CN115777441A - Method for improving yield per unit of cordyceps militaris sporocarp - Google Patents
Method for improving yield per unit of cordyceps militaris sporocarp Download PDFInfo
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- CN115777441A CN115777441A CN202211422010.5A CN202211422010A CN115777441A CN 115777441 A CN115777441 A CN 115777441A CN 202211422010 A CN202211422010 A CN 202211422010A CN 115777441 A CN115777441 A CN 115777441A
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A40/00—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production
- Y02A40/10—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production in agriculture
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Abstract
The invention belongs to the technical field of artificial cultivation of cordyceps militaris and provides a method for improving the yield per unit of cordyceps militaris sporocarp. The method comprises the following steps: (1) Crushing grains, mixing with water, high temperature resistant alpha-amylase and pullulanase, soaking for 30-60 min, and then sequentially cooking and sterilizing to obtain a mixture; (2) Stirring the mixture while the mixture is hot, sequentially sieving and eluting, and then cooling to room temperature to obtain a grain culture medium; (3) Inoculating cordyceps militaris strains into a grain culture medium, supplementing water, and sequentially carrying out dark culture and illumination culture; and (4) harvesting after 50-60 days after inoculation. The invention removes the adhesive layer released in the process of cooking and sterilizing the grains, and fundamentally solves the problem of overlarge viscosity of the grain culture medium. The density of the cordyceps militaris fruiting body obtained by the method is obviously improved, the yield per unit of the cordyceps militaris is obviously improved, the improvement rate reaches more than 40%, and the economic benefit is obvious.
Description
Technical Field
The invention relates to the technical field of artificial cultivation of cordyceps militaris, in particular to a method for improving the yield per unit of cordyceps militaris sporocarp.
Background
Cordyceps militaris (Cordyceps militaris), also called Cordyceps militaris (L.) Link and Cordyceps militaris (L.) Link, belongs to Ascomycota of Ascomycota, sordariomycetes of Chaetomycete, hypocrea of Hypocreales, cordycipitaceae of Cordyceps, and Cordyceps of Cordyceps, and is a model species of Cordyceps. In recent years, research shows that the chemical components and pharmacological actions of artificially cultured cordyceps militaris are similar to those of cordyceps sinensis, and the artificially cultured cordyceps militaris is a culturable artificial cordyceps militaris variety which is most hopeful to replace the cordyceps sinensis. According to estimation, the industrial output value of the cordyceps militaris is estimated to be more than 200 hundred million yuan in 2022.
At present, the large-scale production of cordyceps militaris adopts a grain culture medium for cultivation, and the main grain raw materials comprise wheat, rice, millet, high corn, soybean meal, oat and the like. However, the grain raw material is rich in starch, gelatinization occurs in the cooking and sterilization process when the grain raw material meets water, grain particles are adhered to each other after cooling, the air permeability is reduced, hyphae are difficult to permeate after cordyceps militaris is inoculated, the primordium is less germinated after light is subjected to color conversion, the grass density is low, and the utilization rate of the final grain culture medium is low.
The patent (publication No. CN 105985911A) discloses a method for reducing the viscosity of a cereal culture medium, which is characterized in that sterilization and hydration are independently completed in two stages in a material steaming tank, wherein the mass ratio of the cereal to water in the sterilization stage is 1-1, and the mass ratio of the cereal to water in the hydration stage is 1.5-2. Finally, the cereal culture medium has high water absorption, loose particles and no stickiness.
The patent (publication No. CN 102895685A) discloses a sterilization system of a culture medium, a sterilization method using the system and a culture method of cordyceps militaris.
The above patent is an innovation in the sterilization process by independently completing the sterilization and hydration of the substrate in two stages or adding a vacuum pumping operation after the sterilization. However, the inventors of the present invention found in actual production that neither sterilization in two stages nor evacuation operation after sterilization could fundamentally find out the cause of the increase in viscosity, i.e., the presence of the adhesion layer. During the cooking and sterilization process of grains, when the grains and water are heated and gelatinized, a large amount of free amylose and amylopectin are dissolved out, and gel is formed after cooling and is wrapped on the surfaces of the grains to form a water retention film, namely an adhesion layer. It is understood that the adhesion layer is a layer of pasty high-moisture substance covering the surface of the grain. Then, no matter two-stage sterilization or improvement of a vacuumizing process is carried out, the viscous component is not removed, so that the local viscosity of culture medium particles is not substantially reduced, and because of the existence of an adhesive layer, the phenomenon that cordyceps militaris hyphae contacts with the surface component of the grains to influence the utilization rate of the grains is avoided.
Disclosure of Invention
In order to overcome the defects in the prior art, the invention provides a method for improving the yield per unit of cordyceps militaris sporocarp, which can remove an adhesive layer released in the process of stewing and sterilizing grains, thereby fundamentally solving the problem of overlarge viscosity of a grain culture medium.
In order to achieve the above object, the present invention provides the following technical solutions:
the invention provides a method for improving the yield per unit of cordyceps militaris sporocarp, which comprises the following steps:
(1) Crushing grains, mixing with water, high temperature resistant alpha-amylase and pullulanase, soaking for 30-60 min, and then sequentially cooking and sterilizing to obtain a mixture;
(2) Stirring the mixture while the mixture is hot, sequentially sieving and eluting, and then cooling to room temperature to obtain a grain culture medium;
(3) Inoculating cordyceps militaris strains into a grain culture medium, supplementing water, and sequentially carrying out dark culture and illumination culture;
(4) Harvesting after 50-65 days after inoculation.
Preferably, the mesh number of the pulverization in the step (1) is-10 to +30 meshes.
Preferably, the grains in the step (1) comprise one or more of wheat, rice, brown rice, polished round-grained rice, barley, corn, oat, buckwheat and soybean meal, the mixing mass ratio of the grains to water is 1-2, the dosage of the high-temperature resistant alpha-amylase is 0.01-0.1% of the mass of the grains, and the dosage of the pullulanase is 0.005-0.2% of the mass of the grains.
Preferably, the cooking temperature in the step (1) is 65-100 ℃, and the cooking time is 20-40 min.
Preferably, the temperature for sterilization in the step (1) is 120-130 ℃, and the time for sterilization is 20-40 min.
Preferably, the mesh number of the sieving in the step (2) is-10 to +30 meshes.
Preferably, the elution in the step (2) is to elute an adhesion layer formed on the surface of the grain, and the elution amount of the adhesion layer is 10-30% of the mass of the grain.
Preferably, the inoculation amount of the cordyceps militaris strain in the step (3) is 5-10% of the mass of the grains, and the supplemented water amount is 10-30% of the mass of the grains.
Preferably, the temperature of the dark culture in the step (3) is 16-19 ℃, and the time of the dark culture is 3-7 days.
Preferably, the illumination intensity of the illumination culture in the step (3) is 50 to 1000Lux, and the temperature of the illumination culture is 16 to 22 ℃.
Compared with the prior art, the invention has the following beneficial effects:
1. the invention removes the adhesive layer released in the process of cooking and sterilizing the grains, and fundamentally solves the problem of overlarge viscosity of the grain culture medium.
2. The invention provides a method for carrying out enzymolysis by using high-temperature resistant alpha-amylase and pullulanase, carrying out enzymolysis damage on a dissolved gel structure, promoting a large amount of dissolution of an adhesive layer, fully degrading amylose and amylopectin, and ensuring the smoothness of elution and filtration. The high temperature resistant alpha-amylase and the pullulanase can also form holes with different sizes on the surface of the grains, thereby improving the specific surface area of the grains and being beneficial to further utilization of the cordyceps militaris mycelia.
3. After the adhesion layer is removed, the grain culture medium is changed from the original compact state to the loose state, the porosity among grains is increased, the nutrition on the grain surface can be fully exposed around the cordyceps militaris hypha, and the oxygen supply is sufficient, so that the hypha grows robustly, and the utilization rate of the grain is greatly improved.
4. The eluted adhesive layer can be used for preparing cordyceps militaris liquid strains and also can be used as a high-quality raw material for producing glucose, so that the additional value of cordyceps militaris production is improved.
5. The density of the cordyceps militaris fruiting body obtained by the method is obviously improved, the yield per unit of the cordyceps militaris is obviously improved, the improvement rate reaches more than 40%, and the economic benefit is obvious.
Drawings
In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings used in the description of the embodiments or the prior art will be briefly described below, it is obvious that the drawings in the following description are only embodiments of the present invention, and for those skilled in the art, other drawings can be obtained according to the provided drawings without creative efforts.
FIG. 1 shows the state of the medium in comparative example 2 and example 2 (note: the left panel shows comparative example 2, and the right panel shows example 2);
FIG. 2 shows the spawn running state of comparative example 2 and example 2 (note: the left figure shows comparative example 2, and the right figure shows example 2);
FIG. 3 shows the middle growth state of Cordyceps militaris in comparative example 2 and example 2 (note: the left figure is comparative example 2, and the right figure is example 2);
FIG. 4 shows the late growth state of Cordyceps militaris in comparative example 2 and example 2 (note: the left image is comparative example 2, and the right image is example 2).
Detailed Description
The invention provides a method for improving the yield per unit of cordyceps militaris sporocarp, which comprises the following steps:
(1) Crushing grains, mixing with water, high-temperature-resistant alpha-amylase and pullulanase, soaking for 30-60 min, and then sequentially cooking and sterilizing to obtain a mixture;
(2) Stirring the mixture while the mixture is hot, sequentially sieving and eluting, and then cooling to room temperature to obtain a grain culture medium;
(3) Inoculating cordyceps militaris strains into a grain culture medium, supplementing water, and sequentially carrying out dark culture and illumination culture;
(4) Collected 50-65 days after inoculation.
In the invention, the cereal is crushed in the step (1), mixed with water, high temperature resistant alpha-amylase and pullulanase, soaked for 30-60 min, and then sequentially cooked and sterilized to obtain a mixture, and more preferably, the cereal is crushed, mixed with water, high temperature resistant alpha-amylase and pullulanase, soaked for 45min, and then sequentially cooked and sterilized to obtain a mixture.
In the present invention, the mesh size of the pulverization in the step (1) is preferably-10 to +30 mesh, more preferably-15 to +25 mesh, and still more preferably-15 to +20 mesh, wherein "-" indicates a mesh size that can pass through the mesh size, and "+" indicates a mesh size that cannot pass through the mesh size.
In the invention, the grains in the step (1) preferably comprise one or more of wheat, rice, brown rice, polished round-grained rice, barley, corn, oat, buckwheat and soybean meal, and are further preferably oat; the mixing mass ratio of the grains to the water is preferably 1; the dosage of the high-temperature resistant alpha-amylase is preferably 0.01 to 0.1 percent of the mass of the grains, more preferably 0.01 to 0.1 percent of the mass of the grains, and even more preferably 0.04 percent of the mass of the grains; the pullulanase is preferably used in an amount of 0.005 to 0.2% by mass of the grain, more preferably 0.005 to 0.08% by mass of the grain, and still more preferably 0.005% by mass of the grain.
In the present invention, the temperature of the cooking in step (1) is preferably 65 to 100 ℃, more preferably 70 to 90 ℃, and still more preferably 80 ℃; the cooking time is preferably 20 to 40min, more preferably 25 to 35min, and still more preferably 30min.
In the present invention, the temperature for the sterilization in the step (1) is preferably 120 to 130 ℃, more preferably 121 to 123 ℃, and still more preferably 122 ℃; the time for sterilization is preferably 20 to 40min, more preferably 25 to 35min, and still more preferably 30min.
In the present invention, the mesh size of the screen in step (2) is preferably-10 to +30 mesh, more preferably-15 to +25 mesh, and still more preferably-15 to +20 mesh, wherein "-" indicates a mesh size that can pass through the mesh size, and "+" indicates a mesh size that cannot pass through the mesh size.
In the present invention, the elution in step (2) is preferably to elute an adhesion layer formed on the surface of the grain, and the elution amount of the adhesion layer is preferably 10 to 30% by mass of the grain, more preferably 15 to 29.5% by mass of the grain, and still more preferably 18% by mass of the grain.
In the invention, the inoculation amount of the cordyceps militaris strain in the step (3) is preferably 5-10% of the mass of the grain, more preferably 7-9% of the mass of the grain, and even more preferably 8% of the mass of the grain; the amount of the supplementary water is preferably 10 to 30% by mass of the cereal, more preferably 12 to 25% by mass of the cereal, and still more preferably 15% by mass of the cereal.
In the present invention, the temperature of the dark culture in the step (3) is preferably 16 to 19 ℃, and more preferably 18 ℃; the dark culture time is preferably 3 to 7 days, more preferably 4 to 6 days, and still more preferably 5 days.
In the present invention, the light intensity of the light culture in the step (3) is preferably 50 to 1000Lux, more preferably 200 to 600Lux, and still more preferably 500Lux; the temperature of the light culture is preferably 16 to 22 ℃, more preferably 19 to 21 ℃, and still more preferably 20 ℃.
In the present invention, the harvest is performed 50 to 65 days after the inoculation in step (4), and more preferably the harvest is performed 60 days after the inoculation.
The technical solutions provided by the present invention are described in detail below with reference to examples, but they should not be construed as limiting the scope of the present invention.
The liquid Cordyceps militaris strains in the following examples and comparative examples are cultured in PDB-enriched medium (the PDB-enriched medium uses water as solvent and comprises the following components with the following concentrations of 200g/L of potato, 10g/L of silkworm chrysalis meal, 10g/L of glucose, 2g/L of peptone and KH 2 PO 4 0.5g/L，MgSO 4 1g/L, pH adjusted to 7.0 with sodium hydroxide) and performing conventional shake culture (20 deg.C, 130rmp,7 days) to obtain the mother strain from Honghao biological technology limited of Jiangmen city.
Example 1
(1) And (3) cooking of wheat:
sieving wheat to remove impurities, mixing the crushed and trapped wheat particles between-10 meshes and +30 meshes with water, high-temperature resistant alpha-amylase and pullulanase, soaking at normal temperature for 30min, and then cooking, wherein the mass ratio of the wheat to the water is 1. The cooking temperature is set to 65 ℃ and the cooking time is set to 40min.
(2) And (3) sterilization: steaming the crushed wheat and water, heating the mixture to 121 ℃, and sterilizing for 30min; the cultivation bottles are independently subjected to damp-heat sterilization at 121 ℃ for 30min.
(3) And (3) elution: stirring thoroughly while hot, washing with sterile water containing 50% of crushed wheat, sieving (-10 to +30 mesh) and eluting sequentially, wherein the elution amount of the adhesion layer is 29.5% of that of the grain.
(4) Inoculation: after the eluted broken wheat grains are cooled, cordyceps militaris liquid strains are uniformly sprayed into a broken wheat culture medium, and the inoculation amount is 5% of the quality of the broken wheat.
(5) Filling: under the aseptic condition, an automatic racking machine fills the inoculated cordyceps militaris culture medium into a cultivation bottle, additionally adds sterile water with the mass of 25% of broken wheat, fills the bottle with the amount of 290 g/bottle, and covers the bottle.
(6) And (3) cultivating: placing the Cordyceps militaris culture medium into a culture room for dark culture, and culturing at 16 deg.C for 3 days until the Cordyceps militaris culture medium surface is completely covered by Cordyceps militaris mycelia; after completion of the dark culture, 50Lux light was applied and cultivation management was performed at 16 ℃.
(7) Harvesting: harvested on day 55 after inoculation.
Example 2
(1) Cooking oat:
sieving oats to remove impurities, mixing the crushed and trapped oat particles with water, high-temperature-resistant alpha-amylase and pullulanase, soaking the mixture for 45min at normal temperature, and then cooking the mixture, wherein the mass ratio of the oats to the water is 1. The cooking temperature is set to 80 ℃, and the cooking time is set to 30min.
(2) And (3) sterilization: continuously heating the mixture of oat and water after cooking to 122 ℃, and sterilizing for 30min; the cultivation bottles are independently subjected to damp-heat sterilization at the temperature of 122 ℃ for 30min.
(3) And (3) elution: and after fully stirring the mixture while the mixture is hot, sequentially sieving the mixture to (-15 to +20 meshes) and eluting the mixture, wherein the elution amount of an adhesive layer accounts for 18 percent of that of the grains.
(4) Inoculation: after the eluted oat particles are cooled, the cordyceps militaris liquid strain is uniformly sprayed into an oat culture medium, and the inoculation amount is 8% of the mass of the oat.
(5) Filling: under the aseptic condition, an automatic racking machine fills the inoculated cordyceps militaris culture medium into a culture bottle, sterile water with the mass of 15% of oat is additionally added, the filling amount is 290 g/bottle, and a cover is covered.
(6) Culturing: placing the Cordyceps militaris culture medium into a culture room for dark culture, and culturing at 18 deg.C for 5 days until the Cordyceps militaris culture medium surface is completely covered by Cordyceps militaris mycelia; after completion of the dark culture, 500Lux light was applied and cultivation management was performed at 20 ℃.
(7) Harvesting: collected 60 days after inoculation.
Example 3
(1) And (3) steaming and boiling brown rice:
the brown rice is sieved to remove impurities, the brown rice particles which are crushed and cut between minus 10 meshes to plus 30 meshes are mixed with water, high-temperature resistant alpha-amylase and pullulanase, the mixture is soaked for 60min at normal temperature and then is steamed, the mass ratio of the brown rice to the water is 1. The cooking temperature is set to 65 ℃ and the cooking time is set to 20min.
(2) And (3) sterilization: after the brown rice and the water are cooked, the mixture is continuously heated to 128 ℃, and the sterilization time is 30min; the cultivation bottles are independently subjected to damp-heat sterilization at the sterilization temperature of 123 ℃ for 30min.
(3) And (3) elution: and after fully stirring the mixture while the mixture is hot, sequentially sieving the mixture to (-10 to +30 meshes) and eluting the mixture, wherein the elution amount of the adhesion layer accounts for 25.6 percent of that of the grains.
(4) Inoculation: after the eluted brown rice particles are cooled, cordyceps militaris liquid strains are uniformly sprayed into a brown rice culture medium, and the inoculation amount is 10% of the mass of the brown rice.
(5) Filling: under the aseptic condition, the inoculated cordyceps militaris culture medium is filled into a cultivation bottle by an automatic racking machine, sterile water with the mass of 20% of that of the brown rice is additionally added, the filling amount is 290 g/bottle, and a cover is covered.
(6) Culturing: placing the Cordyceps militaris culture medium into a culture room for dark culture, and culturing at 19 deg.C for 7 days until the Cordyceps militaris culture medium surface is completely covered by Cordyceps militaris mycelia; after completion of the dark culture, 1000Lux light was applied and cultivation management was performed at 22 ℃.
(7) Harvesting: harvested 65 days after inoculation.
Comparative example 1
The only difference from example 1 is that the grain was not subjected to the milling and elution steps, and the remaining steps and step parameters were consistent with those of example 1.
Comparative example 2
The only difference from example 2 is that the grain was not subjected to the milling and elution steps, and the remaining steps and step parameters were consistent with those of example 2.
Comparative example 3
The only difference from example 3 is that the grains were not subjected to the milling and elution steps, and the remaining steps and parameters of the steps were the same as those of example 3.
Experimental example 1
The results of comparing the yields per unit of the cordyceps militaris fruit bodies obtained in examples 1 to 3 and comparative examples 1 to 3 are shown in table 1.
TABLE 1 Individual yield of fruiting bodies of Cordyceps militaris obtained by different treatments
As can be seen from Table 1, the method of examples 1 to 3 of the present invention significantly improves the yield per unit of fruiting body of Cordyceps militaris, as compared with comparative examples 1 to 3.
In conclusion, the invention adopts the grains as the culture medium, removes a large amount of adhesive layers released by the grains in the cooking and sterilizing process through cooking and sterilizing, eluting and filtering, and fundamentally solves the common problem of high viscosity of the grain culture medium in the industry. Firstly, due to the removal of the adhesion layer, the grain culture medium becomes loose, the porosity among the grains is increased, the supply of oxygen is enhanced, the cordyceps militaris mycelia can rapidly penetrate into the grain gaps, and the nutrition on the surface of the grains is directly utilized. And secondly, the high-temperature resistant alpha-amylase and the pullulanase can also form holes with different sizes on the surface of the grains, so that the specific surface area of the grains is improved, and the further utilization of cordyceps militaris mycelia is facilitated. Finally, the eluted adhesive layer can be used for preparing cordyceps militaris liquid seeds and can also be used as a high-quality raw material for producing glucose, so that the additional value of cordyceps militaris production is improved. Overall, the method of the invention has the following beneficial effects: the mycelium grows vigorously, the grass density of the cordyceps militaris is obviously improved, and the yield per unit of the cordyceps militaris is improved by over 40 percent. The method has obvious economic benefit and is worthy of large-scale popularization and application.
The foregoing is only a preferred embodiment of the present invention, and it should be noted that, for those skilled in the art, various modifications and decorations can be made without departing from the principle of the present invention, and these modifications and decorations should also be regarded as the protection scope of the present invention.
Claims (10)
1. A method for improving the yield per unit of cordyceps militaris sporocarp is characterized by comprising the following steps:
(1) Crushing grains, mixing with water, high temperature resistant alpha-amylase and pullulanase, soaking for 30-60 min, and then sequentially cooking and sterilizing to obtain a mixture;
(2) Stirring the mixture while the mixture is hot, sequentially sieving and eluting, and then cooling to room temperature to obtain a grain culture medium;
(3) Inoculating cordyceps militaris strains into a grain culture medium, supplementing water, and sequentially carrying out dark culture and illumination culture;
(4) Collected 50-65 days after inoculation.
2. The method for improving the yield of the fruiting body of cordyceps militaris according to claim 1, wherein the crushed meshes in the step (1) are-10 to +30 meshes.
3. The method for improving the yield of the fruiting body of cordyceps militaris according to claim 1 or 2, wherein the grain in the step (1) comprises one or more of wheat, rice, brown rice, polished round-grained rice, barley, corn, oat, buckwheat and soybean meal, the mixing mass ratio of the grain to water is 1.
4. The method for improving the yield per unit of fruiting body of Cordyceps militaris as claimed in claim 3, wherein the cooking temperature in step (1) is 65-100 deg.C, and the cooking time is 20-40 min.
5. The method for increasing the yield of the fruiting body of Cordyceps militaris as claimed in claim 4, wherein the temperature of sterilization in step (1) is 120-130 ℃, and the time of sterilization is 20-40 min.
6. The method for improving the yield of the fruiting body of Cordyceps militaris as claimed in claim 5, wherein the mesh number of the sieving in step (2) is-10 to +30 meshes.
7. The method for improving the yield of the fruiting body of cordyceps militaris according to claim 6, wherein the elution in the step (2) is to elute an adhesion layer formed on the surface of the grain, and the elution amount of the adhesion layer is 10-30% of the mass of the grain.
8. The method for improving the yield of the fruiting body of cordyceps militaris according to claim 7, wherein the inoculation amount of the cordyceps militaris strain in the step (3) is 5-10% of the mass of the grain, and the amount of the supplemented water is 10-30% of the mass of the grain.
9. The method for increasing the yield of the fruiting body of Cordyceps militaris as claimed in claim 8, wherein the temperature of dark culture in step (3) is 16-19 ℃ and the time of dark culture is 3-7 days.
10. The method for increasing the yield of the fruiting body of cordyceps militaris according to claim 9, wherein the illumination intensity of the illumination culture in the step (3) is 50-1000 Lux, and the temperature of the illumination culture is 16-22 ℃.
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| JP2017046687A (en) * | 2015-09-01 | 2017-03-09 | 学校法人甲南学園 | Mushroom culture medium liquids, mushroom culture media, and production methods of mushroom culture medium liquids, and mushroom cultivation methods |
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| CN109312371A (en) * | 2016-04-04 | 2019-02-05 | 由农业部部长代表的美利坚合众国 | The method for handling cereal |
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| JP2017046687A (en) * | 2015-09-01 | 2017-03-09 | 学校法人甲南学園 | Mushroom culture medium liquids, mushroom culture media, and production methods of mushroom culture medium liquids, and mushroom cultivation methods |
| CN109312371A (en) * | 2016-04-04 | 2019-02-05 | 由农业部部长代表的美利坚合众国 | The method for handling cereal |
| CN108207491A (en) * | 2018-01-13 | 2018-06-29 | 蚌埠学院 | A kind of method using wheat embryo cultivation fruiting bodies of cordyceps militaris |
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