CN108264390A - A kind of industrial planting method and its culture medium of Phellinus fructification - Google Patents

Abstract

The invention discloses a method for industrialized cultivation of Phellinus chinensis fruiting bodies and its culture medium, which comprises the steps of preparing Phellinus japonica seeds, preparing medium for industrialized production, inoculating, cultivating and managing, harvesting, and drying, wherein solid strains are expanded The medium includes corn flour, maltose, peptone, MgSO4, KH2P04, agar; the liquid strain expansion medium includes glucose, yeast powder, MgSO4, KH2PO4, triammonium citrate, mulberry branch water extract, vitamin B1; industrial production culture The base includes grain seeds, distilled water, MgSO4, KH2P04, and water extract of mulberry branches. The invention has low pollution rate, short cultivation period, stable product quality and low cost, and can realize industrialized and standardized production. The culture medium makes Phellinus mycelia grow fast and vigorously, and it takes a short time to obtain a large amount of Phellinus seeds. The raw materials are simple and easy to purchase, and the price is low, so that the cultivation time of Phellinus fruiting bodies is shortened, the biotransformation rate is high, and the polysaccharides, The content of flavonoids and three posts is relatively high.

Description

A kind of industrialized cultivation method of Phellinus chinensis fruiting body and culture medium thereof

Technical field:

The invention relates to a method for industrialized cultivation of the fruiting body of Phellinus phellinus and its culture medium, which belongs to the technical field of cultivation of edible and medicinal fungi, and relates to a method for cultivating the famous and precious medicinal fungus Phellinus phellidis. By adopting the method, the industrialization of Phellinus phellidis can be realized , Standardization, aseptic cultivation.

Background technique:

Phellinus igniarius, also known as Phellinus igniarius , Phellinus igniarius, Phellinus igniarius, Plum tree fungus, belongs to Basidiomycotina , Hymenomycetes , Polyporaceae , wood layer Puccinia pusilla is a rare medicinal fungus born on the trunks of broad-leaved trees such as poplars and willows. It grows in North China, Northwest China, Heilongjiang, Jilin, Taiwan, Guangdong, Sichuan, Yunnan, Tibet and other regions. Guang, is a precious Chinese herbal medicine. Due to the complexity and particularity of Phellinus Phellinus physiological ecology and the uncertainty of external environmental conditions, it takes many years to form Phellinus Phellinus fruiting bodies in nature, and due to over-harvesting, wild Phellinus Phellinus is very rare, and the price was once as high as per 5000~8000 RMB per kilogram.

Phellinus Phellinus is rich in flavonoids, polysaccharides, triterpenoids and other active ingredients, which have the functions of anti-oxidation, anti-tumor, anti-inflammation, enhancing immunity and repairing cell functions, and have significant hemostasis, blood circulation, and drinking , antidiarrheal and other effects. At the same time, Phellinus was confirmed to be non-toxic. Feeding experiments on mice showed no abnormal phenomena, and it had a detoxifying effect on liver alcoholism in mice.

At present, the artificial cultivation of Phellinus japonica has been realized, and it is cultivated in South Korea, China, and Japan. The method is to use basswood or mushroom bag cultivation. It takes more than three years to harvest the fruiting bodies. Due to the long cultivation period and environmental exposure, the pollution rate is high. The product quality is uneven and the price of finished products is high. The comprehensive factors of output, quality and price limit the further development and application of Phellinus.

Especially the cultivation period of the existing Phellinus radix fruiting bodies is too long, and it is impossible to realize industrialized and mechanized cultivation, and the industrialized cultivation of Phellinus radix is still blank. How to develop a Phellinus japonica cultivation technology that has a short cultivation period, can be standardized, industrialized, and mechanized, effectively solves the pollution problem in the cultivation process, maintains stable product quality, and greatly reduces production costs has become a key problem to be solved urgently.

Invention content:

The object of the present invention is to overcome above-mentioned deficiencies in the prior art and provide a kind of factory cultivation method of Phellinus radix fruiting body and culture medium thereof, the whole process of this cultivation method is cultivated in the bottle, the pollution rate is low, the cultivation period is short, and the product quality is high. Stable, low cost, can realize factory and standardized production. The used solid strain expansion medium Phellinus mycelia grows the fastest, the mycelium is darker in color, is not easy to degenerate into white, and the mycelium is the most vigorous and dense; the liquid strain expansion medium used can be used in a short time A large amount of Phellinus japonica seeds are obtained in the field; the raw material of the industrial production medium used is simple and easy to purchase, and the price is low, which shortens the cultivation time of the Phellinus japonica fruiting bodies, and has a high biotransformation rate, which can keep the content of polysaccharides, flavonoids, and three ingredients in it relatively low. high.

The purpose of the present invention can be achieved through the following measures:

A solid strain expansion medium for industrialized cultivation of Phellinus radix fruiting bodies, characterized in that the medium raw materials include: 10-50g/L corn flour, 10-50g/L maltose, 0.5-5g/L peptone, O.1-2.0g/L MgSO4, 1-6g/LKH2P04, 15-25g/L agar, balance water, natural pH; this medium is the most suitable solid strain expansion culture for the growth of Phellinus found through screening On this medium, Phellinus mycelium grows the fastest, mycelium is darker in color, not easy to degenerate into white, and the mycelium is the most vigorous and dense, which will lay the foundation for the cultivation of Phellinus japonica into fruiting bodies.

A liquid strain expansion medium for industrialized cultivation of Phellinus chinensis, characterized in that the medium raw materials include: 30-50g/L glucose, 5-15g/L yeast powder, 1-5g/L MgSO4, 1-5g/L KH2PO4, 1-5g/L triammonium citrate, 1-5g/L Mori twig water extract, 0.05-3g/L vitamin B1, balance water, natural pH; The most suitable liquid strain expansion medium for the growth of Phellinus found through screening can obtain a large number of Phellinus seeds in a short period of time, laying the foundation for reducing the production cycle and large-scale industrial production of Phellinus fruiting bodies.

An industrialized production medium for industrialized cultivation of Phellinus radix fruiting bodies is characterized in that the raw material of the medium comprises: in terms of weight percentage of the dark medium, it contains: 40-50% non-mildew grain seeds, 40-50% % distilled water, 0.1-3.0%MgSO4, 0.1-3.0%KH2P04, 1-10% Mori twig water extract, pH is natural, and the grain seed described is a kind of in rice, millet, sorghum, wheat, oat, corn; The raw materials of the factory production medium are easy to purchase and the price is low. On this medium, the traditional cultivation time of Phellinus radix fruiting bodies can be shortened to 6 months. At the same time, the biotransformation rate is high, and the polysaccharides, flavonoids, three Post composition content is higher.

A kind of factory cultivation method of Phellinus radix fruiting body is characterized in that it comprises the following specific steps:

(1) Strain: Phellinus fungus, stored on PDA slant medium;

(2) Seed preparation: under conditions suitable for Phellinus growth, cultivate Phellinus mycelia in liquid seed medium to logarithmic growth phase to obtain liquid Phellinus seeds; or cultivate Phellinus mycelium in solid seed medium , to obtain solid Phellinus seeds; its concrete steps are as follows:

Activation of preserved strains: the solid seed medium for the activation of strains is carrot agar solid medium, inoculated with Phellinus slant strains at 28°C, and cultured in dark for 8-12 days;

Expansion culture of solid strains: solid strain expansion medium uses 10-20g/L corn flour, 10-20g/L maltose, 0.5-1g/L peptone, O.1-0.6g/L MgSO4, 1 -4g/L KH2P04, 15-25g/L agar, remaining water, natural pH; after inoculating the activated Phellinus bacterium, culture at 25°C, and finally obtain solid Phellinus seeds of sufficient quantity and high quality for the production workshop ;

Expanded cultivation of liquid strains: the components used in liquid strain expansion medium are 30-50g/L glucose, 5-15g/L yeast powder, 1-5g/L MgSO4, 1-5g/L KH2PO4, 1-5g/L L triammonium citrate, 1-5g/L Mori twig water extract, 0.05-3g/L vitamin B1, the rest water, natural pH; after inoculating the activated Phellinus fungus, culture at 20-30°C , rotating speed 120-130rpm, finally obtain the liquid Phellinus Phellinus seeds of sufficient quantity for the production workshop;

(3) Preparation of medium for factory production: dark medium by weight percentage, it contains: 40-50% non-mildew grain seeds, 40-50% distilled water, 0.1-3.0% MgSO4, 0.1-3.0% KH2P04, 1- 10% mulberry branch water extract, mix the above medium components evenly according to the ratio, cover and seal, then sterilize at 121°C, 103.4kPa for 1h, the grain seeds are rice, millet, sorghum, wheat, One of oats and corn;

(4) Inoculation, cultivation management, and harvesting: in a sterile environment, inoculate the prepared liquid Phellinus seeds or solid Phellinus seeds on the sterilized industrial production medium, and culture at 24-28°C Cultivate in the dark in the room for 6 months, and harvest the Phellinus fruiting bodies; during the dark cultivation process, the medium humidity is 62%-68%, and the air humidity is 60%-70%. Oxygen supply, requiring no light in the room;

(5) Drying: The harvested Phellinus chinensis fruiting bodies are dried in a constant temperature drying oven at 50°C-60°C for 12 hours, so that the water content of the Phellinus huang fruiting bodies reaches below 1.3%.

Compared with the prior art, the present invention can produce the following positive effects: the present invention utilizes the grain matrix culture medium to cultivate Phellinus spp., can obtain Phellinus fruiting bodies in a short time, and can well solve the problem that there are few precious medicinal fungi. The problem of few sources of material. At the same time, the obtained Phellinus fruiting bodies have high active ingredients, which can be used in the development of health care products and produce certain economic benefits; the whole cultivation process is cultivated in bottles, with low pollution rate, short cultivation period, stable product quality and low cost, which can realize Factory and standardized production.

Description of drawings:

Fig. 1 is a situation diagram of bottle-planting Phellinus radix fruiting bodies on different grain mediums of the present invention.

The specific embodiment: the specific embodiment of the present invention is described in detail below:

Embodiment 1: Rice is the main raw material.

(1) Strains: Phellinus strains, purchased from Guangdong Microbial Strain Collection Center, and stored on PDA slant medium.

(2) Seed preparation: under conditions suitable for Phellinus growth, cultivate Phellinus mycelia in liquid seed medium to logarithmic growth phase to obtain liquid Phellinus seeds; or cultivate Phellinus mycelium in solid seed medium , to obtain solid Phellinus phellidis seeds, its specific steps are as follows:

Activation of preserved strains: the solid seed medium used for the activation of strains is carrot agar solid medium, and after inoculation of slant strains, the temperature is 28°C, and dark culture is carried out for 8 days.

Expansion culture of solid strains: the composition of solid strain expansion medium is 20g/L corn flour, 20g/L maltose, 1g/L peptone, 0.6g/L MgSO4, 1g/L KH2P04, 15g/L agar, and Measure the water, the pH is natural; after inoculating the activated Phellinus spp., cultivate at 25°C, and finally obtain solid Phellinus seeds of sufficient quantity and high quality for the production workshop.

Expansion culture of liquid strains: the composition of liquid strain expansion medium is 30g/L glucose, 5g/L yeast powder, 1g/L MgSO4, 1g/L KH2PO4, 2g/L triammonium citrate, 1g/L mulberry Branch water extract, 0.05g/L vitamin B1, balance water, PH natural. After the activated Phellinus strains are inoculated, the culture temperature is 20° C., and the rotation speed is 120 rpm, and finally enough liquid Phellinus seeds of high quality are obtained for the production workshop.

(3) Preparation of industrial production medium: by weight percentage of the medium, it contains: 50% non-mildew rice seeds, 45% distilled water, 2.0% MgSO4, 2.0% KH2P04, 1% mulberry branch water extract, the The above medium components were mixed evenly according to the ratios mentioned above, and after being covered and sealed, sterilized at 121° C. and 103.4 kPa for 1 hour.

(4) Inoculation, cultivation management, and harvesting: in a sterile environment, inoculate the prepared liquid or solid Phellinus Phellinus seeds on the sterilized industrial production medium, and place them in a 26°C cultivation room After 6 months of dark culture, harvest Phellinus fruiting bodies; during the dark culture process, the humidity of the culture medium is 62%, and the air humidity is 68%. Light;

(5) Drying: The harvested Phellinus fruiting body is dried in a constant temperature drying oven at 60°C for 12 hours, so that the water content of the Phellinus fruiting body reaches below 1.3%.

Example results: wet weight: 180.90g/jin (medium); dry weight: 52g/jin (medium).

Use rice as the raw material to cultivate Phellinus, and use a certain method to extract the active ingredients. The content is: polysaccharide content: 19.49mg/g; flavonoid content: 11.82mg/g; triterpene content: 0.43mg/g.

Embodiment 2: with millet main raw material

(1) Strains: Phellinus strains, purchased from Guangdong Microbial Strain Collection Center, and stored on PDA slant medium.

(2) Seed preparation: under conditions suitable for Phellinus growth, cultivate Phellinus mycelia in liquid seed medium to logarithmic growth phase to obtain liquid Phellinus seeds; or cultivate Phellinus mycelium in solid seed medium , to obtain solid Phellinus phellidis seeds, its specific steps are as follows:

Activation of preserved strains: the solid seed medium used for the activation of strains is carrot agar solid medium, and after inoculation of slant strains, the temperature is 28°C, and dark culture is carried out for 12 days.

Expansion culture of solid strains: the composition of solid strain expansion medium is 15g/L corn flour, 15g/L maltose, 0.8g/L peptone, 0.5g/L MgSO4, 3g/L KH2P04, 20g/L agar, The remaining water has a natural pH; after the activated Phellinus strains are inoculated, the culture temperature is 25° C., and finally solid Phellinus seeds of sufficient quantity and high quality are obtained for the production workshop.

Expansion culture of liquid strains: liquid strain expansion medium uses 40g/L glucose, 10g/L yeast powder, 3g/L MgSO4, 2g/L KH2PO4, 1g/L triammonium citrate, 2g/L mulberry Branch water extract, 0.1g/L vitamin B1, balance water, PH natural. After the activated Phellinus strains are inoculated, the culture temperature is 25° C., and the rotation speed is 125 rpm, and finally enough liquid Phellinus seeds of high quality are obtained for the production workshop.

(3) Preparation of industrial production medium: by weight percentage of the medium, it contains: 45% non-mildew millet seeds, 40% distilled water, 2.5% MgSO4, 2.5% KH2P04, 10% mulberry branch water extract, will Mix the above medium components evenly according to the ratio, cover and seal, and then sterilize at 121°C and 103.4kPa for 1 hour;

(4) Inoculation, cultivation management, and harvesting: in a sterile environment, inoculate the prepared liquid Phellinus seeds or solid Phellinus seeds on the sterilized industrial production medium, and place them in a 24°C cultivation room After 6 months of dark culture, harvest Phellinus fruiting bodies; during the dark culture process, the humidity of the culture medium is 65%, and the air humidity is 60%. Light;

(5) Drying: The harvested Phellinus fruiting bodies were dried in a constant temperature drying oven at 50°C for 12 hours, so that the water content of the Phellinus fruiting bodies reached below 1.3%.

Example results: wet weight: 103.4g/jin (medium); dry weight: 27.1g/jin (medium).

Cultivate Phellinus using millet as the raw material, and use a certain method to extract the active ingredients. The content is: polysaccharide content: 10.77mg/g; flavonoid content: 3.9mg/g; triterpene content: 0.7mg/g.

Embodiment 3: with oat main raw material

(1) Strains: Phellinus strains, purchased from Guangdong Microbial Strain Collection Center, and stored on PDA slant medium.

(2) Seed preparation: under conditions suitable for Phellinus growth, cultivate Phellinus mycelia in liquid seed medium to logarithmic growth phase to obtain liquid Phellinus seeds; or cultivate Phellinus mycelium in solid seed medium , to obtain solid Phellinus phellidis seeds, its specific steps are as follows:

Activation of preserved strains: the solid seed medium used for the activation of strains is carrot agar solid medium, and after inoculation of slant strains, the temperature is 28°C, and dark culture is carried out for 10 days.

Expansion culture of solid strains: the composition of solid strain expansion medium is 10g/L corn flour, 10g/L maltose, 0.5g/L peptone, 0.1g/L MgSO4, 4g/L KH2P04, 25g/L agar, The remaining water has a natural pH; after the activated Phellinus strains are inoculated, the culture temperature is 25° C., and finally solid Phellinus seeds of sufficient quantity and high quality are obtained for the production workshop.

Expansion culture of liquid strains: The composition of liquid strain expansion medium is 50g/L glucose, 15g/L yeast powder, 5g/L MgSO4, 5g/L KH2PO4, 5g/L triammonium citrate, 5g/L mulberry Branch water extract, 3g/L vitamin B1, balance water, PH natural. After the activated Phellinus strains are inoculated, the culturing temperature is 30° C. and the rotation speed is 130 rpm, and finally enough liquid Phellinus seeds of high quality are obtained for the production workshop.

(3) Preparation of industrial production medium: by weight percentage of the medium, it contains: 40% non-moulded oat seeds, 50% distilled water, 3.0% MgSO4, 3.0% KH2P04, 4% mulberry branch water extract, the The above medium components were mixed evenly according to the ratios mentioned above, and after being covered and sealed, sterilized at 121° C. and 103.4 kPa for 1 hour.

(4) Inoculation, cultivation management, and harvesting: in a sterile environment, inoculate the prepared liquid Phellinus seeds or solid Phellinus seeds on the sterilized industrial production medium, and place them in a 28°C cultivation room After 6 months of dark culture, harvest Phellinus fruiting bodies; during the dark culture process, the humidity of the culture medium is 62%, and the air humidity is 65%. Light;

(5) Drying: The harvested Phellinus fruiting bodies were dried in a constant temperature drying oven at 55°C for 12 hours, so that the water content of the Phellinus fruiting bodies reached below 1.3%.

Example results: wet weight: 141.9g/jin (medium); dry weight: 33.85g/jin (medium).

Use oats as raw materials to cultivate Phellinus japonica, and use a certain method to extract the active ingredients. The content is: polysaccharide content: 8.85mg/g; flavonoid content: 2.29mg/g; triterpene content: 0.45mg/g.

Embodiment 4: with wheat bran main raw material

(1) Strains: Phellinus strains, purchased from Guangdong Microbial Strain Collection Center, and stored on PDA slant medium.

(2) Seed preparation: under conditions suitable for Phellinus growth, cultivate Phellinus mycelia in liquid seed medium to logarithmic growth phase to obtain liquid Phellinus seeds; or cultivate Phellinus mycelium in solid seed medium , to obtain solid Phellinus phellidis seeds, its specific steps are as follows:

Activation of preserved strains: the solid seed medium used for the activation of strains is carrot agar solid medium, and after inoculation of slant strains, the temperature is 28°C, and dark culture is carried out for 10 days.

Expansion culture of solid strains: the composition of solid strain expansion medium is 20g/L corn flour, 10g/L maltose, 0.5g/L peptone, 0.6g/L MgSO4, 3g/L KH2P04, 15g/L agar, The remaining water has a natural pH; after the activated Phellinus strains are inoculated, the culture temperature is 25° C., and finally solid Phellinus seeds of sufficient quantity and high quality are obtained for the production workshop.

Expansion culture of liquid strains: The composition of liquid strain expansion medium is 40g/L glucose, 15g/L yeast powder, 3g/L MgSO4, 3g/L KH2PO4, 3g/L triammonium citrate, 4g/L mulberry Branch water extract, 2g/L vitamin B1, balance water, PH natural. After the activated Phellinus strains are inoculated, the culture temperature is 25° C., and the rotation speed is 125 rpm, and finally enough liquid Phellinus seeds of high quality are obtained for the production workshop.

(3) Preparation of medium for factory production: In terms of weight percentage of medium, it contains: 45% non-moulded wheat bran, 45% distilled water, 0.1% MgSO4, 0.1% KH2P04, 9.8% water extract of Mori twig The above medium components were mixed evenly according to the ratios mentioned above, and after being covered and sealed, sterilized at 121° C. and 103.4 kPa for 1 hour.

(4) Inoculation, cultivation management, and harvesting: in a sterile environment, inoculate the prepared liquid or solid Phellinus Phellinus seeds on the sterilized industrial production medium, and place them in a 27°C cultivation room After 6 months of dark culture, harvest Phellinus fruiting bodies; during the dark culture process, the humidity of the culture medium is 68%, and the air humidity is 70%. Light;

(5) Drying: The harvested Phellinus fruiting body is dried in a constant temperature drying oven at 60°C for 12 hours, so that the water content of the Phellinus fruiting body reaches below 1.3%.

Example results: wet weight: 96.5g/catties (medium), dry weight 22.6g/catties (medium).

Using wheat bran as the culture material to cultivate Phellinus japonica, using certain methods to extract the active ingredients, the content is: polysaccharide content: 3.68mg/g; flavonoid content: 0.39mg/g; triterpene content: 0.47mg/g.

It should be understood that the parts not described in detail in this specification belong to the prior art. The above description of the preferred embodiments is more detailed, but it should not be considered as limiting the protection scope of the patent of the present invention.

Claims (4)

1. A solid strain expansion medium for the industrialized cultivation of Phellinus fruiting bodies, characterized in that the medium raw materials include: 10-50g/L corn flour, 10-50g/L maltose, 0.5-5g/L Peptone, O.1-2.0g/L MgSO4, 1-6g/LKH2P04, 15-25g/L agar, balance water, natural pH. 2. A liquid strain expansion medium for the industrialized cultivation of Phellinus fruiting bodies, characterized in that the medium raw materials include: 30-50g/L glucose, 5-15g/L yeast powder, 1-5g/L MgSO4, 1-5g/L KH2PO4, 1-5g/L triammonium citrate, 1-5g/L Mori twig water extract, 0.05-3g/L vitamin B1, balance water, natural pH. 3. An industrial production medium for the industrialized cultivation of Phellinus radix fruiting bodies, characterized in that the medium raw materials include: dark medium by weight percentage, 40-50% non-mildew grain seeds, 40-50% Distilled water, 0.1-3.0% MgSO4, 0.1-3.0% KH2P04, 1-10% water extract of Mori twigs, natural pH, and the grain seeds are one of rice, millet, sorghum, wheat, oats, and corn. 4. a factory cultivation method of Phellinus radix fruiting body, is characterized in that it comprises following concrete steps: (1) Strain: Phellinus fungus, stored on PDA slant medium; (2) Seed preparation: under conditions suitable for Phellinus growth, cultivate Phellinus mycelia in liquid seed medium to logarithmic growth phase to obtain liquid Phellinus seeds; or cultivate Phellinus mycelium in solid seed medium , to obtain solid Phellinus seeds; its concrete steps are as follows: Activation of preserved strains: the solid seed medium for the activation of strains is carrot agar solid medium, inoculated with Phellinus slant strains at 28°C, and cultured in dark for 8-12 days; Expansion culture of solid strains: solid strain expansion medium uses 10-20g/L corn flour, 10-20g/L maltose, 0.5-1g/L peptone, O.1-0.6g/L MgSO4, 1 -4g/L KH2P04, 15-25g/L agar, remaining water, natural pH; after inoculating the activated Phellinus bacterium, culture at 25°C, and finally obtain solid Phellinus seeds of sufficient quantity and high quality for the production workshop ; Expanded cultivation of liquid strains: the components used in liquid strain expansion medium are 30-50g/L glucose, 5-15g/L yeast powder, 1-5g/L MgSO4, 1-5g/L KH2PO4, 1-5g/L L triammonium citrate, 1-5g/L Mori twig water extract, 0.05-3g/L vitamin B1, the rest water, natural pH; after inoculating the activated Phellinus fungus, culture at 20-30°C , rotating speed 120-130rpm, finally obtain the liquid Phellinus Phellinus seeds of sufficient quantity for the production workshop; (3) Preparation of medium for factory production: dark medium by weight percentage, it contains: 40-50% non-mildew grain seeds, 40-50% distilled water, 0.1-3.0% MgSO4, 0.1-3.0% KH2P04, 1- 10% mulberry branch water extract, mix the above medium components evenly according to the ratio, cover and seal, then sterilize at 121°C, 103.4kPa for 1h, the grain seeds are rice, millet, sorghum, wheat, One of oats and corn; (4) Inoculation, cultivation management, and harvesting: in a sterile environment, inoculate the prepared liquid Phellinus seeds or solid Phellinus seeds on the sterilized industrial production medium, and culture at 24-28°C Cultivate in the dark in the room for 6 months, and harvest the Phellinus fruiting bodies; during the dark cultivation process, the medium humidity is 62%-68%, and the air humidity is 60%-70%. Oxygen supply, requiring no light in the room; (5) Drying: The harvested Phellinus chinensis fruiting bodies are dried in a constant temperature drying oven at 50°C-60°C for 12 hours, so that the water content of the Phellinus huang fruiting bodies reaches below 1.3%.