Cultivation Substrate for Leccinum roseonervosum and Its Preparation Method

Abstract

The present invention discloses a cultivation substrate for Boletus rhodopunctus and a preparation method thereof, belonging to the technical field of bolete cultivation. The cultivation substrate for Boletus rhodopunctus, the composition components of which are by mass percentage: at least one of fresh oats, fresh corn kernels and fresh wheat grains 3-8%, fresh grains 20-35%, soil 20-30%, fresh sawdust and / or corncobs 15-25%, silkworm excrement 1-5%, rice hulls 8-15%, FeSO4 0.005-0.015%, MgSO4 0.05-0.12%, MnSO4 0.01-0.03%, peat soil 3-7%, limestone powder 0.5-2%, EM bacterial agent 0.02-0.1%, high-temperature resistant α-amylase 0.02-0.08%. Using the cultivation substrate of the present invention can effectively solve the problems of difficult artificial cultivation of Boletus rhodopunctus, high contamination rate of fungus bags and low fruiting rate.

Description

Technical Field

[0001] The present invention belongs to the technical field of cultivation technology of bolete fungi, and particularly relates to a cultivation substrate for Pulveroboletus rhodoxanthus and a preparation method thereof. Background Art

[0002] Bolete mushrooms are popular among consumers around the world due to their rich nutrition and delicious taste. Since most bolete mushrooms are symbiotic fungi and need to symbiose with host plants to complete their life cycle, it is difficult to cultivate them artificially. At present, except for Phlebopus portentosus and Sinoboletus ganbajun, there are no reports of successful artificial cultivation of other bolete mushrooms. Phlebopus portentosus is a bolete mushroom variety that can fruit under mushroom house conditions using saprophytic cultivation methods and can be produced on a large scale throughout the year. Pulveroboletus rhodoxanthus contains rich protein, crude fat, total sugar, crude fiber, various mineral elements and amino acids, and is an edible mushroom with relatively high nutritional value. Pulveroboletus rhodoxanthus is deeply loved by consumers because of its delicious taste and bright color. Pulveroboletus rhodoxanthus has a narrow distribution range, is mixed with Phlebopus portentosus in the same habitat of loquat orchards, has a scarce yield, belongs to rare and endangered species, and there are no reports on other distribution areas and research on this strain.

[0003] Pulveroboletus rhodoxanthus is a newly discovered new variety with cultivation potential. The mycelium of Pulveroboletus rhodoxanthus grows slowly, has a lot of secretions, the culture medium is severely browned, germinates slowly on the cultivation substrate, the mycelium grows slowly, the growth cycle is long, the pollution rate is high, the fruiting rate is low, the growth conditions are harsh, and the cultivation is difficult. The large-scale cultivation of Pulveroboletus rhodoxanthus can not only effectively preserve the germplasm resources of the characteristic Pulveroboletus rhodoxanthus, but also bring great economic and social benefits. Summary of the Invention

[0004] The technical problem to be solved by the present invention is the problem that the artificial cultivation of Pulveroboletus rhodoxanthus is difficult, the growth cycle of the strain on the cultivation substrate is long, the pollution rate of the mushroom bags is high, and the fruiting rate is not high.

[0005] The technical solution adopted by the present invention to solve its technical problem is: the cultivation substrate for Pulveroboletus rhodoxanthus, the composition components of which are in mass percentage: at least one of fresh oats, fresh corn kernels and fresh wheat grains is 3-8%, fresh grains are 20-35%, soil is 20-30%, fresh wood chips and / or corncobs are 15-25%, silkworm excrement is 1-5%, rice hulls are 8-15%, FeSO4 is 0.005-0.015%, MgSO4 is 0.05-0.12%, MnSO4 is 0.01-0.03%, peat soil is 3-7%, limestone powder is 0.5-2%, EM microbial agent is 0.02-0.1%, and heat-resistant α-amylase is 0.02-0.08%.

[0006] Furthermore, at least one of fresh oats, fresh corn kernels and fresh wheat grains accounts for 4-6%, fresh grains account for 25-32%, soil accounts for 23-28%, fresh sawdust and / or corncobs account for 18-22%, silkworm excrement accounts for 3-5%, chaff accounts for 10-12%, FeSO4 accounts for 0.008-0.012%, MgSO4 accounts for 0.08-0.12%, MnSO4 accounts for 0.01-0.02%, peat soil accounts for 4-6%, limestone powder accounts for 0.8-1.5%, EM bacterial agent accounts for 0.02-0.05%, and heat-resistant α-amylase accounts for 0.04-0.06%.

[0007] Even further, at least one of fresh oats, fresh corn kernels and fresh wheat grains accounts for 5%, fresh grains account for 30%, soil accounts for 25%, fresh sawdust and / or corncobs account for 20%, silkworm excrement accounts for 3.8%, chaff accounts for 10%, FeSO4 accounts for 0.01%, MgSO4 accounts for 0.1%, MnSO4 accounts for 0.01%, peat soil accounts for 5%, limestone powder accounts for 1%, EM bacterial agent accounts for 0.03%, and heat-resistant α-amylase accounts for 0.05%.

[0008] The above-mentioned fresh grains are fresh millet.

[0009] The particle size of the above-mentioned soil is 0.1-1 cm, the particle size of fresh sawdust and / or corncobs is 0.5-2 cm, the particle size of peat soil is ≤0.5 mm, and the particle size of limestone powder is ≤0.5 mm.

[0010] The preparation method of the above-mentioned Boletus rhodoxanthus cultivation substrate includes the following steps:

[0011] a. Prepare raw materials according to the above-mentioned Boletus rhodoxanthus cultivation substrate;

[0012] b. Add sufficient water to pre-wet fresh sawdust and / or corncobs and chaff at room temperature, cover with a film and stack for 24 h, then add peat soil and silkworm excrement and mix well. Control the moisture content to be 60-65%, and then add the EM bacterial agent and cover with a plastic film to ferment for 20-25 days to obtain a fermented product;

[0013] c. Soak the fresh grains at room temperature for 45-50 h, then take them out and drain, and then mix them with at least one of fresh oats, fresh corn kernels and fresh wheat grains, soil, limestone powder, and the fermented product obtained in step b to obtain a mixture;

[0014] d. Load the mixture obtained in step c into polypropylene mushroom bags, take them out after autoclaving at 120 - 125 °C for 85 - 95 min. Completely dissolve FeSO4, MgSO4, MnSO4 and thermotolerant α - amylase, then spray them on the sterilized mixture and stir evenly. Control the moisture content to be 60 - 65%, then load it into polypropylene plastic bags, take them out after autoclaving at 100 - 110 °C for 28 - 32 min, and place them in a sterile inoculation room to cool to room temperature to obtain the cultivation substrate for Boletus rhodoxanthus.

[0015] Furthermore, in the above - mentioned step c, the soaking time is 48 h.

[0016] Furthermore, in the above - mentioned step d, the specification of the polypropylene mushroom bags for the first sterilization is 25 cm × 45 cm × 0.05 mm, and autoclave at 121 °C for 90 min.

[0017] Furthermore, in the above - mentioned step d, dissolve them completely and mix evenly according to the mass ratio of the total mass of FeSO4, MgSO4, MnSO4 and thermotolerant α - amylase: water = 1:10.

[0018] Furthermore, in the above - mentioned step d, the specification of the polypropylene plastic bags for the second sterilization is 15 cm × 30 cm × 0.05 mm, and autoclave at 105 °C for 30 min.

[0019] The cultivation method of the above - mentioned Boletus rhodoxanthus is as follows: Select the part where the mycelium growth on the edge of the petri dish is relatively neat. Take a 1 cm × 1 cm mycelium block and inoculate it into the cultivation substrate of Boletus rhodoxanthus, and cultivate it at 25 °C. When the mycelium grows to 1 / 3 of the mushroom bag, lower the temperature to 23 °C and cultivate until the bag is full, then cover the soil to produce mushrooms.

[0020] The beneficial effects of the present invention are as follows: The present invention addresses the problems of high contamination rate of artificial cultivation mushroom bags of Boletellus rubrovenulosus, low fruiting rate, and unfavorable large-scale cultivation. In the specially designed cultivation substrate of Boletellus rubrovenulosus, at least one of fresh oats, corn kernels, and wheat grains, fresh grains, and soil are selected to provide the starch and protein required by Boletellus rubrovenulosus, that is, rich carbon sources and nitrogen sources are provided; the fermented rice husks, fresh wood chips, and / or corncobs and peat soil can not only provide humic acid to promote the growth of mycelium, but also change the porosity of the substrate; silkworm excrement can not only provide the organic matter and vitamins required during the growth of mycelium, but also improve the antioxidant properties of the mycelium and enhance the vitality of the mycelium; MnSO4 and MgSO4 can provide the trace elements required during the growth of mycelium, the addition of limestone powder can adjust the stable pH value for the growth of mycelium, reduce the contamination of miscellaneous bacteria, provide Ca ions required for the growth of mycelium, adjust the dry humidity of the substrate, and increase the air permeability of the soil in the substrate; a small amount of FeSO4 inhibits the secretion of mycelium, promotes the kinking of mycelium, helps the formation of rhizomorphs, and enables the mycelium to transform from vegetative growth to reproductive growth; the addition of EM bacterial agents can soften the cellulose in wood chips and rice husks, reduce the tannin content in wood chips, and lower the pH value in silkworm excrement; peat soil contains a large amount of humic acid, which helps the growth of mycelium; the addition of α-amylase can decompose the culture medium, cracking natural polymers such as polysaccharides, proteins, and nucleic acids into small molecule substances that are convenient for the absorption of edible mushroom mycelium and fruiting bodies, and providing nutrients for the growth of edible mushroom mycelium, the formation of primordia, and the growth and development of fruiting bodies.

[0021] The cultivation substrate of Boletellus rubrovenulosus prepared by the method of the present invention is used for the cultivation of Boletellus rubrovenulosus, which can improve the growth rate of mycelium, shorten the growth cycle of mycelium, reduce the contamination rate of mushroom bags, increase the fruiting rate, achieve large-scale cultivation, and bring greater economic and social benefits. Specific embodiments

[0022] The technical solution of the present invention can be specifically implemented in the following manner.

[0023] The cultivation substrate of Boletellus rubrovenulosus, the composition components of which are by mass percentage: at least one of fresh oats, fresh corn kernels, and fresh wheat grains 3 - 8%, fresh grains 20 - 35%, soil 20 - 30%, fresh wood chips and / or corncobs 15 - 25%, silkworm excrement 1 - 5%, rice husks 8 - 15%, FeSO4 0.005 - 0.015%, MgSO4 0.05 - 0.12%, MnSO4 0.01 - 0.03%, peat soil 3 - 7%, limestone powder 0.5 - 2%, EM bacterial agent 0.02 - 0.1%, high-temperature resistant α-amylase 0.02 - 0.08%.

[0024] Preferably, at least one of fresh oats, fresh corn kernels, and fresh wheat grains accounts for 4-6%, fresh grains account for 25-32%, soil accounts for 23-28%, fresh sawdust and / or corncobs account for 18-22%, silkworm excrement accounts for 3-5%, chaff accounts for 10-12%, FeSO4 accounts for 0.008-0.012%, MgSO4 accounts for 0.08-0.12%, MnSO4 accounts for 0.01-0.02%, peat soil accounts for 4-6%, limestone powder accounts for 0.8-1.5%, EM bacterial agent accounts for 0.02-0.05%, and high-temperature resistant α-amylase accounts for 0.04-0.06%.

[0025] More preferably, at least one of fresh oats, fresh corn kernels, and fresh wheat grains accounts for 5%, fresh grains account for 30%, soil accounts for 25%, fresh sawdust and / or corncobs account for 20%, silkworm excrement accounts for 3.8%, chaff accounts for 10%, FeSO4 accounts for 0.01%, MgSO4 accounts for 0.1%, MnSO4 accounts for 0.01%, peat soil accounts for 5%, limestone powder accounts for 1%, EM bacterial agent accounts for 0.03%, and high-temperature resistant α-amylase accounts for 0.05%.

[0026] The above-mentioned fresh grains are fresh millet.

[0027] The particle size of the above-mentioned soil is 0.1-1 cm, the particle size of fresh sawdust and / or corncobs is 0.5-2 cm, the particle size of peat soil is ≤0.5 mm, and the particle size of limestone powder is ≤0.5 mm.

[0028] The preparation method of the above-mentioned Boletus rhodoxanthus cultivation substrate includes the following steps:

[0029] 1. Weigh the ingredients according to the above-mentioned Boletus rhodoxanthus cultivation substrate formula. Then, pre-wet the fresh sawdust and / or corncobs and chaff to make the culture material fully absorb water. Next, add the silkworm excrement, peat soil, and EM bacterial agent to the culture material and adjust the water content to 60-65%. Build a pile for fermentation, with a pile height of 1-1.2 m, a pile width of 0.8-1.2 m, and an unlimited length. After building the pile, cover it with a plastic film to keep it moist. When the temperature rises above 65°C, maintain it for 48 h and then turn the pile once. When turning the material, turn it up and down and inside and outside to ensure uniform fermentation. Turn the pile 3 times in this way and ferment for 20-25 days to obtain the fermented material. The fermented culture material is brownish-brown and has no pungent smell;

[0030] 2. Weigh at least one of fresh oats, fresh corn kernels, and fresh wheat grains, and soak the fresh grains in water for 45-50 h respectively to make them fully absorb water and have no white core;

[0031] 3. Take out and drain at least one of the soaked fresh oats, fresh corn kernels, and fresh wheat grains, and mix the fresh grains with the fermentation material. Add soil and lime powder and stir evenly. Control the moisture content to be 60 - 65%, and bag it (polypropylene mushroom bags, with specifications of 25 cm × 45 cm × 0.05 mm) and sterilize at 120 - 125 °C for 85 - 95 min, then take it out and cool down.

[0032] 4. Mix FeSO4, MgSO4, MnSO4, and heat-resistant α-amylase according to the ratio with water and then add it to the sterilized culture material, stir evenly. Control the moisture content of the culture material to be 60 - 65%, and bag it (polypropylene plastic mushroom bags, with specifications of 15 cm × 30 cm × 0.05 mm) or bottle it (1100 ml) and sterilize at 100 - 110 °C for 28 - 32 min. After the sterilization is completed, put it into a sterile inoculation room and cool it to room temperature. The medium is made, and the cultivation substrate for Boletus rhodopunctatus is obtained.

[0033] Mixing and fermenting fresh sawdust and / or corn cobs, rice hulls with silkworm excrement, peat soil, and EM bacterial agents can soften the fermentation material, effectively reduce the number of miscellaneous bacteria in the culture material, increase the number of beneficial bacteria, and is conducive to the absorption of Boletus mycelium. Using secondary sterilization results in less nutrient loss of the culture material compared to primary sterilization, and the humidity of the culture material is more uniform, which is beneficial to the germination and uniform growth of the mycelium. Reducing the temperature and shortening the time during the second sterilization can effectively maintain the activity of heat-resistant α-amylase.

[0034] The cultivation method of the above-mentioned Boletus rhodopunctatus is as follows: Select the part where the mycelium growth on the edge of the petri dish is relatively neat in the culture dish, take a 1 cm × 1 cm piece of the strain and inoculate it into the cultivation substrate of Boletus rhodopunctatus, and culture it at 25 °C. When the mycelium grows to 1 / 3 of the mushroom bag, lower the temperature to 23 °C and culture until the bag is full and then cover the soil to produce mushrooms.

[0035] The technical solutions and effects of the present invention will be further described through actual examples below.

[0036] Example

[0037] (1) The present invention provides 5 groups of examples of cultivating Boletus rhodopunctatus using the cultivation substrate of the present invention (Examples 1 - 5), and 1 group of conventional cultivation substrate (Comparative Example 1). The specific components of the cultivation substrates of Boletus rhodopunctatus in Examples 1 - 5 and the cultivation substrate of Comparative Example 1 are shown in Table 1. Among them, the EM bacterial agent is purchased from Shanxi Changtai Animal Pharmaceutical Co., Ltd., and the implementation standard is: Q / SCY010 - 213.

[0038] Table 1 Mass percentage of the components of the cultivation substrates in Examples 1 - 5 and Comparative Example 1

[0039]

[0040]

[0041] According to the cultivation method of Boletus rhodopunctus of the present invention: Select the part where the mycelium grows neatly at the edge of the petri dish in the culture dish, take a 1 cm × 1 cm mycelium block and inoculate it into the cultivation substrate, cultivate at 25 °C, and when the mycelium grows to 1 / 3 of the mushroom bag, lower the temperature to 23 °C and cultivate until the bag is full and cover with soil to produce mushrooms. The cultivation effects of Examples 1-5 and Comparative Example 1 are shown in Table 2.

[0042] Table 2 Cultivation effects of Examples 1-5 and Comparative Example 1

[0043] Culture medium Feeding time Full bag time Contamination rate Mushroom fruiting rate Example 1 2 days 54 days 8.52％ 93.45％ Example 2 2 days 53 days 8.35％ 92.82％ Example 3 2 days 55 days 7.45％ 91.47％ Example 4 2 days 58 days 9.70％ 90.20％ Example 5 2 days 50 days 5.20％ 96.38％ Comparative example 1 3 days 63 days 15.35％ 80.50％

[0044] It can be seen from Examples 1-5 and Comparative Example 1 that by using the cultivation substrate components of Boletus rhodopunctus of the present invention, the growth cycle of the mycelium can be effectively shortened to less than 58 days, the pollution rate of the mushroom bag can be reduced to less than 10%, and the mushroom emergence rate of Boletus rhodopunctus can be increased to more than 90%.

[0045] (2) The present invention provides a group of formulations of the components of the cultivation substrate of the present invention and a cultivation substrate of Boletus rhodopunctus (Example 5) prepared according to the preparation method of the present invention; a group of cultivation substrates according to the conventional cultivation substrate formula and preparation method (Comparative Example 1); and 5 groups of cultivation substrates that do not completely adopt the component formula and preparation method of the cultivation substrate of the present invention (Comparative Examples 2-6) for the cultivation of Boletus rhodopunctus. The specific components of the cultivation substrates of Example 5 and Comparative Examples 1-6 are shown in Table 3.

[0046] Table 3 Mass percentages of cultivation substrate components

[0047]

[0048]

[0049] Among them, Example 5 was prepared completely according to the preparation method of the cultivation substrate of Boletus rhodopunctus of the present invention; Comparative Example 1 was prepared according to the conventional cultivation substrate method; Comparative Examples 2, 3, and 5 were sterilized at high temperature for 2 hours once, and Comparative Examples 4 and 6 were sterilized at high temperature for 2 hours twice, and the remaining steps were prepared according to the preparation method of the present invention.

[0050] Using the cultivation substrates prepared above, according to the cultivation method of Boletus rhodopunctus of the present invention: Select the part where the mycelium grows neatly at the edge of the petri dish in the culture dish, take a 1 cm × 1 cm mycelium block and inoculate it into the cultivation substrate, cultivate at 25 °C, and when the mycelium grows to 1 / 3 of the mushroom bag, lower the temperature to 23 °C and cultivate until the bag is full and cover with soil to produce mushrooms. The cultivation effects of Example 5 and Comparative Examples 1-6 are shown in Table 4.

[0051] Table 4 Cultivation effects of Example 5 and Comparative Examples 1-6

[0052] Culture medium Feeding time Full bag time Contamination rate Mushroom fruiting rate Example 5 2 days 50 days 5.20％ 96.38％ Comparative example 1 3 days 65 days 15.35％ 80.50％ Comparative example 2 2 days 60 days 7.50％ 89.55％ Comparative example 3 2 days 59 days 8.25％ 90.60％ Comparative example 4 2 days 52 days 6.56％ 94.15％ Comparative example 5 2 days 58 days 7.20％ 90.95％ Comparative example 6 2 days 54 days 6.15％ 92.38％

[0053] As can be seen from Example 5 and Comparative Examples 1-6, by using the cultivation substrate for Boletus rhodoxanthus of the present invention and its preparation method, the growth cycle of the mycelium can be effectively shortened, the contamination rate of the mushroom bags can be reduced, and the fruiting rate of Boletus rhodoxanthus can be increased.

Claims

1. Cultivation substrate for Boletus rubrovenulosus, characterized in that, The composition by mass percentage is as follows: at least one of fresh oats, fresh corn kernels and fresh wheat grains is 3 - 8%, fresh grains are 20 - 35%, soil is 20 - 30%, fresh sawdust and / or corncobs are 15 - 25%, silkworm excrement is 1 - 5%, rice husks are 8 - 15%, FeSO4 is 0.005 - 0.015%, MgSO4 is 0.05 - 0.12%, MnSO4 is 0.01 - 0.03%, peat soil is 3 - 7%, limestone powder is 0.5 - 2%, EM microbial agent is 0.02 - 0.1%, and heat-resistant α-amylase is 0.02 - 0.08%. The preparation method of the above-mentioned Boletellus rubrovenulosus cultivation substrate comprises the following steps: a. Prepare raw materials according to the composition of the Boletellus rubrovenulosus cultivation substrate; b. Add sufficient water to pre-wet fresh sawdust and / or corncobs and rice husks at room temperature, then cover with a film and stack for 24 h. After that, add peat soil and silkworm excrement and mix evenly. Control the water content to be 60 - 65%, and then add the EM microbial agent and cover with a plastic film to ferment for 20 - 25 days to obtain a fermented product; c. Soak fresh grains at room temperature for 45 - 50 h, then take them out and drain. Then mix them with at least one of fresh oats, fresh corn kernels and fresh wheat grains, soil, limestone powder, and the fermented product obtained in step b to obtain a mixture; d. Pack the mixture obtained in step c into polypropylene mushroom bags, sterilize at 120 - 125 °C for 85 - 95 min and then take out. Completely dissolve FeSO4, MgSO4, MnSO4 and heat-resistant α-amylase and spray them on the sterilized mixture and stir evenly. Control the water content to be 60 - 65%, then pack them into polypropylene plastic bags, sterilize at 100 - 110 °C for 28 - 32 min and then take out, and put them into a sterile inoculation room to cool to room temperature to obtain the Boletellus rubrovenulosus cultivation substrate.

2. The cultivation substrate of Boletellus rubrovenulosus according to claim 1, characterized in that, The composition by mass percentage is as follows: at least one of fresh oats, fresh corn kernels and fresh wheat grains is 4 - 6%, fresh grains are 25 - 32%, soil is 23 - 28%, fresh sawdust and / or corncobs are 18 - 22%, silkworm excrement is 3 - 5%, rice husks are 10 - 12%, FeSO4 is 0.008 - 0.012%, MgSO4 is 0.08 - 0.12%, MnSO4 is 0.01 - 0.02%, peat soil is 4 - 6%, limestone powder is 0.8 - 1.5%, EM microbial agent is 0.02 - 0.05%, and heat-resistant α-amylase is 0.04 - 0.06%.

3. The cultivation substrate of Boletus rhodopetersii according to claim 2, characterized in that, The composition by mass percentage is as follows: at least one of fresh oats, fresh corn kernels and fresh wheat grains is 5%, fresh grains are 30%, soil is 25%, fresh sawdust and / or corncobs are 20%, silkworm excrement is 3.8%, rice husks are 10%, FeSO4 is 0.01%, MgSO4 is 0.1%, MnSO4 is 0.01%, peat soil is 5%, limestone powder is 1%, EM microbial agent is 0.03%, and heat-resistant α-amylase is 0.05%.

4. The cultivation substrate of Boletus rhodoxanthus according to any one of claims 1-3, characterized in that: The particle size of the soil is 0.1 - 1 cm, the particle size of the fresh sawdust and / or corncobs is 0.5 - 2 cm, the particle size of the peat soil is ≤0.5 mm, and the particle size of the limestone powder is ≤0.5 mm.

5. The cultivation substrate of Boletus rhodopetersii according to claim 1, characterized in that: In step c, the soaking time is 48 h.

6. The cultivation substrate of Boletus rhodopetersii according to claim 1, characterized in that: In step d, the polypropylene spawn bags have a specification of 25 cm × 45 cm × 0.05 mm and are autoclaved at 121 °C for 90 min; the polypropylene plastic bags have a specification of 15 cm × 30 cm × 0.05 mm and are autoclaved at 105 °C for 30 min.

7. The cultivation substrate of Boletus rhodopetersii according to claim 1, characterized in that: In step d, FeSO4, MgSO4, MnSO4 and thermotolerant α-amylase are completely dissolved and mixed evenly according to the mass ratio of the total mass of FeSO4, MgSO4, MnSO4 and thermotolerant α-amylase to water = 1:

10.

8. Cultivation method of Leccinum roseonervosum, characterized in that: Select the part where the mycelium growth on the edge of the petri dish is relatively neat, take a 1 cm × 1 cm mycelium block and inoculate it into the Boletus rhodopetersii cultivation substrate according to any one of claims 1-7, and culture it at 25 °C. When the mycelium grows to 1 / 3 of the spawn bag, reduce the temperature to 23 °C and culture until the bag is full and then cover with soil to produce mushrooms.

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