Poria cocos liquid strain culture formula and method

Abstract

The invention discloses a poria cocos liquid strain culture formula and method. The poria cocos liquid strain culture formula comprises the following raw materials in parts by weight: 3-5 parts of soybean meal; 1-2 parts of corn flour; 1 to 1.5 parts of peptone; 8-12 parts of white sugar; 9 to 10 parts of glucose; 1 to 1.5 parts of monopotassium phosphate; 0.6 to 0.8 part of magnesium sulfate; 1000 parts of distilled water; the poria cocos liquid strain cultured by the method is short in preparation period, high in hypha activity, fast in hypha growth after inoculation, pure white in thallus, fast in feeding and high in biological conversion rate; meanwhile, the liquid strain can reduce the production and seed production cost, realizes rapid propagation and improves the production benefit.

Description

Technical Field

[0001] The invention relates to the field of biotechnology, and in particular to a formula and method for cultivating liquid tuckahoe spawn. Background Art

[0002] The original species used for the current production and cultivation of Poria cocos are generally divided into three types, namely: solid strains, liquid strains and fresh Poria cocos strains. Among them, solid strains are the production strains currently commonly used in the Poria cocos producing areas in my country. They are inoculated into solid culture bags (these bags have a variety of raw material formulas and ratios, and are sterilized at high temperature to eliminate foreign bacteria) by strain supply companies. After the mycelium grows and fills the bags (i.e., solid original species are formed), they are provided for production and application; the mycelium culture cycle of the bags is slightly longer, the strain quality between the bags varies greatly, and the production cost of the bags is relatively high.

[0003] As for fresh Poria cocos strains, producers use fresh, small-sized Poria cocos sclerotia collected this season as the production strains for planting in the next season, and then directly inoculate them into pine or basswood for production (commonly known as "meat inducement"). This strain can be sourced locally, has relatively low investment costs, and can be used to carry out next-season production in a timely manner. However, the sclerotia are easily affected by climate change and pests and diseases, and the biological transformation effect after inoculation is not ideal. Although it is an ancient way of utilizing production strains, most farmers in the main Poria cocos producing areas generally do not choose to adopt it.

[0004] Liquid strains are produced by technology-based strain supply companies, which artificially inoculate Poria cocos test tube strains (also known as "mother strains") into nutrient solution in liquid strain culture containers (the nutrient solution formula and method of culturing mycelium are the invention content of this patent) to form normal and healthy liquid strain stock mother liquor, which is then directly inoculated into bag materials for culture, production of stock strain bags, or further fermented and propagated to form a cultivation strain bag production line that can be factory-based and scaled up. Its stock strain mycelium culture cycle is short, propagation is rapid, strain quality differences between bags are small, poria formation is fast, yield is high, and the degree of mechanization is high. It is the main direction of production and development of my country's medicinal and edible fungi seed production companies in the future.

[0005] In the production process of Poria cocos liquid stock, there are too many human interferences and influencing factors such as the mother stock (also known as test tube stock), the composition, proportion, compatibility of the liquid nutrient solution, as well as the culture method, process flow and quality control, which often lead to the difficulty in ensuring the quality balance of Poria cocos liquid stock. Therefore, after the liquid stock produced by many companies is inoculated into the fungus bags, the quality of mycelium is uneven, and the biological transformation effects in production vary greatly. Therefore, there are very few domestic companies that use liquid strain production methods for Poria cocos stock, and there are almost no media reports.

[0006] In actual production, liquid strains are susceptible to infection by foreign bacteria due to large-scale inoculation, resulting in decreased vitality and degradation of species. Therefore, strict disinfection and inspection are required during the preparation of liquid stock. It is also necessary to have dedicated personnel to continuously purify and rejuvenate test tube mother strains. In addition, in the large-scale production of liquid strains, the requirements for the aseptic conditions of the surrounding environment and the skills of the operators are higher. Otherwise, it is very easy to cause significant losses in production due to contamination by pathogens, weak strain activity, and poor cultivation quality.

[0007] Among the relevant existing patents, for example: Patent CN106754428A discloses a method for liquid fermentation of Poria cocos mycelium, which has the problems of complicated steps and poor ability of the mycelium to absorb nutrients; Patent CN105385605A discloses a fast-growing Poria cocos strain and its preparation method, among which the disclosed method still has the problems of complicated preparation method and difficult to control cultivation steps.

[0008] In order to solve the above problems, a formula and method for cultivating liquid spores of Poria cocos are urgently needed. Summary of the invention

[0009] The purpose of the present invention is to provide a formula and method for culturing liquid spores of Poria cocos.

[0010] The scheme of the present invention is:

[0011] A formula for cultivating liquid tuckahoe spawn comprises the following raw materials in parts by weight:

[0012]

[0013] As a preferred technical solution, the soybean powder and corn powder are sieved through 80-100 mesh; and the distilled water is 1000 ml.

[0014] The present invention also discloses a method for preparing bacteria using a Poria cocos liquid strain culture formula, comprising the following steps:

[0015] 1) Liquid preparation

[0016] 3-5 parts by weight of soybean powder, 1-2 parts by weight of corn flour, 1-1.5 parts by weight of peptone, 8-12 parts by weight of white sugar, 9-10 parts by weight of glucose, 1-1.5 parts by weight of potassium dihydrogen phosphate and 0.6-0.8 parts by weight of magnesium sulfate are mixed with 1000 parts by weight of distilled water, dissolved, and then placed in a shaking bottle and a magnetic stirring bead;

[0017] 2) Sterilization

[0018] The shake flask was sterilized (121° C. in an autoclave for 40 minutes);

[0019] 3) Vaccination

[0020] Operate on a clean bench, take out the test tube mother culture full of Poria cocos hyphae, take out the original culture medium, select 6 to 8 pieces of mother culture as inoculated bacteria blocks, and inoculate them into the sterilized shake flask;

[0021] 4) Cultivation

[0022] The inoculated shake flask was left to stand in a natural state indoors for two days. After the standing, the shake flask was placed on a magnetic stirrer and the stirring parameters were set at 26°C and 1350 rpm for 6 to 8 hours.

[0023] Then, place the shake flask after stirring in a constant temperature shaking incubator, set the shaking parameters at 26°C and 120-150 rpm, and shake the liquid for 4-5 days;

[0024] 5) Inspection

[0025] After the shaking is completed, the quality standard of the cultivated Poria cocos liquid strain is tested, and if it meets the standard, it proceeds to the next step;

[0026] 6) Bag receiving

[0027] The shake flask liquid that meets the standards is inoculated into the bag by either artificial inoculation culture or fermentation culture.

[0028] As a preferred technical solution, there are 2 to 3 magnetic stirring beads in 1).

[0029] As a preferred technical solution, when the shake flask is sterilized in 2), the shake flask is placed in a high pressure sterilizer and sterilized at 121° C. for 40 minutes, and then cooled.

[0030] As a preferred technical solution, in the above 3), 6 to 8 pieces of 2.5×2.5 mm mother bacteria are selected as inoculated bacteria blocks.

[0031] As a preferred technical solution, the magnetic stirrer and the constant temperature shaking incubator in 4) are respectively set at 26°C; the shaking liquid is observed during 4 to 6 days, and when the bacterial balls are found to be too large, the shaking bottle is placed on the magnetic stirrer again, stirred and broken, and then the shaking liquid is carried out.

[0032] As a preferred technical solution, the quality standard of the Poria liquid spawn in 5) is: the spawn age is 5 to 7 days; the mycelium grows vigorously, is white, uniform, and dense, has obvious wall climbing phenomenon, and the mycelium can be seen in root-like chains; the spawn bottle is intact, has no odor, and is not contaminated by other bacteria.

[0033] As a preferred technical solution, the artificial inoculation in 6) is to pour the shake flask liquid into the culture medium bag in a sterile environment, with 20-30 ml of shake flask liquid poured into each bag of culture medium, and put it into the culture room for culture (culture temperature 20-22°C, air humidity <60%); the quality standard of the cultured Poria cocos culture strain is: the age of the fungus is 23-25 ​​days, the mycelium is white and dense, grows evenly, and covers the bag; milky white dew drop-shaped secretions can be seen at the tip of the mycelium, and the Poria cocos has a strong special fragrance; the bag is intact and undamaged, there is no yellow mycelium, no obvious fruiting body grows, and there is no contamination by other bacteria. The weight of the culture medium bag is 1.2-1.3 kg.

[0034] As a preferred technical solution, the fermentation culture is to expand the bacteria in the fermentation tank again, pour 800ml of shake flask liquid into a liquid strain fermentation tank with a capacity of 800-1000L under sterile conditions, and then ferment and culture for 5-7 days before inoculating into bags.

[0035] The invention adopts a formula and method for cultivating liquid tuckahoe fungus, which comprises the following raw materials in parts by weight: 5 parts of soybean powder; 2 parts of corn powder; 1.5 parts of peptone; 10 parts of white sugar; 10 parts of glucose; 1 part of potassium dihydrogen phosphate; 0.7 parts of magnesium sulfate; and 1000 parts of distilled water.

[0036] Advantages of the present invention:

[0037] The liquid strain of Poria cocos cultivated by the present invention has a short preparation cycle, strong mycelial activity, fast mycelial growth after inoculation, white bacterial body, fast material consumption, and high biological conversion rate; at the same time, the liquid strain can reduce the production cost, achieve rapid expansion and improve production efficiency. The present invention is suitable for industrial promotion, the steps are easy to control, the contamination of pathogens is reduced, the strain activity is strong, the cultivation quality is excellent, and production losses are effectively avoided. BRIEF DESCRIPTION OF THE DRAWINGS

[0038] Figure 1 The inoculated Poria cultivation bags and cells after shake flask fermentation in Example 2;

[0039] Figure 2 The Poria cocos cultivation bags are inoculated in batches after the fermentation in the fermentation tank in Example 2 is completed. DETAILED DESCRIPTION

[0040] In order to make the technical means, creative features, objectives and effects achieved by the present invention easy to understand, the present invention is further described below in conjunction with specific embodiments.

[0041] Embodiment 1:

[0042] 1) Liquid preparation:

[0043] Select 5g of 80-100 mesh sieved soybean powder, 2g of corn powder, 1.5g of peptone, 10g of white sugar, 10g of glucose, 1g of potassium dihydrogen phosphate, and 0.7g of magnesium sulfate, mix with 1000ml of distilled water and dissolve them therein, then put them into a shake flask (the total capacity of the shake flask is 1000ml, and only 800ml of liquid is needed), and put in 2-3 magnetic stirring beads.

[0044] 2) Sterilization:

[0045] The shake flask was placed in an autoclave and sterilized at 121°C for 40 minutes and then cooled.

[0046] 3) Vaccination:

[0047] Under the clean bench, take out the test tube mother culture covered with Poria cocos hyphae, remove the primordium, select 6-8 2.5×2.5 mm mother culture inoculation blocks, and inoculate them into the sterilized shake flask.

[0048] 4) Cultivation:

[0049] ① Stirring: After the inoculated shake flask is left to stand in a natural state indoors for two days, it is placed on a magnetic stirrer; the stirring parameters are set to 26°C and 1350 rpm, and stirred for 6-8 hours (in order to break up the inoculated bacteria).

[0050] ② Shake the liquid: After stirring for 6-8 hours, place the shake bottle in a constant temperature shaking incubator, set the shaking parameters at 26°C and 120-150 rpm, and shake the liquid for 4-5 days.

[0051] 5) Observation

[0052] During the shaking process, if the bacterial balls are found to be too large, the shaking bottle should be placed back on the magnetic stirrer for stirring and breaking.

[0053] 6) Inspection

[0054] The quality standard of cultivated Poria cocos liquid culture is: the culture age is 5-7 days; the mycelium grows vigorously, is white, uniform and dense, has obvious wall climbing phenomenon, and the mycelium can be seen in root-like chain. The bottle of the culture is intact, without odor and without contamination by other bacteria.

[0055] 7) Bag filling: If artificial inoculation is selected, the shake bottle liquid is poured into the culture medium bag in a sterile environment, and each bag of culture medium weighs 1.2-1.3kg (the amount poured is 20-30ml), and the culture is placed in a culture room (culture temperature 20-22℃, air humidity <60%). The quality standards of the cultured Poria cocos culture strain are: the age of the culture is 23-25 ​​days, the mycelium is white and dense, grows evenly, and covers the entire bag; milky white dew drop-like secretions can be seen at the tip of the mycelium, and the Poria cocos has a strong special fragrance; the bag is intact and undamaged, there is no yellow mycelium, no obvious fruiting body grows, and there is no contamination by other bacteria.

[0056] Embodiment 2:

[0057] 1) Liquid preparation:

[0058] Select 3g of 80-100 mesh sieved soybean powder, 1.5g of corn flour, 1g of peptone, 12g of sugar, 10g of glucose, 1.5g of potassium dihydrogen phosphate, and 0.6g of magnesium sulfate, mix with 1000ml of distilled water and dissolve them therein, and add 2-3 magnetic stirring beads; then, put them into a shake flask (the total capacity of the shake flask is 1000ml, and only 800ml of liquid is needed).

[0059] 2) Sterilization:

[0060] The shake flask was placed in an autoclave and sterilized at 121°C for 40 minutes.

[0061] 4) Vaccination:

[0062] Under the clean bench, take out the test tube mother culture covered with Poria cocos hyphae, remove the primordium, select 6-8 2.5×2.5 mm mother culture inoculation blocks, and inoculate them into the sterilized shake flask.

[0063] 4) Cultivation:

[0064] ① Stirring: After the inoculated shake flask is left to stand in a natural state indoors for two days, it is placed on a magnetic stirrer; the stirring parameters are set at 26°C and 1350 rpm, and stirred for 6-8 hours (in order to break up the inoculated bacteria);

[0065] ② Shake the liquid: After stirring for 6-8 hours, place the shake bottle in a constant temperature shaking incubator, set the shaking parameters at 26°C and 120-150 rpm, and shake the liquid for 4-5 days.

[0066] 5) Observation

[0067] During the shaking process, if the bacterial balls are found to be too large, the shaking bottle should be placed back on the magnetic stirrer for stirring and breaking.

[0068] 6) Inspection

[0069] The quality standard of cultivated Poria cocos liquid culture is: the culture age is 5-7 days; the mycelium grows vigorously, is white, uniform and dense, has obvious wall climbing phenomenon, and the mycelium can be seen in root-like chain. The bottle of the culture is intact, without odor and without contamination by other bacteria.

[0070] 7) Bag inoculation: If artificial inoculation is selected, pour the shake bottle liquid into the culture medium bag in a sterile environment, 20-30 ml per bag (material weight 1.2-1.3 kg), and place it in a culture room for culture (culture temperature 20-22°C, air humidity <60%). The quality standards of the cultured Poria cocos culture strain are: 23-25 ​​days old, white and dense mycelium, uniform growth, covering the bag; milky white dew drop-like secretions can be seen at the tip of the mycelium, and the Poria cocos-specific fragrance is strong; the bag is intact and undamaged, there is no yellow mycelium, no obvious fruiting body growth, and no contamination by other bacteria.

[0071] Embodiment 3:

[0072] 1) Liquid preparation:

[0073] Select 4g of 80-100 mesh sieved soybean powder, 1g of corn powder, 1.5g of peptone, 10g of sugar, 10g of glucose, 1g of potassium dihydrogen phosphate, and 0.8g of magnesium sulfate, mix with 1000ml of distilled water and dissolve them therein, and add 2-3 magnetic stirring beads; then, put them into a shake flask (the total capacity of the shake flask is 1000ml, and only 800ml of liquid is needed).

[0074] 2) Sterilization:

[0075] The shake flask was placed in an autoclave and sterilized at 121°C for 40 minutes.

[0076] 5) Vaccination:

[0077] Under the clean bench, take out the test tube mother culture covered with Poria cocos hyphae, remove the primordium, select 6-8 2.5×2.5 mm mother culture inoculation blocks, and inoculate them into the sterilized shake flask.

[0078] 4) Cultivation:

[0079] ① Stirring: After the inoculated shake flask is left to stand in a natural state indoors for two days, it is placed on a magnetic stirrer; the stirring parameters are set at 26°C and 1350 rpm, and stirred for 6-8 hours (in order to break up the inoculated bacteria);

[0080] ② Shake the liquid: After stirring for 6-8 hours, place the shake bottle in a constant temperature shaking incubator, set the shaking parameters at 26°C and 120-150 rpm, and shake the liquid for 4-5 days.

[0081] 5) Observation

[0082] During the shaking process, if the bacterial balls are found to be too large, the shaking bottle should be placed back on the magnetic stirrer for stirring and breaking.

[0083] 6) Inspection

[0084] The quality standard of cultivated Poria cocos liquid culture is: the culture age is 5-7 days; the mycelium grows vigorously, is white, uniform and dense, has obvious wall climbing phenomenon, and the mycelium can be seen in root-like chain. The bottle of the culture is intact, without odor and without contamination by other bacteria.

[0085] 7) Bag inoculation: If artificial inoculation is selected, pour the shake bottle liquid into the culture medium bag in a sterile environment, 20-30 ml per bag (material weight 1.2-1.3 kg), and place it in a culture room for culture (culture temperature 20-22°C, air humidity <60%). The quality standards of the cultured Poria cocos culture strain are: 23-25 ​​days old, white and dense mycelium, uniform growth, covering the bag; milky white dew drop-like secretions can be seen at the tip of the mycelium, and the Poria cocos-specific fragrance is strong; the bag is intact and undamaged, there is no yellow mycelium, no obvious fruiting body growth, and no contamination by other bacteria.

[0086] Embodiment 4:

[0087] 1) Liquid preparation:

[0088] Select 5g of 80-100 mesh sieved soybean powder, 2g of corn powder, 1.5g of peptone, 10g of sugar, 10g of glucose, 1g of potassium dihydrogen phosphate, and 0.7g of magnesium sulfate, mix with 1000ml of distilled water and dissolve them therein, and add 2-3 magnetic stirring beads; then, put them into a shake flask (the total capacity of the shake flask is 1000ml, and only 800ml of liquid is needed).

[0089] 2) Sterilization:

[0090] The shake flask was placed in an autoclave and sterilized at 121°C for 40 minutes.

[0091] 3) Vaccination:

[0092] Under the clean bench, take out the test tube mother culture full of Poria cocos hyphae, remove the primordium, select 6-8 pieces of 2.5×2.5 mm mother culture as inoculation blocks, and inoculate them into the sterilized shake flask.

[0093] 4) Cultivation:

[0094] ① Stirring: After the inoculated shake flask is left to stand in a natural state indoors for two days, it is placed on a magnetic stirrer; the stirring parameters are set at 26°C and 1350 rpm, and stirred for 6-8 hours (in order to break up the inoculated bacteria);

[0095] ② Shake the liquid: After stirring for 6-8 hours, place the shake bottle in a constant temperature shaking incubator, set the shaking parameters at 26°C and 120-150 rpm, and shake the liquid for 4-5 days;

[0096] 5) Observation

[0097] During the shaking process, if the bacterial balls are found to be too large, the shaking bottle should be placed back on the magnetic stirrer for stirring and breaking.

[0098] 6) Inspection

[0099] The quality standard of cultivated Poria cocos liquid culture is: the culture age is 5-7 days; the mycelium grows vigorously, is white, uniform and dense, has obvious wall climbing phenomenon, and the mycelium can be seen in root-like chain. The bottle of the culture is intact, without odor and without contamination by other bacteria.

[0100] Specific quality standards for Poria cocos liquid strains:

[0101] a. Quality standard of Poria mother culture: The age of the culture is 7 to 10 days; the mycelium is white, uniform, dense and strong, with a strong Poria-specific aroma; crystal clear dew-drop-like secretions can be seen on the surface of the mycelium; the culture tube is intact, the cotton plug is tight, and there is no contamination from other bacteria.

[0102] b. Quality standard of original Poria cocos strain: The strain age is 15 to 30 days; the mycelium grows vigorously, is white, uniform, and dense, and the wall climbing phenomenon is obvious, and root-like cords can be seen. Milky white dew drop-like secretions can be seen at the tip of the mycelium, and the Poria cocos has a strong aroma; the strain bottle is intact and free of contamination by other bacteria.

[0103] C. Quality standard for Poria cultivation strains: The fungus age is 30 to 45 days; the mycelium is white and dense, grows evenly, and covers the entire bag; milky white dew-drop-like secretions can be seen at the tip of the mycelium, and the Poria has a strong and unique aroma; the bag is intact and undamaged, with no yellowed mycelium, no fruiting bodies growing, and no contamination from other bacteria.

[0104] 7) Bag transfer: Use a fermentation tank to expand the culture again. Pour 800ml of shake flask liquid into a 800-1000L liquid culture fermentation tank under sterile conditions. Then, ferment and culture for another 5-7 days before inoculating into the bag.

[0105] Yunnan Institute of Traditional Chinese Medicine, Yunnan Junma Biotechnology Co., Ltd., and Jinggu Dai and Yi Autonomous County Edible Fungi Research Institute use the liquid tuckahoe strain prepared and produced by the present invention, which can be directly used for production in automated inoculators (each 800ml shake bottle mother strain can inoculate 800-1000L of liquid fermentation tanks, and the fermentation period is only about 5-7 days; each fermentation tank can be inoculated with 8000-10000 tuckahoe cultivation bags). Since April 2023, more than 60,000 bags of tuckahoe liquid strain cultivation bags have been produced.

[0106] The above shows and describes the basic principles, main features and advantages of the present invention. It should be understood by those skilled in the art that the present invention is not limited to the above embodiments. The above embodiments and descriptions are only for explaining the principles of the present invention. Without departing from the spirit and scope of the present invention, the present invention may have various changes and improvements, which fall within the scope of the present invention. The scope of protection of the present invention is defined by the attached claims and their equivalents.

Claims

1. A formula for cultivating liquid spores of Poria cocos, characterized in that: The invention comprises the following raw materials in parts by weight:

2. A Poria cocos liquid spawn culture formula as claimed in claim 1, characterized in that: The soybean flour and corn flour are sieved through 80-100 meshes.

3. A method for preparing bacteria using the Poria cocos liquid spawn culture formula as claimed in claim 1, characterized in that: The following steps are involved: 1) Liquid preparation 3-5 parts by weight of soybean powder, 1-2 parts by weight of corn flour, 1-1.5 parts by weight of peptone, 8-12 parts by weight of white sugar, 9-10 parts by weight of glucose, 1-1.5 parts by weight of potassium dihydrogen phosphate and 0.6-0.8 parts by weight of magnesium sulfate are mixed with 1000 parts by weight of distilled water, dissolved, and then placed in a shaking bottle and a magnetic stirring bead; 2) Sterilization Sterilize the shake flask by high temperature treatment; 3) Vaccination Operate on a clean bench, take out the test tube mother culture full of Poria cocos hyphae, take out the original culture medium, select 6 to 8 pieces of mother culture as inoculated bacteria blocks, and inoculate them into the sterilized shake flask; 4) Cultivation The inoculated shake flask was left to stand in a natural state indoors for two days. After the standing, the shake flask was placed on a magnetic stirrer at a stirring parameter of 1350 rpm for 6 to 8 hours. Place the shake flask after stirring in a constant temperature shaking incubator with a shaking parameter of 120 to 150 rpm for 4 to 5 days; 5) Inspection After the shaking liquid treatment is completed, the quality standard of the cultivated Poria cocos liquid strain is tested, and if it meets the standard, it proceeds to the next step; 6) Bag receiving The shake flask liquid that meets the standards is inoculated into the bag by either artificial inoculation culture or fermentation culture.

4. A method for preparing bacteria using a Poria cocos liquid spawn culture formula as claimed in claim 3, characterized in that: In the above 1), there are 2 to 3 magnetic stirring beads.

5. A method for preparing bacteria using a liquid spawn culture formula of Poria cocos as claimed in claim 3, characterized in that: When the shaking flask is sterilized in 2), the shaking flask is placed in a high pressure sterilizer and sterilized at 121° C. for 40 minutes.

6. A method for preparing bacteria using a Poria cocos liquid spawn culture formula as claimed in claim 3, characterized in that: In the above 3), 6 to 8 pieces of 2.5×2.5 mm mother bacteria are selected as inoculated bacteria blocks.

7. A method for preparing bacteria using a Poria cocos liquid spawn culture formula as claimed in claim 3, characterized in that: In the above 4), the magnetic stirrer and the constant temperature shaking incubator are respectively set at 26° C.; the shaking liquid is observed during 4 to 6 days. When the bacterial balls are found to be too large, the shaking bottle is placed on the magnetic stirrer again, stirred and broken, and then the shaking liquid is performed.

8. A method for preparing bacteria using a Poria cocos liquid spawn culture formula as claimed in claim 3, characterized in that: The quality standard of the Poria liquid culture in 5) is that the culture age is 5 to 7 days; the mycelium grows vigorously, is white, uniform, dense, has obvious wall climbing phenomenon, and the mycelium can be seen in root-like chains; the bottle of the culture is intact, has no odor, and is not contaminated by other bacteria.

9. A method for preparing bacteria using a liquid spawn culture formula of Poria cocos as claimed in claim 3, characterized in that: The artificial inoculation in 6) is to pour the shake flask liquid into the culture medium bag in a sterile environment, with the amount of shake flask liquid poured into each culture medium bag being 20-30 ml, and then put it into the culture room for cultivation at a temperature of 22-24° C. and an air humidity of 60-75%. The quality standards of the cultured Poria cocos culture strains are: the age of the culture is 23-25 ​​days, the mycelium is white and dense, grows evenly, and covers the entire bag; milky white dew drop-like secretions can be seen at the tip of the mycelium, and the Poria cocos has a strong aroma. The fungus bag is intact and undamaged, has no yellowed hyphae, no obvious fruiting bodies, and is not contaminated by other bacteria.

10. A method for preparing bacteria using a Poria cocos liquid spawn culture formula as claimed in claim 3, characterized in that: The fermentation culture is to expand and culture the bacteria in the fermentation tank again. 800 ml of the shake bottle liquid is poured into a liquid strain fermentation tank with a capacity of 800-1000 L under sterile conditions. Then, it is fermented and cultured for 5-7 days before being inoculated into bags.

Images