A culture medium for chrysophryllum aurantiacum and a preparation method thereof

Abstract

The application discloses a culture medium for Floccularia luteovirens and a preparation method thereof, and belongs to the technical field of edible fungus cultivation. The culture medium is used for artificial domestication culture of a Floccularia luteovirens strain with a preservation number of GDMCC66843 and a name of Floccularia luteovirens S4-1, and comprises mother culture medium, liquid fermentation culture medium and cultivation medium. The liquid fermentation culture medium comprises the following components in mass fraction: 30-40 g of wheat powder, 1-3 g of yeast extract, 0.5-1.5 g of potassium dihydrogen phosphate and 3-5 g of temperature conducting agent are added into one liter of solvent; the solvent is water; the target temperature is 25 / 10 DEG C; the temperature conducting agent is a pH-responsive heat-conducting modified fiber; and the temperature is alternately changed every 12 h for 20 d. The application is suitable for temperature changing fermentation culture, not only realizes propagation of the strain, and the obtained cultivation strain can be successfully cultivated in the cultivation medium.

Description

TECHNICAL FIELD

[0001] The application belongs to the technical field of edible mushroom cultivation and relates to a Floccularia luteovirens culture medium and a preparation method thereof. BACKGROUND

[0002] Floccularia luteovirens, also known as butter yellow mushroom, grassland yellow mushroom and Shiqu white mushroom, is a wild edible mushroom unique to the Qinghai-Tibet Plateau region. It mainly grows in alpine meadows at an altitude of 3000-5000 m and is distributed in Qinghai Province, Tibet Autonomous Region, Sichuan Province and Gansu Province of China. The prior art lacks a culture medium for artificial cultivation of Floccularia luteovirens. SUMMARY

[0003] The application aims to provide a Floccularia luteovirens culture medium and a preparation method thereof, and solve the problem that the prior art lacks a culture medium for artificial cultivation of Floccularia luteovirens.

[0004] The technical scheme adopted by the application is as follows.

[0005] A Floccularia luteovirens culture medium is used for artificial domestication culture of a Floccularia luteovirens strain with a preservation number of GDMCC66843 and a name of Floccularia luteovirens S4-1, and includes a mother strain culture medium, a liquid fermentation culture medium and a cultivation medium.

[0006] The liquid fermentation culture medium includes the following components in mass fraction: 30-40 g of wheat powder, 1-3 g of yeast extract, 0.5-1.5 g of potassium dihydrogen phosphate and 3-5 g of a temperature conducting agent are added to each liter of solvent, the solvent is water, and the culture medium is suitable for 20 days of alternate temperature culture every 12 hours between 25 / 10 DEG C.

[0007] The temperature conducting agent is a pH-responsive heat-conducting modified fiber and includes the following components: an acid-sensitive liposome containing a cellulase dispersion liquid inside, boron nitride nanosheet modified protein-cellulose fiber and low-viscosity starch modified by low-temperature anticoagulation.

[0008] The acid-sensitive liposome is loaded on the boron nitride nanosheet modified protein-cellulose fiber impregnated with low-viscosity starch.

[0009] The Floccularia luteovirens has a large diurnal temperature difference in the original environment, and the conventional constant temperature culture cannot promote the proliferation of mycelium and the success rate of cultivation after proliferation; therefore, in the research process, the application introduces variable temperature liquid fermentation deep culture with a temperature difference of about 15 DEG C, and the change of the temperature difference can promote the proliferation of mycelium and strengthen the growth force of the mycelium in the cultivation process.

[0010] The liquid fermentation culture medium in the application is alternately cultured at 25 / 10 DEG C every 12 hours, in the early stage of culture, the mycelium is dispersed and small in amount, when the temperature changes, the mycelium cannot form a continuous temperature conduction path between the mycelium, and the environment around the mycelium growth is prone to uneven temperature change, uneven heating of the mycelium, etc., which affects the propagation curve of the mycelium, and cannot form a stable upward change curve, affecting the quality of the cultivated species.

[0011] Therefore, the application introduces a fibrous temperature conduction agent, the fibrous structure is similar to the mycelium structure in macroscopic view, the application supplements the fibrous material with certain heat conduction effect in the early stage of fermentation, which makes up for the defects of small amount and dispersion of the mycelium, the fibrous material is inserted between the mycelium, and a heat conduction network structure is artificially constructed, which can uniformly and smoothly change the temperature during heating and cooling, and realize smooth conversion of the temperature; avoid uneven heating of the mycelium and rapid change of the temperature from high to low or from low to high in a short time, which leads to insufficient temperature change adaptation time.

[0012] With the growth and reproduction of the mycelium, the mycelium has strong environmental adaptation ability, and a large amount of mycelium needs to interact and intertwine to form a relatively dense mycelial ball, which simplifies the solid-liquid separation in the later stage, and the mycelial ball is deposited at the bottom of the container to realize solid-liquid separation, in order to avoid the mycelial ball mixed with the fibrous material or the fibrous material affecting the growth and reproduction of the mycelium due to space occupation, the heat conduction modified fiber in the application is mainly composed of protein and cellulose, the activity of extracellular enzyme is strengthened with the growth and reproduction of the mycelium, enzyme hydrolysis occurs, and the hydrolysis product is utilized by the mycelium to promote the growth and reproduction of the mycelium; after the protein and cellulose are hydrolyzed, the fiber structure is disintegrated, which does not affect the growth and reproduction of the mycelium, and does not increase the difficulty of solid-liquid separation of the fermentation system. Boron nitride nanosheet is an inert material and has good heat conductivity, which can significantly improve the heat conductivity of the fiber, appropriately improve the temperature change speed, and the boron nitride nanosheet is an inert material and does not react with the mycelium or other substances in the fermentation culture medium, and does not have a competitive relationship with the mycelium, and does not hinder the growth and reproduction of the mycelium, the application uses pure boron nitride nanosheet, which is transparent, even if it is dispersed in the upper liquid, it is also transparent, and the nanomaterial is difficult to be seen by the naked eye, which does not affect the observation of the fermentation condition in the container by artificial.

[0013] In order to improve the winding continuity between fibers and fibers, fibers and mycelium, and form a network heat conduction structure with good continuity, the application adopts low-viscosity starch with low-temperature anti-coagulation modification to modify the boron nitride nanosheet modified protein-cellulose fiber for impregnation. The low-viscosity starch can form a sticky layer around the fiber with higher viscosity than water but without causing physical obstruction to the growth of mycelium. After mixing the fibers and mycelium evenly, the sticky layer can effectively prevent the large-scale separation of intertwined fibers and fibers, and fibers and mycelium, which helps to maintain the structural stability of the whole heat conduction system in the early stage of mycelium growth and reproduction. With the improvement of amylase activity in the metabolic process of mycelium, starch is hydrolyzed, and in the later stage of mycelium growth and reproduction, it will not hinder the solid-liquid separation of the whole system, nor will it cause the mycelium ball to be too dense.

[0014] The activity of cellulase produced by mycelium metabolism in the application is high in the middle and later stages of culture, but the main component of the fiber is cellulose. In order to realize the complete hydrolysis of the fiber as much as possible, the application supplements acid-sensitive liposomes containing cellulase dispersion. The whole fermentation system becomes acidic with the decrease of pH during fermentation. The application takes advantage of this natural change to use acid-sensitive liposomes to encapsulate cellulase. Under certain acidic conditions, acid-sensitive liposomes begin to crack and release the internal cellulase, which cooperates with the cellulase in the metabolites to promote the hydrolysis of the fiber, so that the fiber can be completely disintegrated as much as possible when the fermentation culture reaches the end point. A small amount of unhydrolyzed material will be evenly dispersed in the upper liquid. The complete hydrolysis of the fiber as much as possible can provide more nutrient substances for the mycelium on the one hand, and reduce the spatial obstruction of the fiber to the growth and reproduction of the mycelium on the other hand.

[0015] In summary, the introduction of heat-conducting fibers in the application reduces the influence of temperature change on mycelium in the early stage of mycelium fermentation culture. With the hydrolysis of a large number of components in the heat-conducting fiber, the adverse effects of the fiber on the growth and reproduction of the mycelium in the later stage are avoided. At the same time, more nutrient substances are continuously provided for the mycelium in the hydrolysis process of the fiber components, promoting the growth and reproduction of the mycelium.

[0016] Further, the low-viscosity starch with low-temperature anti-coagulation modification includes low-viscosity tartaric acid starch ester and poly-N-isopropyl acrylamide grafted starch complex, and the mass ratio of the low-viscosity tartaric acid starch ester and the poly-N-isopropyl acrylamide grafted starch is 3:1.

[0017] Further, the boron nitride nanosheet modified protein-cellulose fiber includes the following components by weight: 50 parts of ionic liquid, 20-25 parts of bamboo cellulose, 5-10 parts of protein, 0.5-1 part of lactic acid oligomer, and 3-5 parts of boron nitride nanosheet.

[0018] The method comprises the following steps: adding boron nitride nanosheets into an ionic liquid, adding bamboo cellulose and lactic acid oligomers after heating reaction, then adding a protein suspension, uniformly mixing to obtain a blended spinning solution, and then performing spinning and cross-linking to obtain boron nitride nanosheet modified protein-cellulose fibers.

[0019] Further, the protein comprises at least one of soybean protein, rice protein, peanut protein and rapeseed protein.

[0020] Further, the cellulase in the temperature-conducting agent accounts for 1-1.5% of the mass of the boron nitride nanosheet modified protein-cellulose fiber, and the response pH of the acid-sensitive liposome is less than 5.5.

[0021] Further, the mother culture medium comprises PDA culture medium, sawdust powder, VB2 and growth promoting compound, the amount of the sawdust powder accounts for 30% of the mass of the PDA culture medium, the amount of the VB2 accounts for 0.1% of the mass of the PDA culture medium, and the amount of the growth promoting compound accounts for 20% of the mass of the PDA culture medium.

[0022] Further, the growth promoting compound comprises wheat powder, ammonium sulfate, magnesium sulfate and potassium chloride, and the mass ratio of the wheat powder, the ammonium sulfate, the magnesium sulfate and the potassium chloride is 5:1:0.2:0.2.

[0023] Further, the cultivation medium comprises solid material and water, and the ratio of the solid material to the water is 1:1.6.

[0024] The solid material comprises the following components in terms of mass fraction: 98% of amino acid compound-main compound, 1% of superphosphate and 1% of gypsum.

[0025] The main compound in the cultivation medium comprises sawdust powder, cottonseed hull, corn cob, bran, corn powder and soybean meal, and the mass ratio of the sawdust powder, the cottonseed hull, the corn cob, the bran, the corn powder and the soybean meal is 6:5:3.6:3:1:1.

[0026] The amino acid compound in the cultivation medium comprises arginine, proline, histidine and lysine, and the mass ratio of the arginine, the proline, the histidine and the lysine is 3:1:1:1.

[0027] The amino acid compound-main compound is obtained by physically mixing the amino acid compound and the main compound in a mass ratio of 1:4.

[0028] Further, the initial pH of the liquid fermentation medium is 6.0, and the inoculation amount of the liquid fermentation medium is 7% of the volume of the liquid fermentation medium.

[0029] A preparation method of a yellow-green volva mushroom culture medium, comprising the following steps:

[0030] S1, wood chip powder, VB2 and growth promoting compound are sequentially added in PDA culture medium to prepare a mother culture medium, the mother culture is multiplied to obtain a primary culture;

[0031] S2, after water is added in a container, wheat powder, yeast extract, potassium dihydrogen phosphate and temperature conducting agent are sequentially added and uniformly dispersed to obtain a uniform suspension, the suspension is a liquid fermentation culture medium, the primary culture in step S1 is inoculated in the liquid fermentation culture medium for further multiplication to obtain a cultivation culture;

[0032] S3, amino acid compound-main compound is used as main material, supplemented with calcium superphosphate and gypsum, and then mixed uniformly, water is added and stirred uniformly according to a material-water ratio of 1:1.6 to obtain a cultivation medium for inoculating the cultivation culture.

[0033] As described above, the present application has the following advantages:

[0034] 1. The yellow-green volva mushroom culture medium comprises a mother culture medium, a liquid fermentation culture medium and a cultivation medium, the liquid fermentation culture medium is used as an intermediate bridge to multiply the culture, and the culture is obtained by variable temperature fermentation, so that the culture is multiplied, and the cultivation culture can be successfully cultivated in the cultivation medium;

[0035] 2. The temperature conducting agent in a fibrous form is introduced, the fibrous structure is similar to the mycelium structure in macroscopic view, the fibrous form with a certain heat conducting effect is supplemented in the early fermentation stage to compensate for the defects of less mycelium and dispersion, the fibers are inserted between the mycelium, a heat conducting network structure is artificially constructed, the temperature can be uniformly and stably changed during temperature rising and falling, and smooth temperature conversion is realized; the mycelium is not uniformly heated, and the temperature is changed from high to low or from low to high in a short time, so that there is not enough temperature change adaptation time;

[0036] 3. The influence of temperature change on the mycelium in the early mycelium fermentation culture is reduced by introducing the heat conducting fibers, the adverse effects of the fibers on the growth and reproduction of the mycelium in the later stage are avoided, and more nutrients are continuously provided for the mycelium in the hydrolysis process of the fiber components, so that the growth and reproduction of the mycelium are promoted. BRIEF DESCRIPTION OF DRAWINGS

[0037] To more clearly illustrate the technical solutions of the embodiments of the present invention, the accompanying drawings used in the embodiments will be briefly described below. It should be understood that the following drawings only show some embodiments of the present invention and should not be regarded as a limitation of the scope. For those skilled in the art, other related drawings can be obtained based on these drawings without creative effort, wherein:

[0038] Figures 1-3 This describes the mycelial proliferation after passing through a liquid fermentation medium at different temperatures;

[0039] Figure 4 This describes the process of temperature-controlled cultivation according to the present invention.

[0040] Figure 5 This is a curve of mycelial growth in variable temperature culture according to the present invention;

[0041] Figures 6-7 These are images showing the mycelial growth and actual specimens under different material-to-water ratios in the cultivation substrate of this invention.

[0042] Figure 8 This is a diagram of the white, fluffy structure of the microorganisms after germination in the cultivation substrate in Example 2.

[0043] Figure 9 The graph shows the effect of culture time on the extracellular enzyme activity of liquid cultivars of *Fragaria leucocephala* in Example 2. Detailed Implementation

[0044] To make the objectives, technical solutions, and advantages of this invention clearer, the invention will be further described in detail below with reference to the accompanying drawings and embodiments. It should be understood that the specific embodiments described herein are only for explaining the invention and are not intended to limit the invention; that is, the described embodiments are merely some embodiments of the invention, and not all embodiments. The components of the embodiments of the invention described and shown in the accompanying drawings can generally be arranged and designed in various different configurations.

[0045] Therefore, the following detailed description of the embodiments of the invention provided in the accompanying drawings is not intended to limit the scope of the claimed invention, but merely to illustrate selected embodiments of the invention. All other embodiments obtained by those skilled in the art based on the embodiments of the invention without inventive effort are within the scope of protection of the invention.

[0046] It should be noted that the relational terms herein such as first and second and the like are used solely to distinguish one from another entity or action without necessarily requiring or implying any actual relationship or order between such entities or actions. Moreover, the terms "comprises", "comprising", or any other variations thereof, are intended to cover a non-exclusive inclusion, such that a process, method, article, or apparatus that comprises a list of elements does not include only those elements but can also include other elements not expressly listed or inherent to such process, method, article, or apparatus. An element proceeded by "comprises... a" does not, without more constraints, exclude the existence of additional identical elements in the process, method, article, or apparatus that comprises the element.

[0047] The features and properties of the present application will be further described with reference to the following examples.

[0048] The artificial domestication culture of the Floccularia luteovirens S4-1 mushroom strain with the preservation number GDMCC66843 and the name Floccularia luteovirens S4-1 provided by the embodiment of the present application includes mother culture medium, liquid fermentation culture medium and cultivation medium; the Floccularia luteovirens S4-1 mushroom strain with the preservation number GDMCC66843 is the mother culture, the preservation time is 2025.08.14, and the preservation address is the 5th floor of No. 59 Building, the Institute of Microbiology of Guangdong Academy of Sciences, 100, Martyrs' Road, Guangzhou;

[0049] The liquid fermentation culture medium includes the following components with mass fractions: 30-40 g of wheat flour, 1-3 g of yeast extract, 0.5-1.5 g of potassium dihydrogen phosphate and 3-5 g of temperature conductive agent are added per liter of solvent, the solvent is water, and the culture is suitable for alternating temperature culture between 25 / 10℃ every 12 h for 20 d;

[0050] The temperature conductive agent is a pH-responsive heat-conductive modified fiber, which includes the following components: acid-sensitive liposomes containing cellulase dispersion liquid, boron nitride nanosheet modified protein-cellulose fiber, low-viscosity starch modified by low-temperature anticoagulation;

[0051] The acid-sensitive liposomes are loaded on the boron nitride nanosheet modified protein-cellulose fiber impregnated with low-viscosity starch.

[0052] The low-viscosity starch modified by low-temperature anticoagulation includes low-viscosity tartaric acid ester of starch and poly-N-isopropyl acrylamide grafted starch complex, and the mass ratio of the low-viscosity tartaric acid ester of starch and the poly-N-isopropyl acrylamide grafted starch is 3:1.

[0053] The boron nitride nanosheet modified protein-cellulose fiber comprises the following components in parts by weight: 50 parts of ionic liquid, 20-25 parts of bamboo cellulose, 5-10 parts of protein, 0.5-1 part of lactic acid oligomer, and 3-5 parts of boron nitride nanosheet; wherein the ionic liquid is an alkylimidazole type ionic liquid; and the lactic acid oligomer is a low molecular weight polylactic acid with a molecular weight of 2000.

[0054] The boron nitride nanosheet modified protein-cellulose fiber is prepared by the following method: adding boron nitride nanosheet in ionic liquid, heating and reacting, then adding bamboo cellulose and lactic acid oligomer for further heating and reaction, subsequently adding protein suspension after cooling, uniformly mixing to obtain a blending spinning stock solution, and then performing spinning and crosslinking to obtain the boron nitride nanosheet modified protein-cellulose fiber.

[0055] The protein comprises at least one of soybean protein, rice protein, peanut protein, and rapeseed protein.

[0056] The amount of cellulase in the temperature conducting agent is 1-1.5% of the mass of the boron nitride nanosheet modified protein-cellulose fiber; and the response pH of the acid-sensitive liposome is less than 5.5.

[0057] The mother culture medium comprises PDA culture medium, sawdust powder, VB2, and growth promoting compound; the amount of the sawdust powder is 30% of the PDA culture medium; the amount of VB2 is 0.1% of the mass of the PDA culture medium; and the amount of the growth promoting compound is 20% of the mass of the PDA culture medium.

[0058] The growth promoting compound comprises wheat powder, ammonium sulfate, magnesium sulfate, and potassium chloride, and the mass ratio of the wheat powder, ammonium sulfate, magnesium sulfate, and potassium chloride is 5:1:0.2:0.2.

[0059] The cultivation medium comprises solid material and water, and the ratio of the solid material to water is 1:1.6.

[0060] The solid material comprises the following components in mass fraction, based on the total amount of the solid material: 98% of amino acid compound-main compound, 1% of superphosphate, and 1% of gypsum.

[0061] The main compound in the cultivation medium comprises sawdust powder, cottonseed hull, corn cob, bran, corn powder, and soybean meal, and the mass ratio of the sawdust powder, cottonseed hull, corn cob, bran, corn powder, and soybean meal is 6:5:3.6:3:1:1.

[0062] The amino acid compound in the cultivation medium comprises arginine, proline, histidine, and lysine, and the mass ratio of the arginine, proline, histidine, and lysine is 3:1:1:1.

[0063] The amino acid compound and the main compound are physically mixed in a mass ratio of 1:4 to obtain an amino acid compound-main compound.

[0064] The initial pH of the liquid fermentation culture medium is 6.0, and the inoculation amount of the liquid fermentation culture medium is 7% of the volume.

[0065] A preparation method of a yellow-green curly hair mushroom culture medium comprises the following steps:

[0066] S1, wood chip powder, VB2 and growth promoting compound are sequentially added in PDA culture medium to prepare a mother culture medium, and the mother culture medium is expanded to obtain a primary culture;

[0067] S2, after water is added in a container, wheat powder, yeast extract, potassium dihydrogen phosphate and temperature conducting agent are sequentially added and uniformly dispersed to obtain a uniform suspension, the suspension is a liquid fermentation culture medium, the primary culture in step S1 is inoculated in the liquid fermentation culture medium for further expansion to obtain a cultivation culture;

[0068] S3, the amino acid compound-main compound is used as a main material, supplemented with calcium superphosphate and gypsum, and then mixed uniformly, water is added in a material-water ratio of 1:1.6 for stirring to obtain a cultivation medium for inoculating the cultivation culture.

[0069] Based on the above content, the present application has the following specific embodiments:

[0070] Examples 1-3

[0071] The yellow-green curly hair mushroom culture medium provided in the preferred embodiments 1-3 of the present application is used for artificial domestication culture of a yellow-green curly hair mushroom strain with a preservation number of GDMCC66843 and a name of Floccularia luteovirens S4-1, and comprises a mother culture medium, a liquid fermentation culture medium and a cultivation medium.

[0072] The culture medium is prepared and used by the following method, and the specific steps are as follows:

[0073] S1, wood chip powder, VB2 and growth promoting compound are sequentially added in PDA culture medium to prepare a mother culture medium, and the mother culture medium is expanded to obtain a primary culture; wherein the addition amount of the wood chip powder is 30% of the PDA culture medium, the addition amount of VB2 is 0.1% of the mass of the PDA culture medium, and the addition amount of the growth promoting compound is 20% of the mass of the PDA culture medium, the growth promoting compound comprises wheat powder, ammonium sulfate, magnesium sulfate and potassium chloride, and the mass ratio of the wheat powder, ammonium sulfate, magnesium sulfate and potassium chloride is 5:1:0.2:0.2.

[0074] S2, after adding water in the container, add wheat flour, yeast extract, potassium dihydrogen phosphate, temperature conductor in turn, evenly disperse, get uniform suspension, the suspension is liquid fermentation culture medium, inoculate the original strain in step S1 in the liquid fermentation culture medium to further proliferate, cultivate at 25 DEG C and 10 DEG C between two fermentation target temperatures every 12 h alternately variable temperature culture 20d get cultivation species;Adjust pH, the initial pH of the liquid fermentation culture medium is 6.0, the inoculation amount of the liquid fermentation culture medium is 7% of its volume;

[0075] Among them, the temperature conductor is a pH responsive heat conducting modified fiber, including acid sensitive liposomes containing cellulase dispersion liquid inside, boron nitride nanosheet modified protein-cellulose fiber, low viscosity starch modified by low temperature anticoagulation, the acid sensitive liposomes are loaded on the boron nitride nanosheet modified protein-cellulose fiber impregnated with low viscosity starch;

[0076] The low viscosity starch modified by low temperature anticoagulation includes low viscosity tartrate starch ester and poly-N-isopropyl acrylamide grafted starch complex with a mass ratio of 3:1, there is no special requirement for the low viscosity starch during the process of impregnating the boron nitride nanosheet modified protein-cellulose fiber, the boron nitride nanosheet modified protein-cellulose fiber is completely immersed in the mixed solution of low viscosity tartrate starch ester and poly-N-isopropyl acrylamide grafted starch, and is dried after being immersed for 30 minutes;

[0077] The boron nitride nanosheet modified protein-cellulose fiber includes 50 parts of ionic liquid, 23 parts of bamboo cellulose, 8 parts of soybean protein, 0.7 parts of lactic acid oligomer and 4 parts of boron nitride nanosheet, the boron nitride nanosheet is added to the ionic liquid, heated and reacted, then the bamboo cellulose and lactic acid oligomer are added and heated and reacted, and then the soybean protein suspension is added after cooling, mixed uniformly to obtain a blended spinning dope, and the boron nitride nanosheet modified protein-cellulose fiber is obtained after spinning and activation and crosslinking of the blended spinning dope;

[0078] The amount of cellulase in the temperature conductor is 1.3% of the mass of the boron nitride nanosheet modified protein-cellulose fiber;The response pH of the acid sensitive liposome is less than 5.5, and it starts to gradually break when the pH is less than 5.5;

[0079] S3, the amino acid complex-main complex with a mass fraction of 98% is used as the main material, supplemented with 1% of superphosphate and 1% of gypsum, and then mixed uniformly, water is added and stirred uniformly according to the material-water ratio of 1:1.6 to obtain a cultivation substrate, which is used for inoculating and cultivating strains; wherein the main complex includes sawdust powder, cottonseed hulls, corn cobs, bran, corn flour and soybean meal with a mass ratio of 6:5:3.6:3:1:1; the amino acid complex includes arginine, proline, histidine and lysine with a mass ratio of 3:1:1:1; the amino acid complex and the main complex are physically mixed according to a mass ratio of 1:4 to obtain the amino acid complex-main complex. Arginine, proline, histidine and lysine all have a promoting effect on the growth of mycelium, and arginine is the best.

[0080] The component proportions of the liquid fermentation culture medium in Examples 1-3 are different, and the rest are the same, for details, see Table 1;

[0081] Table 1 Component proportions of the liquid fermentation culture medium in Examples 1-3

[0082]

[0083] Examples 4-5

[0084] Based on Example 2, the component proportions of the boron nitride nanosheet modified protein-cellulose fiber in Examples 4-5 are different from those in Example 2, and the rest are the same, for details, see Table 2;

[0085] Table 2 Component proportions of the boron nitride nanosheet modified protein-cellulose fiber in Examples 4-6

[0086]

[0087] Examples 6-7

[0088] Based on Example 2, the amount of cellulase added in the temperature conductive agent in Examples 6-7 is different from that in Example 2, and the rest are the same, for details, see Table 3;

[0089] Table 3 Amount of cellulase added in the temperature conductive agent in Examples 6-7

[0090]

[0091] Comparative Example 1

[0092] Based on Example 2, different from Example 2, the temperature conducting agent in this comparative example does not contain cellulase dispersion liquid containing acid-sensitive liposomes, and the rest is consistent.

[0093] Comparative Example 2

[0094] Based on Example 2, different from Example 2, the liquid fermentation culture medium in this comparative example does not contain a temperature conducting agent, and the rest is consistent.

[0095] Comparative Example 3

[0096] Based on Example 2, different from Example 2, the temperature conducting agent in this comparative example does not contain low-viscosity starch modified by low-temperature anticoagulation, and the rest is consistent.

[0097] Comparative Example 4

[0098] Based on Example 2, different from Example 2, the temperature conducting agent in this comparative example does not contain low-viscosity starch modified by low-temperature anticoagulation, and the rest is consistent.

[0099] Comparative Example 5

[0100] Based on Example 2, different from Example 2, the fiber in the temperature conducting agent in this comparative example does not contain boron nitride nanosheets, and the preparation method of the fiber used in this comparative example is to add bamboo cellulose and lactic acid oligomers into ionic liquid for heating reaction, then add soybean protein suspension after cooling, mix uniformly to obtain a blended spinning dope, and then perform spinning, post-activation and crosslinking to obtain protein-cellulose fiber, and the rest is consistent.

[0101] Comparative Example 6

[0102] Based on Example 2, different from Example 2, the fiber in the temperature conducting agent in this comparative example is only pure bamboo fiber, instead of boron nitride nanosheet modified protein-cellulose fiber, and no modification is performed, and the rest is consistent.

[0103] Comparative Example 7

[0104] Based on Example 2, different from Example 2, the fiber in the temperature conducting agent in this comparative example does not contain protein, and the preparation method is to add boron nitride nanosheets into ionic liquid for heating reaction, then add bamboo cellulose and lactic acid oligomers for further heating reaction, and then obtain a blended spinning dope, and then perform spinning, post-activation and crosslinking to obtain boron nitride nanosheet modified cellulose fiber, and the rest is consistent.

[0105] Comparative Example 8

[0106] Based on Example 2, different from Example 2 is that the boron nitride nanosheet modified protein-cellulose fiber in the present comparative example does not contain lactic acid oligomers; the heat transfer speed is too fast, and the temperature adaptation time is short.

[0107] Comparative Example 9

[0108] Based on Example 2, different from Example 2 is that the low-temperature anticoagulant modified low-viscosity starch in the present comparative example does not include low-viscosity starch.

[0109] Comparative Example 10

[0110] Based on Example 2, different from Example 2 is that the low-temperature anticoagulant modified low-viscosity starch in the present comparative example does not include poly-N-isopropyl acrylamide grafted starch complex.

[0111] Comparative Example 11

[0112] Based on Example 2, different from Example 2 is that the cellulase dispersion liquid in the present comparative example is not wrapped with acid-sensitive liposomes, but is directly added and used, and the rest is consistent.

[0113] Comparative Example 12

[0114] Based on Example 2, different from Example 2 is that the inoculum in step S1 is inoculated in a liquid fermentation medium for further propagation, and a cultivation species is obtained by culturing at 25°C for 20d.

[0115] Comparative Example 13

[0116] Based on Example 2, different from Example 2 is that the inoculum in step S1 is inoculated in a liquid fermentation medium for further propagation, and a cultivation species is obtained by culturing at 10°C for 20d.

[0117] Test Example 1

[0118] Test Examples 1-7 and Comparative Examples 1-13 to detect whether the cultivation species can be cultivated successfully in the corresponding cultivation medium, and the results are shown in Table 4.

[0119] The detection method is as follows: after the cultivation species is inoculated on the cultivation medium (the inoculation amount is 10% of the dry weight of the cultivation medium, and the inoculation method is the prior art), the cultivation medium is observed within 14 days to see if the strain germinates and has the phenomenon of eating the material (the germination and eating phenomenon can be seen with the naked eye, and direct observation is sufficient). If it appears, it means that the cultivation of the strain (mycelium) is successful, and at the same time, the time for the mycelium to extend into the medium (eating time) is recorded, and the results are shown in Table 4.

[0120] Table 4 Cultivation germination

[0121]

[0122] The time for the obvious eating material phenomenon visible to the naked eye in the present application can reach about 2 days at the fastest, and the liquid fermentation medium is crucial; the mycelium physical map and the corresponding micrograph of the strain after liquid fermentation in Example 2 and Comparative Examples 12 and 13 are as shown in Figures 1-3 , and the mycelium propagation under constant temperature of 25℃ or constant temperature of 10℃ is not as good as the propagation under temperature change. Example 2 is the best scheme. The white fluffy structure (cottony bloom) after germination in Example 2 is as shown in Figure 8 , which is pure in color and has no pollution.

[0123] Test Example 2

[0124] Based on Example 2, the original strain inoculated in the liquid fermentation medium is further propagated, and the change process of alternating temperature culture every 12h between 25℃ and 10℃ as two fermentation target temperatures is as shown in Figure 4 , and the mycelium growth curve is as shown in Figure 5 , which is stable and then tends to be flat, and 20d is the best culture period and obvious solid-liquid separation phenomenon appears, which is convenient for separation of mycelium.

[0125] Test Example 3

[0126] Based on Example 2, the pH of the culture medium during the further propagation of the original strain in the liquid fermentation medium is detected, and the pH gradually changes from the initial 6.0 to about 4.3.

[0127] Test Example 4

[0128] Based on Example 2, the daily growth rate (mm / d) of mycelium under different material-water ratios in the cultivation medium is detected, and the material-water ratio of 1:1.6 is the best, and the results are as shown in Figure 6 , and the corresponding cultivation physical map is as shown in Figure 7 .

[0129] Test Example 5

[0130] Based on Example 2, the effect of culture time on the extracellular enzyme activity of the liquid strain of Volvariella gloiocephala in liquid fermentation is detected, and the results are as shown in Figure 9 , wherein, Figure 9 Part A of which is a relationship diagram of amylase activity and liquid fermentation culture time; Figure 9 Part B of which is a relationship diagram of protease activity and liquid fermentation culture time; Figure 9 Part C of which is a relationship diagram of laccase activity and liquid fermentation culture time; Figure 9 Part D of which is a relationship diagram of cellulase activity and liquid fermentation culture time.

[0131] The above description is only the preferred embodiment of the present application, and is not used to limit the protection scope of the present application. Any modification, equivalent replacement and improvement made by any person skilled in the art within the spirit and principle of the present application should be included in the protection scope of the present application.

Claims

1. A culture medium for Chrysophryllum aurantiacum, characterized in that it comprises: The artificial domestication culture of the Floccularia luteovirens S4-1 strain with the preservation number GDMCC66843 and the name of yellow-green floccularia mushroom includes a mother culture medium, a liquid fermentation culture medium and a cultivation medium. The liquid fermentation culture medium includes the following components with the mass fraction: 30-40 g of wheat flour, 1-3 g of yeast extract, 0.5-1.5 g of potassium dihydrogen phosphate and 3-5 g of a temperature conducting agent per liter of solvent, and the solvent is water, and is suitable for alternating temperature culture for 20 days between the target temperatures of 25 / 10 ℃ every 12 hours. The temperature conducting agent is a pH-responsive heat-conducting modified fiber, which includes the following components: acid-sensitive liposomes containing cellulase dispersion liquid, boron nitride nanosheet modified protein-cellulose fiber and low-viscosity starch with low-temperature anticoagulation modification. The acid-sensitive liposomes are loaded on the boron nitride nanosheet modified protein-cellulose fiber impregnated with low-viscosity starch. The boron nitride nanosheet modified protein-cellulose fiber includes the following components with the weight fraction: 50 parts of ionic liquid, 20-25 parts of bamboo cellulose, 5-10 parts of protein, 0.5-1 part of lactic acid oligomer and 3-5 parts of boron nitride nanosheet, and is prepared by the following method: adding boron nitride nanosheet into ionic liquid, heating and reacting, then adding bamboo cellulose and lactic acid oligomer for further heating and reaction, subsequently adding protein suspension after cooling, uniformly mixing to obtain a blended spinning solution, and then performing spinning and crosslinking after activation to obtain the boron nitride nanosheet modified protein-cellulose fiber.

2. The culture medium for Chlorophyllum molybdites according to claim 1, characterized in that: The low-viscosity starch with low-temperature anticoagulation modification includes low-viscosity tartaric acid starch ester and poly-N-isopropyl acrylamide grafted starch complex, and the mass ratio of the low-viscosity tartaric acid starch ester to the poly-N-isopropyl acrylamide grafted starch is 3:

1.

3. The culture medium for Chlorophyllus Volvita according to claim 1, wherein: The protein includes at least one of soybean protein, rice protein, peanut protein and rapeseed protein.

4. The culture medium for Chlorophyllus Volvita according to claim 1, wherein: The amount of cellulase in the temperature conducting agent is 1-1.5% of the mass of the boron nitride nanosheet modified protein-cellulose fiber; and the response pH of the acid-sensitive liposomes is less than 5.

5.

5. The culture medium for Chlorophyllus Volvita according to claim 1, wherein: The mother culture medium includes PDA culture medium, sawdust powder, VB2 and growth promotion complex, the amount of the sawdust powder is 30% of the PDA culture medium, the amount of the VB2 is 0.1% of the mass of the PDA culture medium, and the amount of the growth promotion complex is 20% of the mass of the PDA culture medium.

6. The culture medium for Chlorophyllus Volvita according to claim 5, wherein: The growth promotion complex includes wheat flour, ammonium sulfate, magnesium sulfate and potassium chloride, and the mass ratio of the wheat flour, the ammonium sulfate, the magnesium sulfate and the potassium chloride is 5:1:0.2:0.

2.

7. The culture medium for Chlorophyllus Volvita according to claim 1, wherein: The cultivation medium includes solid material and water, and the ratio of the solid material to the water is 1:1.

6. The solid material includes the following components with the mass fraction: 98% of amino acid complex-main complex, 1% of superphosphoric acid calcium and 1% of gypsum, based on the total amount of the solid material. The main complex in the cultivation medium includes sawdust powder, cottonseed hull, corn cob, bran, corn flour and soybean meal, and the mass ratio of the sawdust powder, the cottonseed hull, the corn cob, the bran, the corn flour and the soybean meal is 6:5:3.6:3:1:

1. The amino acid compound in the cultivation substrate comprises arginine, proline, arginine, and histidine, and the mass ratio of the arginine, proline, arginine, and histidine is 3:1:1:

1. The amino acid compound and the main compound are physically mixed at a mass ratio of 1:4 to obtain the amino acid compound-main compound.

8. The culture medium for Chlorophyllus Volvita according to claim 1, wherein: The initial pH of the liquid fermentation culture medium is 6.0, and the inoculation amount of the liquid fermentation culture medium is 7% of the volume.

9. The method for preparing a culture medium of Chlorophyllus Volvita mushroom according to claim 1, characterized in that: The method comprises the following steps: S1, adding sawdust powder, VB2, and growth promoting compound in sequence in the PDA culture medium to prepare a mother culture medium, and inoculating the mother culture medium to obtain a primary culture medium; S2, adding water in a container, and then adding wheat powder, yeast extract, potassium dihydrogen phosphate, and temperature conducting agent in sequence, and uniformly dispersing to obtain a uniform suspension, wherein the suspension is a liquid fermentation culture medium, inoculating the primary culture medium in step S1 into the liquid fermentation culture medium to further inoculate to obtain a cultivation medium; S3, taking the amino acid compound-main compound as a main material, and adding calcium superphosphate and gypsum as auxiliary materials, mixing uniformly, and then adding water at a material-water ratio of 1:1.6 to stir uniformly to obtain the cultivation substrate, which is used for inoculating the cultivation medium.

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