CN109845586A - A kind of flammulina velutipes liquid strains preparation method of the growing point of mycelia containing high quantity - Google Patents
A kind of flammulina velutipes liquid strains preparation method of the growing point of mycelia containing high quantity Download PDFInfo
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- CN109845586A CN109845586A CN201910117778.3A CN201910117778A CN109845586A CN 109845586 A CN109845586 A CN 109845586A CN 201910117778 A CN201910117778 A CN 201910117778A CN 109845586 A CN109845586 A CN 109845586A
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- mycelia
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- growing point
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Abstract
The invention discloses a kind of flammulina velutipes liquid strains preparation methods of growing point of mycelia containing high quantity.Its method are as follows: one, by raw material of powdered compost matrix prepare needle mushroom solid spawn as inoculum；Two, the strain of preparation is smashed to pieces as powdered strain, seeding tank of the access equipped with fluid nutrient medium.The inoculum that the present invention is produced using pulverulent solids strain as flammulina velutipes liquid strains, preparation method is simple, facilitates and can use for a long time；Seed tank culture uses special agitating mode, it accesses after powdered strain first using high speed of agitator, with high efficiency dispersion solid spawn, increase growing point, then stop stirring can electric power saving consumption, then high-speed stirred increases significantly mycelia growing point, cultivates 6 h finally with activity recovery late stage of culture in short-term sufficiently to smash mycelia；Using preceding tune seed liquor pH to 8-9, inhibit varied bacteria growing after being conducive to inoculation.Flammulina velutipes liquid strains are prepared with great application prospect using the method for the present invention.
Description
Technical field
The invention belongs to edible mushroom and application of biological engineering fields, are related to a kind of needle mushroom of growing point of mycelia containing high quantity
Liquid spawn preparation method.
Background technique
Needle mushroom (Flammulina velutipes) it is also known as dried mushroom, structure bacterium etc., it is a kind of famous edible in the world
Bacterium, in the cultivation extensively of the country such as China, Japan and South Korea.Needle mushroom delicious flavour, it is full of nutrition, it is a kind of high protein, it is low
Heat, the wholefood of high polyoses content.Needle mushroom contains 18 kinds of amino acid, and wherein the content of arginine, lysine is higher than normal
See edible mushroom, therefore, needle mushroom has good facilitation to teen-age growth and development, there is the title of " increasing intelligence mushroom ".In addition, golden
Needle mushroom also has higher medical value, such as reduces cholesterol, promotes gastrointestinal peristalsis, antitumor, strengthen immunity physiological activity,
Therefore, needle mushroom is a kind of edible mushroom type with high economic value, and development potentiality is huge.Currently, China is in the world
Needle mushroom output and the most country of export volume.
Needle mushroom is the edible mushroom for using batch production planting technology earliest, and output is big.Currently, golden mushroom plantation is mainly adopted
It is inoculated with wood chip spawn, but wood chip spawn has that long preparation period, occupied space are big, labour's consumption is more and are not easy to automation connects
The disadvantages of kind.Therefore, the inevitable choice for being used to cultivate be from now on using other new mushroom-seed culturings, to improve production efficiency.With sawdust
Kind compare, edible fungi liquid strain have with short production cycle, inoculation is convenient, material feeding is fast, can automated production, labor intensity it is low etc.
Advantage, therefore, edible fungi liquid strain start for edible fungus culturing.In recent years, with edible fungi liquid strain production technology
Mature and cultivating facility updates and improves, and liquid spawn starts to be widely used in industrialized cultivation for needle mushroom, achieve significantly at
Effect is inexorable trend from now on using liquid strain cultivation needle mushroom.
Flammulina velutipes liquid strains preparation mainly includes following 3 steps: the one, activation of inclined-plane parent species；Two, shaking flask culture,
Activated spawn is seeded to triangle shake bottle Liquid Culture, normally about 4-7 d optionally can be further as level liquid kind
Shaking flask expands culture, is used subsequently to seeding tank inoculation；Three, seeding tank expands culture.It can be directly used for producing after seed tank culture
Cultivation.In terms of economy and pollution risk, the preparation process of aforesaid liquid strain is quite cumbersome, the period usually require 10 days with
On, in addition, preparation and incubation easily pollute.Still further aspect, it is poly- that when needle mushroom Liquid Culture, primarily forms mycelia
Collective and mycelium pellet, traditional seed tank culture mainly use slow-speed of revolution agitating mode to be cultivated, liquid bacteria at the end of culture
Kind mainly exists in the form of mycelium pellet, and therefore, mycelia growing point is few when which accesses culture material, usually needs to increase
Inoculum concentration, and be inoculated with inconvenient and be unfavorable for being dispersed in compost, therefore, develop the new or improved needle mushroom liquid of use
Process for preparing strain thereof has great importance.
Summary of the invention
The purpose of the present invention is to provide a kind of flammulina velutipes liquid strains preparation methods of growing point of mycelia containing high quantity.
A kind of flammulina velutipes liquid strains preparation method of the growing point of mycelia containing high quantity, comprising the following steps:
(1) needle mushroom solid spawn is prepared with powdered compost matrix:
(2) Liquid Culture: with sterile glass rod by solid under the needle mushroom solid spawn aseptic condition that step (1) is prepared
Strain is smashed to pieces, and pulverulent solids strain is become, and immediately or 4 DEG C after preservation 1 month, is seeded in seeding tank and is cultivated.
Step (1) method for preparing needle mushroom solid spawn is as follows:
Compost is prepared: each component in compost being uniformly mixed, after adjusting water content to 50-60%, weighs 150-200 g wet feed
Loaded in 500 mL triangular flasks, rubber plug is sealed；
Sterilizing: the compost in triangular flask is sterilized in 121 DEG C 2 h in autoclave；
Inoculation and culture: accessing activated needle mushroom slant strains to compost surface under aseptic condition after compost is cooling,
25 DEG C, cultivate under the dark situation of relative humidity 60-70%, after mycelia material feeding is complete；
Compost constituent included: 80-90 parts of wood dust, 8-18 parts of the bean powder, 2 parts of soya-bean oil after 20 meshes.
The method of step (2) described seed tank culture are as follows:
(1) seeding tank sterilizes: using 100 L seeding tanks, 121 DEG C of skies disappear 30 min；It is packed into 70 L fluid nutrient mediums, 121 DEG C of realities
Disappear 30 min；
(2) seeding tank is inoculated with: accessing the pulverulent solids strain after 200 g are smashed to pieces after culture medium is cooling；
(3) condition of culture is 25 DEG C of temperature, ventilatory capacity 0.5-1 vvm；
(4) stirring and cultural method: the 1st day 300-400 rev/min, to disperse the pulverulent solids strain of access, subsequent 2-4
It is only ventilated and does not stir, and power consumption is saved, and setting revolving speed is 300-400 rev/min at the 5th day, and 6 h utilize Strong shear power
Disperse and smash mycelium pellet and mycelium, increases mycelia growing point；Then stop stirring, continuing 6 h of culture makes mycelia restore to live
The growing point of mycelia containing high quantity is prepared using preceding with 1 sterilized moL/L NaOH tune culture solution pH to 8-9 in power
Flammulina velutipes liquid strains.
Further, the flammulina velutipes liquid strains preparation method of the growing point of mycelia containing high quantity is applied to golden mushroom plantation
In.
The application is the following steps are included: access cultivation package, every packaging for liquid spawn between aseptic inoculation using inoculating gun
1000 grams of doses weight in wet base, every packet accesses 20 mL liquid strains；Cultivation package after inoculation is transferred to culture room, 25 DEG C of dark cultures, bacterium bag
After material feeding is complete, it is transferred to mushroom room, irritates induction fruit-body formation through low temperature, illumination, high humility.
The present invention has the advantages that
1) solid spawn is prepared using powdered compost matrix, is better than the mycelia growth for being to be conducive to increase solid spawn
Point；
2) soya-bean oil is added in compost may advantageously facilitate mycelia growth, improve solid spawn quality,
3) solid seed preparation flow is few, is conducive to avoid pollution generation；
4) solid spawn aseptic condition is easy to control, is used to that liquid spawn preparation can be effectively improved when the inoculum of liquid strain preparation
Success rate and elimination recessive pollution；
5) solid spawn can long-time preservation, can be used at any time liquid spawn production, improve seed production efficiency；
6) using especially stirring strategy when seed tank culture, that is, it is raw to increase mycelia with dispersing solid strain for high-speed stirred after being inoculated with
Long point；Then stop stirring, power consumption can be reduced；Before culture terminates, 6 h of high-speed stirred, to disperse mycelium pellet and mycelium,
Increase mycelia growing point, 6 subsequent h are not stirred, and to restore mycelia vigor, are obtained high activity liquid spawn, are conducive to mycelia and eat
Material；
7) liquid spawn uses the preceding NaOH solution tune pH to 8-9 with 1 sterile mol/L, is conducive to inhibit miscellaneous bacteria raw after inoculation
It is long, reduce pollution；
8) liquid spawn that the present invention is prepared between aseptic inoculation using inoculating gun accesses cultivation package, the liquid of every packet access
Body kind amount is only the 1/2 of traditional liquid strain dosage；It is good that cultivation package mycelia after inoculation walks bacterium synchronism.
Specific embodiment
The invention discloses a kind of flammulina velutipes liquid strains preparation method of growing point of mycelia containing high quantity, following example
Further describe application method of the invention, but these embodiments be only it is normative, the scope of the present invention is not constituted
Any restrictions.In addition, without departing from the spirit and scope of the invention can details to technical solution of the present invention and form into
Row modifications or substitutions, but these modifications and replacement are still in the scope of the present invention.
Embodiment 1
1. preparing powdered needle mushroom solid spawn: sawdust being crushed, 20 meshes are crossed；
2. 80 parts of wood dust, 18 parts of bean powder and 2 parts of soya-bean oil, mixing adjust water content to 50%, 200 g of wet feed are loaded on 500
ML triangular flask, is slightly compacted, rubber plug sealing；
3. 121 DEG C of compost 2 h of sterilizing, it is ensured that sterilizing is thorough；
4. 1 piece of agar block of about 2 cm long, 1 cm wide that cut from needle mushroom inclined-plane parent species are accessed after cooling under aseptic condition
Strain is to compost surface, 25 DEG C, cultivate in the dark situation of relative humidity 60-70%, can be used as seed after mycelia material feeding is complete
Tank produces inoculum when liquid spawn；
5. seeding tank sterilizes: using 100 L seeding tanks, 121 DEG C of skies disappear 30 min；70 L fluid nutrient mediums (formula are as follows: Portugal is added
Grape sugar 20 g/L, 5 g/L of corn flour, 2 g/L of soya-bean oil, 1.0 g/L of potassium dihydrogen phosphate, 0.5 g/L of magnesium sulfate), 121 DEG C of realities
Disappear 30 min；
6. the inoculation of seeding tank: the pulverulent solids strain after 200 grams of access is smashed to pieces after culture medium is cooling；
7. condition of culture, 25 DEG C of temperature, ventilatory capacity is 1 vvm, stirring means are as follows: 300 rev/min(were to disperse to access in the 1st day
Solid spawn), do not stir within subsequent 2-4 days and (stir culture using ventilation), power consumption can be saved, from the 5th day turn
Speed is set as 300 rev/min(and is dispersed and shredded mycelium pellet and mycelium using Strong shear power, and 6 h are raw to dramatically increase mycelia
Long, 6 subsequent h stop stirring, and mycelia is made to rejuvenate, close with the activity before broken, are in high activity state；
8. strain uses preceding sterilized NaOH tune pH to about 8-9, inhibits varied bacteria growing after being conducive to inoculation, reduce pollution
Occur；
9. liquid spawn uses；
Liquid spawn is accessed into cultivation package (about 1000 grams of charge weight in wet base) between aseptic inoculation using inoculating gun, every 1 packet access
About 20 mL liquid spawns are compared with the liquid spawn (40 mL) of cultivation package access conventional method preparation.Cultivation after inoculation
Training packet be transferred to culture room, 25 DEG C of dark cultures are transferred to mushroom room after bacterium bag material feeding is complete, irritated through low temperature, illumination, high humility and
Induce fruit-body formation.It is about 38 days that bacterium bag, which walks the bacterium time, and uses the bacterium bag of traditional liquid strain inoculation to walk the bacterium time and be about
40 days；The first damp mushroom, biological transformation ratio 84.21% are harvested after fructification is mature, and use the liquid bacteria of conventional method preparation
Kind carries out inoculation cultivation, and biological transformation ratio 81.66% is lower than the former.
Embodiment 2
1. powdered needle mushroom solid spawn preparation is the same as embodiment 1.
2. needle mushroom solid spawn is used after preservation 1 month in 4 DEG C.
3. seeding tank size, disinfection and inoculation are the same as embodiment 1.
4. condition of culture, 25 DEG C of temperature, ventilatory capacity is 0.5 vvm, stirring condition are as follows: 400 rev/min(were to fill in the 1st day
The solid spawn of dispersion access), it does not stir within subsequent 2-4 days and (stirs culture using ventilation), electric power can be saved and disappeared
Consumption, the 5th day starting rotating speed, which is set as 400 rev/min(, to be dispersed using Strong shear power and shreds mycelium pellet and mycelium significantly to increase
Add mycelia growing point), 6 h, subsequent 6 h stopping stirring making mycelia rejuvenate, the mycelia vigor before detecting display and being crushed
It is close, it is in high activity state.
5. liquid spawn use is the same as embodiment 1.Mature fructification is harvested after fruiting, is computed biological transformation ratio (with one
Damp mushroom meter) it is 93.21%, higher than the cultivation (biological transformation ratio 88.62%) for using liquid spawn prepared by conventional method.
6. using the liquid spawn inoculation of this method preparation, in the case of obtaining similar growth and mycelia material feeding, inoculation
Amount is only 1/2 of liquid spawn prepared by conventional method.
The foregoing is merely presently preferred embodiments of the present invention, all equivalent changes done according to scope of the present invention patent with
Modification, is all covered by the present invention.
Claims (6)
1. a kind of flammulina velutipes liquid strains preparation method of the growing point of mycelia containing high quantity, which comprises the following steps:
(1) needle mushroom solid spawn is prepared with powdered compost matrix:
(2) Liquid Culture: with sterile glass rod by solid under the needle mushroom solid spawn aseptic condition that step (1) is prepared
Strain is smashed to pieces, and pulverulent solids strain is become, and immediately or 4 DEG C after preservation 1 month, is seeded in seeding tank and is cultivated.
2. a kind of flammulina velutipes liquid strains preparation method of the growing point of mycelia containing high quantity according to claim 1, feature
It is, step (1) method for preparing needle mushroom solid spawn is as follows:
Compost is prepared: each component in compost being uniformly mixed, after adjusting water content to 50-60%, weighs 150-200 g wet feed
Loaded in 500 mL triangular flasks, rubber plug is sealed；
Sterilizing: the compost in triangular flask is sterilized in 121 DEG C 2 h in autoclave；
Inoculation and culture: accessing activated needle mushroom slant strains to compost surface under aseptic condition after compost is cooling,
25 DEG C, cultivate under the dark situation of relative humidity 60-70%, after mycelia material feeding is complete.
3. a kind of flammulina velutipes liquid strains preparation method of the growing point of mycelia containing high quantity according to claim 2, feature
It is, compost constituent included: 80-90 parts of wood dust, 8-18 parts of the bean powder, 2 parts of soya-bean oil after 20 meshes.
4. a kind of flammulina velutipes liquid strains preparation method of the growing point of mycelia containing high quantity according to claim 1, feature
It is, the method for step (2) described seed tank culture are as follows:
(1) seeding tank sterilizes: using 100 L seeding tanks, 121 DEG C of skies disappear 30 min；It is packed into 70 L fluid nutrient mediums, 121 DEG C of realities
Disappear 30 min；
(2) seeding tank is inoculated with: accessing the pulverulent solids strain after 200 g are smashed to pieces after culture medium is cooling；
(3) condition of culture is 25 DEG C of temperature, ventilatory capacity 0.5-1 vvm；
(4) stirring and cultural method: the 1st day 300-400 rev/min, to disperse the pulverulent solids strain of access, subsequent 2-4
It is only ventilated and does not stir, and power consumption is saved, and setting revolving speed is 300-400 rev/min at the 5th day, and 6 h utilize Strong shear
Mycelium pellet and mycelium are dispersed and smashed to power, increases mycelia growing point；Then stop stirring, continuing 6 h of culture restores mycelia
The growing point of mycelia containing high quantity is prepared using preceding with 1 sterilized moL/L NaOH tune culture solution pH to 8-9 in vigor
Flammulina velutipes liquid strains.
5. a kind of flammulina velutipes liquid strains preparation method of the growing point of mycelia containing high quantity as described in claim 1-4 is any exists
Application in golden mushroom plantation.
6. application according to claim 5, which comprises the following steps: using inoculating gun between aseptic inoculation
Liquid spawn is accessed into cultivation package, every 1000 grams of packaging doses weight in wet base, every packet accesses 20 mL liquid strains；Cultivation package after inoculation
It is transferred to culture room, 25 DEG C of dark cultures are transferred to mushroom room after bacterium bag material feeding is complete, irritate induction through low temperature, illumination, high humility
Fruit-body formation.
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