CN109122051B - Culture method of boletus aereus solid strain - Google Patents
Culture method of boletus aereus solid strain Download PDFInfo
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- CN109122051B CN109122051B CN201811011350.2A CN201811011350A CN109122051B CN 109122051 B CN109122051 B CN 109122051B CN 201811011350 A CN201811011350 A CN 201811011350A CN 109122051 B CN109122051 B CN 109122051B
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G18/00—Cultivation of mushrooms
- A01G18/40—Cultivation of spawn
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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
Abstract
The invention relates to a culture method of boletus aereus solid strains. The culture method of the boletus aereus solid strain comprises the following steps: culturing a solid strain of boletus nigricans in the following culture medium, wherein the culture medium comprises the following components: 6-18 parts of liquidambar formosana tree fermentation product, 6-18 parts of giant junge grass, 6-18 parts of bagasse, 20-40 parts of barley, 20-40 parts of sorghum, 20-40.0 parts of red soil, 1.0-3.0 parts of gypsum powder, and MgSO (MgSO) as raw material40.1 to 0.15 part by weight of KH2PO40.1 to 0.15 portion. According to the invention, the sweetgum tree fermentation product is added into the culture medium to generate a large amount of effective fertilizer and humus, so that the looseness and air permeability of the culture medium can be increased, and holes do not need to be punched during the culture of bolete, thereby solving the problem of invisible pollution caused by punching.
Description
Technical Field
The invention relates to the technical field of strain culture, in particular to a culture method of a boletus aereus solid strain.
Background
Bolete nigricans (Phlebopus portentosus), also known as Phlebopus portentosus, is a precious tropical facultative mycorrhizal edible fungus, and is distributed in Srilanca, Vietnam, Indonesia, Thailand, Brazil, Mexico, Australia, New Zealand, Yunnan, Guangxi and Hainan of China. The boletus aereus has fresh and tender quality, delicious taste, rich nutritive value, high protein and low fat, is rich in 17 kinds of essential amino acid for human body and rich mineral elements such as phosphorus, potassium, calcium, magnesium, iron, zinc and the like which are necessary for human body, is good health food, has good medicinal value and is deeply loved by consumers. However, the natural picking quantity of boletus nigricans is very small, and the market demand cannot be met. The boletus nigricans are the only boletus species which are recognized at present and have few evolutions in the boletus edible fungi, can leave host trees for nutrition and saprophytic life, are cultivated in mushroom houses by a saprophytic fungus cultivation method, and have high yield.
The solid strain is the only production cultivar for industrially and massively cultivating the boletus nigricans at present. Since the liquid spawn cultured in the triangular flask is not suitable for large-area cultivation, the liquid spawn cultured in the fermentation tank can not be applied to production at present. Because the substrate is compact and the hypha grows slowly, the solid strain bottle needs to be perforated in the middle. However, because the anti-pollution capability of bolete is poor, even in the later stage of cultivation, as long as mould spores enter the holes of the strain bottles or the strain bags, the mould spores can be fixedly planted on the culture materials, and before inoculation, the moulds in the holes can not or very difficultly be inspected and removed, so that the mould spores become invisible pollution of cultivation, and huge pollution hidden trouble is brought to production; on the other hand, the contaminated culture medium prolongs the period of fungus cultivation and increases the cultivation cost. Therefore, the culture medium which is free from invisible mould pollution and suitable for industrial culture of boletus aereus is urgently needed in production.
In view of the above, the present invention is particularly proposed.
Disclosure of Invention
The invention aims to provide a culture method of boletus aereus solid strains, a culture medium used by the culture method is loose and porous, the contents of fertilizer and humus are high, and holes do not need to be punched when boletus aereus is cultured, so that the problem of invisible pollution caused by punching is solved, the pollution rate is low, only less than 1 per thousand, the culture period of hyphae is short, and the matrix can grow over in 30 days at most.
In order to achieve the above purpose, the invention provides the following technical scheme:
the culture method of the boletus aereus solid strain comprises the following steps of culturing the boletus aereus solid strain in the following culture medium, wherein the culture medium comprises the following components: based on the weight portion, the weight portion of the material,
compared with the prior art, the key point of the invention is that the sweetgum tree fermentation product is adopted, so that the organic matters in the fermentation product are decomposed into a large amount of effective fertilizer and humus, and the ratio of the effective fertilizer to the humus is optimized, thereby increasing the loose air permeability of the culture medium, and even the culture of boletus aereus does not need to be perforated, thereby solving the problem of invisible pollution caused by perforation.
Under the same culture condition, the pollution rate of the culture method of the invention is only less than 1 per mill when the bolete is cultured, and the pollution rate of the conventional fermentation product without the sweetgum tree is as high as about 5 percent.
In addition, the growth period of hyphae is greatly shortened because the culture medium is loose and breathable, and the hyphae overgrow the substrate after 25-30 days of culture and are used for expanding the inoculation culture bottle. Under the same conditions, the growth cycle of the conventional culture medium is more than 40 days.
In addition, the culture medium used for culturing the liquidambar formosana wood can contain trace elements, inorganic salts, vitamins, soil and other auxiliary materials and other nutritional ingredients necessary for bolete nigricans besides the liquidambar formosana tree fermentation medium.
Jujuncao is one of high-yield and high-quality fungi, and is used as culture material for culturing 49 edible fungi and medicinal fungi, such as shiitake mushroom, ganoderma lucidum, etc.
The bagasse is the residue left after sugar squeezing, contains 2% of sugar, and the fiber is crushed to 1.0 cm-2.0 cm in length. The bagasse of the present invention is preferably fermented bagasse.
The pennisetum hydridum and the bagasse can improve the physical structure of the culture medium and increase the nutrition required by the growth of hypha. The invention not only utilizes the conventional action of the Jujun grass and the bagasse, but also utilizes the synergistic action of the Jujun grass and the sweetgum tree fermentation product to improve the growth speed of the strains.
The preparation method of the culture medium can be further improved, and the details are as follows.
Preferably, the sweetgum tree fermentation product is prepared by the following method:
enabling the sweetgum trees to be subjected to primary pile fermentation for 2.5-4 months, and keeping the temperature of the center of a pile at 55-75 ℃ during the primary pile fermentation; and then crushing, and carrying out secondary pile fermentation for 0.5-1.5 months, wherein the temperature of the center of the pile is kept at 45-55 ℃ during the secondary pile fermentation.
Preferably, the sweetgum trees are wood chips with the thickness of 0.2 cm-0.5 cm, the width of 0.8 cm-1.0 cm and the length of 0.8 cm-1.0 cm.
The liquidambar formosana tree is crushed before fermentation, so that the fermentation efficiency can be improved.
Preferably, the comminution after the first pile fermentation is: crushing into particles with the thickness of 0.2 cm-0.5 cm, the width of 0.4 cm-0.6 cm and the length of 0.4 cm-0.6 cm.
On one hand, the fermentation efficiency is improved by the crushing, on the other hand, the propagation efficiency of the aerobic bacteria is improved by crushing and turning, and further, the variety and the content of the fertilizer in the culture medium are optimized.
In the invention, the temperature of the first pile fermentation can be any value of 55-75 ℃, such as 55 ℃, 60 ℃, 65 ℃, 70 ℃ and 75 ℃, and can be constant temperature or controlled in a small range. Any time of 2.5-4 months, such as 3 months, 3.5 months and the like, of the first pile fermentation.
In the invention, the temperature of the second pile fermentation can be any value in the range of 45-55 ℃, such as 45 ℃, 46 ℃, 47 ℃, 48 ℃, 49 ℃, 50 ℃, 51 ℃, 52 ℃, 53 ℃ and 54 ℃, and can be constant temperature or controlled in a small range. Any time of 0.5-1.5 months of the first pile fermentation, such as 10 days, 20 days, half month, 40 days and the like.
Preferably, the mass ratio of the pennisetum hydridum to the bagasse to the sweetgum tree fermentation product is 1-2: 1-3, preferably 1: 1: 1.
preferably, the water content of the culture medium is 51-55 wt%.
Too high water content is easy to breed and pollute, and too low is not beneficial to the growth of hypha.
Preferably, the pH of the culture medium is 4.0-6.0.
Preferably, the medium comprises the following components: based on the weight portion, the weight portion of the material,
preferably, the medium comprises the following components: based on the weight portion, the weight portion of the material,
the culture medium is mixed, bottled, sterilized and cooled, and then inoculated into liquid spawn balls cultured in advance, and the humidity is less than or equal to 50 percent, the temperature is 27-30 ℃, and the temperature is CO2Culturing for 30 days under the condition of the concentration of 1500 ppm-2500 ppm, and obtaining the high-quality solid strain.
The strain particles cultured by the method are uniform, the size of the strain particles is 0.2-0.5 cm multiplied by 0.4-0.5 cm, after the strain particles are inoculated into the culture bottle, the strain particles can uniformly fall into the holes in the middle of the culture bottle, so that the upper part, the middle part and the lower part of the culture bottle are provided with strains, the strains on the upper part, the middle part and the lower part of the culture bottle germinate and eat materials simultaneously in the initial culture period, and the hyphae grows over the substrate in the culture bottle for 25-30 days and enters the earth covering growth period.
More preferably, when the solid strain is cultured, the number of the mold spores in the air environment is controlled, and the air cleanliness is detected by a bacteria sedimentation detection method, so that the number of the mold spores is less than or equal to 1 per culture dish.
In summary, compared with the prior art, the invention achieves the following technical effects:
(1) the liquidambar formosana trees subjected to special fermentation treatment are added into the culture medium of the boletus nigra, so that the loose and porous performance of the culture medium is enhanced, and holes do not need to be punched when the culture medium is used, so that the pollution rate during culture is reduced;
(2) the synergistic effect of the sweetgum trees, the pennisetum sinese roxb and the bagasse improves the growth speed of the black beef liver hyphae;
(3) the proportion of the substrate, other nutrient components and auxiliary materials in the culture medium is optimized, and the growth speed of boletus aereus hyphae is further improved;
(4) the cultured strains have uniform particles, and can accelerate the covering speed of the hyphae, reduce the pollution rate and shorten the culture period.
Detailed Description
The technical solutions of the present invention will be described clearly and completely with reference to the following detailed description, but those skilled in the art will understand that the following described examples are some, not all, of the examples of the present invention, and are only used for illustrating the present invention, and should not be construed as limiting the scope of the present invention. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention. The examples, in which specific conditions are not specified, were conducted under 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
The first step, solid seed culture medium formula:
the fermentation product of the sweetgum trees is prepared by the following steps:
pulverizing the sweetgum into wood chips with the thickness of 0.2-0.5 cm, the width of 0.8-1.0 cm and the length of 0.8-1.0 cm, stacking the wood chips into a stock pile with the height of 2 meters, the width of 3 meters and the length of 10 meters, keeping the central temperature of the stock pile at 65-75 ℃, and stacking and fermenting for 3 months; the wood grains fermented for 3 months are crushed into grains with the thickness of 0.2 cm-0.5 cm, the width of 0.4 cm-0.6 cm and the length of 0.4 cm-0.6 cm and the size similar to that of the wheat grains, and then secondary fermentation is carried out, the central temperature of a stock pile is kept at 45-55 ℃, and the stock pile is fermented for 1 month, so that the wood grains are fully softened and decomposed.
The Jujun grass is dried in the sun, crushed into short fibers with the width of 0.3 cm-0.5 cm and the length of 0.4 cm-0.8 cm, and drenched with water 3-5 days before use.
Crushing bagasse fiber into 1.0-2.0 cm long material with water content of 45-50%, and fermenting regularly for 1-3 months.
The preparation method comprises the following steps:
accurately weighing barley and jowar, soaking for 10 hr, and draining; the fermented product of the sweetgum tree, the pennisetum sinese roxb and bagasse fiber are accurately measured according to the volume; accurately weighing other components, adding all the raw materials into a stirring barrel, adding a proper amount of water, uniformly stirring, and measuring the water content of the matrix to be 51% -55% to obtain the solid seed culture matrix.
And step two, bottling (bagging) the culture material, sterilizing and inoculating.
1100mL of plastic bottle or plastic corner-folded bag is used as a culture container, the culture material is filled into the plastic bottle or plastic corner-folded bag, and after bottling or bagging, the middle part of the bottle or bag is not perforated. Then sterilizing at 121 ℃ for 100 minutes, cooling, inoculating liquid strains, and inoculating the strains in an amount of 40mL per bottle or bag.
Culturing the solid strain mycelium: setting the humidity of culture room to be less than or equal to 50%, the temperature to be 27 ℃, and CO2Culturing hypha at 1500ppm, strictly controlling the number of mould spores in the culture room to be less than or equal to 1 per culture dish, culturing for 25-30 days, and allowing the hypha to grow over the substrate to obtain the solid strain.
Inoculating the cultured solid strain into culture bottles, wherein the inoculation amount of each bottle is 50 g. The strain particles are uniform, the size of the strain particles is 0.2-0.5 cm multiplied by 0.4-0.5 cm, the strain particles uniformly fall into the hole in the middle of the cultivation bottle, strains are arranged on the upper part, the middle part and the lower part of the cultivation bottle, the strains on the upper part, the middle part and the lower part of the cultivation bottle germinate and eat materials at the same time in the initial cultivation period, the hypha grows over the substrate after 25 days of cultivation, and the soil covering growth period is started.
Compared with the non-uniform strain particles or the large strain particles, the culture bottle needs to be cultured for 40 days, and the hyphae can grow over the solid strains, the culture period of the culture bottle is shortened by 10 days, the utilization rate of a culture room is improved, and the production cost is reduced.
The pollution rate of the culture bottle in the whole fungus culture period is 1 per mill, and compared with the pollution rate of 5 percent of the solid seed inoculation culture bottle cultured by the conventional matrix, the pollution is greatly controlled.
Examples 2 to 4
The difference from the example 1 lies in the formulation of the solid seed culture medium, as shown in table 1, other culture conditions and fermentation conditions of the sweetgum trees are the same as those of the example 1, after the obtained solid seed culture is inoculated into a culture bottle, hypha of the culture bottle grows over the substrate after being cultured for 27 days, and the pollution rate is less than 0.8 per thousand.
TABLE 1 culture Medium formulation for each example
Example 5
The difference from the example 1 is only that the fermentation conditions of the sweetgum trees are different, and the specific difference is as follows:
pulverizing the sweetgum into wood chips with the thickness of 0.2-0.5 cm, the width of 0.8-1.0 cm and the length of 0.8-1.0 cm, stacking the wood chips into a stock pile with the height of 2 meters, the width of 3 meters and the length of 10 meters, keeping the central temperature of the stock pile at 55-65 ℃, and stacking and fermenting for 4 months; the wood grains fermented for 4 months are crushed into grains with the thickness of 0.2 cm-0.5 cm, the width of 0.4 cm-0.6 cm and the length of 0.4 cm-0.6 cm and the size similar to that of the wheat grains, and then secondary fermentation is carried out, the central temperature of a stock pile is kept between 50 ℃ and 55 ℃, and the stock pile is fermented for 15 days, so that the wood grains are fully softened and decomposed.
After the solid strains are inoculated into the culture bottle, hypha grows over the substrate in the culture bottle in 28 days, and the pollution rate is 0.8 per mill.
Example 6
The culture substrate was bottled (bagged), sterilized, inoculated, and hyphae cultured as in example 1, except that: a 1300mL plastic bottle for the culture container, a humidity of less than or equal to 50%, a temperature of 29 ℃, and CO2The method comprises the steps of culturing hypha growth at the concentration of 2500ppm, strictly controlling the number of mould spores in a culture room to be less than or equal to 1 per culture dish, culturing for 26 days, enabling the hypha to grow over a substrate to obtain a solid strain, inoculating a culture bottle, and enabling the pollution rate to be 0.9 per thousand.
Example 7
The culture medium was bottled (bagged), sterilized, inoculated, and hypha cultured in the same manner as in example 1, except that the culture container was a 1500mL plastic bottle.
The cultured strain is inoculated into a culture bottle, hypha grows over the substrate within 30 days, and the pollution rate is below 0.8 per mill.
Example 8
The culture substrate was bottled (bagged), sterilized, inoculated, and hyphae cultured as in example 1, except that: the culture vessel was a 1650mL plastic bottle, and the formulation of the solid seed medium was as follows:
inoculating the cultured solid strain into a culture bottle, wherein the substrate is full of mycelia in 26 days, and the pollution rate of the culture bottle is below 0.8 per mill.
Comparative example 1
The difference from example 1 is only that the sugar cane bagasse is not contained, and the solid medium formula is as follows under the same other conditions:
the hypha grows over the substrate within 30 days during the culture, and the pollution rate is 1 per mill.
Comparative example 2
Because the matrix is loose and has poor air permeability, the hypha of the solid strain grows slowly, and the hypha grows over the matrix after 35 days of culture. Solid seed inoculation cultivation bottle, because the bacterial granule is big inhomogeneous, large granule bacterial can not fall into the bottom of cultivation and influence the hypha to germinate and eat the material, plus the material face front cover is not tight, and the pollution rate 8 thousandths, increased 8 times, it can overgrow matrix to cultivate 40 days hypha, gets into the earthing and goes out the mushroom, and the fungus culture period has increased 10 days.
Finally, it should be noted that: the above embodiments are only used to illustrate the technical solution of the present invention, and not to limit the same; while the invention has been described in detail and with reference to the foregoing embodiments, it will be understood by those skilled in the art that: the technical solutions described in the foregoing embodiments may still be modified, or some or all of the technical features may be equivalently replaced; and the modifications or the substitutions do not make the essence of the corresponding technical solutions depart from the scope of the technical solutions of the embodiments of the present invention.
Claims (4)
1. The culture method of the boletus aereus solid strain is characterized in that the boletus aereus solid strain is cultured in the following culture medium, and the culture medium comprises the following components: based on the weight portion, the weight portion of the material,
the sweetgum tree fermentation product is prepared by the following method:
enabling the sweetgum trees to be subjected to primary pile fermentation for 2.5-4 months, and keeping the temperature of the center of a pile at 55-75 ℃ during the primary pile fermentation; then crushing, and carrying out secondary pile fermentation for 0.5-1.5 months, wherein the temperature of the center of the pile is kept at 45-55 ℃ during the secondary pile fermentation;
in the culture medium, the mass ratio of the pennisetum hydridum to the bagasse to the sweetgum tree fermentation product is 1-2: 1-3;
the Jujun grass is prepared by the following method: drying in the sun, crushing into short fibers with the width of 0.3-0.5 cm and the length of 0.4-0.8 cm, and spraying water for prewetting 3-5 days before use;
the bagasse is prepared by the following method: crushing bagasse fiber into 1.0-2.0 cm long material with water content of 45-50%, and fermenting regularly for 1-3 months;
the culture medium is mixed, bottled, sterilized and cooled, and then inoculated into liquid spawn balls cultured in advance, and the liquid spawn balls are cultured at the temperature of 27-30 ℃ and the humidity of less than or equal to 50 percent and CO2Culturing for 25-30 days under the condition of the concentration of 1500-2500 ppm, and obtaining the high-quality solid strain.
2. The method for culturing solid strains of boletus nigricans according to claim 1, wherein the sweetgum trees are wood chips with a thickness of 0.2cm to 0.5cm, a width of 0.8cm to 1.0cm and a length of 0.8cm to 1.0 cm.
3. The method for culturing solid strains of boletus nigricans according to claim 1, wherein the pulverization is: crushing into particles with the thickness of 0.2 cm-0.5 cm, the width of 0.4 cm-0.6 cm and the length of 0.4 cm-0.6 cm.
4. The method for culturing solid boletus edulis strain according to claim 1, wherein the amount of mold spores in the air environment is controlled during culturing of the solid boletus edulis strain, and the air cleanliness is detected by a sedimentation bacteria detection method, so that the number of the mold spores is less than or equal to 1/culture dish.
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| CN108207487A (en) * | 2018-01-26 | 2018-06-29 | 云南强丰农业科技有限公司 | A kind of cultural method of agaric |
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| JP2007045757A (en) * | 2005-08-10 | 2007-02-22 | Takinou Filter Kk | Planting material |
| CN103508772A (en) * | 2012-06-21 | 2014-01-15 | 新疆永华生物科技开发有限公司 | Culture material for industrial cultivation of Pleurotus eryngii |
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Denomination of invention: A solid culture method of Boletus nigricans Effective date of registration: 20220714 Granted publication date: 20210615 Pledgee: China Agricultural Development Bank Xishuangbanna Dai Autonomous Prefecture Branch Pledgor: JINGHONG HONGZHEN AGRICULTURAL SCIENCE AND TECHNOLOGY Co.,Ltd. Registration number: Y2022530000019 |
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Date of cancellation: 20230711 Granted publication date: 20210615 Pledgee: China Agricultural Development Bank Xishuangbanna Dai Autonomous Prefecture Branch Pledgor: JINGHONG HONGZHEN AGRICULTURAL SCIENCE AND TECHNOLOGY Co.,Ltd. Registration number: Y2022530000019 |
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Denomination of invention: A Culture Method for Solid Strain of Black Bull Liver Fungus Effective date of registration: 20230811 Granted publication date: 20210615 Pledgee: China Agricultural Development Bank Xishuangbanna Dai Autonomous Prefecture Branch Pledgor: JINGHONG HONGZHEN AGRICULTURAL SCIENCE AND TECHNOLOGY Co.,Ltd. Registration number: Y2023530000055 |