CN109601254B - Antrodia camphorata culture method capable of completely separating culture product from matrix - Google Patents
Antrodia camphorata culture method capable of completely separating culture product from matrix Download PDFInfo
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Abstract
The invention provides an antrodia camphorata cultivation method for completely separating a cultivation product from a matrix, which selects grain and antrodia camphorata sawdust to be mixed as the cultivation matrix, different grains are uniformly mixed and then placed on the lower layer, a small amount of antrodia camphorata sawdust is laid on the lower layer, a cylindrical groove is drilled in the middle of the cultivation matrix during inoculation, and a strain block is inoculated into the cylindrical groove. After the culture is finished, a compact mycelium layer with a certain thickness is formed on the surface of the culture substrate, the mycelium layer can be easily peeled off, a mycelium similar to a stipe is formed in the cylindrical groove, the upper half part of the mycelium is sampled, and the rest mycelium can continue to grow. By using the method, the problem that the culture product is difficult to be completely separated from the matrix by using a solid culture method is solved, wood chips mixed in the culture product can be removed, the yield and the content of the triterpenes can be improved, reference help is provided for the practice of the solid state fermentation culture of the antrodia camphorata, a foundation is provided for the subsequent large-scale fermentation culture, and the method is low in cost and simple and easy to operate.
Description
Technical Field
The invention relates to a culture method of antrodia camphorata, in particular to a culture method of antrodia camphorata with culture products and matrix completely separated.
Background
Antrodia camphorata (Antrodia camphorata)Antrodia camphorata) Belongs to the family of polyporaceae and the genus of Botrytis, and is a rare medicinal fungus specific to Taiwan. A notice of an implementation scheme (2016) for promoting the protection and development of traditional Chinese medicinal materials in Guangdong province [ 2016 ] 61 ] is issued in an office of civil administration department in Guangdong province in 2016, and Antrodia camphorata is listed as one of varieties for enhancing the resource protection of traditional Chinese medicinal materials, building an endangered scarce traditional Chinese medicinal material planting and breeding base and developing 20 characteristic traditional Chinese medicinal material resource evaluation and breeding technical research for key protection and development. The mycelium and fruiting body of Antrodia Camphorata are rich in physiologically active substances, including triterpenes, polysaccharides, maleic acid and succinic acid derivatives. Modern researchers find that antrodia camphorata not only has a protective effect on chemically-caused liver injury, but also has the effects of resisting cancer, resisting oxidation,Anti-inflammatory and immunoregulatory effects.
The antrodia camphorata has obvious efficacy, the market demand is greatly increased, the wild antrodia camphorata grows slowly, the yield is extremely low, and the antrodia camphorata host antrodia camphorata is a national conservation tree species, so that the antrodia camphorata is cultured by adopting an artificial culture mode at present. The artificial culture method mainly comprises a basswood culture method, a solid culture method and a liquid culture method. The basswood culture method has long growth period and difficult quality control, but the basswood culture method can obtain the antrodia cinnamomea fruiting body with similar components to the wild antrodia cinnamomea by culture; the liquid culture method has short culture period and high yield, but the culture product has lower content of effective components and can not be cultured to obtain the antrodia cinnamomea fruiting body; the solid culture method has moderate growth cycle and controllable quality, and when the culture reaches a certain time, the antrodia cinnamomea fruiting bodies with similar components to the wild antrodia cinnamomea can be obtained, but the culture products are difficult to separate from the matrix.
Disclosure of Invention
The invention aims to provide the antrodia camphorata culture method which is low in cost, simple in process, good in product quality and capable of completely separating a culture product from a matrix, aiming at the existing technical current situation.
In order to achieve the purpose, the invention adopts the following technical scheme:
the invention relates to an antrodia camphorata culture method for completely separating a culture product from a matrix, which mainly comprises the following steps:
(1) activating and inoculating raw materials: performing activated culture on Antrodia Camphorata strain at 4 deg.C on PDA plate culture medium for 15-30d, and perforating near the edge of mycelium layer with a perforator diameter of 0.8cm to obtain mycelium block for inoculation;
(2) preparing a solid fermentation medium: weighing a certain amount of cereal raw materials and cinnamomum kanehirae swamp wood chips, placing the cereal raw materials and the cinnamomum kanehirae swamp wood chips in a culture bottle, uniformly mixing the cereal raw materials, placing the cereal raw materials in the lower layer of the culture bottle, and flatly paving the cinnamomum kanehirae swamp wood chips on the lower layer of the culture bottle; mixing glucose and KH2PO4、MgSO4·7H2Dissolving O in distilled water to form solution containing glucose and inorganic salts, pouring the solution into culture flask, soaking solid material to water content of 50-70% for 2 hr, and placing into autoclaveSterilizing at 121 deg.C for 30 min;
(3) and (3) punching and inoculating: drilling a cylindrical groove with the depth of about 2cm at the center of the solid culture medium by using a glass rod, inoculating one surface of a raw material hypha block containing hypha in the cylindrical groove hole by adhering to a culture medium, and placing in a constant-temperature incubator for dark culture at 28 ℃;
(4) sampling by an aseptic super-clean bench: after culturing for a certain time, a compact mycelium layer which is 3-5cm thick and can be easily peeled off is formed on the surface of the substrate, sampling is carried out in a sterile super clean bench, and the rest mycelium in the culture medium can continue to grow.
In the scheme, the processing method of the cinnamomum kanehirae wood chips comprises the steps of placing the cinnamomum kanehirae wood chips outdoors for insolation, then sterilizing the cinnamomum kanehirae wood chips by using an autoclave, wherein the sterilization temperature is 121 ℃ and the sterilization time is 2 hours, and after sterilization, placing the cinnamomum kanehirae wood chips into an oven to be dried at 80 ℃.
Furthermore, the grain raw materials comprise 50g of rice and 2.5g of bran, and the dry materials are uniformly mixed and then are put into a culture bottle, wherein the volume of the culture bottle is 450 mL.
Further, the Cinnamomum kanehirae Hayata sawdust is prepared by pulverizing Cinnamomum kanehirae Hayata wood into 0.5cm sheet sawdust, weighing dried Cinnamomum kanehirae Hayata sawdust 0.5-7g, and spreading on the surface of grain matrix.
Further, in the solution containing glucose and inorganic salt, 5g of glucose is contained, KH2PO4Is 0.5g, MgSO4·7H2O was 0.5 g.
The invention has the beneficial effects that: the culture medium is prepared by mixing grains and the cinnamomum kanehirae wood chips, the formula not only provides necessary nutrient substances for the growth of the cinnamomum kanehirae, but also adds the cinnamomum kanehirae wood chips necessary for the wild growth of the cinnamomum kanehirae, so that the components of the culture medium are more comprehensive. And then selecting a culture medium with the water content of 70%, wherein the growth of the antrodia camphorata mycelium tends to accumulate on the surface of the substrate gradually along with the increase of the water content of the culture medium, the mycelium does not grow into the culture medium any more, in order to make the mycelium fully utilize the nutrition in the culture medium, a cylindrical groove is formed by punching in the middle of the culture medium during inoculation, and the strain block is inoculated into the cylindrical groove. After the culture is finished, a compact mycelium layer with a certain thickness is formed on the surface of the culture substrate, the mycelium layer can be easily peeled from the substrate, meanwhile, the mycelium is combined in the cylindrical groove to form a mycelium similar to a stipe, the upper half part of the mycelium is sampled, and the rest mycelium can continue to grow in the substrate. The method can harvest a large amount of culture product on the surface of the substrate, and the culture product is freeze-dried after sampling, and the obtained fungus cake is mixed with a small amount of wood dust and can be easily removed.
Description of the drawings:
FIG. 1 shows the growth of Antrodia camphorata on a basic culture medium with different water contents (50 d), wherein the water contents are 50%, 60% and 70% from left to right;
FIG. 2 shows the growth of Antrodia camphorata (60d) on a culture medium added with different Antrodia camphorata sawdust, wherein the added amount of the Antrodia camphorata sawdust of 1-5g is 0.5 g/bottle, 1 g/bottle, 3 g/bottle, 5 g/bottle and 7 g/bottle respectively;
FIG. 3 shows Antrodia camphorata growing in a culture flask, with grain culture medium on the left and grain plus wood chips culture medium on the right;
FIG. 4 shows the lyophilized Antrodia camphorata cake.
The specific implementation mode is as follows:
example 1:
a culture product and substrate completely separated antrodia camphorata culture method mainly comprises the following steps:
(1) activating and inoculating raw materials: performing activated culture on Antrodia camphorata strain stored at 4 ℃ in a PDA plate culture medium for 15 days, and punching at a position 0.4cm away from the outer edge of a mycelium layer at the center of a puncher to obtain a mycelium block for inoculation;
(2) preparing a solid fermentation medium: weighing a certain amount of cereal raw materials and cinnamomum kanehirae wood chips, placing the cereal raw materials comprising 50g of rice and 2.5g of bran into a culture bottle, uniformly mixing the dry materials, putting the mixture into the culture bottle, wherein the volume of the culture bottle is 450mL, uniformly mixing the cereal raw materials, placing the mixture at the lower layer of the culture bottle, and flatly paving the cinnamomum kanehirae wood chips on the culture bottle; adding glucose 5g and KH2PO40.5g，MgSO4·7H2Dissolving O0.5 g in distilled water to obtain solution containing glucose and inorganic salts, pouring the solution into culture flask, soaking solid material to obtain culture medium with final water content of 60% for 2 hr, and sterilizing in autoclaveSterilizing at 121 deg.C for 30 min;
the processing method of the camphor tree wood dust comprises the steps of placing the camphor tree wood dust outdoors for insolation, then sterilizing the camphor tree wood dust in an autoclave for 2 hours at the temperature of 121 ℃, and then drying the camphor tree wood dust in an oven at the temperature of 80 ℃. The Cinnamomum kanehirai Hayata sawdust is prepared by pulverizing Cinnamomum kanehirai Hayata wood into 0.5cm pieces, weighing dried Cinnamomum kanehirai Hayata sawdust 3g, and spreading on the surface of grain matrix.
(3) And (3) punching and inoculating: drilling a cylindrical groove with the depth of about 2cm at the center of the solid culture medium by using a glass rod, inoculating one surface of a raw material hypha block containing hypha in the cylindrical groove hole by adhering to a culture medium, and placing in a constant-temperature incubator for dark culture at 28 ℃;
(4) sampling by an aseptic super-clean bench: after culturing for a certain time, a compact mycelium layer which is 3-5cm thick and can be easily peeled off is formed on the surface of the substrate, sampling is carried out in a sterile super clean bench, and the rest mycelium in the culture medium can continue to grow.
Example 2:
a culture product and substrate completely separated antrodia camphorata culture method mainly comprises the following steps:
(1) activating and inoculating raw materials: performing activated culture on Antrodia camphorata strain stored at 4 ℃ on a PDA plate culture medium for 20 days, and punching a hole at a position 0.4cm away from the outer edge of a mycelium layer at the center of the puncher to obtain a mycelium block for inoculation;
(2) preparing a solid fermentation medium: weighing a certain amount of cereal raw materials and cinnamomum kanehirae wood chips, placing the cereal raw materials comprising 50g of rice and 2.5g of bran into a culture bottle, uniformly mixing the dry materials, putting the mixture into the culture bottle, wherein the volume of the culture bottle is 450mL, uniformly mixing the cereal raw materials, placing the mixture at the lower layer of the culture bottle, and flatly paving the cinnamomum kanehirae wood chips on the culture bottle; adding glucose 5g and KH2PO40.5g，MgSO4·7H2Dissolving 0.5g of O in a certain amount of distilled water to form a solution containing glucose and inorganic salts, pouring the solution into a culture bottle, soaking the solid materials to ensure that the final water content of the culture medium is 70%, wherein the soaking time is 2h, and then placing the culture medium into an autoclave for sterilization at the sterilization temperature of 121 ℃ for 30 min;
the processing method of the camphor tree wood dust comprises the steps of placing the camphor tree wood dust outdoors for insolation, then sterilizing the camphor tree wood dust in an autoclave for 2 hours at the temperature of 121 ℃, and then drying the camphor tree wood dust in an oven at the temperature of 80 ℃. The Cinnamomum kanehirai Hayata sawdust is prepared by pulverizing Cinnamomum kanehirai Hayata wood into 0.5cm pieces, weighing dried Cinnamomum kanehirai Hayata sawdust 1g, and spreading on the surface of grain matrix.
(3) And (3) punching and inoculating: drilling a cylindrical groove with the depth of about 2cm at the center of the solid culture medium by using a glass rod, inoculating one surface of a raw material hypha block containing hypha in the cylindrical groove hole by adhering to a culture medium, and placing in a constant-temperature incubator for dark culture at 28 ℃;
(4) sampling by an aseptic super-clean bench: after culturing for a certain time, a compact mycelium layer which is 3-5cm thick and can be easily peeled off is formed on the surface of the substrate, sampling is carried out in a sterile super clean bench, and the rest mycelium in the culture medium can continue to grow.
Example 3:
a culture product and substrate completely separated antrodia camphorata culture method mainly comprises the following steps:
(1) activating and inoculating raw materials: performing activated culture on Antrodia camphorata strain stored at 4 ℃ on a PDA plate culture medium for 25d, and punching a hole at a position 1.2cm away from the outer edge of a mycelium layer in the center of a puncher to obtain a mycelium block for inoculation;
(2) preparing a solid fermentation medium: weighing a certain amount of cereal raw materials and cinnamomum kanehirae wood chips, placing the cereal raw materials comprising 50g of rice and 2.5g of bran into a culture bottle, uniformly mixing the dry materials, putting the mixture into the culture bottle, wherein the volume of the culture bottle is 450mL, uniformly mixing the cereal raw materials, placing the mixture at the lower layer of the culture bottle, and flatly paving the cinnamomum kanehirae wood chips on the culture bottle; adding glucose 5g and KH2PO40.5g，MgSO4·7H2Dissolving 0.5g of O in a certain amount of distilled water to form a solution containing glucose and inorganic salts, pouring the solution into a culture bottle, soaking the solid materials to ensure that the final water content of the culture medium is 50%, wherein the soaking time is 2h, and then placing the culture medium into an autoclave for sterilization at the sterilization temperature of 121 ℃ for 30 min;
the processing method of the camphor tree wood dust comprises the steps of placing the camphor tree wood dust outdoors for insolation, then sterilizing the camphor tree wood dust in an autoclave for 2 hours at the temperature of 121 ℃, and then drying the camphor tree wood dust in an oven at the temperature of 80 ℃. The Cinnamomum kanehirai Hayata sawdust is prepared by pulverizing Cinnamomum kanehirai Hayata wood into 0.5cm pieces, weighing dried Cinnamomum kanehirai Hayata sawdust 5g, and spreading on the surface of grain matrix.
(3) And (3) punching and inoculating: drilling a cylindrical groove with the depth of about 2cm at the center of the solid culture medium by using a glass rod, inoculating one surface of a raw material hypha block containing hypha in the cylindrical groove hole by adhering to a culture medium, and placing in a constant-temperature incubator for dark culture at 28 ℃;
(4) sampling by an aseptic super-clean bench: after culturing for a certain time, a compact mycelium layer which is 3-5cm thick and can be easily peeled off is formed on the surface of the substrate, sampling is carried out in a sterile super clean bench, and the rest mycelium in the culture medium can continue to grow.
Example 4:
a culture product and substrate completely separated antrodia camphorata culture method mainly comprises the following steps:
(1) activating and inoculating raw materials: performing activated culture on Antrodia camphorata strain stored at 4 ℃ on a PDA plate culture medium for 30 days, and punching a hole at a position 1.2cm away from the outer edge of a mycelium layer in the center of a puncher to obtain a mycelium block for inoculation;
(2) preparing a solid fermentation medium: weighing a certain amount of cereal raw materials and cinnamomum kanehirae wood chips, placing the cereal raw materials comprising 50g of rice and 2.5g of bran into a culture bottle, uniformly mixing the dry materials, putting the mixture into the culture bottle, wherein the volume of the culture bottle is 450mL, uniformly mixing the cereal raw materials, placing the mixture at the lower layer of the culture bottle, and flatly paving the cinnamomum kanehirae wood chips on the culture bottle; adding glucose 5g and KH2PO40.5g，MgSO4·7H2Dissolving 0.5g of O in a certain amount of distilled water to form a solution containing glucose and inorganic salts, pouring the solution into a culture bottle, soaking the solid materials to ensure that the final water content of the culture medium is 60%, wherein the soaking time is 2h, and then placing the culture medium into an autoclave for sterilization at the sterilization temperature of 121 ℃ for 30 min;
the processing method of the camphor tree wood dust comprises the steps of placing the camphor tree wood dust outdoors for insolation, then sterilizing the camphor tree wood dust in an autoclave for 2 hours at the temperature of 121 ℃, and then drying the camphor tree wood dust in an oven at the temperature of 80 ℃. The Cinnamomum kanehirae Hayata sawdust is prepared by pulverizing Cinnamomum kanehirae Hayata wood into 0.5cm pieces, weighing dried Cinnamomum kanehirae Hayata sawdust 0.5g, and spreading on the surface of grain matrix.
(3) And (3) punching and inoculating: drilling a cylindrical groove with the depth of about 2cm at the center of the solid culture medium by using a glass rod, inoculating one surface of a raw material hypha block containing hypha in the cylindrical groove hole by adhering to a culture medium, and placing in a constant-temperature incubator for dark culture at 28 ℃;
(4) sampling by an aseptic super-clean bench: after culturing for a certain time, a compact mycelium layer which is 3-5cm thick and can be easily peeled off is formed on the surface of the substrate, sampling is carried out in a sterile super clean bench, and the rest mycelium in the culture medium can continue to grow.
Example 5:
a culture product and substrate completely separated antrodia camphorata culture method mainly comprises the following steps:
(1) activating and inoculating raw materials: performing activated culture on Antrodia camphorata strain stored at 4 ℃ on a PDA plate culture medium for 20 days, and punching a hole at a position 0.4cm away from the outer edge of a mycelium layer at the center of the puncher to obtain a mycelium block for inoculation;
(2) preparing a solid fermentation medium: weighing a certain amount of cereal raw materials and cinnamomum kanehirae wood chips, placing the cereal raw materials comprising 50g of rice and 2.5g of bran into a culture bottle, uniformly mixing the dry materials, putting the mixture into the culture bottle, wherein the volume of the culture bottle is 450mL, uniformly mixing the cereal raw materials, placing the mixture at the lower layer of the culture bottle, and flatly paving the cinnamomum kanehirae wood chips on the culture bottle; adding glucose 5g and KH2PO40.5g，MgSO4·7H2Dissolving 0.5g of O in a certain amount of distilled water to form a solution containing glucose and inorganic salts, pouring the solution into a culture bottle, soaking the solid materials to ensure that the final water content of the culture medium is 70%, wherein the soaking time is 2h, and then placing the culture medium into an autoclave for sterilization at the sterilization temperature of 121 ℃ for 30 min;
the processing method of the camphor tree wood dust comprises the steps of placing the camphor tree wood dust outdoors for insolation, then sterilizing the camphor tree wood dust in an autoclave for 2 hours at the temperature of 121 ℃, and then drying the camphor tree wood dust in an oven at the temperature of 80 ℃. The Cinnamomum kanehirai Hayata sawdust is prepared by pulverizing Cinnamomum kanehirai Hayata wood into 0.5cm pieces, weighing dried Cinnamomum kanehirai Hayata sawdust 7g, and spreading on the surface of grain matrix.
(3) And (3) punching and inoculating: drilling a cylindrical groove with the depth of about 2cm at the center of the solid culture medium by using a glass rod, inoculating one surface of a raw material hypha block containing hypha in the cylindrical groove hole by adhering to a culture medium, and placing in a constant-temperature incubator for dark culture at 28 ℃;
(4) sampling by an aseptic super-clean bench: after culturing for a certain time, a compact mycelium layer which is 3-5cm thick and can be easily peeled off is formed on the surface of the substrate, sampling is carried out in a sterile super clean bench, and the rest mycelium in the culture medium can continue to grow.
Example 6:
a culture product and substrate completely separated antrodia camphorata culture method mainly comprises the following steps:
(1) activating and inoculating raw materials: performing activated culture on Antrodia camphorata strain stored at 4 ℃ on a PDA plate culture medium for 20 days, and punching a hole at a position 1.2cm away from the outer edge of a mycelium layer in the center of a puncher to obtain a mycelium block for inoculation;
(2) preparing a solid fermentation medium: weighing a certain amount of cereal raw materials and cinnamomum kanehirae wood chips, placing the cereal raw materials comprising 50g of rice and 2.5g of bran into a culture bottle, uniformly mixing the dry materials, putting the mixture into the culture bottle, wherein the volume of the culture bottle is 450mL, uniformly mixing the cereal raw materials, placing the mixture at the lower layer of the culture bottle, and flatly paving the cinnamomum kanehirae wood chips on the culture bottle; adding glucose 5g and KH2PO40.5g，MgSO4·7H2Dissolving 0.5g of O in a certain amount of distilled water to form a solution containing glucose and inorganic salts, pouring the solution into a culture bottle, soaking the solid materials to ensure that the final water content of the culture medium is 60%, wherein the soaking time is 2h, and then placing the culture medium into an autoclave for sterilization at the sterilization temperature of 121 ℃ for 30 min;
the processing method of the camphor tree wood dust comprises the steps of placing the camphor tree wood dust outdoors for insolation, then sterilizing the camphor tree wood dust in an autoclave for 2 hours at the temperature of 121 ℃, and then drying the camphor tree wood dust in an oven at the temperature of 80 ℃. The Cinnamomum kanehirai Hayata sawdust is prepared by pulverizing Cinnamomum kanehirai Hayata wood into 0.5cm pieces, weighing dried Cinnamomum kanehirai Hayata sawdust 1g, and spreading on the surface of grain matrix.
(3) And (3) punching and inoculating: drilling a cylindrical groove with the depth of about 2cm at the center of the solid culture medium by using a glass rod, inoculating one surface of a raw material hypha block containing hypha in the cylindrical groove hole by adhering to a culture medium, and placing in a constant-temperature incubator for dark culture at 28 ℃;
(4) sampling by an aseptic super-clean bench: after culturing for a certain time, a compact mycelium layer which is 3-5cm thick and can be easily peeled off is formed on the surface of the substrate, sampling is carried out in a sterile super clean bench, and the rest mycelium in the culture medium can continue to grow.
Comparing the growth differences of the Antrodia camphorata mycelium at different activation times (15 d, 20d, 25d, 30 d) and at a distance of 0.4cm and 1.2cm from the edge of the mycelium layer at the center of the perforation, the following table 1 is provided:
TABLE 1 different activation methods inoculation feedstock growth 30d growth diameter (cm)
As can be seen from Table 1, the mycelia of Antrodia camphorata were grown optimally by performing the activated culture for 20 days, and punching a hole in the center of the punch at a distance of 0.4cm from the edge of the mycelium layer (i.e., near the outer edge of the mycelium layer) to obtain an inoculum cake.
Examining the influence of different concentrations of the wood chips of the antrodia camphorata (0.5 g/bottle, 1 g/bottle, 3 g/bottle, 5 g/bottle and 7 g/bottle) on the growth condition of the antrodia camphorata by the basic culture media (50%, 60% and 70%) with different water contents and adding the wood chips of the antrodia camphorata (0.5 g/bottle, 1 g/bottle, 3 g/bottle, 5 g/bottle and 7 g/bottle) into the basic culture media, freeze-drying the culture products, and comparing the dry weight of the antrodia camphorata which can be cultured in each bottle of each group of culture media, which is shown in fig. 1 and fig. 2.
Then, using ursolic acid as a reference substance, and determining the crude triterpene content of the culture product by adopting a vanillin-glacial acetic acid method, so as to select a culture medium with high antrodia camphorata yield and high triterpene component content, which is specifically shown in tables 2 and 3:
TABLE 2 Dry weight and crude triterpene content of Antrodia camphorata on basal medium of different water content (50 d)
Table 3 addition of Dry weight and crude triterpene content of Antrodia camphorata on different Antrodia camphorata wood chip media (60d)
As can be seen from tables 2 and 3, the water content of the culture medium is 70%, the wood dust addition amount of the antrodia camphorata is 1 g/bottle, the dry weight of the obtained antrodia camphorata is highest, and the crude triterpene content of the culture product is the highest.
As shown in FIG. 3, after culturing for a certain period of time, a dense mycelium layer with a thickness of about 3-5cm and capable of being easily peeled off is formed on the surface of the substrate;
as shown in FIG. 4, after sampling and freeze-drying in a sterile super clean bench, 90 days of culture per bottle can obtain a dry weight of about 15g of Antrodia camphorata product.
It should be understood that the above-mentioned embodiments are merely preferred embodiments of the present invention, and not intended to limit the scope of the invention, therefore, all equivalent modifications of the principles of the present invention should be included in the scope of the present invention.
Claims (1)
1. A culture product and substrate completely separated antrodia camphorata culture method is characterized in that: the method mainly comprises the following steps:
(1) activating and inoculating raw materials: taking Antrodia camphorata strain stored at 4 ℃ to perform activated culture on a PDA plate culture medium for 20 days, and punching a hole at a position 0.4cm close to the edge of a mycelium layer to obtain a mycelium block for inoculation;
(2) preparing a solid fermentation medium: weighing a certain amount of cereal raw materials and cinnamomum kanehirae swamp wood chips, placing the cereal raw materials and the cinnamomum kanehirae swamp wood chips in a culture bottle, uniformly mixing the cereal raw materials, placing the cereal raw materials in the lower layer of the culture bottle, and flatly paving the cinnamomum kanehirae swamp wood chips on the lower layer of the culture bottle; mixing glucose and KH2PO4、MgSO4·7H2Dissolving O in a certain amount of distilled water to form a solution containing glucose and inorganic salts, pouring the solution into a culture bottle to soak the solid materials to ensure that the final water content of the culture medium is 50-70%, wherein the soaking time is 2 hours, and then placing the culture medium into an autoclave to be sterilized, wherein the sterilization temperature is 121 ℃ and the sterilization time is 30 minutes;
(3) and (3) punching and inoculating: drilling a cylindrical groove with the depth of 2cm at the center of the solid culture medium by using a glass rod, inoculating one surface of a raw material hypha block containing hypha in the cylindrical groove hole by adhering to a culture medium, and placing in a constant-temperature incubator for dark culture at 28 ℃;
(4) sampling by an aseptic super-clean bench: culturing for a certain time to form a compact mycelium layer with a thickness of 3-5cm and easy stripping on the surface of the substrate, sampling in a sterile ultra-clean bench, and allowing the rest mycelium in the culture medium to continue to grow;
the processing method of the camphor tree wood chips comprises the steps of placing the camphor tree wood chips outdoors for insolation, then sterilizing the camphor tree wood chips by using an autoclave, wherein the sterilization temperature is 121 ℃, the sterilization time is 2 hours, and after sterilization, placing the camphor tree wood chips into an oven for drying at the temperature of 80 ℃;
the grain raw materials comprise 50g of rice and 2.5g of bran, and the dry materials are uniformly mixed and then are put into a culture bottle, wherein the volume of the culture bottle is 450 mL;
the cinnamomum kanehirae wood chips are obtained by crushing cinnamomum kanehirae wood into 0.5 cm-shaped chip wood chips, weighing 1-5g of dried cinnamomum kanehirae wood chips, and flatly paving the dried cinnamomum kanehirae wood chips on the surface of a grain substrate;
in the solution containing glucose and inorganic salt, the content of glucose is 5g, KH2PO4Is 0.5g, MgSO4·7H2O was 0.5 g.
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| CN104904494A (en) * | 2015-05-12 | 2015-09-16 | 柳州市耕青科技有限公司 | Method for manufacturing and cultivating antrodia solid plant bacterium basic seeds |
| CN107586724A (en) * | 2017-08-29 | 2018-01-16 | 殷东林 | A kind of mycelial cultural method of Antrodia camphorata |
| CN108901611A (en) * | 2018-06-14 | 2018-11-30 | 广东食品药品职业学院 | A kind of Antrodia camphorata culture medium and its preparation method and application |
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| CN104928187A (en) * | 2015-05-12 | 2015-09-23 | 柳州市耕青科技有限公司 | Method capable of reducing pollution of antrodia solid plant bacterium protospecies culture medium |
| TW201643243A (en) * | 2015-06-10 | 2016-12-16 | 茁壯企業股份有限公司 | Solid culture medium for culturing fruiting bodies of Antrodia cinnamomea and method for culturing the same |
| CN105052558B (en) * | 2015-09-08 | 2017-12-15 | 福建农林大学 | A kind of fast breeding method of Antrodia camphorata |
| CN107278622A (en) * | 2017-07-11 | 2017-10-24 | 重庆丽妃丹生物科技有限公司 | Antrodia camphorata process for solid culture |
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| CN104904494A (en) * | 2015-05-12 | 2015-09-16 | 柳州市耕青科技有限公司 | Method for manufacturing and cultivating antrodia solid plant bacterium basic seeds |
| CN107586724A (en) * | 2017-08-29 | 2018-01-16 | 殷东林 | A kind of mycelial cultural method of Antrodia camphorata |
| CN108901611A (en) * | 2018-06-14 | 2018-11-30 | 广东食品药品职业学院 | A kind of Antrodia camphorata culture medium and its preparation method and application |
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