CN115413530A - Liquid strain culture medium for velvet antler mushroom and method for cultivating velvet antler mushroom by using culture medium - Google Patents
Liquid strain culture medium for velvet antler mushroom and method for cultivating velvet antler mushroom by using culture medium Download PDFInfo
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Images
Classifications
-
- 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
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G18/00—Cultivation of mushrooms
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G18/00—Cultivation of mushrooms
- A01G18/40—Cultivation of spawn
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- Life Sciences & Earth Sciences (AREA)
- Mycology (AREA)
- Environmental Sciences (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Mushroom Cultivation (AREA)
Abstract
The invention discloses a liquid strain culture medium of velvet antler mushroom and a method for cultivating velvet antler mushroom by adopting the culture medium, wherein the liquid strain culture medium of velvet antler mushroom comprises the following components in parts by weight: 40-60 parts of whole wheat flour, 18-26 parts of peanut cake powder and K 2 HPO 4 1.5-2.5 parts of MgSO 4 ·7H 2 1.5-2.5 parts of O. The method for cultivating the velvet antler mushroom by adopting the velvet antler mushroom liquid strain culture medium comprises the following steps: preparing a first-level seed solution of the velvet antler mushroom, then carrying out fermentation culture on the first-level seed solution by adopting the velvet antler mushroom liquid strain culture medium to obtain a fermentation liquid, and inoculating the fermentation liquid into a cultivation material for fruiting culture. The whole wheat flour in the liquid strain culture medium of the velvet antler mushroom can provide a carbon source for the velvet antler mushroom, the peanut cake powder provides a nitrogen source for the velvet antler mushroom, and K 2 HPO 4 And MgSO 4 ·7H 2 O provides inorganic salt for the velvet antler mushroom, the cost of the culture medium is low, and the mycelium biomass of the velvet antler mushroom can be effectively improved.
Description
Technical Field
The invention relates to the technical field of edible fungi, in particular to a liquid strain culture medium for velvet antler mushroom and a method for cultivating velvet antler mushroom by adopting the culture medium.
Background
Velvet antler mushroom, known as Lyophyllum decastes (Fr.) Singer, is a rare high-grade fungus used as both medicine and food. The previous research result shows that the velvet antler mushroom fruiting body polysaccharide mainly comprises beta-1, 3-D-glucan and beta-1, 6-D-glucan, and the beta-1, 3-D-glucan can achieve the effect of inhibiting the activity of tumors by enhancing the defense function of a host; the sporophore polysaccharide and the polyphenol have good free radical scavenging capacity; the water extract of fruiting body has effects in resisting hereditary type 2 diabetes, reducing serum total cholesterol, and resisting radiation and allergy; 6-hydroxy-L-tryptophan separated from fruiting body has tyrosinase activity inhibiting effect. The velvet antler mushroom fruit body has good drug effect, contains a large amount of edible dietary fiber, has crisp and cool mouthfeel, has light taste, is popular with common people, and becomes the pleurotus eryngii. The huge demand of the domestic market for the velvet mushroom promotes the rapid development of the velvet mushroom industry, so that the velvet mushroom industry becomes a novel edible mushroom industrial cultivated variety after flammulina velutipes, pleurotus eryngii, hypsizigus marmoreus and agaricus bisporus, and production bases are mainly gathered in edible mushroom enterprises related to Shanghai, jiangsu, zhejiang and Fujian. However, the prior literature does not have a liquid fermentation medium for the velvet antler mushroom for factory production, and the commonly used medium is a general-purpose medium for liquid culture of fungi, so that the research on a liquid culture medium suitable for factory cultivation of the velvet antler mushroom is very meaningful.
Disclosure of Invention
In order to overcome the defects in the prior art, the invention aims to provide a liquid strain culture medium for the velvet antler mushroom and a method for cultivating the velvet antler mushroom by adopting the culture medium.
The technical scheme for solving the technical problems is as follows: the liquid strain culture medium for the velvet antler mushroom is provided and comprises the following components in parts by weight: 40-60 parts of whole wheat flour, 18-26 parts of peanut cake powder and K 2 HPO 4 1.5-2.5 parts and MgSO 4 ·7H 2 And 1.5-2.5 parts of O.
On the basis of the technical scheme, the invention can be further improved as follows:
further, the velvet antler mushroom liquid strain culture medium comprises the following components in parts by weight: 45-55 parts of whole wheat flour, 20-24 parts of peanut cake powder and K 2 HPO 4 1.75-2.25 parts and MgSO 4 ·7H 2 O1.75-2.25 parts.
Further, the velvet antler mushroom liquid strain culture medium comprises the following components in parts by weight: 45-52 parts of whole wheat flour, 21.2-23.5 parts of peanut cake powder and K 2 HPO 4 1.9-2.2 parts and MgSO 4 ·7H 2 O1.8-2.1 parts.
Further, the velvet antler mushroom liquid strain culture medium comprises the following components in parts by weight: 47 parts of whole wheat flour, 22 parts of peanut cake flour and K 2 HPO 4 2 parts and MgSO 4 ·7H 2 And O2 portion.
Further, the medium components further comprise: 5-10 parts of tenebrio molitor manure extract and 5-10 parts of astragalus extract; preferably 8 parts of tenebrio molitor dung extract and 6 parts of astragalus extract.
Further, the tenebrio molitor dung extract is prepared by the following method: drying the yellow mealworm feces to obtain dry yellow mealworm feces, adding water with the weight 3-6 times that of the dry yellow mealworm feces into the dry yellow mealworm feces, boiling for 15-25min, and filtering to obtain filtrate which is the yellow mealworm feces extract.
Further, the astragalus extract is prepared by the following method: pulverizing radix astragali to obtain radix astragali powder, adding 3-6 times of water into radix astragali powder, soaking for 30-50min, boiling for 20-40min, cooling, and centrifuging to obtain first supernatant and first precipitate; adding water 3-6 times the weight of the first precipitate into the first precipitate, boiling for 30-50min, cooling, centrifuging to obtain second supernatant, mixing the first supernatant and the second supernatant, and concentrating to 1/5 volume of the mixed solution.
The method for cultivating the velvet antler mushroom by adopting the velvet antler mushroom liquid strain culture medium comprises the following steps:
preparing a first-grade seed solution of the velvet antler mushroom, then carrying out fermentation culture on the first-grade seed solution by adopting the velvet antler mushroom liquid strain culture medium to obtain a fermentation liquid, and inoculating the fermentation liquid into a cultivation material for fruiting culture.
In the first-stage seed liquid preparation, the velvet antler mushroom can be cultured by adopting a conventional culture medium, such as a PDA enriched culture medium or a common edible mushroom culture medium, or velvet antler mushroom strains are firstly inoculated in a PDA enriched solid culture medium flat plate for activated culture and then added in a common edible mushroom culture medium or a PDA culture medium for liquid culture, and the like, as long as velvet antler mushroom liquid strains with higher activity can be obtained.
Further, the process of performing fermentation culture on the first-level seed liquid by adopting the velvet antler mushroom liquid strain culture medium specifically comprises the following steps: inoculating the first-class seed liquid into a liquid strain culture medium of the velvet antler mushroom according to the inoculation proportion of 1 per thousand, and controlling the aeration linear velocity Ws to be 2.0-2.2m/h, the temperature to be 23-25 ℃ and the fermentation time to be 5-8d.
Further, inoculating the fermentation liquor into the cultivation material according to the inoculation ratio of 25-30mL per bag, and harvesting after the bag filling period, the fungus bag post-maturation period and the fruiting period after hypha germination and growth. Wherein each bag of cultivation material weighs 1400g.
The formula of the cultivation material is as follows: 46% of China fir sawdust, 1% of corncobs, 12% of corn flour, 3.5% of bean stalks, 24% of bran, 5.5% of bean skins, 5% of bean pulp, 0.3% of quicklime, 2.7% of light calcium carbonate and 63-65% of water content. The raw materials in the formula are in percentage by mass.
The formula of the cultivation material can also be added with a tenebrio molitor excrement extract and an astragalus extract, wherein the mass percentages of the added tenebrio molitor excrement extract and the added astragalus extract are respectively 5% and 4%.
Further, controlling the temperature at 19-23 deg.C, relative humidity at 70% or less, carbon dioxide concentration at 3000ppm or less, and dark culturing.
Further, the temperature is controlled to be 21-25 ℃ in the fungus bag after-ripening process, the relative humidity is less than or equal to 70 percent, the carbon dioxide concentration is less than or equal to 3000ppm, and the dark culture is carried out.
Further, the fruiting stage includes primordium formation stage, pre-fruiting body development stage and post-fruiting body development stage.
And (3) an original base forming stage: the temperature is 16-18 deg.C, relative humidity is 80-90%, carbon dioxide concentration is 1500-3000ppm, and illumination intensity is 500Lx, and the device is turned on for 3sec and turned off for 30min.
Pre-development stage of fruiting body: the temperature is 15-17 deg.C, relative humidity is 80-90%, carbon dioxide concentration is 3000-5000ppm, illumination intensity is 500Lx, the condition is opened for 20sec, and the condition is closed for 30min, and the culture time of said stage is 3/4 of the development time of sporophore.
And (3) fruiting body development later stage: the temperature is 14-16 deg.C, relative humidity is natural, carbon dioxide concentration is 4000-6000ppm, illumination intensity is 500Lx, opening for 90sec, closing for 30min, and the culture time of the stage accounts for 1/4 of the fruiting body development time.
The invention has the following beneficial effects:
the whole wheat flour in the liquid strain culture medium of the velvet antler mushroom can provide a carbon source for the velvet antler mushroom, the peanut cake powder provides a nitrogen source for the velvet antler mushroom, and K 2 HPO 4 And MgSO 4 ·7H 2 O provides inorganic salt for the velvet antler mushroom. In addition, the liquid culture medium is also added with a yellow mealworm excrement extract and an astragalus extract, and after the velvet antler mushroom is cultured by adopting the culture medium, whole wheat flour, peanut cake powder, K and the like in the components of the culture medium 2 HPO 4 、MgSO 4 ·7H 2 The tenebrio molitor excrement extract and the astragalus extract can be cooperated with each other, so that the hypha biomass of the velvet antler mushroom can be effectively improved, the hypha growth is promoted, when liquid strains are inoculated to the cultivation material, the tenebrio molitor excrement extract and the astragalus extract can be added to the cultivation material, the hypha growth is promoted, the mixed bacteria pollution is inhibited, the strain planting time is shortened, and the production problem that the pollution rate is high due to the long planting period of the velvet antler mushroom is indirectly solved.
Drawings
FIG. 1 shows the effect of different culture media on the biomass of the mycelia of P.velvet antler.
Detailed Description
The culture medium of the strain and the kit used in the invention is as follows:
1. test strains
The strain of the velvet antler mushroom is preserved in the ancient field bacterial research institute of agriculture and forestry university of Fujian.
2. Reagent
Yeast powder and peptone are both purchased from Oxoid; agar powder was purchased from Chembase; glucose, sucrose, fish meal, vatamin B1, K 2 HPO 4 、MgSO 4 ·7H 2 O、ZnSO 4 、CaSO 4 The inorganic salts are purchased from chemical reagents of national drug group, inc.; the corn flour, the wheat flour and the whole wheat flour are purchased from Longlong; corncobs are purchased from deep processing of lianfeng agricultural products; soybean meal, beef extract, soybean cake powder, peanut cake powder, cottonseed cake powder and corn steep liquor are all purchased from Hongrunbaoshun culture medium raw material manufacturers.
3. Culture medium formula
PDA enriched liquid medium (1L): 200g of potato, 20g of glucose, 2g of peptone, 2g of yeast powder 2 HPO 4 2.5g，MgSO 4 ·7H 2 O1 g, vitamin B1.1 g, pH Natural. PDA enriched solid medium 20g of agar was added on a liquid medium basis.
Shake flask first-order seed liquid culture medium: 20g of glucose, 2g of peptone, 2g of yeast powder 2 HPO 4 2.5g，MgSO 4 ·7H 2 O1g, pH is natural.
The basic cultivation material formula comprises the following components: 46% of China fir sawdust, 1% of corncobs, 12% of corn flour, 3.5% of bean stalks, 24% of bran, 5.5% of bean skins, 5% of bean pulp, 0.3% of quicklime, 2.7% of light calcium carbonate and 63-65% of water content. The percentage of each raw material in the formula is mass percentage.
The following examples are given for the purpose of illustration only and are not intended to limit the scope of the invention. The examples, in which specific conditions are not specified, were carried out according to conventional conditions or conditions recommended by the manufacturer. The reagents or instruments used are not indicated by the manufacturer, and are all conventional products available commercially.
Example 1:
a liquid strain culture medium for velvet antler mushrooms comprises the following components in parts by weight: 47g/L whole wheat flour, 22g/L peanut cake flour and K 2 HPO 4 2g/L、MgSO 4 ·7H 2 O 2g/L，The pH is natural.
The preparation process of the culture medium comprises the following steps: collecting whole wheat flour 47g, peanut cake powder 22g, and K 2 HPO 4 2g、MgSO 4 ·7H 2 And O2 g, uniformly mixing, adding water to a constant volume of 1L, and sterilizing to obtain the traditional Chinese medicine.
The preparation process of the culture medium in the examples and comparative examples was identical to that described above.
Example 2:
a liquid strain culture medium for velvet antler mushrooms comprises the following components in parts by weight: 40g/L of whole wheat flour, 18g/L of peanut cake powder and K 2 HPO 4 1.5g/L、MgSO 4 ·7H 2 O1.5 g/L, pH is natural.
Example 3:
a liquid strain culture medium for velvet antler mushrooms comprises the following components in parts by weight: 60g/L of whole wheat flour, 26g/L of peanut cake powder and K 2 HPO 4 2.5g/L、MgSO 4 ·7H 2 O2.5 g/L, pH is natural.
Example 4:
a liquid strain culture medium for velvet antler mushrooms comprises the following components in parts by weight: 45g/L of whole wheat flour, 20g/L of peanut cake powder and K 2 HPO 4 1.75g/L、MgSO 4 ·7H 2 O1.75 g/L, pH is natural.
Example 5:
a liquid strain culture medium for velvet antler mushrooms comprises the following components in parts by weight: 55g/L of whole wheat flour, 24g/L of peanut cake powder and K 2 HPO 4 2.25g/L、MgSO 4 ·7H 2 O2.25 g/L, pH is natural.
Example 6:
a liquid strain culture medium for velvet antler mushrooms comprises the following components in parts by weight: 45g/L of whole wheat flour, 21.2g/L of peanut cake flour and K 2 HPO 4 1.9g/L、MgSO 4 ·7H 2 O1.8 g/L, pH is natural.
Example 7:
a liquid strain culture medium for velvet antler mushrooms comprises the following components in parts by weight: 52g/L of whole wheat flour, 23.5g/L of peanut cake flour and K 2 HPO 4 2.2g/L、MgSO 4 ·7H 2 O2.1 g/L, pH is natural.
Example 8:
a liquid strain culture medium for velvet antler mushrooms comprises the following components in parts by weight: 47g/L whole wheat flour, 22g/L peanut cake flour and K 2 HPO 4 2g/L、MgSO 4 ·7H 2 O2 g/L, yellow mealworm excrement extract 5g/L and astragalus extract 5g/L.
The yellow mealworm excrement extract is prepared by the following method: drying the yellow mealworm feces to obtain dry yellow mealworm feces, adding water with the weight 5 times that of the dry yellow mealworm feces into the dry yellow mealworm feces, boiling for 20min, and filtering to obtain filtrate which is the yellow mealworm feces extract.
The astragalus extract is prepared by the following method: pulverizing radix astragali to obtain radix astragali powder, adding water 5 times the weight of radix astragali powder into radix astragali powder, soaking for 40min, boiling for 30min, cooling, and centrifuging to obtain first supernatant and first precipitate; adding 5 times of water into the first precipitate, boiling for 40min, cooling, centrifuging to obtain second supernatant, mixing the first supernatant and the second supernatant, and concentrating to 1/5 volume of the mixed solution.
Example 9:
a liquid strain culture medium for velvet antler mushrooms comprises the following components in parts by weight: 47g/L whole wheat flour, 22g/L peanut cake flour and K 2 HPO 4 2g/L、MgSO 4 ·7H 2 O2 g/L, 10g/L of tenebrio molitor dung extract and 10g/L of astragalus extract.
The yellow mealworm excrement extract is prepared by the following method: drying the yellow mealworm feces to obtain dry yellow mealworm feces, adding water with the weight 5 times that of the dry yellow mealworm feces into the dry yellow mealworm feces, boiling for 20min, and filtering to obtain filtrate which is the yellow mealworm feces extract.
The astragalus extract is prepared by the following method: pulverizing radix astragali to obtain radix astragali powder, adding water 5 times the weight of radix astragali powder into radix astragali powder, soaking for 40min, boiling for 30min, cooling, and centrifuging to obtain first supernatant and first precipitate; adding 5 times of water into the first precipitate, boiling for 40min, cooling, centrifuging to obtain second supernatant, mixing the first supernatant and the second supernatant, and concentrating to 1/5 volume of the mixed solution.
Example 10:
a liquid strain culture medium for velvet antler mushrooms comprises the following components in parts by weight: 47g/L whole wheat flour, 22g/L peanut cake flour and K 2 HPO 4 2g/L、MgSO 4 ·7H 2 O2 g/L, yellow mealworm excrement extract 8g/L and astragalus extract 6g/L.
The yellow mealworm excrement extract is prepared by the following method: drying the yellow mealworm feces to obtain dry yellow mealworm feces, adding water with the weight 5 times that of the dry yellow mealworm feces into the dry yellow mealworm feces, boiling for 20min, and filtering to obtain filtrate which is the yellow mealworm feces extract.
The astragalus extract is prepared by the following method: pulverizing radix astragali to obtain radix astragali powder, adding water 5 times the weight of radix astragali powder into radix astragali powder, soaking for 40min, boiling for 30min, cooling, and centrifuging to obtain first supernatant and first precipitate; adding 5 times of water into the first precipitate, boiling for 40min, cooling, centrifuging to obtain second supernatant, mixing the first supernatant and the second supernatant, and concentrating to 1/5 volume of the mixed solution.
Comparative example 1:
a liquid strain culture medium for velvet antler mushroom comprises the following components in parts by weight: glucose 20g/L, peptone 2g/L, K 2 HPO 4 2.5g/L、MgSO 4 ·7H 2 O1 g/L, pH is natural.
Comparative examples 2 to 8:
comparative examples 2 to 8 differ from comparative example 1 in that: the glucose was replaced with whole wheat flour, and the amounts of whole wheat flour in comparative examples 2-8 were 10g/L, 20g/L, 30g/L, 40g/L, 50g/L, 60g/L, and 70g/L, respectively.
Comparative examples 9 to 15:
comparative examples 9 to 15 differ from comparative example 1 in that: peptone was substituted for peanut meal, and the amounts of peanut meal in comparative examples 9-15 were 2g/L, 6g/L, 10g/L, 14g/L, 18g/L, 22g/L, 26g/L, respectively.
Comparative examples 16 to 20:
comparative examples 16 to 20 and comparative examplesThe difference is that: k 2 HPO 4 In different amounts, K in comparative examples 16 to 20 2 HPO 4 The content of (b) is respectively 0.5g/L, 1g/L, 1.5g/L, 2g/L and 3g/L.
Comparative examples 21 to 25:
comparative examples 21 to 25 differ from comparative example 1 in that: mgSO (MgSO) 4 ·7H 2 Different contents of O, mgSO in comparative examples 21 to 25 4 ·7H 2 The O content is 0.5g/L, 1.5g/L, 2g/L, 2.5g/L and 3g/L respectively.
Experimental example 1 Effect of different media on the biomass of the mycelia of P.velvet
1. Preparation of first-order seed liquid in shake flask
Inoculating the strain of the velvet antler mushroom to a PDA enriched solid culture medium flat plate, and culturing at 25 ℃ for about 20 days until hyphae grow over the flat plate. Punching 30 holes on a flat plate by using a puncher (with the diameter of 1.0 mm), selecting a bacterium block into a shaking bottle first-stage seed liquid culture medium (containing a magnetic stirrer) by using an inoculator, culturing for 4d at 25 ℃,150rpm, taking out the shaking bottle from the 4 th d, placing the shaking bottle on the magnetic stirrer at 800rpm for 30min, scattering bacterium balls, and continuously culturing for 4d to obtain a first-stage seed liquid.
2. The primary seed solution was inoculated into the culture media of examples 1-7 and comparative examples 1-25 at 10% inoculum size, cultured in shake flask at 25 ℃ at 150rpm for 8d, filtered through 100 mesh gauze to collect the pellet, washed 3 times with RO water, dried to constant weight at 60 ℃ and 3 replicates per treatment set-up to determine hyphal biomass.
As is clear from the results of the measurements, the hyphal biomass in examples 1 to 7 varied little, and was at the maximum of 33.91g/L, in example 1, all of which were 30 to 33.91g/L. While comparative examples 1 to 25 are considerably different from examples 1 to 7, and comparative examples 1 to 25 are based on comparative example 1, and the one-factor design was conducted, as can be seen from FIG. 1, when whole wheat flour was used as a carbon source for the medium and the amount of addition was 40 to 60g/L, the biomass of dried mycelia reached a maximum (13.10 g/L) (FIG. 1A). When peanut cake powder was used as the nitrogen source of the medium, the biomass of dried mycelia reached the maximum value (24.65 g/L) when the amount of the additive was 18-26g/L (FIG. 1B). With K 2 HPO 4 With MgSO 4 ·7H 2 When O is an inorganic salt of the culture medium, K 2 HPO 4 With MgSO 4 ·7H 2 The O addition was between 1.5 and 2.5g/L and the dry mycelial biomass reached its maximum value (FIGS. 1C and 1D).
3. Whole wheat flour, peanut cake flour, K 2 HPO 4 And MgSO 4 ·7H 2 The influence of O on the biomass of the hypha of the velvet antler mushroom is different, and the factors influencing the biomass of the hypha of the velvet antler mushroom are as follows: whole wheat flour, peanut cake flour, K 2 HPO 4 And MgSO 4 ·7H 2 O, the interaction order is: whole wheat flour and K 2 HPO 4 Whole wheat flour and MgSO 4 ·7H 2 O, peanut cake flour and K 2 HPO 4 、K 2 HPO 4 And MgSO 4 ·7H 2 O, whole wheat flour and peanut meal (peanut meal and MgSO) 4 ·7H 2 O). When the whole wheat flour is 45-52g/L and the peanut cake flour is 21.2-23.5g/L, the whole wheat flour is 45-51.5g/L and K 2 HPO 4 1.9-2.2g/L, whole wheat flour 45.2-51.5g/L and MgSO 4 ·7H 2 The biomass of the dry hypha has a maximum value within the range of O1.8-2.1 g/L; the whole wheat flour has obvious interaction with the peanut cake powder and the magnesium sulfate heptahydrate. There is little interaction between the other factors, mainly by the single factor itself determining the dry mycelial biomass.
4. The invention also researches the influence of other different carbon sources, nitrogen sources and inorganic salts on the biomass of the hypha of the velvet antler mushroom, wherein the carbon sources are corncobs, corn flour, wheat flour and cane sugar; the nitrogen source is soybean meal, soybean cake powder, beef extract, fish meal, corn steep liquor, peptone and cottonseed cake powder; the inorganic salt is CoCl or CuSO 4 、ZnSO 4 、CaSO 4 、MnSO 4 、FeSO 4 NaCl and KCl were used in place of the carbon source, nitrogen source and inorganic salt in example 1. After the carbon source, the nitrogen source and the inorganic salt in example 1 are respectively replaced by the carbon source, the nitrogen source and the inorganic salt, the biomass of the hyphae of the velvet antler mushroom is reduced, the biomass of the hyphae is sequentially reduced after the whole wheat flour as the carbon source in example 1 is respectively replaced by cane sugar, wheat flour, corn flour and corncob, and the biomass of the hyphae in example 1 is 1.8 times of the biomass of the hyphae after the cane sugar is replaced by cane sugarIn example 1, the hyphal biomass was 2.2 times that of the hyphae obtained by replacing wheat flour. In the embodiment 1, when the nitrogen source peanut meal cake is replaced by cottonseed cake powder, corn steep liquor, fish meal, beef extract, soybean cake powder and soybean meal powder, the hypha biomass is reduced in sequence, and after the nitrogen source peanut meal cake is replaced by the cottonseed cake powder, the hypha biomass is reduced by 0.2 times, and the corn steep liquor is reduced by 0.3 times. When the inorganic salt K in example 1 2 HPO 4 And MgSO 2 4 ·7H 2 Replacing O with KCl, naCl and FeSO 4 、MnSO 4 、CaSO 4 、ZnSO 4 、CuSO 4 After CoCl, hyphal biomass is reduced in sequence, and after KCl is replaced, hyphal biomass is reduced by 0.4 times and is replaced by CuSO 4 CoCl inhibits hyphal growth, especially after CoCl replacement, which is slower.
And adding the yellow mealworm excrement extract and the astragalus extract into the culture medium, as in the examples 8-10, the yellow mealworm excrement extract and the astragalus extract are added on the basis of the example 1, when the yellow mealworm excrement extract and the astragalus extract are added, the hypha growth can be promoted, and the hypha biomass can be increased, the hypha biomass can be measured according to the culture process of the examples 8-10, and the hypha biomass is respectively 39.21g/L, 40.2g/L and 42.81g/L after the culture of the examples 8-10.
Test example 2 production application
The 1000L fermentation tank is used for carrying out amplification fermentation on the velvet antler mushroom, and the specific process is as follows: 0.8L of the first-order seed liquid was prepared according to the method of test example 1, and inoculated into 800L of the liquid medium, and the fermentation was carried out for 7d at a ventilation linear velocity Ws of 2.0m/h and a temperature of 24 ℃, at which time the pellet was free-settled for 3h and the pellet settlement coefficient was about 95%. Microscopic examination results show that: the average diameter of the cenospheres cultured in the example 1 is 2.63 +/-0.14 mm, the density of the cenospheres reaches 1070 +/-70/mL, the average diameter of the cenospheres cultured in the example 10 is 3.16 +/-0.12 mm, and the density of the cenospheres reaches 1890 +/-60/mL. However, when the velvet antler mushroom is fermented and cultured in the embodiment 10, the fermentation time is 5 days, and the vitality of the fungus balls is the strongest, so that when the culture medium in the embodiment 10 is adopted in a factory cultivation experiment, the fermentation time is 5 days.
The experimental process can be divided into the following groups according to different culture media and cultivation materials:
a group of: the culture medium of example 1 is adopted for liquid fermentation, and the culture medium is used as a basic culture medium formula.
Two groups are as follows: the culture medium of example 10 was used in liquid fermentation, and the cultivation material was the basic cultivation material formulation.
Three groups: the culture medium of example 10 was used for liquid fermentation, and 5% of tenebrio molitor dung extract was added to the formula of the basic culture medium.
Four groups: the culture medium of example 10 was used for liquid fermentation, and 4% of astragalus extract was added to the formula of the basic culture medium.
Five groups are as follows: the culture medium of example 10 was used for liquid fermentation, and 5% of tenebrio molitor manure extract and 4% of astragalus extract were added to the formula of the basic culture medium.
Inoculating the fermented strain into a cultivation bag according to an inoculation ratio of 25-30 mL/bag (each bag is 1400g of cultivation material), and harvesting after hyphae germinate, hyphae full bag, fungus bag and mature period and fruiting period; wherein, the fruiting period comprises: primordia forming stage, pre-sporocarp development stage and post-sporocarp development stage.
The culture parameters at each stage are as follows:
controlling the temperature to be 22 ℃, the relative humidity to be less than or equal to 70 percent and the carbon dioxide concentration to be less than or equal to 3000ppm in the hypha growth process, and carrying out dark culture;
controlling the temperature to be 24 ℃, the relative humidity to be less than or equal to 70 percent and the carbon dioxide concentration to be less than or equal to 3000ppm in the fungus bag after-ripening process, and carrying out dark culture;
and (3) an original base forming stage: the temperature is 16 ℃, the relative humidity is 85%, the carbon dioxide concentration is 2000ppm, and the illumination intensity is 500Lx, the opening is carried out for 3sec, and the closing is carried out for 30min;
pre-development stage of fruiting body: the temperature is 16 ℃, the relative humidity is 85%, the carbon dioxide concentration is 4000ppm, the illumination intensity is 500Lx, the opening is carried out for 20sec, the closing is carried out for 30min, and the culture time of the stage accounts for 3/4 of the development time of the sporocarp;
and (3) fruiting body development later stage: the temperature is 14 ℃, the relative humidity is natural, the carbon dioxide concentration is 5000ppm, the illumination intensity is 500Lx, the opening time is 90sec, the closing time is 30min, and the culture time of the stage accounts for 1/4 of the development time of the sporocarp.
The culture times of the various groups of strains are compared in the following table:
from the above, when the liquid culture medium provided by the invention is used for culturing the velvet antler mushroom, the hypha biomass of the velvet antler mushroom can be effectively improved, the hypha growth is promoted, and the cultivation time of the velvet antler mushroom is shortened.
The above description is only for the purpose of illustrating the preferred embodiments of the present invention and is not to be construed as limiting the invention, and any modifications, equivalents, improvements and the like that fall within the spirit and principle of the present invention are intended to be included therein.
Claims (10)
1. The liquid strain culture medium for the velvet antler mushrooms is characterized by comprising the following components in parts by weight: 40-60 parts of whole wheat flour, 18-26 parts of peanut cake powder and K 2 HPO 4 1.5-2.5 parts and MgSO 4 ·7H 2 And 1.5-2.5 parts of O.
2. The velvet antler mushroom liquid strain culture medium according to claim 1, characterized by comprising the following components in parts by weight: 47 parts of whole wheat flour, 22 parts of peanut cake flour and K 2 HPO 4 2 parts and MgSO 4 ·7H 2 And O2 portion.
3. The velvet antler mushroom liquid strain culture medium according to claim 1 or 2, further comprising: 5-10 parts of tenebrio molitor dung extract and 5-10 parts of astragalus extract.
4. The velvet antler mushroom liquid strain culture medium according to claim 3, further comprising: 8 parts of tenebrio molitor dung extract and 6 parts of astragalus extract.
5. The velvet antler mushroom liquid strain culture medium according to claim 3, wherein the tenebrio molitor manure extract is prepared by a method comprising: drying the yellow mealworm feces to obtain dry yellow mealworm feces, adding water with the weight 3-6 times that of the dry yellow mealworm feces into the dry yellow mealworm feces, boiling for 15-25min, and filtering to obtain filtrate which is the yellow mealworm feces extract.
6. The velvet antler mushroom liquid strain culture medium according to claim 3, wherein the astragalus extract is prepared by the following method: pulverizing radix astragali to obtain radix astragali powder, adding 3-6 times of water into radix astragali powder, soaking for 30-50min, boiling for 20-40min, cooling, and centrifuging to obtain first supernatant and first precipitate; adding water 3-6 times the weight of the first precipitate into the first precipitate, boiling for 30-50min, cooling, centrifuging to obtain second supernatant, mixing the first supernatant and the second supernatant, and concentrating to 1/5 volume of the mixed solution.
7. The method for cultivating velvet antler mushroom by using the velvet antler mushroom liquid strain culture medium as claimed in any one of claims 1 to 6, characterized by comprising the steps of:
preparing a velvet antler mushroom primary seed solution, then carrying out fermentation culture on the primary seed solution by using the velvet antler mushroom liquid strain culture medium of any one of claims 1 to 6 to obtain a fermentation liquid, and inoculating the fermentation liquid into a cultivation material for fruiting culture.
8. The cultivation method according to claim 7, wherein the fermentation broth is inoculated into the cultivation material at an inoculation ratio of 25-30mL per bag, and is harvested after hypha germination and growth to a bag filling stage, a fungus bag post-ripening stage and a fruiting stage.
9. The cultivation method according to claim 8, wherein the temperature is controlled to be 19-23 ℃, the relative humidity is less than or equal to 70%, the carbon dioxide concentration is less than or equal to 3000ppm during the hypha growth process, and dark cultivation is performed;
controlling the temperature to be 21-25 ℃ in the fungus bag after-ripening process, controlling the relative humidity to be less than or equal to 70 percent and controlling the carbon dioxide concentration to be less than or equal to 3000ppm, and carrying out dark culture.
10. The cultivation method according to claim 8, wherein the fruiting stage includes an primordial formation stage, a pre-fruiting body development stage and a post-fruiting body development stage;
and (3) an original base forming stage: the temperature is 16-18 ℃, the relative humidity is 80-90%, the carbon dioxide concentration is 1500-3000ppm, the illumination intensity is 500Lx, the opening is carried out for 3sec, and the closing is carried out for 30min;
pre-development stage of fruiting body: the temperature is 15-17 deg.C, relative humidity is 80-90%, carbon dioxide concentration is 3000-5000ppm, illumination intensity is 500Lx, opening is for 20sec, closing is for 30min, and the culture time of the stage accounts for 3/4 of the fruiting body development time;
and (3) fruiting body development later stage: the temperature is 14-16 ℃, the relative humidity is natural, the carbon dioxide concentration is 4000-6000ppm, the illumination intensity is 500Lx, the opening time is 90sec, the closing time is 30min, and the culture time of the stage accounts for 1/4 of the development time of the sporocarp.
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| CN116171797A (en) * | 2023-04-04 | 2023-05-30 | 福建恒绿生物科技有限公司 | Cultivation method of velvet mushrooms |
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