Production method and application method of reduced liquid strain of edible fungi

Abstract

The invention discloses an edible fungus reduced liquid strain production method which comprises the following steps: S11, primary culture: inoculating a mycelium slant into a liquid culture medium of a conical flask, and carrying out shaking culture at 25-30 DEG C and 100-200rpm, so that the conical flask is filled with mycelium pellets with uniform size; s12, multiplication culture: transferring the mycelium pellets into a liquid culture medium of a new conical flask according to a volume ratio of 10-20%, and carrying out standing culture at 25-30 DEG C for 7-10 days; s13, thallus concentration: after amplification culture is finished, standing to remove supernate, and collecting a lower-layer mycelium pellet concentrated solution; and S14, vacuum packaging: performing vacuum packaging on the mycelium pellet concentrated solution under a sterile condition, and storing at 4 DEG C for later use. The inoculation proportion of the reduced liquid strain is 8-15%. The reduced liquid strain disclosed by the invention is good in stability, can be preserved for 2-3 months under the condition of refrigeration at 4 DEG C, and is relatively good in activity; by utilizing the character, the preparation of the reduced strain can be completed through research institutes or production enterprises, and then the reduced strain is directly supplied to vast farmer households, so that not only can the production period of the produced strain bag be shortened, but also the edible fungus cultivation mode which is widely used at present can be well linked.

Description

Technical Field

[0001] The present invention relates to the field of bioengineering technology, and in particular to a production method for culturing edible fungi liquid strains by a reduction method. The present invention also provides an application method for improving the reduced edible fungi liquid strains. Background Art

[0002] Liquid spawn cultivation technology for edible fungi represents an interdisciplinary field, encompassing both industrial fermentation technology and the edible fungi industry. Because the liquid environment is rich in nutrients, controlled environmental conditions promote vigorous mycelial metabolism, resulting in rapid mycelial growth and reproduction, allowing for the production of large quantities of mycelial pellets in a short period of time. The growth cycle of liquid spawn is typically 5-7 days, while traditional solid spawn typically takes 30-50 days to reach full growth. Furthermore, liquid spawn offers high consistency, with consistent mycelial growth and development, high vitality, and rapid development. Following inoculation, multiple germination points are observed, making management easier. The consistent appearance of buds and fruiting times facilitates harvesting and processing. Furthermore, the fluid nature of liquid spawn facilitates mechanization and automation of inoculation operations, effectively improving efficiency.

[0003] However, liquid spawn also has significant limitations. Liquid fermentation equipment for edible fungi is expensive, and its operation requires extensive expertise, making it difficult for ordinary farmers to master. Therefore, liquid spawn is currently only used by edible fungi production companies and is difficult to promote among the general farming community. Summary of the Invention

[0004] The present invention aims to provide a method for producing reduced liquid spawn of edible fungi. The reduced liquid spawn obtained by this production method can be directly supplied to a large number of growers, thereby shortening the production cycle of mushroom bags and effectively integrating with the currently widely used edible fungi cultivation model. The present invention also provides a method for using the reduced liquid spawn.

[0005] In order to achieve the above technical purpose, the technical solution of the present invention is:

[0006] A method for producing reduced liquid spawn of edible fungi comprises the following steps:

[0007] S11 Primary Culture: Inoculate a mycelial slant into liquid culture in an Erlenmeyer flask. Incubate at 25-30°C, 100-200 rpm, and shake for 5-7 days until the flask is filled with uniformly sized mycelial pellets. The liquid culture medium contains 8-12 g / L peptone, 3-7 g / L yeast extract, and 3-7 g / L sodium chloride, with a natural pH.

[0008] S12 expansion culture: transfer the mycelial pellets to the liquid culture medium in a new conical flask at a volume ratio of 10-20%, and culture at 25-30°C for 7-10 days; during the expansion culture, add appropriate amount of defoaming agent and vitamin B1 to promote the growth of the mycelial pellets.

[0009] S13 bacterial concentration: After the expansion culture is completed, let it stand for 12 hours, remove the supernatant, and collect the concentrated liquid of the lower mycelial ball;

[0010] S14 Vacuum packaging: Under sterile conditions, vacuum-pack the mycelial pellet concentrate and store it at 4°C for later use.

[0011] A method for applying a reduced liquid bacterial strain comprises the following steps:

[0012] S21: Add the concentrated mycelial pellets to sterile physiological saline at a volume ratio of 1:1, mix well, and then inoculate at an inoculation ratio of 8-15%.

[0013] The reduced liquid strain of the present invention has good stability and can be preserved for 2-3 months under refrigerated conditions at 4°C with good activity. Utilizing this property, the reduced strain can be prepared by research institutes or production enterprises and then directly supplied to a large number of growers, which can not only shorten the production cycle of the mushroom bags, but also well connect with the currently widely used edible fungus cultivation model. DETAILED DESCRIPTION

[0014] Example 1

[0015] The liquid culture medium contained 8 g / L peptone, 7 g / L yeast extract, 5 g / L sodium chloride, and natural pH.

[0016] A method for producing reduced liquid spawn of edible fungi comprises the following steps:

[0017] S11 Primary Culture: Add liquid culture medium to a conical flask, take a 2cm×2cm small piece of slant culture medium covered with Ganoderma lucidum mycelium and inoculate it into the liquid culture medium, inoculating 3 pieces per conical flask. Incubate at 27℃ and 150rpm with shaking for 5-7 days, so that the conical flask is filled with mycelial balls of uniform size.

[0018] S12 expansion culture: Take a new Erlenmeyer flask and add liquid culture medium to it. Inoculate the mycelial pellets obtained from the primary culture into the new Erlenmeyer flask at a volume ratio of mycelial pellets to liquid culture medium of 1:9. Incubate at 25°C for 10 days.

[0019] S13 bacterial concentration: After the expansion culture is completed, the culture is allowed to stand for 12 hours, the supernatant is removed through a pipe, and the concentrated liquid of the lower mycelial ball is collected;

[0020] S14 Vacuum packaging: Under sterile conditions, the mycelial pellet concentrate is placed in a bag and vacuum-sealed. After vacuum packaging, it is stored at an ambient temperature of 4°C for future use.

[0021] Example 2

[0022] The liquid culture medium contained 12 g / L peptone, 5 g / L yeast extract, 7 g / L sodium chloride, and natural pH.

[0023] A method for producing reduced liquid spawn of edible fungi comprises the following steps:

[0024] S11 Primary Culture: Add liquid culture medium to a conical flask, take a 2cm×2cm small piece of slant culture medium covered with Ganoderma lucidum mycelium and inoculate it into the liquid culture medium, inoculating 4 pieces per conical flask. Culture at 25℃ and 200rpm with shaking for 5-7 days, so that the conical flask is filled with mycelial balls of uniform size.

[0025] S12 expansion culture: Take a new Erlenmeyer flask, add liquid culture medium to it, and add an appropriate amount of vitamin B1. Inoculate the mycelial pellets obtained from the primary culture into the new Erlenmeyer flask at a volume ratio of mycelial pellets to liquid culture medium of 1.6:8. Incubate at 28°C for 7 days.

[0026] S13 bacterial concentration: After the expansion culture is completed, let it stand for 12 hours, remove the supernatant using a pipe, and collect the concentrated liquid of the lower mycelial ball;

[0027] S14 Vacuum packaging: Under sterile conditions, the mycelial pellet concentrate is placed in a bag and vacuum-sealed. After vacuum packaging, it is stored at an ambient temperature of 4°C for future use.

[0028] Example 3

[0029] The liquid culture medium contained 10 g / L peptone, 3 g / L yeast extract, 3 g / L sodium chloride, and natural pH.

[0030] A method for producing reduced liquid spawn of edible fungi comprises the following steps:

[0031] S11 Primary Culture: Add liquid culture medium to a conical flask, take a 2cm×2cm small piece of slant culture medium covered with Ganoderma lucidum mycelium and inoculate it into the liquid culture medium, inoculating 3 pieces per conical flask. Incubate at 30℃ and 100 rpm with shaking for 5-7 days, so that the conical flask is filled with mycelial balls of uniform size.

[0032] S12 expansion culture: Take a new conical flask, add liquid culture medium into the conical flask, and add appropriate amount of defoaming agent and vitamin B 11 The mycelial pellets obtained from the primary culture were inoculated into a new conical flask with a volume ratio of mycelial pellets to liquid culture medium of 1:8 and cultured at 30°C for 7 days.

[0033] S13 bacterial concentration: After the expansion culture is completed, the culture is allowed to stand for 12 hours, the supernatant is removed through a pipe, and the concentrated liquid of the lower mycelial ball is collected;

[0034] S14 Vacuum packaging: Under sterile conditions, the mycelial pellet concentrate is placed in a bag and vacuum-sealed. After vacuum packaging, it is stored at an ambient temperature of 4°C for future use.

[0035] Example 4

[0036] A method for applying a reduced liquid strain of Ganoderma lucidum comprises the following steps:

[0037] The formula of the Ganoderma lucidum culture bag is: 98% wheat or barley, 2% calcium carbonate, the moisture content of the culture medium is 55%-60%, and the wet weight of each bag is 1 kg. Ordinary pressure sterilization requires maintaining 100℃ for 8-12 hours, and high pressure sterilization requires maintaining 121℃ for 2-3 hours.

[0038] The mycelial pellet concentrate prepared in Example 1 was added to sterile physiological saline at a volume ratio of 1:1 and mixed evenly. The mixture was then inoculated into a Ganoderma lucidum culture bag at an inoculation ratio of 8%. The culture was carried out in the dark at 25°C. The mycelium filled the bag for 30 days. The mycelium growth rate was 10% higher than that of using solid spawn.

[0039] Example 5

[0040] A method for applying a reduced liquid strain of Ganoderma lucidum comprises the following steps:

[0041] The formula of the Ganoderma lucidum culture bag is: 98% wheat or barley, 2% calcium carbonate, the moisture content of the culture medium is 55%-60%, and the wet weight of each bag is 1 kg. Ordinary pressure sterilization requires maintaining 100℃ for 8-12 hours, and high pressure sterilization requires maintaining 121℃ for 2-3 hours.

[0042] The mycelial pellet concentrate prepared in Example 2 was added to sterile physiological saline at a volume ratio of 1:1 and mixed evenly. The mixture was then inoculated into a Ganoderma lucidum culture bag at an inoculation ratio of 10%. The culture was carried out in the dark at 25°C. The mycelium filled the bag for 28 days, and the mycelium growth rate was increased by 12% compared with that using solid strains.

[0043] Example 6

[0044] A method for applying a reduced liquid strain of Ganoderma lucidum comprises the following steps:

[0045] The formula of the Ganoderma lucidum culture bag is: 98% wheat or barley, 2% calcium carbonate, the moisture content of the culture medium is 55%-60%, and the wet weight of each bag is 1 kg. Ordinary pressure sterilization requires maintaining 100℃ for 8-12 hours, and high pressure sterilization requires maintaining 121℃ for 2-3 hours.

[0046] The mycelial pellet concentrate prepared in Example 3 was added to sterile physiological saline at a volume ratio of 1:1 and mixed evenly. The mixture was then inoculated into a Ganoderma lucidum culture bag at an inoculation ratio of 15%. The culture was carried out in the dark at 25°C. The mycelium filled the bag for 25 days. The mycelium growth rate was 16% higher than that of using solid spawn.

[0047] The above embodiments do not limit the present invention in any way, and any technical solutions obtained by equivalent replacement or equivalent transformation fall within the protection scope of the present invention.

Claims

1. A method for producing reduced liquid spawn of edible fungi, characterized in that: The following steps are involved: S11 Primary Culture: Inoculate the mycelium slant into the liquid culture medium in an Erlenmeyer flask and culture at 25-30°C and 100-200 rpm with shaking until the flask is filled with mycelial balls of uniform size. S12 expansion culture: transfer the mycelial pellets to the liquid culture medium in a new conical flask at a volume ratio of 10-20%, and culture at 25-30℃ for 7-10 days; S13 bacterial concentration: After the expansion culture is completed, the supernatant is removed and the concentrated liquid of the mycelial balls in the lower layer is collected; S14 Vacuum packaging: Under sterile conditions, vacuum-pack the mycelial pellet concentrate and store it at 4°C for later use.

2. The method for producing a reduced liquid spawn of edible fungi according to claim 1, wherein: The liquid culture medium in step S11 contains 8-12 g / L of peptone, 3-7 g / L of yeast extract, 3-7 g / L of sodium chloride, and has a natural pH.

3. The method for producing a reduced liquid spawn of edible fungi according to claim 1, wherein: The liquid culture medium in step S12 contains 8-12 g / L of peptone, 3-7 g / L of yeast extract, 3-7 g / L of sodium chloride, and has a natural pH.

4. The method for producing a reduced liquid spawn of edible fungi according to claim 1, wherein: The liquid culture medium in step S12 is added with a defoaming agent and / or vitamin B1.

5. A method for applying a reduced liquid bacterial strain, characterized in that: The mycelial pellet concentrate obtained by the method for producing the reduced liquid spawn of edible fungi according to any one of claims 1 to 4 is added to sterile physiological saline at a volume ratio of 1:1, mixed evenly, and then inoculated at an inoculation ratio of 8-15%.