Method for increasing content of ganoderma triterpenes in ganoderma lucidum

Abstract

The invention relates to the field of lucid ganoderma cultivation, and particularly discloses a method for increasing the content of ganoderma triterpenes in lucid ganoderma, which comprises the following steps: S1, preparing a culture medium; step S2, basswood manufacturing; step S3, inoculation; step S4, mycelium culture; step S5, burying soil and growing ganoderma lucidum; step S6, forming; from two aspects, firstly, a fermentation accelerant is added during preparation of the culture medium, and the fermentation accelerant is prepared by mixing calcium lactate, salicylic acid, a citrate-phosphate buffer solution and hydroxypropyl-beta-cyclodextrin according to a specific mass ratio; secondly, in the soil burying and ganoderma lucidum growing process, culture medium soil rich in fermentation extract is adopted for culture, the fermentation extract is prepared into a fermentation substrate through compound bacteria fermentation by taking Chinese wolfberry residues, liquorice residues and astragalus membranaceus residues as raw materials, and the content of ganoderma lucidum triterpenes in ganoderma lucidum is increased through the synergistic effect of all the substances.

Description

Technical Field

[0001] The present application relates to the field of Ganoderma lucidum cultivation, and more particularly, to a method for increasing the content of Ganoderma triterpenes in Ganoderma lucidum. Background Art

[0002] Triterpenes are the main active substances in the secondary metabolites of Ganoderma lucidum. They have pharmacological activities such as anti-tumor, antioxidant, liver protection and detoxification, and anti-HIV virus. They are widely used in the pharmaceutical and health care products industries. Therefore, it is of great significance to increase the content of Ganoderma lucidum triterpenes in Ganoderma lucidum fruiting bodies.

[0003] Chinese patent application CN109618812B discloses a method for increasing the triterpenoid content in Ganoderma lucidum. This method uses high-concentration oxygen to induce Ganoderma lucidum to produce more triterpenoids during the fruiting body maturation period. The process is simple and suitable for large-scale application. However, due to the diversion of resources to fruiting body development and environmental pressure during the burial and emergence stage, triterpenoid synthesis will be reduced. In contrast, the triterpenoid content of Ganoderma lucidum can be significantly increased by optimizing metabolic resource allocation, environmental conditions and gene expression during the mycelial culture stage. Therefore, it is particularly important to increase the triterpenoid content in Ganoderma lucidum during the mycelial culture stage.

[0004] Based on the above statements, the present application provides a method for increasing the content of triterpenes in Ganoderma lucidum.

[0005] Application Contents In order to solve the problems mentioned in the above background technology, the present application provides a method for increasing the content of triterpenes in Ganoderma lucidum.

[0006] A method for increasing the content of triterpenes in Ganoderma lucidum, comprising the following steps: Step S1, preparing a culture medium: the culture medium is composed of the following raw materials in parts by weight: 42-46 parts of sawdust, 36-42 parts of cottonseed hulls, 14-16 parts of wheat bran, 0.8-1.2 parts of gypsum, 0.6-1 part of a fermentation promoter, and 266-274 parts of deionized water, and the above raw materials are uniformly mixed by weight to obtain a culture medium; Step S2, basswood production: Cyclobalanopsis glauca is selected as the cultivation material, the Cyclobalanopsis glauca is cut into 10-20 cm basswood, and dried under natural conditions until the moisture content of the Cyclobalanopsis glauca drops to 26-30%, the Cyclobalanopsis glauca and the culture medium are placed in a cultivation bag and then sterilized; Step S3, inoculation: After sterilization, the cultivation bag is inoculated with red Ganoderma lucidum in a sterile environment; Step S4, mycelium cultivation: After inoculation, the cultivation bag is transferred to the culture medium prepared in step S1 for cultivation. During the cultivation process, the temperature is controlled at 22-26° C., the air humidity is maintained at 45-55%, and ventilation is performed 4-6 times a day, each time for 25-35 minutes, until the mycelium covers the surface of the Cyclobalanopsis glauca. Step S5, burying the soil to produce the fungus: remove the bag of the Cyclobalanopsis glauca covered with mycelium and place it in the ridge ditch of the greenhouse, cover the Cyclobalanopsis glauca with the cultivation medium soil, and irradiate the ridge surface with a light source. During the irradiation, the light source is set vertically above the ridge surface, and the air humidity is adjusted to 86-90%. Cultivate for 22-26 days to obtain fruiting bodies; Step S6: Shaping: The Ganoderma lucidum fruiting body is stored in the dark at 3-10°C for 1-5 days.

[0007] Preferably, in step S2, the sterilization temperature is 120-122°C and the sterilization time is 2-3 hours.

[0008] Preferably, in step S5, the temperature of the greenhouse is maintained at 22-26°C, the air humidity is maintained at 75-85%, and the pH of the soil is controlled at pH 5-6.

[0009] Preferably, during the cultivation process in step S5, the light intensity is controlled at 800-1000Lu, the light source is an LED light source, the distance between the LED light source and the fruiting body is 60-90cm, and the LED light source is turned off at night.

[0010] Preferably, in step S5, the cultivation matrix soil is sandy soil with a pH of 5.5-6.5, an organic matter content of not less than 10 g / kg, a porosity of 67.33-80.87%, and a water content of 22.65-59.07%, and is doped with a modified matrix equivalent to 8-12 wt% of the net weight of the soil after exposure to sterilization.

[0011] Preferably, the modified matrix is ​​prepared by mixing fermentation extract, peat, vermiculite and perlite in a mass ratio of 3-5:2-4:0.8-1.2:0.6-0.8.

[0012] Preferably, the fermentation extract is prepared by the following method: Step A1, grinding and sieving wolfberry residue, licorice residue, and astragalus residue together to obtain a mixed powder; controlling the mass ratio of the mixed powder to deionized water to be 1:10-12, adding the mixed powder and deionized water to mix evenly, heating to boiling and extracting for 26-34 minutes, and cooling to obtain a fermentation substrate, wherein the mass ratio of wolfberry residue, licorice residue, and astragalus residue is 12-16:10-14:3-6; Step A2: Add the composite bacteria to the fermentation substrate, stir evenly, ferment for 24-36 hours, centrifuge, collect the supernatant, and spray dry to obtain a fermentation extract.

[0013] Preferably, in step A2, the number of viable bacteria in the fermentation substrate is controlled to be 2.6×10 9 -3.4×10 9 CFU / g.

[0014] Preferably, the composite bacteria comprises Lactobacillus plantarum, Bacillus licheniformis and Paecilomyces chinensis in a live cell count ratio of 1:0.5-0.7:0.2-0.4.

[0015] Preferably, the fermentation promoter is prepared by the following steps: Calcium lactate and salicylic acid are added to a citrate-phosphate buffer solution, and hydroxypropyl-β-cyclodextrin is added and stirred evenly to obtain a fermentation promoter, wherein the mass ratio of calcium lactate, salicylic acid, citrate-phosphate buffer solution and hydroxypropyl-β-cyclodextrin is 0.06-0.08:0.02-0.04:26-34:1-1.4.

[0016] Compared with the prior art, this application has the following beneficial effects: In order to increase the content of triterpenoids in Ganoderma lucidum, the technical solution of the present invention starts from two aspects. First, a fermentation promoter is added to the culture medium. The fermentation promoter is prepared by mixing calcium lactate, salicylic acid, citrate-phosphate buffer and hydroxypropyl-β-cyclodextrin in a specific mass ratio. Calcium lactate can not only provide calcium ions Ca 2+, activates the calcium signaling pathway in Ganoderma cells, upregulates the expression of key enzyme genes for triterpene synthesis, and promotes metabolic flow towards triterpene synthesis. Moreover, it can also participate in the tricarboxylic acid cycle, provide precursors for triterpene synthesis, and promote the synthesis of Ganoderma triterpenes; the presence of salicylic acid can induce the accumulation of reactive oxygen species, triggering the defense response of Ganoderma lucidum, thereby promoting the synthesis of secondary metabolites; citrate-phosphate buffer can not only stabilize the pH in an appropriate range by regulating the acid-base balance, avoiding the inhibition of triterpene synthase activity by extreme pH, but it can also serve as a carbon source to synergistically promote the synthesis of Ganoderma triterpenes; hydroxypropyl-β-cyclodextrin can not only enclose Ganoderma triterpenes through the cavity structure to form a stable inclusion complex, reducing the triterpenes in the culture medium Degradation or volatilization, thereby relieving feedback inhibition and promoting the continuous synthesis of triterpenes by the bacteria. It can also change the cell membrane structure and promote the secretion or accumulation of triterpenes. Secondly, in the process of burying the ganoderma, cultivation substrate soil rich in fermentation extracts is used for cultivation. The fermentation extracts are made of wolfberry residue, licorice residue and astragalus residue through composite bacteria fermentation to make fermentation substrates. By decomposing the complex polysaccharides in the traditional Chinese medicine residues, fermentable sugars are provided as a carbon source for the growth of Ganoderma lucidum, promoting the synthesis of triterpenes. The vitamins and amino acids produced by the decomposition of Bacillus licheniformis can promote the key enzymes for the synthesis of Ganoderma lucidum triterpenes. Chinese pseudo-penicillium can decompose the crude fiber in the traditional Chinese medicine residues and release active ingredients such as flavonoids and saponins. Through the above effects, the content of Ganoderma lucidum triterpenes in Ganoderma lucidum is synergistically improved. DETAILED DESCRIPTION

[0017] In order to make the implementation methods of the present application easier to understand, the present application will be described in detail below with reference to specific examples. These examples are only for illustration and are not intended to limit the scope of application of the present application.

[0018] The contents of the main raw materials and their components used in the examples and comparative examples are as follows: The Ganoderma lucidum strain was provided and preserved by the Shanghai Edible Fungi Branch of the Agricultural Microbiology Center of the China Culture Collection Administration, with the strain number: Ganoderma lucidum G0023; the sandy soil (2-6 mm) was provided by Chongqing Maidiling Agriculture Co., Ltd.; the preservation number of Lactobacillus plantarum is CGMCC 18401, which is from the China Center for Type Culture Collection, the preservation number of Paecilomyces chinensis is CICC 10098, and the preservation number of Bacillus licheniformis is CICC 10098, which is from the China Center for Industrial Microbiology Culture Collection.

[0019] Table 1 Nutrient content (dry matter basis) % Preparation Examples 1-3 and Comparative Preparation Examples 1-2 provide a fermentation accelerator.

[0020] Preparation Example 1 This preparation example provides a fermentation promoter, which is prepared by the following steps: Calcium lactate and salicylic acid are added to a citrate-phosphate buffer solution, and then hydroxypropyl-β-cyclodextrin is added and stirred at a rotation speed of 560 rpm for 12 minutes until uniform to obtain a fermentation promoter, wherein the mass ratio of calcium lactate, salicylic acid, citrate-phosphate buffer solution and hydroxypropyl-β-cyclodextrin is 0.06:0.02:26:1.

[0021] Preparation Example 2 This preparation example provides a fermentation promoter, which is prepared by the following steps: Calcium lactate and salicylic acid are added to a citrate-phosphate buffer solution, and then hydroxypropyl-β-cyclodextrin is added and stirred at a rotation speed of 580 rpm for 14 minutes until uniform to obtain a fermentation promoter, wherein the mass ratio of calcium lactate, salicylic acid, citrate-phosphate buffer solution and hydroxypropyl-β-cyclodextrin is 0.07:0.03:30:1.2.

[0022] Preparation Example 3 This preparation example provides a fermentation promoter, which is prepared by the following steps: Calcium lactate and salicylic acid are added to a citrate-phosphate buffer solution, and then hydroxypropyl-β-cyclodextrin is added and stirred uniformly to obtain a fermentation promoter, wherein the mass ratio of calcium lactate, salicylic acid, citrate-phosphate buffer solution and hydroxypropyl-β-cyclodextrin is 0.08:0.04:34:1.4.

[0023] Comparative Preparation Example 1 This comparative preparation example provides a fermentation accelerator, which is prepared by the following steps: Calcium lactate and salicylic acid are added to deionized water, and then hydroxypropyl-β-cyclodextrin is added and stirred at a rotation speed of 560 rpm for 12 minutes until uniform to obtain a fermentation promoter, wherein the mass ratio of calcium lactate, salicylic acid, citrate-phosphate buffer and hydroxypropyl-β-cyclodextrin is 0.06:0.02:26:1.

[0024] Comparative Preparation Example 2 This comparative preparation example provides a fermentation accelerator, which is prepared by the following steps: Calcium lactate and salicylic acid are added to a citrate-phosphate buffer solution, and then glucose is added. The mixture is stirred at a rotation speed of 560 rpm for 12 minutes until uniform, to obtain a fermentation promoter, wherein the mass ratio of calcium lactate, salicylic acid, citrate-phosphate buffer solution and hydroxypropyl-β-cyclodextrin is 0.06:0.02:26:1.

[0025] Preparation Examples 4-6 and Comparative Preparation Examples 3-4 provide a fermentation extract.

[0026] Preparation Example 4 This preparation example provides a fermentation extract, which is prepared by the following steps: Step A1: Grind wolfberry residue, licorice residue, and astragalus residue together and pass through a 120-mesh sieve to obtain a mixed powder; control the mass ratio of the mixed powder to deionized water to be 1:10, stir the mixed powder with deionized water at a speed of 610 rpm for 14 minutes until uniformly mixed, heat to boiling and extract for 26 minutes, and cool to obtain a fermentation substrate, wherein the mass ratio of wolfberry residue, licorice residue, and astragalus residue is 12:10:3; Step A2: Add composite bacteria to the fermentation substrate to control the number of viable bacteria in the fermentation substrate to 2.6×10 9 CFU / g, control the rotation speed to 520rpm, stir for 12min until uniform, ferment for 24h, centrifuge, take the supernatant, spray dry at 70℃ to obtain the fermentation extract, wherein the composite bacteria include Lactobacillus plantarum, Bacillus licheniformis and Paecilomyces sinensis with a viable cell count ratio of 1:0.5:0.2.

[0027] Preparation Example 5 This preparation example provides a fermentation extract, which is prepared by the following steps: Step A1: Grind wolfberry residue, licorice residue, and astragalus residue together and pass through a 140-mesh sieve to obtain a mixed powder; control the mass ratio of the mixed powder to deionized water to be 1:11, stir the mixed powder with deionized water at 630 rpm for 16 minutes until uniformly mixed, heat to boiling and extract for 30 minutes, and cool to obtain a fermentation substrate, wherein the mass ratio of wolfberry residue, licorice residue, and astragalus residue is 14:12:4.5; Step A2: Add composite bacteria to the fermentation substrate to control the number of viable bacteria in the fermentation substrate to 3.0×10 9 CFU / g, control the rotation speed to 540 rpm, stir for 14 min until uniform, ferment for 30 h, centrifuge, take the supernatant, spray dry at 75 ° C to obtain a fermentation extract, wherein the composite bacteria include Lactobacillus plantarum, Bacillus licheniformis and Paecilomyces sinensis with a viable cell count ratio of 1:0.6:0.3.

[0028] Preparation Example 6 This preparation example provides a fermentation extract, which is prepared by the following steps: Step A1: Grind wolfberry residue, licorice residue, and astragalus residue together and pass through a 160-mesh sieve to obtain a mixed powder; control the mass ratio of the mixed powder to deionized water to be 1:12, stir the mixed powder with deionized water at 650 rpm for 18 minutes until uniformly mixed, heat to boiling and extract for 34 minutes, and cool to obtain a fermentation substrate, wherein the mass ratio of wolfberry residue, licorice residue, and astragalus residue is 16:14:6; Step A2: Add composite bacteria to the fermentation substrate to control the number of viable bacteria in the fermentation substrate to 3.4×10 9 CFU / g, the rotation speed was controlled at 560 rpm, stirred for 16 min until uniform, fermented for 36 h, centrifuged, the supernatant was taken, and spray-dried at 80 ° C to obtain a fermentation extract, wherein the composite bacteria included Lactobacillus plantarum, Bacillus licheniformis and Paecilomyces sinensis with a viable cell count ratio of 1:0.7:0.4.

[0029] Comparative Preparation Example 3 This comparative preparation example provides a fermentation extract, which is prepared by the following steps: Step A1: Grind wolfberry residue, licorice residue, and astragalus residue together and pass through a 120-mesh sieve to obtain a mixed powder; control the mass ratio of the mixed powder to deionized water to be 1:10, stir the mixed powder with deionized water at a speed of 610 rpm for 14 minutes until uniformly mixed, heat to boiling and extract for 26 minutes, and cool to obtain a fermentation substrate, wherein the mass ratio of wolfberry residue, licorice residue, and astragalus residue is 12:10:3; Step A2: Add composite bacteria to the fermentation substrate to control the number of viable bacteria in the fermentation substrate to 2.6×10 9 CFU / g, control the rotation speed to 520rpm, stir for 12min until uniform, ferment for 24h, centrifuge, take the supernatant, spray dry at 70℃ to obtain the fermentation extract, wherein the composite bacteria include Lactobacillus plantarum and Paecilomyces sinensis with a live bacteria ratio of 1:0.7.

[0030] Comparative Preparation Example 4 This comparative preparation example provides a fermentation extract, which is prepared by the following steps: Step A1: Grind wolfberry residue, licorice residue, and astragalus residue together and pass through a 120-mesh sieve to obtain a mixed powder; control the mass ratio of the mixed powder to deionized water to be 1:10, stir the mixed powder with deionized water at a speed of 610 rpm for 14 minutes until uniformly mixed, heat to boiling and extract for 26 minutes, and cool to obtain a fermentation substrate, wherein the mass ratio of wolfberry residue, licorice residue, and astragalus residue is 12:10:3; Step A2: Add composite bacteria to the fermentation substrate to control the number of viable bacteria in the fermentation substrate to 2.6×10 9 CFU / g, control the rotation speed to 520rpm, stir for 12min until uniform, ferment for 24h, centrifuge, take the supernatant, spray dry at 70℃ to obtain the fermentation extract, wherein the composite bacteria include Lactobacillus plantarum and Bacillus licheniformis with a live cell count ratio of 1:0.7.

[0031] Examples 1-3 and Comparative Examples 1-4 provide a method for increasing the content of triterpenes in Ganoderma lucidum.

[0032] Example 1 This embodiment provides a method for increasing the content of triterpenes in Ganoderma lucidum, comprising the following steps: Step S1, preparing a culture medium: the culture medium is composed of the following raw materials in parts by weight: 42 parts of sawdust, 36 parts of cottonseed hulls, 14 parts of wheat bran, 0.8 parts of gypsum, 0.6 parts of the fermentation promoter prepared in Preparation Example 1, and 266 parts of deionized water. The above raw materials are mixed uniformly by weight to obtain a culture medium; Step S2, basswood production: Cyclobalanopsis glauca is selected as the cultivation material, the Cyclobalanopsis glauca is cut into 10 cm basswood pieces, and dried under natural conditions until the moisture content of the Cyclobalanopsis glauca is reduced to 25%. The Cyclobalanopsis glauca and the culture medium are placed in a cultivation bag and then sterilized at a temperature of 120° C. for 2 hours. Step S3, inoculation: After sterilization, the cultivation bag is inoculated with red Ganoderma lucidum in a sterile environment; Step S4, mycelium cultivation: After inoculation, the cultivation bag is transferred to the culture medium prepared in step S1 for cultivation. During the cultivation process, the temperature is controlled at 22° C., the air humidity is maintained at 45%, and ventilation is performed four times a day, each time for 25 minutes, until the mycelium covers the surface of the Cyclobalanopsis glauca. Step S5, burying the soil to produce mushrooms: remove the bag of the cycad covered with mycelium and place it in the ridge of the greenhouse, cover the cycad with the cultivation matrix soil, and irradiate the ridge surface with a light source. During the irradiation, the light source is vertically set directly above the ridge surface, and the air humidity is adjusted to 86%. Cultivate for 22 days to obtain fruiting bodies, wherein the temperature of the greenhouse is maintained at 22°C, the air humidity is maintained at 75%, the pH of the soil is controlled at pH 5, the cultivation matrix soil is a sandy soil with a pH of 5.5, an organic matter content of not less than 10g / kg, a porosity of 67.33%, and a water content of 22.65%. After exposure to the sun for sterilization and disinfection, a modified matrix is ​​added in a weight ratio equivalent to 8wt% of the net weight of the soil, and the modified matrix is ​​prepared by mixing the fermentation extract prepared in Preparation Example 4, peat, vermiculite and perlite in a mass ratio of 3:2:0.8:0.6; Step S6, forming: storing the Ganoderma lucidum fruiting body in the dark at 3° C. for 1 day.

[0033] Example 2 This embodiment provides a method for increasing the content of triterpenes in Ganoderma lucidum, comprising the following steps: Step S1, preparing a culture medium: the culture medium is composed of the following raw materials in parts by weight: 44 parts of sawdust, 39 parts of cottonseed hulls, 15 parts of wheat bran, 1 part of gypsum, 0.8 parts of the fermentation promoter prepared in Preparation Example 2, and 270 parts of deionized water. The above raw materials are mixed uniformly by weight to obtain a culture medium; Step S2, basswood production: Cyclobalanopsis glauca is selected as the cultivation material, the Cyclobalanopsis glauca is cut into 15 cm basswood, and dried under natural conditions until the moisture content of the Cyclobalanopsis glauca is reduced to 28%. The Cyclobalanopsis glauca and the culture medium are placed in a cultivation bag and then sterilized at a temperature of 121° C. for 2.5 hours. Step S3, inoculation: After sterilization, the cultivation bag is inoculated with red Ganoderma lucidum in a sterile environment; Step S4, mycelium cultivation: After inoculation, the cultivation bag is transferred to the culture medium prepared in step S1 for cultivation. During the cultivation process, the dark is maintained, the temperature is controlled at 24° C., the air humidity is maintained at 50%, and ventilation is performed 5 times a day, each time for 30 minutes, until the mycelium covers the surface of the Cyclobalanopsis glauca. Step S5, burying the soil to produce mushrooms: remove the bag of the cycad covered with mycelium and place it in the ridge of the greenhouse, cover the cycad with the cultivation matrix soil, and irradiate the ridge surface with a light source. During the irradiation, the light source is vertically set directly above the ridge surface, and the air humidity is adjusted to 88%. Cultivate for 24 days to obtain fruiting bodies, wherein the temperature of the greenhouse is maintained at 24°C, the air humidity is maintained at 80%, the pH value of the soil is controlled at pH 5.5, the cultivation matrix soil is a sandy soil with a pH value of 6.0, an organic matter content of not less than 10 g / kg, a porosity of 74.1%, and a water content of 40.86%. After exposure to the sun for sterilization and disinfection, the modified matrix is ​​doped with a weight ratio equivalent to 10 wt% of the net weight of the soil, and the modified matrix is ​​prepared by mixing the fermentation extract prepared in Preparation Example 5, peat, vermiculite and perlite in a mass ratio of 4:3:1:0.7; Step S6, forming: storing the Ganoderma lucidum fruiting body in the dark at 6° C. for 3 days.

[0034] Example 3 This embodiment provides a method for increasing the content of triterpenes in Ganoderma lucidum, comprising the following steps: Step S1, preparing a culture medium: the culture medium is composed of the following raw materials in parts by weight: 46 parts of sawdust, 42 parts of cottonseed hulls, 16 parts of wheat bran, 1.2 parts of gypsum, 1 part of the fermentation promoter prepared in Preparation Example 3, and 274 parts of deionized water. The above raw materials are mixed uniformly by weight to obtain a culture medium; Step S2, basswood production: Cyclobalanopsis glauca is selected as the cultivation material, the Cyclobalanopsis glauca is cut into 20 cm basswood pieces, and dried under natural conditions until the moisture content of the Cyclobalanopsis glauca is reduced to 30%. The Cyclobalanopsis glauca and the culture medium are placed in a cultivation bag and then sterilized at a controlled sterilization temperature of 122° C. for 3 hours. Step S3, inoculation: After sterilization, the cultivation bag is inoculated with red Ganoderma lucidum in a sterile environment; Step S4, mycelium cultivation: After inoculation, the cultivation bag is transferred to the culture medium prepared in step S1 for cultivation. During the cultivation process, the darkness is maintained, the temperature is controlled at 26° C., the air humidity is maintained at 55%, and ventilation is performed 6 times a day, each time for 35 minutes, until the mycelium covers the surface of the Cyclobalanopsis glauca. Step S5, burying the soil to produce mushrooms: remove the bag of the cycad covered with mycelium and place it in the ridge of the greenhouse, cover the cycad with the cultivation matrix soil, and irradiate the ridge surface with a light source. During the irradiation, the light source is vertically set directly above the ridge surface, and the air humidity is adjusted to 90%. Cultivate for 26 days to obtain fruiting bodies, wherein the temperature of the greenhouse is maintained at 26°C, the air humidity is maintained at 85%, the pH value of the soil is controlled at pH 6, the cultivation matrix soil is a sandy soil with a pH value of 5.5, an organic matter content of not less than 10 g / kg, a porosity of 80.87%, and a water content of 59.07%. After exposure to the sun for sterilization and disinfection, the modified matrix is ​​doped with a weight ratio equivalent to 12 wt% of the net weight of the soil, and the modified matrix is ​​prepared by mixing the fermentation extract prepared in Preparation Example 6, peat, vermiculite and perlite in a mass ratio of 5:4:1.2:0.8; Step S6, forming: storing the Ganoderma lucidum fruiting body in the dark at 10° C. for 5 days.

[0035] Comparative Example 1 Comparative Example 1 is the same as Example 1, except that the fermentation promoter in Example 1 is replaced by the fermentation promoter prepared in Comparative Preparation Example 1.

[0036] Comparative Example 2 Comparative Example 2 is the same as Example 1, except that the fermentation promoter in Example 1 is replaced by the fermentation promoter prepared in Comparative Preparation Example 2.

[0037] Comparative Example 3 Comparative Example 3 is the same as Example 1, except that the fermentation extract in Example 1 is replaced by the fermentation extract prepared in Comparative Preparation Example 3.

[0038] Comparative Example 4 Comparative Example 4 is the same as Example 1, except that the fermentation extract in Example 1 is replaced by the fermentation extract prepared in Comparative Preparation Example 4.

[0039] Performance Testing Take 0.2 g of Ganoderma lucidum dry powder, add anhydrous ethanol to 10 mL, ultrasonically disrupt for 2 h, shake well, centrifuge at 4000 g for 10 min, and retain the supernatant. Take 0.1 mL of the supernatant, add 0.2 mL of 5% vanillin-glacial acetic acid solution and 0.5 mL of perchloric acid solution, shake well, heat at 60°C for 20 min, remove and immediately place in an ice bath to cool for 10 min, add 5 mL of glacial acetic acid, and measure the absorbance of the reaction solution at 550 nm. Ethanol is used as a blank control. The formula for calculating the triterpene content of Ganoderma lucidum is shown in the following formula (1): (1) Wherein, C (µg): triterpene content of sample decomposition solution obtained from the standard curve; V1 (mL): sample fixed volume; V2 (mL): sample decomposition solution volume; m (g): sample mass. The test results are shown in Table 1 below. Table 2 Effects of different treatments on the triterpenoid content of Ganoderma lucidum in Examples 1-3 and Comparative Examples 1-4 As shown in Table 1, compared with Comparative Examples 1-4, the method provided by Examples 1-3 can better increase the content of triterpenes in Ganoderma lucidum.

[0040] This specific embodiment is merely an explanation of the present application and is not a limitation of the present application. After reading this specification, those skilled in the art may make non-creative modifications to the present embodiment as needed, but as long as they are within the scope of the claims of the present application, they are protected by the patent law.

Claims

1. A method for increasing the content of triterpenoids in Ganoderma lucidum, characterized in that: The following steps are involved: Step S1, preparing a culture medium: the culture medium is composed of the following raw materials in parts by weight: 42-46 parts of sawdust, 36-42 parts of cottonseed hulls, 14-16 parts of wheat bran, 0.8-1.2 parts of gypsum, 0.6-1 part of a fermentation promoter, and 266-274 parts of deionized water, and the above raw materials are uniformly mixed by weight to obtain a culture medium; Step S2, basswood production: cutting the Cyclobalanopsis glauca into 10-20 cm basswood, drying, and placing the Cyclobalanopsis glauca and culture medium in a cultivation bag and then sterilizing; Step S3, inoculation: After sterilization, the cultivation bag is inoculated with red Ganoderma lucidum in a sterile environment; Step S4, mycelium culture: after inoculation, the cultivation bag is transferred to the culture medium prepared in step S1 for culture; Step S5, burying the soil to produce fungi: remove the bags of the Cyclobalanopsis glauca covered with mycelium and place them in the ridges of the greenhouse, cover the Cyclobalanopsis glauca with the cultivation medium soil, illuminate the ridge surface with a light source, and cultivate to obtain fruiting bodies; Step S6: Shaping: The Ganoderma lucidum fruiting body is stored in the dark at 3-10°C for 1-5 days.

2. The method for increasing the triterpenoid content in Ganoderma lucidum according to claim 1, characterized in that: In step S1, the fermentation promoter is prepared by the following steps: Calcium lactate and salicylic acid are added to a citrate-phosphate buffer solution, and then hydroxypropyl-β-cyclodextrin is added and stirred evenly to obtain a fermentation promoter.

3. The method for increasing the triterpenoid content in Ganoderma lucidum according to claim 2, characterized in that: The mass ratio of the calcium lactate, salicylic acid, citrate-phosphate buffer and hydroxypropyl-β-cyclodextrin is 0.06-0.08:0.02-0.04:26-34:1-1.

4.

4. The method for increasing the triterpenoid content in Ganoderma lucidum according to claim 1, characterized in that: In step S4, during the culture process, the culture is kept in darkness, the temperature is controlled at 22-26° C., the air humidity is maintained at 45-55%, and ventilation is performed 4-6 times a day, each time for 25-35 minutes, until the mycelium covers the surface of the Cyclobalanopsis glauca.

5. The method for increasing the triterpenoid content in Ganoderma lucidum according to claim 1, characterized in that: In step S5, during irradiation, the light source is vertically positioned directly above the ridge surface, and the air humidity is adjusted to 86-90%, and the cultivation is carried out for 22-26 days.

6. The method for increasing the triterpenoid content in Ganoderma lucidum according to claim 1, characterized in that: In step S5, the temperature of the greenhouse is maintained at 22-26° C., the air humidity is maintained at 75-85%, and the pH of the soil is controlled at pH 5-6.

7. The method for increasing the triterpenoid content in Ganoderma lucidum according to claim 1, characterized in that: During the cultivation process in step S5, the light intensity is controlled at 800-1000 Lu, the light source is an LED light source, the distance between the LED light source and the fruiting body is 60-90 cm, and the LED light source is turned off at night.

8. The method for increasing the triterpenoid content in Ganoderma lucidum according to claim 1, characterized in that: In step S5, the cultivation substrate soil is sandy soil with a pH of 5.5-6.5, an organic matter content of not less than 10 g / kg, a porosity of 67.33-80.87%, and a water content of 22.65-59.07%. After exposure to sunlight for sterilization and disinfection, the modified substrate is doped with a weight ratio equivalent to 8-12 wt% of the net weight of the soil, and the modified substrate is mixed by fermentation extract, peat, vermiculite and perlite in a mass ratio of 3-5:2-4:0.8-1.2:0.6-0.

8.

9. The method for increasing the triterpenoid content in Ganoderma lucidum according to claim 8, characterized in that: The fermentation extract is prepared by the following method: Step A1: Grind and sieve wolfberry residue, licorice residue, and astragalus residue together to obtain a mixed powder; control the mass ratio of the mixed powder to deionized water to be 1:10-12, mix the mixed powder and deionized water evenly, heat to boiling and extract for 26-34 minutes, and cool to obtain a fermentation substrate; Step A2: Add the composite bacteria to the fermentation substrate, stir evenly, ferment for 24-36 hours, centrifuge, collect the supernatant, and spray dry to obtain a fermentation extract.

10. The method for increasing the triterpenoid content in Ganoderma lucidum according to claim 9, characterized in that: In step A1, the mass ratio of wolfberry residue, liquorice residue and astragalus residue is 12-16:10-14:3-6. In step A2, the number of viable bacteria in the fermentation substrate is 2.6×10 9 -3.4×10 9 CFU / g.

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