Soil covering material for industrial cultivation of phlebopus portentosus as well as preparation and application of soil covering material

Abstract

The invention discloses a soil covering material for industrial cultivation of phlebopus portentosus and preparation and application thereof, belongs to the technical field of soil sterilization and edible fungus cultivation, and particularly discloses a preparation method of the soil covering material, the method comprises the following steps: paving mixed soil until the thickness is 3-5cm, exposing for 3-5min under the condition that the microwave frequency is 915MHz-2450MHz, and covering the mixed soil with a soil covering layer; when the material temperature reaches 75-80 DEG C, obtaining a soil covering material; the mixed soil is prepared by uniformly mixing turfy soil and red soil according to a volume ratio of (2: 8)-(8: 2); the water content of the mixed soil is 34%-46%. The obtained soil covering material has the characteristics of good air permeability and low mould content. Furthermore, the earthing material is used for carrying out earthing culture on cultivation fungus bottles or fungus bags of the phlebopus portentosus, and compared with an earthing material obtained by a conventional technology, the yield is obviously improved. The method can be used for agricultural production.

Description

Technical Field

[0001] This invention patent belongs to the field of soil sterilization and edible fungi cultivation technology, specifically relating to a casing material for the industrial cultivation of Boletus edulis and its preparation and application. Background Technology

[0002] Dark brown stalked bolete PHlebopus portentosus (Berk. & Boletus broome (also known as Boletus glomeratus or Boletus galbana), commonly called "black boletus," belongs to the order Boletales, family Boletinellaceae, and genus Phlebopus. It grows wild in tropical and subtropical broad-leaved forests, mainly distributed in Southeast Asia and Yunnan, Guangxi, Guizhou, Sichuan, and Hainan provinces of my country. Currently, Boletus glomeratus is cultivated industrially. Its fruiting bodies are large, fleshy, and delicious, containing high protein, low fat, 18 kinds of amino acids, and abundant essential minerals such as phosphorus, potassium, calcium, magnesium, iron, and zinc, making it highly nutritious. It also contains various bioactive substances such as polysaccharides, polyphenols, sterols, alkaloids, nucleosides, and unsaturated fatty acids, which enhance human immunity, have antioxidant, lipid-lowering, antibacterial, antiviral, antitumor, and anti-ionizing radiation effects. It has long been a popular food and medicinal fungus.

[0003] The industrial cultivation of Boletus edulis requires mulching to produce fruiting bodies. The main functions of mulching are to maintain substrate moisture, regulate gas exchange, stimulate primordia formation, and support fruiting body growth. The mulching material required for industrial cultivation of Boletus edulis must possess properties such as moisture retention, heat preservation, aeration, moderate quality, and resistance to mold contamination. Among these, good aeration and resistance to mold contamination are key factors for achieving fruiting and high yield in industrial cultivation of Boletus edulis. Typically, mushroom casing materials consist of peat moss, humus, garden soil, loam (red, yellow, and brown soils), river mud, and pond mud, which are dried or sun-dried and sieved before use. Sometimes, additives (such as perlite, vermiculite, coconut coir, wheat bran, and rice husks) are added to improve aeration. Since these materials mostly originate from the natural environment, they often carry various harmful molds (such as Trichoderma, Penicillium, Aspergillus, Rhizopus, Mucor, and Neurospora). If used directly as casing material for the industrial cultivation of Boletus thunbergii without treatment, these harmful molds can easily proliferate under the suitable temperature and humidity conditions in the mushroom house. Furthermore, because the mycelium of Boletus thunbergii has very weak resistance to external molds... If the casing material is contaminated with a large amount of mold, it will significantly affect the mycelial growth, mycelial knotting, primordium formation, and fruiting body growth of *Boletus thunbergii* after casing, leading to a substantial reduction in yield. Common disinfection methods for casing materials in edible mushroom cultivation include pasteurization, formaldehyde fumigation, and lime water treatment. However, these methods are unsuitable for treating casing materials used in the industrialized cultivation of *Boletus thunbergii*. Patent CN201510231333 discloses a "Special Antifungal and Yield-Increasing Agent for Casing Material of *Boletus thunbergii*", which involves applying a compound nutrient solution to the casing material of *Boletus thunbergii*. The compound nutrient solution consists of 100-200 g / L potatoes (peeled and boiled), 1-2 g / L magnesium sulfate, and vitamin B1. The mixture consists of 0-20 mg / L of yeast extract, 2-4 g / L of white sugar, 10-20 g / L of potassium dihydrogen phosphate, 1-2 g / L of ferrous sulfate, and 1 L of water. It is sprayed evenly onto the casing material using a spray bottle, ensuring the casing material has a moisture content of 45%–50% and a pH of 5.0–6.5. Patent CN201710136649 discloses "A Casing Formula for *Boletus edulis* and its Preparation Method," characterized by a casing formula consisting of red soil (raw soil) and a nutrient solution. Its technical features lie in first preparing the nutrient solution according to the following steps: 10-20% potatoes (peeled, sliced, and boiled), 1.5-2.0% carbon source, 0.2-0.4% nitrogen source, 0.1-0.2% potassium dihydrogen phosphate, 0.1-0.2% magnesium sulfate, 0.1-0.2% ammonium tartrate, 24 ml / L olive oil, pH 5.5-7.0, with the remainder being water. Then, the prepared nutrient solution is evenly sprayed onto the red soil (raw soil) covering material using a spray bottle, so that its moisture content is 45%-60% and its pH value is 5.5-7.0, the ratio of red soil to nutrient solution should be between 50:1 and 40:1. Both methods aim to enhance the resistance of *Boletus glomeratus* mycelium by increasing the nutrients in the casing material. However, these nutrients can also promote the growth of various harmful molds already present in the casing material. Furthermore, the viscosity of the nutrient solution affects the aeration porosity of the casing material. These combined negative factors ultimately hinder the mycelial knotting, primordia formation, and fruiting body growth of *Boletus glomeratus*, failing to achieve the goal of increasing fruiting yield. Summary of the Invention

[0004] In view of this, and in light of the shortcomings of existing technologies, the purpose of this application is to provide a casing material for the industrial cultivation of *Boletus edulis*, its preparation, and its application. This casing material is composed of peat moss and red soil mixed in an appropriate volume ratio and sterilized using a tunnel-type microwave sterilizer. This invention comprehensively considers the physical and biochemical properties required for the casing material in the industrial cultivation of *Boletus edulis*, taking advantage of the strengths and compensating for the weaknesses: peat moss has good moisture retention and aeration, but when used alone as a casing material, it has the disadvantages of being lightweight and having poor heat retention; red soil has good heat retention and a certain degree of moisture retention. While both are suitable for use as casing materials, their individual weight and poor aeration are drawbacks. A proper volume ratio of both allows for the optimal performance of casing materials in the industrial cultivation of *Boletus edulis*, providing moisture retention, heat preservation, aeration, and moderate weight. Further sterilization of the mixed material using a tunnel-type microwave sterilizer effectively controls mold contamination. This invention also reveals that microwave action causes vibrations in the polar molecules of the mixture, leading to expansion and a looser texture, significantly improving aeration and creating crucial conditions for high-yield, industrialized cultivation of *Boletus edulis*.

[0005] In a first aspect, this application provides a method for preparing a soil covering material, the method comprising spreading a mixed soil to a thickness of 3-5 cm, exposing it to microwave frequencies of 915 MHz-2450 MHz for 3-5 minutes to obtain the soil covering material;

[0006] The mixed soil is prepared by mixing peat soil and red soil in a volume ratio of (2:8)-(8:2);

[0007] The moisture content of the mixed soil is 34%-46%.

[0008] On the other hand, this application provides a soil covering material prepared by any of the methods described above.

[0009] On the other hand, this application provides a cover material, which, by mass percentage, comprises 20%-80% peat soil and 80%-20% red soil, has a porosity of 25.4%-26.7%, and a mold content of 62-91 cfu / g.

[0010] On the other hand, this application provides a method for preparing Boletus thunbergii, the method comprising culturing Boletus thunbergii strains under suitable conditions using a casing material prepared by any of the methods described above or the casing material described above.

[0011] On the other hand, this application provides a dark brown stalked Boletus prepared by the above method.

[0012] Beneficial effects:

[0013] This application discloses a casing material for the industrial cultivation of *Boletus thunbergii*, its preparation, and its application, belonging to the fields of soil sterilization and edible fungi cultivation technology. Specifically, it discloses a method for preparing the casing material, which includes spreading a mixed soil to a thickness of 3-5 cm and exposing it to microwaves at a frequency of 915 MHz-2450 MHz for 3-5 minutes to obtain the casing material. The mixed soil is prepared by mixing peat moss and red soil in a volume ratio of (2:8)-(8:2); the moisture content of the mixed soil is 34%-46%. The obtained casing material has good aeration and low mold content. Further use of this casing material to cultivate *Boletus thunbergii* shows a significantly higher yield compared to casing materials obtained using conventional techniques. It can be used in agricultural production. Attached Figure Description

[0014] Figure 1 This is a graph showing the total number of mold colonies for materials treated differently.

[0015] Figure 2 Porosity diagrams of materials processed at the same time;

[0016] Figure 3 This is a graph showing the mushroom yield. Detailed Implementation

[0017] The specific embodiments of this application will be further described in detail below with reference to the accompanying drawings. These embodiments are only for illustrating this application and are not intended to limit the scope of the invention.

[0018] In the description of this invention, it should be noted that the terms "center," "longitudinal," "lateral," "upper," "lower," "front," "rear," "left," "right," "vertical," "horizontal," "top," "bottom," "inner," and "outer," etc., indicating the orientation or positional relationship, are based on the orientation or positional relationship shown in the accompanying drawings and are only for the convenience of describing this invention and simplifying the description, and do not indicate or imply that the device or element referred to must have a specific orientation, or be constructed and operated in a specific orientation, and therefore should not be construed as a limitation on this invention. Furthermore, the terms "first" and "second" are used for descriptive purposes only and should not be construed as indicating or implying relative importance.

[0019] If the casing material for Boletus thunbergii is sterilized using high-temperature thermal sterilization methods, i.e., using high-temperature, high-pressure steam, it requires a large amount of heat energy and the purchase of large-scale sterilization equipment, resulting in excessively high costs. For a long time, there has been a lack of economical, effective, and scientific options for the casing materials and preparation methods required for the industrial cultivation of Boletus thunbergii.

[0020] The first objective of this application is to provide a method for preparing a casing material with good air permeability, low content of harmful bacteria (especially mold), and high yield of dark brown stalked Boletus.

[0021] In a first aspect, this application provides a method for preparing a cover material, the method comprising spreading a mixed soil to a thickness of 3-5 cm, and then applying it to a microwave environment with a frequency of 915 MHz-2450 MHz and a microwave power of 0.14-0.18 m. 2 In an environment of / kw, obtain cover material;

[0022] The mixed soil is prepared by mixing peat soil and red soil in a volume ratio of (2:8)-(8:2);

[0023] The moisture content of the mixed soil is 34%-46%.

[0024] In a specific embodiment, the exposure time is 3-5 minutes. For example, the exposure time is 3 minutes, 4 minutes, or 5 minutes.

[0025] As used in this application, "peat soil" or "peat soil" refers to organic sediments formed by grasses of the order Poaceae in swampy environments. It consists of incompletely decomposed plant remains, humus, and minerals. It is formed by the accumulation of plant remains under Quaternary anaerobic conditions. Its organic matter content usually exceeds 30%, it is slightly acidic, and has a soft texture. It is mainly distributed in high-latitude cold and humid regions, such as the mountain basins and wetlands of the Changbai Mountains in Northeast China, the Qinghai-Tibet Plateau, and Siberia in Russia. The organic matter content of the peat moss used for the industrial cultivation of Boletus edulis is above 50%, pH 5.0-6.0, and the heavy metal content is: total mercury ≦0.1mg / kg, total arsenic ≦0.8mg / kg, total cadmium ≦0.3mg / kg, and lead ≦1.6mg / kg (refer to Appendix A of NY / T 5010-2016 standard). Red soil is a type of soil that develops under tropical and subtropical rainforests, monsoon forests, or evergreen broad-leaved forests. It is formed by the weathering of carbonate rocks or rocks containing other iron and aluminum oxides under hot and humid climate conditions, and is usually brownish-red. The red soil used for casing in the industrial cultivation of Boletus edulis dark brown stalk comes from most areas south of the Yangtze River and the mountainous areas around the Sichuan Basin. It is loamy and is referred to as red soil in this patent. Its organic matter content is below 5%, pH 5.5-6.5, and it is not contaminated by pesticides or chemicals. Its heavy metal content is as follows: total mercury ≦1.8mg / kg, total arsenic ≦40mg / kg, total cadmium ≦0.3mg / kg, lead ≦90mg / kg, chromium ≦150mg / kg, and copper ≦50mg / kg (refer to GB15618-2018 standard).

[0026] As used in this application, "red soil" refers to zonal soils formed under subtropical and tropical humid climates by intense desilication and aluminization weathering of acidic parent materials such as granite and sandstone / shale. Its core characteristics are a red or brownish-red soil body, rich in iron oxide and aluminum oxide, high clay content, heavy texture, acidic pH (4.5-6.0), low organic matter content, and poor nutrient availability. Classified by parent material, it can be divided into granite red soil, sandstone / shale red soil, and Quaternary red clay red soil. Classified by developmental stage, it can be divided into red soil (typical red soil with moderate desilication and aluminization), lateritic red soil (between red soil and lateritic red soil, in tropical and subtropical transition zones), and yellow-red soil (distributed in higher altitude, higher humidity areas, turning yellow-red due to iron oxide hydration).

[0027] In a specific embodiment, the method for preparing the mixed soil is any one of the following:

[0028] (1) Adjust the moisture content of peat soil and red soil to 34%-46% respectively; mix peat soil and red soil at a volume ratio of (2:8)-(8:2) to obtain the mixed soil;

[0029] (2) Mix peat soil and red soil in a volume ratio of (2:8)-(8:2) to obtain the pre-mixed soil. Adjust the moisture content of the pre-mixed soil to 34%-46% to obtain the mixed soil.

[0030] In a specific embodiment, the method further includes microwave treatment of the mixed soil using a tunneling microwave method.

[0031] In a specific embodiment, the power of the microwave is 0.14-0.18m. 2 / kw.

[0032] In a specific embodiment, the method further includes an outlet temperature of 75-80℃.

[0033] In a specific embodiment, the volume ratio is (2:8)-(3.5:6.5), (2:8)-(4:6), (3.5:6.5)-(4:6), (3.5:6.5)-(2:8), or (4:6)-(8:2).

[0034] In specific embodiments, the moisture content is 34%-38%, 38%-42%, 40%-44%, or 42%-46%. In specific embodiments, the moisture content can be 34%, 35%, 36%, 37%, 38%, 39%, 40%, 41%, 42%, 43%, 44%, 45%, or 46%.

[0035] In a specific embodiment, the mixing is accompanied by stirring, and preferably, the stirring time is 5-10 minutes.

[0036] In a specific embodiment, the porosity of the cover material is 25.4%-26.7%; in a specific embodiment, the porosity is 26.1%.

[0037] In a specific embodiment, the porosity of the covering material can be 25.4%, 25.5%, 25.6%, 25.7%, 25.8%, 25.9%, 26.0%, 26.1%, 26.2%, 26.3%, 26.4%, 26.5%, 26.6%, or 26.7%.

[0038] In a specific embodiment, the mold content of the covering material is 62-91 cfu / g.

[0039] In specific embodiments, the mold content can be 62cfu / g, 63cfu / g, 64cfu / g, 65cfu / g, 66cfu / g, 67cfu / g, 68cfu / g, 69cfu / g, 70cfu / g, 71cfu / g, 72cfu / g, 73cfu / g, 74cfu / g, 75cfu / g, 76cfu / g, 77cfu / g, 78cfu / g, 79cfu / g, 80cfu / g, 81cfu / g, 82cfu / g, 83cfu / g, 84cfu / g, 85cfu / g, 86cfu / g, 87cfu / g, 88cfu / g, 89cfu / g, 90cfu / g, 91cfu / g.

[0040] In specific embodiments, the volume ratio of peat soil to red soil is 4:6, 2:8, 4:6, 8:2, or 3.5:6.5.

[0041] In a specific embodiment, the moisture content is 38%-42%, 34%-38%, 38%-42%, 42%-46%, or 40%-44%.

[0042] In a specific embodiment, the stirring time is 5-10 minutes.

[0043] In a specific embodiment, the stirring time can be 5 minutes, 6 minutes, 7 minutes, 8 minutes, 9 minutes, or 10 minutes.

[0044] In specific embodiments, the thickness is 3cm, the thickness is 4cm, and the thickness is 5cm.

[0045] In a specific embodiment, the microwave frequency is 2450MHz or the microwave frequency is 915MHz.

[0046] In a specific embodiment, the microwave time is 5 minutes, 4 minutes, or 3 minutes.

[0047] In a specific embodiment, the discharge temperature is 75-80℃.

[0048] For example, the volume ratio of peat soil to red soil is 4:6; the moisture content is 40%; the stirring time is 5-10 minutes; the thickness is 4 cm; the microwave frequency is 2450 MHz; and the microwave power output distribution is 0.16 m. 2 / kw; the microwave time is 4min; the discharge temperature is 78℃.

[0049] For example, the volume ratio of peat soil to red soil is 2:8; the moisture content is 46%; the stirring time is 5-10 minutes; the thickness is 3 cm; the microwave frequency is 2450 MHz; and the microwave power output distribution is 0.18 m. 2 / kw; the microwave time is 5min; the discharge temperature is 80℃.

[0050] For example, the volume ratio of peat soil to red soil is 4:6; the moisture content is 38%; the stirring time is 5-10 minutes; the thickness is 4 cm; the microwave frequency is 2450 MHz; the microwave time is 4 minutes; and the microwave power output distribution is 0.15 m. 2 / kw; the discharge temperature is 77℃.

[0051] For example, the volume ratio of peat soil to red soil is 8:2; the moisture content is 34%; the stirring time is 5-10 minutes; the thickness is 5 cm; the microwave frequency is 2450 MHz; the microwave time is 3 minutes; and the microwave power output distribution is 0.14 m. 2 / kw; the discharge temperature is 75℃.

[0052] For example, the volume ratio of peat soil to red soil is 3.5:6.5; the moisture content is 42%; the stirring time is 5-10 minutes; the thickness is 5 cm; the microwave frequency is 915 MHz; the microwave time is 4 minutes; and the microwave power output distribution is 0.17 m. 2 / kw; the discharge temperature is 76℃.

[0053] On the other hand, this application provides a soil covering material prepared by the method described above.

[0054] On the other hand, this application provides a cover material, which, by mass percentage, comprises 20%-80% peat soil and 80%-20% red soil, has a porosity of 25.4%-26.7%, and a mold content of 62-91 cfu / g.

[0055] This invention provides a casing material for the industrial cultivation of Boletus edulis, its preparation, and its application. This casing material is a mixture of peat moss and red soil in an appropriate volume ratio, and is sterilized using a tunnel-type microwave sterilizer. This invention comprehensively considers the physical and biochemical properties required for the industrial cultivation of *Boletus edulis*, combining their strengths and compensating for their weaknesses: peat moss has good moisture retention and aeration, but as a casing material alone, it is lightweight and has poor heat retention; red soil has good heat retention and some moisture retention, but as a casing material alone, it is heavy and has poor aeration. The appropriate volume ratio of the two materials satisfies the requirements for moisture retention, heat retention, aeration, and moderate weight for the industrial cultivation of *Boletus edulis*. The mixture is then sterilized using a tunnel-type microwave sterilizer, effectively controlling mold contamination. This invention also reveals that the microwave action causes the polar molecules in the mixture to vibrate, resulting in a swelling effect that loosens the mixture and significantly improves its aeration, creating important conditions for the industrial cultivation and high yield of *Boletus edulis*.

[0056] Microwave sterilization technology principle: Microwaves are high-frequency electromagnetic waves with a frequency of 300MHz to 300GHz. The sterilization effect of microwaves is the result of the combined action of microwave thermal and non-thermal effects. Microwaves can force polar molecules (especially water molecules) in materials to vibrate and rotate at extremely high frequencies, thereby generating a large amount of heat inside the material. This damages the cell structure of mold in the material, causing protein denaturation and death. Microwaves can also interfere with the spatial structure and function of biological macromolecules such as DNA, RNA, and proteins in mold cells, affecting cell membrane permeability and electrical potential, leading to abnormal cell metabolism, inhibited growth and development, and death. Microwave sterilization is short in time, fast in speed, and provides uniform heating, enabling rapid heating and sterilization of the interior of the casing material. Microwave sterilization is a low-temperature sterilization method. Compared with conventional thermal sterilization, it can achieve the desired sterilization effect at a lower temperature and in a shorter time. Generally, the sterilization temperature is 75℃-80℃, and the sterilization time is 3-5 minutes to achieve the effect of killing mold, while conventional thermal sterilization methods require a temperature of 121℃-132℃ and a time of about 1-2 hours. The tunnel-type microwave sterilizer can work continuously, has a large material processing capacity, and high working efficiency, which can meet the needs of the casing material for the industrial cultivation of Boletus thunbergii. Microwave sterilization technology is a physical technology that is applied to the treatment of casing material for the industrial cultivation of Boletus thunbergii, without the risk of other chemical residues.

[0057] The covering material of this invention is composed of a mixture of peat moss and red soil, wherein the peat moss accounts for 20%-80% by volume and the red soil accounts for 80%-20% by volume. First, the peat moss and red soil are sieved separately (1cm×1cm sieve holes) to remove larger fragments, particles, and impurities. Then, they are sprayed with water to moisten them, controlling the moisture content to be 35%-50%. Next, the two materials are added to the same mixer at a suitable volume ratio and stirred for 5-10 minutes until uniformly mixed. The mixture is then discharged and fed into the feed hopper of a tunnel-type microwave sterilizer. In the process, spread the mixture evenly on the material conveyor belt of the tunnel microwave sterilizer, adjust the thickness of the mixture to 3-5cm, select the microwave frequency of 2450MHz or 915MHz, adjust the conveyor speed, and control the heating time of the mixture in the microwave sterilizer to 3-5 minutes. The mixture is output from the discharge end of the tunnel microwave sterilizer at a temperature of 75-80℃. After the mixture exits the machine, immediately transfer it to a clean iron drum, cover it tightly with a clean plastic sheet, and let it cool to room temperature before using it as a casing material for the industrial cultivation of Boletus edulis.

[0058] Compared with existing microwave sterilization casing materials, the mushroom yield of Boletus thunbergii cultured using the casing material prepared in this application is significantly increased, with a yield increase rate of 35.6%-38.9% on day 15, and a maximum yield of 531.7 kg / 5000 kg (casing material).

[0059] Furthermore, this application provides a method for cultivating Boletus thunbergii using the aforementioned casing material.

[0060] On the other hand, this application provides a method for preparing Boletus thunbergii, the method comprising culturing Boletus thunbergii strains under suitable conditions using a casing material prepared by any of the methods described above or the casing material described above.

[0061] In a specific embodiment, the suitable conditions include a temperature of 25.5℃-29.5℃, a relative humidity of 90%-99%, a carbon dioxide concentration of 0.1%-0.3%, and a diffuse white light of 100-300 lux.

[0062] In a specific embodiment, the temperature may be 25.5℃, 26.0℃, 26.5℃, 27.0℃, 28.0℃, 28.5℃, 29.0℃, or 29.5℃.

[0063] In specific embodiments, the relative humidity may be 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%.

[0064] In specific embodiments, the carbon dioxide concentration may be 0.1%, 0.2%, or 0.3%.

[0065] In a specific embodiment, the scattered white light can be 100 lux, 150 lux, 200 lux, 250 lux, or 300 lux.

[0066] In a specific embodiment, the Boletus thunbergii strain is selected from at least one of its spores, mycelium and fruiting body;

[0067] In a specific embodiment, the culture time is at least 15 days. In a specific embodiment, the culture time is 15 days, 16 days, 17 days, 18 days, 19 days, 20 days, ... 100 days.

[0068] In a specific embodiment, the method includes covering the culture bottles or bags of *Boletus edulis* with soil, wherein the soil covering thickness is 2-3 cm.

[0069] In a specific embodiment, the soil covering thickness is 2cm or 3cm.

[0070] On the other hand, this application provides a dark brown stalked Boletus prepared by the above method.

[0071] In a specific embodiment, the mass ratio of the yield of *Boletus edulis* to the cultivation substrate material is >382.9:5000. In a specific embodiment, the mass ratio of the yield to the cultivation substrate material is ≥519.3:5000; in a specific embodiment, the mass ratio of the yield to the cultivation substrate material is ≥520.8:5000; in a specific embodiment, the mass ratio of the yield to the cultivation substrate material is ≥524.2:5000; in a specific embodiment, the mass ratio of the yield to the cultivation substrate material is ≥531.7:5000. For example, the yield is based on a total yield of 4000 bottles. Further example, each bottle has a volume of 1.5L. Further example, the bottle mouth diameter is 9cm. Further example, the bottle mouth height is 4.5cm.

[0072] In a specific embodiment, the yield refers to the fruiting body yield. In a specific embodiment, the fruiting body yield refers to the above-ground portion yield.

[0073] On the other hand, this application provides the use of casing materials prepared by any of the methods described above, or the use of the aforementioned casing materials in the cultivation of Boletus thunbergii.

[0074] Therefore, it can be seen that the *Boletus edulis* var. *dark brown stalk* cultivated using the above-mentioned casing material has a high fruiting yield, with the yield-to-mass ratio of the cultivation substrate material reaching as high as 531.7:5000. It can be used in agricultural production.

[0075] For example, before covering with soil, a sample of this mixture was taken using a sterile tube and sent to a microbiology testing laboratory. The sample was incubated on PDA medium at 28°C for 5 days using the dilution plate count method. The total number of mold colonies was then detected and counted. The specific procedure is as follows:

[0076] Sample preparation and dilution: Weigh 10 grams of the mixed sample, add 90 ml of sterile water and shake thoroughly to prepare a 1:10 homogenate; then, use a sterile pipette to take 1 ml of the above homogenate and add it to 9 ml of sterile water to prepare a 1:100 dilution. Repeat this process to prepare a series of gradient dilutions (e.g., 10:100). -3 10 -4 wait);

[0077] Inoculation and culture: Select the above-mentioned appropriate concentration of dilution, take 0.1 ml to 1 ml of each dilution and inoculate it onto a plate containing PDA medium. Invert the plate in a constant temperature incubator at 28℃ and culture for 5-7 days.

[0078] Counting and Calculation: Select plates with colony counts between 30-300 CFU for counting. Calculate the total number of mold colonies in the sample using the following formula.

[0079] Total number of mold colonies (CFU / g) = average number of colonies on plate × dilution factor / inoculum volume (ml) × sample mass (g).

[0080] Before backfilling, the aeration porosity of the backfill material was determined using a ring sampler, in accordance with the national standard NY / T2118-2012. The specific operation process is as follows:

[0081] Prepare a ring cutter (volume 100cm³) 3 );

[0082] Secure the bottom of the ring cutter with the bottom cap without holes, fill the top with the air-dried soil sample from the top, and then close the top cap with holes. The compaction of the sample should be close to the compaction state when the soil was being used for backfilling.

[0083] With the perforated top cap facing upwards, immerse the ring cutter in a water-filled plastic square box for 24 hours. After removing it, wipe the water off the outer surface of the ring cutter with absorbent paper and weigh it immediately (W1). During immersion, the water level should be 2cm above the top cap of the ring cutter.

[0084] Place the perforated top of the ring sampler face down on a funnel lined with filter paper. Let it stand for 3 hours. Collect the water that has freely drained from the sample using a clean beaker until no more water seeps out. Weigh the ring sampler, the soil sample, and the total weight of the water it holds (W2). Calculate as follows:

[0085] Ventilation porosity (%) = W1–W2 / 100.

[0086] Example 1

[0087] Sieve peat moss and red soil separately (1cm x 1cm sieve holes) to remove larger fragments, particles, and impurities. Measure out portions of peat moss and red soil, with peat moss comprising 40% and red soil 60% by volume. Spray each portion with water to moisten it, controlling the moisture content to 40%. Then, add both materials to the same mixer and mix for 5-10 minutes until homogeneous. Discharge the mixture to obtain a final product. Use a sterile tube to collect the mixture and send it to a microbiology laboratory. Incubate the mixture on PDA medium at 28°C for 5 days using the dilution plate count method to perform a total mold colony count. The specific procedure is as follows:

[0088] Sample preparation and dilution: Weigh 10 grams of the mixed sample, add 90 ml of sterile water and shake thoroughly to prepare a 1:10 homogenate; then, use a sterile pipette to take 1 ml of the above homogenate and add it to 9 ml of sterile water to prepare a 1:100 dilution. Repeat this process to prepare a series of gradient dilutions (e.g., 10:100). -3 10 -4 wait);

[0089] Inoculation and culture: Select the above-mentioned appropriate concentration of dilution, take 0.1 ml to 1 ml of each dilution and inoculate it onto a plate containing PDA medium. Invert the plate and place it in a constant temperature incubator at 28℃ for 5 to 7 days.

[0090] Counting and Calculation: Select plates with colony counts between 30 and 300 CFU for counting, and calculate the total number of mold colonies in the sample using the following formula;

[0091] Total number of mold colonies (CFU / g) = average number of colonies on plate × dilution factor / inoculum volume (ml) × sample mass (g).

[0092] The results are shown in Table 1 for the mixed treatment of peat soil (40%), red soil (60%), and... Figure 1 The mixing process is shown in the figure.

[0093] Simultaneously, take the mixed materials, air dry them, and then use a volume of 100cm³. 3 The ring sample was used to determine the ventilation porosity (refer to national standard NY / T2118-2012). The specific operation procedure is as follows:

[0094] Prepare a ring cutter (volume 100cm³) 3 );

[0095] Secure the bottom of the ring cutter with the bottom cap without holes, fill the top with the air-dried soil sample from the top, and then close the top cap with holes. The compaction of the sample should be close to the compaction state when the soil was being used for backfilling.

[0096] With the perforated top cap facing upwards, immerse the ring cutter in a water-filled plastic square box for 24 hours. After removing it, wipe the water off the outer surface of the ring cutter with absorbent paper and weigh it immediately (W1). During immersion, the water level should be 2cm above the top cap of the ring cutter.

[0097] Place the perforated top of the ring cutter face down on a funnel lined with filter paper. Let it stand for 3 hours. Collect the water that has freely drained from the sample using a clean beaker until no more water seeps out. Weigh the total weight (W2) of the ring cutter, the mixture, and the water it holds. Calculate as follows:

[0098] Ventilation porosity (%) = W1–W2 / 100.

[0099] The results are shown in Table 2 for the mixed treatment of peat soil (40%), red soil (60%), and... Figure 2 The mixing process is shown in the figure.

[0100] The mixture is then fed into the feed hopper of the tunnel-type microwave sterilizer, ensuring it is evenly spread onto the conveyor belt. The thickness of the mixture is adjusted to 4cm, the microwave frequency is selected as 2450MHz, and the microwave power output distribution is set to 0.16m. 2 / kw, adjust the conveying speed so that the heating time of the mixture in the microwave sterilizer is 4 minutes. The mixture is output from the discharge end of the tunnel microwave sterilizer at a discharge temperature of 78℃. After the mixture is discharged from the machine, it is immediately transferred into a clean iron drum, covered with a clean plastic sheet, and cooled to room temperature to obtain the mixed + microwave treated material.

[0101] The mixed and microwave-treated materials were collected using sterile tubes and sent to the microbiology laboratory. The dilution plate count method was used to incubate the mixture on PDA medium at 28°C for 5 days. A total mold colony count was then performed. The results are shown in Table 1. The mixture of peat moss (40%) and red clay (60%) was microwave-treated. Figure 1 The mixture and microwave treatment is shown in the figure. Simultaneously, the mixed and microwave-treated material is air-dried and then used in a 100cm³ volume container. 3 The aeration porosity was determined using a ring sampler, and the results are shown in Table 2. The mixture of peat soil (40%) and laterite (60%) was subjected to microwave treatment. Figure 2 The mixture and microwave processing is shown in the figure. The above experiment was repeated 3 times, and the data statistics are shown in Table 1 and Table 2. Figure 1 The results showed that the total number of mold colonies in the mixture of peat moss (40%) and red soil (60%) after microwave treatment was significantly different from that of the mixed treatment. Figure 2 The results showed that the air porosity of the mixture after microwave treatment of peat moss (40%) and red soil (60%) was significantly different from that of the mixed treatment.

[0102] Table 1 Comparison of total mold colony counts

[0103]

[0104] Table 2 Comparison of ventilation porosity

[0105]

[0106] Example 2

[0107] Sieve the peat moss and red soil separately (1cm x 1cm sieve holes) to remove larger fragments, particles, and impurities. Measure out portions of peat moss and red soil, with peat moss comprising 20% ​​and red soil 80% by volume. Moisten each portion with water, controlling the moisture content to 46%. Then, add both materials to the same mixer and mix for 5-10 minutes until homogeneous. Discharge the mixture and feed it into the feed hopper of a tunnel-type microwave sterilizer, spreading it evenly on the conveyor belt. Adjust the spread thickness to 3cm, select a microwave frequency of 2450MHz, and a microwave power output distribution of 0.18m. 2 / kw, adjust the conveying speed to heat the mixture in the microwave sterilizer for 5 minutes. The mixture is output from the discharge end of the tunnel microwave sterilizer at a discharge temperature of 80°C. After the mixture is discharged, it is immediately transferred to a clean iron drum, covered with a clean plastic sheet, and cooled to room temperature to obtain the casing material for the industrial cultivation of dark brown stalked Boletus in Example 2 (also known as the casing material of Example 2).

[0108] Example 3

[0109] Sieve the peat moss and red soil separately (1cm x 1cm sieve holes) to remove larger fragments, particles, and impurities. Measure out portions of peat moss and red soil, with peat moss comprising 40% and red soil 60% by volume. Spray each portion with water to moisten it, controlling the moisture content to 38%. Then, add both materials to the same mixer and mix for 5-10 minutes until homogeneous. Discharge the mixture and feed it into the feed hopper of a tunnel-type microwave sterilizer, spreading it evenly on the conveyor belt. Adjust the spread thickness to 4cm, select a microwave frequency of 2450MHz, and a microwave power output distribution of 0.15m. 2 / kw, adjust the conveying speed so that the heating time of the mixture in the microwave sterilizer is 4 minutes. The mixture is output from the discharge end of the tunnel microwave sterilizer with an discharge temperature of 77°C. After the mixture is discharged from the machine, it is immediately transferred into a clean iron drum, covered with a clean plastic sheet, and cooled to room temperature to obtain the casing material for the industrial cultivation of dark brown stalked Boletus in Example 3 (also known as the casing material of Example 3).

[0110] Example 4

[0111] Sieve the peat moss and red soil separately (1cm x 1cm sieve holes) to remove larger fragments, particles, and impurities. Measure out portions of peat moss and red soil, with peat moss comprising 80% and red soil 20% by volume. Moisten each portion with water, controlling the moisture content to 34%. Then, add both materials to the same mixer and mix for 5-10 minutes until homogeneous. Discharge the mixture and feed it into the feed hopper of a tunnel-type microwave sterilizer, spreading it evenly on the conveyor belt. Adjust the spread thickness to 5cm, select a microwave frequency of 2450MHz, and a microwave power output distribution of 0.14m. 2 / kw, adjust the conveying speed so that the heating time of the mixture in the microwave sterilizer is 3 minutes. The mixture is output from the discharge end of the tunnel microwave sterilizer with an discharge temperature of 75°C. After the mixture is discharged from the machine, it is immediately transferred into a clean iron drum, covered with a clean plastic sheet, and cooled to room temperature to obtain the casing material for the industrial cultivation of dark brown stalked Boletus in Example 4 (also known as the casing material of Example 4).

[0112] Example 5

[0113] Sieve the peat moss and red soil separately (1cm x 1cm sieve holes) to remove larger fragments, particles, and impurities. Measure out portions of peat moss and red soil, with peat moss comprising 35% and red soil 65% by volume. Moisten each portion with water, controlling the moisture content to 42%. Then, add both materials to the same mixer and mix for 5-10 minutes until homogeneous. Discharge the mixture and feed it into the feed hopper of a tunnel-type microwave sterilizer, spreading it evenly on the conveyor belt. Adjust the spread thickness to 5cm and the microwave power output distribution to 0.17m. 2 / kw, select the microwave frequency as 915MHz, adjust the conveying speed so that the heating time of the mixture in the microwave sterilizer is 4 minutes, the mixture is output from the discharge end of the tunnel microwave sterilizer, the discharge temperature is 76℃, the mixture is immediately transferred into a clean iron drum after exiting the machine, covered tightly with a clean plastic cloth, and cooled to room temperature to obtain the casing material for the industrial cultivation of dark brown stalked Boletus in Example 5 (also known as the casing material of Example 5).

[0114] Comparative Example 1

[0115] Sieve the peat moss and red soil separately (1cm×1cm sieve holes) to remove larger fragments, particles and impurities. Measure out the peat moss and red soil, with peat moss accounting for 40% of the volume and red soil accounting for 60% of the volume. Spray them with water to moisten them, controlling the moisture content of the materials to be between 38% and 42%. Then put the two materials into the same mixer and mix for 5-10 minutes to make them evenly mixed. Discharge the mixture to obtain the comparative proportion of casing material for the industrial cultivation of Boletus edulis (also known as comparative proportion casing material).

[0116] The Boletus edulis culture bottles (polypropylene plastic bottles, 1.5L volume, 9cm mouth diameter, 4.5cm height) that have been inoculated with Boletus edulis spores and cultured at 28-32℃ for 25-30 days, with mycelium fully grown and free from other mold contamination, are taken from the factory's culture room and transported through a clean channel to the casing room. After opening the bottle mouths, the casing materials of Examples 2, 3, 4 and Comparative Example 1 are used on the casing machine to cover the bottles, controlling the casing layer thickness to be 2-3cm. Furthermore, the casing material described in Example 5 was used to casing the cultivation bags of *Boletus pumilus*. The cultivation bags (polypropylene plastic bags, 1.5L volume, 9cm diameter opening, 4.5cm height) containing *Boletus pumilus* spawn and cultured at 28-32℃ for 25-30 days, with mycelial growth covering the entire bag and free from other mold contamination, were taken from the factory's incubation room and transported through a clean channel to the casing room. After opening the bags, they were casing on a casing machine, with the casing layer thickness controlled at 2-3cm. The average weight of each bottle or bag of substrate was 1.25kg, and the total weight of 4000 bottles (bags) was 5000kg.

[0117] In Examples 2, 3, 4, and Comparative Example 1, 4000 bottles of soil were used for covering, and in Example 5, 4000 bags of soil were used. After covering, all were transferred to the same fruiting room and managed under the same environmental conditions. The fruiting conditions in the fruiting room were controlled as follows: temperature 25.5℃-29.5℃, relative humidity 90%-99%, carbon dioxide concentration 0.1%-0.3%, and diffused white light 100-300 lux. After 15 days, the dark brown stamen-like Boletus fruiting bodies matured, and the mushrooms were harvested. The fruiting yield of each example and comparative example was recorded. The three fruiting rooms were repeated. The fruiting yield refers to the fruiting body yield. In this application, the fruiting body yield refers to the portion of the mushroom body growing on the soil covering layer.

[0118] The results are shown in Table 3 and Figure 3 ( Figure 3 In the figures, Comparative Examples 1, 2, 3, 4 and 5 are respectively the soil covering materials for the comparative examples, the soil covering materials for Example 2, the soil covering materials for Example 3, the soil covering materials for Example 4 and the soil covering materials for Example 5. Figure 3The results show that when peat moss and red soil are mixed in an appropriate volume ratio and sterilized using a tunnel microwave sterilizer, the resulting mushroom yield is significantly different from that of the control group when used as the casing material for the industrial cultivation of Boletus edulis.

[0119] Table 3 Comparison of fruiting yields of different embodiments

[0120]

[0121] The above description is only a preferred embodiment of the present invention. It should be noted that for those skilled in the art, several improvements and substitutions can be made without departing from the technical principles of the present invention, and these improvements and substitutions should also be considered within the scope of protection of the present invention.

Claims

1. A method for preparing a soil covering material, characterized in that, The method includes: spreading the mixed soil to a thickness of 3-5 cm, and exposing it to microwaves with a frequency of 915 MHz-2450 MHz and a power of 0.14-0.18 m. 2 Under an environment of / kw, obtain the covering material; The mixed soil includes peat soil and red soil, and the volume ratio of peat soil to red soil is (2:8)-(8:2). The moisture content of the mixed soil is 34%-46%.

2. The method as described in claim 1, characterized in that, The method for preparing the mixed soil is any one of the following: (1) Adjust the moisture content of the peat soil and the red soil to 34%-46% respectively; mix the peat soil and the red soil at a volume ratio of (2:8)-(8:2) to obtain the mixed soil; (2) Mix the peat soil and the red soil at a volume ratio of (2:8)-(8:2) to obtain the pre-mixed soil, and adjust the moisture content of the pre-mixed soil to 34%-46% to obtain the mixed soil.

3. The method as described in claim 1 or 2, characterized in that, The method also includes a discharge temperature of 75-80℃; And / or, the exposure time is 3-5 minutes; And / or, the volume ratio of the peat soil and the red soil is (2:8)-(3.5:6.5), (2:8)-(4:6), (3.5:6.5)-(4:6), (3.5:6.5)-(2:8) or (4:6), (8:2); And / or, the mixing is accompanied by stirring, preferably, the stirring time is 5-10 min.

4. The method according to any one of claims 1-3, characterized in that, The porosity of the cover material is 25.4%-26.7%; And / or, the mold content of the covering material is 62-91 cfu / g.

5. A soil covering material prepared by the method described in any one of claims 1-4.

6. A soil covering material, characterized in that, The covering material includes peat soil and red soil. By mass percentage, the peat soil accounts for 20%-80%, the red soil accounts for 80%-20%, and the porosity of the covering material is 25.4%-26.7%.

7. A method for preparing a dark brown stalked Boletus, characterized in that, The method includes: culturing Boletus glomeratus strain under suitable conditions using a casing material prepared by any one of the methods of claims 1-4, or a casing material of claim 5, or a casing material of claim 6.

8. The method as described in claim 7, characterized in that, The suitable conditions include a temperature of 25.5℃-29.5℃, a relative humidity of 90%-99%, a carbon dioxide concentration of 0.1%-0.3%, and a diffuse white light concentration of 100-300 lux. And / or, the *Boletus glomeratus* species is selected from at least one of its spores, mycelium and fruiting body; And / or, the culture time is at least 15 days; And / or, the method includes covering the culture bottles or bags of *Boletus edulis* with soil, the covering thickness being 2-3 cm.

9. The method as described in claim 7 or 8, characterized in that, The mass ratio of the yield of *Boletus edulis* to the cultivation substrate material is >382.9:5000. Preferably, the mass ratio is ≥519.3:5000. More preferably, the mass ratio is ≥520.8:5000. More preferably, the mass ratio is ≥524.2:5000. More preferably, the mass ratio is ≥531.7:5000.

10. The use of the casing material prepared by any of the methods described in claims 1-4 or the casing material described in claim 5 or 6 in the cultivation of Boletus glomeratus.

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