CN113330984A - Artificial cultivation method for improving cordyceps sobolifera polysaccharide content in cordyceps sobolifera sporocarp - Google Patents
Artificial cultivation method for improving cordyceps sobolifera polysaccharide content in cordyceps sobolifera sporocarp Download PDFInfo
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- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 claims description 8
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- ZPWVASYFFYYZEW-UHFFFAOYSA-L dipotassium hydrogen phosphate Chemical compound [K+].[K+].OP([O-])([O-])=O ZPWVASYFFYYZEW-UHFFFAOYSA-L 0.000 claims description 5
- 235000019341 magnesium sulphate Nutrition 0.000 claims description 5
- 235000019796 monopotassium phosphate Nutrition 0.000 claims description 5
- 235000019319 peptone Nutrition 0.000 claims description 5
- PJNZPQUBCPKICU-UHFFFAOYSA-N phosphoric acid;potassium Chemical compound [K].OP(O)(O)=O PJNZPQUBCPKICU-UHFFFAOYSA-N 0.000 claims description 5
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- 235000003283 Pachira macrocarpa Nutrition 0.000 claims description 3
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Classifications
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G18/00—Cultivation of mushrooms
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G18/00—Cultivation of mushrooms
- A01G18/20—Culture media, e.g. compost
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- Life Sciences & Earth Sciences (AREA)
- Mycology (AREA)
- Environmental Sciences (AREA)
- Mushroom Cultivation (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The invention belongs to the technical field of artificial cultivation of edible fungi, and particularly relates to an artificial cultivation method for improving the polysaccharide content in cordyceps sobolifera sporocarp. According to the cultivation method for improving cordyceps sobolifera polysaccharide in cordyceps sobolifera sporocarp, on the basis of a conventional cordyceps sobolifera sporocarp cultivation process, dead leaves and trimmed overgrown leaves generated in succulent plants are utilized and added into a solid culture medium of cordyceps sobolifera, and surprisingly, the added dead leaves and leaves of the succulent plants can generate effective induction on accumulation of cordyceps sobolifera polysaccharide in the solid fermentation process of cordyceps sobolifera, so that the content of cordyceps sobolifera polysaccharide in the cordyceps sobolifera sporocarp is improved.
Description
Technical Field
The invention belongs to the technical field of artificial cultivation of edible fungi, and particularly relates to an artificial cultivation method for improving cordyceps sobolifera polysaccharide content in cordyceps sobolifera fruiting bodies.
Background
Cordyceps cicadae is one of traditional Chinese medicinal materials, also called golden cordyceps cicadae and cordyceps cicadae, and is a dry complex of fungus paecilomyces cicadae parasitizing on cicada nymphs in Cordyceps of Clavicepitaceae. The existing research results show that the cicada fungus is similar to the main ingredient amino acid in various cordyceps sinensis in type and has different degrees of tonifying effects. The biological active substances contained in cordyceps sobolifera reported at present comprise nucleosides, proteins, saccharides, alkaloids, sterols and the like, and polysaccharides are important substances. Researches find that the cordyceps sobolifera polysaccharide has the pharmacological effects of resisting tumors, inhibiting immunity, enhancing immunity, resisting oxidation and aging, protecting liver, reducing blood pressure and the like.
Because the polysaccharide is often a high polymer formed by connecting a plurality of same monosaccharide groups by glycosidic bonds, the molecular structure of the polysaccharide is complex, the artificial synthesis is difficult, and the polysaccharide is often extracted and prepared from natural products. However, the content of cordyceps cicadae miq extracted from cordyceps cicadae miq is low at present, mainly because the content of cordyceps cicadae miq polysaccharide in artificially cultivated cordyceps cicadae miq is not ideal. Therefore, a cultivation method capable of effectively increasing the content of cordyceps sobolifera polysaccharide in cordyceps sobolifera is urgently needed to be developed.
Succulent plants are higher plants with large vegetative organs, usually with three vegetative organs, root, stem and leaf, and three reproductive organs, flower, fruit and seed. Succulent plants are popular as potted plants around the world, becoming the most popular plants among young people and going to thousands of households. During the cultivation of succulent plants, a certain amount of dead leaves are generated along with the metabolism of the plants, and the leaves are trimmed due to excessive growth. These dead leaves or leaves, even though not large in individual plants, still cause handling troubles for users who grow succulent plants in large quantities.
Disclosure of Invention
Therefore, the technical problem to be solved by the invention is to provide an artificial cultivation method for improving the content of cordyceps sobolifera polysaccharide in cordyceps sobolifera sporocarp, so as to solve the problem that the content of cordyceps sobolifera polysaccharide in cordyceps sobolifera in the prior art is not ideal;
the second technical problem to be solved by the invention is to provide cordyceps sobolifera with higher cordyceps sobolifera polysaccharide content.
In order to solve the technical problems, the artificial cultivation method for improving the content of cordyceps sobolifera polysaccharide in cordyceps sobolifera sporocarp comprises the steps of performing liquid strain cultivation in a liquid culture medium and performing sporocarp cultivation in a solid culture medium;
the solid culture medium comprises the following components in a mass ratio of 1: 0.8-1.5 of solid matrix and nutrient solution;
the solid matrix comprises the following components in parts by weight: 10-20 parts of rice, 20-30 parts of corn grit, 10-20 parts of bran, 10-20 parts of cicada pupa powder, 5-12 parts of withered leaves of an Ji hazy moon plant, 5-12 parts of broken leaves of the Ji hazy moon plant and 0.5-2 parts of EM (effective microorganism) powder;
the nutrient solution comprises the following components in percentage by mass: 20-30g/L of carbon source, 8-15g/L of nitrogen source, 3-8/L of inorganic salt and natural pH.
Wherein, the dull leaf of dimly moon plant is the dry leaf under pruning among the succulent farming process through naturally drying can, and dimly moon plant leaf particle then adopts the succulent farming in-process because the blade under the plant is spindly and pruned, through naturally drying and smash to graininess can.
Specifically, the artificial cultivation method for improving the content of cordyceps sobolifera polysaccharide in cordyceps sobolifera sporocarp comprises the following steps:
the carbon source comprises glucose;
the nitrogen source comprises peptone;
the inorganic salt comprises a mixture of potassium dihydrogen phosphate, dipotassium hydrogen phosphate and magnesium sulfate.
Specifically, the sporocarp culturing step comprises a spawn running step, a color conversion step and a sporocarp management step.
Specifically, the control conditions of the spawn running step comprise: culturing at 15-18 deg.C and humidity of 60-80% under dark condition.
Specifically, the control conditions of the color conversion step include: controlling the illumination condition to be 200-500lux, and controlling the photoperiod light-dark ratio to be L15-18: d6-9, the culture temperature is 15-18 ℃, and the humidity is 60-80%.
Specifically, the control conditions of the step of managing a fruit body include: controlling the illumination condition to be 200-500lux, and controlling the photoperiod light-dark ratio to be L15-18: d6-9, the culture temperature is 20-25 ℃, and the humidity is 60-80%.
Specifically, the liquid culture medium comprises the following components in percentage by mass: 1-2% of glucose, 0.2-0.4% of yeast extract, 0.8-1.2% of water chestnut, 0.5-1% of egg white and MgSO 44 0.01-0.03％、KH2PO40.01-0.03%, natural pH.
Specifically, the conditions for culturing the liquid strain comprise: the fermentation temperature is controlled to be 20-25 ℃, and the stirring speed is 150 and 180 rpm.
Specifically, the artificial cultivation method for improving the content of cordyceps sobolifera polysaccharide in cordyceps sobolifera fruiting bodies further comprises the step of carrying out conventional activation on the strain before the step of culturing the liquid strain.
The invention also discloses cordyceps sobolifera sporocarp obtained by the cultivation method.
According to the method for improving cordyceps sobolifera polysaccharide in cordyceps sobolifera sporocarp, on the basis of the conventional cordyceps sobolifera sporocarp cultivation process, dead leaves and trimmed overgrown leaves generated in succulent plants are utilized and added into a solid culture medium of cordyceps sobolifera, and surprisingly, the added dead leaves and leaves of succulent plants are compounded with the optimally screened solid medium and nutrient solution in the solid fermentation process of cordyceps sobolifera, so that effective induction can be generated on accumulation of cordyceps sobolifera polysaccharide, the content of cordyceps sobolifera polysaccharide in cordyceps sobolifera sporocarp is improved, the nutrient value of cordyceps sobolifera is improved, and support is provided for deep research and utilization of cordyceps sobolifera polysaccharide.
Detailed Description
In the following embodiments of the invention, the selected cordyceps sobolifera-cultivating strain is a conventionally known strain in the prior art, and the cordyceps sobolifera-cultivating strain selected in the following embodiments is purchased from Shanghai Hao Sheng Kogyo Co.
In the following embodiments of the present invention, the cordyceps sobolifera is subjected to liquid culture to obtain a corresponding culture solution (seed solution), and then inoculated into a corresponding solid culture medium to perform fermentation of fruiting bodies. In the following examples, the liquid medium used in the liquid culture step includes the following components by mass: according to the weight percentage of 1.5 percent of glucose, 0.3 percent of yeast extract, 1 percent of water chestnut, 0.8 percent of egg white and MgSO4 0.02％、KH2PO4Taking the components in a proportion of 0.02 percent; adding water into selected amount of corm Eleocharitis, pulping, adding selected amount of glucose, yeast extract, ovum gallus Domesticus album, and MgSO4、KH2PO4Mixing, sealing, sterilizing at 121 deg.C for 20min, taking out culture medium, and naturally cooling to room temperature. Under the aseptic condition, digging blocks of the preserved strains and inoculating the blocks into the liquid culture medium, controlling the fermentation temperature to be 22 ℃, and carrying out shake flask culture at the stirring speed of 160rpm for 48 hours to obtain seed liquid to be inoculated for carrying out solid fermentation on fruit bodies in the following examples for later use.
In the following embodiments of the present invention, the dead leaves of the hazy moon plant are obtained by naturally drying the dead leaves trimmed in the succulent cultivation process, and the crushed particles of the leaves of the hazy moon plant are obtained by naturally drying the trimmed leaves due to overgrowth of the plant in the succulent cultivation process and crushing the crushed particles into particles (preferably, particle size of 1 to 3 mm).
Example 1
In this example, the seed liquid obtained as described above was cultured in a solid medium to obtain a fruit body.
The solid matrix was formulated at the following levels: 10 parts of rice, 30 parts of corn grit, 10 parts of bran, 20 parts of cicada pupa powder, 5 parts of withered leaves of the hazy moon plant, 12 parts of broken leaves of the hazy moon plant and 0.5 part of EM (not added temporarily);
the nutrient solution is prepared according to the following contents: 20g/L glucose, 30g/L peptone, 2g/L potassium dihydrogen phosphate, 1g/L dipotassium hydrogen phosphate and 2g/L magnesium sulfate;
according to the following steps of 1: 0.8, mixing the solid culture medium (except EM bacteria powder) and the nutrient solution uniformly, sealing the prepared culture medium, placing the culture medium in a high-pressure moist heat sterilization pot, and performing conventional sterilization at 121 ℃ for 40 min.
Inoculating the seed solution into the solid culture medium according to the inoculation amount of 10 wt% under the room temperature and aseptic condition, adding the EM bacterial powder with corresponding amount, uniformly mixing, then placing the inoculated culture medium into a cultivation room for cultivation, controlling the temperature at 17 ℃ and the humidity at 70%, and performing light-tight cultivation for 3-5 days until the spawn running is complete.
And then adjusting the illumination condition, controlling the illumination condition to be 300lux, and controlling the photoperiod light-dark ratio to be L16: d8, continuously controlling the culture temperature to be 17 ℃ and the humidity to be 70%, continuously culturing for 5-8 days to finish color conversion and form the stroma bud.
And then maintaining the illumination condition, continuously controlling the illumination condition to be 300lux, and controlling the photoperiod light-dark ratio to be L16: d8, controlling the culture temperature at 22 deg.C and humidity at 70%, continuously culturing for 15-20 days, collecting mature fruiting body, and oven drying.
Example 2
In this example, the seed liquid obtained as described above was cultured in a solid medium to obtain a fruit body.
The solid matrix was formulated at the following levels: 20 parts of rice, 20 parts of corn grit, 20 parts of bran, 10 parts of cicada pupa powder, 12 parts of dead leaves of the hazy moon plant, 5 parts of broken leaves of the hazy moon plant and 2 parts of EM (temporarily not added);
the nutrient solution is prepared according to the following contents: 30g/L glucose, 20g/L peptone, 4g/L potassium dihydrogen phosphate, 4g/L dipotassium hydrogen phosphate and 4g/L magnesium sulfate;
according to the following steps of 1: 1.5, uniformly mixing the solid culture medium (except EM bacteria powder) and the nutrient solution, sealing the prepared culture medium, placing the culture medium in a high-pressure moist heat sterilization pot, and conventionally sterilizing for 40min at 121 ℃.
Inoculating the seed solution into the solid culture medium according to the inoculation amount of 10 wt% under the room temperature and aseptic condition, adding the EM bacterial powder with corresponding amount, uniformly mixing, then placing the inoculated culture medium into a cultivation room for cultivation, controlling the temperature at 17 ℃ and the humidity at 70%, and performing light-tight cultivation for 3-5 days until the spawn running is complete.
And then adjusting the illumination condition, controlling the illumination condition to be 300lux, and controlling the photoperiod light-dark ratio to be L16: d8, continuously controlling the culture temperature to be 17 ℃ and the humidity to be 70%, continuously culturing for 5-8 days to finish color conversion and form the stroma bud.
And then maintaining the illumination condition, continuously controlling the illumination condition to be 300lux, and controlling the photoperiod light-dark ratio to be L16: d8, controlling the culture temperature at 22 deg.C and humidity at 70%, continuously culturing for 15-20 days, collecting mature fruiting body, and oven drying.
Example 3
In this example, the seed liquid obtained as described above was cultured in a solid medium to obtain a fruit body.
The solid matrix was formulated at the following levels: 15 parts of rice, 25 parts of corn grit, 15 parts of bran, 15 parts of cicada pupa powder, 8 parts of dead leaves of the hazy moon plant, 10 parts of broken leaves of the hazy moon plant and 1 part of EM (temporarily not added);
the nutrient solution is prepared according to the following contents: 25g/L glucose, 25g/L peptone, 3g/L potassium dihydrogen phosphate, 2g/L dipotassium hydrogen phosphate and 3g/L magnesium sulfate;
according to the following steps of 1: 1.2, uniformly mixing the solid culture medium (except EM bacterial powder) and the nutrient solution according to the mass ratio, sealing the prepared culture medium, placing the culture medium in a high-pressure moist heat sterilization pot, and conventionally sterilizing for 40min at 121 ℃.
Inoculating the seed solution into the solid culture medium according to the inoculation amount of 10 wt% under the room temperature and aseptic condition, adding the EM bacterial powder with corresponding amount, uniformly mixing, then placing the inoculated culture medium into a cultivation room for cultivation, controlling the temperature at 17 ℃ and the humidity at 70%, and performing light-tight cultivation for 3-5 days until the spawn running is complete.
And then adjusting the illumination condition, controlling the illumination condition to be 300lux, and controlling the photoperiod light-dark ratio to be L16: d8, continuously controlling the culture temperature to be 17 ℃ and the humidity to be 70%, continuously culturing for 5-8 days to finish color conversion and form the stroma bud.
And then maintaining the illumination condition, continuously controlling the illumination condition to be 300lux, and controlling the photoperiod light-dark ratio to be L16: d8, controlling the culture temperature at 22 deg.C and humidity at 70%, continuously culturing for 15-20 days, collecting mature fruiting body, and oven drying.
Comparative example 1
The method for culturing cordyceps sobolifera sporocarp in the comparative example is the same as that in example 3, and only the EM bacterial powder is not contained in the solid matrix.
Comparative example 2
The method for culturing cordyceps sobolifera sporocarp in the comparative example is the same as that in example 3, and is only different from the method in that the solid matrix does not contain withered leaves of the hazy moon plant and broken leaves of the hazy moon plant.
Comparative example 3
The method for culturing cordyceps sobolifera sporocarp in the comparative example is the same as that in example 3, only the difference is that the solid matrix does not contain the EM bacterial powder, and the hazy moon plant dead leaves and the hazy moon plant leaf fragments.
Examples of the experiments
The fruiting bodies harvested in example 3 and comparative examples 1 to 3 were collected and pulverized into powder, and the crude polysaccharide was extracted by ultrasonic assisted-hot water extraction. Accurately weighing 1g of the above fruiting body powder, diluting with distilled water to constant volume of 100ml, placing into a test tube with a plug, performing ultrasonic extraction for 1h, and repeatedly reversing and mixing uniformly every 20 min; transferring into constant temperature water bath, extracting at 80 deg.C for 4 hr, cooling to room temperature, centrifuging the extractive solution at 14000rpm for 5min, and collecting supernatant as crude polysaccharide solution. The content of the crude polysaccharide was determined by a concentrated sulfuric acid-phenol method (see the preliminary study of artificial culture conditions of cordyceps sobolifera, zhangzewen, etc.), and the determined content of cordyceps sobolifera crude polysaccharide was recorded in table 1 below.
TABLE 1 Cordyceps cicadae Miquel polysaccharide content mg/g
| Numbering | Crude polysaccharide content mg/g fruiting body |
| Example 3 | 152.7 |
| Comparative example 1 | 140.2 |
| Comparative example 2 | 134.9 |
| Comparative example 3 | 118.3 |
Therefore, the method can effectively improve the content of the crude polysaccharide in the cordyceps sobolifera sporocarp by optimizing the solid culture medium.
It should be understood that the above examples are only for clarity of illustration and are not intended to limit the embodiments. Other variations and modifications will be apparent to persons skilled in the art in light of the above description. And are neither required nor exhaustive of all embodiments. And obvious variations or modifications therefrom are within the scope of the invention.
Claims (10)
1. An artificial cultivation method for improving the content of cordyceps sobolifera polysaccharide in cordyceps sobolifera sporocarp is characterized by comprising the steps of carrying out liquid strain culture in a liquid culture medium and carrying out sporocarp culture in a solid culture medium;
the solid culture medium comprises the following components in a mass ratio of 1: 0.8-1.5 of solid matrix and nutrient solution;
the solid matrix comprises the following components in parts by weight: 10-20 parts of rice, 20-30 parts of corn grit, 10-20 parts of bran, 10-20 parts of cicada pupa powder, 5-12 parts of withered leaves of an Ji hazy moon plant, 5-12 parts of broken leaves of the Ji hazy moon plant and 0.5-2 parts of EM (effective microorganism) powder;
the nutrient solution comprises the following components in percentage by mass: 20-30g/L of carbon source, 8-15g/L of nitrogen source, 3-8/L of inorganic salt and natural pH.
2. The artificial cultivation method for improving the content of cordyceps sobolifera polysaccharide in cordyceps sobolifera fruiting bodies according to claim 1, which is characterized in that:
the carbon source comprises glucose;
the nitrogen source comprises peptone;
the inorganic salt comprises a mixture of potassium dihydrogen phosphate, dipotassium hydrogen phosphate and magnesium sulfate.
3. The artificial cultivation method for increasing the content of cordyceps sobolifera polysaccharide in cordyceps sobolifera fruiting body according to claim 1 or 2, wherein the fruiting body cultivation step comprises a fungus growing step, a color changing step and a fruiting body management step.
4. The artificial cultivation method for increasing the content of cordyceps sobolifera polysaccharide in cordyceps sobolifera fruiting body according to any one of claims 1-3, wherein the control conditions of the fungus growing step comprise: culturing at 15-18 deg.C and humidity of 60-80% under dark condition.
5. The artificial cultivation method for increasing the content of cordyceps sobolifera polysaccharide in cordyceps sobolifera fruiting body according to any one of claims 1 to 4, wherein the control conditions of the color conversion step comprise: controlling the illumination condition to be 200-500lux, and controlling the photoperiod light-dark ratio to be L15-18: d6-9, the culture temperature is 15-18 ℃, and the humidity is 60-80%.
6. The artificial cultivation method for increasing the content of cordyceps sobolifera polysaccharide in cordyceps sobolifera fruiting body according to any one of claims 1 to 5, wherein the control conditions of the fruiting body management step comprise: controlling the illumination condition to be 200-500lux, and controlling the photoperiod light-dark ratio to be L15-18: d6-9, the culture temperature is 20-25 ℃, and the humidity is 60-80%.
7. The artificial cultivation method for improving the content of cordyceps sobolifera polysaccharide in cordyceps sobolifera fruiting bodies according to any one of claims 1 to 6, wherein the liquid culture medium comprises the following components in percentage by mass: 1-2% of glucose, 0.2-0.4% of yeast extract, 0.8-1.2% of water chestnut, 0.5-1% of egg white and MgSO 44 0.01-0.03％、KH2PO40.01-0.03%, natural pH.
8. The artificial cultivation method for increasing the content of cordyceps sobolifera polysaccharide in cordyceps sobolifera fruiting body according to any one of claims 1 to 7, wherein the conditions for liquid strain cultivation comprise: the fermentation temperature is controlled to be 20-25 ℃, and the stirring speed is 150 and 180 rpm.
9. The artificial cultivation method for increasing the content of cordyceps sobolifera polysaccharide in cordyceps sobolifera fruiting body according to any one of claims 1 to 8, further comprising the step of performing conventional activation on the strain before the liquid strain cultivation step.
10. Cordyceps cicadae fruiting body obtained by the cultivation method as claimed in any one of claims 1 to 9.
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| CN116138381A (en) * | 2022-12-16 | 2023-05-23 | 达州市命之源生物科技有限责任公司 | Cordyceps cicadae composition with liver and kidney nourishing effects and beverage prepared from Cordyceps cicadae composition |
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