TW201742918A - A cultivation method of North Cordyceps sinensis - Google Patents

Abstract

A cultivation method of North Cordyceps sinensis, comprises the following steps: step 1: purifying bacteria; step 2: producing bacteria; step 3: producing nutrient solution; step 4 producing solid culture material; step 5: cultivating mycelia; step 6: prompting color transfer of the mycelial; step 7: completing the cultivation of Cordyceps militaris. By following these steps, the simple and stable mass production of Cordyceps militaris can be guaranteed, which is an cost-effective method for cultivating Cordyceps militaris.

Description

Method for cultivating Cordyceps militaris

The invention mainly relates to a cultivation technique of Cordyceps sinensis, in particular to a method for producing a Cordyceps militaris fruit body by a manual method for a Cordyceps militaris.

Cordyceps sinensis, also known as summer grass worm, referred to as Cordyceps, is a valuable Chinese herbal medicine and enjoys a high reputation at home and abroad. Cordyceps has the effects of lowering blood sugar, anti-tumor, lowering blood fat, enhancing immunity, enhancing sexual function, replenishing kidney and strengthening yang, benefiting essence, preventing aging and prolonging life. It can improve the detoxification ability of the liver and play a role in protecting the liver. Cordyceps contains 19 essential amino acids and abundant protein and a large number of vitamins. It contains more than 17 kinds of trace elements necessary for the human body. Among them, the "selenium" which is called the king of anti-cancer is extremely rich. It also contains a large amount of superoxide dismutase (SOD), which is a super substance that protects human body cells, helping to eliminate free radicals produced by excessive cell mitochondria and preventing cells from being oxidized, aged or destroyed. It can enhance the activity of SOD in the red blood cells of the body, activate cells, and resist aging and anti-oxidation.

Cordyceps militaris is of the same genus and different species as Cordyceps sinensis. The northern savannah is produced by Changbai Mountain and Cordyceps sinensis. Cordyceps militaris is also known as Cordyceps militaris, Cordyceps militaris or Golden Cordyceps sinensis (since the color is golden yellow, hence the name). The value of medicinal and medicinal properties is comparable to that of traditional Cordyceps sinensis. The content of cordycepin in artificially cultivated Cordyceps militaris is several times higher than that of wild Cordyceps sinensis. It has the functions of nourishing the lungs and tonifying the kidney, stopping bleeding and removing phlegm, expanding the trachea, calming, antibacterial and lowering blood pressure. Cordycepin can be used for anti-cancer and anti-virus.

As the natural ecology continues to be destroyed, the wild resources of Cordyceps sinensis are scarce and prices have been rising. In order to meet the market demand, many research institutes at home and abroad have carried out research on artificial cultivation of Cordyceps sinensis. However, because Cordyceps sinensis is a fungus that is parasitic on the larvae of the bat moth, one of the host insects lives in the plateau and is difficult to obtain. The formation mechanism and formation conditions of the two bacteria and insects are difficult to determine. Therefore, there is currently no technical ability to cultivate Cordyceps sinensis, but Cordyceps militaris can only be cultivated by artificial synthetic medium to obtain its sub-seat, or the hyphae of Cordyceps can be obtained by liquid culture. Body, cultivate a substitute similar to natural Cordyceps, alleviate the contradiction between supply and demand in the market.

The main object of the present invention is to provide a simple and cost-effective method for cultivating Cordyceps militaris.

The invention relates to a method for cultivating Cordyceps militaris, comprising:

Step 1: Take a predetermined weight of potato flakes and add water to boil and filter to obtain a potato filtrate. Then, add a predetermined weight of glucose and a predetermined weight of the agar, and then autoclave. After cooling, pour into a petri dish and separate and purify it. Cordyceps sinensis;

Step 2: taking a predetermined weight of potato flakes, adding water to boil and filtering to obtain a potato filtrate, and then adding a predetermined weight of glucose to be heated again until the glucose is completely melted to obtain a liquid medium; the liquid medium is autoclaved and then cooled for use; The Cordyceps militaris species is cut into the bacterial species, inoculated into the liquid medium, and shake cultured to obtain a liquid seed culture for cultivation;

Step 3: dissolving potassium dihydrogen phosphate, magnesium sulfate, ammonium citrate, glucose, peptone and vitamin B1 in the liquid medium to prepare a nutrient solution;

Step 4: pouring a predetermined proportion of white rice and the nutrient solution into a stainless steel container and performing high temperature sterilization, cooling and inoculating the culture liquid inoculum to obtain a solid culture material, and then sterilizing the solid culture material. Filling into one of the sterile plastic containers that have been sterilized;

Step 5: The plastic container is statically placed in a dark environment having a predetermined temperature and a predetermined humidity, so that the surface of the solid culture material grows hyphae, and after a predetermined time, the temperature of the dark environment is raised, and then Cultivate for a predetermined time;

Step 6: The plastic container is placed in a day and night to change the environment for a predetermined time, and the hyphae is changed by daytime and nighttime illumination and temperature difference, wherein the day and night changing environment has a temperature and illumination greater than that in the evening. Have temperature and illuminance;

Step 7: The plastic container is placed in a constant environment for a predetermined time to complete the cultivation of Cordyceps militaris, wherein the constant environment has stable temperature and humidity, and the light is irradiated to the hyphae.

Through the above steps, a simple and stable batch of Cordyceps militaris fruit body production and a cost-effective cultivation method of Cordyceps militaris are achieved.

Please refer to FIG. 1 , a method for cultivating Cordyceps militaris according to a preferred embodiment of the present invention, comprising the following steps:

Step 1: Purify the strain:

In this step, the potato is first peeled and cut into potato slices with a thickness of about 0.2 cm, 300 g of water is weighed, 1 liter of fresh water is added, and after boiling, it is filtered with gauze to obtain a potato filtrate, followed by 20 g of glucose and 15 g of ocean. After the dish is boiled, the volume is adjusted to 1000 ml, and autoclaved at 121 °C for 30 minutes. After cooling, it is poured into a Petri dish and separated and purified to obtain Cordyceps militaris.

Step 2: Production of inoculum:

A potato chip having a thickness of about 0.2 cm after peeling of 300 g of potato was weighed, and 1 liter of fresh water was added thereto. After heating to boiling, it was filtered with gauze to obtain a potato filtrate. 20 g of glucose was added to the potato filtrate and heated again until the glucose was completely melted to obtain a liquid medium. 1000 ml of liquid medium was taken, autoclaved at 121 ° C for 30 minutes, cooled and taken out for use. The Cordyceps militaris species was cut into 0.5 cm strains, inoculated into the liquid medium, and then the liquid medium was sent to an oscillating device, and cultured at 120 rpm and 21 ° C for 5 days, so that the liquid medium would A uniform pellet is formed to obtain a liquid inoculum for culture.

Step 3: Prepare nutrient solution:

The nutrient solution comprises 2 g of potassium dihydrogen phosphate, 1 g of magnesium sulfate, 1 g of ammonium citrate, 30 g of glucose, 3 g of peptone, 50 mg of vitamin B1, dissolved in 200 ml of the liquid medium, and diluted. Up to 1000 ml.

Step 4: Prepare solid culture material:

Pour 50 g of white rice into a closed stainless steel container, add 75 ml of the nutrient solution, sterilize at 121 ° C, 15 lb / cm ^{2 for} 30 minutes, cool and inoculate the culture liquid to inoculate, stir and homogenize it to obtain The solid culture material. The solid culture material is then aseptically filled into a sterilized sterile plastic container (as shown in Figure 2). In one embodiment, the plastic container may be polypropylene (PP), polyethylene (PE), high density polyethylene (HDPE) or polyethylene terephthalate (ET). production.

Step 5: Cultivate:

The mycelial growth stage needs to be protected from light. Therefore, the plastic container is placed in a dark environment at a temperature of 15-18 ° C and a humidity of 60%. After 5 days, the surface of the solid culture material grows hyphae and the temperature is raised to 20 At -23 ° C, culture for another 15 days to complete vegetative growth.

Step 6: Color change steps:

The plastic container is placed in a day and night changing environment, and the daylight and nighttime illumination and temperature difference are used to stimulate the hyphae to change color, and the temperature difference between day and night is controlled at about 10 °C, and the light is increased during the day, 450 Lux, temperature 21 ° C, and the light source is turned off at night. The temperature was set at 11 ° C, and the temperature difference between day and night was stimulated and the light was changed. After 7 days of culture, the mycelium was changed from white to golden yellow, and the primordium was gradually formed (as shown in Fig. 3).

Step 7: Complete the cultivation:

The plastic container is placed in a constant environment, the ventilation is ventilated, the temperature is maintained at 18~21 °C, no temperature difference stimulation is required, the relative humidity is 80%~855%, and the northern Cordyceps has phototaxis, so that the light source can shine all The fruiting body of the mycelium has completed the cultivation of Cordyceps militaris (as shown in Figure 4).

The method for cultivating Cordyceps militaris provided by the invention utilizes a closed stainless steel container as an intermediate conversion container for cultivation, so that the subsequent hyphal cultivation can be completed in the plastic container, thereby greatly simplifying the equipment required for cultivating Cordyceps militaris and reducing the equipment. The cost and the competitiveness of the industry are hereby submitted.

no

1 is a flow chart of a preferred embodiment of the present invention. 2 is a photograph of a strain cultured in a plastic container according to a preferred embodiment of the present invention. Fig. 3 is a photograph showing the color change of a strain in a plastic container according to a preferred embodiment of the present invention. Fig. 4 is a photograph showing the grass of the present invention in a preferred embodiment.

Claims (10)

A method for cultivating Cordyceps militaris comprises the following steps: Step 1: Take a predetermined weight of potato flakes, add water, boil and filter to obtain a potato filtrate, and then add a predetermined weight of glucose and a predetermined weight of the seaweed to be boiled and then autoclaved. After cooling, pour into a petri dish and separate and purify to obtain Cordyceps militaris; Step 2: Take a predetermined weight of potato flakes, add water, boil and filter to obtain a potato filtrate, then add a predetermined weight of glucose and heat again until glucose is completely melted. a liquid medium; the liquid medium is autoclaved and then cooled for use; the Cordyceps militaris species is cut into a bacterial mass, inoculated into the liquid medium, and shake cultured to obtain a liquid seed for cultivation; Step 3: Phosphoric acid Hydrogen potassium, magnesium sulfate, ammonium citrate, glucose, peptone and vitamin B1 are dissolved in the liquid medium to form a nutrient solution. Step 4: Pour a predetermined proportion of white rice and the nutrient solution into a stainless steel container and High temperature sterilization, cooling and inoculating to the culture liquid inoculum Obtaining a solid culture material, and then filling the solid culture material into one of the sterilized plastic containers in a sterile state; Step 5: statically placing the plastic container in a dark place having a predetermined temperature and a predetermined humidity Environment, the surface of the solid culture material grows hyphae, and after a predetermined time, the temperature of the dark environment is raised, and then cultured for a predetermined time; Step 6: the plastic container is placed in a day and night to change the environment for a predetermined time. Using daytime and nighttime illumination and temperature difference stimulation to promote the color change of the hyphae, wherein the temperature and illumination of the day and night changing environment during the day is greater than the temperature and illumination of the night; Step 7: statically placing the plastic container The constant environment is for a predetermined period of time to complete the cultivation of Cordyceps militaris, wherein the constant environment has a stable temperature and humidity, and the light is irradiated to the hyphae. The method for cultivating Cordyceps militaris according to claim 1, wherein in the first step, the potato sheet has a thickness of 0.2 cm and a weight of 300 gram; the added water is 1 liter; the added glucose is 20 gram, and the leaf is 15 g, dilute to 1000 ml after boiling; autoclave temperature is 121 ° C for 30 minutes. The method for cultivating Cordyceps militaris according to claim 1, wherein in the second step, the potato flakes have a thickness of 0.2 cm and a weight of 300 g; the added water is 1 liter; the added glucose is 20 g, and after boiling The volume was adjusted to 1000 ml; the autoclaving temperature was 121 ° C for 30 minutes; the shaking culture was carried out at 120 rpm and 21 ° C for 5 days. The method for cultivating Cordyceps militaris according to claim 1, wherein in the third step, the nutrient solution comprises 2 g of potassium dihydrogen phosphate, 1 g of magnesium sulfate, 1 g of ammonium citrate, and 30 g of glucose. The peptide was 3 g and the vitamin B1 was 50 mg. It was dissolved in 200 ml of the liquid medium and diluted to 1000 ml. The method for cultivating Cordyceps militaris according to claim 1, wherein in the fourth step, the weight of the white rice is 50 g, 75 ml of the nutrient solution is added, and sterilization is carried out at 121 ° C, 15 lb / cm ^{2 for} 30 minutes. The method for cultivating Cordyceps militaris according to claim 1, wherein the plastic container is made of polypropylene (polypropylene, PP), polyethylene (PE), high density polyethylene (HDPE) or polyethylene terephthalate. Any of the esters (Polyethylene terephthalate, ET). The method for cultivating Cordyceps militaris according to claim 1, wherein in the step 5, the temperature of the dark environment is 15-18 ° C, the humidity is 60%, the cultivating is 5 days, and then the temperature is raised to 20-23 ° C, and then cultured. 15 days. The method for cultivating Cordyceps militaris according to claim 1, wherein in the step 6, the diurnal temperature difference of the day and night changing environment is 10 °C. The method for cultivating Cordyceps militaris according to claim 1, wherein in the step 6, the daytime temperature of the day and night changing environment is 21 ° C, the illuminance is 450 Lux; and the light source is turned off at night, the temperature is 11 ° C. The method for cultivating Cordyceps militaris according to claim 1, wherein the constant environment temperature is 18 to 21 ° C, and the relative humidity is 80% to 85%.