Method for culturing palmosa palmosa by utilizing cordyceps militaris cultivation residues

Abstract

The invention provides a method for culturing rose palmatum by utilizing cordyceps militaris cultivation residues, and relates to the technical field of microorganisms. According to the method, the cordyceps militaris cultivation residues are applied to the rose palmata cultivation process for the first time, a new resource recycling method is provided for the cordyceps militaris cultivation residues, the problems of resource waste and environmental pollution of the cordyceps militaris cultivation residues are solved, a new low-cost high-quality cultivation resource is provided for the rose palmata, and the method is suitable for large-scale popularization and application. The problem that the culture cost of the palmarosa palmosa is high is solved. According to the method, a culture method is provided for the palm-shaped rose, rapid culture of the palm-shaped rose is achieved, 35-day bag filling can be achieved, the fastest culture period can reach 47 days, the palm-shaped rose cultivated through the method further has the high sporocarp yield, the highest sporocarp yield can reach 81.2%, and the cultivated palm-shaped rose is good in hypha growth vigor, good in quality and high in yield. The product contains a plurality of antioxidant active substances, such as total polysaccharides, flavones and polyphenols, and has strong antioxidant ability.

Description

Technical Field

[0001] The present application relates to the field of microbial technology, and in particular to a method for culturing palmate rose fungus using Cordyceps militaris cultivation residues. Background Art

[0002] Cordyceps militaris, also known as northern cordyceps, is one of the rare edible and medicinal fungi. It is rich in nutrients and functional ingredients such as protein, amino acids, cordycepin, polyphenols, etc., and has been approved as a new food raw material. Studies have shown that the residues of Cordyceps militaris cultivation are rich in nutrients such as protein and starch, but due to the lack of resource utilization technology, they are usually used as fuel and discarded in the surrounding areas of the production site. Only a small part is purchased by small farms at a low price. Therefore, the resource utilization of Cordyceps militaris cultivation residues is imminent.

[0003] Palmate rose fungus, also known as net-covered red pleated parasol, belongs to the family Agaricaceae and the genus Pleurotus. Its fruiting body is pink and gelatinous. It is rich in polysaccharides, polyphenols and other effective ingredients. It has the effects of protecting the liver and moisturizing the intestines, anti-oxidation, and improving immunity. It has great ornamental and nutritional health value. At present, palmate rose fungus has been cultivated artificially. The palmate rose fungus fruiting body has a smooth taste and a unique fruity aroma. It has excellent color, aroma, taste and shape, and has a broad consumer market. However, the artificial large-scale cultivation of palmate rose fungus is still in its infancy, and there are few reports on the growth characteristics of palmate rose fungus.

[0004] There are no reports on the cultivation technology of Rosa palmata using Cordyceps militaris cultivation residues instead of sawdust and other raw materials. Summary of the invention

[0005] In order to solve the above technical problems, the present invention provides a culture medium for culturing palmate rose fungus using Cordyceps militaris cultivation residues, a culture method and application thereof. On the one hand, by optimizing the palmate rose fungus mother strain, liquid strain culture medium and culture conditions, the growth vitality of palmate rose fungus mycelium is promoted and the strain culture cycle is shortened. On the other hand, the cultivation matrix is ​​innovated with Cordyceps militaris cultivation residues as the main raw material, the cultivation formula is simplified, the production cost is reduced, and the biological efficiency is improved. At the same time, the content of the effective components of the palmate rose fungus obtained by culture and its antioxidant activity correlation are investigated, and the palmate rose fungus fruiting bodies rich in physiologically active substances such as flavonoids and polyphenols are obtained, which can be used as raw materials for food and health products.

[0006] On the one hand, the present application provides the use of Cordyceps militaris cultivation residues in cultivating palmate rose fungus.

[0007] Furthermore, the Cordyceps militaris cultivation residue is obtained by culturing Cordyceps militaris in a Cordyceps militaris culture medium; preferably, the Cordyceps militaris culture medium comprises wheat grains, potassium dihydrogen phosphate and / or magnesium sulfate.

[0008] Those skilled in the art will appreciate that a common Cordyceps militaris culture medium may be selected to culture Cordyceps militaris, thereby obtaining Cordyceps militaris cultivation residues.

[0009] In a preferred embodiment, the Cordyceps militaris culture medium includes wheat grains and / or nutrient solution, which are prepared according to a mass ratio of 1:(1.6-1.7). Preferably, the nutrient solution includes: 1.0-2.0 g of potassium dihydrogen phosphate, 0.5-1.0 g of magnesium sulfate, and 1000 mL of water.

[0010] Among them, the cultivation of Cordyceps militaris can be carried out by general methods. The specific palmate rose ear varieties, Cordyceps militaris varieties and Cordyceps militaris cultivation methods have no effect on the technical effects of this application and are not excessively limited in this application.

[0011] In this application, for the first time, Cordyceps militaris cultivation residues are applied to the palm-shaped rose ear cultivation process, providing a new resource recycling method for Cordyceps militaris cultivation residues, solving the resource waste and environmental pollution problems of Cordyceps militaris cultivation residues, and in this application, it is verified that the use of Cordyceps militaris cultivation residues to cultivate palm-shaped rose ears can improve the growth characteristics of palm-shaped rose ears, shorten the bag-filling time, and increase the fruiting body yield of palm-shaped rose ears, providing a high-quality new culture medium component for palm-shaped rose ears. In addition, this application also proves that the use of Cordyceps militaris cultivation residues to cultivate palm-shaped rose ears can effectively increase the content of antioxidant active substances in palm-shaped rose ears, such as polysaccharides, flavonoids, and polyphenols, thereby improving the antioxidant activity and hydroxyl radical scavenging activity of palm-shaped rose ears.

[0012] On the other hand, the present application also provides a palmate rose ear cultivation medium, which includes Cordyceps militaris cultivation residues.

[0013] Furthermore, the Cordyceps militaris cultivation residue is obtained by culturing Cordyceps militaris in a Cordyceps militaris culture medium; preferably, the Cordyceps militaris culture medium comprises wheat grains, potassium dihydrogen phosphate and / or magnesium sulfate.

[0014] In a preferred embodiment, the Cordyceps militaris culture medium includes wheat grains and / or nutrient solution, which are prepared according to a mass ratio of 1:(1.6-1.7). Preferably, the nutrient solution includes: 1.0-2.0 g of potassium dihydrogen phosphate, 0.5-1.0 g of magnesium sulfate, and 1000 mL of water.

[0015] Furthermore, the content of the Cordyceps militaris cultivation residue is 200-600 parts by mass of the culture medium; preferably, 200-500 parts; more preferably, 500 parts.

[0016] The content of the Cordyceps militaris cultivation residue may be any value among 200 parts, 300 parts, 400 parts, 500 parts and 600 parts.

[0017] Further, the culture medium further comprises sawdust and / or quicklime; preferably, 400-800 parts of sawdust; preferably, 8-15 parts of quicklime; more preferably, 500 parts of sawdust; more preferably, 10 parts of quicklime;

[0018] And / or, the culture medium further comprises water; preferably, the water content of the culture medium is 55%-60% by mass percentage of the culture medium.

[0019] In a preferred embodiment, the palmate rose fungus cultivation medium includes 500g of sawdust, 500g of Cordyceps militaris cultivation residue, 10g of quicklime, and water is added to make the water content in the culture material reach 55%-60%.

[0020] Those skilled in the art will appreciate that the cultivation medium for the palmate rose ear can be prepared by a general method.

[0021] In a preferred embodiment, the method for preparing the palmate rose ear cultivation medium comprises:

[0022] Step 1: Soak the Cordyceps militaris cultivation residue in water for 3-4 hours, drain the water and set aside;

[0023] Step 2: Add water to the Cordyceps militaris cultivation residues, sawdust and quicklime and mix well.

[0024] Preferably, the culture medium further comprises a high temperature sterilization step, wherein the culture medium is sterilized at 121° C.-125° C. for 2 hours.

[0025] On the other hand, the present application also provides a method for culturing palmate rose fungus using Cordyceps militaris cultivation residues, the method comprising: cultivating palmate rose fungus using the palmate rose fungus cultivation medium.

[0026] Furthermore, the method comprises the following steps:

[0027] Step 1: Inoculate the palmate rose fungus into the palmate rose fungus cultivation medium at an inoculation volume of 10-15 mL / bag, and perform germ management in a light-proof environment at 18°C-22°C and a relative humidity of less than 70%;

[0028] Step 2: Perform bud-inducing management under the conditions of 300-500 Lux diffuse light for 12-16 hours per day, carbon dioxide concentration of 800-1000 ppm, humidity of 80%-90%, and 16°C-22°C;

[0029] Step 3: Carry out mushroom production management under the conditions of 300-500 Lux diffuse light for 12-16 hours per day, carbon dioxide concentration of 800-1000 ppm, humidity of 80%-90%, and 20°C-22°C.

[0030] In a specific embodiment, during the spawn management, bud inducing management can be carried out when the hyphae spread and grow throughout the culture medium; fruiting body culture can be carried out when white to light pink palmate rose ear primordia appear.

[0031] Preferably, the method further comprises the step of inoculating palmate rose ear into palmate rose ear mother seed culture medium to obtain palmate rose ear mother seed; more preferably, the palmate rose ear mother seed culture medium comprises: a carbon source and / or a nitrogen source; more preferably, the carbon source is selected from one or more of sucrose, lactose, glucose, maltose, and mannitol; more preferably, the carbon source is maltose; more preferably, the nitrogen source is selected from one or more of bean powder, cornmeal, malt powder, wheat bran, ammonium sulfate, ammonium chloride, peanut cake powder, and wheat grain powder; more preferably, the nitrogen source is peanut cake powder; more preferably, the palmate rose ear mother seed culture medium further comprises: potassium dihydrogen phosphate, magnesium sulfate and / or vitamin B1; more preferably, the palmate rose ear mother seed culture medium comprises: 1%-5% maltose, 1%-5% peanut cake powder, 0.1%-0.5% potassium dihydrogen phosphate, 0.1%-0.5% magnesium sulfate, vitamin B1 0.001%-0.005%; more preferably, the palmate rose mother culture medium includes: 2% maltose, 1.5% peanut powder, 0.2% potassium dihydrogen phosphate, 0.1% magnesium sulfate, and 0.002% vitamin B1; preferably, the culture conditions include: 16°C-28°C, pH4-9; more preferably, 22°C-25°C, pH7-9, 15-20d.

[0032] More preferably, 22°C, pH7.

[0033] In a preferred embodiment, the palmate rose mother culture medium comprises: 20 g maltose, 15 g peanut powder, 2 g potassium dihydrogen phosphate, 1 g magnesium sulfate, 20 mg vitamin B1, 20 g agar, 1000 mL water, pH 7.0.

[0034] Preferably, the method further comprises the step of inoculating the palmate rose ear mother seed into the palmate rose ear liquid stock culture medium to obtain the palmate rose ear liquid stock; more preferably, the palmate rose ear liquid stock culture medium comprises: a carbon source and / or a nitrogen source; more preferably, the carbon source is selected from one or more of sucrose, lactose, glucose, maltose, and mannitol; more preferably, the carbon source is maltose; more preferably, the nitrogen source is selected from one or more of bean powder, cornmeal, malt powder, wheat bran, ammonium sulfate, ammonium chloride, peanut cake powder, and wheat grain powder; more preferably, the nitrogen source is peanut cake powder; more preferably, the palmate rose ear liquid stock culture medium further comprises: potassium dihydrogen phosphate, magnesium sulfate and / or vitamin B1; more preferably, the palmate rose ear liquid stock culture medium comprises: 1%-5% maltose, 1%-5% peanut cake powder, 0.1%-0.5% potassium dihydrogen phosphate, 0.1%-0.5% magnesium sulfate, vitamin B1 0.001%-0.005%; more preferably, the palmate rose ear liquid stock culture medium includes: 3% maltose, 1% peanut powder, 0.2% potassium dihydrogen phosphate, 0.1% magnesium sulfate, and 0.002% vitamin B1; preferably, the culture conditions include: pH 4-9, 15°C-28°C, 100-200r / min, 5-14d.

[0035] More preferably, the culture conditions include: pH 7, 22°C, 150 r / min, 7 days.

[0036] In a preferred embodiment, the palmate rose ear liquid stock culture medium comprises: 30 g maltose, 10 g peanut powder, 2 g potassium dihydrogen phosphate, 1 g magnesium sulfate, 20 mg vitamin B1, 1000 mL water, pH 7.0.

[0037] Preferably, the method further comprises a multiplication step.

[0038] The propagation comprises: inoculating the palmate Rose fungus strain into a propagation culture medium, inverting and culturing at 22° C. in the dark, terminating the culture when the palmate Rose fungus hyphae cover the surface of the plate culture medium, and storing and setting aside.

[0039] The enrichment medium comprises: 200 g potato, 20 g glucose, 3 g peptone, 2 g potassium dihydrogen phosphate, 1 g magnesium sulfate, 10 mg vitamin B1, 20 g agar, and 1000 mL water.

[0040] In a preferred embodiment, a method for culturing palmate rose fungus using Cordyceps militaris cultivation residues comprises the following steps:

[0041] Step 1: Inoculate the palmate rose ear strain into the propagation medium, invert and culture at 22°C-25°C in the dark, and terminate the culture when the palmate rose ear hyphae cover the surface of the plate medium to obtain the palmate rose ear propagation strain;

[0042] Step 2, inoculating the palmate rose fungus propagation strain into a mother culture medium, inverting and dark-proofing and culturing at 22° C.-25° C., pH 7-9 for 15-20 days to obtain a palmate rose fungus mother culture;

[0043] Step 3, inoculating the palmate rose ear mother seed into the palmate rose ear liquid stock culture medium at pH 4-9, 15° C.-28° C., 100-200 r / min, and culturing at a constant temperature and shaking for 5-14 days to obtain the palmate rose ear liquid stock;

[0044] Step 4: Inoculate the palmate rose fungus liquid stock into the palmate rose fungus cultivation medium at an inoculation volume of 10-15 mL / bag, and perform germ management in a light-proof environment at 18°C-22°C and a relative humidity below 70%;

[0045] Step 5: When the mycelium spreads and grows throughout the culture medium, budding management is performed under the conditions of 300-500 Lux scattered light for 12-16 hours per day, a carbon dioxide concentration of 800-1000 ppm, a humidity of 80%-90%, and 16°C-22°C;

[0046] Step 6: When the light pink palmate rose ear primordium appears, carry out mushroom fruiting management under the conditions of 300-500 Lux diffuse light for 12-16 hours / day, carbon dioxide concentration of 800-1000 ppm, humidity of 80%-90%, and 20℃-22℃.

[0047] On the other hand, the present application also provides the palmate rose fungus cultivation medium or the palmate rose fungus obtained by culturing the palmate rose fungus by the method described; preferably, the palmate rose fungus contains antioxidant active substances; more preferably, the antioxidant active substances include polysaccharides, flavonoids and / or polyphenols.

[0048] On the other hand, the present application also provides the palmate rose fungus cultivation medium or the method or the use of the palmate rose fungus in preparing antioxidant active substances and / or antioxidant; preferably, the antioxidant active substances include polysaccharides, flavonoids and / or polyphenols.

[0049] The present invention has the following beneficial effects:

[0050] 1. This application is the first to apply Cordyceps militaris cultivation residues to the cultivation process of palmate rose fungus, providing a new resource recycling method for Cordyceps militaris cultivation residues, solving the resource waste and environmental pollution problems of Cordyceps militaris cultivation residues, and providing a new low-cost high-quality cultivation resource for palmate rose fungus, while solving the problem of high palmate rose fungus cultivation cost.

[0051] 2. The present application provides a cultivation method for palmate rose fungus, which utilizes the culture residues of Cordyceps militaris to efficiently cultivate palmate rose fungus, thereby realizing the rapid cultivation of palmate rose fungus. The cultivated palmate rose fungus mycelium grows well and has good quality, and can be full of bags in 35 days. The cultivation cycle can be as fast as 47 days. In addition, the palmate rose fungus cultivated by this method also has a high fruiting body yield, with the fruiting body yield being as high as 81.2%. The fruiting body contains a variety of antioxidant active substances, such as total polysaccharides, flavonoids, polyphenols, etc., and has good antioxidant capacity. BRIEF DESCRIPTION OF THE DRAWINGS

[0052] The drawings described herein are used to provide a further understanding of the present application and constitute a part of the present application. The illustrative embodiments of the present application and their descriptions are used to explain the present application and do not constitute an improper limitation on the present application. In the drawings:

[0053] Figure 1 This is a schematic diagram of the growth of palmate Rosy Ear fungus on different culture media in Example 1;

[0054] Figure 2 This is a schematic diagram of the growth of palmate Rosy Ear fungus on different carbon source media;

[0055] Figure 3 This is a schematic diagram of the growth of palmate Rosy Ear fungus on different nitrogen source media;

[0056] Figure 4 The growth of Rosy palmatum strains on different pH culture media;

[0057] Figure 5 This is a schematic diagram of the growth of palmate Roseus mycelium under different culture temperature conditions;

[0058] Figure 6 The figure is a schematic diagram of the growth of the palmate rose fungus strain on the optimized liquid culture medium. Figure A is a microscopic morphology of the hyphae, with a magnification of 40 times, and Figures B and C are sensory morphology of the liquid strain;

[0059] Figure 7 This is a schematic diagram of the mycelium growth and fruiting body growth of the palmate rose fungus experimental group and the control group in Example 4. DETAILED DESCRIPTION

[0060] In order to more clearly explain the overall concept of the present application, the following is described in detail in conjunction with the accompanying drawings of the specification by way of embodiment. In the following description, a large number of specific details are provided to provide a more thorough understanding of the present invention. However, it is obvious to those skilled in the art that the present invention can be implemented without one or more of these details. In other examples, in order to avoid confusion with the present invention, some technical features well known in the art are not described.

[0061] If no specific conditions are specified in the examples, the experiments were carried out according to conventional conditions or conditions recommended by the manufacturer.

[0062] Unless otherwise specified, in the following embodiments, the reagents or instruments used without indicating the manufacturer are all conventional products that can be purchased commercially.

[0063] Unless otherwise stated, the experimental methods, detection methods, and preparation methods disclosed in the present invention all adopt conventional techniques in microbiology, biochemistry, analytical chemistry, cell culture, and related fields conventional in the technical field.

[0064] In addition, the "water" mentioned in the present invention includes any feasible water that can be used in the art, such as deionized water, distilled water, ion exchange water, double distilled water, high-purity water, and purified water.

[0065] In the following examples, unless otherwise specified, % means wt %, i.e. weight percentage.

[0066] Among them, Rhodotus palmatus belongs to the Agaricus family (Agaricaceae) and the genus Rhodotus.

[0067] Cordyceps militaris culture medium formula: wheat grains and nutrient solution are prepared according to a mass ratio of 1:1.6-1.7. The nutrient solution includes: 1.0-2.0g of potassium dihydrogen phosphate, 0.5-1.0g of magnesium sulfate, and 1000mL of water.

[0068] Determination method:

[0069] Polysaccharide content determination method: phenol-sulfuric acid method. According to the determination method of edible fungi polysaccharides, the spectrophotometric method in NY / T1676-2023 Determination of crude polysaccharides in edible fungi was used. The glucose mass concentration was used as the horizontal coordinate and the A490 absorbance value was used as the vertical coordinate. The linear regression equation was obtained as follows: y=504071X+0.0125, R 2 =0.9972.

[0070] Polyphenol content determination method: The polyphenol content was determined by spectrophotometry. Referring to the method of "Determination and analysis of total polyphenol and polyphenol monomer content in different varieties of mulberry leaves" by Deng Zhenhua et al., the mass concentration of gallic acid was used as the horizontal coordinate and the corresponding A750 absorbance value was used as the vertical coordinate. The linear regression equation was y=18.684X-0.0266, R2=0.9985.

[0071] Flavonoid content determination method: Aluminum nitrate colorimetric method was used to determine the total flavonoid content. The flavonoid content was determined by referring to the method of Ge Xinhui et al. on "Analysis of the Composition and Antioxidant Activity of Phenolic Substances in Wild Edible Fungi in Wutai Mountain", with the mass concentration of rutin as the horizontal axis and the corresponding A510 absorbance as the vertical axis, and the linear regression equation was obtained as y=2.9284X+0.0182, R2=0.9974.

[0072] The hydroxyl radical scavenging rate (%) determination method includes: using a hydroxyl radical scavenging ability determination kit for determination and analysis, and the hydroxyl radical scavenging rate of the test sample solution is calculated as follows: hydroxyl radical scavenging rate / % = [A 空白 -(A 测定 -A 对照 )]÷A 空白 ×100.

[0073] The DPPH scavenging ability determination method includes: using a DPPH free radical scavenging ability determination kit for determination, and the DPPH free radical scavenging rate of the test sample solution is calculated by the formula: DPPH free radical scavenging rate / % = [1-(A 测定 -A 对照 )÷A 空白 ]×100; where: A 测定 A is the absorbance of the mixed solution of sample and DPPH; 对照 A is the absorbance of the mixture of sample and anhydrous ethanol; 空白 is the absorbance of the mixture of DPPH and anhydrous ethanol.

[0074] The total antioxidant activity determination method includes: using 2,2'-azobis-3-ethylbenzothiazoline-6-sulfonic acid method, using T-AOC determination kit for determination and analysis, the total antioxidant capacity of the test sample solution is calculated as follows: total antioxidant capacity ( / μmol Trolox / mL) = 0.3 × (△A + 0.029) × D; where: △A = A 空白 -(A 测定 -A 对照 ), where A 测定 is the absorbance of the sample and ABTS working solution, A 对照 A is the absorbance of the mixture of sample and PBS, 空白 is the absorbance of the mixture of ABTS working solution and PBS; D is the dilution multiple of the sample.

[0075] The superoxide anion determination method includes: using a superoxide anion kit for determination, and the superoxide anion content of the test sample solution is calculated as follows: superoxide anion scavenging capacity (μmol / g) = 237.1×

[0076] (△A+0.0011) / W×D; where: △A=A测定 -A 空白 ), where A 测定 is the absorbance of the sample and the color developing solution, A 空白 is the absorbance of the extraction solvent and the color developing solution; W is the sample weight, and D is the dilution multiple of the sample.

[0077] Example 1 Optimization of the culture medium and culture conditions of the palmate rose ear mother seed

[0078] 1. Expansion and cultivation of bacteria

[0079] Plate culture medium configuration: 200 g potato, 20 g glucose, 3 g peptone, 2 g potassium dihydrogen phosphate, 1 g magnesium sulfate, 10 mg vitamin B1, 20 g agar, 1000 mL water; after preparing the culture medium according to the above ratio, sterilize it at 121°C for 25 min, aseptically pour it into a culture dish to prepare the expansion plate culture medium, and set aside;

[0080] Propagation of strains: inoculate the palmate Rosea strain into the center of the propagation plate culture medium, invert and culture at 22°C in the dark. When the palmate Rosea hyphae cover the surface of the plate culture medium, terminate the culture and store it for later use.

[0081] 2. Effects of different culture medium formulas on the growth of Rosa palmata hyphae

[0082] Since the mycelium of palmate rose fungus grows slowly, the components of the mother culture medium were screened in this embodiment. The formula of the mother culture medium is as follows:

[0083] Mother culture medium 1#: glucose 20g, peptone 2g, yeast powder 2g, potassium dihydrogen phosphate 2g, magnesium sulfate 1g, agar 20g, water 1000mL;

[0084] Mother culture medium 2#: glucose 20g, yeast powder 2g, potassium dihydrogen phosphate 2g, magnesium sulfate 1g, vitamin B1 20mg, agar 20g, water 1000mL;

[0085] Mother culture medium 3#: wheat bran 10g, glucose 20g, yeast extract 1.0g, potassium dihydrogen phosphate 2g, magnesium sulfate 1g, vitamin B1 20mg, agar 20g, water 1000mL;

[0086] Mother culture medium 4#: 5g soybean powder, 2.5g peptone, 8g glucose, 2g yeast extract, 1000mL water;

[0087] Mother culture medium 5#: 200g potatoes (boiled in water to extract juice), 30g wheat grains (pre-soaked in boiled water for 20 minutes, filtered to extract juice), 25g glucose, 5g soybean powder, 2g potassium dihydrogen phosphate, 1g magnesium sulfate, 20mg vitamin B1, 20g agar, 1000mL water;

[0088] Mother culture medium 6#: 200g potato, 25g glucose, 5g soybean powder, 5g wheat flour, 2g potassium dihydrogen phosphate, 1g magnesium sulfate, 20mg vitamin B1, 20g agar, 1000mL water;

[0089] Mother culture medium 7#: 200g potato, 25g glucose, 10g soybean powder, 10g peptone, 2g potassium dihydrogen phosphate, 1g magnesium sulfate, 20mg vitamin B1, 20g agar, 1000mL water;

[0090] Mother culture medium 8#: glucose 20g, soybean powder 15g, potassium dihydrogen phosphate 2g, magnesium sulfate 1g, vitamin B1 20mg, agar 20g, water 1000mL;

[0091] Mix the mixture according to the above ratio, sterilize at 121°C for 25 min, and then pour into a plate to obtain the mother culture medium; use a sterile punch to divide the palmate rose fungus strain propagated in step (1) into about 0.3 cm 2 Small pieces of culture medium were aseptically inoculated in the center of different mother culture media, inverted and cultured at 22°C in the dark until the culture medium was covered, and the growth of mycelium was observed and the growth rate of mycelium was measured.

[0092] In this example, the growth of palmate Rosy fungus on different culture media is as follows: Figure 1 And as shown in Table 1.

[0093] Table 1 Effects of different culture media on the growth of Rosa palmata hyphae

[0094]

[0095] Depend on Figure 1 As shown in Table 1, the palmate rose fungus can grow on all 8 culture media, but there are obvious differences in the growth rate and growth potential of the palmate rose fungus mycelium on different culture media. Among them, the mycelium grows fastest on culture medium 8, and the mycelium is dense and vigorous. Therefore, the mother culture medium 8# was selected as the basic culture medium for subsequent experiments.

[0096] 3. Effects of different carbon and nitrogen sources on the growth of Rosa palmata hyphae

[0097] The optimal culture medium formula was obtained by the experiment on the effect of different culture medium formulas on the growth of palmate rose fungus hyphae, which was the mother culture medium 8#, and this culture medium was used as the basic culture medium for subsequent experiments. In this embodiment, a screening test was also conducted on the types of carbon sources and nitrogen sources in the culture medium and the pH value. The details are as follows:

[0098] Basic culture medium: glucose 20g, soybean powder 15g, potassium dihydrogen phosphate 2g, magnesium sulfate 1g, vitamin B1 20mg, agar 20g, water 1000mL;

[0099] Carbon source test: Replace the glucose in the basal medium with the same mass of sucrose, lactose, maltose, and mannitol. Pour the prepared medium into 250 mL Erlenmeyer flasks, 100 mL per flask. Sterilize under high pressure, cool and pour into a flat plate. Use a sterile cork punch to split the palmately shaped Rosella strain propagated in step (1) into approximately 0.3 cm 2 The small pieces were sterilely inoculated in the center of different solid plate culture media, inverted and cultured at 22°C in the dark, and the mycelium growth was observed and the mycelium growth rate was measured. Figure 2 And as shown in Table 2.

[0100] Nitrogen source test: Use cornmeal, malt powder, wheat bran, peanut powder, wheat flour, ammonium sulfate, and ammonium chloride to replace the soybean powder in the basic culture medium. The above-prepared culture medium is respectively placed in 250mL triangular flasks, 100mL per bottle. After high-pressure sterilization and cooling, pour it into a flat plate, and use a sterile punch to divide the palmate rose fungus strain propagated in step (1) into about 0.3cm 2 The small pieces were sterilely inoculated in the center of different solid plate culture media, inverted and cultured at 22°C in the dark, and the mycelium growth was observed and the mycelium growth rate was measured. Figure 3 And as shown in Table 3.

[0101] Table 2 Effect of carbon source on the growth of Rosa palmata hyphae

[0102]

[0103] from Figure 2 As shown in Table 2, the palmate rose fungus can grow on all five carbon source media, and the average daily growth rate of the mycelium is not significantly different, but there are obvious differences in the growth of the mycelium. Among them, the growth rate of the mycelium on the maltose carbon source medium is slightly faster than that on other carbon sources, but the mycelium grows most vigorously and the mycelium is dense. The growth of the mycelium is from strong to weak in the order of maltose>glucose>mannitol>lactose>sucrose. Therefore, the best carbon source for the growth of palmate rose fungus mycelium is maltose.

[0104] Table 3 Effect of nitrogen source on the growth of Rosa palmata hyphae

[0105]

[0106] Depend on Figure 3 As shown in Table 3, the palmate rose fungus can grow on all six organic nitrogen source media. The average daily growth rate of mycelium is not significantly different, but the growth potential is significantly different. No growth was observed on the media with ammonium sulfate and ammonium chloride as nitrogen sources. The average daily growth rate of mycelium on the peanut cake powder medium was the fastest and the growth potential was the best. Therefore, the best nitrogen source for the growth of palmate rose fungus mycelium is peanut cake powder.

[0107] 4. Effects of different pH values ​​on the growth of Rosa palmata mycelium

[0108] Use 1.0N HCl and 1.0N NaOH to adjust the pH of the basal medium to 4, 5, 6, 7, 8, and 9, respectively. Pour the prepared medium into 250mL Erlenmeyer flasks, 100mL per flask. After autoclaving and cooling, pour the plate and use a sterile cork punch to split the palmately shaped Rosella strain propagated in step (1) into approximately 0.3cm 2 The small pieces were sterilely inoculated in the center of different solid plate culture media, inverted and cultured at 22°C in the dark, and the mycelium growth was observed and the mycelium growth rate was measured. Figure 4 And as shown in Table 4.

[0109] Table 4 Effect of pH on the growth of Rosa palmata hyphae

[0110]

[0111] Depend on Figure 4 As shown in Table 4, the palmate rose fungus can grow on all six pH culture media, and the average daily growth rate and growth potential of the mycelium are significantly different. In terms of growth rate, as the pH increases, the average daily growth rate of the mycelium gradually increases. When the pH of the culture medium is 7.0-9.0, the mycelium growth rate is comparable and reaches a higher level. The mycelium growth is optimal at pH 7.0. Therefore, pH 7.0 is selected as the optimal pH for the growth of palmate rose fungus mycelium.

[0112] Based on the comprehensive test results, the formula of the culture medium for the palmate rose mother seed was determined to be: 20g maltose, 15g peanut cake powder, 2g potassium dihydrogen phosphate, 1g magnesium sulfate, 20mg vitamin B1, 20g agar, 1000mL water, pH 7.0.

[0113] 5. Effects of different temperatures on the growth of P. palmatum mycelium

[0114] In this example, the effects of different temperature conditions on the growth of Rosa palmata mycelium were also investigated.

[0115] Use a sterile punch to split the palmately shaped Rosella strain propagated in step (1) into approximately 0.3 cm 2 The small pieces were aseptically inoculated in the center of the solid plate culture medium optimized in step (4), inverted and cultured at 16°C-30°C in the dark, and the mycelial growth was observed and the mycelial growth rate was measured. Figure 5 And as shown in Table 5.

[0116] Table 5 Effect of temperature on the growth of Rosa palmata hyphae

[0117]

[0118] Depend on Figure 5 As shown in Table 5, temperature has a significant effect on the growth and growth rate of palmate rose mycelium. In terms of growth rate, with the increase of culture temperature, the growth rate of palmate rose mycelium increases first and then decreases. When the culture temperature reaches 30℃, the mycelium stops growing. The growth rate is fastest at 22℃ and 25℃, and the daily growth rate of mycelium reaches 0.358cm / d and 0.352cm / d respectively. In terms of mycelium growth, the palmate rose mycelium can grow well in the culture temperature range of 16℃-28℃. The palmate rose mycelium is more vigorous and the colony is dense. Considering the mycelium growth rate, the best culture temperature is 22℃-25℃.

[0119] Example 2 Optimization of the liquid stock culture medium formula of palmate rose fungus

[0120] 1. The conditions optimized in Example 1 were used to carry out the propagation and mother culture of the palmately shaped rose fungus.

[0121] 2. Preparation of different palmate rose fungus liquid stock culture medium:

[0122] According to the test results of Example 1, the liquid stock culture medium formula of the palmate rose fungus was optimized, and the effects of different added amounts of maltose and peanut cake powder on the liquid culture of mycelium were investigated using the optimized mother culture medium formula as the liquid basal culture medium.

[0123] Liquid basal culture medium: 20 g maltose, 15 g peanut powder, 2 g potassium dihydrogen phosphate, 1 g magnesium sulfate, 20 mg vitamin B1, pH 7.0, 1000 mL water.

[0124] Maltose addition test: 10g, 20g, 30g, and 40g of maltose were added to each liter of liquid basal culture medium to prepare the palmate rose ear liquid stock culture medium.

[0125] Peanut cake powder addition amount test: Based on the test of the optimal addition amount of maltose, a peanut cake powder addition amount test was carried out. 5g, 10g, 15g and 20g of peanut cake powder were added to each liter of liquid basic culture medium to prepare palmate rose ear liquid original culture medium.

[0126] After preparing the culture medium according to the above ratio, dispense it into 250 mL Erlenmeyer flasks, with 100 mL in each bottle. Sterilize at 121°C for 25 min and set aside.

[0127] 3. Cultivation of palmate rose fungus:

[0128] Five pieces of each palmate Rosa odorata strain were inoculated into different culture media, and cultured at 22°C and 150 r / min for 7 days with constant temperature shaking. 100 mL of the fermentation broth was taken into a 100 mL measuring cylinder and allowed to stand for 30 min to examine the volume ratio of mycelium in the fermentation broth in different culture media.

[0129] The effect of the amount of maltose added on the mycelial biomass in this example is shown in Table 6, and the effect of the amount of peanut cake powder added on the mycelial biomass is shown in Table 7.

[0130] Table 6 Effect of maltose addition on the mycelial growth of Rosa digitata liquid culture

[0131] Maltose addition amount / (g) 10 20 30 40 Mycelium volume ratio in fermentation broth / (%) 24.5 28.6 40.3 39.6

[0132] It can be seen from Table 6 that different concentrations of maltose in the liquid stock culture medium of Palmate Rose Fungus have a significant effect on the mycelial growth. With the increase of maltose addition, the amount of mycelium gradually increases. When the addition amount is 30g, the amount of mycelium reaches the maximum. Continuing to increase the amount of mycelium does not increase. Therefore, the optimal addition amount of maltose is 30g.

[0133] Table 7 Effect of peanut powder addition on the mycelial growth of Rosa digitata liquid culture

[0134] Amount of peanut powder added / (g) 5 10 15 20 Mycelium volume ratio in fermentation broth / (%) 35.3 41.5 28.6 28.5

[0135] It can be seen from Table 7 that different concentrations of peanut cake powder in the liquid stock culture medium of Palmate Rose Ear have a significant effect on the growth of mycelium. With the increase of the amount of peanut cake powder added, the amount of mycelium shows a trend of gradually increasing and then decreasing. When the added amount is 10g, the amount of mycelium reaches the maximum. Continuing to increase the amount of mycelium does not increase but decreases. Therefore, the optimal amount of peanut cake powder added is 10g.

[0136] According to the test results of maltose and peanut powder addition, the best liquid culture medium formula of palmate rose ear is determined as follows: 30g maltose, 10g peanut powder, 2g potassium dihydrogen phosphate, 1g magnesium sulfate, 20mg vitamin B1, pH 7.0, and 1000mL water. Figure 6 shown.

[0137] Depend on Figure 6 It can be seen that the ball of palmate rose fungus is dense on the optimized palmate rose fungus liquid stock culture medium, and the mycelium volume accounts for 42.9%, which is significantly higher than 28.6% before optimization. Therefore, the optimal palmate rose fungus liquid stock culture medium formula is: 30g maltose, 10g peanut cake powder, 2g potassium dihydrogen phosphate, 1g magnesium sulfate, 20mg vitamin B1, pH 7.0, and 1000mL water.

[0138] Example 3 Effects of different addition amounts of Cordyceps militaris cultivation residues on the growth of palmate rose fungus fruiting bodies

[0139] 1. Preparation of palmate rosehip seeds

[0140] (1) The palmate rose mother culture medium is the solid slant culture medium selected in Example 1, and its formula is: 20 g maltose, 10 g peanut powder, 2 g potassium dihydrogen phosphate, 1 g magnesium sulfate, 20 mg vitamin B1, pH 7.0, 15 g agar, and 1000 mL water.

[0141] (2) Aseptically inoculate fresh culture of the Rosa palmatum strain into the mother culture medium, invert and culture at 22°C in the dark for 15-20 days until it grows to cover the entire medium.

[0142] 2. Preparation of Liquid Stock of Palmate Rose Fungus

[0143] (1) The liquid culture medium for palmate rose ear is the liquid culture medium selected in Example 2, and its formula is: 30 g maltose, 10 g peanut powder, 2 g potassium dihydrogen phosphate, 1 g magnesium sulfate, 20 mg vitamin B1, pH 7.0, and 1000 mL water.

[0144] (2) The culture method of liquid stock is the same as that in Example 2.

[0145] 3. Preparation of culture medium

[0146] (1) Cultivation medium formula

[0147] Using Cordyceps militaris cultivation residues as the main raw material, five cultivation medium formulas were designed as follows:

[0148] Cultivation medium 1#: 800g sawdust, 200g Cordyceps militaris cultivation residue, 10g quicklime, and the balance is water, with a water content of 55%-60%;

[0149] Cultivation medium 2#: 700g sawdust, 300g Cordyceps militaris cultivation residue, 10g quicklime, add water to make the water content in the culture medium reach 55%-60%;

[0150] Cultivation medium 3#: 600g sawdust, 400g Cordyceps militaris cultivation residue, 10g quicklime, add water to make the water content in the culture medium reach 55%-60%;

[0151] Cultivation medium 4#: 500g sawdust, 500g Cordyceps militaris cultivation residue, 10g quicklime, add water to make the water content in the culture medium reach 55%-60%;

[0152] Cultivation medium 5#: 400g sawdust, 600g Cordyceps militaris cultivation residue, 10g quicklime, and water is added to make the water content in the culture medium reach 55%-60%.

[0153] (2) Method of preparing cultivation materials

[0154] The Cordyceps militaris cultivation residue needs to be soaked in water for 3-4 hours before use, drained, mixed with other materials according to the ratio and stirred evenly, and then water is added and stirred to make the moisture in the culture medium reach 55%-60%, and then put into a polypropylene bag with a specification of 17cm×33cm, a material height of 10cm, a loading amount of 260g (dry material), sealed, and sterilized at 121℃-125℃ for 2 hours.

[0155] 4. Inoculation

[0156] Place the sterilized polypropylene cultivation bag in a sterile environment. When the material temperature drops below 22°C, put it into the disinfected inoculation box. Inoculate the cultured palmate rosea liquid stock from the sealed area according to the sterile operating procedures. Inoculate each cultivation bag with 10 mL of liquid stock. Seal the bag after inoculation.

[0157] 5. Bacteria management

[0158] Wipe the culture room and culture rack with disinfectant and sterilize. Transfer the inoculated polypropylene bag to the culture rack for spawn culture. Ventilate every day during the entire spawn culture period. Control the room temperature at a constant temperature of 18℃-22℃ and the relative humidity of the air is lower than 70%. Cultivate in the dark and observe the growth of mycelium. When the mycelium spreads and grows to the entire culture medium, you can start mushroom production management.

[0159] 6. Mushroom production management

[0160] Transfer the polypropylene bags after the spawning to the fruiting room, place them upright, remove the plugs of the bags, open the bags, and manage the budding and fruiting under the conditions of 300-500 Lux scattered light for 12-16 hours / day, carbon dioxide concentration of 800-1000ppm, and humidity of 80%-90%. Among them, the culture temperature is controlled at 16℃-22℃ during the budding stage. When the light pink palmate rose ear primordium appears, adjust the culture temperature to 20℃-22℃ for fruiting management. When the fruiting body cap is unfolded, harvest the fruiting body in time, harvest one crop from each bag, and examine the fruiting body yield.

[0161] The mycelial growth and bag-filling time during the culture process were monitored and the fruiting body yield was recorded. The specific results are shown in Table 8.

[0162] Table 8 Mycelial growth on different cultivation media

[0163] formula Mycelium growth Full bag time (days) Fruiting body yield (%) Cultivation medium 1# Mycelium is white and vigorous 40 50.4 Cultivation medium 2# Mycelium is white and vigorous 38 55.8 Cultivation medium 3# Mycelium is white and vigorous 35 71.5 Cultivation medium 4# Mycelium is white and vigorous 35 80.8 Cultivation medium 5# Mycelium is white and vigorous 37 61.2

[0164] As shown in Table 8, the mycelium and fruiting body of the palmate rosy fungus can grow well on the culture medium containing 200-600 g of the Cordyceps militaris cultivation residues, which proves that the Cordyceps militaris cultivation residues can be used to cultivate the palmate rosy fungus.

[0165] Among them, the content of Cordyceps militaris cultivation residues is in the range of 200-500g. With the increase of the added amount, the mycelium and the speed of eating the material, and the fruiting body yield are significantly improved, and then with the increase of the added ratio, the mycelium growth rate and the fruiting body yield decrease; Among them, adding 500g of Cordyceps militaris cultivation residues to the cultivation medium 4# has the best effect. At this time, the shortest time for the mycelium to eat the cultivation material is 35 days, the fruiting body yield reaches more than 80.8%, and the cultivation effect is the best.

[0166] Example 4 Cultivation effect and activity evaluation of palmate rose fungus under optimized conditions

[0167] 1. Prepare the palmate rose ear mother seed and liquid stock seed according to the method and culture medium of Example 3.

[0168] 2. Cultivate palmate rose fungus according to the cultivation medium optimized according to implementation case 3.

[0169] 3. Preparation of culture medium:

[0170] (1) Cultivation medium formula: 500 g sawdust, 500 g Cordyceps militaris cultivation residue, 10 g quicklime, and water is added to make the water content in the culture medium reach 55%-60%;

[0171] In this embodiment, a culture medium without adding Cordyceps militaris cultivation residues is used as a control cultivation medium. The control cultivation medium formula is: 680g sawdust, 100g cottonseed hulls, 150g wheat bran, 50g soybean meal, 10g gypsum, and 10g lime. Water is added to make the moisture content in the culture material reach 55%-60%.

[0172] (2) The method of preparing the cultivation material is the same as that in Example 3.

[0173] 4. Inoculation is the same as in Example 3.

[0174] 5. The germ management is the same as in Example 3.

[0175] 6. The mushroom production management is the same as in Example 3.

[0176] In this example, the growth and active substance content of palmate rose fungus were measured, and the results are shown in Table 9. Figure 7 , as shown in Table 10 and Table 11.

[0177] Table 9 Example 5 Palmate Rose Ear Growth

[0178] Mycelium growth Full bag time Primordial formation time Cultivation cycle Fruiting body yield% Optimized culture medium Strong and vigorous 35 days 38-39 days 47 days 81.2 Control culture medium Strong and vigorous 50 days 54-57 65 48.7

[0179] Table 10 Active substance content in the fruiting bodies of palmate rose ears in Example 5 and the fruiting bodies of the control group

[0180]

[0181] Table 11 Antioxidant activity of active substances in palmate rose fungus fruiting bodies cultured on optimized cultivation medium in Example 5

[0182]

[0183] from Figure 7 As shown in Table 9, under the optimized conditions, the culture matrix containing 50% of the Cordyceps militaris cultivation residues provided good growth conditions for the palmate fungus. The palmate fungus mycelium grew vigorously and densely, and the mushroom bag was fully grown in 35 days. At 15℃-22℃, light pink palmate fungus primordia were formed in 3-4 days. The fruiting bodies matured after continued cultivation in the range of 20-22℃ for 7-8 days. The yield of the single-flow mushroom fruiting bodies reached 81.2%.

[0184] This technology not only has a simple and easy-to-configure cultivation medium, but also innovates the palmate rose fungus cultivation matrix, replacing 50% of sawdust, bran and other raw materials by mass with Cordyceps militaris cultivation residues, saving forestry resources, reducing the cost of cultivation raw materials by more than 13%, achieving a one-time burst of mushroom production, and a single-wave mushroom fruiting body yield of more than 80%, significantly improving the cultivation efficiency of palmate rose fungus and making it easier for factory production. At the same time, it realizes the large-scale secondary utilization of Cordyceps militaris cultivation residues, turning waste into treasure, reducing resource waste and environmental pollution, and is of great significance to the efficient, green and sustainable development of the rare edible fungi industry.

[0185] Further, it can be seen from Table 10 that, compared with the control group, the fruiting body yield of the palmate rose ear cultivated by replacing bran, sawdust and other components in the culture medium with the cultivation residues of Cordyceps militaris was increased from 48.7% to 81.2%, and the contents of total polysaccharides, polyphenols and flavonoids in the fruiting body were increased by 1.39, 1.33 and 1.02 times, respectively. This is mainly because the cultivation residues of Cordyceps militaris are discarded wheat grains after the fruiting bodies of Cordyceps militaris are harvested, which increases the air permeability of the culture medium. At the same time, the cultivation residues obtained after the cultivation of Cordyceps militaris are rich in nutritional factors such as starch, protein, and cordyceps phenolic compounds, which provide a rich nutritional environment for the growth of palmate rose ear fruiting bodies and promote the growth of the fruiting bodies and the synthesis of polysaccharides and flavonoid compounds.

[0186] It can be further seen from Table 11 that the palmate rose fungus fruiting body has good antioxidant activity and is a rare renewable resource for antioxidant foods and health products.

[0187] The above is only an embodiment of the present application and is not intended to limit the present application. For those skilled in the art, the present application may have various changes and variations. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present application should be included in the scope of the claims of the present application.

Claims

1. Application of Cordyceps militaris cultivation residues in cultivating palmate rose fungus.

2. The use according to claim 1, characterized in that: The Cordyceps militaris cultivation residue is obtained by culturing Cordyceps militaris in a Cordyceps militaris culture medium; preferably, the Cordyceps militaris culture medium comprises wheat grains, potassium dihydrogen phosphate and / or magnesium sulfate.

3. A palmate rose ear cultivation medium, characterized in that: The palmate rose fungus cultivation medium comprises Cordyceps militaris cultivation residues.

4. The palmate rose ear cultivation medium according to claim 3, characterized in that: The Cordyceps militaris cultivation residue is obtained by culturing Cordyceps militaris in a Cordyceps militaris culture medium; preferably, the Cordyceps militaris culture medium comprises wheat grains, potassium dihydrogen phosphate and / or magnesium sulfate.

5. The palmate rose ear cultivation medium according to claim 3, characterized in that: Calculated by the mass fraction of the culture medium, the content of the Cordyceps militaris cultivation residue is 200-600 parts; preferably, 200-500 parts; more preferably, 500 parts.

6. The palmate rose ear cultivation medium according to claim 3, characterized in that: The culture medium further comprises sawdust and / or quicklime; preferably, 400-800 parts of sawdust; preferably, 8-15 parts of quicklime; and / or, The culture medium also includes water; preferably, the water content of the culture medium is 55%-60% based on the mass percentage of the culture medium.

7. A method for cultivating palmate rose fungus using Cordyceps militaris cultivation residues, characterized in that: The method comprises: cultivating palmate rose fungus using the palmate rose fungus cultivation medium as described in any one of claims 3-6.

8. The method according to claim 7, characterized in that The method comprises the following steps: Step 1: Inoculate the palmate rose fungus into the palmate rose fungus cultivation medium at an inoculation volume of 10-15 mL / bag, and perform germ management in a light-proof environment at 18°C-22°C and a relative humidity of less than 70%; Step 2: Perform bud-inducing management under the conditions of 300-500 Lux scattered light for 12-16 hours per day, carbon dioxide concentration of 800-1000 ppm, humidity of 80%-90%, and 16°C-22°C; Step 3: Carry out mushroom production management under the conditions of 300-500 Lux diffuse light for 12-16 hours per day, carbon dioxide concentration of 800-1000 ppm, humidity of 80%-90%, and 20°C-22°C; Preferably, the method further comprises the step of inoculating palmate rose ear into palmate rose ear mother seed culture medium to obtain palmate rose ear mother seed; more preferably, the palmate rose ear mother seed culture medium comprises: a carbon source and / or a nitrogen source; more preferably, the carbon source is selected from one or more of sucrose, lactose, glucose, maltose, and mannitol; more preferably, the carbon source is maltose; more preferably, the nitrogen source is selected from one or more of bean powder, cornmeal, malt powder, wheat bran, ammonium sulfate, ammonium chloride, peanut cake powder, and wheat grain powder; more preferably, the nitrogen source is peanut cake powder; more preferably, the palmate rose ear mother seed culture medium further comprises: potassium dihydrogen phosphate, magnesium sulfate and / or vitamin B1; more preferably, the palmate rose ear mother seed culture medium comprises: 1%-5% maltose, 1%-5% peanut cake powder, 0.1%-0.5% potassium dihydrogen phosphate, 0.1%-0.5% magnesium sulfate, vitamin B1 0.001%-0.005%; more preferably, the palmate rose mother culture medium includes: 2% maltose, 1.5% peanut powder, 0.2% potassium dihydrogen phosphate, 0.1% magnesium sulfate, and 0.002% vitamin B1; preferably, the culture conditions include: 16°C-28°C, pH4-9; more preferably, 22°C-25°C, pH7-9, 15-20d; Preferably, the method further comprises the step of inoculating the palmate rose ear mother seed into the palmate rose ear liquid stock culture medium to obtain the palmate rose ear liquid stock; more preferably, the palmate rose ear liquid stock culture medium comprises: a carbon source and / or a nitrogen source; more preferably, the carbon source is selected from one or more of sucrose, lactose, glucose, maltose, and mannitol; more preferably, the carbon source is maltose; more preferably, the nitrogen source is selected from one or more of bean powder, cornmeal, malt powder, wheat bran, ammonium sulfate, ammonium chloride, peanut cake powder, and wheat grain powder; more preferably, the nitrogen source is peanut cake powder; more preferably, the palmate rose ear liquid stock culture medium further comprises: potassium dihydrogen phosphate, magnesium sulfate and / or vitamin B1; more preferably, the palmate rose ear liquid stock culture medium comprises: 1%-5% maltose, 1%-5% peanut cake powder, 0.1%-0.5% potassium dihydrogen phosphate, 0.1%-0.5% magnesium sulfate, vitamin B1 0.001%-0.005%; more preferably, the palmate rose ear liquid stock culture medium includes: 3% maltose, 1% peanut powder, 0.2% potassium dihydrogen phosphate, 0.1% magnesium sulfate, and 0.002% vitamin B1; preferably, the culture conditions include: pH 4-9, 15°C-28°C, 100-200r / min, 5-14d.

9. A palmate rose fungus obtained by culturing the palmate rose fungus according to any one of claims 3 to 6 or the method according to claim 7 or 8; preferably, the palmate rose fungus contains antioxidant active substances; more preferably, the antioxidant active substances include polysaccharides, flavonoids and / or polyphenols.

10. Use of the palmate rose fungus cultivation medium as claimed in any one of claims 3 to 6, the method as claimed in claim 7 or 8, or the palmate rose fungus as claimed in claim 9 in the preparation of antioxidant active substances and / or anti-oxidation; preferably, the antioxidant active substances include polysaccharides, flavonoids and / or polyphenols.

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