bag opening. After opening the bags, allow the mycelium to recover for 2-3 days. Lower the temperature inside the greenhouse to 18-22℃, maintain the relative humidity at 80%-85%, provide ventilation, and increase diffused light.

[0018] S6, Mushroom Production Management

[0019] After the mushroom buds form, the relative humidity inside the shed should be maintained at 80%-90%, the temperature at 22℃-25℃, the air inside the shed should be kept fresh, and the light intensity should be controlled at 100Lx-400Lx.

[0020] S7 Harvesting

[0021] Harvest once the mushrooms are mature. Stop watering 24 hours before harvesting. Keep your hands and tools clean and hygienic when harvesting. Gently remove the fruiting bodies of the thin-skinned cheese mushroom from the base, without attaching any substrate.

[0022] S8, Post-harvest Management

[0023] After each harvest, promptly clean up any remaining mushrooms and remove any contaminated bags. After harvesting, stop watering for 3-5 days to allow the mycelium to resume growth before proceeding with secondary mushroom cultivation management.

[0024] Preferably, the moisture content of the culture medium in S1 is 55%-60%.

[0025] Preferably, the inoculation amount of the thin-skinned casei in S3 is 8% of the weight of the cultivation substrate.

[0026] The present invention also provides the application of the aforementioned thin-skinned casei in the degradation of matrix lignin, cellulose and hemicellulose.

[0027] The present invention has the following significant technical effects:

[0028] 1. This invention provides a *Tyromyces chioneus* strain V11 and its cultivation method. Using this method, the strain can produce two flushes of mushrooms, with a fruiting body yield of 196.80±5.21 g / bag, a biological efficiency of 35.14±0.93%, a total flavonoid content of 708 mg / 100g, a crude protein content of 16.30 g / 100g, a dietary fiber content of 18.82 g / 100g, and a calcium content of 554 mg / kg. This invention provides materials and a basis for the widespread cultivation of *Tyromyces chioneus* and a scientific foundation for the development and sustainable utilization of macrofungi germplasm resources.

[0029] 2. The Tyromyces chioneus strain V11 of the present invention can be used to degrade lignin, cellulose and hemicellulose. The test results show that the degradation rate of matrix lignin is 33.93%, the degradation rate of cellulose is 37.72% and the degradation rate of hemicellulose is 28.96%.

[0030] The present invention will be further described in detail below with reference to the accompanying drawings and embodiments. Attached Figure Description

[0031] Figure 1 This is a diagram of the primordium of the thin-skinned casein bacteria from Example 2;

[0032] Figure 2 These are images of the fruiting bodies of the thin-skinned cheese fungus from Example 2. Left: Front view; Right: Back view. Detailed Implementation

[0033] Example 1

[0034] This example demonstrates the identification of the bacterial strain.

[0035] The fungal strain was collected in 2019 from the Sandaoguan National Forest Park in Mudanjiang City. Tissue samples were taken from the junction of the cap and stipe, and then placed on PDA medium and incubated in the dark at 25°C for 2 weeks to obtain mycelia. The mycelia were purified twice to obtain fungal strain V11. The purification steps for each step were as follows: the mycelia were placed on PDA medium and incubated in the dark at 25°C for 1 week.

[0036] 1. Morphological identification

[0037] Morphological characteristics of the fruiting body: The fruiting body grows on the fallen branches of broad-leaved trees. It is fleshy, with a fan-shaped cap that extends 6cm outward, is 8cm wide, and is up to 2cm thick at the base. The surface is milky yellow with a slightly thinner edge. The surface of the pores is creamy yellow. The fruiting body is round, with 4-5 pores per millimeter. The edge is thin and entire. The flesh is milky white and up to 2cm thick. The tubes are pale yellowish-brown.

[0038] 2. Molecular biological identification (ITS)

[0039] (1) Genomic DNA of fungus V11 was extracted and used as a template. It was then amplified by PCR using primers ITS1 and ITS4 to obtain PCR amplification products. The nucleotide sequence of primer ITS1 is shown in SEQ ID NO.1, and the nucleotide sequence of primer ITS4 is shown in SEQ ID NO.2.

[0040] (2) Take the PCR amplification product obtained in step (1) and sequence it.

[0041] The nucleotide sequence of the product is shown in SEQ ID NO.3. The nucleotide sequence obtained by sequencing was compared online by BLAST in the NCBI database, and then combined with morphological classification methods to identify the fungal strain V11 as Tyromyceschioneus.

[0042] Example 2

[0043] This embodiment describes a cultivation method for Tyromyces chioneus V11, which includes the following steps:

[0044] S1, Culture medium pretreatment

[0045] The cultivation substrate formula, by weight percentage, is: corn cob 40%, sawdust 43%, wheat bran 15%, gypsum 1%, and lime 1%.

[0046] Weigh the components of the above cultivation formula according to the proportions. Pre-wet the corn cobs and sawdust with clean water 24 hours in advance. After being fully pre-wetted, mix them evenly with auxiliary materials such as bran, gypsum, and lime. The moisture content of the culture medium is 55%.

[0047] S2, Packing and Sterilization

[0048] The cultivation bags are 17cm × 33cm polyethylene plastic bags, filled mechanically and sealed, with sticks inserted manually. Each bag of culture material (wet material) weighs 1.4kg, of which the cultivation substrate (dry material) weighs approximately 0.63kg. The prepared material bags are sterilized at 117℃ for 10 hours, then left to sit in a sterilizer for 2 hours. When the temperature drops below 70℃, the bags are removed from the sterilizer and transferred to a cooling room to cool to below 25℃ before inoculation.

[0049] S3, Inoculation and Culture

[0050] Inoculation was carried out under aseptic conditions, with an inoculation amount of 8% of the weight of the cultivation substrate (dry material). After inoculation, the bags were sealed with sterile cotton plugs. The inoculated bags were placed in the mycelium incubation room and cultured in the dark, with the substrate temperature controlled at 22℃-25℃. After 10 days, the substrate temperature was adjusted to 20℃-22℃. The relative humidity was controlled below 40%. The air was ventilated 1-2 times a day for 0.5 hours each time to keep the air fresh.

[0051] S4, Post-ripening culture

[0052] After the mycelium has fully colonized the bag, it can be cultured for another 5-10 days, with the indoor temperature adjusted to 18℃-22℃.

[0053] S5, Inducing Bud and Mushroom Growth

[0054] Once the mycelium has fully colonized the bags, move them into the fruiting room and arrange them horizontally against the wall on the ground, ensuring close proximity between bags and rows. Stack 4-6 layers, leaving the bag openings and bottoms slightly apart to create two fruiting surfaces. Use a sterilized blade to make a 10mm-20mm long "I"-shaped cut at the bag opening. After opening the bags, allow the mycelium to recover for 2-3 days, lowering the temperature inside the greenhouse to 18℃-22℃. Spray water into the air, greenhouse film, and ground 3-5 times daily, or use a misting system to maintain a relative humidity of 80%-85%. Ensure proper ventilation to keep the air inside the greenhouse fresh. Reduce the amount of covering on the greenhouse roof and increase diffused light appropriately.

[0055] S6, Mushroom Production Management

[0056] After the mushroom buds form, the relative humidity inside the shed should be maintained at 80%-90%, the temperature at 22℃-25℃, the air inside the shed should be kept fresh, and the light intensity should be controlled at 100Lx-400Lx.

[0057] S7 Harvesting

[0058] Harvest once the mushrooms are mature. Stop watering 24 hours before harvesting. Keep your hands and tools clean when picking mushrooms. Gently remove the thin-skinned, cheese-like fruiting bodies from the base, without attaching any substrate.

[0059] S8, Post-harvest Management

[0060] After each harvest, promptly remove any remaining mushrooms and discard any contaminated bags. Generally, withhold water for 3-5 days after harvest to allow the mycelium to resume growth before commencing mushroom cultivation.

[0061] Comparative Example 1

[0062] The cultivation of Tyromyces chioneus V11 followed the same steps as in Example 2, except that the cultivation substrate formula was: 40% soybean straw, 43% sawdust, 15% wheat bran, 1% gypsum, and 1% lime.

[0063] Comparative Example 2

[0064] The cultivation of Tyromyces chioneus V11 followed the same steps as in Example 2, except that the cultivation substrate formula was: 40% corn stalks, 43% sawdust, 15% wheat bran, 1% gypsum, and 1% lime.

[0065] Comparative Example 3

[0066] The cultivation of Tyromyces chioneus V11 followed the same steps as in Example 2, except that the cultivation substrate formula was: 83% sawdust, 15% wheat bran, 1% gypsum, and 1% lime.

[0067] Examples 2 and Comparative Examples 1-3 used corn cobs, soybean straw, corn stalks, sawdust, and wheat bran as the main nutrient sources for the cultivation of Tyromyces chioneus V11. The mycelial growth and development of Tyromyces chioneus V11 under different cultivation substrate formulations are shown in Table 1.

[0068] Table 1. Mycelial growth and development

[0069] deal with Mycelial growth time (d) Time of primordium appearance (d) Tidal transition period (d) Example 2 <![CDATA[30.67±0.58 a ]]> <![CDATA[9.33±0.58 c ]]> <![CDATA[13.00±1.00 a ]]> Comparative Example 1 <![CDATA[26.67±0.58 c ]]> <![CDATA[11.67±0.58 a ]]> <![CDATA[12.67±0.58 ab ]]> Comparative Example 2 <![CDATA[28.00±1.00 b ]]> <![CDATA[10.67±0.58 ab ]]> <![CDATA[11.33±0.58 b ]]> Comparative Example 3 <![CDATA[28.33±1.15 b ]]> <![CDATA[10.33±0.58 bc ]]> <![CDATA[12.33±0.58 ab ]]>

[0070] Note: Results are expressed as mean ± standard deviation. The same lowercase letter indicates no significant difference, while different lowercase letters indicate a significant difference (α = 0.05, ANOVA, LSD method).

[0071] The growth and development of *Cephalotaxus fortunei* mycelium varied under different cultivation substrate formulations (Table 1). The mycelial inoculation time ranged from (26.67±0.58) to (30.67±0.58) days. Example 2 showed the longest inoculation time, significantly delayed compared to Comparative Examples 2 and 3, with no significant difference between the two. Comparative Example 1 showed a significantly faster inoculation time than the other treatments. In this invention, the primordium appearance time (d) is defined as the time from when the mycelium fully colonizes the bag to when the primordia appear. Primordium morphology is shown in [Table 1]. Figure 1 The time of primordia appearance varied among the treatments. Primordia appeared earliest in Example 2, significantly earlier than in the other treatments. Primordia appeared latest in Comparative Example 1 at 11.67±0.58 days, which was not significantly different from Comparative Example 2 (10.67±0.58 days), but significantly longer than Comparative Example 3 and Example 2. The tide transition time was longest in Example 2 and shortest in Comparative Example 2, differing by approximately 2 days. The difference between the two was significant, but neither was significantly different from Comparative Example 1 and Comparative Example 3.

[0072] Table 2 shows the yield and biological efficiency of *Cephalotaxus fortunei* under different culture substrate formulations:

[0073] Table 2 Yield and biological efficiency of *Cephalotaxus fortunei*

[0074]

[0075]

[0076] Note: Results are expressed as mean ± standard deviation. The same lowercase letter indicates no significant difference, while different lowercase letters indicate a significant difference (α = 0.05, ANOVA, LSD method).

[0077] See the morphology of the fruiting body of the thin-skinned cheese fungus. Figure 2 The above results indicate that all treatments successfully produced two flushes of fruiting bodies, with the first flush yielding a higher yield than the second. Overall, Example 2 had the highest total yield, reaching 196.80±5.21 g / bag, with a biological efficiency of 35.14±0.93%, significantly higher than the other three treatments. There were no significant differences among Comparative Examples 1 to 3, with biological efficiencies of 31.14±1.92%, 29.28±0.79%, and 29.63±2.64%, respectively.

[0078] The results showed that, compared to a single substrate, a rationally proportioned composite substrate provided more sufficient nutritional conditions for the growth and development of fungi, thereby increasing yield. Utilizing the biomass degradation ability of *Bacillus thunbergii*, we selected corn cobs, soybean straw, and corn stalks—common agricultural wastes in Northeast China—as cultivation substrates, supplemented with sawdust and wheat bran, and designed a formula according to specific proportions for the cultivation of *Bacillus thunbergii*. This approach not only alleviates environmental pressure but also produces a rich resource of new fungi, achieving two goals at once and turning waste into treasure. The better yield of *Bacillus thunbergii* in the corn cob composite substrate formula is inseparable from the physicochemical properties of corn cobs. Compared to corn stalks, corn cobs have a denser physical structure and stronger water-holding capacity, both of which are key conditions for high yields. Furthermore, corn cobs do not contain the waxy substances found in soybean straw, making them easier for fungi to degrade, thus providing better nutrition for the growth and development of *Bacillus thunbergii*.

[0079] Example 3

[0080] The nutritional components of the thin-skinned casei cultivated in Example 2 were analyzed using the following method:

[0081] (1) Using the fruiting bodies of Tyromyces chioneus V11 produced in Example 2 as the test target, random samples were taken, dried to constant weight, and their nutritional value was measured. The nutritional content of the Tyromyces chioneus samples was tested by Heilongjiang Huace Testing Technology Co., Ltd. The protein content of the fruiting bodies was determined according to GB 5009.5-2016; the dietary fiber content was determined according to GB 5009.88-2023; the calcium content was determined according to GB 5009.92-2016; and the total flavonoid content was determined using the nitrite-aluminum nitrate method.

[0082] (2) Determination of total flavonoid content

[0083] Sample preparation for *Cephalotaxus fortunei* test: Accurately weigh 0.5 g of fruiting body powder and place it in a 50 mL stoppered centrifuge tube. Add 25 mL of methanol, weigh, and sonicate (250 W, 40 kHz) for 30 min. After standing to room temperature, accurately weigh again, make up the weight loss with methanol, shake well, and centrifuge at 10000 r / min for 20 min. Take the supernatant for determining the total flavonoid content. All test indicators were repeated 3 times.

[0084] Determination Method: The total flavonoid content in *Cephalotaxus fortunei* was determined using the nitrite-aluminum nitrate method. 0, 0.2, 0.4, 0.6, 0.8, and 1.0 mL of 0.307 mg / mL rutin standard solution were added to 10.0 mL volumetric flasks. 0.3 mL of 5% NaNO2 was added, mixed, and allowed to stand for 5 min. 0.3 mL of 5% Al(NO3)3 was added, mixed, and allowed to stand for 5 min. 4.0 mL of 4% NaOH was added, and the solution was diluted to 10.0 mL with water, mixed, and allowed to stand for 10 min. The absorbance (A) was measured at 504 nm. A standard curve was constructed by plotting the rutin concentration (mg / mL) on the x-axis and absorbance (A) on the y-axis. The determination was performed in triplicate, and the regression equation was obtained. The absorbance of the *Cephalotaxus fortunei* sample solution was measured at 504 nm, and the total flavonoid concentration (mg / mL) and content in the sample were calculated.

[0085] (3) Test Results

[0086] The nutritional composition of the fruiting body of the thin-skinned cheese mushroom is shown in Table 3. Compared with other edible fungi, the total flavonoid content is relatively high, reaching 708 mg / 100g, which is comparable to that of the medicinal fungus Sanghuang, which is known to have a high flavonoid content (Xie Chunqin, Yang Hetong, Wu Qinyan, et al. Screening experiment of high-yield and excellent strains of Sanghuang flavonoids and polysaccharides. Jiangsu Agricultural Sciences, 2019, 47(14):209-212). The dietary fiber (18.82g / 100g) and calcium content (554mg / kg) are lower than those of black fungus (data not published), and the crude protein content (16.3g / 100g) is lower than that of the flower-faced mushroom (Sheng, CG; Pan, CL; Wang, YF et al. Turn waste intotreasure: Spent substrates of Auricularia heimuer can be used as the substrate for Lepista sordida cultivation. Horticulturae 2023, 9, 1074.).

[0087] Table 3 Nutritional composition analysis of *Cephalotaxus fortunei*

[0088]

[0089] Example 4

[0090] This example demonstrates the degradation of matrix lignocellulose (lignin, cellulose, and hemicellulose) by *Cephalotaxus fortunei*.

[0091] The degradation rate of cellulose, hemicellulose and lignin in the cultivation substrate was investigated for Tyromyces chioneus V11 obtained by the cultivation method in Example 2.

[0092] (1) Measurement method

[0093] The lignin content of the cultivation substrate was determined in accordance with GB / T 20805-2006. The methods for determining and calculating the cellulose and hemicellulose content were based on the instructions of the cellulose content test kit AKSU007C and the hemicellulose content test kit AKSU008C.

[0094] (2) Degradation rate calculation

[0095] Lignin degradation rate (%) = (Lignin content in the original matrix - Lignin content in the fungal residue) / Lignin content in the original matrix * 100%;

[0096] Cellulose degradation rate (%) = (cellulose content in the original substrate - cellulose content in the bacterial residue) / cellulose content in the original substrate * 100%;

[0097] Hemicellulose degradation rate (%) = (hemicellulose content in the original substrate - hemicellulose content in the bacterial residue) / hemicellulose content in the original substrate * 100%.

[0098] (3) Test Results

[0099] Table 4 shows the degradation of matrix lignocellulose by *Cephalotaxus fortunei*. *Cephalotaxus fortunei* has a strong ability to degrade lignocellulose, with a degradation rate of 33.93% for lignin, 37.72% for cellulose, and 28.96% for hemicellulose.

[0100] Table 4. Degradation of matrix lignocellulose by *Cephalotaxus fortunei*

[0101] index Lignin cellulose hemicellulose Original matrix (%) 28.0 45.6 25.9 Fungal residue substrate (%) 18.5 28.4 18.4 Degradation rate (%) 33.93 37.72 28.96

[0102] Contrary to previous understanding that *Cephalotaxus fortunei* possesses a strong ability to degrade lignin, the results of this experiment show that this *Cephalotaxus fortunei* strain has a greater ability to degrade cellulose (37.72%) than lignin (33.93%). Compared to other cultivation substrate materials, corn cobs have lower lignin and higher cellulose content, which corroborates the high yield achieved by *Cephalotaxus fortunei* in corn cob substrate.

[0103] The above description is only a preferred embodiment of the present invention and does not limit the present invention in any way. Any simple modification, change and equivalent variation made to the above embodiment based on the essence of the invention technology shall still fall within the scope of protection of the technical solution of the present invention.

Claims

1. A strain of *Cephalomyces leuciscus*, characterized in that, The strain is Tyromyces chioneus V11, with accession number GDMCC NO.65077.

2. A method for cultivating the thin-skinned casei as described in claim 1, characterized in that, The method includes the following steps: S1, Culture medium pretreatment The cultivation substrate formula by weight percentage is: 40% corn cob, 43% sawdust, 15% wheat bran, 1% gypsum, and 1% lime. Weigh the raw materials according to the proportion, pre-moisten the corn cob and sawdust with clean water 24 hours in advance, and then mix them evenly with wheat bran, gypsum, and lime to obtain the culture medium. S2, Packing and Sterilization The culture medium obtained in S1 is bagged, and the bagged culture medium is sterilized at 117℃ for 10 hours, then left to sit for 2 hours. When the temperature drops below 70℃, the culture medium is removed from the sterilizer and cooled to below 25℃ before inoculation. S3, Inoculation and Culture Inoculate with thin-skinned casein under aseptic conditions, seal the container, and then place it in a colony in the dark for incubation. The substrate temperature is controlled at 22℃-25℃. After 10 days, the substrate temperature is adjusted to 20℃-22℃, and the relative humidity is controlled below 40%. Ventilate 1-2 times a day for 0.5 hours each time. S4, Post-ripening culture After the mycelium of the thin-skinned case fungus has fully colonized the spawn bag, continue to cultivate for 5-10 days, and adjust the indoor temperature to 18℃-22℃. S5, Inducing Bud and Mushroom Growth Move the mushroom bags into the fruiting room and place them closely together on the ground, along the wall. Stack the bags tightly together, in rows, with 4-6 layers. Leave the bag openings and bottoms slightly apart between layers to create two fruiting surfaces. Use a sterilized blade to make a 10-20mm long "I" shaped cut at the bag opening. After opening the bags, allow the mycelium to recover for 2-3 days. Lower the temperature inside the greenhouse to 18-22℃, maintain the relative humidity at 80%-85%, provide ventilation, and increase diffused light. S6, Mushroom Production Management After the mushroom buds form, the relative humidity inside the shed should be maintained at 80%-90%, the temperature at 22℃-25℃, the air inside the shed should be kept fresh, and the light intensity should be controlled at 100Lx-400Lx. S7 Harvesting Harvest once the mushrooms are mature. Stop watering 24 hours before harvesting. Keep your hands and tools clean and hygienic when harvesting. Gently remove the fruiting bodies of the thin-skinned cheese mushroom from the base, without attaching any substrate. S8, Post-harvest Management After each harvest, promptly clean up any remaining mushrooms and remove any contaminated bags. After harvesting, stop watering for 3-5 days to allow the mycelium to resume growth before proceeding with secondary mushroom cultivation management.

3. The cultivation method according to claim 2, characterized in that, The moisture content of the culture medium described in S1 is 55%-60%.

4. The cultivation method according to claim 2, characterized in that, The inoculation amount of the thin-skinned casei described in S3 is 8% of the weight of the cultivation substrate.

5. The use of the thin-skinned casei strain of claim 1 in the degradation of matrix lignin, cellulose and hemicellulose.

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