CN111034538A - Method for culturing black fungus by utilizing mulberry branches - Google Patents

Abstract

The invention discloses a method for culturing black fungus by utilizing mulberry branches, which comprises the steps of firstly selecting a culture environment, then crushing and preparing mulberry branches, determining a culture season, selecting black fungus varieties, preparing a culture base material, culturing, managing, harvesting and finally clearing the field after harvesting. According to the method, the black fungus is cultivated by utilizing the mulberry branches, on one hand, the waste mulberry branches generated by silkworm breeding are utilized, so that the method is environment-friendly and economical, and the overall economic value of the mulberry is increased; on the other hand, the content of DNJ, calcium and selenium elements of the black fungus cultivated by using the mulberry branches as the cultivation raw material is higher than that of the black fungus cultivated by using common miscellaneous trees, and the mulberry and the fungus are organically combined, so that a low-carbon economic new mode with maximized benefit and no pollution is formed.

Description

Method for culturing black fungus by utilizing mulberry branches

Technical Field

The invention belongs to the technical field of agriculture, and particularly relates to a method for culturing black fungus by utilizing mulberry branches.

Background

The Yuexi county is 'the village of silkworm mulberry in China', the silkworm mulberry comprehensive index is the first of the whole provinces living in 9 years continuously, the existing mulberry field reaches 7 ten thousand mu, a large amount of waste mulberry branches are generated every year, the beautifying effect on the environment of the Yuexi countryside is great, and how to change the waste mulberry branches into valuable becomes a great problem in the industrial development of the Yuexi countryside. Yuexi local farmers try to mechanically crush ramulus mori and then cultivate black fungus in artificial bags, and the selenium content of each kilogram of ramulus mori black fungus is detected to be 0.17mg, so that the requirement of selenium-enriched food is met, the trace elements of calcium and iron are far superior to those of common black fungus, and the ramulus mori black fungus is rich in selenium, thick, easy to soak, tender, strong in glutinousness and good in taste and has the quality which is rapidly accepted by consumers.

Because the mulberry contains special components such as polyphenols, flavonoids, alkaloids and the like, the cultivated black fungus has the functions of reducing blood sugar and fat, inhibiting alcohol, preventing cancer, increasing immunity and the like, the mulberry branch black fungus has thick and large meat quality and soft and glutinous mouthfeel compared with the common black fungus, and the mulberry branch black fungus contains various amino acids and various trace elements such as calcium, iron, zinc, selenium and the like which are beneficial to a human body through detection, so the mulberry branch black fungus has high nutritive value and has a health-care function. The regular consumption of the health-care food is beneficial to reducing blood viscosity and preventing arteriosclerosis, myocardial infarction and cerebral thrombosis. Is a daily food material for restaurants, hotels and households, and has large market demand.

Disclosure of Invention

The invention aims to provide a method for culturing black fungus by utilizing mulberry branches, which is characterized in that the black fungus is cultured by utilizing the mulberry branches, the waste mulberry branches can be about 4500kg per mu, 450kg of dry black fungus can be produced, and the yield per mu can reach 36000 yuan according to the current market price of 80 yuan/kg. The black fungus is planted on the mulberry branches discarded in silkworm breeding, so that on the basis of meeting the planting requirement of the black fungus, the utilization rate of the mulberry branches is effectively improved, the current situation that forest trees in a black fungus planting area are seriously consumed is effectively relieved, the silkworm economic industry chain is extended, the rural environment is protected, and the economic benefit of the silkworm industry is improved.

The specific technical scheme of the invention is as follows:

a method for culturing black fungus by utilizing mulberry branches is characterized by comprising the following steps:

step one, selection of a cultivation environment: the cultivation field is selected to have good ecological condition, flat terrain, clean surrounding environment, no pollution source and good water quality, and reaches the national sanitary standard;

step two, mulberry branch crushing and material preparation: selecting dry and mildew-free mulberry twigs with bark, pulverizing into 0.2-0.8cm mulberry twig powder with a pulverizer, or directly pulverizing fresh mulberry twigs, and drying in the sun for use to prevent mildew during storage;

step two, cultivation season: the inoculation time can be generally 3-4 months;

step three, variety selection: selecting a variety which is suitable for local weather characteristics, high in yield, high in quality, strong in stress resistance and strong in market competitiveness;

step four, the formula of the culture medium: 87-89 parts of mulberry twig chips, 9-11 parts of bran, 0.8-1.2 parts of gypsum, 0.8-1.2 parts of lime and 50-55 parts of water;

step five, cultivation

5.1 preparation of the fungus stick

5.2. And (3) sterilization: performing normal-pressure steam sterilization, stacking the material rods in a shape like a Chinese character 'jing' or a bottle pile, reserving gaps of 1-2 cm between the material rods, reserving gaps for every 3-5 layers to facilitate steam circulation, keeping the central material temperature to be more than 98 ℃ within 4-5 hours after beating cold air, and keeping for 12-14 hours to ensure the sterilization effect;

5.3, taking out of the pot and cooling: cleaning, killing insects and sterilizing the cooling field in advance; after sterilization, naturally cooling to below 50-60 ℃ and discharging from a stove, wherein the raw material bag is easy to burst when the temperature is too high; lightly taking and lightly placing the material rods out of the stove, and moving the material rods into a sterilized cooling field to be cooled to natural temperature;

5.4. inoculation: before inoculation, the inoculation box is disinfected by aerosol disinfectant, and the surfaces of both hands and instruments are disinfected by 75% alcohol; cooling the material bag to below 28 ℃, selecting a blocking strain to be inoculated and slightly higher than the surface of the hole, and then sleeving a bag and tying the bag;

5.5. mycelium culture: suspension type fungus bag cultivation

The growth of the fungus hyphae is greatly influenced by the culture temperature and humidity, the fungus hyphae need to be distinguished and mastered according to different growth periods for normal growth of the fungus hyphae, and the fungus bags are inoculated and then moved into a culture field for suspension type stacking; the whole culture process is roughly divided into three stages: germination period, growth period, and maturation period; checking regularly, and turning the pile every 10-15 days; under normal conditions, the fungus sticks grow over white hyphae from 45 days to 60 days;

5.6. ear emergence management;

step six, harvesting

When the ear is eight-nine percent mature, the ear stem becomes thin, the ear lobe is spread and slightly droops, a small amount of spores are generated on the compound surface, the picking can be carried out when dew is not dry and the ear is in a damp and soft state in sunny days, impurities are removed, and the ear is dried in the sun;

seventhly, clearing the field after the mining

Cleaning impurities after harvesting, stopping spraying water for 5-7 days to promote hypha recovery, starting spraying fine water on 8 days to keep ear sticks moist, and spraying water again to grow out of ears after new ear buds are formed.

Further, in the third step, the black fungus is selected from one of black fungus 916, black wei 5 or shennong 7.

Further, step 5.1 specifically comprises: pre-wetting the main material, uniformly mixing with bran, gypsum and lime, adding water while stirring, fully stirring, bagging within 5 hours after stirring, respectively packaging into bags with 15cm × 55cm polyethylene bags, fastening the bag openings, inspecting the walls of the bacteria bags, and ensuring no thinning, abrasion or puncture.

Further, in step 5.5, germination stage: controlling the temperature to be 26-28 ℃ before 10 days after inoculation, controlling the room temperature to be about 22-24 ℃ after 10-20 days, and frequently checking and paying attention to the stack temperature, increasing the stack temperature and increasing the ventilation;

and (3) growing period: after 20 days of inoculation, the temperature is preferably controlled to be about 22 ℃, when the white mycelium grows to the diameter of 10cm-15cm around the inoculation hole, an outer layer of a bag is removed to increase oxygen and ventilate, the growth of the mycelium is promoted, meanwhile, pile turning is carried out again after 10-15 days, and the fungus sticks infected with the mixed fungi are eliminated;

and (3) mature period: after 35-40 days after inoculation, the mycelium gradually enters a physiological maturation stage, the room temperature is preferably controlled at 20-22 ℃, ventilation adjustment is mainly performed by opening and closing doors and windows, and bacteria burning caused by overhigh temperature is avoided; when white protruding buds appear after mycelia grow over the bag, cutting 2cm openings at two ends of the stick to promote the development of sporophore; at this time, the sesame can enter the sesame shed.

Further, step 5.6 specifically comprises: (1) selecting an earfield: the method has the advantages that the method is good in wind gating, sufficient in sunlight, convenient in water source, free of pollution source, capable of preventing waterlogging and used as an earfield;

(2) and (3) ear field construction: in a field ear field, drainage ditches with the depth of 30cm are arranged at the periphery of the field ear field, a ridge bed is shaped like a turtle back, the width is 140-150 cm, the length is not limited, and supports with 3 rows in the longitudinal direction, the height is 30-35 cm and the line spacing is 40-50 cm are erected by small wood (bamboo) rods or iron wires; after disinfecting and killing insects, the ridge-up bed is paved with hay or straws and a sunshade net, and a micro-spray pipe is installed;

(3) puncturing and ear-stimulating: puncturing holes on a sunny day with the temperature below 25 ℃, puncturing 150-180 ear outlet holes with the depth of 1-1.5 cm around fungus sticks by using a special puncturing machine for sterilized black fungus, stacking the fungus sticks after puncturing according to a Chinese character jing shape or a triangle shape, reducing the stacking amount in unit area, and simultaneously strengthening ventilation management; controlling the temperature below 25 ℃ and the humidity at about 80% during the fungus culturing period after puncturing, covering a plastic film for heat preservation when the humidity is lower than 20 ℃, ventilating every day, and accelerating the puncturing to culture the fungus for 7-10 days, recovering the mycelium of the puncturing, and discharging the mycelium out of the field;

(4) arranging the fields: generally, in the middle 10 th month to the middle 11 th month, clear or cloudy weather is selected for the arrangement; filling water to moisten the ear field one day before the drainage; when the mushroom sticks are arranged on a field, the mushroom sticks lean against the support at an angle of 60-70 degrees with the ground and are uniformly distributed, and the distance between the mushroom sticks and the support is 10-15 cm; 7000 to 8000 bars are arranged per mu in a field ear field;

(5) and (3) ear field management: the principle of 'alternation of wetting and drying' is adopted, less spraying is carried out in rainy days, and more spraying is carried out in sunny days and dry days; or the lug is in a leather shape and is sprayed more, and the lug is in a blister shape and is sprayed less or not sprayed; after the fungus sticks are arranged, the fungus sticks are kept from losing water, a small amount of fine water is sprayed for many times, and the relative humidity is kept between 80% and 90%; after young ears are formed and grow into small ears, water is sprayed for 1-2 times every day, and the water spraying amount is increased along with the growth of the ears so as to promote the growth of the young ears; the water spraying amount in the earning period is correspondingly increased along with the growth of the fruiting body, and the dry-wet difference is enlarged, namely dry days and wet days, the dry-wet difference alternately promotes the growth, and the relative humidity is kept between 90% and 95%; if continuous rainy days occur, covering measures are needed; if the water is accumulated in the bar inner connecting sheet and the bag, the bag is taken out and the ear is produced.

Has the advantages that:

the method utilizes mulberry branches to culture the black fungus, about 4500kg of waste mulberry branches can be utilized per mu, 450kg of dry black fungus can be produced, 80 yuan/kg is achieved according to the current market price, and the yield per mu can reach 36000 yuan. The black fungus is planted on the mulberry branches discarded in silkworm breeding, so that on the basis of meeting the planting requirement of the black fungus, the utilization rate of the mulberry branches is effectively improved, the current situation that forest trees in a black fungus planting area are seriously consumed is effectively relieved, the silkworm economic industry chain is extended, the rural environment is protected, and the economic benefit of the silkworm industry is improved. On one hand, the residual waste mulberry twigs after silkworm breeding are utilized, so that the environment is protected, the energy is saved, and the overall economic value of the mulberry is increased; on the other hand, the content of DNJ, calcium and selenium elements of the black fungus cultivated by using the mulberry branches as the cultivation raw material is higher than that of the black fungus cultivated by using common miscellaneous trees, and the mulberry and the fungus are organically combined, so that a low-carbon economic new mode with maximized benefit and no pollution is formed.

In the specific implementation process of the invention, the mulberry leaves are used for silkworm breeding, the mulberry branches and the silkworm excrement are used for cultivating edible fungi, and the crude protein, crude fat and various mineral elements in the fungus chaff can be used as compost, fuel, feed, fertilizer and the like for recycling, so that the green ecological cycle of the mulberry leaf silkworm breeding, the mulberry branch seed fungus and the fungus chaff fertilizer mulberry is realized. Closely combines the silkworm breeding industry and the edible fungus industry, and realizes diversified development and maximum utilization of resources.

Detailed Description

The technical solution of the present invention will be described in further detail with reference to examples.

A method for culturing black fungus by using mulberry branches comprises the following steps:

step one, selection of a cultivation environment: the cultivation field is selected to have good ecological condition, flat terrain, good ventilation condition, clean surrounding environment, no pollution source and good water quality, and reaches the national sanitary standard;

step two, mulberry branch crushing and material preparation: selecting dry and mildew-free mulberry twigs with bark, pulverizing into 0.2-0.8cm mulberry twig powder with a pulverizer, or directly pulverizing fresh mulberry twigs, and drying in the sun for use to prevent mildew during storage;

step three, cultivation season: the inoculation time can be selected from 3-4 months and 9-10 months.

Step three, variety selection: selecting one of varieties which are suitable for local climate characteristics, high in yield, high in quality, strong in stress resistance and market competitiveness, such as black fungus 916, Heiwei No. 5 or Shennong No. 7, as a strain;

step four, the formula of the culture medium: 88% of mulberry branch scraps, 10% of bran, 1% of gypsum, 1% of lime and 60-65% of culture medium humidity;

step five, cultivation: and (5) carrying out greenhouse three-dimensional cultivation.

51. Preparing liquid strains: the liquid strain is prepared from mother strain, shake flask and large tank.

(one), mother seed

1. Formulation (liquid spawn mother culture usually adopts enrichment medium)

200g of potato, 20g of glucose, 3g of yeast extract, 2g of peptone, 1g of monopotassium phosphate, 0.5g of magnesium sulfate, 20g of agar, natural pH value and 1000ml of fixed solution.

2. Preparation of test tube Medium

2.1 preparing nutrient solution: cleaning potato, peeling, cutting into slices, adding 1200ml water, cooking with rice cooker to crisp but not rotten degree, filtering with four layers of gauze to obtain juice, and dissolving 1000 ml. Heating and adding other culture materials, and slowly adding agar while stirring.

2.2 subpackaging test tubes: and after the culture materials are fully melted, subpackaging the hot culture materials into clean test tubes by using a long funnel, wherein the loading amount of each test tube is one fourth to one fifth of the volume. After cooling, the culture medium is solidified, no condensed water is in the tube wall, a cotton plug is plugged, and every five or seven tubes are bound into a bundle by rubber bands and wrapped by newspaper.

2.3 autoclaving: sterilizing with a portable pressure cooker under high pressure, wherein cold air in the cooker is discharged and sterilized at 121-. After the sterilization is finished, the pointer is reset to zero, the pot cover is opened to leave a small gap, and the test tube is taken out after the tampon is baked by using the waste heat in the pot.

2.4 swinging inclined plane: the sterilized culture medium is placed on an inclined plane when the culture medium is hot, the length of the inclined plane is one half to two thirds of that of the test tube, and the inclined plane is covered with towel gauze after being placed so as to slowly cool the test tube, thereby preventing a large amount of condensed water from being formed in the wall of the test tube. The prepared mother culture blank medium is cultured for 3 days at the temperature of 25 ℃, and the mother culture blank medium can be used as soon as possible without mixed bacteria.

3. Inoculation by rotating tube

And (3) sterilizing the original mother seeds, the test tube culture medium and the inoculation tool, then putting the sterilized mother seeds, the test tube culture medium and the inoculation tool into an ultra-clean workbench type inoculation box, and opening a fan and an ultraviolet lamp for sterilization for 45 minutes. Cutting the original mother seeds into small pieces of 0.5 x 0.2cm in an alcohol flame protection area, and inoculating the small pieces of the original mother seeds into the center of a blank test tube culture medium. Here, it is to be noted that: taking few aerial hyphae on one hand and performing tip extraction on the other hand.

4. Culturing

And sticking a label on the inoculated mother test tube, putting the mother test tube on a plate, and culturing the mother test tube in a dark environment at 25 ℃. The test tubes with abnormal pigments and mixed bacteria colonies are eliminated in time after the examination in time at the initial stage of the culture and the growth speed is slow. Storing in 0-5 deg.C refrigerator when the hypha grows to half.

(II) Shake flask

1. Formulation of

400g of potato, 40g of glucose, 3g of monopotassium phosphate, 1.5g of magnesium sulfate,

The pH value is natural and 1600ml of solution is fixed.

2. Making of

Selecting a proper shake flask and a proper stirrer according to the type and the size of the large tank, washing the large tank by using the shake flask, making a cotton plug, and binding the newspaper for later use. Cleaning potato, peeling, weighing 400g, slicing, adding 1800ml water, decocting until it is crisp but not rotten, and filtering with four layers of gauze to obtain juice. Adding other culture materials, heating until the culture materials are fully melted, filtering out impurities, dissolving in 1600ml, and subpackaging into a shake flask with the volume of one third of the shake flask volume. Wiping the bottle mouth, plugging the cotton plug, wrapping the newspaper, and sterilizing by using an autoclave in the same way as the test tube.

3. Inoculation of

According to the germination condition of the mother seeds, taking out one mother seed from a refrigerator one day or several days in advance, putting the mother seed in an environment at 25 ℃ for revival, and putting the mother seed and a shake flask into a super clean workbench for sterilization for 45 minutes during inoculation. Removing newspaper, loosening cotton plug, and cutting hyphae at the tip of mother seed into small pieces of 0.2 × 0.1cm in alcohol flame protection zone. Selecting more than ten blocks, putting into a shake flask, plugging a cotton plug, and wrapping newspaper.

4. Culturing

Placing the inoculated shake flask at 25 ℃ for static culture for one day, and culturing on a stirrer in a stirring way when the inoculated truffle hyphae start to germinate. The rotation speed of the stirrer depends on the number and shape of the hyphae, the whole stirrer is slow firstly and then fast, the stirrer can shake the bottles for about 7 days generally, and if the stirrer is not used immediately, the stirrer is placed in a refrigerator for storage at 0-5 ℃ (the storage period is one month).

(III) Large tank

1. Formulation (Large tank capacity 600 l)

3.6kg of soybean meal, 3.6kg of corn meal, 5.4kg of glucose, 0.54kg of monopotassium phosphate, 0.27kg of magnesium sulfate and 30g of liquid antifoaming agent.

2. Making of

2.1 inspection and cleaning of large tanks: before seed production, the large tank is subjected to gas inspection. Air is pumped into the large tank, so that the pressure in the large tank is 0.1, and the pressure is unchanged or slightly reduced after ten hours according to the change of the pressure, thereby indicating that the large tank passes the gas detection. In addition, whether the filter element is loosened, whether the filter element returns or not and whether the filter element mildews or not are checked, and the filter element can be used only after being qualified. The large tank must be thoroughly cleaned after each use or before the next use to remove the pellets, clumps, feed solution and other contaminants from the tank walls.

2.2 air extinguishing of big pot: when a new large tank is used for the first time, when the previous tank is infected with mixed bacteria, when the large tank is not used for a long time, and when a new variety is replaced, the large tank is required to be emptied and killed. The large tank is filled with water to the central line of the test mirror, and the heater is started to heat. Firstly, discharging cold air in the large tank, then closing all valves, raising the temperature and the pressure, raising the temperature to 100 ℃, opening an inoculation port valve under the pressure of 0.05MPa, and discharging the cold air in the tank. The inoculation valve is closed, and the temperature reaches 121-.

2.3 preparation of Large jar of culture Medium: weighing the culture materials, dissolving soybean meal and corn flour in cold water, grinding into slurry with a pulping machine, and dissolving glucose, potassium dihydrogen phosphate and magnesium sulfate in warm water. Adding 300l of water into the big tank, pouring the culture materials into the big tank, adding the defoaming agent, supplementing water, covering the tank cover tightly, simultaneously feeding gas into the pipe, opening the inoculation port valve, and closing the breathing valve.

2.4 Sterilization of the culture medium: and (3) introducing steam, turning down the air inlet valve to slightly introduce air into the large tank to reduce the sound, and closing the inoculation port valve and the air inlet valve when the inoculation port valve exhausts the atmosphere. When the pressure of the large tank is increased to 0.1 pressure and the temperature reaches 121 ℃, the sterilization timing is started, and the heat preservation and pressure maintaining are carried out for 1 hour. Sterilizing the filter for 20 min, and maintaining for 30-40 min. Slightly opening the valve of the inoculation port 2-3 minutes before the sterilization, sterilizing the inoculation port, and then closing the steam valve. After the inoculation opening is sterilized, the inoculation opening is protected by alcohol cotton, and the air supply valve is opened to blow the filter element dry. And opening the circulating water cooling culture solution, gradually opening the breathing valve, blowing air to the large tank after the filter element is dried, and closing the circulating water when the temperature of the large tank is reduced to 25 ℃.

3. Inoculation of

Placing inoculating tools such as shake flask into a seed room, opening ozone and ultraviolet lamp for sterilization for 45 min, and inoculating after ozone reduction. The shake flask inoculation pipe and the large jar inoculation port are sterilized by 75% alcohol, and absorbent cotton soaked in 95% alcohol is wound on the inoculation port. Igniting the flame ring, placing the shake flask inoculation tube above the flame, removing newspaper and cotton plug on the copper tube with tweezers, inserting the copper tube into the inoculation port, opening the valve of the inoculation port, and pouring the original seeds into a large tank. The stock in the flask was poured into a large jar before the flame extinguished. The copper pipe is rapidly drawn out, and the inoculation port valve is closed. Removing the iron wire ring and the alcohol cotton, quickly plugging the 75% alcohol cotton into the inoculation port, pricking the membrane for protection, and completing the inoculation.

4. Culturing

Opening a breathing valve to stabilize the pressure in the tank at 0.01-0.03MPa, and allowing the culture liquid to have a light fragrance along with the extension of the culture period after culturing for 48-72h, wherein the fragrance of the culture liquid discharged by a respirator is gradually reduced, and the culture liquid is replaced by a light bacteria liquid fragrance in the later culture period, and the concentration of the mycelium balls and the viscosity of the bacteria liquid are gradually increased. The liquid strain of black fungus is generally inoculated for 5-6 days, and the strain activity is strongest at the 6 th day. Here, it is to be noted that: the large tank must be sampled before inoculation, and the sampling culture is generally carried out 24 hours before inoculation.

52. Preparing a mushroom stick:

⑴ the large and hard impurities such as thick branches, stones, cloth ends and iron blocks in the mulberry branch scraps are screened out by a vibrating screen, and are prevented from being brought into a stirrer or a bagging machine.

⑵ adding the required ingredients (bran, gypsum, lime, etc.) into the culture medium during first-stage stirring according to the formula of the culture medium, and blending the culture medium according to the technical standard (humidity 60-65%, pH 7-9).

⑶ opening the first-stage stirring and feeding port, feeding the culture material into the second-stage stirring, and feeding the culture material into a bagging machine for bagging by a material distributor after the second-stage stirring of the culture material is completed.

⑷ bagging should be moderate in tightness, consistent in height, even in weight, and smooth in material surface, the bagging should be lightly handled in the bagging and carrying process to avoid the contamination of the mixed bacteria caused by puncturing the bacteria bags, the inspector should inspect the humidity and pH value of each batch of compost, the bagging standard execution and other conditions, and make inspection records, and the bagged bacteria sticks must be put into a sterilization cabinet for high-temperature sterilization on the same day to prevent the compost from being acidified.

53. And (3) sterilization: the material rods are stacked in a shape like a Chinese character 'jing' or a bottle pile by adopting normal-pressure steam for sterilization, a gap of 1cm-2cm is reserved between the material rods, a gap is reserved between every 3-5 layers of the material rods so as to facilitate the circulation of steam, the temperature of the central material is enabled to reach more than 98 within 4-5 hours after the cold air is tapped, and the central material is continuously kept for 12-14 hours so as to ensure the sterilization effect.

54. Taking out of the pot and cooling: cleaning, killing insects and sterilizing the cooling field in advance. After the sterilization is finished, the temperature is naturally reduced to below 50-60 ℃ and the material bag can be taken out of the stove, and the material bag is easy to burst when the temperature is too high. And lightly taking and placing the material rods out of the stove, and moving the material rods into a sterilized cooling place to be cooled to the natural temperature.

55. Liquid strain inoculation:

⑴ before inoculation, all the articles and equipment required by inoculation are put into the proper position of the inoculation chamber, the inoculation chamber is disinfected by ozone 3 hours in advance, and dust-falling treatment is carried out by disinfectant water, so as to ensure the concentration of micro-dust and bacterial particles in the inoculation chamber to be within the safe inoculation range.

⑵ the inoculator needs to have good personal hygiene, clean and tidy clothes, no nail, no cosmetics with strong taste, no hair growth, strong aseptic operation consciousness, and strict compliance with various regulations and requirements of inoculation.

⑶ the inoculator needs to wash hand with disinfectant, wear special aseptic clothes and shoes cover, wear aseptic mask, headgear and gloves, enter the air shower room for 1 min, remove most of dust and bacteria, and enter the inoculation room for formal inoculation.

⑷ the door is closed immediately after entering the inoculation chamber to ensure the airtight space of the inoculation chamber, so that the inoculator can not walk back and forth in the inoculation chamber to avoid dust flying, and the inoculator can not come in and go out of the inoculation chamber to forbid all the behaviors of large dust amount and bacteria carrying.

⑸ when inoculating, firstly opening the hundred-level laminar flow cover, inoculating personnel holding the inoculating gun with one hand, inoculating into the middle part of the fungus stick according to the fixed quantity (generally 20-50 ml), and plugging the cotton plug into the mouth of the inoculated fungus stick with the other hand to prevent the hypha from being polluted by the outside mixed fungi.

⑹ transfer of the articles from the outside to the inside is prohibited during inoculation, and non-inoculated persons must not leave the inoculation chamber without authorization from the technical responsible person and without strict dust and bacteria removal without approval from the group leader.

⑺ after inoculation, the various wastes left in the inoculation chamber need to be cleaned, the ultraviolet lamp is turned on, and the ozone device is turned on to leave the inoculation chamber.

⑻ the inoculation room equipment needs regular maintenance, the filtering net of the hundred-level laminar flow hood is cleaned once a month, the filtering sponge of the ozone machine is replaced once a month, and the space sterility degree is detected regularly to ensure the stable production.

Mycelium culture:

⑴ sterilizing the bacteria-cultivating room, cleaning the ceiling, wall and floor with 1:100 washing powder, sucking water with mop or mop cloth, drying the bacteria-cultivating room (25-30 deg.C) for 3 days, and fumigating with sterilizing agent such as BIJIESU and GUBAO for 2 days.

⑵ when the fungus sticks enter the fungus culturing room, the fungus sticks are prepared into warehouse entry records according to the time of entering the fungus culturing room, the sterilization time, the inoculators, the seed production personnel and the fungus specification in equal batches and hung on the fungus culturing frame.

⑶ fungus culturing rack should be placed neatly and stably, the clamping grooves between the upper and lower layers should be aligned and clamped to ensure stability and no shaking, and a 50cm or so passageway is left between the two rows of racks to facilitate the patrol of technicians and the transportation of fresh air.

⑷ the temperature of the bacteria culture room and the concentration of CO2 are strictly controlled, the black fungus belongs to saprophytic aerobic fungus, the temperature is controlled to be 26-28 ℃ in the first 3 days after inoculation, the temperature is controlled to be optimally 22-26 ℃ in the rest time, the temperature is controlled to be not more than 32 ℃ at most and not less than 6 ℃ at least, the concentration of CO2 is controlled to be not more than 1000PPM (the black fungus enters the bacteria culture room without feeling stuffiness) by a timing ventilation device, and the air filtering membrane is replaced once a month.

⑸ inspecting the bacteria-cultivating rooms at 9:00, 13:00, 17:00 and 21:00 every day, recording the temperature, humidity, CO2 concentration, hypha growth speed, hypha pollution and cleaning condition of each bacteria-cultivating room, checking whether the air-conditioning and ventilation facilities are normal, filling in "the bacteria-cultivating room inspection record table", wherein the inspection is generally performed by 2 persons, and the safety is required when climbing the shelf.

⑹ when the fungus sticks are delivered to the field for management, the delivery time, purchasers, purchase quantity, technical information of each batch of fungus sticks production, etc. are filled in the record table of fungus stick delivery, and each buyer and company is responsible for the management.

⑺ the fungus sticks are evacuated, and the fungus culturing room is sterilized and disinfected in time according to the first requirement, and the second batch of fungus sticks are prepared for fungus culturing in the storehouse.

⑻ when power failure or air conditioner ventilation failure occurs, the diesel engine power generation system should be started in time, which is important to ensure the ventilation of the bacteria-culturing room, when the temperature of the bacteria-culturing room exceeds 30 ℃, the air conditioners in each bacteria-culturing room are supplied with power one by one in a circulating interval, and the temperature is controlled in a safe range.

56. Field management of mulberry branches and agarics in a greenhouse;

⑴ transportation

Loading:

vertical loading: the frame is easy to deform and release, the layer height is generally required to be reduced as much as possible, and the loading and unloading are convenient and the production efficiency is high.

Horizontal loading: the device is good in placement, time-consuming and labor-consuming, but is light in deformation and wall separation, large in loading capacity up to more than ten layers, and easy to get hot in long-distance transportation.

And (3) transportation: the fungus bags can not be seen when being heated in a short time, clear water is spitted during later management, yellow water or hyphae become light and become gray hyphae which sometimes occurs later. The opening, hanging bag and budding can not be seen. Therefore, the long-distance transportation and loading of the black fungus bags are carried out at the lowest temperature, and one of the important reasons for ensuring that the fungus bags are not required to produce transverse warps, remove walls, crush and the black fungus is produced stably and has high yield when the fungus bags deform is ensured.

⑵ placing fungus sticks in greenhouse

The ground of the greenhouse is leveled, after the rope is hung, the height of each time, each space and several times of swinging can be accurately calculated, after how many bags can be hung on each greenhouse, the greenhouse can be directly put into the greenhouse, and if the old fungus greenhouse needs to be disinfected in advance.

Paving ground cover, mulching film, straw curtain and other heat-insulating, cool-insulating and clean materials.

If the fungus bags in the shed are well-matured after physiology, the bags are deformed and easily separated from the wall, can be scattered and stacked. If the temperature of the greenhouse is higher (more than 30 ℃), the dispersion is better. The stack is better when the air temperature is below 25 ℃. The deformed and detached bag can be returned to the original and is easy to be heated, but the later opening and bag hanging work efficiency is high, and the common bag hanging is mostly arranged in a way of swinging.

The color of the fungus bag is not white when the fungus bag is transported. The method is characterized in that the air-borne hyphae fall down under the influence of external force when the fungus bags are transported, the fungus bags can be recovered after being placed in a greenhouse for 3-5 days at proper temperature and humidity, the temperature is recovered faster when the temperature is 25-30 ℃, otherwise, the temperature is slower, and the greenhouse is shaded and cooled if the temperature of the greenhouse exceeds 30 ℃ or the temperature between the fungus bags is 30 ℃ when the fungus bags are laid. Ventilating and cooling in the morning and evening or spraying mist water for cooling.

After a period of management, the physiological after-ripening can whiten the thallus, and the fungus bag has elasticity and can be opened without being very hard. The air temperature is low when the fungus bags enter the greenhouse, the placing time is long, otherwise, the placing time is short, and the placing time is short for middle and early-maturing varieties.

⑶ opening of fungus bag

The number of the openings is about 120-. The second eye will typically be made 3-7 days after the first eye.

The shape of the opening is a straight line shape, a nail eye shape and a Y shape. The shape of the fungus is favorable and the technical management is slightly difficult; y-shaped ears are easy to emerge, more ears are produced during the emergence period, and the yield is high. The flower type is more vegetables, and the root of the edible fungus is large; a font is between the two.

The opening size can be punctured in principle, the depth is generally required to be 3-5MM, the length is 3-6MM, the opening is basically 3MM, the opening is an electric machine at present, the fungus bags are different in curing degree, the culture material particles are different in size, the shape, size and depth of the opening are different, the germination of the agaric is slow when the opening is too deep, the root of the big-leaved agaric in the mouth is large, and the agaric is easy to fall when the mouth is shallow.

⑷ management after opening

The height of the opened pendulum layer is 6 layers at most, 4-5 layers are formed as much as possible, the distance between the lap and the lap is as large as possible (about 10 CM), and a large amount of biological heat can be generated 3-5 days after the opening, namely, the lap can be burned by the lap, and the lap is also a key point after the opening.

Temperature and humidity management after opening: keeping the temperature at 20-30 ℃ and the humidity at 75-90%, and absolutely preventing rain in the first three days after opening. Compared with a sun-shading net, the ventilation in the morning and evening can not be reduced, and the ventilation cooling or the scattered stack cooling can be enhanced only by utilizing the low temperature at night in advance.

After 3-10 days, the temperature and humidity are suitable for 3-5 days, opening hypha is closed (namely the opening is white, the bag wall is not grown, the fungus bag can be hung as if the fungus bag does not have an opening, if the opening is not uniformly developed, the upper layer is developed well, the lower layer is not developed well, the bag is poured once, the uppermost layer of the poured bag is placed at the lowermost layer, the lowermost layer is placed at the uppermost layer, the temperature is high, the sealing is quick, and the sealing is slow at low temperature.

⑸ hanging bag

1. When the greenhouse is tied in a rope, the ventilation of the greenhouse is determined according to experience, and the bag can be hung in a high terrain with a higher density. The distance between each string is 20-30cm, the distance between each trip is 28-35cm, the middle operation channel is 75-80cm, if an air channel is reserved in the middle of the shed length, each string is 7-8 rods, and the bottom is 20-30cm away from the ground, but not lower than 20 cm. If the greenhouse is in a low-lying or densely-forested environment, the density of hanging the bags close to the mountain depends on the small density and a plurality of ventilation channels are reserved. Ventilating ducts are usually reserved every 15M or so, namely two ropes are hung at each time longitudinally.

2. The difficulty of hanging the bags in the greenhouse is to solve the problem of ventilation, the better the ventilation is, and the yield and the quality are good.

3. The rope buckle of the hanging bag needs to be tied to be hard, the hanging bag is firm, and people can pass back and forth between the hanging bags without falling the bags.

⑹ accelerating germination and growing ear

1. After the bag is hung in the greenhouse, sunlight is directly irradiated for one day, and ultraviolet rays in the sunlight are utilized to kill a part of mixed bacteria. Because the opening to the hanging bag is different from the opening in the early stage for 3 to 10 days, a plurality of mixed bacteria can be infected in the mouth and the bag wall, and the drying of the bag for one day has a plurality of advantages. Generally, a plastic film and a sunshade net are rolled up, and if the watering is carried out continuously in rainy days or cloudy days, the watering can be directly carried out after a bag is hung.

2. Promoting the generation of ear bud at 15-30 deg.C, preferably 20-25 deg.C, 75-95% humidity, and 85-95% humidity. The germination is best at the most suitable temperature and humidity, the germination is fast, the germination is neat, and the method is one of key points for ensuring stable yield and high yield, if the temperature is lower than 20 ℃, particularly below 15 ℃ for a long time, the germination of the agaric is easy to bulge, the agaric bud grows without being exported, and the agaric grows on the wall of the bag. And later-period temperature is high. Because the ear buds in the bag wall are expanded, the bag wall is irrigated to cause water accumulation, and the yield is reduced. The main management during this period is below 15 degrees or above 30 degrees. Without watering, the red root and the mildew of the agaric are easy to be caused.

3. Watering is carried out in a way that less watering and frequent watering are carried out, the watering size is judged according to the temperature and the humidity, or the face fungus bag is touched and attached by hands to feel water drops. If the watering time of the spray water is 3-10 minutes longer and the water spraying time is 1-5 minutes like light rain, the watering time interval is generally 30-120 minutes, and the watering can be controlled regularly on the premise that the water cannot be watered too much.

4. The shed film and the sunshade net are adjusted in time in the daytime and at night according to the temperature and the humidity, and the shed film and the sunshade net are rolled up or put down.

5. Generally, the sprouts can grow after 7-15 days of management. The transformation process of sprouting is sealing white hypha, forming black eye circles at the mouth shape, growing the rice grain size primordium and growing the rice grains to be about 1cm, and the process is managed by stages.

a. In the black eye period, a small black circle is formed at the cut, watering is carried out less time, and the frequent watering of the atomized water is generally maintained for 5-10 minutes.

b. In the primordial growth period, namely, the agaric grows to the size of rice grains to the size of soybeans, namely, the agaric buds grow out of the seal, the watering is noticed not to be too large, the agaric buds are generally large after being seen to be transparent, mellow and soft, the agaric buds are large after being touched by hands, the agaric buds are hard and elastic, if the water is watered, ventilation or air drying is needed, and the greenhouse film and the sunshade net are rolled up for 1-3 days after the water is removed.

c. In the normal growth period of the agaric, when the agaric grows to be larger than 1cm, the fingernail should be watered with large water, the agaric can grow without continuously watering once, and a plurality of dry-wet alternate processes are required. Dry means that the agaric is dried in the sun and dried to the root of the agaric, the water is generally stopped for 2 to 3 days, and then the water is poured. Wet means that the edible fungus is watered thoroughly. When the agaric is dry, watering needs to be conducted thoroughly for many times, the watering is not excessive, and the agaric can break the agaric by how much water is fed into a water tower. The key point is that the agaric is watered to a good degree, and the agaric is generally watered to be full and crystalline or is watered by kneading and layering the agaric with hands. In the growth stage, when most of the fungus grows to 2-5cm, certain strong light irradiation is needed to thicken and blacken the fungus, the quality of the fungus is improved, and certain sun shading is needed to ensure that a layer of villi is arranged on the back surface of the fungus, namely, the back and the ventral surfaces (the front and the back surfaces) of the fungus are two colors when the fungus drains water, the ventral surface is black, and the point that the back surface is gray is also an important mark of the black fungus grade standard.

⑺ picking up

1. Most of agaric can be picked up when the agaric grows to 7cm above 3cm, if the agaric is uneven due to various reasons, the agaric can be picked up when the agaric grows to 5cm above, water is cut off before picking, the agaric is generally checked for the degree of water drainage, and the agaric can be picked up when the edge of the agaric is rolled up or the abdomen and the back of the agaric are in two colors. The color of the dried agaric picked up after being dried can be different if water is cut off, the shape and the color of the dried agaric are not good, the water is generally cut off for more than 2 hours, but the agaric cannot be dried in the sun.

2. The principle of picking the agaric is to pick the agaric with larger size and smaller size, generally, the agaric with smaller opening is picked with larger size, the agaric with larger opening is sprouted with more opening, and the agaric with thicker length is picked with smaller size. The size of the collected agaric is related to the variety, the back of the medium-early-maturing agaric has no diameter or a small diameter, but the yield is low, the quality is good, and the quality is high; the back of the middle-late maturing agaric variety needs to be adopted with a larger diameter or a plurality of diameter ear pieces, and the shape of the middle-late maturing agaric variety is not good after the middle-late maturing agaric variety is adopted with a smaller diameter and is dried in the sun; generally, the yield of the medium-maturing variety is a little, but none of them is absolute.

⑻ air-drying

The dry and wet degree of the agaric is observed, and the agaric is dried slightly and is dried slightly to be thicker; slightly wet and thinner. The agaric cannot be dried quickly when being aired, the current agaric is opposite to the prior airing method, the agaric is dried in one day in the past, and the agaric is light in weight. Now the thicker the ear the better. Drying for 5-7 days allows the ear edge and ear root to shrink simultaneously, and the black fungus is small and the black oil is mostly dried out.

⑼ post-harvest management

Stopping water for 2-3 days after picking once, normally managing, controlling thirst, watering, ventilating and enhancing illumination for 3-5 days after drying ear buds, and finishing picking after the ear is circularly picked for 5-7 days for the second time.

By detecting several black fungus products, the selenium content of the mulberry branch black fungus is 0.17mg/kg, the selenium content of the wild oak black fungus is 0.097mg/kg, and the selenium content of the common black fungus is 0.10mg/kg, which are respectively higher than 75% and 70%. The calcium content of the mulberry branch black fungus is 1.026g/kg, and the calcium content of the common black fungus is 0.504g/kg, which is improved by more than one time. Specifically, the results are shown in Table 1. The technology improves the nutrition and health care value of the black fungus and accelerates the innovation of high-quality edible fungus resources.

TABLE 1 comparison of the nutritional ingredients of three Auricularia auricula-judae

The above description is only a preferred embodiment of the present invention, and the scope of the present invention is not limited thereto, and any simple modifications or equivalent substitutions of the technical solutions that can be obviously obtained by those skilled in the art within the technical scope of the present invention are within the scope of the present invention.

Claims (5)

1. A method for culturing black fungus by utilizing mulberry branches is characterized by comprising the following steps:

step one, selection of a cultivation environment: the cultivation field is selected to have good ecological condition, flat terrain, clean surrounding environment, no pollution source and good water quality, and reaches the national sanitary standard;

step two, mulberry branch crushing and material preparation: selecting dry and mildew-free mulberry twigs with bark, pulverizing into 0.2-0.8cm mulberry twig powder with a pulverizer, or directly pulverizing fresh mulberry twigs, and drying in the sun for use to prevent mildew during storage;

step two, cultivation season: the inoculation time can be generally 3-4 months;

step three, variety selection: selecting a variety which is suitable for local weather characteristics, high in yield, high in quality, strong in stress resistance and strong in market competitiveness;

step four, the formula of the culture medium: 87-89 parts of mulberry twig chips, 9-11 parts of bran, 0.8-1.2 parts of gypsum, 0.8-1.2 parts of lime and 50-55 parts of water;

step five, cultivation

5.1 preparation of the fungus stick

5.2. And (3) sterilization: performing normal-pressure steam sterilization, stacking the material rods in a shape like a Chinese character 'jing' or a bottle pile, reserving gaps of 1-2 cm between the material rods, reserving gaps for every 3-5 layers to facilitate steam circulation, keeping the central material temperature to be more than 98 ℃ within 4-5 hours after beating cold air, and keeping for 12-14 hours to ensure the sterilization effect;

5.3, taking out of the pot and cooling: cleaning, killing insects and sterilizing the cooling field in advance; after sterilization, naturally cooling to below 50-60 ℃ and discharging from a stove, wherein the raw material bag is easy to burst when the temperature is too high; lightly taking and lightly placing the material rods out of the stove, and moving the material rods into a sterilized cooling field to be cooled to natural temperature;

5.4. inoculation: before inoculation, the inoculation box is disinfected by aerosol disinfectant, and the surfaces of both hands and instruments are disinfected by 75% alcohol; cooling the material bag to below 28 ℃, selecting a blocking strain to be inoculated and slightly higher than the surface of the hole, and then sleeving a bag and tying the bag;

5.5. mycelium culture: suspension type fungus bag cultivation

The growth of the fungus hyphae is greatly influenced by the culture temperature and humidity, the fungus hyphae need to be distinguished and mastered according to different growth periods for normal growth of the fungus hyphae, and the fungus bags are inoculated and then moved into a culture field for suspension type stacking; the whole culture process is roughly divided into three stages: germination period, growth period, and maturation period; checking regularly, and turning the pile every 10-15 days; under normal conditions, the fungus sticks grow over white hyphae from 45 days to 60 days;

5.6. ear emergence management;

step six, harvesting

When the ear is eight-nine percent mature, the ear stem becomes thin, the ear lobe is spread and slightly droops, a small amount of spores are generated on the compound surface, the picking can be carried out when dew is not dry and the ear is in a damp and soft state in sunny days, impurities are removed, and the ear is dried in the sun;

seventhly, clearing the field after the mining

Cleaning impurities after harvesting, stopping spraying water for 5-7 days to promote hypha recovery, starting spraying fine water on 8 days to keep ear sticks moist, and spraying water again to grow out of ears after new ear buds are formed.

2. The method for cultivating black fungus by using mulberry twigs according to claim 1, wherein in the third step, the variety of black fungus is selected from one of black fungus 916, black wei 5 or shennong 7.

3. The method for culturing black fungus by using mulberry twigs according to claim 1, wherein the step 5.1 is specifically as follows: pre-wetting the main material, uniformly mixing with bran, gypsum and lime, adding water while stirring, fully stirring, bagging within 5 hours after stirring, respectively packaging into bags with 15cm × 55cm polyethylene bags, fastening the bag openings, inspecting the walls of the bacteria bags, and ensuring no thinning, abrasion or puncture.

4. The method for culturing black fungus by using mulberry twigs according to claim 1, wherein in the step 5.5, the germination period is as follows: controlling the temperature to be 26-28 ℃ before 10 days after inoculation, controlling the room temperature to be about 22-24 ℃ after 10-20 days, and frequently checking and paying attention to the stack temperature, increasing the stack temperature and increasing the ventilation;

and (3) growing period: after 20 days of inoculation, the temperature is preferably controlled to be about 22 ℃, when the white mycelium grows to the diameter of 10cm-15cm around the inoculation hole, an outer layer of a bag is removed to increase oxygen and ventilate, the growth of the mycelium is promoted, meanwhile, pile turning is carried out again after 10-15 days, and the fungus sticks infected with the mixed fungi are eliminated;

and (3) mature period: after 35-40 days after inoculation, the mycelium gradually enters a physiological maturation stage, the room temperature is preferably controlled at 20-22 ℃, ventilation adjustment is mainly performed by opening and closing doors and windows, and bacteria burning caused by overhigh temperature is avoided; when white protruding buds appear after mycelia grow over the bag, cutting 2cm openings at two ends of the stick to promote the development of sporophore; at this time, the sesame can enter the sesame shed.

5. The method for culturing black fungus by using mulberry twigs according to claim 1, wherein the step 5.6 specifically comprises the following steps: (1) selecting an earfield: the method has the advantages that the method is good in wind gating, sufficient in sunlight, convenient in water source, free of pollution source, capable of preventing waterlogging and used as an earfield;

(2) and (3) ear field construction: in a field ear field, drainage ditches with the depth of 30cm are arranged at the periphery of the field ear field, a ridge bed is shaped like a turtle back, the width is 140-150 cm, the length is not limited, and supports with 3 rows in the longitudinal direction, the height is 30-35 cm and the line spacing is 40-50 cm are erected by small wood (bamboo) rods or iron wires; after disinfecting and killing insects, the ridge-up bed is paved with hay or straws and a sunshade net, and a micro-spray pipe is installed;

(3) puncturing and ear-stimulating: puncturing holes on a sunny day with the temperature below 25 ℃, puncturing 150-180 ear outlet holes with the depth of 1-1.5 cm around fungus sticks by using a special puncturing machine for sterilized black fungus, stacking the fungus sticks after puncturing according to a Chinese character jing shape or a triangle shape, reducing the stacking amount in unit area, and simultaneously strengthening ventilation management; controlling the temperature below 25 ℃ and the humidity at about 80% during the fungus culturing period after puncturing, covering a plastic film for heat preservation when the humidity is lower than 20 ℃, ventilating every day, and accelerating the puncturing to culture the fungus for 7-10 days, recovering the mycelium of the puncturing, and discharging the mycelium out of the field;

(4) arranging the fields: generally, in the middle 10 th month to the middle 11 th month, clear or cloudy weather is selected for the arrangement; filling water to moisten the ear field one day before the drainage; when the mushroom sticks are arranged on a field, the mushroom sticks lean against the support at an angle of 60-70 degrees with the ground and are uniformly distributed, and the distance between the mushroom sticks and the support is 10-15 cm; 7000 to 8000 bars are arranged per mu in a field ear field;

(5) and (3) ear field management: the principle of 'alternation of wetting and drying' is adopted, less spraying is carried out in rainy days, and more spraying is carried out in sunny days and dry days; or the lug is in a leather shape and is sprayed more, and the lug is in a blister shape and is sprayed less or not sprayed; after the fungus sticks are arranged, the fungus sticks are kept from losing water, a small amount of fine water is sprayed for many times, and the relative humidity is kept between 80% and 90%; after young ears are formed and grow into small ears, water is sprayed for 1-2 times every day, and the water spraying amount is increased along with the growth of the ears so as to promote the growth of the young ears; the water spraying amount in the earning period is correspondingly increased along with the growth of the fruiting body, and the dry-wet difference is enlarged, namely dry days and wet days, the dry-wet difference alternately promotes the growth, and the relative humidity is kept between 90% and 95%; if continuous rainy days occur, covering measures are needed; if the water is accumulated in the bar inner connecting sheet and the bag, the bag is taken out and the ear is produced.