Grifola frondosus liquid strain and method for cultivating grifola frondosus by using liquid strain

[0019] Embodiment one:

[0020] Production of parent species: Grifola frondosa strain block (preserved by the strain preservation center of the Institute of Edible Fungi, Shanghai Academy of Agricultural Sciences, code: GF1) was inoculated in PDA medium, and cultured in the dark at 25°C for 10 days;

[0021] First-class liquid strain production (shake flask culture method): culture solution formula: 5% potato, 2% glucose, 3% corn flour, 1% bran extract, 0.8% agar, natural pH; after the culture solution is prepared , put into a 500mL Erlenmeyer flask, the volume of each bottle is 100mL, and add 10 glass beads, add a cotton plug and then wrap it with kraft paper to seal, after sterilization, cool down to below 25°C, put it in and smash it with a homogenizer The mother species of , were cultured statically at 25°C for 48 hours, and then placed on a shaker for shaking culture at a frequency of 100r/min. After cultivating for 10 days, it can be seen that the culture solution is clear and transparent, and a large number of mycelia balls of Grifola frondosa are suspended in the solution;

[0022] Production of secondary liquid strains (i.e. liquid strains) (liquid fermentation tank cultivation): the formula of the culture solution is the same as above, after the culture solution is prepared, put it into the fermentation tank, the volume of each bottle is 3L, the ventilation equipment of the culture solution and the fermentation tank Sterilize at the same time, when it is cooled to below 25°C after sterilization, insert 200ml of the first-grade liquid bacteria, open the air intake device and continuously replenish the filtered fresh air, after 14 days of cultivation, it can be seen that the culture medium is clear and transparent, and a large amount of ash is suspended in the liquid Tree flower mycelium ball; get liquid seed.

[0023] Production of Grifola frondosa Cultivation Fungus Bag: The ratio of medium raw materials is: 30% wood chips, 30% cottonseed hulls, 20% bran, 5% corn flour, 1% brown sugar, 1% gypsum, and 13% soil. Add water and stir evenly so that the water content is 65%. Pack it into bags with an automatic bagging machine. Each bag contains 900g of wet material and a height of 14cm. After sterilization, each bag receives 20ml of liquid bacteria, and then placed at a temperature of 22 ° C, a relative humidity of 60%, CO 2 Concentration of 1000ppm culture, 28d mycelium is covered with bacterial bag;

[0024] Mushroom cultivation management: After the mycelium is full of bags, it needs 10 days of after-ripening. After the mycelium has penetrated completely, cut a hole on the side. Place them in a dark room at a temperature of 22°C and a relative humidity of 95% to stimulate bud emergence. After 7 days, a spherical primordium appeared at the opening. At this time, set the temperature at 19°C, relative humidity at 95%, and CO 2 The concentration is 500ppm, and the indoor fluorescent lamp is turned on. The spherical primordium gradually differentiates into a brain shape, and it can be harvested when the fruiting body grows until the cap is fan-shaped, and the pore just appears.

[0025] Embodiment two:

[0026] The parent species is made the same as in Example 1; the strain used is GF2 (preserved by the Strain Collection Center of the Institute of Edible Fungi, Shanghai Academy of Agricultural Sciences, number: GF2)

[0027] First-class liquid strain system (shake flask culture method): culture solution formula: 5% potato, 2% glucose, 2% corn flour, 3% bran extract, 1% agar, natural pH. Bottling, sterilization, and inoculation were the same as in Example 1. After inoculation, they were placed on a shaking table for shaking culture, and the shaking frequency was 150 r/min. The room temperature of the shaker was controlled at 24°C. After 9 days of cultivation, the culture liquid was clear and transparent, and a large number of mycelia balls of Grifola frondosa were suspended in the liquid;

[0028] Production of secondary liquid strains (cultivation in liquid fermenters): the formulation of the culture medium, bottling, and sterilization are the same as in Example 1. After sterilization, when cooling to below 25°C, add 150ml of primary liquid strains and open the air intake The device continuously replenishes the fresh air that has been filtered. After 13 days of cultivation, it can be seen that the culture liquid is clear and transparent, and there are a large number of mycelial balls of Grifola frondosa suspended in the liquid, which is the liquid strain;

[0029] Grifola frondosa cultivation fungus bag making: the ratio of medium raw materials is the same as that in Example 1, and it is packed by an automatic bagging machine, with 800 g of wet material in each bag and a height of 14 cm. After sterilization, each bag received 25ml of liquid bacteria, placed at a temperature of 23°C, a relative humidity of 65%, CO 2 The concentration is 1000ppm culture, 30d mycelium is covered with bacterial bag;

[0030] Mushroom cultivation management: the method of cutting the mushroom bag is the same as in Example 1, and then put it in a dark room at a temperature of 22°C and a relative humidity of 98% to stimulate bud emergence. After 7 days, a spherical primordium appeared at the opening. At this time, set the temperature at 19°C, relative humidity at 93%, and CO 2 The concentration is 800ppm, and the indoor fluorescent lamp is turned on. The spherical primordium gradually differentiates into a brain shape, and it can be harvested when the fruiting body grows until the cap is fan-shaped, and the pore just appears.

[0031] Compared with the cultivation method utilizing solid strains, the present invention has the following advantages:

[0032] Compared with solid strains, the production cycle of liquid strains is shorter, the age of the strains is consistent and the cost is low. After inoculation, it is dispersed to each part of the fungus bag, and the mycelium balls germinate at the same time, which makes the germination time of the fungus bag short and the age of the bacteria is consistent, which is convenient for the later management of mushroom production. The liquid bacteria cover the surface of the bacteria bag, and the rapid germination of the bacteria greatly reduces the pollution rate of the bacteria bag. Compared with the use of solid strains, the proportion of bacteria bag primordia formed by liquid strains was higher, and the yield and quality of Grifola frondosa were high.