CN101773052A - Preparation and industrial culture of aweto strain - Google Patents
Preparation and industrial culture of aweto strain Download PDFInfo
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- CN101773052A CN101773052A CN201010124418A CN201010124418A CN101773052A CN 101773052 A CN101773052 A CN 101773052A CN 201010124418 A CN201010124418 A CN 201010124418A CN 201010124418 A CN201010124418 A CN 201010124418A CN 101773052 A CN101773052 A CN 101773052A
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Abstract
The invention provides a new technological method for preparing and industrially culturing aweto strains. The method comprises the following steps of: using young silkworms of 2 years as a host of an aweto culture medium, preparing strains by separating the tissue of wild awetos, dying the young silkworms, providing biological conditions suitable for aweto growth, and culturing aweto products with the shapes of wild awetos and the similar contents of various food mass. The invention has simple and convenient operation process, high yield rate and low culture cost.
Description
Technical field
The present invention adopts pond bed and wide-mouth bottle dual mode cultivation Cordyceps sinensis, is a kind ofly to do the host with silkworm, and by dip-dye, the cultivation of bacterium liquid, the high-quality worm grass product similar of acquisition to wild cordyceps.
Background technology
Natural cordyceps is grown in the high and cold area of China western part and southwestern provinces and regions, and growing environment is special, resource-constrained.Because after the medical treatment of Cordyceps sinensis and the important value that the health centre has were admitted both at home and abroad, demand constantly increased.But, add Chinese caterpillar fungus producing region ecotope severe exacerbation, and artificial stealing excessively dug because natural resources quantity is few, cause the wild Chinese caterpillar fungus resource quantity to fall sharply, output constantly descends, and disparities between supply and demand are increasingly sharpened, cause medicine food raw materials market supply and demand to disconnect, cost an arm and a leg.In recent years, domestic have a lot of Chinese caterpillar fungus cultivations family all at experimental culture Cordyceps sinensis, but because the particularity of Cordyceps sinensis growth, many cultivations family is all because of many reason cultivation failures, even the cultivation that has success, but also because of the deformity of growing, reason such as the short and small and survival rate of fruit body is low, cause cultivating the cost height, can't carry out the batch production commerial growing.
The present invention is intended to by on the small-sized cultivation basis that changes original Cordyceps sinensis, through cultivation experiments for many years, the another suitable batch production commerial growing process similar to wild cordyceps, because each side environmental factor such as compost, bacterial classification, temp. and humidity, pH value, illumination and nutrition cause cultivating survival rate to reach more than 92% more near the wild cordyceps growth conditions.
Summary of the invention
Aweto strain preparation of the present invention and factory culture, it mainly is to adopt natural wild cordyceps stroma to prepare provenance through separate tissue, adopt the parasitism of the young silkworm in 2 ages as Chinese caterpillar fungus culture medium, add required various organic nutrients and the inorganic nutrients of suitable Cordyceps sinensis growth, at the ecotope that simulates the wild cordyceps producing region, the temperature that promptly adapts, humidity, under the condition of illuminance and pH value, cultivate and obtain and the wild cordyceps shape, the worm grass resources that medicine edible material from soybeans content is similar, its order ground are to provide tame Chinese Cordyceps sinensis product for the edible market of domestic and international medicine.Its method is:
1, culture device
The present invention adopts bedstead formula and two kinds of planting types of container, and these two kinds of planting types all are adapted in the indoor and plastic tunnel.The indoor 4 layers of bedstead of solid that are provided with, length is made by oneself by the real area of indoor or booth, is welded into wide 1 meter framework with three angle steel of 5cm * 5cm, and the bedstead upper strata is provided with several long 1.4 meters, wide 0.8 meter Xiao Chi bed, fix with compressive plate in the bed of pond, semifixed with wooden slats around the bed of pond, the pond height of bed is 14cm, so that frame can take off, the Chinese caterpillar fungus of conveniently gathering is provided with three-dimensional 4 layers, and every interlayer is every 0.8 meter.
2, cultivating soil preparation
(1) prescription, wild cordyceps are grown in theropencedrymion on the ground more, and last row upon row of trees has pine, China fir, oak, Chinese scholartree, elm class seeds, and there are shrub layer and herbaceous plant in lower floor, and loosing soil is moistening, and grogs is thinner, and pH value is at 5-7.In order to reach the growth conditions that adapts with wild cordyceps, the present invention adopts sandy loam 50% under the free of contamination forest land, weed tree sawdust 10%, and paddy sugar 10%, weeds powder 20%, river sand 10% sieves after the mixing, removal of contamination.
(2) sterilization treatment: with potassium dihydrogen phosphate 40g, magnesium sulfate 60g, calcium chloride 40g, zinc sulphate 60g, VB
1100, the VC100 sheet, sucrose 1.5kg, yellow mealworm slurry 3kg, magnetized drinking water 1000kg, mix stirring and dissolving, be sprayed on the cultivating soil that sieves, stir, at this moment soil is 1: 0.6 with the water ratio, reach hold agglomerating, slightly the globule does not drip, and mass of soil lands to scatter and gets final product, pH value 7.2, then, by UHT sterilization machine 0.5 second moment sterilization detoxicating, to various assorted bacterium, virus, the worm's ovum kill ratio is 100%, has so not only reduced the gasification of generation when chemicals is handled and the pollution of peculiar smell, adapted to the growth of mycelia, sterilization simultaneously thoroughly, and is time saving and energy saving, for the cultivation progress has shifted to an earlier date the time.
(3) strain preparation
1. test tube strains: peeled potatoes 230g, glucose 20g, agar 20g, oats flakes 10g, water 1000ml, after being merged the concentrated 500ml of poach, be respectively charged into long 20cm, bore 2cm in vitro, charge weight is 1/3 of a test tube length, seal with cotton balls then and tie up, put into 110 ℃ of sterilization tanks, sterilize after 50 minutes, put the inclined-plane, after waiting to solidify, the wild cordyceps stroma is washed several times with aqua sterilisa more repeatedly with alcohol-pickled 5 seconds kinds, be cut into sarconeme shape with the sterilization blade, taking a morsel is inoculated in the test tube slant central segment and seals, and moves in 28 ℃ the insulating box, and half-light was cultivated 10 days down, when waiting to see the test tube media surface and asterism shape white colony occurring, can carry out test tube and move and connect.When secondary move connect mycelia and be covered with test tube after, carry out for the third time tube and hold up strong the cultivation therefrom selecting the good quality strain silk, after the rejuvenation mycelia maturation, can change the expansion cultivation strain for the third time.Because the vegetative propagation of mycelia and the aweto strain of tube and cultivation repeatedly, its female parent gene morphs easily, is embodied in the many deformities of stroma, stroma is short and small, and quality and output descend.Therefore, will regularly carry out a sexual propagation to aweto strain in normal cultivation, promptly stromatic tissue is separated or the spore separation cultivation, can effectively avoid living contaminants and spawn degeneration like this.
2. liquid culture bacteria equipment: the present invention designs one group of simple and easy installation for fermenting of three-tube type.At present, Mycophyta industrialization bulk fermentation technology mainly is to be purpose to produce cordyceps mycelia medicinal and for health care.But, industry submerged fermentation construction investment is big, complex procedures, technical conditions require high, and mainly be to produce the medicine edible mycelium, in case sterilization does not thoroughly reach living contaminants, can cause whole jar of zymotic fluid to cultivate failure, even this installation for fermenting under the situation of batch production commerial growing, also should not adopt.At this situation, in order to produce the Cordyceps sinensis liquid spawn that a large amount of high-qualitys are used for artificial cultivations within a short period of time, just must design a kind of novel fermentation device, advantage such as it has small investment, and is easy and simple to handle, with short production cycle, and the mycelia quality is good.What the present invention designed is a kind of three pipe combined type installation for fermentings, it has reformed factory culture need advance to purchase expensive large-scale fermentation equipment, and conventional used shaking table and so on device of cultivating, only need to be equipped with stingy amount air-breather electromagnetic pump, air cleaner, the flask of 3000ml, grass tube, rubber tube, bottle closure of rubber, sterilization cotton balls, the many siphunculus of bifurcated, leather hose folder, workbench, anion air sterilizer, the ultraviolet germicidal lamp of DC-AC two-use.Be connected then, the cultivation amount by every day prepares several one group to tens one group flask installation for fermentings, when aweto strain is inoculated in the liquid nutrient medium, through direct oxygen blast, play the effect that fluctuates and ventilate, can carry out the deep layer of liquid spawn and cultivate.The advantage of this device is, the one, apparatus cost is low, has replaced shaking table used in oil-free gas compressor air-breather in the fermentor device and the liquid culture and so on device, thereby has greatly reduced cost, cost only is half of shaking table, 1/15 also low than small-sized fermentation tank arrangement cost.The culture fluid total amount of injecting in this device accounts for 60% of fermentation flask capacity in addition, and its effective rate of utilization ratio shakes bottle and improves more than 3 times; The 2nd, the preparation bacterial classification time is short, and only forming from one, two, three liquid culture to cultivation strain needs 15 days, has shortened the production of hybrid seeds time greatly; The 3rd, the cell age unanimity, solid spawn cell age up and down is difficult to homogeneous, and liquid spawn is unanimous between the higher and lower levels, and cell age is short, and the hyphal development stalwartness is used for solid state cultivation, is easy to field planting, sprouts soon, grows; The 4th, inoculate easy fast, reduce and pollute; The 5th, be not subjected to power limitations since adopt be DC-AC two-use type electromagnetic pump, promptly be have a power failure, also can operate as normal.
3. the preparation of liquid culture bacteria: glucose 30g, sucrose 10g, peptone 15g, dusty yeast 2g, amino acid composite powder 10g, silkworm chrysalis slurry 10g, yellow mealworm slurry 10g, potassium dihydrogen phosphate 0.7g, magnesium sulfate 2g, calcium chloride 1.8g, zinc sulphate 2g, VB
15, VC5 sheet, defoamer 2ml, water 1800ml.After having adopted fresh and alive silkworm chrysalis and fresh and alive yellow mealworm, making beating to filter in the liquid nutrient medium of the present invention, as the required nitrogenous source component of Chinese caterpillar fungus bacterial filament growth, these two kinds of insects include unsaturated fatty acids such as rich in protein, amino acid, mineral matter, vitamin h and linoleic acid, linolenic acid, also having chitin, natural hormone and various enzyme material, is the nitrogenous source nutrient that aweto mycelium very easily absorbs.Defoamer is to produce a large amount of foams for avoiding in incubation, and influence the feeding of air and cause metabolism undesired, easily cause the Chinese caterpillar fungus bacterial filament self-dissolving, growth harm to Chinese caterpillar fungus bacterial filament is bigger, so adopt the plant sunflower oil better as defoamer effect.
4. cultivate the workshop sterilization
Inoculate preceding 2 days with soar in the workshop, wall, ceiling, ground cleaning be clean, and open the door and window ventilation, and seal door and window next day, Lysol is measured in accordance with regulations with water be diluted to thimerosal, with atomizers spray on cultivation workshop wall, ceiling and ground, in the workshop with every 20m
2In establish-the 40w ultraviolet germicidal lamp, the sterilization 3 hours.Before inoculation,, like this, cultivate workshop inner air-cleaning Du Keda more than 80,000 grades, reduce air pollution to greatest extent with exhaust fan emptying indoor ozone peculiar smell.
5. inoculation
Children silkworm 0.6kg puts into-60 purpose nylon mesh with 2 ages, be diluted to 0.5: 1 thimerosal with a kind of water miscible medical disinfectant and sterile water, this medical disinfectant belongs to neutral bactericide, nonirritant, paint having no volatile substances, to young silkworm body surface not damaged, nontoxic, and after washing several times, noresidue is 99.9% to the kill ratios of various bacterial viruses.Be sprayed on the young silkworm body surface with watering can, spray disinfectant is after 30 seconds repeatedly, with sterile water spray irrigation number time repeatedly, naturally put down in the plastic big basin of a diameter 60cm after drying, liquid culture bacteria is diluted to bacterium and 0.3: 1 concentration of water with sterile water, be sprayed on the young silkworm body surface, obturage with black plastic cloth, under half-light, cultivate, every spray-painting in 8 hours 1 time, spray-painting is 3 days continuously, in order to obtain more infection chance, put into basin after in the time of the 2nd day mulberry leaf and liquid culture bacteria being puddled, it is infected while eating, and the combination of inner and outside is infected and is had higher success rate, and cultivates after 5 days, children silkworm body surface white hypha occurs and has been in the half-dead state of dying, can move into the cultivation workshop and carry out the earth backing cultivation.
6. cultivation management: the Polypropylence Sheet of sterilization is assisted in the bed of pond, put into the cultivating soil of sterilization, soil will loosen, can not compacting, highly be 9cm, prick-45 degree caves, slope with the wooden stick of diameter 2cm, the degree of depth is 4-5cm, the 3cm of being separated by between cave and the cave, and in the cave, spray strain liquid, the young silkworm that to contaminate is then head-up put into every cave, cave and puts one, almost puts into more than 400 young silkworm in every square metre, at last to pond bed surface sprinkling liquid spawn, go up the thick sterilization cultivating soil of one deck 1.3cm again, wipe off gently with wooden rule, can not compacting, obturage with black plastic cloth again, half-light is cultivated down, regulate the room temperature 15-23 ℃ in one month, air humidity 55-70%, compost pH value 6.8,1-1.5 room temperature is transferred to 16-25 ℃ in individual month, air humidity 62-75%, behind 1 first quarter moon, the larva head begins to split, germination and growth Chinese caterpillar fungus stroma base portion also exposes native face, at this moment should take off the Polypropylence Sheet of overspread, regulate the room temperature 16-29 ℃, air humidity 80-90%, at this moment pH value is 6.5, this stage will often be opened the door and window ventilation, and to regulate illuminance be the 190-240 lux, is beneficial to the Cordyceps sinensis stroma and grows.Through 1.5 months-2 months cultivation, stroma was long to the 6-12cm height again, thick 1.4-4mm, and shape is club-like, and color is a yellowish-brown, and sporangium forms and can gather.
(3) gather: stroma is ripe to need 110-120 days to artificial cultivation Cordyceps sinensis from being inoculated into, and through the cultivation management of 3 first quarter moons to 4 month, a cultivating and growing phase promptly comes to an end.The semifixed wooden slats on pond bed four limits is taken off, push earth gently aside with moulding the system shovel, carefully Cordyceps sinensis is taken out, clean earth and mycelia with clear water then, treat that the surperficial globule disappears after, be sprayed on the Cordyceps sinensis body surface with commercially available yellow rice wine, make it softening, then that its arrangement is straight, natural air drying is the Cordyceps sinensis finished product.
6, container culturing
(1) container culturing adopts wide-mouth bottle for well, and the also available system basin of moulding is moulded tubbing, dish;
(2) compost is identical with bed pond compost;
(3) the compost bottling time is not filled out too much, and is too real, and 2/3rds places of the general bottle capacity that requires to pack into get final product, and the soil is porous helps mycelia and stroma is grown;
(4) inoculation, wooden stick 5 holes of straight bundle down in wide-mouth bottle with 2cm, the 2 ages children silkworm of inoculation after 5 days head-up put into the hole inner surface respectively spray 1 liquid bacterial classification, go up the thick sterilization cultivating soil of one deck 1.3cm again, seal cultivation under the half-light with Polypropylence Sheet, room temperature is adjusted to 16-23 ℃, and humidity is 50-60%, young silkworm is all ossify after 10 days, and the body surface mycelia infects and all covers young silkworm whole body.
(5) management, in order effectively to save the space and to make full use of rate, postvaccinal wide-mouth bottle all moves into half-light cultivation down on the bedstead, 1.5 after individual month, children silkworm head has grown the fruit body pedestal and has exposed native face, at this moment can in the middle of the Polypropylence Sheet of overspread wide-mouth bottle, prick a hole, in order to ventilation, temperature adjustment 18-26 ℃ humidity is 70-80%, cultivate 2 first quarter moons after, the Polypropylence Sheet of overspread wide-mouth bottle can be taken off, this stage should be opened door and window 5-6 hour ventilation every day, increases the refraction light application time more than 10 hours, and luminous intensity is that 190-230 reins in, temperature is transferred in being 19-31 ℃, and humidity is 80-90%.Waited to grow into 110 days-120 days, when stroma length arrives 6-12cm, thick 1.4-4mm, shape is club-like, color is a yellowish-brown, and sporangium forms and can gather, and directly digs on earth at the peripheral place of Cordyceps sinensis stroma with moulding the system shovel, take out gently, clean, soft, air-dry with the yellow rice wine spray, be the Cordyceps sinensis finished product.
The present invention realizes the overall process that Cordyceps sinensis grows under complete manually operated condition, just must possess the special ecological condition that adapts to the wild cordyceps growth, and bacterial classification is pure, prescribing adequate nutrition, temp. and humidity and illuminance, from cultivating soil, the preparation bacterial classification must be operated under aseptic state to the operation of inoculation cultivation each side, could realize the success of artificial cultivation Cordyceps sinensis finished product.
Embodiment
Aweto strain preparation and factory culture, its method is:
1, cultivating device
Be welded into long 6 meters, wide 1 meter, high 3.2 meters metal framework with three angle steel of 5cm * 5cm, three-dimensional 4 layers, every interlayer is every 0.8 meter, 4 long 1.4 meters, wide 0.8 meter Xiao Chi beds of welding on every layer of bedstead, fix with compressive plate in the bed of pond, semifixed around the bed of pond with wooden slats, pond bedside height 14cm;
2, cultivating soil preparation
Adopt free of contamination forest land lower floor sandy loam 50%, weed tree sawdust 10%, paddy sugar 10%, weeds powder 20%, river sand 10%, mixing is sieved, then with potassium dihydrogen phosphate 40g, magnesium sulfate 60g, zinc sulphate 60g, calcium chloride 40g, sucrose 1.5kg, VB
1100, VC100 sheet, yellow mealworm slurry 3kg, magnetized drinking water 1000kg, mix stirring and dissolving after, be sprayed on the cultivating soil that sieves, soil is 1: 0.6 with the water ratio, after stirring, with UHT sterilization machine 0.5 second moment sterilizing, at this moment the compost pH value is 7.2, covers completely with Polypropylence Sheet again, and is stand-by;
3, strain preparation
(1) test tube strains: peeled potatoes 230g, glucose 20g, agar 20g, oats flakes 10g, water 1000ml, be merged poach and concentrate 500ml, after the filtration, be respectively charged into long 20cm, bore 2cm in vitro, charge weight is 1/3 of a test tube, seals to tie up and puts into the autoclaving jar and heat 110 ℃, sterilizes after 50 minutes, take out, the pendulum inclined-plane solidifies; With wild cordyceps with alcohol-pickled 5 seconds, wash repeatedly several times with sterile water again, being cut into sarconeme shape with the sterilization cutter is inoculated in the middle of the test tube slant, sealing the interior half-light of insulating box of 28 ℃ of back immigrations cultivated 10 days down, when treating that white colony forms, get the high-quality mycelia, carry out secondary and three tubes are held up strong, stand-by;
(2) liquid culture bacteria preparation: get glucose 30g, sucrose 10g, peptone 15g, dusty yeast 2g, amino acid composite powder 10g, silkworm chrysalis slurry 10g, yellow mealworm slurry 10g, potassium dihydrogen phosphate 0.8g magnesium sulfate 2g, zinc sulphate 2g, calcium chloride 1.5g, VB
15, the VC5 sheet, defoamer 2ml, water 1800ml behind its mixed dissolution, seals in the triangular flask of injection 3000ml, 110 ℃ of autoclavings, take out after 50 minutes, put on the three-tube type work in combination platform, three tubes of learning from else's experience hold up strong test tube strains 2cm ferfas silk to put into triangular flask, at this moment, insert logical oxygen pump, Oxygen tube, blast pipe, probe tube, and the air disinfection filter is installed in respectively in the middle of three conduits, seal a bottle and under half-light, left standstill 48 hours, regulate 15-28 ℃ of indoor temperature, humidity is 50-60%, carries out uninterrupted oxygen supply after 2 days, cultivated 10 days, when treating to cover with white hypha in the liquid, stop oxygen supply, take off stand-by;
4, cultivation workshop sterilization
It is clean with cleaning in the workshop to inoculate preceding 2 days, open door and window and ventilate, seal door and window next day, be diluted to thimerosal by the amount of stipulating in the medicine specification with Lysol and water, kill sterilamps and open with atomizers spray wall, ceiling, ground, the whole ultraviolet rays of general simultaneously, sterilized 3 hours;
5, inoculation
Children silkworm 0.6kg puts into 60 purpose nylon mesh with 2 ages, be diluted to 0.5: 1 thimerosal with water miscible medical disinfectant and sterile water, be sprayed on the young silkworm body surface, spray disinfectant is after 30 seconds repeatedly, with sterile water spray irrigation number time repeatedly, naturally put into the plastic big basin of a diameter 60cm after drying, liquid culture bacteria is diluted to bacterium and 0.3: 1 concentration of water with sterile water, pack in the little sprayer of sterilizing, evenly be sprayed on the young silkworm body surface, obturage with black plastic cloth, half-light is cultivated down, it is 26 ℃ that room temperature transfers to, and humidity is 50-60%, every spray-painting in 8 hours 1 time, put into basin after puddling mulberry leaf and liquid culture bacteria evenly in the time of the 2nd day, continuously spray-painting is 3 days, and young silkworm body surface moves into the cultivation workshop and carries out the earth backing cultivation when slightly white hypha occurs after 5 days;
6, cultivation and management
Sterilized Polypropylence Sheet is assisted in the bed of pond, put into the cultivating soil of sterilization, thickness is 9cm, prick the oblique wave cave of-45 degree with the wooden stick of diameter 2cm, degree of depth 4-5cm, the 3cm of being separated by between cave and the cave, and in the cave spraying liquid cultivation strain, the young silkworm that to contaminate is then head-up put into the cave, every cave is put 1, sprays liquid culture bacteria again 1 time to bed surface, pond, upward answers the thick cultivating soil of one deck 1.3cm, wipe off gently with wooden rule, obturage with black plastic cloth, half-light is cultivated down again, and room temperature keeps 15-23 ℃ in 1 month, air humidity 55-70%, at this moment the pH value in the cultivating soil is 6.8; 1-1.5 room temperature transfers to 16-25 ℃ in individual month, air humidity 62-75%; Behind 1 first quarter moon, native face is exposed in the growth of Cordyceps sinensis stroma, at this moment take off the black plastic cloth of overspread, regulate the room temperature 16-29 ℃, air humidity is 80-90%, at this moment pH value is 6.5 in the cultivating soil, and this stage is often opened the door and window ventilation, and regulates illuminance, require scattered light or light, 10 hours every days of light application time, intensity of illumination is that 190-240 reins in, again through 1.5-2 month cultivation, stroma is long to 6-12cm, thick 1.4-4mm shape is club-like, and color is a yellowish-brown, can gather when sporangium forms;
7, gather
The semifixed wooden slats in pond bed four limits is taken off, push earth gently aside, carefully Cordyceps sinensis is taken out, then the mycelia of cleaning earth and lying prostrate with clear water with moulding the system shovel, after treating that the surperficial globule disappears, be sprayed on the Cordyceps sinensis body surface, make it softening with yellow rice wine, put in order then straight, natural air drying
8, container culturing
(1) cultivating soil of sterilization is packed into 2/3rds places in the wide-mouth bottle prick 5 caves down with the wooden stick of 2cm is straight in wide-mouth bottle, get 5 children silkworms in 2 ages of contaminating after 5 days and head-up put into the cave respectively, 1 liquid cultivation strain of surface sprinkling.Go up the thick sterilization cultivating soil of one deck 1.3cm again, seal with Polypropylence Sheet, half-light is cultivated down, and room temperature transfers to 16-23 ℃, and humidity is 50-60%, and young silkworm is all ossify after 10 days, and the body surface mycelia infects whole young silkworms and covers the polypide whole body;
(2) manage and gather
Postvaccinal wide-mouth bottle is cultivated after 1.5 months under half-light, when children's silkworm head grows the fruit body pedestal and exposes native face, at the Polypropylence Sheet of overspread wide-mouth bottleneck, a middle hole, temperature adjustment 18-26 ℃ pricked, humidity is 70-80%, after cultivating 2 first quarter moons, the Polypropylence Sheet of overspread wide-mouth bottle is taken off, this stage will be opened door and window 5-6 hour ventilation, set up the light scattering time more than 10 hours, luminous intensity is that 190-230 reins in, and temperature transfers to and is 19-31 ℃, and humidity is 80-90%, treat long by 110 days-120 days, stroma is long to the 6-12cm height, and during thick 1.4-4mm, shape is club-like, color is a yellowish-brown, can gather when sporangium forms; Directly dig on earth at the peripheral place of Cordyceps sinensis stroma with moulding system shovel, take out gently, clean earth, spray softeningly with yellow rice wine, put in order straightly then, natural air drying is the Cordyceps sinensis finished product that bottle is planted.
Claims (1)
1. aweto strain prepares and factory culture, it is characterized by:
Forest land lower floor sandy loam 50%, weed tree sawdust 10%, cavings 10%, weeds powder 20%, river sand 10%, potassium dihydrogen phosphate 40g, magnesium sulfate 60g, zinc sulphate 60g, calcium chloride 40g, sucrose 1.5kg, VB
1100, VC100 sheet, yellow mealworm slurry 3kg, magnetized drinking water 1000kg;
2, with UHT sterilization machine 0.5 second moment sterilizing;
3, peeled potatoes 230g, glucose 20g, agar 20g, oats flakes 10g, water 1000ml;
4, glucose 30g, sucrose 10g, peptone 15g, dusty yeast 2g, amino acid composite powder 10g, silkworm chrysalis slurry 10g, yellow mealworm slurry 10g, potassium dihydrogen phosphate 0.8g, magnesium sulfate 2g, zinc sulphate 2g, calcium chloride 1.5g, VB
15,5 of VC, defoamer 2ml, water 1800ml;
5, be diluted to 0.5: 1 thimerosal with water miscible medical disinfectant and sterile water;
6, liquid is cultivated bacterial classification and be diluted to bacterium and 0.3: 1 concentration of water with sterile water;
7, every spray-painting in 8 hours 1 time, put into basin after puddling mulberry leaf and liquid culture bacteria evenly in the time of the 2nd day, spray-painting is 3 days continuously;
8, prick the cave, slope of-45 degree with the wooden stick of diameter 2cm, degree of depth 4-5cm, the 3cm of being separated by between cave and the cave, and in the cave spraying liquid cultivation strain, the young silkworm that to contaminate is then head-up put into the cave, every cave is put 1, sprays liquid culture bacteria again 1 time to bed surface, pond, upward answers the thick sterilization cultivating soil of one deck 1.3cm;
9, Mie Jun cultivating soil 2/3rds places in the wide-mouth bottle of packing into, wooden stick 5 caves of straight bundle down in wide-mouth bottle with 2cm, get 5 children silkworms in 2 ages of contaminating after 5 days and head-up put into the cave respectively, 1 liquid cultivation strain of surface sprinkling is upward answered the thick sterilization cultivating soil of one deck 1.3cm.
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