CN113170699A - Graphene synergistic morchella culture medium and preparation method and application thereof - Google Patents
Graphene synergistic morchella culture medium and preparation method and application thereof Download PDFInfo
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- CN113170699A CN113170699A CN202010834104.8A CN202010834104A CN113170699A CN 113170699 A CN113170699 A CN 113170699A CN 202010834104 A CN202010834104 A CN 202010834104A CN 113170699 A CN113170699 A CN 113170699A
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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
- A01G18/20—Culture media, e.g. compost
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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
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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
- A01G18/40—Cultivation of spawn
Abstract
The invention provides a graphene synergistic morchella culture medium and a preparation method and application thereof. According to the method, the graphene powder which is rich in carboxyl and hydroxyl, good in water solubility, uniform in dispersion in water, strong in stability and not easy to aggregate is added into the culture medium for cultivating and producing the seeds of the morchella esculenta, so that the growth speed of the morchella esculenta mycelia is improved, sclerotia are formed in a large amount, and the final morchella esculenta sporocarp yield is improved; the cost of the culture medium is reduced, and the culture medium is beneficial to the acceptance of vast cultivators.
Description
Technical Field
The invention belongs to the field of artificial cultivation of edible fungi, and particularly relates to a graphene synergistic morchella culture medium and a preparation method and application thereof.
Background
Morchella (Morchella esculenta) belongs to Ascomycotina, class of Pediclocomycetes, order of Pedicellales, family of Morchellacaceae, genus of Morchella, is named as Morchella esculenta and Morchella esculenta due to its cellular head capable of being pregnant on the surface and its appearance extremely similar to Morchella esculenta. As a delicious dish loved by people, the morchella is rich in nutrition and complete in components, and the protein content of the morchella is higher than that of meat such as cattle, sheep, pigs and the like and is only inferior to that of soybeans; the health-care food also contains rich amino acids, fatty acids, polysaccharides, vitamins and a large amount of mineral elements, has unique flavor, has various medicinal effects of improving immunity, resisting viruses, resisting fatigue and the like, and has wide development and application prospects.
Graphene is currently the most promising application as a silicon substitute for fabricating ultra-micro transistors for the production of future supercomputers. By replacing silicon with graphene, the operating speed of a computer processor will be hundreds of times faster. In addition, graphene is almost completely transparent, absorbing only 2.3% of light. On the other hand, it is very dense and impenetrable by even the smallest gas atoms (helium atoms). These characteristics make it well suited as a raw material for transparent electronic products such as transparent touch displays, light-emitting panels, and solar panels. As a novel nano material which is the thinnest, the largest in strength and the strongest in electric conduction and heat conduction performance and is discovered at present, graphene is called as ' black gold ', which is the king of new materials ', scientists even predict that the graphene will ' completely change the 21 st century ', and the appearance of the graphene is very likely to bring up a subversive new technology and industrial revolution around the world. However, there are rarely reports of the application of graphene to non-material fields.
Disclosure of Invention
The present invention providesThe mother strain culture medium for the morchella comprises the following raw materials in parts by weight: 250 portions of 200-one potato, 20-25 portions of glucose, 20-25 portions of agar, 0.1-100 x 10 of graphene-3And (4) portions are obtained.
According to an embodiment of the invention, the potato is present in an amount of 210-240 parts, such as 200 parts, 210 parts, 220 parts, 230 parts, 240 parts or 250 parts.
According to an embodiment of the invention, the potatoes are potato peel free.
According to an embodiment of the invention, the glucose is present in an amount of 21-24 parts, such as 20 parts, 21 parts, 22 parts, 23 parts, 24 parts or 25 parts.
According to an embodiment of the invention, the agar is present in an amount of 21-24 parts, such as 20 parts, 21 parts, 22 parts, 23 parts, 24 parts or 25 parts.
According to the embodiment of the invention, the graphene is graphene powder. For example, the average sheet diameter of the graphene powder is 20 to 60nm, preferably 30 to 50nm, and exemplary is 30nm, 35nm, 40nm, 45nm, 50 nm. For example, the graphene has a monolayer thickness of 0.2-0.6nm, such as 0.3-0.5nm, exemplary 0.3nm, 0.32nm, 0.33nm, 0.334nm, 0.35nm, 0.4nm, 0.45nm, 0.5 nm. Preferably, the graphene is rich in carboxyl and hydroxyl groups, and is very soluble in water to form a uniform and stable solution. According to an embodiment of the present invention, the graphene is preferably prepared by an electrochemical method. For example, the electrochemical method comprises the following preparation processes: placing two electrode plates prepared by taking graphite powder as raw materials in an electrolytic oxidation tank, respectively connecting the electrode plates with a cathode and an anode, and carrying out electrolytic reaction by taking distilled water (preferably primary distilled water) as electrolyte to obtain graphene sol; and carrying out spray drying on the graphene sol to obtain graphene powder.
Preferably, the power supply is pulsed at a frequency of 30 to 50kHz, such as 34 to 45kHz, illustratively 40kHz, during electrolysis.
Preferably, the effective voltage during electrolysis is 8-15V, such as 10-14V, with 12V being exemplary.
Preferably, the effective current density during electrolysis is 40-60A/m2E.g. 45-55A/m2Exemplary is 50A/m2。
Preferably, the graphene sol has a mass concentration of 0.1-1%, such as 0.3-0.8%, exemplarily 0.5%.
Preferably, the temperature of the spray drying is 80-100 ℃, such as 85-95 ℃, exemplary 90 ℃.
According to an embodiment of the present invention, the content of the graphene is (0.5 to 80) × 10-3Parts, e.g. 0.1X 10-31 × 10 portions of-3Parts, 0.01 parts, 0.05 parts or 0.1 parts.
According to an embodiment of the present invention, the morchella mother culture medium further comprises water. Wherein the water content is 800-.
According to an embodiment of the present invention, the morchella mother strain culture medium comprises the following components: 200-250g of potato, 20-25g of glucose, 20-25g of agar, 0.1-100mg of graphene powder and 1000mL of water. Illustratively, the morchella stock culture medium comprises the following components: 200g of potato, 20g of glucose, 20g of agar, 0.1mg, 1mg, 10mg or 50mg of graphene and 1000mL of water.
The invention also provides a preparation method of the morchella mother strain culture medium, which comprises the following steps: sterilizing and cooling a mixture containing potato juice, agar, glucose, graphene and water to obtain the morchella mother strain culture medium. Preferably, the potato, agar, glucose, graphene and water have the proportions as described above.
According to an embodiment of the present invention, the process for preparing potato juice comprises: the peeled potatoes are boiled by adding water and then filtered to obtain juice. For example, the boiling is carried out under stirring, boiling for 20-40min, such as boiling for 25-35min, exemplary boiling for 30 min.
According to an embodiment of the present invention, the process for preparing the mixture comprising potato juice, agar, glucose, graphene and water comprises: adding agar into the potato juice, and heating until the agar is completely melted; and sequentially adding glucose and graphene into the mixture, and complementing the evaporated water to obtain the mixture.
According to an embodiment of the invention, the preparation method further comprises: the mixture is filled into a test tube while it is hot, sealed and sterilized. Further, the mixture was loaded in the test tube in an amount of 1/3 the volume of the test tube.
According to an embodiment of the invention, the sterilization is autoclaving, e.g. sterilization in an autoclave. Preferably, cool air is exhausted when the pressure finger rises to 0.04-0.06 pounds (e.g., 0.05 pounds), and after the finger returns to 0.00 pounds, the temperature is raised to 1.3-1.8 pounds (e.g., 1.5 pounds) for 20-40min (e.g., 20min, 30 min).
According to an embodiment of the present invention, the cooling is performed by naturally cooling to 40-50 ℃ (for example, 45 ℃), taking out the test tube, then placing the test tube on a slope, and continuing cooling to room temperature to solidify the culture medium.
According to an exemplary embodiment of the present invention, the preparation method of the morchella mother culture medium comprises the following steps:
(1) adding water into peeled potatoes for boiling, and then filtering to obtain juice to obtain potato juice;
(2) adding agar into the potato juice in the step (1), and heating until the agar is completely melted; sequentially adding glucose and graphene powder into the mixture, and complementing the evaporated moisture to obtain a mixture;
(3) and (3) putting the mixture obtained in the step (2) into a test tube, and carrying out autoclaving and cooling to obtain the morchella mother strain culture medium.
The invention also provides a morchella stock culture medium which comprises the following raw materials in parts by weight: 5-15 parts of rice hull, 0.5-3 parts of gypsum powder, 0-70 parts of boiled and swollen wheat grains, 5-15 parts of wood chips, 0-70 parts of caragana microphylla chips, 5-15 parts of humus soil and graphene (0.1-100) multiplied by 10-3Preparing; the consumption of the boiled and swollen wheat grains and the caragana microphylla crumbs is not 0 at the same time.
According to an embodiment of the invention, the graphene is added in the form of an aqueous graphene solution. For example, the concentration of the aqueous graphene solution is 50-100mg/L, such as 60-90mg/L, illustratively 50mg/L, 60mg/L, 70mg/L, 80mg/L, 90mg/L, or 100 mg/L. Preferably, the aqueous graphene solution contains almost no electrolyte.
According to an embodiment of the invention, the graphene has the meaning as described above.
According to an embodiment of the present invention, the content of the graphene is (0.5 to 80) × 10-3Parts, e.g. 0.1X 10-31 × 10 portions of-3Portion, 5X 10-3Parts, 0.01 parts, 0.05 parts or 0.1 parts.
According to an embodiment of the invention, the rice hull is present in an amount of 7.5 to 13 parts, illustratively 5 parts, 7.5 parts, 9 parts, 10 parts, 11 parts, 13 parts or 15 parts.
According to an embodiment of the invention, the content of the landplaster is 0.75-2 parts, exemplarily 0.5 part, 0.75 part, 1 part, 1.5 parts, 2 parts, 2.5 parts or 3 parts.
According to an embodiment of the invention, the swollen wheat kernels are swollen and do not exhibit whites. The water content is about 70-75%.
According to an embodiment of the invention, the content of the boiled-out wheat grains is 10-60 parts, exemplary 10 parts, 20 parts, 30 parts, 40 parts, 50 parts, 60 parts or 70 parts.
According to an embodiment of the invention, the wood chips are present in an amount of 7.5-13 parts, exemplarily 5 parts, 7.5 parts, 9 parts, 10 parts, 11 parts, 13 parts or 15 parts.
According to an embodiment of the present invention, the caragana microphylla crumb content is 10-60 parts, illustratively 10 parts, 20 parts, 30 parts, 40 parts, 50 parts, 60 parts or 70 parts.
According to an embodiment of the present invention, the caragana microphylla crumb has a length of 0.5-1cm and a width of 1-4 mm. Caragana microphylla is distributed in inner Mongolia, Ningxia and Gansu of China, grows in semi-fixed and fixed sand lands and is often the dominant species. Its branches and leaves can be used as green manure and feed, and is an excellent raw material for making green manure.
According to an embodiment of the invention, the content of the humus soil is 7.5-13 parts, exemplarily 5 parts, 7.5 parts, 9 parts, 10 parts, 11 parts, 13 parts or 15 parts.
According to an embodiment of the invention, the medium of the stock morchella species has a humidity of 60-70%, such as 63-68%, exemplarily 60%, 63%, 65%, 68% or 70%.
According to an embodiment of the invention, the pH of the stock culture of morchella is 7.0-8.0, such as 7.3-7.7, exemplary 7.0, 7.3, 7.5, 7.8 or 8.0.
The invention also provides a preparation method of the morchella stock culture medium, which comprises the following steps: mixing rice hulls, gypsum powder, boiled and expanded wheat grains, sawdust, caragana microphylla scraps, humus soil and a graphene water solution, adjusting the water content and the pH value of the mixture, and sterilizing to obtain the morchella stock culture medium.
Preferably, the rice hulls, the gypsum powder, the boiled and expanded wheat grains, the wood chips, the caragana microphylla scraps, the humus soil and the graphene aqueous solution are proportioned according to the proportion described above.
According to an embodiment of the present invention, the rice hulls, wood chips and caragana microphylla chips can be soaked in water until completely soaked, and then mixed with the gypsum powder, humus soil and graphene solution. For example, the soaking time is 24-48h, illustratively 24h, 30h, 36h, 40h, 48 h.
According to an embodiment of the invention, the water content and the pH have the ranges as described above.
According to an embodiment of the invention, the preparation method further comprises: bottling the mixture with water content and pH value, sealing, and sterilizing.
Wherein, the bottling is required to be consistent in tightness and not too tight, the bottle is preferably mounted on a flat shoulder, the surface is flattened, the culture medium stained outside the bottle wall is washed, and then a cotton plug is plugged for sealing.
According to an embodiment of the invention, the sterilization is autoclaving. For example, the sterilization pressure is 1-2 pounds, preferably 1.5 pounds. For example, the time for sterilization is 1 to 3 hours, preferably 2 hours.
According to an embodiment of the present invention, after the sterilization is completed, the medium is cooled to room temperature to obtain the morchella stock culture medium.
The invention also provides a morchella cultivar culture medium which comprises the following raw materials in parts by weight: 10-15 parts of rice husk and gypsum powder0.5-3 parts of boiled and swollen wheat grains 10-50 parts of sawdust 10-15 parts of caragana microphylla chips 10-60 parts of humus soil 5-15 parts of graphene (0.1-100) multiplied by 10-3And (4) portions are obtained.
According to an embodiment of the invention, the graphene is added in the form of an aqueous graphene solution. For example, the concentration of the aqueous graphene solution is 10-40mg/L, such as 20-30mg/L, illustratively 10mg/L, 20mg/L, 30mg/L, or 40 mg/L. Preferably, the aqueous graphene solution contains almost no electrolyte.
According to an embodiment of the present invention, the content of the graphene is (0.5 to 80) × 10-3Parts, e.g. 0.1X 10-31 × 10 portions of-3Portion, 5X 10-3Parts, 0.01 parts, 0.05 parts or 0.1 parts.
According to an embodiment of the invention, the graphene has the meaning as described above.
According to embodiments of the invention, the rice hulls are present in an amount of 11 to 14 parts, illustratively 10 parts, 11 parts, 12 parts, 13 parts, 14 parts, or 15 parts.
According to an embodiment of the invention, the content of the landplaster is 0.75-2 parts, exemplarily 0.5 part, 0.75 part, 1 part, 1.5 parts, 2 parts, 2.5 parts or 3 parts.
According to an embodiment of the invention, the swollen wheat kernels are swollen and do not exhibit whites. The water content is about 70-75%.
According to an embodiment of the invention, the content of the boiled-out wheat grains is 20-50 parts, exemplary 10 parts, 20 parts, 30 parts, 40 parts or 50 parts.
According to an embodiment of the invention, the wood chips are present in an amount of 11-14 parts, illustratively 10 parts, 11 parts, 12 parts, 13 parts, 14 parts or 15 parts.
According to an embodiment of the present invention, the caragana microphylla crumb content is 20-50 parts, illustratively 10 parts, 20 parts, 30 parts, 40 parts, 50 parts or 60 parts.
According to an embodiment of the present invention, the caragana microphylla crumb has a length of 0.5-1cm and a width of 1-4 mm. Caragana microphylla is distributed in inner Mongolia, Ningxia and Gansu of China, grows in semi-fixed and fixed sand lands and is often the dominant species. Its branches and leaves can be used as green manure and feed, and is an excellent raw material for making green manure.
According to an embodiment of the invention, the content of the humus soil is 7.5-13 parts, exemplarily 5 parts, 7.5 parts, 9 parts, 10 parts, 11 parts, 13 parts or 15 parts.
According to an embodiment of the invention, the humidity of the morchella cultivar medium is 60-70%, such as 63-68%, exemplarily 60%, 63%, 65%, 68% or 70%.
According to an embodiment of the invention, the pH of the morchella cultivar medium is 7.0-8.0, such as 7.3-7.7, exemplary 7.0, 7.3, 7.5, 7.8 or 8.0.
The invention also provides a preparation method of the morchella cultivar culture medium, which comprises the following steps: mixing rice hulls, gypsum powder, boiled and expanded wheat grains, sawdust, caragana microphylla scraps, humus soil and a graphene water solution, adjusting the water content and the pH value of the mixture, and sterilizing to obtain the morchella stock culture medium.
Preferably, the rice hulls, the gypsum powder, the boiled and expanded wheat grains, the wood chips, the caragana microphylla scraps, the humus soil and the graphene aqueous solution are proportioned according to the proportion described above.
According to an embodiment of the present invention, the rice hulls, wood chips and caragana microphylla chips can be soaked in water until completely soaked, and then mixed with the gypsum powder, humus soil and graphene solution. For example, the soaking time is 24-48h, illustratively 24h, 30h, 36h, 40h, 48 h.
According to an embodiment of the invention, the water content and the pH have the ranges as described above.
According to an embodiment of the invention, the preparation method further comprises: bagging the mixture with the adjusted water content and pH value, and sterilizing.
According to an embodiment of the invention, the sterilization is autoclaving, e.g. sterilization in an autoclave. Preferably, cool air is exhausted when the pressure finger rises to 0.04-0.06 pounds (e.g., 0.05 pounds), and after the finger returns to 0.00 pounds, the temperature is increased to 1.3-1.8 pounds (e.g., 1.5 pounds) and maintained for 20-40 minutes (e.g., 30 minutes).
According to an embodiment of the invention, after the sterilization is completed, the medium is cooled to room temperature to obtain the morchella cultivar medium.
The invention also provides a morchella nutrition bag which comprises the following raw materials in parts by weight: 5-15 parts of bran, 0.5-3 parts of gypsum powder, 20-40 parts of sawdust, 5-15 parts of millet shell, 40-60 parts of caragana microphylla scraps and graphene (0.1-100) multiplied by 10-3And (4) portions are obtained.
According to an embodiment of the invention, the bran is present in an amount of 7.5-13 parts, illustratively 5 parts, 7.5 parts, 9 parts, 10 parts, 11 parts, 13 parts or 15 parts.
According to an embodiment of the invention, the content of the landplaster is 0.75-2 parts, exemplarily 0.5 part, 0.75 part, 1 part, 1.5 parts, 2 parts, 2.5 parts or 3 parts.
According to an embodiment of the invention the wood chips are present in an amount of 25-35 parts, such as 20 parts, 25 parts, 30 parts, 35 parts or 40 parts.
According to an embodiment of the invention, the content of the millet husks is 7.5-13 parts, exemplarily 5 parts, 7.5 parts, 9 parts, 10 parts, 11 parts, 13 parts or 15 parts.
According to an embodiment of the present invention, the caragana microphylla crumb content is 45-55 parts, such as 40 parts, 45 parts, 50 parts, 55 parts or 60 parts.
According to an embodiment of the invention, the graphene is added in the form of an aqueous graphene solution. For example, the concentration of the aqueous graphene solution is 10-40mg/L, such as 20-30mg/L, illustratively 10mg/L, 20mg/L, 30mg/L, or 40 mg/L.
According to an embodiment of the present invention, the content of the graphene is (0.5 to 80) × 10-3Parts, e.g. 0.1X 10-31 × 10 portions of-3Portion, 5X 10-3Parts, 0.01 parts, 0.05 parts or 0.1 parts.
According to an embodiment of the invention, the graphene has the meaning as described above.
According to an embodiment of the invention, the humidity of the morchella nutrition bag is 60-70%, such as 63-68%, exemplarily 60%, 63%, 65%, 68% or 70%.
According to an embodiment of the invention, the pH of the morchella nutrition bag is 7.0-8.0, such as 7.3-7.7, exemplary 7.0, 7.3, 7.5, 7.8 or 8.0.
The invention also provides a preparation method of the morchella nutrition bag, which comprises the following steps: mixing bran, gypsum powder, sawdust, caragana microphylla scraps, millet shell and graphene aqueous solution, adjusting the water content and pH value of the mixture, and sterilizing to obtain the morchella stock culture medium.
Preferably, the bran, landplaster, wood flour, caragana microphylla chips, millet shell and graphene are in the proportions as described above.
According to an embodiment of the present invention, the bran, the wood flour, the millet shell and the caragana microphylla scraps may be soaked in water until completely soaked, and then mixed with the gypsum powder and the graphene aqueous solution. For example, the soaking time is 24-48h, illustratively 24h, 30h, 36h, 40h, 48 h.
According to an embodiment of the invention, the water content and the pH have the ranges as described above.
According to an embodiment of the invention, the preparation method further comprises: and bagging the mixture with the adjusted water content and pH value, sealing, and sterilizing.
According to an embodiment of the invention, the sterilization is autoclaving. For example, the sterilization pressure is 1-2 pounds, preferably 1.5 pounds. For example, the time for sterilization is 1 to 3 hours, preferably 2 hours.
According to an embodiment of the invention, after the sterilization is completed, the morchella nutrition bag is cooled to room temperature to obtain the morchella nutrition bag.
In the present invention, the water in the graphene aqueous solution may be purified water, distilled water, or tap water. Preferably, purified water or distilled water is selected as water in the graphene aqueous solution when the mother culture medium and/or the stock culture medium is prepared, and tap water is selected as water in the graphene aqueous solution when the culture medium is prepared.
The invention also provides application of any one of the culture media or the combination thereof in cultivation and seed production of morchella esculenta.
The invention also provides a method for culturing the morchella mother strain, which comprises the step of culturing the morchella mother strain by adopting the morchella mother strain culture medium. Preferably, the morchella mother strain is obtained by tissue separation, strictly purified, cultured at 16-20 ℃ in a dark place, and refrigerated for storage after the mother strain grows.
The invention also provides a method for culturing the stock of the morchella esculenta, which comprises the step of culturing the stock of the morchella esculenta by adopting the stock culture medium of the morchella esculenta.
The invention also provides a morel inoculation and culture method, which comprises the step of inoculating and culturing by adopting the morel cultivar culture medium. Preferably, the inoculation is carried out in a sterilized inoculation hood. Preferably, the temperature of the culture is 16-20 ℃ and the culture time is 18-22 days (preferably 20 days). Preferably, intense light stimulation and excessive temperature are avoided as much as possible during the culturing.
The invention also provides a using method of the morel nutrition bag, which comprises the step of using the morel nutrition bag in the morel cultivation process. Preferably, the substances in the morchella nutrient bag are fully contacted with the soil of a morchella cultivation field.
According to an embodiment of the invention, the method of use comprises the steps of: placing the toadstool nutrition bag in a toadstool cultivation field, arranging an opening on the nutrition bag, horizontally placing one side with the opening on the ground, and pressurizing to enable the side with the opening to fully contact with soil.
According to the embodiment of the invention, the toadstool nutrition bag is placed in a shape like a Chinese character 'pin' in a toadstool cultivation field. Preferably the bag to bag spacing is 30 to 50cm, for example 35 to 45cm, illustratively 40 cm.
The invention also provides a method for cultivating and producing seeds of morchella esculenta, which comprises the process of cultivating and producing seeds of morchella esculenta by using any one of the culture mediums or the combination thereof, or any one of the cultivation, inoculation and/or use methods.
The invention has the beneficial effects that:
the inventors surprisingly found that adding graphene powder which is rich in carboxyl and hydroxyl, good in water solubility, uniform in dispersion in water, strong in stability and not easy to aggregate into a culture medium for cultivating and producing the seeds of morchella esculenta improves the growth speed of morchella esculenta hyphae, enables sclerotia to be formed in a large amount and has a promotion effect on the final morchella esculenta fruiting body yield; the cost of the culture medium is reduced, and the culture medium is beneficial to the acceptance of vast cultivators.
Drawings
FIG. 1 shows growth of Morchella esculenta in synergistic mother culture media with different graphene contents.
FIG. 2 shows the formation of sclerotium of Morchella esculenta cultured in graphene enhanced stock culture medium (left) and stock culture medium without graphene (right).
Detailed Description
The technical solution of the present invention will be further described in detail with reference to specific embodiments. It is to be understood that the following examples are only illustrative and explanatory of the present invention and should not be construed as limiting the scope of the present invention. All the technologies realized based on the above-mentioned contents of the present invention are covered in the protection scope of the present invention.
Unless otherwise indicated, the raw materials and reagents used in the following examples are all commercially available products or can be prepared by known methods.
In the following examples, the preparation of graphene sols was carried out: graphite powder with different meshes is used as a raw material, the graphite powder is pressed into plate-shaped electrodes with the size of 300 multiplied by 200 multiplied by 20mm through an isostatic pressing process, two plate electrodes are placed into an electrolytic oxidation tank and are respectively connected with a cathode and an anode, primary distilled water is poured into the electrolytic oxidation tank to be used as electrolyte, the pulse frequency of a power supply is adjusted to be 40kHz, the effective voltage is 12V, and the effective current density is controlled to be 50A/m2And reacting for 150 hours to obtain the graphene sol with the mass concentration of 0.5%.
The preparation process of the graphene powder comprises the following steps: and (3) spray-drying the obtained sol at 90 ℃ to obtain black graphene powder.
Parameters of the graphene powder: the average sheet diameter is 40nm, and the single-layer thickness is 0.334 nm; is rich in carboxyl and hydroxyl, is very soluble in water, and forms a uniform and stable solution in the water.
Example 1
The mother strain culture medium of morchella comprises: 200g of peeled potato, 20g of glucose powder, 20g of agar, 1000mL of water and 0.1mg of graphene powder.
The preparation process comprises the following steps: adding 1000mL of clear water into peeled potatoes, boiling while stirring for 30min, filtering with four layers of gauze to obtain juice, adding agar, continuously heating until the materials are completely melted, adding glucose, adding 0.1mg of graphene powder, supplementing the evaporated water to 1000mL, hot-filling into test tubes with the specification of 18 x 180mm, filling 1/3 per test tube, plugging with a cotton plug, bundling every 10 test tubes, and placing into an autoclave for sterilization;
the sterilization method of the mother strain culture medium of the morchella comprises the following steps: heating the autoclave, discharging cold air when the pressure pointer rises to 0.05 pound, continuously heating to 1.5 pound after the pointer returns to 0.00 pound, maintaining for 30min, naturally cooling the autoclave, taking out the test tube when the test tube is hot, and placing the test tube into an inclined plane, wherein the length of the culture medium is preferably about 2/3 of the length of the test tube.
Example 2
The mother strain culture medium of morchella comprises: 200g of peeled potato, 20g of glucose powder, 20g of agar, 1000mL of water and 1mg of graphene powder.
The preparation process comprises the following steps: adding 1000mL of clear water into peeled potatoes, boiling while stirring for 30min, filtering with four layers of gauze to obtain juice, adding agar, continuously heating until the materials are completely melted, adding glucose, adding 1mg of graphene powder, supplementing the evaporated water to 1000mL, putting the obtained product into test tubes with the specification of 18 x 180mm while the obtained product is hot, filling 1/3 per test tube, plugging cotton plugs, putting 10 bundles of the test tubes into an autoclave for sterilization;
the sterilization method of the mother strain culture medium of the morchella comprises the following steps: heating the autoclave, discharging cold air when the pressure pointer rises to 0.05 pound, continuously heating to 1.5 pound after the pointer returns to 0.00 pound, maintaining for 30min, naturally cooling the autoclave, taking out the test tube when the test tube is hot, and placing the test tube into an inclined plane, wherein the length of the culture medium is preferably about 2/3 of the length of the test tube.
Example 3
The mother strain culture medium of morchella comprises: 200g of peeled potato, 20g of glucose powder, 20g of agar, 1000mL of water and 10mg of graphene powder.
The preparation process comprises the following steps: adding 1000mL of clear water into peeled sheep potatoes, boiling while stirring for 30min, filtering with four layers of gauze to obtain juice, adding agar, continuously heating until the materials are completely melted, adding glucose, adding 10mg of graphene powder, supplementing the evaporated water to 1000mL, putting the obtained product into test tubes with the specification of 18 x 180mm while the obtained product is hot, filling 1/3 per test tube, plugging cotton plugs, bundling 10 test tubes, and putting the test tubes into an autoclave for sterilization;
the sterilization method of the mother strain culture medium of the morchella comprises the following steps: heating the autoclave, discharging cold air when the pressure pointer rises to 0.05 pound, continuously heating to 1.5 pound after the pointer returns to 0.00 pound, maintaining for 30min, naturally cooling the autoclave, taking out the test tube when the test tube is hot, and placing the test tube into an inclined plane, wherein the length of the culture medium is preferably about 2/3 of the length of the test tube.
Example 4
The mother strain culture medium of morchella comprises: 200g of peeled potato, 20g of glucose powder, 20g of agar, 1000mL of water and 50mg of graphene powder.
The preparation process comprises the following steps: adding 1000mL of clear water into peeled potatoes, boiling while stirring for 30min, filtering with four layers of gauze to obtain juice, adding agar, continuously heating until the materials are completely melted, adding glucose, adding 50mg of graphene powder, supplementing the evaporated water to 1000mL, putting the obtained product into test tubes with the specification of 18 x 180mm while the obtained product is hot, filling 1/3 per test tube, plugging cotton plugs, putting 10 bundles of the test tubes into an autoclave for sterilization;
the sterilization method of the mother strain culture medium of the morchella comprises the following steps: heating the autoclave, discharging cold air when the pressure pointer rises to 0.05 pound, continuously heating to 1.5 pound after the pointer returns to 0.00 pound, maintaining for 30min, naturally cooling the autoclave, taking out the test tube when the test tube is hot, and placing the test tube into an inclined plane, wherein the length of the culture medium is preferably about 2/3 of the length of the test tube.
Example 5
Different from the embodiment, the content of the graphene powder in the mother culture medium of the morchella esculenta is 100 mg.
Example 6
The toadstool stock culture medium comprises: 10% of rice hull, 1% of gypsum powder, 70% of cooked and swollen wheat grains, 10% of wood chips and 9% of humus soil, wherein a graphene solution is added in 50mg/L to prepare a culture medium with the humidity of 65% and the pH of the culture medium of 7-8.
The graphene solution is obtained by diluting graphene sol by 100 times with distilled water or purified water.
The preparation process comprises the following steps: firstly soaking rice hulls and wood chips in water for 24-48h, then diluting the graphene sol by 100 times to obtain a 50mg/L graphene solution, finally uniformly mixing the soaked rice hulls and wood chips with boiled and expanded wheat grains, gypsum powder and humus soil, adding the graphene solution, and bottling when the water content of the culture medium reaches 65%. Bottling, pressing the surface flat, washing off the culture medium contaminated outside the bottle wall, and sealing with a cotton plug; the sterilization method of the stock culture medium of the morchella comprises the following steps: autoclaving was maintained at 1.5 lbs pressure for 2h and cooling to room temperature.
Example 7
The toadstool stock culture medium comprises: 10% of rice hull, 1% of gypsum powder, 70% of boiled and swollen wheat grains, 10% of wood chips and 9% of humus soil, and 100mg/L of graphene solution is added to prepare a culture medium with the humidity of 65%. The pH of the culture medium is 7-8.
The graphene solution is obtained by diluting the graphene sol by 50 times with distilled water or purified water.
The preparation process comprises the following steps: firstly soaking rice hulls and wood chips in water for 24-48h, then diluting the graphene sol by 50 times to obtain a 100mg/L graphene solution, finally uniformly mixing the soaked rice hulls and wood chips with boiled and expanded wheat grains, gypsum powder and humus soil, adding the graphene solution, and bottling when the water content of the culture medium reaches 65%. Bottling, pressing the surface flat, washing off the culture medium contaminated outside the bottle wall, and sealing with a cotton plug; the sterilization method of the stock culture medium of the morchella comprises the following steps: autoclaving was maintained at 1.5 lbs pressure for 2h, cooled to room temperature, and removed for use.
Example 8
The toadstool stock culture medium comprises: 10% of rice hull, 1% of gypsum powder, 10% of wood chips, 70% of caragana microphylla scraps, 9% of humus soil, and 50mg/L of graphene solution is added to prepare the humidity of the culture medium to be 65%. The pH of the culture medium is 7-8.
The graphene solution is obtained by diluting graphene sol by 100 times with distilled water or purified water.
The preparation process comprises the following steps: soaking the rice hulls, the caragana microphylla fragments and the saw dust in water for 24-48h, then diluting the graphene sol by 100 times to obtain a 50mg/L graphene solution, finally uniformly mixing the soaked rice hulls, saw dust, caragana microphylla fragments, gypsum powder and humus soil, adding the graphene solution, and bottling when the water content of the culture medium reaches 65%. Bottling, pressing the surface flat, washing off the culture medium contaminated outside the bottle wall, and sealing with a cotton plug; the sterilization method of the stock culture medium of the morchella comprises the following steps: autoclaving was maintained at 1.5 lbs pressure for 2h, cooled to room temperature, and removed for use.
Example 9
The toadstool stock culture medium comprises: 10% of rice hull, 1% of gypsum powder, 10% of wood chips, 70% of caragana microphylla scraps, 9% of humus soil, and 100mg/L of graphene solution is added to prepare the humidity of the culture medium to be 65%. The pH of the culture medium is 7-8.
The graphene solution is obtained by diluting the graphene sol by 50 times with distilled water or purified water.
The preparation process comprises the following steps: soaking the rice hulls, the caragana microphylla fragments and the saw dust in water for 24-48h, then diluting the graphene sol by 50 times to obtain a 100mg/L graphene solution, finally uniformly mixing the soaked rice hulls, saw dust, caragana microphylla fragments, gypsum powder and humus soil, adding the graphene solution, and bottling when the water content of the culture medium reaches 65%. Bottling, pressing the surface flat, washing off the culture medium contaminated outside the bottle wall, and sealing with a cotton plug; the sterilization method of the stock culture medium of the morchella comprises the following steps: autoclaving was maintained at 1.5 lbs pressure for 2h, cooled to room temperature, and removed for use.
Example 10
The culture medium of the morchella cultivar comprises: 15% of rice hull, 1% of gypsum powder, 50% of boiled and expanded wheat grains, 15% of wood chips, 10% of caragana microphylla scraps and 9% of humus soil, and adding 10mg/L of graphene solution to adjust the humidity of the culture medium to 65%. The pH of the culture medium is 7-8.
The graphene solution is obtained by diluting graphene sol by 500 times with tap water.
The preparation process comprises the following steps: soaking rice hulls, caragana microphylla scraps and wood chips in water for 24-48h, diluting graphene sol by 500 times to obtain 10mg/L graphene solution, uniformly mixing the soaked rice hulls, caragana microphylla scraps, wood chips, boiled and expanded wheat grains, gypsum powder and humus soil, adding the graphene solution, bagging the culture medium in polypropylene bags when the water content of the culture medium reaches 65%, wherein the bag specification is 14 multiplied by 28cm, and each bag is filled with 0.5-0.6 kg of materials. Autoclaving and sterilizing Morchella mother culture medium, and inoculating when temperature is reduced to below 28 deg.C. Inoculating in a seed box, and sterilizing with 5g/m of "Youchlorin disinfectant powder3Igniting and fumigating for 30min, then inoculating, placing in an environment of 16-18 ℃ after inoculating, culturing for about 20 days, and directly using for cultivation after the mycelium is full of bags.
Example 11
The culture medium of the morchella cultivar comprises: 15% of rice hull, 1% of gypsum powder, 50% of boiled and expanded wheat grains, 15% of wood chips, 10% of caragana microphylla scraps and 9% of humus soil, and 40mg/L of graphene solution is added to prepare the humidity of the culture medium to 65%. The pH of the culture medium is 7-8.
The graphene solution is obtained by diluting graphene sol by 125 times with tap water.
The preparation process comprises the following steps: soaking rice hulls, caragana microphylla scraps and wood chips in water for 24-48h, diluting a graphene stock solution by 125 times to obtain a 40mg/L graphene solution, uniformly mixing the soaked rice hulls, caragana microphylla scraps, wood chips, boiled and expanded wheat grains, gypsum powder and humus soil, adding the graphene solution, bagging the culture medium in a polypropylene bag when the water content of the culture medium reaches 65%, wherein the bag is 14 x 28cm in specification, and each bag is filled with 0.5-0.6 kg of materials. Autoclaving and sterilizing Morchella mother culture medium, and inoculating when temperature is reduced to below 28 deg.C. Inoculating in a seed box, and sterilizing with 5g/m of "Youchlorin disinfectant powder3Igniting and fumigating for 30min, then inoculating, placing in an environment of 16-18 ℃ after inoculating, culturing for about 20 days, and directly using for cultivation after the mycelium is full of bags.
Example 12
The culture medium of the morchella cultivar comprises: 10% of rice hull, 1% of gypsum powder, 10% of boiled and swollen wheat grains, 10% of wood chips, 60% of caragana microphylla scraps and 9% of humus soil, and adding 10mg/L of graphene solution to adjust the humidity of the culture medium to 65%. The pH value of the culture medium is 7-8.
The graphene solution is obtained by diluting graphene sol by 500 times with tap water.
The preparation process comprises the following steps: soaking rice hulls, caragana microphylla scraps and wood chips in water for 24-48h, diluting a graphene stock solution by 500 times to obtain a 10mg/L graphene solution, uniformly mixing the soaked rice hulls, caragana microphylla scraps, wood chips, boiled and expanded wheat grains, gypsum powder and humus soil, adding the graphene solution, bagging the culture medium in a polypropylene bag when the water content of the culture medium reaches 65%, wherein the bag is 14 x 28cm in specification, and each bag is filled with 0.5-0.6 kg of materials. Autoclaving and sterilizing Morchella mother culture medium, and inoculating when temperature is reduced to below 28 deg.C. Inoculating in a seed box, and sterilizing with 5g/m of "Youchlorin disinfectant powder3Igniting and fumigating for 30min, then inoculating, placing in an environment of 16-18 ℃ after inoculating, culturing for about 20 days, and directly using for cultivation after the mycelium is full of bags.
Example 13
The culture medium of the morchella cultivar comprises: 10% of rice hull, 1% of gypsum powder, 10% of boiled and swollen wheat grains, 10% of wood chips, 60% of caragana microphylla scraps and 9% of humus soil, and 40mg/L of graphene solution is added to prepare the humidity of the culture medium to 65%. The pH of the culture medium is 7-8.
The graphene solution is obtained by diluting graphene sol by 125 times with tap water.
The preparation process comprises the following steps: soaking rice hulls, caragana microphylla scraps and wood chips in water for 24-48h, diluting a graphene stock solution by 125 times to obtain a 40mg/L graphene solution, uniformly mixing the soaked rice hulls, caragana microphylla scraps, wood chips, boiled and expanded wheat grains, gypsum powder and humus soil, adding the graphene solution, bagging the culture medium in a polypropylene bag when the water content of the culture medium reaches 65%, wherein the bag is 14 x 28cm in specification, and each bag is filled with 0.5-0.6 kg of materials. Autoclaving and processing lamb tripeThe sterilization method of the mother culture medium comprises the steps of sterilizing, and then inoculating when the temperature is reduced to be below 28 ℃. Inoculating in a seed box, and sterilizing with 5g/m of "Youchlorin disinfectant powder3Igniting and fumigating for 30min, then inoculating, placing in an environment of 16-18 ℃ after inoculating, culturing for about 20 days, and directly using for cultivation after the mycelium is full of bags.
Example 14
The toadstool nutrition bag comprises: 10% of bran, 1% of gypsum powder, 30% of sawdust, 9% of millet shell and 50% of caragana microphylla scraps, and adding 10mg/L of graphene solution to adjust the humidity of the culture medium to 65%. The pH is 7-8.
The preparation process comprises the following steps: firstly, soaking bran, sawdust, millet shell and caragana microphylla scraps in water for 24-48h, then diluting graphene sol by 500 times with tap water to obtain a 10mg/L graphene solution, finally uniformly mixing the soaked bran, sawdust, millet shell, caragana microphylla scraps and gypsum powder, adding the treated graphene solution, and bagging when the water content of a culture medium reaches 65%, wherein the specification of the nutrition bag is selected by filling polypropylene or polyethylene plastic bags of 13 x 26cm, filling the mixed culture medium into bags, wherein the bags are filled with about 0.5-0.6 kg of culture medium, the tightness is required to be proper, and after filling, fastening and sterilizing the bag openings;
the sterilization method of the morchella nutrition bag comprises the following steps: if autoclaving is used, the mixture is held at 1.5 lbs for 2 hours, cooled and removed for further use.
Example 15
The toadstool nutrition bag comprises: 10% of bran, 1% of gypsum powder, 30% of sawdust, 9% of millet shell and 50% of caragana microphylla scraps, and 40mg/L of graphene solution is added to adjust the humidity of the culture medium to 65%. The pH is natural.
The preparation process comprises the following steps: firstly, soaking bran, sawdust, millet shell and caragana microphylla scraps in water for 24-48h, then diluting graphene sol by 125 times with tap water to obtain 40mg/L graphene solution, finally uniformly mixing the soaked bran, sawdust, millet shell, caragana microphylla scraps and gypsum powder, adding the treated graphene solution, and bagging when the water content of a culture medium reaches 65%, wherein the specification of the nutrition bag is selected by filling polypropylene or polyethylene plastic bags of 13 x 26cm, filling the stirred culture medium into the bags, wherein the bags are filled with about 0.5-0.6 kg of culture medium, the tightness is required to be proper, and after filling, fastening and sterilizing the bag openings;
the sterilization method of the morchella nutrition bag comprises the following steps: if autoclaving is used, the mixture is held at 1.5 lbs for 2 hours, cooled and removed for further use.
Test example 1
And (3) testing the growth condition of the morchella in mother culture media with different graphene contents:
the experimental procedure was as follows: 1) mother culture media with different graphene contents were prepared according to examples 1 to 5, and a mother culture medium without graphene was prepared at the same time as a control; 2) taking out the preserved Morchella strains, and inoculating the Morchella strains with the same amount to the central part of each culture medium by using a puncher under the aseptic condition; 3) after the step 2) is finished, placing the culture dish in an artificial climate box, setting the temperature to be 18 ℃, setting the relative humidity to be 80%, culturing in a dark place, and periodically testing various indexes. The growth of morchella mycelia in the synergistic mother culture media with different graphene contents is shown in table 1 and fig. 1. Wherein, the group A is a control group without adding graphene powder, B-F respectively correspond to the mother culture medium provided in the embodiment 1-5, and the content of graphene in B-F is respectively 0.1mg/L, 1mg/L, 10mg/L, 50mg/L and 100 mg/L. It can be seen that in the mother culture medium added with the graphene powder, the growth speed of morchella mycelium is obviously improved, and a large amount of sclerotia is formed.
TABLE 1 growth of morchella mycelium in synergistic mother culture media with different graphene contents
Test example 2 growth of Morchella in stock culture
The experimental procedure was as follows: 1) graphene stock culture medium was prepared according to example 7, while stock culture medium without graphene was prepared as a control group; 2) selecting soybean grains with the size of a preserved morchella strain by using an inoculating hook under an aseptic condition, and inoculating the soybean grains into a culture medium, wherein two samples are arranged in parallel in each group of culture medium; 3) and after inoculation, the toadstool is cultured for 20 days in a dark environment at the temperature of 16-18 ℃ and then photographed, and the influence of the stock culture medium containing graphene and the stock culture medium without graphene on the growth and the sclerotia of the toadstool is compared.
The growth situation is shown in fig. 2 (two bottles on the left are graphene synergistic stock culture medium, two bottles on the right are control group): the growth condition of morchella in the graphene synergistic stock culture medium is obviously superior to that of a control group.
According to the invention, by adding the formula of the four culture media for cultivating and producing the seeds of the morchella esculenta, the growth speed of morchella esculenta mycelia is improved, sclerotia are formed in a large amount, and the final morchella esculenta sporocarp yield is improved. The cost of the culture medium is reduced, and the culture medium is beneficial to the acceptance of vast cultivators.
The embodiments of the present invention have been described above. However, the present invention is not limited to the above embodiment. Any modification, equivalent replacement, or improvement made within the spirit and principle of the present invention should be included in the protection scope of the present invention.
Claims (10)
1. The mother strain culture medium for the morchella comprises the following raw materials in parts by weight: 250 portions of 200-one potato, 20-25 portions of glucose, 20-25 portions of agar, 0.1-100 x 10 of graphene-3And (4) portions are obtained.
Preferably, the graphene is graphene powder. For example, the average sheet diameter of the graphene powder is 20 to 60 nm. For example, the graphene has a monolayer thickness of 0.2 to 0.6 nm. Preferably, the graphene is rich in carboxyl and hydroxyl groups.
Preferably, the graphene is preferably prepared by an electrochemical method. Preferably, the electrochemical method comprises the following preparation processes: placing two electrode plates prepared by taking graphite powder as raw materials in an electrolytic oxidation tank, respectively connecting the electrode plates with a cathode and an anode, taking distilled water as electrolyte, and carrying out electrolytic reaction to obtain graphene sol; and carrying out spray drying on the graphene sol to obtain graphene powder.
Preferably, the morchella mother strain culture medium further comprises water.
Preferably, the morchella mother strain culture medium comprises the following components: 200-250g of potato, 20-25g of glucose, 20-25g of agar, 0.1-100mg of graphene powder and 1000mL of water.
Preferably, the preparation method of the morchella mother culture medium comprises the following steps: sterilizing and cooling a mixture containing potato juice, agar, glucose, graphene and water to obtain the morchella mother strain culture medium. Preferably, the preparation method further comprises: the mixture was filled into test tubes while hot, sealed and sent for sterilization. Further, the mixture was loaded in the test tube in an amount of 1/3 the volume of the test tube.
2. A toadstool stock culture medium comprises the following raw materials in parts by weight: 5-15 parts of rice hull, 0.5-3 parts of gypsum powder, 0-70 parts of boiled and swollen wheat grains, 5-15 parts of wood chips, 0-70 parts of caragana microphylla chips, 5-15 parts of humus soil and graphene (0.1-100) multiplied by 10-3Preparing; the consumption of the boiled and swollen wheat grains and the caragana microphylla crumbs is not 0 at the same time.
Preferably, the graphene is added in the form of an aqueous graphene solution. For example, the concentration of the graphene aqueous solution is 50-100 mg/L.
Preferably, the graphene has the meaning as defined in claim 1.
Preferably, the humidity of the toadstool stock culture medium is 60-70%.
Preferably, the pH of the toadstool stock culture medium is 7.0-8.0.
Preferably, the preparation method of the morchella stock culture medium comprises the following steps: mixing rice hulls, gypsum powder, boiled and expanded wheat grains, sawdust, caragana microphylla scraps, humus soil and a graphene water solution, adjusting the water content and the pH value of the mixture, and sterilizing to obtain the morchella stock culture medium. Preferably, the preparation method further comprises: bottling the mixture with adjusted water content and pH, sealing, and sterilizing; and after the sterilization is finished, cooling to room temperature to obtain the morchella stock culture medium.
3. A toadstool cultivar culture medium comprises the following raw materials in parts by weight: 10-15 parts of rice husk and gypsum0.5-3 parts of flour, 10-50 parts of boiled and swollen wheat grains, 10-15 parts of sawdust, 10-60 parts of caragana microphylla scraps, 5-15 parts of humus soil and graphene (0.1-100) multiplied by 10-3And (4) portions are obtained.
Preferably, the graphene is added in the form of an aqueous graphene solution. For example, the concentration of the graphene aqueous solution is 10-40 mg/L.
Preferably, the graphene has the meaning as defined in claim 1.
Preferably, the humidity of the morchella cultivar culture medium is 60-70%.
Preferably, the pH of the morchella cultivar culture medium is 7.0-8.0.
Preferably, the preparation method of the morchella cultivar culture medium comprises the following steps: mixing rice hulls, gypsum powder, boiled and expanded wheat grains, sawdust, caragana microphylla scraps, humus soil and a graphene water solution, adjusting the water content and the pH value of the mixture, and sterilizing to obtain the morchella stock culture medium. Preferably, the preparation method further comprises: bagging the mixture with the adjusted water content and pH value, and sterilizing; and after the sterilization is finished, cooling to room temperature to obtain the morchella cultivar culture medium.
4. The morchella nutrition bag comprises the following raw materials in parts by weight: 5-15 parts of bran, 0.5-3 parts of gypsum powder, 20-40 parts of sawdust, 5-15 parts of millet shell, 40-60 parts of caragana microphylla scraps and graphene (0.1-100) multiplied by 10-3And (4) portions are obtained.
Preferably, the graphene is added in the form of an aqueous graphene solution. For example, the concentration of the graphene aqueous solution is 10-40 mg/L. Preferably, the graphene has the meaning as defined in claim 1.
Preferably, the humidity of the toadstool nutrition bag is 60-70%.
Preferably, the pH of the morchella nutrition bag is 7.0-8.0.
Preferably, the preparation method of the morchella nutrition bag comprises the following steps: mixing bran, gypsum powder, sawdust, caragana microphylla scraps, millet shell and graphene aqueous solution, adjusting the water content and pH value of the mixture, and sterilizing to obtain the morchella stock culture medium.
Preferably, the preparation method further comprises: bagging the mixture with the adjusted water content and pH value, sealing, and sterilizing; and after the sterilization is finished, cooling to room temperature to obtain the morchella esculenta nutrition bag.
5. Use of any one of the culture media of claims 1-4 or a combination thereof for the production of seeds by morchella cultivation.
6. A method for culturing Morchella esculenta mother strain, comprising culturing Morchella esculenta mother strain with the Morchella esculenta mother strain culture medium of claim 1.
7. A method for culturing a Morchella stock culture, comprising culturing the Morchella stock culture with the Morchella stock culture medium of claim 2.
8. A Morchella inoculation and culture method, comprising inoculating and culturing with the Morchella cultivar culture medium of claim 3.
9. A use method of the Morchella esculenta nutrition bag, comprising the use of the Morchella esculenta nutrition bag of claim 4 in a Morchella esculenta cultivation process. Preferably, the substances in the morchella nutrient bag are fully contacted with the soil of a morchella cultivation field.
10. A method for cultivating and producing seeds of morchella esculenta, which comprises the step of cultivating and producing seeds of the morchella esculenta by using any one of the culture mediums or the combination thereof as claimed in any one of claims 1 to 4, or any one of the cultivation, inoculation and/or use methods.
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