Flammulina velutipes culture medium taking broken shells as raw material and preparation method thereof
The method for preparing enoki mushroom culture medium by compounding crushed shells, differentiated pretreatment, staged fermentation, and compound nutrient components has solved the problems of scarce sawdust raw materials and poor culture medium performance, and achieved efficient and stable growth and high yield of enoki mushrooms, with significant economic and environmental benefits.
Patent Information
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- HEFEI FUQUAN MODERN AGRI SCI & TECH
- Filing Date
- 2026-03-19
- Publication Date
- 2026-05-15
AI Technical Summary
Current methods for cultivating enoki mushrooms include the scarcity of sawdust raw materials, high procurement costs, a single nutrient structure in the culture medium, a high risk of contamination by miscellaneous microorganisms, and a long mycelium growth cycle. Furthermore, existing methods of utilizing crushed shells cannot meet the diverse nutritional needs of enoki mushroom growth. Insufficient pretreatment of crushed shells and inadequate fermentation processes result in poor culture medium stability, making it difficult to achieve large-scale application.
By using a combination of crushed nut shells, crushed livestock and poultry shells, and crushed fruit and vegetable shells, a differentiated pretreatment process is designed. This is combined with staged fermentation and compound fermentation agents, along with compound nutrient components such as kudzu root powder and distiller's grains. The raw material ratio is optimized to construct a balanced nutrient system. Through a sterilization process that combines high pressure and normal pressure, the environment for mycelium growth and fruiting is controlled to achieve stable growth of enoki mushrooms.
It improves mycelial growth rate, shortens the mycelial growth cycle, reduces contamination rate by miscellaneous bacteria, increases mushroom yield and product quality, realizes the efficient resource utilization of agricultural waste, reduces the cost of culture medium preparation, and has good economic and environmental benefits.
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Abstract
Description
Technical Field
[0001] This invention relates to the field of edible fungi cultivation technology, specifically to a enoki mushroom culture medium using crushed shells as raw material and its preparation method. Background Technology
[0002] As a common edible and medicinal fungus, enoki mushrooms are experiencing a continuous increase in market demand. The formulation and preparation process of the culture medium for its artificial cultivation directly affect cultivation efficiency and product quality. Currently, sawdust is the main raw material for enoki mushroom cultivation. However, with the increasing protection of forestry resources, sawdust is becoming increasingly scarce, and procurement costs are constantly rising. At the same time, traditional sawdust culture media suffer from a single nutrient structure, which can easily lead to slow mycelial growth, a long mycelial growth cycle, and a high risk of contamination by other fungi, ultimately affecting the yield and nutritional quality of the mushrooms.
[0003] To alleviate the raw material shortage, existing technologies attempt to apply agricultural waste shells to the preparation of enoki mushroom culture media, but these technologies still have many shortcomings. Current methods for utilizing shells mostly employ single-species shells, failing to meet the diverse nutritional needs of enoki mushroom growth; pretreatment methods are relatively simple, lacking differentiation based on the characteristics of different varieties, resulting in the ineffective removal of undesirable components that affect mycelial absorption and utilization; fermentation processes are mostly based on a single temperature, leading to insufficient decomposition and poor culture medium stability; furthermore, there are issues with unreasonable shell ratio design and a single nutrient system, lacking the inclusion of compound nutrient components, resulting in poor culture medium performance, low efficiency in shell resource utilization, and difficulty in achieving large-scale promotion and application. Summary of the Invention
[0004] The primary objective of this invention is to provide a culture medium for enoki mushrooms using crushed shells as raw material and a method for preparing the same.
[0005] A further objective of this invention is to provide a culture medium for enoki mushrooms using crushed shells as raw material. The raw material, by weight, comprises: 5 to 30 parts of crushed nut shells, 3 to 15 parts of crushed livestock and poultry shells, 2 to 10 parts of crushed fruit and vegetable shells, 4 to 16 parts of wheat bran, 2 to 6 parts of corn flour, 0.8 to 2 parts of gypsum powder, 0.8 to 2 parts of sucrose, 0.4 to 1.0 parts of superphosphate, 0.08 to 0.25 parts of magnesium sulfate, 0.1 to 0.5 parts of urea, 0.5 to 2 parts of kudzu root powder, 2 to 5 parts of distiller's grains, 3 to 5 parts of soybean cake, and an appropriate amount of water.
[0006] Preferably, the nut shells are one or more of walnut shells, peanut shells, and chestnut shells; the poultry shells are one or more of chicken shells, duck shells, and goose shells; and the fruit and vegetable shells are one or more of citrus shells, apple shells, and pear shells.
[0007] Preferably, the particle size of the crushed nut shells, livestock and poultry shells, and fruit and vegetable shells is 0.5 to 10 mm.
[0008] The method for preparing the enoki mushroom culture medium using crushed shells as raw material includes the following steps: (1) Shell pretreatment: crush the shells of nuts, livestock and poultry, and fruits and vegetables to a particle size of 0.5 to 10 mm. Different types of shells are subjected to differentiated warm soaking or additive treatment. After draining to a moisture content of 48 to 57%, they are mixed evenly and set aside. (2) Mixing ingredients: Crush soybean cake, and then add the pre-treated mixed crushed shells, wheat bran, corn flour, gypsum powder, sucrose, superphosphate, magnesium sulfate, urea, kudzu root powder, distiller's grains, and crushed soybean cake to the mixing equipment in sequence. During the mixing process, add water to adjust the moisture content of the mixture to 59 to 67%, and stir until the material is uniform and free of lumps. (3) Fermentation treatment: The mixture is piled into a pile with a height of 80 to 90 cm and a width of 100 to 110 cm, covered with plastic film for staged fermentation. During the fermentation process, the pile is turned frequently and water is added to maintain the moisture content. Fermentation agents can be added. Fermentation continues until the pile has no odor, and fermented material is obtained. (4) Sterilization and inoculation: The fermentation material is put into a polypropylene plastic bag, the filling amount is 70% of the bag volume, and after sterilization, it is cooled to below 30°C. Inoculate the enoki mushroom spawn in a sterile environment with a colony count ≤10cfu / cm². (5) Mycelium growth and cultivation: Place the inoculated bags in the cultivation room, control the temperature, relative humidity, light intensity and ventilation conditions of the cultivation room, and cultivate until the mycelium has fully grown into the bags and is fully mature; (6) Mushroom management: Transfer mature mushroom bags to the mushroom growing room, control the temperature, relative humidity, light intensity and ventilation conditions of the mushroom growing room, and harvest when the enoki mushrooms grow to a stipe length of 14 to 23 cm and a cap diameter of 0.7 to 1.5 cm.
[0009] Preferably, in step (1), nut shells are soaked in 50°C warm water for 12 to 36 hours, with the water changed twice, and drained until the moisture content is 48 to 57%; livestock and poultry shells are soaked in 60°C warm water for 8 to 12 hours, with 0.2 to 0.5% edible alkali added, and drained until the moisture content is 48 to 57%; fruit and vegetable shells are used directly or soaked in 0.2% citric acid for 6 hours and then drained until the moisture content is 48 to 57%.
[0010] Preferably, in step (2), the stirring is either a single low-speed stirring or a combination of high-speed stirring and low-speed stirring, with a stirring speed of 150 to 400 r / min, a total stirring time of 15 to 30 minutes, and the moisture content of the mixture is adjusted to 59 to 67%; when high-speed stirring and low-speed stirring are combined, high-speed stirring is performed for 10 to 15 minutes first, followed by low-speed stirring for 15 minutes.
[0011] Preferably, in step (3), the height of the material pile is 80 to 90 cm and the width is 100 to 110 cm; the temperature of the first stage of the phased fermentation is 28 to 36°C, the fermentation lasts for 3 to 4 days, and the pile is turned over 1 to 2 times a day; the temperature of the second stage is 24 to 30°C, the fermentation lasts for 4 to 6 days, and the pile is turned over once every 2 days; the fermentation agent is a compound agent composed of Bacillus subtilis and yeast in a mass ratio of 1:1, and its addition amount is 0.08 to 0.2% of the mass of the mixture.
[0012] Preferably, in step (4), the sterilization process is high pressure sterilization, normal pressure sterilization, or a combination of high pressure and normal pressure sterilization; the high pressure sterilization temperature is 121°C and the sterilization time is 1 to 2.5 hours, the normal pressure sterilization temperature is 100°C and the sterilization time is 12 to 16 hours; the inoculation amount of enoki mushroom spawn is 4 to 8% of the mass of the fermentation material.
[0013] Preferably, in step (5), the temperature of the cultivation room is 21 to 26°C, the relative humidity is 65 to 72%, the light intensity is ≤50 lux, the room is ventilated 1 to 2 times a day for 15 to 25 minutes each time, and the cultivation time is 25 to 38 days; in step (6), the temperature of the fruiting room is 7 to 13°C, the relative humidity is 85 to 94%, the light intensity is 50 to 100 lux, the room is ventilated 1 to 2 times a day for 10 to 25 minutes each time; the harvesting specifications of enoki mushrooms are a stem length of 14 to 23 cm and a cap diameter of 0.7 to 1.5 cm.
[0014] Compared with the prior art, the beneficial effects of the present invention are: 1. This invention provides a enoki mushroom culture medium using crushed shells as raw material and its preparation method, which effectively solves the problems of raw material scarcity, unreasonable process and poor performance of existing enoki mushroom cultivation media.
[0015] 2. This invention uses a mixture of nut shells, livestock and poultry shells, and fruit and vegetable shells as the main raw materials, making full use of various agricultural waste materials to achieve efficient resource utilization of agricultural waste. This not only alleviates the shortage of traditional sawdust raw materials, but also reduces the preparation cost of culture medium, thus achieving both good economic and environmental benefits.
[0016] 3. Differentiated pretreatment processes were designed for different types of crushed shells. Combined with staged fermentation processes and the application of compound fermentation agents, the degree of decomposition of crushed shells was effectively improved, components that are not conducive to mycelial growth were removed from the crushed shells, the physicochemical properties of the culture medium were optimized, the possibility of contamination by other microorganisms was reduced, and the stability and applicability of the culture medium were improved.
[0017] 4. At the same time, by rationally designing the raw material ratio and combining it with compound nutrient components such as kudzu root powder, distiller's grains, and soybean cake, a balanced nutritional system is constructed to meet the nutritional needs of enoki mushrooms at different growth stages, optimize mycelial growth, and ensure the continuity and stability of fruiting. Detailed Implementation
[0018] The technical solutions in the embodiments of the present invention will be clearly and completely described below. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.
[0019] Example 1:
[0020] The raw material components, by weight, are: 10 parts crushed nut shells, 5 parts wheat bran, 3 parts corn flour, 1 part gypsum powder, 1 part sucrose, 0.5 parts superphosphate, 0.1 parts magnesium sulfate, 0.2 parts urea, and appropriate amount of water. Walnut shells are selected as the crushed nut shells and crushed to a particle size of 1-8mm.
[0021] Preparation method: Shell pretreatment: Crush the walnut shells to a particle size of 1-8mm, soak them in 50℃ warm water for 24 hours, changing the water twice during the process, then drain them until the moisture content is 50-55%, and set aside.
[0022] Mixing ingredients: Add the pretreated walnut shells, wheat bran, corn flour, gypsum powder, sucrose, superphosphate, magnesium sulfate, and urea into the mixing equipment in sequence, and mix at low speed for 15 minutes at a mixing speed of 150 r / min. During the mixing process, slowly add water to adjust the moisture content of the mixture to 60-65% and mix until the material is uniform and free of lumps.
[0023] Fermentation treatment: Pile the mixture into a pile 80cm high and 100cm wide, cover it with plastic film and let it ferment naturally. The initial fermentation temperature is controlled at 25-30℃. During the fermentation process, turn the pile once a day and add an appropriate amount of water to maintain the moisture content. Ferment for 7 days until the pile has no odor and turns dark brown, and you will get the fermented material.
[0024] Sterilization and inoculation: The fermentation material is packed into polypropylene plastic bags, filling each bag to 70% of its volume. Autoclaving is performed at 121°C for 2 hours. After sterilization, the mixture is cooled to below 30°C. Inoculation with *Flammulina velutipes* spawn is then carried out in a sterile environment, with an inoculation amount of 5% of the fermentation material mass. The aseptic colony count should be ≤10 CFU / cm³. 2 The enoki mushroom strain used is a commercially available conventional enoki mushroom cultivation strain.
[0025] Mycelium growth and cultivation: Place the inoculated bags into the cultivation room, control the temperature of the cultivation room to be 22-25℃, the relative humidity of the air to be 65-70%, maintain a dark environment, and the light intensity to be ≤50 lux. Ventilate once a day for 15 minutes each time. Cultivate for 30-35 days until the mycelium has fully grown into the bags and is fully mature, and then transfer to the fruiting management.
[0026] Mushroom management: Transfer mature mushroom bags to the mushroom growing room, control the temperature of the growing room at 10-12℃ and the relative humidity at 85-90%, and hang a 15W light bulb every 3-5m to maintain weak light to induce stipe elongation. The light intensity is 50-100 lux. Ventilate once a day for 10-20 minutes each time. Harvest when the stipe grows to 15-20cm and the cap diameter is 0.8-1.2cm.
[0027] Example 2:
[0028] Based on the technical solution of Example 1, the types of crushed shells are increased and the raw material ratio is adjusted to improve the nutritional balance of the culture medium. The raw material components by weight are: 15 parts nut crushed shells, 5 parts livestock and poultry crushed shells, 8 parts wheat bran, 4 parts corn flour, 1.2 parts gypsum powder, 1.2 parts sucrose, 0.6 parts superphosphate, 0.15 parts magnesium sulfate, 0.3 parts urea, 1 part kudzu root powder, and appropriate amount of water. Peanut shells are used for nut crushing, and chicken shells are used for livestock and poultry crushing, both crushed to a particle size of 1-8 mm.
[0029] Preparation method: Shell pretreatment: Crush peanut shells and chicken shells separately to a particle size of 1-8mm. Soak peanut shells in 50℃ warm water for 24 hours, changing the water twice during this period. Drain them until the moisture content is 50-55%. Soak chicken shells in 60℃ warm water for 12 hours, add 0.3% edible alkali, drain them until the moisture content is 50-55%. Mix the two types of crushed shells evenly and set aside.
[0030] Mixing ingredients: Add the pretreated mixed crushed shells, wheat bran, corn flour, gypsum powder, sucrose, superphosphate, magnesium sulfate, urea, and kudzu root powder to the mixing equipment in sequence, and mix at low speed for 20 minutes at a mixing speed of 180 r / min. Add water slowly during the mixing process to adjust the moisture content of the mixture to 60-65%, and mix until the material is uniform and free of lumps.
[0031] Fermentation treatment: Pile the mixture into a pile 90cm high and 110cm wide, cover it with plastic film for fermentation, and control the initial fermentation temperature at 28-32℃. Turn the pile once a day for the first 3 days of fermentation, and turn it once every 2 days in the later stage. Add an appropriate amount of water to maintain the moisture content. Ferment for 8 days until the pile has no odor and the mycelium has initially germinated, and the fermented material is obtained.
[0032] Sterilization and inoculation: Pack the fermentation material into polypropylene plastic bags, filling each bag to 70% of its volume. Sterilize using normal pressure at 100℃ for 14-16 hours. After sterilization, cool to below 30℃ and inoculate with *Flammulina velutipes* spawn in a sterile environment. The inoculation amount is 6% of the fermentation material mass. The aseptic colony count should be ≤10 CFU / cm³. 2 The enoki mushroom strain used is a commercially available conventional enoki mushroom cultivation strain.
[0033] Mycelium growth and cultivation: Place the inoculated bags into the cultivation room, control the temperature of the cultivation room at 23-26℃, the relative humidity of the air at 68-72%, maintain a dark environment with a light intensity of ≤50 lux, ventilate twice a day for 15 minutes each time, and cultivate for 32-37 days until the mycelium has fully grown into the bags and is fully mature, then transfer to the fruiting management.
[0034] Mushroom management: Transfer mature mushroom bags to the mushroom growing room, control the temperature of the growing room at 9-13℃ and the relative humidity at 86-92%, and hang a 15W light bulb every 3-5m to maintain weak light to induce stipe elongation. The light intensity is 50-100 lux. Ventilate once a day for 15-25 minutes each time. Harvest when the stipe reaches 16-21cm in length and the cap diameter is 0.9-1.3cm.
[0035] Example 3:
[0036] Based on the technical solution of Example 2, the proportion of crushed shells is further increased, auxiliary nutrient components are added, and the fermentation process is optimized. The raw material components by weight are: 20 parts nut crushed shells, 10 parts livestock and poultry crushed shells, 5 parts fruit and vegetable crushed shells, 12 parts wheat bran, 5 parts corn flour, 1.5 parts gypsum powder, 1.5 parts sucrose, 0.8 parts superphosphate, 0.2 parts magnesium sulfate, 0.4 parts urea, 1.5 parts kudzu root powder, 3 parts distiller's grains, and appropriate amount of water. Chestnut shells are used for nut crushed shells, duck shells are used for livestock and poultry crushed shells, and citrus shells are used for fruit and vegetable crushed shells. All are crushed to a particle size of 1-8 mm.
[0037] Preparation method: Shell pretreatment: Crush chestnut shells, duck shells, and citrus shells into particles of 1-8mm. Soak chestnut shells in 50℃ warm water for 24 hours, changing the water twice during this period. Drain them until the moisture content is 50-55%. Soak duck shells in 60℃ warm water for 12 hours, adding 0.3% edible alkali. Drain them until the moisture content is 50-55%. Citrus shells can be used directly. Mix the three types of crushed shells evenly and set aside.
[0038] Mixing ingredients: Add the pretreated mixed crushed shells, wheat bran, corn flour, gypsum powder, sucrose, superphosphate, magnesium sulfate, urea, kudzu root powder, and distiller's grains into the mixing equipment in sequence. First, mix at high speed for 10 minutes at 400 r / min, then mix at low speed for 15 minutes at 150 r / min. Add water in stages during the mixing process to adjust the moisture content of the mixture to 62-66%. Mix until the material is uniform and free of lumps.
[0039] Fermentation treatment: The mixed material is piled into a heap 90cm high and 110cm wide, covered with plastic film, and fermented in stages. The first stage of fermentation is controlled at 30-35℃ for 4 days, with the heap turned once a day. The second stage of fermentation is controlled at 25-30℃ for 5 days, with the heap turned once every 2 days. During the fermentation process, 0.1% of fermentation agent is added. The fermentation agent is a compound agent of Bacillus subtilis and yeast in a 1:1 mass ratio. An appropriate amount of water is added to maintain the moisture content. Fermentation continues until the heap has no odor, turns dark brown, and has a faint fermented aroma, thus obtaining the fermented material.
[0040] Sterilization and Inoculation: Pack the fermentation material into polypropylene plastic bags, filling each bag to 70% of its volume. Use a combination of high-pressure and normal-pressure sterilization: first, autoclave at 121℃ for 1 hour, then sterilize at 100℃ under normal pressure for 8 hours. After sterilization, cool to below 28℃. Inoculate with *Flammulina velutipes* spawn in a sterile environment, using 7% of the fermentation material's mass. After inoculation, gently shake the bags to ensure even distribution of the spawn. The colony count in the sterile environment should be ≤10 CFU / cm³. 2 The enoki mushroom strain used is a commercially available conventional enoki mushroom cultivation strain.
[0041] Mycelium growth and cultivation: Place the inoculated bags into the cultivation room, control the temperature of the cultivation room at 22-24℃, the relative humidity of the air at 66-70%, maintain a completely dark environment, and the light intensity at ≤10 lux. Ventilate twice a day for 20 minutes each time. When turning the bags, exchange the positions of the bags from top to bottom and inside to outside. Cultivate for 28-33 days until the mycelium has fully grown into the bags and is fully mature, and then transfer to the fruiting management.
[0042] Mushroom management: Transfer mature mushroom bags to the mushroom growing room, control the temperature of the growing room at 8-12℃ and the relative humidity at 87-93%, and install a 15W light bulb every 3-5m to maintain weak light to induce stipe elongation. The light intensity should be 50-100 lux. Ventilate twice a day for 15 minutes each time. Harvest when the stipe reaches 17-22cm in length and the cap diameter reaches 1.0-1.4cm.
[0043] Example 4:
[0044] Based on the technical solution of Example 3, the upper limit of the crushed shell ratio is further expanded, the pretreatment and sterilization process is optimized, and the performance of the culture medium is improved. The raw material components by weight are: 30 parts of nut crushed shells, 15 parts of livestock and poultry crushed shells, 10 parts of fruit and vegetable crushed shells, 16 parts of wheat bran, 6 parts of corn flour, 2 parts of gypsum powder, 2 parts of sucrose, 1.0 part of superphosphate, 0.25 parts of magnesium sulfate, 0.5 parts of urea, 2 parts of kudzu root powder, 5 parts of distiller's grains, 5 parts of soybean cake, and appropriate amount of water. The nut crushed shells are a mixture of walnut shells and peanut shells, the livestock and poultry crushed shells are goose shells, and the fruit and vegetable crushed shells are apple shells, all of which are crushed to a particle size of 1-10 mm.
[0045] Preparation method: Shell pretreatment: Crush walnut shells, peanut shells, goose shells, and apple shells into particles of 1-10mm. Mix walnut shells and peanut shells and soak them in 50℃ warm water for 36 hours, changing the water twice during this period. Drain them until the moisture content is 52-57%. Soak goose shells in 60℃ warm water for 12 hours, add 0.5% edible alkali, and drain them until the moisture content is 52-57%. Soak apple shells in 0.2% citric acid for 6 hours, and drain them until the moisture content is 52-57%. Mix the three types of crushed shells evenly and set aside.
[0046] Mixing ingredients: After crushing the soybean cake, add it to the mixing equipment in sequence with the pre-treated mixed crushed shells, bran, corn flour, gypsum powder, sucrose, superphosphate, magnesium sulfate, urea, kudzu root powder, and distiller's grains. Extend the mixing time to 30 minutes. First, mix at high speed for 15 minutes at 350 r / min, then mix at low speed for 15 minutes at 180 r / min. Use a staged water addition method. First, add 70% of the total water and mix for 15 minutes. Then add the remaining 30% of the water and continue mixing for 15 minutes. Adjust the moisture content of the mixture to 63-67% and mix until the material is uniform and free of lumps.
[0047] Fermentation treatment: The mixed material is piled into a heap 90cm high and 110cm wide, covered with plastic film, and fermented in stages. The first stage fermentation temperature is 32-36℃, fermentation lasts for 3 days, and the heap is turned twice a day. The second stage fermentation temperature is 26-30℃, fermentation lasts for 6 days, and the heap is turned once every 2 days. During the fermentation process, 0.2% of fermentation inoculant is added. The fermentation inoculant is a compound inoculant of Bacillus subtilis and yeast in a mass ratio of 1:1. An appropriate amount of water is added to maintain the moisture content. After fermentation, the fermented material is spread out to cool to room temperature, and impurities are removed to obtain high-quality fermented material.
[0048] Sterilization and inoculation: The fermentation material is packed into polypropylene plastic bags, filling each bag to 70% of its volume. Autoclaving is performed at 121°C for 2.5 hours. After sterilization, the mixture is cooled to below 25°C. Inoculation with *Flammulina velutipes* spawn is then carried out in a sterile environment at an inoculation rate of 8% of the fermentation material's mass. Strict aseptic techniques must be followed during inoculation. The aseptic colony count should be ≤10 CFU / cm³. 2 The enoki mushroom strain used is a commercially available conventional enoki mushroom cultivation strain.
[0049] Mycelium growth and cultivation: Place the inoculated bags into the cultivation room, control the temperature of the cultivation room at 21-23℃, the relative humidity of the air at 65-68%, maintain a completely dark environment, and the light intensity at ≤10 lux. Ventilate twice a day for 25 minutes each time. Regularly check the bags and promptly deal with bags contaminated by other microorganisms. Cultivate for 25-30 days until the mycelium has fully grown into the bags and is fully mature, then transfer to fruiting management.
[0050] Mushroom management: Transfer mature mushroom bags to the mushroom growing room, control the temperature of the growing room at 7-11℃ and the relative humidity at 88-94%, and install a 15W light bulb every 3-5m. Use intermittent low light induction, 3 times a day, 30 minutes each time, with a light intensity of 50-100 lux. Ventilate twice a day for 20 minutes each time to avoid water accumulation in the mushrooms. Harvest when the stipe reaches 18-23cm in length and the cap diameter reaches 1.1-1.5cm.
[0051] Example 5:
[0052] Based on the technical solution of Example 4, the proportion of crushed shells is increased to the lower limit, and the mushroom cultivation management process is optimized to form a complete technical coverage. The raw material components by weight are: 5 parts nut crushed shells, 3 parts livestock and poultry crushed shells, 2 parts fruit and vegetable crushed shells, 4 parts wheat bran, 2 parts corn flour, 0.8 parts gypsum powder, 0.8 parts sucrose, 0.4 parts superphosphate, 0.08 parts magnesium sulfate, 0.1 parts urea, 0.5 parts kudzu root powder, 2 parts distiller's grains, 3 parts soybean cake, and appropriate amount of water. Chestnut shells are used for nut crushed shells, chicken shells are used for livestock and poultry crushed shells, and pear shells are used for fruit and vegetable crushed shells. All are crushed to a particle size of 0.5-8mm.
[0053] Preparation method: Shell pretreatment: Crush chestnut shells, chicken shells, and pear shells into particles of 0.5-8mm. Soak chestnut shells in 50℃ warm water for 12 hours, changing the water twice during this period. Drain until the moisture content is 48-53%. Soak chicken shells in 60℃ warm water for 8 hours, adding 0.2% edible alkali. Drain until the moisture content is 48-53%. Pear shells are ready for use. Mix the three types of crushed shells evenly and set aside.
[0054] Mixing ingredients: After crushing the soybean cake, add it to the mixing equipment in sequence with the pre-treated mixed crushed shells, bran, corn flour, gypsum powder, sucrose, superphosphate, magnesium sulfate, urea, kudzu root powder, and distiller's grains. Mix at low speed for 25 minutes at 160 r / min. Add water in stages to adjust the moisture content of the mixture to 59-64%. Mix until the material is uniform and free of lumps.
[0055] Fermentation treatment: The mixed material is piled into a heap 80cm high and 100cm wide, covered with plastic film, and fermented in stages. The first stage fermentation temperature is 28-32℃, fermentation lasts for 3 days, and the heap is turned once a day. The second stage fermentation temperature is 24-28℃, fermentation lasts for 4 days, and the heap is turned once every 2 days. During the fermentation process, 0.08% of fermentation inoculant is added. The fermentation inoculant is a compound inoculant of Bacillus subtilis and yeast in a mass ratio of 1:1. An appropriate amount of water is added to maintain the moisture content. Fermentation continues until the heap has no odor and turns yellowish-brown, thus obtaining the fermented material.
[0056] Sterilization and inoculation: Pack the fermentation material into polypropylene plastic bags, filling each bag to 70% of its volume. Sterilize using atmospheric pressure at 100℃ for 12-14 hours. After sterilization, cool to below 28℃. Inoculate with *Flammulina velutipes* spawn in a sterile environment, using 4% of the fermentation material's mass. Strictly adhere to aseptic procedures during inoculation. The aseptic colony count should be ≤10 CFU / cm³. 2 The enoki mushroom strain used is a commercially available conventional enoki mushroom cultivation strain.
[0057] Mycelium growth and cultivation: Place the inoculated bags into the cultivation room, control the temperature of the cultivation room to be 24-26℃, the relative humidity of the air to be 67-71%, maintain a dark environment, and the light intensity to be ≤50 lux. Ventilate once a day for 20 minutes each time. Cultivate for 33-38 days until the mycelium has fully grown into the bags and is fully mature, and then transfer to the fruiting management.
[0058] Mushroom management: Transfer mature mushroom bags to the mushroom growing room, control the temperature in the growing room at 10-13℃ and the relative humidity at 85-90%, and install a 15W light bulb every 3-5m to induce continuous weak light for 4 hours a day with a light intensity of 50-100 lux. Ventilate once a day for 15 minutes each time. Harvest when the stipe reaches 14-19cm in length and the cap diameter reaches 0.7-1.1cm.
[0059] Comparative Example 1: The existing traditional sawdust culture medium formula is used, without the use of crushed shells. The raw material components by weight are: 20 parts sawdust, 5 parts wheat bran, 3 parts corn flour, 1 part gypsum powder, 1 part sucrose, 0.5 parts superphosphate, 0.1 parts magnesium sulfate, 0.2 parts urea, and appropriate amount of water. The preparation method is completely consistent with that of Example 1.
[0060] Comparative Example 2: Using the existing single-shell utilization scheme, only nut shells are used, without adding livestock and poultry shells or fruit and vegetable shells. The raw material components by weight are: 25 parts nut shells, 8 parts wheat bran, 4 parts corn flour, 1.2 parts gypsum powder, 1.2 parts sucrose, 0.6 parts superphosphate, 0.15 parts magnesium sulfate, 0.3 parts urea, and appropriate amount of water. The preparation method is the same as in Example 2.
[0061] Comparative Example 3: The existing crushed shell pretreatment scheme is adopted. The crushed shells are not soaked, deoiled, or deacidified before being used directly for culture medium preparation. The raw material composition is completely consistent with that of Example 3. The crushed shell pretreatment step is omitted in the preparation method, and the rest of the operation is the same as that of Example 3.
[0062] Comparative Example 4: The existing traditional single fermentation process is adopted without staged fermentation. The raw material composition is completely consistent with that of Example 3. The fermentation step in the preparation method is changed to piling the mixture into a pile, covering it with plastic film, fermenting at 25-30℃ for 9 days, turning the pile once a day. The remaining operations are the same as those in Example 3.
[0063] Comparative Example 5: The proportion of crushed shells exceeds the scope of protection of this invention. At the same time, referring to the existing high proportion of crushed shells, the raw material components by mass are: 45 parts of crushed nut shells, 20 parts of crushed livestock and poultry shells, 15 parts of crushed fruit and vegetable shells, 16 parts of wheat bran, 6 parts of corn flour, 2 parts of gypsum powder, 2 parts of sucrose, 1.0 part of superphosphate, 0.25 parts of magnesium sulfate, 0.5 parts of urea, 2 parts of kudzu root powder, 5 parts of distiller's grains, 5 parts of soybean cake, and appropriate amount of water. The preparation method is the same as in Example 4.
[0064] Comparative Example 6: Using the existing single nutrient system scheme, without adding compound nutrient regulators, the raw material composition is the same as in Example 4, except that kudzu root powder, distiller's grains, and soybean cake are removed, and the rest of the operation is the same as in Example 4.
[0065] Performance testing and results analysis: To verify the technical effects of the embodiments of the present invention and the differences between them and existing technologies and their combinations, the enoki mushroom culture media prepared in Examples 1-5 and Comparative Examples 1-6 were subjected to performance tests. The test indicators included mycelial growth rate, mycelial growth cycle, fruiting time, yield per bag, crude protein content of enoki mushrooms, dietary fiber content, and contamination rate of miscellaneous bacteria. All test samples were tested under the same culture and fruiting environment to ensure the accuracy and comparability of the test results. The test methods and results are as follows.
[0066] Test method: Mycelial growth rate: After inoculation, observe and measure the mycelial growth length in the bag every day, calculate the average daily growth rate in mm / d, and continue to measure until the mycelium has fully covered the bag.
[0067] Mycelium growth cycle: The time required from the completion of inoculation to the mycelium fully growing on the bag and reaching full maturity, measured in days.
[0068] Mushroom emergence time: The time required from the start of mushroom emergence management to the appearance of the first batch of mushroom buds, measured in days.
[0069] Yield per bag: Each bag of enoki mushrooms is harvested 3 times, and the total weight is calculated and the average value is taken. The unit is g / bag.
[0070] Crude protein content: determined by the Kjeldahl method, in accordance with GB5009.5-2016.
[0071] Dietary fiber content: determined by enzymatic gravimetric method, in accordance with GB5009.88-2014.
[0072] Contamination rate: The percentage of contaminated bags out of the total number of bags, expressed as a percentage. The contamination criterion is the presence of non-Flammulina velutipes mycelium in the bags.
[0073] The test results are shown in Table 1 below:
[0074] Test Result Analysis: Compared with the existing traditional sawdust culture medium (Comparative Example 1), the mycelial growth rate of Examples 1 to 5 of the present invention is increased by 15.6% to 51.1%, the mycelial growth cycle is shortened by 5.3% to 26.3%, the yield per bag is increased by 12.5% to 59.4%, the crude protein content is increased by 8.4% to 30.8%, and the contamination rate of miscellaneous bacteria is reduced by 21.9% to 65.6%. This effectively solves the problems of scarce sawdust resources, high costs, and poor yield and quality in the existing technology, realizing the resource utilization of sawdust and having significant economic and environmental benefits. Compared with the existing single sawdust utilization scheme (Comparative Example 2), Examples 2 to 5 of the present invention, through the combination of composite sawdust, increase the mycelial growth rate by 13.7% to 33.3%, increase the yield per bag by 13.5% to 37.8%, and reduce the contamination rate of miscellaneous bacteria by 35.7% to 64.3%. This fully demonstrates the creativity of the present invention in the selection of composite sawdust and effectively solves the defects of the existing single sawdust, which has limited nutrition and poor performance. Compared to Comparative Example 3, which uses an existing crushed shell pretreatment scheme, Examples 3 to 5 of this invention, through differentiated pretreatment processes, achieve a 41.9% to 58.1% increase in mycelial growth rate, a 7.5% to 37.5% reduction in mycelial incubation cycle, and a 44.4% to 73.3% reduction in contamination rate. This fully demonstrates the inventiveness of the pretreatment process of this invention and effectively solves the shortcomings of existing crushed shell pretreatment methods, such as simplicity and high contamination rate. Compared to Comparative Example 4, which uses a traditional single fermentation scheme, Examples 3 to 5 of this invention, through a staged fermentation process, achieve a 35.4% to 39.6% increase in mycelial growth rate, a 7.7% to 28.2% reduction in mycelial incubation cycle, and a 45.5% to 54.5% increase in yield per bag. This fully demonstrates the inventiveness of the fermentation process of this invention and effectively solves the shortcomings of existing fermentation processes, such as long fermentation cycles and insufficient composting. Compared with the existing high-ratio crushed shell formulation (Comparative Example 5), the mycelial growth rate of Examples 4 to 5 of this invention increased by 25.6% to 53.8%, the mycelial growth cycle was shortened by 14.3% to 33.3%, the yield per bag increased by 24.1% to 110.3%, and the contamination rate of miscellaneous bacteria decreased by 38.5% to 80.8%. This fully demonstrates the creativity of the crushed shell ratio range design of this invention and effectively solves the defects of unreasonable crushed shell ratios and performance degradation in existing formulations. Compared with the existing single-nutrient system formulation (Comparative Example 6), Examples 4 to 5 of this invention, through the construction of a composite nutrient system, increased the mycelial growth rate by 16.0% to 36.0%, the yield per bag increased by 2.9% to 45.7%, and the crude protein content increased by 3.8% to 25.2%. This fully demonstrates the creativity of the nutrient system design of this invention and effectively solves the defects of single-nutrient systems and poor quality of enoki mushrooms in existing formulations.Furthermore, there is a clear progressive correlation between Examples 1 to 5. Example 2 increases the types of crushed shells and nutrient components based on the technical solution of Example 1, resulting in a significant performance improvement. Example 3 optimizes the fermentation process and adds auxiliary nutrients based on the technical solution of Example 2, further improving the performance. Example 4 expands the upper limit of crushed shell ratio and optimizes the pretreatment and sterilization process, achieving optimal performance. Example 5 expands the lower limit of crushed shell ratio, forming a complete ratio coverage, and its performance is still superior to the existing technology and each comparative example.
[0075] The preferred embodiments of the present invention disclosed above are merely illustrative of the invention. These preferred embodiments do not exhaustively describe all details, nor do they limit the invention to the specific implementations described. Clearly, many modifications and variations can be made based on the content of this specification. This specification selects and specifically describes these embodiments to better explain the principles and practical applications of the invention, thereby enabling those skilled in the art to better understand and utilize the invention.
Claims
1. A culture medium for enoki mushrooms using crushed shells as raw material, characterized in that, The raw materials, by weight, include: 5 to 30 parts crushed nut shells, 3 to 15 parts crushed livestock and poultry shells, 2 to 10 parts crushed fruit and vegetable shells, 4 to 16 parts wheat bran, 2 to 6 parts corn flour, 0.8 to 2 parts gypsum powder, 0.8 to 2 parts sucrose, 0.4 to 1.0 parts superphosphate, 0.08 to 0.25 parts magnesium sulfate, 0.1 to 0.5 parts urea, 0.5 to 2 parts kudzu root powder, 2 to 5 parts distiller's grains, 3 to 5 parts soybean cake, and an appropriate amount of water.
2. The enoki mushroom culture medium according to claim 1, characterized in that, The nut shells are one or more of walnut shells, peanut shells, and chestnut shells; the poultry shells are one or more of chicken shells, duck shells, and goose shells; and the fruit and vegetable shells are one or more of citrus shells, apple shells, and pear shells.
3. The enoki mushroom culture medium according to claim 1, characterized in that, The particle size of the crushed nut shells, livestock and poultry shells, and fruit and vegetable shells is 0.5 to 10 mm.
4. A method for preparing the *Flammulina velutipes* culture medium using crushed shells as raw material as described in claim 1, characterized in that, Includes the following steps: (1) Shell pretreatment: crush the shells of nuts, livestock and poultry, and fruits and vegetables to a particle size of 0.5 to 10 mm. Different types of shells are subjected to differentiated warm soaking or additive treatment. After draining to a moisture content of 48 to 57%, they are mixed evenly and set aside. (2) Mixing ingredients: Crush soybean cake, and then add the pre-treated mixed crushed shells, wheat bran, corn flour, gypsum powder, sucrose, superphosphate, magnesium sulfate, urea, kudzu root powder, distiller's grains, and crushed soybean cake to the mixing equipment in sequence. During the mixing process, add water to adjust the moisture content of the mixture to 59 to 67%, and stir until the material is uniform and free of lumps. (3) Fermentation treatment: The mixture is piled into a pile with a height of 80 to 90 cm and a width of 100 to 110 cm, covered with plastic film for staged fermentation. During the fermentation process, the pile is turned frequently and water is added to maintain the moisture content. Fermentation agents can be added. Fermentation continues until the pile has no odor, and fermented material is obtained. (4) Sterilization and inoculation: The fermentation material is put into a polypropylene plastic bag, the filling amount is 70% of the bag volume, and after sterilization, it is cooled to below 30°C. Inoculate the enoki mushroom spawn in a sterile environment with a colony count ≤10cfu / cm². (5) Mycelium growth and cultivation: Place the inoculated bags in the cultivation room, control the temperature, relative humidity, light intensity and ventilation conditions of the cultivation room, and cultivate until the mycelium has fully grown into the bags and is fully mature; (6) Mushroom management: Transfer mature mushroom bags to the mushroom growing room, control the temperature, relative humidity, light intensity and ventilation conditions of the mushroom growing room, and harvest when the enoki mushrooms grow to a stipe length of 14 to 23 cm and a cap diameter of 0.7 to 1.5 cm.
5. The preparation method according to claim 4, characterized in that, In step (1), nut shells are soaked in 50°C warm water for 12 to 36 hours, with the water changed twice, and drained until the moisture content is 48 to 57%; poultry shells are soaked in 60°C warm water for 8 to 12 hours, with 0.2 to 0.5% edible alkali added, and drained until the moisture content is 48 to 57%; fruit and vegetable shells are used directly or soaked in 0.2% citric acid for 6 hours and then drained until the moisture content is 48 to 57%.
6. The preparation method according to claim 4, characterized in that, In step (2), the mixing is either a single low-speed mixing or a combination of high-speed mixing and low-speed mixing. The mixing speed is 150 to 400 r / min, the total mixing time is 15 to 30 minutes, and the moisture content of the mixture is adjusted to 59 to 67%. When high-speed mixing and low-speed mixing are combined, high-speed mixing is performed for 10 to 15 minutes first, followed by low-speed mixing for 15 minutes.
7. The preparation method according to claim 4, characterized in that, In step (3), the height of the material pile is 80 to 90 cm and the width is 100 to 110 cm; the temperature of the first stage of the phased fermentation is 28 to 36℃, the fermentation lasts for 3 to 4 days, and the pile is turned over 1 to 2 times a day; the temperature of the second stage is 24 to 30℃, the fermentation lasts for 4 to 6 days, and the pile is turned over once every 2 days; the fermentation agent is a compound agent composed of Bacillus subtilis and yeast in a mass ratio of 1:1, and its addition amount is 0.08 to 0.2% of the mass of the mixture.
8. The preparation method according to claim 4, characterized in that, In step (4), the sterilization process is high pressure sterilization, normal pressure sterilization, or a combination of high pressure and normal pressure sterilization; the temperature of high pressure sterilization is 121℃ and the sterilization time is 1 to 2.5 hours, the temperature of normal pressure sterilization is 100℃ and the sterilization time is 12 to 16 hours; the inoculation amount of enoki mushroom spawn is 4 to 8% of the mass of the fermentation material.
9. The preparation method according to claim 4, characterized in that, In step (5), the temperature of the cultivation room is 21 to 26℃, the relative humidity is 65 to 72%, the light intensity is ≤50 lux, and the room is ventilated 1 to 2 times a day for 15 to 25 minutes each time. The cultivation time is 25 to 38 days. In step (6), the temperature of the fruiting room is 7 to 13℃, the relative humidity is 85 to 94%, the light intensity is 50 to 100 lux, and the room is ventilated 1 to 2 times a day for 10 to 25 minutes each time. The harvesting specifications for enoki mushrooms are a stem length of 14 to 23 cm and a cap diameter of 0.7 to 1.5 cm.