Formula and method of liquid strain for improving yield of shiitake mushrooms

Abstract

The invention discloses a formula and method of a liquid strain for increasing the yield of shiitake mushrooms, and the method comprises the following steps: (1) inoculating the liquid strain into solid mushroom sticks by a pulse type high-pressure injection method under the injection pressure of 0.3-0.5 MPa, injecting each mushroom stick into 4-6 point positions, and injecting 3-5mL into each point position, and (2) culturing the inoculated mushroom sticks in a dark place under the conditions of 22-25 DEG C and humidity of 50-65%, the invention aims to solve the problems of low mycelium activity, long period and high pollution rate in the prior art. According to the technical scheme, the formula and the method of the liquid strain for improving the yield of the shiitake mushrooms are provided. Through the synergistic effect of the composite carbon and nitrogen source and growth promoting factors (nano silicon dioxide and vitamin B1), the hypha metabolic activity is improved by 40%-50%, the biomass accumulation rate is increased, the problem of slow growth caused by insufficient growth promoting factors in a traditional formula is solved, and the whole-process parameters (temperature, humidity and illumination) are digitally monitored, so that the production efficiency is improved. The repeatability and the stability of industrial production are realized.

Description

Technical Field

[0001] This invention relates to the field of liquid fermentation technology for edible fungi, and more specifically, to a formula and method for liquid spawn to increase the yield of shiitake mushrooms. Background Technology

[0002] Shiitake mushrooms, also known as fragrant mushrooms, winter mushrooms, or flower mushrooms, are a widely cultivated and consumed edible fungus. They belong to the kingdom Fungi, phylum Basidiomycota, class Agaricales, order Agaricales, family Tricholomataceae, genus Lentinus. Shiitake mushrooms are loved not only for their unique flavor but also for their nutritional and medicinal value, making them widely consumed globally.

[0003] A search revealed that Chinese patent document CN 112806215 A discloses a method for preparing liquid spawn for shiitake mushrooms and a corresponding production method for mushroom substrate, including the following specific steps: (1) preparation of secondary liquid spawn culture medium; (2) sterilization and inoculation of secondary liquid spawn; (3) cultivation of secondary liquid spawn; (4) preparation of tertiary solid high-quality mushroom substrate; (5) sterilization and inoculation of tertiary solid high-quality mushroom substrate; and (6) cultivation of tertiary solid high-quality mushroom substrate. This invention uses a novel spawn formula for the cultivation of secondary liquid spawn for shiitake mushrooms, which has advantages such as short cultivation time, high purity, and strong vitality. It also results in a high survival rate after inoculation, rapid mycelial growth, shortened cultivation time for tertiary solid high-quality mushroom substrate, reduced labor costs, and improved yield and quality of shiitake mushrooms. The existing technologies mentioned above have the following drawbacks in use: the carbon source in the disclosed formula is mainly corn flour and brown sugar, the nitrogen source depends on wheat bran, and there is a lack of growth-promoting factors, which means that there is still room for improvement in mycelial growth rate and metabolic activity. Traditional shaker culture requires 5 days (see step 3 in the comparative document), and it does not combine light regulation technology, so the mycelial biomass accumulation is slow. The existing sterilization and inoculation processes rely on manual operation, which is easy to introduce contamination (steps 2 and 4 in the comparative document), and there is a lack of real-time environmental monitoring methods.

[0004] Therefore, a new solution is needed to address this problem. Summary of the Invention

[0005] In view of the shortcomings of the existing technology, the purpose of this invention is to provide a formula and method for liquid spawn to increase the yield of shiitake mushrooms. Through formula optimization, photobiological regulation and process innovation, the problems of low mycelial activity, long cycle and high contamination rate in the existing technology are systematically solved.

[0006] The above-mentioned technical objective of the present invention is achieved through the following technical solution: a formula for a liquid spawn to increase the yield of shiitake mushrooms, characterized in that it comprises the following components by mass percentage: 8%-12% corn flour, 5%-8% brown sugar, 2%-4% glucose, 1%-2% wheat bran, 0.4%-0.6% potassium dihydrogen phosphate, 0.15%-0.25% magnesium sulfate heptahydrate, 0.3%-0.7% yeast extract, 0.02%-0.05% nano silica, 8-12 mg vitamin B1, and the balance being sterile water, diluted to 1000 mL, with a pH value of 5.8-6.3.

[0007] The present invention is further configured such that: the yeast extract is a highly active yeast extract prepared by low-temperature enzymatic hydrolysis, and its protein content is ≥60%.

[0008] The present invention is further configured as follows: the method for preparing the liquid spawn for increasing shiitake mushroom yield is characterized by comprising the following steps:

[0009] (1) Mix the components in the formula according to claim 1 in proportion and sonicate them at 40-50°C for 30-45 minutes to form a homogeneous culture medium;

[0010] (2) Sterilize the culture medium at 115-121℃ for 15-25 minutes and cool it to 22-26℃;

[0011] (3) Inoculate with shiitake mushroom mother culture mycelium, with an inoculation amount of 3%-5% of the culture medium volume;

[0012] (4) In a constant temperature shaker at 24-26℃ and 120-150 rpm, combine with intermittent blue light irradiation (wavelength 450-470 nm, irradiation for 2-4 hours daily) for 4-6 days until the bacterial pellet density reaches 102. 6 -10 7 CFU / mL.

[0013] The present invention is further configured such that the intensity of the blue light irradiation is 20-30 μmol·m. -2 ·s -1 And the interval between each irradiation is 2 hours.

[0014] A method for cultivating shiitake mushrooms using liquid spawn, characterized by comprising:

[0015] (1) Inoculate the liquid culture into the solid culture sticks using the pulse high pressure injection method. The injection pressure is 0.3-0.5 MPa. Each culture stick is injected at 4-6 points, and the injection volume at each point is 3-5 mL.

[0016] (2) After inoculation, the mushroom sticks are cultured in the dark at 22-25℃ and 50%-65% humidity.

[0017] The present invention is further configured such that the cultivation substrate formula of the solid mushroom sticks comprises, by weight percentage: 65%-75% broadleaf tree sawdust, 10%-15% corn cob fragments, 15%-20% wheat bran, 1%-2% shell powder, 0.5%-1% biochar, 0.1%-0.3% compound enzyme preparation, and the remainder is water, with the moisture content controlled at 58%-62%.

[0018] In summary, the present invention has the following beneficial effects:

[0019] 1. Through the synergistic effect of compound carbon and nitrogen sources and growth-promoting factors (nano silica, vitamin B1), mycelial metabolic activity is increased by 40%-50%, and the biomass accumulation rate is accelerated, solving the problem of slow growth caused by insufficient growth-promoting factors in traditional formulas.

[0020] 2. Blue light regulation combined with isothermal oscillation culture shortens the time to achieve the target bacterial density to 4-6 days (compared to the traditional 5 days), and the light intensity is 20-30 μmol·m. -2 ·s -1 Optimized parameters to avoid optical damage;

[0021] 3. Ultrasonic dissolution and high-pressure steam sterilization (115-121℃, 15-25 minutes) ensure the homogenization and sterility of the culture medium; pulsed high-pressure inoculation reduces human intervention; biochar and compound enzyme preparations are added to the solid substrate to improve the water retention and nutrient utilization of the culture medium.

[0022] 4. Digital monitoring of all parameters (temperature, humidity, light) to achieve repeatability and stability in industrial production. Detailed Implementation

[0023] To enable those skilled in the art to better understand the technical solutions of the present invention, the present invention will be further described in detail below with reference to specific embodiments. It should be noted that, in the absence of conflict, the embodiments and features in the embodiments of this application can be combined with each other.

[0024] In the description of this invention, it should be noted that the terms "upper", "lower", "inner", "outer", "top / bottom", etc. are only for the convenience of describing this invention and simplifying the description, and do not indicate or imply that the device or element referred to must have a specific orientation, or be constructed and operated in a specific orientation, and therefore should not be construed as a limitation of this invention.

[0025] In the description of this invention, it should be noted that, unless otherwise explicitly specified and limited, the terms "installed," "equipped with," "set up / connected," "connected," etc., should be interpreted broadly. For example, "connection" can be a fixed connection, a detachable connection, or an integral connection; it can be a mechanical connection or an electrical connection; it can be a direct connection or an indirect connection through an intermediate medium; it can be a connection within two components. Those skilled in the art can understand the specific meaning of the above terms in this invention based on the specific circumstances.

[0026] The present invention will now be described in detail.

[0027] A formula and method for a liquid spawn to increase shiitake mushroom yield, comprising the following components by mass percentage: 8%-12% corn flour, 5%-8% brown sugar, 2%-4% glucose, 1%-2% wheat bran, 0.4%-0.6% potassium dihydrogen phosphate, 0.15%-0.25% magnesium sulfate heptahydrate, 0.3%-0.7% yeast extract, 0.02%-0.05% nano silica, 8-12 mg vitamin B1, with the balance being sterile water, to a final volume of 1000 mL, and a pH of 5.8-6.3.

[0028] The yeast extract is a highly active yeast extract prepared by low-temperature enzymatic hydrolysis, with a protein content of ≥60%.

[0029] A method for preparing a liquid spawn to increase shiitake mushroom yield, characterized by comprising the following steps:

[0030] (1) Mix the components in the formula according to claim 1 in proportion and sonicate them at 40-50°C for 30-45 minutes to form a homogeneous culture medium;

[0031] (2) Sterilize the culture medium at 115-121℃ for 15-25 minutes and cool it to 22-26℃;

[0032] (3) Inoculate with shiitake mushroom mother culture mycelium, with an inoculation amount of 3%-5% of the culture medium volume;

[0033] (4) In a constant temperature shaker at 24-26℃ and 120-150 rpm, combine with intermittent blue light irradiation (wavelength 450-470 nm, irradiation for 2-4 hours daily) for 4-6 days until the bacterial pellet density reaches 102. 6 -10 7 CFU / mL.

[0034] The intensity of blue light irradiation is 20-30 μmol·m -2 ·s -1 And the interval between each irradiation is 2 hours.

[0035] A method for cultivating shiitake mushrooms using liquid spawn, characterized by comprising:

[0036] (1) Inoculate the liquid culture into the solid culture sticks using the pulse high pressure injection method. The injection pressure is 0.3-0.5 MPa. Each culture stick is injected at 4-6 points, and the injection volume at each point is 3-5 mL.

[0037] (2) After inoculation, the mushroom sticks are cultured in the dark at 22-25℃ and 50%-65% humidity.

[0038] The formula for solid mushroom substrate, by weight percentage, includes: 65%-75% broadleaf tree sawdust, 10%-15% corn cob fragments, 15%-20% wheat bran, 1%-2% shell powder, 0.5%-1% biochar, 0.1%-0.3% compound enzyme preparation, and the remainder is water, with a moisture content controlled at 58%-62%.

[0039] Based on traditional corn flour and brown sugar, glucose is added as a rapidly utilized carbon source, forming a complex nitrogen source with yeast extract (highly active, protein ≥60%). This provides a balanced carbon-to-nitrogen ratio for the mycelium, promoting enzyme activity and mycelial differentiation. Nano-silica enhances the mycelium's absorption efficiency of nutrients through surface adsorption. Vitamin B1, as a coenzyme precursor, accelerates mycelial metabolic cycles and shortens the lag phase. Potassium dihydrogen phosphate and magnesium sulfate heptahydrate maintain osmotic pressure and enzyme activity, ensuring stable mycelial growth.

[0040] Blue light activates the photoreceptor protein in mycelia, promoting intracellular ATP synthesis and the accumulation of secondary metabolites, while inhibiting the growth of other microorganisms. Daily irradiation for 2-4 hours with 2-hour intervals avoids photoinhibition and significantly increases mycelial density (up to 102). 6 -10 7 (CFU / mL) promotes mycelial dispersion and oxygen mass transfer through mechanical shearing force, shortening the culture cycle to 4-6 days (compared to the traditional 5 days or more), while homogenizing the size of mycelial balls and improving the viability of the strain;

[0041] By injecting liquid inoculum at multiple points (4-6 points / mushroom stick) under high pressure, the mycelium is ensured to quickly penetrate the solid substrate, reducing the risk of contamination from manual contact and increasing inoculation efficiency by more than 30%. The addition of biochar (0.5%-1%) enhances air permeability, and the compound enzyme preparation (0.1%-0.3%) decomposes lignin to release usable carbon sources. Together with shell powder, it regulates pH and creates an ideal microenvironment for mycelial colonization.

[0042] The above description is merely a preferred embodiment of the present invention, and the scope of protection of the present invention is not limited to the above embodiments. All technical solutions falling within the scope of the present invention's concept are within the scope of protection of the present invention. It should be noted that for those skilled in the art, several improvements without departing from the principle of the present invention will be readily apparent.

[0043] And refinements, these improvements and refinements should also be considered within the scope of protection of this invention.

Claims

1. A formula for a liquid spawn to increase shiitake mushroom yield, characterized in that, The product comprises the following components by weight percentage: 8%-12% corn flour, 5%-8% brown sugar, 2%-4% glucose, 1%-2% wheat bran, 0.4%-0.6% potassium dihydrogen phosphate, 0.15%-0.25% magnesium sulfate heptahydrate, 0.3%-0.7% yeast extract, 0.02%-0.05% nano silica, 8-12 mg vitamin B1, and the balance being sterile water. The volume is adjusted to 1000 mL, and the pH value is 5.8-6.

3.

2. The liquid microbial culture formulation according to claim 1, characterized in that, The yeast extract is a highly active yeast extract prepared by low-temperature enzymatic hydrolysis, with a protein content ≥60%.

3. A method for preparing a liquid spawn to increase shiitake mushroom yield, characterized in that, Includes the following steps: (1) Mix the components in the formula according to claim 1 in proportion and sonicate them at 40-50°C for 30-45 minutes to form a homogeneous culture medium; (2) Sterilize the culture medium at 115-121℃ for 15-25 minutes and cool it to 22-26℃; (3) Inoculate with shiitake mushroom mother culture mycelium, with an inoculation amount of 3%-5% of the culture medium volume; (4) In a constant temperature shaker at 24-26℃ and 120-150 rpm, combine with intermittent blue light irradiation (wavelength 450-470 nm, irradiation for 2-4 hours per day) for 4-6 days until the bacterial pellet density reaches 10. 6 -10 7 CFU / mL.

4. The preparation method according to claim 3, characterized in that, The intensity of the blue light irradiation is 20-30 μmol·m⁻²·s⁻¹, and the interval between each irradiation is 2 hours.

5. A method for cultivating shiitake mushrooms based on any one of the liquid strains described in claims 1-4, characterized in that, include: (1) Inoculate the liquid culture into the solid culture sticks using the pulse high pressure injection method. The injection pressure is 0.3-0.5 MPa. Each culture stick is injected at 4-6 points, and the injection volume at each point is 3-5 mL. (2) After inoculation, the mushroom sticks are cultured in the dark at 22-25℃ and 50%-65% humidity.

6. The cultivation method according to claim 5, characterized in that, The cultivation substrate formula of the solid mushroom sticks includes, by weight percentage: 65%-75% broadleaf tree wood chips, 10%-15% corn cob fragments, 15%-20% wheat bran, 1%-2% shell powder, 0.5%-1% biochar, 0.1%-0.3% compound enzyme preparation, and the remainder is water, with the moisture content controlled at 58%-62%.