Methods for cultivating edible fungi strains using bark-covered wooden stalks

Abstract

This invention provides a method for cultivating edible fungi spawn using bark-covered wooden stems, belonging to the field of edible fungi production technology. The method includes: providing pretreated bark-covered wooden stems; providing a sawdust mixture; providing multiple polypropylene bags or spawn bottles to allow the bark-covered wooden stems and sawdust mixture to come into contact; dispensing and sterilizing the materials; inoculating the multiple polypropylene bags or spawn bottles with edible fungi mother spawn; and incubating at a constant temperature to obtain edible fungi spawn; providing cultivation bags containing sawdust mixture; inoculating the edible fungi spawn with the spawn; and incubating at a constant temperature to obtain edible fungi spawn. The bark-covered wooden stems are selected from the trunks or branches of trees with intact bark and a tight bond between the phloem and xylem. This method results in a short production and cultivation cycle for edible fungi spawn, uniform and synchronized spawn age, high inoculation success rate, and protection of the mycelium from damage during dispensing, reducing spawn production costs and improving spawn quality and yield.

Description

Technical Field

[0001] This invention relates to the field of edible fungi production technology, and in particular to a method for cultivating edible fungi strains using bark-covered wooden stalks. Background Technology

[0002] Edible fungi, such as wood ear mushrooms, lion's mane mushrooms, and button mushrooms, are a type of health food that is high in protein and low in fat. Edible fungi spores refer to strains isolated and screened from the large quantities of edible fungi in nature, then improved and stored for use in production. The traditional process of edible fungi production is from mother spore → primary spore → cultivation spore → cultivation; this method has become standardized. Therefore, edible fungi spores are divided into three types: mother spore, primary spore, and cultivation spore, also called primary, secondary, and tertiary spores, respectively. Mother spores are propagated in test tube culture media and are used for propagating primary spores and preserving seeds. Primary spores are propagated from mother spore mycelium on solid culture media; this process enhances the mycelium's adaptability to the culture environment and expands the propagation of edible fungi. Cultivation spores are expanded from primary spores and mainly provide sufficient spores for edible fungi production and cultivation.

[0003] Currently, the main types of spawn used in production and cultivation are sawdust spawn, branch spawn, and liquid spawn. Sawdust spawn is stable and resistant to contamination, but its production is labor-intensive, inefficient, and expensive. It is also susceptible to contamination by other microorganisms, has a long production cycle, and yields are unstable. Liquid spawn improves efficiency and reduces production time, making it suitable for factory production. However, it requires high standards for production and operation, involves significant investment, requires sterility throughout the process, and is difficult to store and transport. Branch spawn is easy to inoculate, has a consistent mycelial age, and can shorten the mycelial growth cycle, but requires pretreatment of the branches and a more cumbersome bagging process. In current edible mushroom production, sawdust is commonly used to cultivate spawn. However, the spawn needs to be used in blocks when inoculating, otherwise the mycelium is prone to breakage, resulting in slow germination and directly affecting the quality of the spawn. Furthermore, since the spawn is often placed in the pre-drilled holes of the cultivation bottle (bag) and cannot enter the bottom of the cultivation medium, the mycelium growth rate is also very slow. This often results in the upper mycelium being old while the lower mycelium is just beginning to grow, meaning there is a large difference in the age of the upper and lower mycelium, which in turn affects the production and cultivation arrangements and yield of edible mushrooms.

[0004] It is evident that the cultivation of edible fungi spores is a crucial factor in the production and development of edible fungi. Therefore, finding methods for cultivating edible fungi spores that are more conducive to spore germination, protect the mycelium from damage during propagation, and reduce spore production costs is an urgent problem that the edible fungi production industry needs to solve. Summary of the Invention

[0005] This invention provides a method for cultivating edible fungi spawn using bark-covered wooden stalks, which results in a short production and cultivation cycle, uniform and synchronized spawn ages, high inoculation success rate, protection of the mycelium from damage during division, reduced spawn production costs, and improved spawn quality and yield.

[0006] This invention provides a method for cultivating edible fungi spawn using bark-covered wooden stems, comprising: providing pretreated bark-covered wooden stems; providing a sawdust mixture; providing multiple polypropylene bags or spawn bottles to bring the bark-covered wooden stems and sawdust mixture into contact, dispensing and sterilizing the materials, inoculating the multiple polypropylene bags or spawn bottles with edible fungi mother spawn, and incubating at a constant temperature to obtain edible fungi spawn; providing cultivation bags containing sawdust mixture, inoculating with edible fungi spawn, and incubating at a constant temperature to obtain edible fungi spawn; the bark-covered wooden stems are selected from the trunks or branches of trees with intact bark and a tight bond between the phloem and xylem.

[0007] Through the above technical solution, the mycelium grows and reproduces both inside and outside the bark of the bark-covered wooden trunk. The amount of mycelium carried during inoculation is large, and after inoculation, the mycelium germinates at multiple points and spreads from the inside out, which shortens the production and cultivation cycle of edible fungi spawn, improves the utilization rate of mother spawn and original spawn, and the cultivated spawn has uniform age, stronger synchronization, better quality, enhanced anti-aging ability, improved production efficiency, reduced production costs, and can be used for large-scale production of edible fungi spawn.

[0008] The further steps involve the pretreatment of bark-covered wooden stems as follows: Cut tree trunks or branches into stems 0.5-0.8 cm in diameter and 8-15 cm in length, soaking them in a nutrient solution for 24-36 hours. Then, remove the stems and mix them thoroughly with fine bran. The soaking time should be until the stems are no longer white inside. Soaking the bark-covered stems in the nutrient solution allows them to fully absorb water and nutrients, resulting in dense, white mycelium and vigorous growth, achieving the standards for high-quality fungal strains.

[0009] Further configured, the weight ratio of bark-covered wooden stems to fine wheat bran is 100:(10-20); the fine wheat bran particle size is such that the residue on an 80-mesh sieve is no more than 50%. Preferably, the weight ratio of bark-covered wooden stems to fine wheat bran is 100:15. After soaking, the bark-covered wooden stems retain moisture and humidity on their surface. When mixed with fine wheat bran, the fine wheat bran adheres evenly to the surface of the bark-covered wooden stems, ensuring the stems are fully coated with the mixture. This facilitates nutrient absorption and growth of the mycelium, accelerating mycelial growth.

[0010] Further configured, the components and their weight ratio of the nutrient solution are: water:potassium dihydrogen phosphate:sugar = 100:(0.2-0.5):(0.1-0.3). Preferably, the nutrient solution consists of 100 catties of water, 0.2 catties of potassium dihydrogen phosphate, and 0.1 catties of sugar.

[0011] Further, the preparation process of the wood chip composite is as follows: 80-85 catties of wood chips and 15-20 catties of wheat bran are mixed to form a mixture. Then, 1-1.5% of white sugar, 0.8-1% of gypsum, and 1-1.5% of potassium dihydrogen phosphate are added to the mixture and stirred until well mixed. Water is then added to adjust the moisture content to 60-70%. Preferably, the wood chip composite contains 80 catties of wood chips and 20 catties of wheat bran, and 1% of white sugar, 1% of gypsum, and 1% of potassium dihydrogen phosphate are also added to the mixture.

[0012] Further, the packaging process is as follows: Place a 1-2cm layer of sawdust mixture at the bottom of the polypropylene bag or culture bottle, then neatly place the bark-covered wooden sticks into the polypropylene bag or culture bottle, fill the gaps between the bark-covered wooden sticks with sawdust mixture, and finally place another 1-2cm layer of sawdust mixture on top of the bark-covered wooden sticks, and seal with a film.

[0013] The sawdust mixture is filled into the top, bottom, and gaps of the bark-covered wooden stalks. This sawdust mixture acts as a bridging medium, serving as a growth medium for mycelium, allowing newly formed mycelium to quickly grow into the bark-covered stalks and accelerating mycelial growth. Simultaneously, the sawdust mixture provides nutrients, which is beneficial for mycelial colonization. Furthermore, filling the gaps with sawdust mixture makes it easier to separate the bark-covered stalks when they are removed, thus preserving the original mycelium as much as possible.

[0014] Further settings include: sterilization at normal pressure or autoclave; normal pressure sterilization is maintained at 100℃ for 8-10 hours; autoclave sterilization is maintained at 120-125℃ for 1.5-2.5 hours.

[0015] The inoculation process for edible fungi spawn is further configured as follows: In a sterile environment, a wooden stalk with bark containing the edible fungi spawn is inserted directly into the hole in the center of the sawdust mixture at one end of the cultivation bag, ensuring that the stalk does not protrude above the substrate surface, and one stalk is inserted into each cultivation bag. The constant temperature incubation conditions are further configured as follows: dark environment, temperature 20-28℃, time 15-25 days.

[0016] By using bark-covered wooden sticks with edible fungi spawn for inoculation and cultivation, and with the sawdust mixture in the cultivation bag having holes in the middle, the bark-covered wooden sticks are easily inserted into the cultivation bag, ensuring sufficient oxygen supply from top to bottom, which accelerates the mycelial growth rate. This also reduces the damage to mycelium and low mycelial growth rate caused by forcibly inserting bark-covered wooden sticks into the cultivation bag. The suitable temperature cultivation allows the mycelium to fully consume the substrate and grow at multiple points, resulting in snow-white, robust mycelium of the cultivated spawn with strong resistance to contamination by other microorganisms. This also shortens the spawn production cycle and reduces costs.

[0017] The present invention also provides an edible fungus strain, which is prepared by the above-described method of cultivating edible fungus strains using bark-covered wooden stalks.

[0018] The method for cultivating edible fungi spawn using bark-covered wooden stalks provided by this invention achieves the following beneficial effects compared with existing technologies:

[0019] 1) Using bark-covered wooden stalks to cultivate edible fungi spawn provides good permeability, rapid mycelial growth, and shortens the cultivation time by 15-30 days compared to conventional sawdust cultivation. The mother culture has a high survival rate, grows rapidly, and has a low contamination rate, effectively shortening the cultivation time of the spawn. This results in mycelial growth that is vigorous, uniform, and dense, and also significantly enhances its anti-aging ability after inoculation.

[0020] 2) The cultivation spawn is inoculated and cultivated using bark-covered wooden stalks with original edible fungi spawn, combined with hole-punching inoculation. The mycelium grows and reproduces both inside and outside the bark of the bark-covered wooden stalks, resulting in a larger amount of mycelium carried during inoculation, thus increasing the inoculation volume. After inoculation, the mycelium germinates at multiple points and spreads from the inside out, and the time to full coverage of the bag can be 5-10 days earlier than conventional branch spawn, shortening the production and cultivation cycle of edible fungi spawn, improving production efficiency, and the cultivated spawn has a uniform age, stronger synchronization, and better quality, which is beneficial for the next stage of cultivation.

[0021] 3) This invention is applicable to the production and cultivation of most edible fungi such as oyster mushrooms, Ganoderma lucidum, shiitake mushrooms, wood ear fungus, tea tree mushrooms, and king oyster mushrooms. The process is simple, the inoculation success rate is high, the mycelium growth rate is fast, the utilization rate of mother culture and original culture is high, the production cost is low, and it can be used for large-scale production of edible fungi spawn. Moreover, the size of the bark-covered wooden poles can be changed according to production requirements, making production flexible and not wasteful of materials. The scope of application is greatly expanded and extended, which is conducive to mass production. Attached Figure Description

[0022] To more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the drawings used in the description of the embodiments or the prior art will be briefly introduced below. Obviously, the drawings described below are some embodiments of the present invention. For those skilled in the art, other drawings can be obtained based on these drawings without creative effort.

[0023] Figure 1 A graph showing the changing trends of polyphenol oxidase activity in king oyster mushrooms treated with different methods at different time periods. Detailed Implementation

[0024] To make the objectives, technical solutions, and advantages of the embodiments of the present invention clearer, 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 also within the scope of protection of the present invention.

[0025] Unless otherwise specified, all test materials and reagents used in the following examples are commercially available. Where specific techniques or conditions are not specified in the examples, they can be performed according to the techniques or conditions described in the literature or the product instructions.

[0026] In specific implementation, a method for cultivating edible fungi spawn using bark-covered wooden stalks includes the following steps:

[0027] 1) Pretreatment of bark-inclusive wood poles: Select tree trunks or branches of trees with intact bark and tight bonding between phloem and xylem, and then make bark-inclusive wood poles with a diameter of 0.5-0.8cm and a length of 8-15cm. Soak the bark-inclusive wood poles in nutrient solution for 24-36 hours, then take them out and mix them evenly with fine bran for later use.

[0028] The trunks or branches mentioned above are taken from fresh or dried trees. The trees should be 1-2 years old and in their dormant period. Trees from the families Betulaceae, Betulaceae, Ligustrum, and Ficus are preferred, but other broad-leaved species are also acceptable. Hardwoods are better than softwoods.

[0029] It should be noted that the bark-covered wooden poles in this invention refer to wooden poles that are made with bark in each pole during the pole preparation process. Wooden poles made from the xylem of the tree trunk that does not have bark are not used.

[0030] 2) Dispensing and sterilization: Dispense the bark-covered wood stalks and sawdust mixture into polypropylene bags or inoculum bottles, seal them, and sterilize them. After the polypropylene bags or inoculum bottles have cooled naturally to 20-25℃, they are ready for inoculation.

[0031] The number of bark-covered wooden stalks placed in the polypropylene bag or culture bottle should be sufficient to fill the bag or bottle. In a specific example, each bag / bottle contains 50-300 bark-covered wooden stalks.

[0032] It should be noted that the length of the wooden stick with bark varies with the length of the polypropylene bag or the culture bottle, and the length of the wooden stick with bark should be 2-3 cm shorter than the length of the polypropylene bag or the culture bottle. In a specific example of this invention, the size of the culture bottle is 750-1000 mL; the size of the polypropylene bag is 14 cm wide × 33 cm long and 0.5 mm thick.

[0033] 3) Primary Culture: Under aseptic conditions, inoculate the edible mushroom mother culture into polypropylene bags or spawn bottles, and incubate in the dark at a constant temperature of 20-28℃ for 15-25 days. Once the mycelium has fully grown, the primary culture of edible mushrooms is obtained. Using bark-covered wooden stalks for primary culture results in a high inoculation survival rate, rapid mycelial growth, and low contamination rate. After-ripening allows the mycelium to fully penetrate the bark-covered wooden stalks, and the bark-covered wooden stalks provide more mycelial growth points during the cultivation of spawn, which is beneficial for accelerating the cultivation of spawn.

[0034] It should be noted that when inoculating the mother culture, methods include, but are not limited to, inoculation at one or both ends, or ventral inoculation (ventral inoculation is not recommended because it is prone to contamination by multiple openings).

[0035] 4) Spawn Cultivation: Fill cultivation bags with sawdust-based mixed substrate, sterilize and cool them down. Insert one bark-covered wooden stalk containing the original edible fungus spawn into each cultivation bag. Then, place the cultivation bags in the dark and incubate at a constant temperature of 20-28℃ for 15-25 days to obtain the edible fungus spawn. The sawdust-based mixed substrate mentioned above is the same substrate used in the original spawn cultivation.

[0036] It should be noted that the cultivation bags are filled with a mixture of sawdust and substrate using a bagging machine equipped with an automatic perforation device. During the inoculation of the edible fungi spawn, in a sterile room / box, each bark-covered wooden stalk is pulled out one by one and inserted directly into the hole in the center of the sawdust mixture at one end of the cultivation bag, ensuring that the bark-covered stalks do not protrude above the substrate surface. Furthermore, while this invention uses bagged substrate for spawn production, it is not limited to the use of cultivation bags and can also be applied to the production of spawn using spawn bottles.

[0037] The inventors unexpectedly discovered that in the existing technology, when cultivating branch spawn, the branches with bark are not specifically selected as the inoculation material; in fact, even barkless chopsticks are used directly for inoculation. Therefore, when the original spawn is inoculated into the cultivation bag, the mycelium attached to the branch surface rubs against the bag material, easily damaging the mycelium on the branch surface. This causes some mycelium to break, which intensifies the respiration of the spawn, easily generating a large amount of heat and moisture, and thus easily causing mycelial burn and death. This results in inconsistent mycelial growth time in the cultivation bag, and even under the same cultivation time, there will be local areas without mycelium.

[0038] Based on this, the present invention specifically uses bark-covered wooden sticks when cultivating edible fungi spawn. During the growth of the original spawn, the mycelium can penetrate deep between the xylem and phloem. When inoculated into the cultivation bag, the bark / phloem protects the mycelium, reducing mycelial damage. In the cultivation bag, the mycelium can quickly adapt to the environment and develop, spreading from under the bark and from the inside out. Therefore, the inoculation success rate is higher, the mycelium age after inoculation is shorter and more uniform, the spawn grows faster and reproduces better, and the cultivation time is shortened by 5-10 days compared to the cultivation time of general branch spawn.

[0039] Based on this, the present invention also provides the use of bark-infused wooden poles in the cultivation of edible fungi spawn. The bark-infused wooden poles are used for inoculating and cultivating original or cultivated edible fungi spawn, and the bark-infused wooden poles are selected from the trunks or branches of trees with intact bark and a tight bond between the phloem and xylem.

[0040] The bark-covered wooden sticks of the present invention can not only be used to cultivate primary spawn as in the present invention, but also to cultivate cultivars using a similar method to the primary spawn cultivation method of the present invention. Then, the bark-covered wooden sticks with cultivar mycelium are inserted into mushroom bags, mushroom packs or mushroom sticks for the production of fruiting bodies of edible fungi.

[0041] As a further improvement to the aforementioned implementation, the preparation process of the sawdust mixture for cultivating cultivars is as follows: 80-85 catties of sawdust and 15-20 catties of wheat bran are mixed to form a mixture. Then, 1-1.5% white sugar, 0.8-1% gypsum, 1-1.5% potassium dihydrogen phosphate, 0.1-0.5% linoleic acid, and 0.5-1% castor oil are added to the mixture and stirred until well combined. Water is then added to adjust the moisture content to 60-70%. The linoleic acid and castor oil in the improved sawdust mixture work synergistically to effectively activate polyphenol oxidase during mycelial growth, resulting in an overall increase in polyphenol oxidase activity. The polyphenol oxidase secreted into the sawdust mixture decomposes lignin, thus accelerating the degradation rate of lignin and other macromolecules. The decomposition products are nutrients for mycelial growth. Sufficient nutrients enhance the mycelial growth rate, resulting in better mycelial growth and greater mycelial biomass, playing a crucial role in promoting rapid mycelial growth and shortening the cultivation time.

[0042] The present invention will be further described in detail below with reference to embodiments. However, it should be understood that the embodiments are for illustrative purposes only and are not intended to limit the scope of the present invention.

[0043] Example 1:

[0044] A method for cultivating edible fungi spawn using bark-covered wooden stalks includes the following steps:

[0045] 1) Select tree trunks or branches from trees with intact bark and a tight bond between the phloem and xylem. Then, prepare bark-inclusive wooden poles 0.5-0.8 cm in diameter and 10-15 cm in length. Soak these poles in a nutrient solution for 24 hours, then remove them and mix them thoroughly with fine wheat bran at a weight ratio of 100:20. The fine wheat bran should have a particle size where the residue on an 80-mesh sieve is no more than 50%.

[0046] The components and their weight ratio of the above nutrient solution are: water:potassium dihydrogen phosphate:sugar = 100:0.3:0.2. The above tree trunks or branches are taken from fresh trees or dried trees, which are 1-2 years old and have entered dormancy.

[0047] 2) Distribute the bark-covered wooden stalks and sawdust mixture into polypropylene bags or inoculum bottles. When distributing, place a 1-2cm layer of sawdust mixture at the bottom of the polypropylene bag or inoculum bottle, then neatly place the bark-covered wooden stalks into the polypropylene bag or inoculum bottle, and fill the gaps between the bark-covered wooden stalks with sawdust mixture. Finally, place another 1-2cm layer of sawdust mixture on top of the bark-covered wooden stalks, cover and seal with film, and autoclave at 121℃ for 2 hours. After the polypropylene bag or inoculum bottle has cooled naturally to 20-25℃, it is ready for inoculation.

[0048] The preparation process of wood chip composite is as follows: Mix 83 catties of wood chips and 17 catties of wheat bran to form a mixture. Then add 1.3% white sugar, 0.8% gypsum and 1.1% potassium dihydrogen phosphate to the mixture and mix well. Then add water to adjust the moisture content to 60-70%.

[0049] 3) Under aseptic conditions, inoculate the edible fungus mother culture into a polypropylene bag or spawn bottle, place it in the dark, and incubate at a constant temperature of 20-28℃ for 15-25 days. Once the mycelium has fully grown, the original edible fungus spawn is obtained.

[0050] 4) Using a bagging machine with an automatic punching device, fill the cultivation bags with the sawdust mixture prepared in step 2). After sterilization and cooling, in a sterile environment, take a bark-covered wooden stalk containing the edible fungus spawn and insert it straight into the hole in the middle of the sawdust mixture in the cultivation bag from the center of one end, ensuring that the bark-covered wooden stalk does not protrude from the surface of the spawn. Insert one bark-covered wooden stalk into each cultivation bag. Then place the cultivation bags in the dark and at a constant temperature of 20-28℃ for 15-25 days to obtain the edible fungus spawn.

[0051] Example 2:

[0052] A method for cultivating edible fungi spawn using bark-covered wooden stalks includes the following steps:

[0053] 1) The difference from step 1) in Example 1 is that the nutrient solution consists of 100 catties of water, 0.2 catties of potassium dihydrogen phosphate, and 0.1 catties of white sugar. The bark-covered wooden poles and fine bran are mixed evenly at a weight ratio of 100:15.

[0054] 2) The difference from step 2) in Example 1 is that the mixture of sawdust composite material contains 80 catties of sawdust and 20 catties of bran, and 1% of white sugar, 1% of gypsum and 1% of potassium dihydrogen phosphate are added to the mixture.

[0055] 3) Same as step 3) in Example 1.

[0056] 4) Same as step 4) in Example 1.

[0057] Example 3:

[0058] A method for cultivating edible fungi spawn using bark-covered wooden stalks includes the following steps:

[0059] 1) Same as step 1) in Example 2.

[0060] 2) Same as step 2) in Example 2.

[0061] 3) Same as step 3) in Example 2.

[0062] 4) The difference from step 4) in Example 2 is that the preparation process of the sawdust mixture for cultivating cultivars is as follows: 80 catties of sawdust and 20 catties of wheat bran are mixed to form a mixture. Then, 1% white sugar, 1% gypsum, 1% potassium dihydrogen phosphate, 0.3% linoleic acid and 0.5% castor oil are added to the mixture and mixed well. Water is then added to adjust the moisture content to 60-70%.

[0063] Comparative Example 1:

[0064] A method for cultivating edible fungi strains differs from Example 2 only in that clean disposable chopsticks are used for strain cultivation, instead of using wooden stalks with peels.

[0065] Comparative Example 2:

[0066] A method for cultivating edible fungi strains differs from Example 2 only in that: bark-free wooden poles made from the xylem trunks or branches are used for strain cultivation, instead of bark-covered wooden poles.

[0067] Comparative Example 3:

[0068] A method for cultivating edible fungi strains differs from Example 2 only in that: only sawdust composite material is used for strain cultivation, without using bark-covered wooden stalks, which is the sawdust cultivation method in the prior art.

[0069] Comparative Example 4:

[0070] A method for cultivating edible fungi spawn using bark-covered wooden stalks differs from Example 3 only in that: in step 4), the preparation process of the sawdust mixture used for cultivating the spawn is as follows: 80 catties of sawdust and 20 catties of wheat bran are mixed to form a mixture, then 1% white sugar, 1% gypsum, 1% potassium dihydrogen phosphate, and 0.8% linoleic acid are added to the mixture and mixed well, and then water is added to adjust the moisture content to 60-70%.

[0071] Comparative Example 5:

[0072] A method for cultivating edible fungi spawn using bark-covered wooden stalks differs from Example 3 only in that: in step 4), the preparation process of the sawdust mixture used for cultivating the spawn is as follows: 80 catties of sawdust and 20 catties of wheat bran are mixed to form a mixture, then 1% white sugar, 1% gypsum, 1% potassium dihydrogen phosphate, and 0.8% castor oil are added to the mixture and stirred well, and then water is added to adjust the moisture content to 60-70%.

[0073] Comparative Example 6:

[0074] A method for cultivating edible fungi spawn using bark-covered wooden stalks differs from Example 3 only in that: in step 4), the preparation process of the sawdust mixture used for cultivating the spawn is as follows: 80 catties of sawdust and 20 catties of wheat bran are mixed to form a mixture, then 1% white sugar, 1% gypsum, 1% potassium dihydrogen phosphate, 0.3% linoleic acid and 1.2% castor oil are added to the mixture and stirred well, and then water is added to adjust the moisture content to 60-70%.

[0075] Comparative Example 7:

[0076] A method for cultivating edible fungi spawn using bark-covered wooden stalks differs from Example 3 only in that: in step 4), the preparation process of the sawdust mixture used for cultivating the spawn is as follows: 80 catties of sawdust and 20 catties of wheat bran are mixed to form a mixture, then 1% white sugar, 1% gypsum, 1% potassium dihydrogen phosphate, 0.8% linoleic acid and 0.3% castor oil are added to the mixture and stirred well, and then water is added to adjust the moisture content to 60-70%.

[0077] Experimental Example 1: The Effect of Different Methods on the Growth of Microbial Strains

[0078] Experimental Methods: *Auricularia auricula-judae* and *Lentinula edodes* were cultured according to the methods described in Examples 1-3 and Comparative Examples 1-7, respectively. The mother culture for *Auricularia auricula-judae* was Qin Dan No. 1, and for *Lentinula edodes*, it was Shen Xiang No. 4, both purchased from the Institute of Fungi / Edible Fungi. The original spawn was cultured in 14cm × 33cm polypropylene bags with a uniform filling height, single-head inoculation, and 50 bags per group. The cultivated spawn was also produced in 50 bags per group, with 3 replicates per group. During the cultivation of the cultivated spawn, the growth rate and vigor of the mycelium were observed and recorded after it reached the surface of the cultivation bag. Mycelial growth was defined as the color, density, and thickness of the mycelium. Mycelial density was graded into 5 levels, with 0 being the worst and ++++ being the best. Multiple people were randomly selected to score the density based on sensory evaluation without prior knowledge of the mycelium, and the average value was calculated. Time to full coverage (days): The time from inoculation to the mycelium completely covering the cultivation bag was considered. Since the time to full coverage varied within the same batch, the average value was used. The results are shown in Table 1.

[0079] Table 1. Growth of Auricularia auricula-judae and Shiitake mushroom spawn under different methods

[0080]

[0081] Observations revealed that during the cultivation of the fungus and shiitake mushroom spawn in Examples 1-3, the mycelium grew vigorously and exhibited the best growth. After the mycelium filled the bag, the culture medium inside was no longer visible. The edges were neat and strong, and the mycelium consumed the medium quickly. When the bag was full, the wooden stem was cut open, and the mycelium had spread to the spongy tissue inside. The mycelium on the surface of the stem was white. Among them, the mycelium growth in Example 3 was the best.

[0082] Comparing Examples 1-3 and Comparative Examples 1-3, the difference in the time to full coverage of the bag is quite significant. The time for mycelium to fully cover the bag in the branch culture is significantly shorter than that in the sawdust culture. Moreover, the time for mycelium to fully cover the bag in the bark-infused wooden stalk culture of the present invention is the shortest. This is because the bark-infused wooden stalk can protect the original mycelium during inoculation, reducing mycelial damage. It can quickly adapt to the environment and develop in the cultivation bag, thus resulting in a higher inoculation success rate, faster mycelial growth and better propagation after inoculation, stronger synchronization, and better quality.

[0083] Comparing Examples 2-3 and Comparative Examples 4-7, it was found that the method in Example 3 had the shortest time for mycelium to fill the bag, and the synergistic effect of linoleic acid and castor oil in the sawdust composite material of Example 3 was significant. The differences between Comparative Examples 4 and 5 and Example 2 were not significant. Comparative Examples 6 and 7 showed that linoleic acid and castor oil in a specific ratio can promote rapid mycelial growth and shorten the cultivation time.

[0084] Experimental Example 2: Effects of different methods on enzyme activity and growth of Pleurotus ostreatus

[0085] Experimental Methods: *Pleurotus eryngii* were cultured according to the methods described in Examples 2-3 and Comparative Examples 4-7. The mother culture of *Pleurotus eryngii* was "Xingbao No. 6" provided by Guizhou Guiwang Biotechnology Co., Ltd. During the cultivation period, mycelial growth and time to full colony filling were observed and recorded according to the method described in Experiment 1. Simultaneously, the polyphenol oxidase activity of mycelia was measured at 5, 10, 15, and 20 days after inoculation. Polyphenol oxidase assay: A polyphenol oxidase kit from Shanghai Jizhi Biochemical Technology Co., Ltd. was used. Three biological replicates were performed for each treatment. Enzyme activity was defined as a change of 0.005 in absorbance at 410 nm per minute per mg of tissue per ml of reaction system. The calculation formula is as follows: Polyphenol oxidase activity (U / g mass) = △A ÷ 0.005 × Vreaction total ÷ (W × Vsample ÷ Vsample total) ÷ T = 120 × △A ÷ W, where, Vreaction total: total volume of the reaction system, 0.3 mL; Vsample: volume of sample added to the reaction system, 0.05 mL; Vsample total: volume of extraction solution added, 1 mL; W: sample mass, g; T: reaction time, 10 min. The results are shown in Table 2. Figure 1 As shown.

[0086] Table 2. Growth of King Oyster Mushrooms under Different Methods

[0087] Full bag time d density mycelial growth Example 2 22 ++++ White and robust Example 3 17 ++++ White and robust Comparative Example 4 21 ++++ White and robust Comparative Example 5 21 ++++ White and robust Comparative Example 6 24 +++ White and relatively thick Comparative Example 7 21 ++++ White and robust

[0088] The comparison revealed that the method in Example 3 had the shortest time to fully fill the bag with mycelium and the best growth; Comparative Examples 4, 5, and 7 were slightly better than Example 2, but the overall difference was not significant; Comparative Example 6 was worse than Example 2.

[0089] Figure 1 The graph shows the trend of polyphenol oxidase activity in *Pleurotus eryngii* treated by different methods at different stages. The results show that polyphenol oxidase activity initially increased and then decreased. Example 3 showed the highest overall enzyme activity and the fastest increase, reaching a peak of 110.54 U / g, followed by the slowest decrease, finally dropping to 99.58 U / g. Example 2 showed the highest enzyme activity, reaching 98.32 U / g, before decreasing to 84.36 U / g. Comparative Examples 4, 5, and 7 showed slightly higher enzyme activities than Example 2, but the overall differences were not significant. Comparative Example 6 showed even lower enzyme activity than Example 2, reaching a peak of only 93.72 U / g and then decreasing to 77.27 U / g. Mycelial development in Comparative Example 6 was inhibited compared to Example 2.

[0090] In summary, this demonstrates that linoleic acid and castor oil in the sawdust composite of Example 3 have a significant synergistic effect. Under specific ratios, linoleic acid and castor oil can effectively activate polyphenol oxidase during the mycelial growth stage, resulting in an overall increase in polyphenol oxidase activity. The polyphenol oxidase secreted into the sawdust composite decomposes lignin, thus accelerating the degradation rate of lignin and other macromolecules. The decomposition products are nutrients for mycelial growth. Sufficient nutrients enhance the mycelial growth rate, resulting in better mycelial growth, promoting rapid mycelial growth, and shortening the cultivation time.

[0091] It should be noted that, in this invention, unless otherwise specified, concentrations, ratios, etc. are all weight concentrations, weight ratios, etc., which are common writing habits of those skilled in the art, and therefore will not be described in detail in this invention.

[0092] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention, and not to limit them; although the present invention has been described in detail with reference to the foregoing embodiments, those skilled in the art should understand that modifications can still be made to the technical solutions described in the foregoing embodiments, or equivalent substitutions can be made to some or all of the technical features; and these modifications or substitutions do not cause the essence of the corresponding technical solutions to deviate from the scope of the technical solutions of the embodiments of the present invention.

Claims

1. A method for cultivating edible fungi spawn using bark-covered wooden stalks, characterized in that, include: Pre-treated, skinned wooden poles are available; Provide 1 unit of wood chip composite material; Multiple polypropylene bags or inoculum bottles are provided to allow the above-mentioned bark-covered wooden sticks and the sawdust composite material 1 to come into contact. After being packaged and sterilized, edible fungi mother culture is inoculated into the multiple polypropylene bags or inoculum bottles and cultured at a constant temperature to obtain edible fungi original culture. A cultivation bag containing sawdust composite material 2 is provided. The edible fungus spawn is inoculated and cultured at a constant temperature to obtain edible fungus spawn. The preparation process of the sawdust composite material 2 used for culturing the spawn is as follows: 80-85 catties of sawdust and 15-20 catties of wheat bran are mixed to form a mixture. Then, 1-1.5% of white sugar, 0.8-1% of gypsum, 1-1.5% of potassium dihydrogen phosphate, 0.1-0.5% of linoleic acid and 0.5-1% of castor oil are added to the mixture and stirred evenly. Water is then added to adjust the moisture content to 60-70%. The bark-covered wooden poles are selected from the trunks or branches of trees with intact bark and a tight bond between the phloem and xylem; The pretreatment process of the bark-inclusive wooden poles is as follows: the tree trunks or branches are made into bark-inclusive wooden poles with a diameter of 0.5-0.8cm and a length of 8-15cm. The bark-inclusive wooden poles are soaked in nutrient solution for 24-36 hours, then taken out and mixed evenly with fine bran for later use. The term "bark-covered wooden pole" refers to the requirement that each wooden pole obtained during the pole preparation process must have bark attached; wooden poles made from the xylem of the tree trunk without bark are not accepted. The inoculation process of the edible fungus spawn is as follows: In a sterile environment, take a wooden stick with the edible fungus spawn and insert it straight into the hole in the middle of the sawdust composite material 2 of the cultivation bag from the center of one end of the cultivation bag, ensuring that the wooden stick with the spawn does not protrude from the surface of the material, and insert one wooden stick with the spawn into each cultivation bag.

2. The method for cultivating edible fungi spawn using bark-covered wooden stalks according to claim 1, characterized in that, The weight ratio of the bark-covered wooden poles to the fine bran is 100:(10-20); the fine bran has a particle size of no more than 50% residue on an 80-mesh sieve.

3. The method for cultivating edible fungi spawn using bark-covered wooden stalks according to claim 1, characterized in that, The components and their weight ratio of the nutrient solution are: water:potassium dihydrogen phosphate:sugar = 100:(0.2-0.5):(0.1-0.3).

4. The method for cultivating edible fungi spawn using bark-covered wooden stalks according to claim 1, characterized in that, The preparation process of the wood chip composite material 1 is as follows: 80-85 catties of wood chips and 15-20 catties of wheat bran are mixed to form a mixture. Then, 1-1.5% of white sugar, 0.8-1% of gypsum and 1-1.5% of potassium dihydrogen phosphate are added to the mixture and stirred evenly. Water is then added to adjust the moisture content to 60-70%.

5. The method for cultivating edible fungi spawn using bark-covered wooden stalks according to claim 1, characterized in that, The packaging process is as follows: a 1-2cm layer of the sawdust composite material 1 is placed at the bottom of the polypropylene bag or the inoculum bottle, then the bark-covered wooden rods are neatly placed into the polypropylene bag or the inoculum bottle, and the gaps between the bark-covered wooden rods are filled with sawdust composite material 1. Finally, another 1-2cm layer of sawdust composite material 1 is placed on top of the bark-covered wooden rods, and the bag is sealed with a film.

6. The method for cultivating edible fungi spawn using bark-covered wooden stalks according to claim 1, characterized in that, The sterilization is performed under normal pressure or under high pressure; the normal pressure sterilization is performed at 100°C for 8-10 hours; the high pressure sterilization is performed at 120-125°C for 1.5-2.5 hours.

7. The method for cultivating edible fungi spawn using bark-covered wooden stalks according to claim 1, characterized in that, The constant temperature culture conditions are as follows: dark environment, temperature of 20-28℃, and time of 15-25 days.

8. An edible fungus strain, characterized in that, It is prepared by the method of cultivating edible fungi strains using bark-covered wooden stalks as described in any one of claims 1-7.

Images