CN115413531A - Culture material for culturing needle mushrooms and needle mushroom culturing method - Google Patents
Culture material for culturing needle mushrooms and needle mushroom culturing method Download PDFInfo
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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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- 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
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- Y02W—CLIMATE CHANGE MITIGATION TECHNOLOGIES RELATED TO WASTEWATER TREATMENT OR WASTE MANAGEMENT
- Y02W30/00—Technologies for solid waste management
- Y02W30/40—Bio-organic fraction processing; Production of fertilisers from the organic fraction of waste or refuse
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Abstract
The invention relates to the technical field of needle mushroom cultivation, and discloses a culture material for needle mushroom cultivation and a needle mushroom cultivation method, wherein the culture material comprises the following components: 35-40% of wood chips; 15-20% of bran; 25-30% of corncobs; 10-20% of bean curd skin; 10-15% of brewer's grain; selenium-carrying microsphere with selenium element content of 10-20% and 0.0055-0.04%; the water content of the culture material is 61-71%. The selenium-carrying microspheres with the slow release function are added into needle mushroom culture materials, so that selenium is slowly released into the culture materials in the whole culture period to provide nutrients containing inorganic selenium for the needle mushrooms, substance metabolism in cells converts the inorganic selenium into polysaccharide selenium and protein selenium, and the content of organic selenium in needle mushroom products is improved; and the cultivation experiment proves that: the selenium content in the needle mushroom fruiting body is in positive correlation with the selenium content in the culture material.
Description
Technical Field
The invention relates to the technical field of needle mushroom cultivation, in particular to a culture material for needle mushroom cultivation and a needle mushroom cultivation method.
Background
The flammulina velutipes is commonly called as broussonetia papyrifera, frozen mushrooms, hackberry mushrooms, winter mushrooms, golden mushrooms and anoectochilus robusta, is delicious in taste and rich in nutrition, each 100g of fresh mushrooms contains 2.72g of protein, 0.13g of fat, 5.45g of sugar, 1.77g of crude fiber, 0.83g of ash, 1.48mg of phosphorus, 0.37mg of potassium, 0.22mg of iron, 0.097mg of calcium, 0.22mg of sodium and 2.27mg of vitamin, and is rich in various amino acids necessary for human bodies, particularly high in lysine and arginine content and at the head of mushrooms, so that the flammulina velutipes is praised as 'intelligence-enhancing mushrooms' and 'intelligence-mushrooms' by people because children can promote intelligence development and healthy growth. In addition, the needle mushroom also contains substances with various physiological activities and pharmacological actions such as needle mushroom essence, polysaccharide and the like, and is a famous functional nutritional health-care food.
The trace element selenium is an essential element for human body, and is combined with some enzymes in the body to generate macromolecular compounds with biological characteristics, and the compounds can protect the body from oxidation damage and enhance immunity. Since the human body cannot synthesize selenium by itself, it needs to be taken from the outside.
The preparation of the flammulina velutipes product which has no toxic or side effect on human bodies and is rich in organic selenium improves the selenium level of food chains, and the regulation of selenium nutrition from the source is an important way for supplementing selenium elements to human bodies.
Disclosure of Invention
Catalyzing esterification of aliphatic olefine acid and glucose in an organic solvent by using immobilized lipase Novozym-435 to synthesize sugar ester with alkenyl, initiating synthesis of a glucosyl polymer by using potassium persulfate, taking the glucosyl polymer and sodium alginate as matrix materials by a complex coacervation method, wrapping sodium selenite to prepare selenium-carrying microspheres, adding the selenium-carrying microspheres into a needle mushroom culture material, absorbing nutrients in the culture material in the growth process of needle mushroom hyphae, enabling inorganic selenium to enter cells, enabling substance metabolism in the cells to combine the selenium-carrying microspheres onto macromolecular active substances, converting the selenium-carrying microspheres into polysaccharide selenium and protein selenium, and improving the content of organic selenium in needle mushroom products;
in order to achieve the purpose, the invention provides the following technical scheme:
the invention provides a culture material for culturing needle mushrooms, which comprises the following components: 35-40% of wood chips; 15-20% of bran; 25-30% of corncobs; 10-20% of bean curd skin; 10-15% of brewer's grain; selenium-carrying microsphere with selenium element content of 10-20% and 0.0055-0.04%; the water content of the culture material is 61-71%;
the preparation method of the selenium-loaded microsphere comprises the following steps:
dissolving glucose and 5-hexenoic acid in butanone, adding lipase Novozym-435 into a reaction system, adding a molecular sieve, reacting on a shaking table, filtering, evaporating the solvent to dryness, and separating and purifying by using a silica gel chromatographic column to obtain the alkenylated sugar ester;
step two, filling purified water in a sealed polymerization bottle, adding potassium persulfate and alkenyl sugar ester at the same time, introducing nitrogen, placing the mixture in a constant-temperature shaking table for reaction, filtering, and performing vacuum drying to obtain a glucose-based polymer;
dissolving sodium alginate in a beaker filled with distilled water, stirring until the sodium alginate is dissolved into a transparent colloidal solution, adding liquid paraffin and Tween 80, stirring, and standing to form a water/oil type emulsion;
dissolving the glucosyl polymer in methanol, adding sodium selenite to prepare a solution containing sodium selenite and glucosyl polymer, adjusting the pH value, dropwise adding the solution into the water/oil type emulsion under stirring, continuing stirring after the dropwise addition, adding glutaraldehyde for solidification, adding n-butyl alcohol, fully stirring, standing, centrifuging to separate out precipitates, washing with distilled water, and performing vacuum drying to obtain the selenium-loaded microspheres.
The invention provides a needle mushroom cultivation method based on a culture material for needle mushroom cultivation, which comprises the following steps:
step one, mixing raw materials according to a culture material for culturing flammulina velutipes;
step two, bottling by using a full-automatic bottling machine, wherein uniform bottling is required and the compactness is moderate; wherein, the water content of the culture material is between 61 and 71 percent;
step three, adopting high-pressure sterilization, keeping the temperature at 110-120 ℃ for 1h to kill all microorganisms and spores in the raw materials, wherein the water content of the sterilized compost is 60-70%;
after the sterilization is finished, pushing the sterilization trolley into a cooling chamber, controlling the temperature of the cooling chamber to be 13-15 ℃, and purifying air in the cooling chamber;
fifthly, inoculating by adopting an automatic inoculating machine at 20 ℃, placing the inoculating machine in a sufficiently clean inoculating room, conveying the inoculated inoculating machine to the outside of the inoculating room through a conveying belt, and transferring the inoculated inoculating machine into a culture room for culture;
a sixth step of,the temperature of the culture room is controlled to be 14-18 ℃, the humidity is controlled to be 70% -85%, and CO is added 2 The concentration is 2500-3500 mu L/L, and the culture time of the flammulina velutipes is 20-25 d;
step seven, performing mycelium stimulation treatment after the culture is finished, removing old surface strains, and performing water treatment after mycelium stimulation; wherein, the water injection amount (7-10) mL/bottle, the water content of the surface of the bottle mouth is controlled at 60-70%, and the bottle position after mycelium stimulation is moved to a bedstead of a cultivation room;
and step eight, promoting bud growth after mycelium stimulation until harvesting.
Preferably, the specific method for inducing primordial growth is as follows:
1-3 days after mycelium stimulation, maintaining the temperature of the cultivation room at 14-15 ℃, the humidity at 97-98 percent and CO 2 The concentration is 2500-3000 μ L/L, so that surface hypha can recover rapidly, but the hypha is not too dense;
maintaining the temperature of the cultivation room at 14-15 deg.C, humidity at 85-95%, and CO for 4-7 days after fungus killing 2 The concentration is 1500-2000 mu L/L, and primordium begins to form;
10 days after mycelium stimulation, the humidity of the breeding chamber is 95-97%, the material surface part is ensured to be distributed with buds, meanwhile, a plant fluorescent lamp is used for illumination, when the buds grow to be 5mm away from the bottle opening, the temperature is kept at 5-10 ℃, and CO is maintained at 2 The concentration is 4000-7000 mu L/L, the plant fluorescent lamp illuminates for 2 hours, and the illumination intensity is controlled to be 200-400 Lux;
controlling the growing room CO 17d after scratching the fungi 2 The concentration is 10000-15000 mu L/L until harvesting.
Compared with the prior art, the invention has the following beneficial technical effects:
the selenium-carrying microspheres with the slow release function are added into needle mushroom culture materials, so that selenium is slowly released into the culture materials in the whole culture period to provide nutrients containing inorganic selenium for the needle mushrooms, substance metabolism in cells converts the inorganic selenium into polysaccharide selenium and protein selenium, and the content of organic selenium in needle mushroom products is improved.
And the cultivation experiment proves that: the selenium content in needle mushroom fruiting bodies is in positive correlation with the selenium content in the culture materials, when the adding amount of the selenium-loaded microspheres (the selenium element content is 18.3%) in the culture materials is in the range of 0.02%, the biological enrichment factor of the needle mushroom fruiting bodies for selenium reaches 0.51, the organic rate reaches 86%, and the single yield of each bottle is not significantly different from the single yield of each bottle without the selenium-loaded microspheres.
Detailed Description
The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. The embodiments and features of the embodiments in the present application may be combined with each other without conflict. All other embodiments, which can be obtained by a person skilled in the art without inventive step based on the embodiments of the present invention, are within the scope of protection of the present invention.
The reagents used in the following examples are commercially available from manufacturers conventional in the art; the experimental methods used are all routine experimental methods known to those skilled in the art.
The needle mushroom test strain is platinum No. 1, and is from the scientific edible fungus research institute of Gaoyou city, jiangsu province.
Example 1
The invention provides a culture material for culturing needle mushrooms, which comprises the following components: the cultivation raw materials are wood chips and 38 percent; 18 percent of bran; 28 percent of corncob; 15% of bean curd skin; 13 percent of beer lees; selenium-carrying microsphere with selenium element content of 18.3% and 0.002%; the water content of the culture material is 65%.
The preparation steps of the selenium-loaded microspheres in example 1 are as follows:
dissolving 5.4g of glucose and 6.8g of 5-hexenoic acid in 100mL of butanone, adding 1g of lipase Novozym-435 into a reaction system, adding 38g of molecular sieve, reacting for 48 hours on a shaking table at 40 ℃ and 200r/min, filtering, evaporating the solvent to dryness, and separating and purifying by using a silica gel chromatographic column to obtain the alkenylated sugar ester;
filling 200mL of purified water in a 250mL sealed polymerization bottle, simultaneously adding 30mg of potassium persulfate and 228mg of alkenyl sugar ester, introducing nitrogen for 30min, placing the mixture in a constant-temperature shaking table at 60 ℃ and 200r/min for reaction for 12h, filtering, and drying in vacuum to obtain a glucose-based polymer;
dissolving 300mg sodium alginate in a beaker filled with 50mL distilled water, stirring until the sodium alginate is dissolved into a transparent colloidal solution, adding 50mL liquid paraffin and 20 drops of Tween 80, stirring for 30min at 25 ℃ and 200r/min, and standing to form a water/oil type emulsion;
dissolving 300mg of glucosyl polymer in 50mL of methanol, adding 400mg of sodium selenite, preparing a solution containing sodium selenite and glucosyl polymer, adjusting the pH value to 3, dropwise adding the solution into the water/oil type emulsion while stirring, continuously stirring for 30min after dripping, adding 5mL of glutaraldehyde, curing for 30min, adding 150mL of n-butyl alcohol, fully stirring, standing for 30min, centrifuging at 3000r/min to separate out precipitate, washing with distilled water, and vacuum drying at 35 ℃ to obtain the selenium-loaded microsphere.
Comparative example 1
The invention provides a culture material for culturing needle mushrooms, which comprises the following components: the cultivation raw materials are sawdust and 38 percent; 18 percent of bran; 28% of corncobs; 15% of bean curd skin; 13 percent of beer lees; the water content of the culture material is 65%.
Example 2
A needle mushroom cultivation method based on a culture material for needle mushroom cultivation comprises the following steps:
step one, mixing the raw materials according to the culture material for culturing the flammulina velutipes in the embodiment 1;
step two, bottling by using a full-automatic bottling machine, wherein uniform bottling is required and the compactness is moderate; wherein the charging amount of the plastic bottle is 1kg, and the water content of the culture material is 65%;
step three, sterilizing at high pressure, keeping the temperature at 115 ℃ for 1h to kill all microorganisms and spores in the raw materials, wherein the water content of the sterilized culture material is 64%;
after the sterilization is finished, the sterilization vehicle is pushed into a cooling chamber, the temperature of the cooling chamber is controlled to be 14 ℃, and the cooling chamber uses air to be purified;
fifthly, inoculating by adopting an automatic inoculating machine at 20 ℃, placing the inoculating machine in a sufficiently clean inoculating room, conveying the inoculated inoculating machine to the outside of the inoculating room through a conveying belt, and transferring the inoculated inoculating machine into a culture room for culture;
step six, controlling the temperature of the culture room to be 15 ℃, the humidity to be 75 percent and CO 2 The concentration is 2800 mu L/L, and the cultivation time of the flammulina velutipes is 24d;
step seven, performing mycelium stimulation treatment after the culture is finished, removing old surface strains, and performing water treatment after mycelium stimulation; wherein, the water injection amount is 8 mL/bottle, the water content of the surface of the bottle mouth is controlled at 65%, and the bottle position after mycelium stimulation is carried on a bed frame of a human cultivation room;
step eight, 2d after mycelium stimulation, keeping the temperature of the cultivation room at 15 ℃, the humidity at 98 percent and CO 2 The concentration is 2800 μ L/L to recover surface hypha rapidly, but not to make hypha too dense;
maintaining the cultivation room at 15 deg.C and 90% humidity for 5 days after fungus killing, and maintaining CO 2 At a concentration of 1800. Mu.L/L, primordia begin to form;
after mycelium stimulation for 9-12 days, the humidity of a growth room is 96%, the small buds are distributed on the surface of the material, meanwhile, a plant fluorescent lamp is used for illumination, when the small buds grow to be 5mm away from the bottle opening, the temperature is kept at 8 ℃, and CO is kept at 2 The concentration is 5000 mu L/L, the plant fluorescent lamp illuminates for 2h, and the illumination intensity is controlled at 300Lux;
controlling the growing room CO 17d after scratching the fungi 2 The concentration is 13000 mu L/L until harvesting.
Comparative example 2:
a needle mushroom cultivation method based on a culture material for needle mushroom cultivation comprises the following steps:
step one, mixing the raw materials according to the culture material for culturing the flammulina velutipes in the comparative example 1;
see example 2 for the remaining steps.
Results and analysis:
1. the single yields of enoki mushrooms in example 2 and comparative example 2 were calculated according to the formula [ single yield per bottle (g) = (total harvest quality-mycelium stimulation put in storage)/number of cultivation bottles ], and the results are shown in table 1 below.
2. Determination of selenium content
(1) And (3) determination of total selenium content: accurately weighing 0.3000g (accurate to 0.0001 g) of needle mushroom fruiting body powder into a microwave digestion tube, adding 8mL of nitric acid, placing a gasket, screwing a tank cover, placing the tube into a CEM digestion system for microwave digestion treatment (temperature 180 ℃, pressure 300MPa, power 1000W and time 30 min), taking out the digestion tube after the program is finished, removing acid, fixing the volume to 50mL, preparing a standard curve solution, and detecting the total selenium content in a sample solution by using an inductively coupled plasma mass spectrometer. And (4) calculating the biological enrichment factor of the flammulina velutipes according to a formula [ biological enrichment factor = [ selenium content of fruiting bodies/(mg/kg) ]/[ selenium content of culture materials/(mg/kg) ].
(2) And (3) determination of inorganic selenium content: accurately weighing 0.5000g (accurate to 0.0001 g) of needle mushroom powder into a 50mL centrifuge tube, adding 30mL of ultrapure water, performing ultrasonic extraction for 20min, oscillating in a water bath at 80 ℃ for 10min, cooling, centrifuging at 10000r/min for 10min, transferring supernatant to a separating funnel, repeatedly extracting with cyclohexane for 3 times, and separating out a water phase, namely an inorganic selenium extracting solution. Concentrating inorganic selenium extract to 4-7 mL, transferring into a digestion tube, adding 10mLHNO 3 And detecting the content of inorganic selenium in the sample liquid by using an inductively coupled plasma mass spectrometer.
(3) And (3) determining the content of organic selenium: calculating the organic selenium content in the needle mushroom powder according to a formula [ organic selenium content (mg/kg) = total selenium content/(mg/kg) -inorganic selenium content/(mg/kg);
the organic ratio of selenium was calculated according to the formula [ organic ratio/% = [ organic selenium content/(mg/kg) × 100]/[ total selenium content/(mg/kg) ].
The results of the above measurements are shown in tables 1 and 2 below; wherein, the table 1 shows the measurement data of the single yield and the total selenium content of the flammulina velutipes cultivated by adding and not adding the selenium-carrying microspheres in the cultivation material; table 2 shows the data of the measurement of the total selenium content, the biological enrichment factor, the inorganic selenium content, the organic selenium content and the organic selenium organization rate of the flammulina velutipes cultured by adding the selenium-loaded microspheres into the culture medium.
TABLE 1
| Example 2 (addition of selenium-loaded microspheres) | Comparative example 2 (selenium microsphere without additive) | |
| Per bottle per yield (g) | 455 | 458 |
| Total selenium content (mg/kg) | 22.95 | 0.02 |
TABLE 2
| Example 2 | |
| Total selenium content (mg/kg) | 22.95 |
| Biological enrichment factor | 0.51 |
| Inorganic selenium content (mg/kg) | 3.11 |
| Organic selenium content (mg/kg) | 19.84 |
| Organic fraction of selenium (%) | 86 |
Claims (10)
1. The culture material for culturing the needle mushrooms is characterized by comprising the following components in parts by weight: 35-40% of wood chips; 15-20% of bran; 25-30% of corncobs; 10-20% of bean curd skin; 10-15% of brewer's grain; selenium-carrying microsphere with selenium element content of 10-20% and 0.0055-0.04%; the water content of the culture material is 61-71%.
2. The culture material for needle mushroom culture according to claim 1, wherein the selenium-loaded microspheres have a selenium element content of 18.3%.
3. The culture material for needle mushroom culture according to claim 1, wherein the selenium-loaded microspheres are prepared by the following steps:
step one, dissolving glucose and 5-hexenoic acid in butanone, adding lipase into a reaction system, adding a molecular sieve, reacting on a shaking table, filtering, evaporating a solvent to dryness, and separating and purifying by using a silica gel chromatographic column to obtain an alkenylated sugar ester;
step two, filling purified water in a sealed polymerization bottle, adding potassium persulfate and alkenyl sugar ester at the same time, introducing nitrogen, placing the mixture in a constant-temperature shaking table for reaction, filtering, and performing vacuum drying to obtain a glucose-based polymer;
dissolving sodium alginate in a beaker filled with distilled water, stirring until the sodium alginate is dissolved into a transparent colloidal solution, adding liquid paraffin and tween, stirring, and standing to form a water/oil type emulsion;
dissolving the glucosyl polymer in methanol, adding sodium selenite to prepare a solution containing sodium selenite and glucosyl polymer, adjusting the pH value, dropwise adding the solution into the water/oil type emulsion under stirring, continuing stirring after the dropwise addition, adding glutaraldehyde for solidification, adding n-butyl alcohol, fully stirring, standing, centrifuging to separate out precipitates, washing with distilled water, and performing vacuum drying to obtain the selenium-loaded microspheres.
4. A culture material for flammulina velutipes-based cultivation according to claim 1, wherein the culture material comprises: 38 percent of wood dust; 18 percent of bran; 28% of corncobs; 15% of bean curd skin; 13 percent of beer lees; selenium-carrying microsphere with selenium element content of 18.3% and 0.002%; the water content of the culture material is 65%.
5. A cultivation method of needle mushrooms based on compost for cultivation of needle mushrooms as claimed in any one of claims 1 to 4, comprising the steps of:
step one, mixing raw materials according to a culture material for culturing flammulina velutipes;
step two, bottling by using a full-automatic bottling machine, wherein uniform bottling is required and the compactness is moderate; wherein, the water content of the culture material is between 61 and 71 percent;
step three, adopting high-pressure sterilization, keeping the temperature at 110-120 ℃ for 1h to kill all microorganisms and spores in the raw materials, wherein the water content of the sterilized compost is 60-70%;
after the sterilization is finished, pushing the sterilization trolley into a cooling chamber, controlling the temperature of the cooling chamber to be 13-15 ℃, and purifying air in the cooling chamber;
fifthly, inoculating by adopting an automatic inoculating machine at 20 ℃, placing the inoculating machine in a sufficiently clean inoculating room, conveying the inoculated inoculating machine to the outside of the inoculating room through a conveying belt, and transferring the inoculated inoculating machine into a culture room for culture;
step six, controlling the temperature of the culture room to be 14-18 ℃, the humidity to be 70% -85%, and CO 2 The concentration is 2500-3500 mu L/L, and the culture time of the flammulina velutipes is 20-25 d;
step seven, performing mycelium stimulation treatment after the culture is finished, removing old surface strains, and performing water treatment after mycelium stimulation; wherein, the water injection amount (7-10) mL/bottle, the water content of the surface of the bottle mouth is controlled at 60-70%, and the bottle position after mycelium stimulation is moved to a bedstead of a cultivation room;
and step eight, inducing budding and breeding after mycelium stimulation until harvesting.
6. The needle mushroom cultivation method according to claim 5, wherein the bud growth promotion method comprises: 1-3 days after fungus scratching, maintaining the temperature of the cultivation room at 14-15 ℃, the humidity at 97-98 percent and CO 2 The concentration is 2500-3000 μ L/L.
7. The needle mushroom cultivation method according to claim 5, wherein the bud growth promoting method comprises: maintaining the temperature of the cultivation room at 14-15 deg.C, humidity at 85-95%, and CO 4-7 days after mycelium stimulation 2 The concentration is 1500-2000 μ L/L.
8. The needle mushroom cultivation method according to claim 5, wherein the bud growth promotion method comprises: 9-12 days after fungus scratching, the humidity of the growth chamber is 95% -97%, the plant fluorescent lamp is used for illumination, when the buds grow to 5mm away from the bottle mouth, the temperature is kept at 5-10 ℃, and CO is kept at 2 The concentration is 4000-7000 mu L/L.
9. The needle mushroom cultivation method according to claim 8, wherein the lighting is performed by using a plant fluorescent lamp, the lighting is performed for 2 hours by using the plant fluorescent lamp, and the lighting intensity is controlled to be 200 to 400Lux.
10. The needle mushroom cultivation method according to claim 5, wherein the bud growth promoting method comprises: controlling the growing room CO 17d after scratching the fungi 2 The concentration is 10000-15000 mu L/L until harvesting.
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