CN109258306A - A kind of elegant precious mushroom strain mating system rapidly and efficiently - Google Patents
A kind of elegant precious mushroom strain mating system rapidly and efficiently Download PDFInfo
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- CN109258306A CN109258306A CN201811356949.XA CN201811356949A CN109258306A CN 109258306 A CN109258306 A CN 109258306A CN 201811356949 A CN201811356949 A CN 201811356949A CN 109258306 A CN109258306 A CN 109258306A
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G18/00—Cultivation of mushrooms
- A01G18/40—Cultivation of spawn
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G18/00—Cultivation of mushrooms
- A01G18/20—Culture media, e.g. compost
Abstract
The present invention provides a kind of elegant precious mushroom strain mating system rapidly and efficiently, the step of this method uses are as follows: (1) extraction of elegant precious mushroom parent species: culture is carried out to elegant precious mushroom fructification tissue and obtains parent species；(2) parent species ultrasonic pulse is handled；(3) expansion of parent species is numerous；(4) parent species parent species Liquid Culture: are obtained into seed liquor by shaking flask culture and seed tank culture；(5) seed liquor fermentation tank culture: seed liquor is seeded in fermentor, and elegant precious mushroom liquid spawn can be made by stirring aerobic fementation.The present invention changes the solid breeding method of traditional elegant precious mushroom strain, has used microbial liquid fermentation technique in the breeding of elegant precious mushroom strain, has prepared good elegant precious mushroom liquid spawn.The method Spawn incubation period provided by the invention is short, biomass is big, breeding is high-efficient, and the strain of acquisition is pure, without miscellaneous bacteria, and fruiting is neat after inoculation, convenient for management；And method and process of the invention is simple, save the cost, and some fermentation raw material can also gather materials on the spot, waste utilization.
Description
Technical field
The invention belongs to Edible Fungi fields, and in particular to a kind of elegant precious mushroom strain mating system rapidly and efficiently.
Background technique
Edible mushroom refers to the gill fungus bacterium (macro fungi) that fructification is very large, eats for the mankind, is commonly referred to as mushroom.According to statistics
The edible mushroom of state is up to 938 kinds, more than 50 of artificial cultivation.Edible mushroom protein usually all rich in and amino acid,
Content is several times to tens times of general vegetables and fruit, and multivitamin and microelement and lower are also contained in edible mushroom
Fat content, be a kind of delicious flavour, healthy food full of nutrition, be widely loved by the people.Elegant precious mushroom (Pleurotus geesteranus) it is a kind of good edible mushroom, scientific name handle mushroom belongs to Eumycota, basidiomycetes in taxonomy
Guiding principle, Agaricales, Pleurotaceae, Pleurotus.Elegant precious mushroom is the gill fungus bacterium of subtropical and tropical zones, originates in India, for many years with China
The phoenix-tail mushroom of cultivation belongs to of the same race, but economical character and commodity property have many differences.Elegant precious mushroom one derives from Taiwan, by
Handsome small in its mushroom body, stem is 5 ~ 6 centimetres long, and for bacteria cover diameter less than 3 centimetres, flavor is extremely delicious, and hence obtain one's name elegant treasure.20th century
The 90's Mos, Fujian, Shanghai, the success of zhejiang and other places elegant precious mushroom introducing and planting, into 21st century, since consumption demand increases,
Elegant precious mushroom cultivation scale expands rapidly.
Elegant precious mushroom is not only full of nutrition, but also delicious flavour, and protein content is higher than Dual Mushroom mushroom, mushroom, straw mushroom, connects
Nearly meat, 3 ~ 6 times higher than general vegetables, quality is delicate, and fiber content is few；Elegant precious mushroom contains 17 kinds with upper amino acid, more may be used
Expensive to be, it, which contains human body itself, to manufacture, and threonine, lysine, the homo amino acid etc. usually lacked in food, there is " mushroom
The title of middle superfine product ".Elegant precious mushroom is as other edible mushrooms, containing various bioactivators, such as macromolecule polysaccharide, β ~ glucose
With RNA complex, nucleic acid degradation product, triterpene compound etc., there is anticancer, give protection against cancer, improve the immunity of the human body and other effects, also
Antibacterial, antiviral, lowering blood pressure and blood fat, it is hypoglycemic the effects of, therefore elegant precious mushroom have huge market Development volue, development
Potentiality are very big.
With the fast development of mushroom industry, the yield and sales volume of elegant precious mushroom increasingly expand.It is more in order to meet
The market demand, the cultivation mode of elegant precious mushroom gradually develop to intensive scale cultivation and facility by the dispersion cultivation of each peasant household
Change cultivation, cultivation season is also by autumn and winter and spring and summer develop simultaneously till now annually cultivating of original autumn and winter.And strain is
The prerequisite of elegant precious mushroom cultivation, the quality of strain, yield, production cycle be restrict that elegant precious mushroom cultivates on a large scale it is important because
Element.The strain used of elegant precious mushroom cultivation at present is mostly that farmer or plantation factory are obtained after multistage expansion is numerous by parent species, between batch
Differ greatly, inoculum concentration difference it is big, and tend to have living contaminants, inoculation operation is not easy to grasp.The production of traditional elegant precious mushroom is generally adopted
Inoculated and cultured is carried out with solid spawn, the cultivation cycle of solid spawn is long, and preparation process is cumbersome, breeds inefficiency；Solid bacterium
The incubation of kind needs biggish space, and need to put into a large amount of human and material resources, and required cost of material is also high；Work as solid
After strain inoculation, since the cultural hypha period is long, culture substrate mycelium cell age differs greatly up and down, and front end mycelia is in shape of sprouting
When state, stromal surface mycelia has been approached aging, causes cultivar fruiting irregular, it is difficult to meet the production requirement of scale.
And liquid spawn is with the production of hybrid seeds period is short, inoculation efficiency is high, it is fast to colonize cover, mycelium growing period is short, cell age is consistent, dirty
The low advantages such as low with production cost of dye rate, edibility bacterium large-scale planting, it has also become the development trend of edible mashroom cultivating industry.
Liquid spawn is exactly the fermenting microbe bred out with microbial liquid fermentation technique, can be obtained in a short time using this method
A large amount of mycelium, and it is with short production cycle, and production capacity is big, and strain properties are stablized, and is convenient for industrialized production.We are elegant to influencing
Each key factor of precious mushroom liquid fermentation effect culture medium, cultivation temperature, revolving speed, ventilatory capacity, inoculum concentration for example used etc. into
It has gone further investigation, and has optimized entire culture process, be finally obtained preferable achievement, realize the quick of elegant precious mushroom strain
Efficiently breeding.
Summary of the invention
The purpose of the present invention is to provide a kind of elegant precious mushroom strain mating systems rapidly and efficiently, and microbial liquid is fermented
Technology application breeds field to elegant precious mushroom strain, so as to greatly shorten the Spawn incubation period, improves strain and breeds efficiency, be conducive to
The extensive cultivation for meeting elegant precious mushroom uses.
To achieve the above object, the present invention adopts the following technical scheme:
A kind of elegant precious mushroom strain mating system rapidly and efficiently, which is characterized in that this method uses following steps:
(1) extraction of elegant precious mushroom parent species: screening has this strain characteristic, healthy and strong disease-free, the morphologically normal mature son of elegant precious mushroom in fact
Body is placed in superclean bench and operates, and first carries out cleaning disinfection to fructification appearance with 75% ethyl alcohol, is then cut with sterile razor blade
Take the tissue block of stem and cap intersection to be connected on improvement PDA culture medium inclined-plane, be placed in 23 ~ 27 DEG C of temperature, relative humidity 75 ~
It is cultivated under 85% dark condition, mycelia starts to sprout after 1 ~ 2d, filters out the inclined-plane conduct that mycelial growth rate is fast and growing way is strong
Parent species；
(2) parent species ultrasonic pulse is handled: carrying out ultrasonic wave arteries and veins to the parent species filtered out using ultrasonic intelligent biochemical reaction instrument
Punching processing, supersonic frequency 35KHz, power 30W, ultrasonic time 15s, ultrasound interval 5s, ultrasonic whole process 40min；
(3) expansion of parent species is numerous: above-mentioned parent species being inoculated into the plate equipped with improvement PDA culture medium, are placed in identical as step (1)
Under conditions of cultivate, during which every 2d check 1 time, depollution is removed in time, until mycelia covers with plate；
(4) parent species Liquid Culture: by expand it is numerous after elegant precious mushroom parent species fungus block according to every piece of 5mm, the inoculum concentration of 5 ~ 6 pieces/bottle is inoculated with
In the 250mL shaking flask equipped with 100mL fluid nutrient medium, it is protected from light standing and is placed on 5 ~ 7d of culture acquisition bacterium on constant-temperature table for 24 hours
Liquid；Again by bacterium solution according to 4 ~ 6%(v/v) inoculum concentration be inoculated in the seeding tank equipped with same liquid culture medium, temperature 22 ~ 26
DEG C, 6 ~ 8d of stirring ventilation culture obtains seed liquor；
(5) seed liquor fermentation tank culture: fermentation medium is fitted into fermentor, and liquid amount is the 70% ~ 80% of fermentation tankage size,
Be passed through air stirring it is uniform after, sealing tank mouth carry out high pressure steam sterilization；After sterilizing, it is down to 0 to fermentor internal pressure power, training
Base temperature is supported when being down to 30 DEG C or less, aseptically according to 4 ~ 6%(v/v) inoculum concentration obtained from inoculation mouth injection step (4)
The elegant precious mushroom seed liquor obtained starts fermentation tank culture after closing inoculation mouth, and culture period indirect fermentation tank temperature degree is controlled at 23 ~ 25 DEG C,
Revolving speed control is in 140 ~ 160r/min, and in 0.01 ~ 0.03MPa, ventilatory capacity is controlled in 4.0 ~ 5.0L/min, Yi Gongpei the voltage-controlled system of tank
5 ~ 7d is supported, standard compliant fermenting microbe is obtained.
In the step (1) and (3), the ingredient of PDA culture medium is improved are as follows: 140 ~ 180g/L of potato, 55 ~ 75g/ of sweet potato
L, 15 ~ 25g/L of brown sugar, 16 ~ 20g/L of xanthan gum, 3 ~ 4g/L of protein hydrolysate, 1 ~ 2g/L of yeast extract, 18 ~ 25ml/L of brewer's wort, phosphorus
Acid dihydride 1.5 ~ 2.0g/L of potassium, 0.4 ~ 0.6g/L of magnesium sulfate, 0.03 ~ 0.05g/L of copper sulphate, surplus is distilled water, and pH is adjusted to
5.8~6.2。
In the step (4), the ingredient of fluid nutrient medium are as follows: 10 ~ 14g/L of starch from sweet potato, 12 ~ 18ml/L of corn juice, phosphorus
Acid dihydride 1.8 ~ 2.2g/L of potassium, 0.9 ~ 1.1g/L of magnesium sulfate, 3.5 ~ 4.5g/L of peanut shell powder, 2.3 ~ 2.7g/L of yeast extract, beans
Cake 2.5 ~ 3.1g/L of powder, 0.7 ~ 1.5g/L of rice bran, 1.7 ~ 2.5g/L of conch meal, 12 ~ 16mg/L of gibberellin, study on brassinolide 1.5
~ 1.9mg/L, surplus are distilled water, and pH is adjusted to 5.8 ~ 6.2.
In the step (4), at 23 ~ 27 DEG C, revolving speed is controlled in 120 ~ 140r/min, seed for the temperature control of constant-temperature table
In 80 ~ 120r/min, ventilatory capacity is controlled in 0.8 ~ 1.2L/min for the mixing speed control of tank culture.
The ingredient of fermentation medium in the step (5) are as follows: 20 ~ 26g/L of sweet potato powder, wheat stalk powder 18 ~ 24g/L are red
Sugar 5 ~ 10g/L, 4 ~ 8g/L of blackstrap, mushroom bran 2.2 ~ 3.0g/L of carbide, fermented grain 1.0 ~ 1.8g/L of grain, potassium dihydrogen phosphate 0.8 ~
1.5g/L, fish offal 2.6 ~ 3.0g/L of powder, 3.9 ~ 4.5g/L of seaweed meal, 1.5 ~ 2.7g/L of tealeaf residue, dung beetle 3.3 ~ 3.9g/L of powder,
0.5 ~ 1.0g/L of magnesium sulfate, 60 ~ 90mg/L of puridoxine hydrochloride, 1.2 ~ 1.8mg/L of stearic acid, 0.9 ~ 1.3mg/L of Tween-40,
Surplus is distilled water, and the initial pH of fermentation medium is adjusted to 6.0 ~ 6.5.
The parameter of step (5) the mesohigh steam sterilizing are as follows: 0.13 ~ 0.15Mpa of steam pressure, 121 DEG C of sterilising temp,
Sterilization time 60min.
Standard compliant fermenting microbe refers to that culture medium clear is not muddy, slightly sticky in the step (5), without miscellaneous
Bacterium, free from extraneous odour；Mycelium pellet (or mycelia segment) even suspension is not stratified in liquid, and diameter is 1.0 ~ 2.0mm.
The invention has the following advantages that
1) the method Spawn incubation period of the invention is short, and biomass is big, using traditional elegant precious mushroom solid spawn mode of reproduction, needs
It cultivates 60 ~ 70 days, and uses mode of reproduction of the invention, it is only necessary to cultivate 25 days or so, substantially increase strain breeding effect
Rate.The present invention carries out ultrasonic pulse processing to elegant precious mushroom parent species, excites the vigor of mycelia, promotes the growth of mycelia, most
Achieve the purpose that increase hypha biomass eventually.
2) the pure no miscellaneous bacteria of liquid spawn cultivated of the present invention, the speed of growth is fast, and cell age is consistent, and fruiting is neat, yield
Height may be directly applied to the extensive cultivation of elegant precious mushroom, have very high practicability and promotional value.
3) method and process of the invention is simple, and required raw material are cheap and easily-available, and some of them can also gather materials on the spot, waste
It utilizes, reduces environmental pollution, while method of the invention also reduces the consumption in human and material resources, soil, more save the cost.
Specific embodiment
In order to make the object, technical scheme and advantages of the embodiment of the invention clearer, combined with specific embodiments below to this
Invention is described further.Reagents or instruments used without specified manufacturer in embodiment, being can be by commercially available purchase
The conventional products of acquisition.
Embodiment 1
It is edible in Guannan County of Jiangsu Province modern agriculture garden according to elegant precious mushroom strain mating system rapidly and efficiently provided by the invention
It is tested bacterium production base, the specific steps are as follows:
(1) purification of elegant precious mushroom parent species: screening has this strain characteristic, healthy and strong disease-free, the morphologically normal mature son of elegant precious mushroom in fact
Body is placed in superclean bench and operates, and first carries out cleaning disinfection to fructification appearance with 75% ethyl alcohol, is then cut with sterile razor blade
The tissue block of stem and cap intersection is taken to be connected to the improvement PDA culture medium (ingredient of the culture medium are as follows: potato 160g/L, red
Potato 65g/L, brown sugar 20g/L, xanthan gum 18g/L, protein hydrolysate 3.5g/L, yeast extract 1.5g/L, brewer's wort 21.5ml/L, phosphoric acid
Potassium dihydrogen 1.8g/L, magnesium sulfate 0.5g/L, copper sulphate 0.04g/L, surplus are distilled water, and pH is adjusted on 6.0) inclined-plane, is placed in
25 DEG C of temperature, relative humidity 80% dark condition under cultivate, mycelia starts to sprout after 2 ~ 3d, and it is fast to filter out mycelial growth rate
Strong inclined-plane is as parent species with growing way.
(2) parent species ultrasonic pulse is handled: carrying out ultrasound to the parent species filtered out using ultrasonic intelligent biochemical reaction instrument
Wave impulse processing, supersonic frequency 35KHz, power 30W, ultrasonic time 15s, ultrasound interval 5s, ultrasonic whole process 40min.
(3) expansion of parent species is numerous: above-mentioned parent species being inoculated into the plate equipped with improvement PDA culture medium, step (1) phase is placed in
It is cultivated under conditions of, is during which checked 1 time every 2d, depollution is removed in time, until mycelia covers with plate.
(4) parent species Liquid Culture: by expand it is numerous after elegant precious mushroom parent species fungus block according to every piece of 5mm, the inoculum concentration of 5 ~ 6 pieces/bottle
It is inoculated in the 250mL shaking flask equipped with 100mL fluid nutrient medium, is protected from light standing and is placed on constant-temperature table for 24 hours, constant-temperature table
At 24 DEG C, revolving speed control cultivates 5 ~ 7d and obtains bacterium solution in 130r/min for temperature control；Again by bacterium solution according to 5%(v/v) inoculation
Amount is inoculated in the seeding tank equipped with same liquid culture medium, and 24 DEG C of temperature, mixing speed is controlled in 100r/min, ventilatory capacity control
System cultivates 6 ~ 8d and obtains seed liquor in 1.0L/min；The ingredient of fluid nutrient medium used are as follows: starch from sweet potato 12g/L, corn juice
16ml/L, potassium dihydrogen phosphate 2.0g/L, magnesium sulfate 1.0g/L, peanut shell powder 4.0g/L, yeast extract 2.5g/L, beancake powder
2.8g/L, rice bran 1.1g/L, conch meal 2.1g/L, gibberellin 14mg/L, study on brassinolide 1.7mg/L, surplus are distilled water,
PH is adjusted to 6.0.
(5) seed liquor fermentation tank culture: by the fermentation medium (ingredient of the culture medium are as follows: sweet potato powder 23g/L, Wheat Straw
Stalk powder 21g/L, brown sugar 7.5g/L, blackstrap 6g/L, mushroom bran carbide 2.6g/L, fermented grain grain 1.4g/L, potassium dihydrogen phosphate 1.2g/
L, fish offal powder 2.8g/L, seaweed meal 4.2g/L, tealeaf residue 2.1g/L, dung beetle powder 3.6g/L, magnesium sulfate 0.75g/L, hydrochloric acid pyrrole
Tremble alcohol 75mg/L, stearic acid 1.5mg/L, Tween-40 1.1mg/L, and surplus is distilled water, and the initial pH of fermentation medium is adjusted to
6.25) be fitted into fermentor, liquid amount be ferment tankage size 70% ~ 80%, be passed through air stirring it is uniform after, sealing tank mouth carry out
High pressure steam sterilization, the parameter of sterilizing are 0.13 ~ 0.15Mpa of steam pressure, 121 DEG C of sterilising temp, sterilization time 60min；It goes out
After bacterium, it is down to 0 to fermentor internal pressure power, when culture medium temperature is down to 30 DEG C or less, aseptically according to 5%(v/v)
The elegant precious mushroom seed liquor that is obtained from inoculation mouth injection step (4) of inoculum concentration, start fermentation tank culture after closing inoculation mouth, cultivate
At 24 DEG C, revolving speed control exists in 150r/min, the voltage-controlled system of tank in 0.02MPa, ventilatory capacity control the control of period fermentation jar temperature
4.5L/min, one co-cultures 6d, and acquisition culture medium clear is not muddy, slightly sticky, no miscellaneous bacteria, free from extraneous odour；Mycelium pellet (or bacterium
Silk segment) even suspension is not stratified in liquid, and diameter is the standard compliant fermenting microbe of 1.0 ~ 2.0mm.
Embodiment 2
The influence that ultrasonic pulse processing grows oyster mushroom filament: 1 step of embodiment (2) is cancelled, other steps and reality
Apply the measurement that example 1 is identical, to the fermenting microbe progress mycelia bulb diameter, mycelium pellet density and the dry mycelial weight that finally obtain.Bacterium
Pompon measuring diameter uses blood cell counting plate；Mycelium pellet density measurement: 1ml fermenting microbe is taken to be diluted with water 10 times, in magnifying glass
Under count number, then convert；Dry mycelial weight measurement: mycelium is collected by filtration in 100ml fermenting microbe, 60 after clear water flushing
DEG C drying to constant weight, weighs dry weight.
As seen from the results in Table 1, compared with not handling, after being handled using ultrasonic pulse, the mycelium pellet of final fermenting microbe
Diameter, mycelium pellet density and dry mycelial weight are all significantly improved, and possible cause is the ultrasonic wave using suitable intensity and frequency
Pulse handles mycelium, can excite mycelia vigor, improves mycelial growth rate and certain bioenzyme activities, reaches raising fermentation
The purpose of thallus hypha biomass.
Embodiment 3
Elegant precious mushroom mother culture media it is preferred: by the improvement PDA culture medium used in 1 step of embodiment (1) and (3) replace with
Under several conventional mediums, 1. PDA culture medium (potato 200g/L, glucose 20g/L, agar 20g/L)；2. MGYA culture medium
(peptone 1g/L, maltose 20g/L, yeast 2g/L, agar 20g/L)；3. corn flour sucrose culture medium (corn flour 40g/L, sugarcane
Sugared 10g/L, agar 20g/L), pH is adjusted to 6.0, when comparing mycelium germination time, mycelial growth potential and mycelia and covering with plate
Between.
As seen from the results in Table 2, using elegant precious mushroom mother culture media of the invention, mycelium germination time and full ware time are most
Short, mycelial growth potential is most strong, illustrates that mycelia growth is fast on mother culture media of the invention, density is big, grows vigorous.The present invention
Mother culture media nutrient abundant can be provided, be suitable for oyster mushroom pieces growth.
Embodiment 4
Elegant precious mushroom fluid nutrient medium it is preferred: by the fluid nutrient medium used in 1 step of embodiment (4) replace with it is following it is several often
Advise culture medium, culture medium 1. (potato 200g/L, glucose 20g/L, peptone 5g/L, potassium dihydrogen phosphate 2g/L, magnesium sulfate
1g/L)；Culture medium 2. (potato 120g/L, peptone 0.5g/L, soybean powder 2.5g/L, potassium dihydrogen phosphate 3g/L, magnesium sulfate
1.5g/L)；Culture medium 3. (sucrose 30g/L, corn flour 10g/L, yeast extract 10g/L, potassium dihydrogen phosphate 2g/L, magnesium sulfate 0.5g/
L), pH is adjusted to 6.0, and the measurement of mycelia bulb diameter, mycelium pellet density and dry mycelial weight is carried out to the seed liquor of acquisition.Bacterium
Pompon measuring diameter uses blood cell counting plate；Mycelium pellet density measurement: taking 1ml seed liquor to be diluted with water 10 times, under magnifying glass
Number is counted, then is converted；Dry mycelial weight measurement: mycelium is collected by filtration in seed liquor, is dried to for 60 DEG C after clear water rinses
Constant weight weighs dry weight.
As seen from the results in Table 3, using fluid nutrient medium of the invention, mycelia bulb diameter, mycelium pellet density, dry mycelial weight are all
Reach maximum value, illustrates that the growth of oyster mushroom filament is best.Fluid nutrient medium nutrient of the invention is rich and varied, carbon source,
Nitrogen concentration proportion is suitable, also added phytohormone gibberellin, study on brassinolide, can more promote mycelial growth.
Embodiment 5
Elegant precious mushroom fermentation medium it is preferred: by the fluid nutrient medium used in 1 step of embodiment (5) replace with it is following it is several often
Advise culture medium, culture medium 1. (corn flour 30g/L, sucrose 20g/L, yeast powder 3g/L, potassium dihydrogen phosphate 2g/L, magnesium sulfate 1g/
L)；Culture medium 2. (potato 200g/L, wheat bran 30g/L, peptone 30g/L, yeast powder 3g/L, potassium dihydrogen phosphate 1.5g/L, sulphur
Sour magnesium 1g/L)；3. (glucose 20g/L, yeast powder 15g/L, potassium dihydrogen phosphate 2g/L, magnesium sulfate 0.5g/L), pH is for culture medium
6.0 are adjusted to, the measurement of mycelia bulb diameter, mycelium pellet density and dry mycelial weight, measurement side are carried out to the fermenting microbe of acquisition
Method is the same as embodiment 2.
As seen from the results in Table 4, using fermentation medium of the invention, mycelia bulb diameter, mycelium pellet density, dry mycelial weight are all
Reach maximum value, the growth of oyster mushroom filament is best during illustrating fermentation tank culture.Fermentation medium of the invention is in addition to adding
Add properly mixed carbon source and nitrogen source, also added some discarded raw material full of nutrition such as wheat stalk powder, blackstrap, bacterium
Chaff carbide, fermented grain grain, fish offal powder, seaweed meal, tealeaf residue, dung beetle powder etc., can waste utilization and promote it is mycelial
Growth.
The influence that revolving speed grows oyster mushroom filament in fermentation tank culture condition: by fermentation training in 1 step of embodiment (5)
During supporting, fermentor revolving speed is set as 120 r/min, 150 r/min, 180 r/min, carries out mycelia to the fermenting microbe of acquisition
The measurement of bulb diameter, mycelium pellet density and dry mycelial weight.
As seen from the results in Table 5, when revolving speed is 150r/min, the mycelium pellet density and dry mycelial weight of elegant precious mushroom fermenting microbe
Reach peak, and mycelia bulb diameter is moderate；When revolving speed is 120r/min, although mycelium pellet is relatively large in diameter, mycelium pellet density
Lower, dry mycelial weight is lower；When revolving speed is 180r/min, although mycelium pellet density improves, high speed oscillation will affect mycelia
Ball forms certain diameter, and mycelia has the phenomenon that also self-dissolving, and dry mycelial weight can also be reduced.
The influence that ventilatory capacity grows oyster mushroom filament in fermentation tank culture condition: by fermentation in 1 step of embodiment (5)
During culture, ventilatory capacity is set as 2.5L/min, 4.5L/min, 6.5L/min, and it is straight to carry out mycelium pellet to the fermenting microbe of acquisition
The measurement of diameter, mycelium pellet density and dry mycelial weight.
As seen from the results in Table 6, when ventilatory capacity is 4.5L/min, mycelia bulb diameter, the mycelium pellet of elegant precious mushroom fermenting microbe
Density and dry mycelial weight reach peak；Under the conditions of 2.5L/min mycelia bulb diameter, mycelium pellet density and dry mycelial weight all compared with
It is low, illustrate that ventilatory capacity is lower and affect oxygen content, and then affects mycelial growth；Mycelia bulb diameter under the conditions of 6.5L/min
It is not much different, but mycelium pellet density is lower, dry mycelial weight is also accordingly reduced, therefore 4.5L/min is than convenient ventilatory capacity.
The influence that temperature grows oyster mushroom filament in fermentation tank culture condition: by fermentation training in 1 step of embodiment (5)
During supporting, fermentation jar temperature is set as 21 DEG C, 24 DEG C, 27 DEG C, and it is close to carry out mycelia bulb diameter, mycelium pellet to the fermenting microbe of acquisition
The measurement of degree and dry mycelial weight.
As seen from the results in Table 7, between 22 ~ 28 DEG C, oyster mushroom filament can normal growth, but mycelium pellet is straight at 24 DEG C
Diameter, mycelium pellet density, dry mycelial weight all reach maximum value, therefore are the optimum temperature of elegant precious mushroom fermentation tank culture.
Embodiment 6
Conventionally prepare elegant precious mushroom solid spawn, the specific steps are as follows:
(1) produce parent species: same screening has this strain characteristic, healthy and strong disease-free, morphologically normal elegant precious mushroom mature sporophore, uses
75% ethyl alcohol cuts stem after sterilizing to fructification and the tissue block of cap intersection is connected on PDA culture medium inclined-plane, is placed in temperature
25 DEG C, cultivate under the dark condition of relative humidity 80%.
(2) original seed is produced: after the mycelia of parent species covers with medium slant, 1 ~ 1.5cm of picking2Fungus block be inoculated into and be equipped with
In the original seeds bottle of pedigree seed culture medium, the formula of pedigree seed culture medium uses weed tree sawdust 72%, wheat bran 18%, corn flour 9%, gypsum 1%,
Water is added to turn uniformly by 60% water content again after mixing, pedigree seed culture medium loading amount is 85% or so of original seeds bottle volume, is placed in temperature
26 DEG C, cultivate under the dark condition of relative humidity 70%.
(3) it produces cultivar: after mycelia covers with original seeds bottle, the mycelia of original seeds bottle surface aging being removed, original seed is connect
In kind to cultivation bag culture medium, the formula of bag culture medium is cultivated using weed tree sawdust 78%, wheat bran 20%, white sugar 1%, gypsum 1%, is mixed
Water is added to turn uniformly by 60% water content again afterwards, inoculum concentration is advisable with very thin culture underglass bag surface, is placed in 26 DEG C of temperature, phase
To being cultivated under the dark condition of humidity 70%.
Embodiment 7
The fermenting microbe that embodiment 1 obtains and the solid spawn that embodiment 5 obtains are cultivated according to elegant precious mushroom conventional cultivation method,
Specific step is as follows:
(1) preparation elegant precious mushroom cultivation culture medium: according to conventional formulation cotton seed hulls 40%, bagasse 40%, wheat skin 18%, lightweight carbon
Sour calcium 2% prepares culture medium, adds water to turn uniformly by 60 ~ 65% water content again after mixing, culture medium is distributed into polybag, often
Packed wet feed 2.5kg/L, is converted into siccative 1.0kg/L, puts on seal mouth ring, and ring internal plug cotton carries out high pressure sterilization.
(2) it is inoculated with bacterium germination: until culture medium temperature is down to room temperature in bag, fermenting microbe and reality that embodiment 1 is obtained
The solid spawn for applying the acquisition of example 4 is inoculated on culture medium, is placed in bacterium germination culture in culturing room, continues to cultivate after mycelia purseful
3d, the control of culture room temperature at 22 ~ 24 DEG C, irradiate 65 ~ 75% without light by relative humidity control.
(3) management of producing mushroom: moving into mushroom house culture for the culture medium of above-mentioned mycelia purseful, and mushroom house temperature is controlled at 18 ~ 22 DEG C,
Relative humidity is controlled, and a small amount of natural scattered light irradiation is given once daily 85 ~ 95% during sporophore growth, when cap is long extremely
2cm or so, lid edge are involute, stem is extended to 5cm or so, and spore can harvest the first damp mushroom before not yet launching.
As seen from the results in Table 7, using elegant precious mushroom fermenting microbe of the invention, the average mycelia purseful time is 18d, is averaged out
The mushroom time (number of days of the first damp mushroom fruiting is seeded to since strain) is 32d, and uses the average mycelia purseful of solid spawn
Time is 33d, and average fruiting time is 49d, illustrates that the elegant precious mushroom fermenting microbe speed of growth of the invention is significantly higher than solid bacterium
Kind, and mycelium growth vigor is vigorous after strain inoculation, and pollution rate is also extremely low, and anti-miscellaneous bacteria ability is significant, the average production of the first damp mushroom
Amount reaches 0.762 kg/ bag, and the average product than use solid spawn improves 0.112 kg/ bags, therefore of the invention quick
Efficient elegant precious mushroom strain mating system has great promotional value.
Embodiments described above is a part of the embodiment of the present invention, instead of all the embodiments.Reality of the invention
The detailed description for applying example is not intended to limit the range of claimed invention, but is merely representative of selected implementation of the invention
Example.Based on the embodiments of the present invention, obtained by those of ordinary skill in the art without making creative efforts
Every other embodiment, shall fall within the protection scope of the present invention.
Claims (7)
1. a kind of elegant precious mushroom strain mating system rapidly and efficiently, which is characterized in that this method uses following steps:
(1) extraction of elegant precious mushroom parent species: screening has this strain characteristic, healthy and strong disease-free, the morphologically normal mature son of elegant precious mushroom in fact
Body is placed in superclean bench and operates, and first carries out cleaning disinfection to fructification appearance with 75% ethyl alcohol, is then cut with sterile razor blade
Take the tissue block of stem and cap intersection to be connected on improvement PDA culture medium inclined-plane, be placed in 23 ~ 27 DEG C of temperature, relative humidity 75 ~
It is cultivated under 85% dark condition, mycelia starts to sprout after 1 ~ 2d, filters out the inclined-plane conduct that mycelial growth rate is fast and growing way is strong
Parent species；
(2) parent species ultrasonic pulse is handled: carrying out ultrasonic wave arteries and veins to the parent species filtered out using ultrasonic intelligent biochemical reaction instrument
Punching processing, supersonic frequency 35KHz, power 30W, ultrasonic time 15s, ultrasound interval 5s, ultrasonic whole process 40min；
(3) expansion of parent species is numerous: above-mentioned parent species being inoculated into the plate equipped with improvement PDA culture medium, are placed in identical as step (1)
Under conditions of cultivate, during which every 2d check 1 time, depollution is removed in time, until mycelia covers with plate；
(4) parent species Liquid Culture: by expand it is numerous after elegant precious mushroom parent species fungus block according to every piece of 5mm, the inoculum concentration of 5 ~ 6 pieces/bottle is inoculated with
In the 250mL shaking flask equipped with 100mL fluid nutrient medium, it is protected from light standing and is placed on 5 ~ 7d of culture acquisition bacterium on constant-temperature table for 24 hours
Liquid；Again by bacterium solution according to 4 ~ 6%(v/v) inoculum concentration be inoculated in the seeding tank equipped with same liquid culture medium, temperature 22 ~ 26
DEG C, 6 ~ 8d of stirring ventilation culture obtains seed liquor；
(5) seed liquor fermentation tank culture: fermentation medium is fitted into fermentor, and liquid amount is the 70% ~ 80% of fermentation tankage size,
Be passed through air stirring it is uniform after, sealing tank mouth carry out high pressure steam sterilization；After sterilizing, it is down to 0 to fermentor internal pressure power, training
Base temperature is supported when being down to 30 DEG C or less, aseptically according to 4 ~ 6%(v/v) inoculum concentration obtained from inoculation mouth injection step (4)
The elegant precious mushroom seed liquor obtained starts fermentation tank culture after closing inoculation mouth, and culture period indirect fermentation tank temperature degree is controlled at 23 ~ 25 DEG C,
Revolving speed control is in 140 ~ 160r/min, and in 0.01 ~ 0.03MPa, ventilatory capacity is controlled in 4.0 ~ 5.0L/min, Yi Gongpei the voltage-controlled system of tank
5 ~ 7d is supported, standard compliant fermenting microbe is obtained.
2. a kind of elegant precious mushroom strain mating system rapidly and efficiently according to claim 1, which is characterized in that the step
(1) and in (3), the ingredient of PDA culture medium is improved are as follows: 140 ~ 180g/L of potato, 55 ~ 75g/L of sweet potato, 15 ~ 25g/L of brown sugar,
16 ~ 20g/L of xanthan gum, 3 ~ 4g/L of protein hydrolysate, 1 ~ 2g/L of yeast extract, 18 ~ 25ml/L of brewer's wort, potassium dihydrogen phosphate 1.5 ~
2.0g/L, 0.4 ~ 0.6g/L of magnesium sulfate, 0.03 ~ 0.05g/L of copper sulphate, surplus are distilled water, and pH is adjusted to 5.8 ~ 6.2.
3. a kind of elegant precious mushroom strain mating system rapidly and efficiently according to claim 1, which is characterized in that the step
(4) in, the ingredient of fluid nutrient medium are as follows: 10 ~ 14g/L of starch from sweet potato, 12 ~ 18ml/L of corn juice, 1.8 ~ 2.2g/ of potassium dihydrogen phosphate
L, 0.9 ~ 1.1g/L of magnesium sulfate, 3.5 ~ 4.5g/L of peanut shell powder, 2.3 ~ 2.7g/L of yeast extract, 2.5 ~ 3.1g/L of beancake powder, rice
0.7 ~ 1.5g/L of chaff, 1.7 ~ 2.5g/L of conch meal, 12 ~ 16mg/L of gibberellin, 1.5 ~ 1.9mg/L of study on brassinolide, surplus are to steam
Distilled water, pH are adjusted to 5.8 ~ 6.2.
4. a kind of elegant precious mushroom strain mating system rapidly and efficiently according to claim 1, which is characterized in that the step
(4) in, at 23 ~ 27 DEG C, revolving speed is controlled in 120 ~ 140r/min, the mixing speed of seed tank culture for the temperature control of constant-temperature table
Control is controlled in 80 ~ 120r/min, ventilatory capacity in 0.8 ~ 1.2L/min.
5. a kind of elegant precious mushroom strain mating system rapidly and efficiently according to claim 1, which is characterized in that the step
(5) ingredient of fermentation medium in are as follows: 20 ~ 26g/L of sweet potato powder, 18 ~ 24g/L of wheat stalk powder, 5 ~ 10g/L of brown sugar, blackstrap 4
~ 8g/L, mushroom bran 2.2 ~ 3.0g/L of carbide, fermented grain 1.0 ~ 1.8g/L of grain, 0.8 ~ 1.5g/L of potassium dihydrogen phosphate, fish offal powder 2.6 ~
3.0g/L, 3.9 ~ 4.5g/L of seaweed meal, 1.5 ~ 2.7g/L of tealeaf residue, dung beetle 3.3 ~ 3.9g/L of powder, 0.5 ~ 1.0g/L of magnesium sulfate, salt
Sour 60 ~ 90mg/L of pyridoxol, 1.2 ~ 1.8mg/L of stearic acid, 0.9 ~ 1.3mg/L of Tween-40, surplus are distilled water, fermentation training
It supports the initial pH of base and is adjusted to 6.0 ~ 6.5.
6. a kind of elegant precious mushroom strain mating system rapidly and efficiently according to claim 1, which is characterized in that the step
(5) parameter of mesohigh steam sterilizing are as follows: 0.13 ~ 0.15Mpa of steam pressure, 121 DEG C of sterilising temp, sterilization time 60min.
7. a kind of elegant precious mushroom strain mating system rapidly and efficiently according to claim 1, which is characterized in that the step
(5) standard compliant fermenting microbe refers to that culture medium clear is not muddy, slightly sticky in, no miscellaneous bacteria, free from extraneous odour；Mycelium pellet
(or mycelia segment) even suspension is not stratified in liquid, and diameter is 1.0 ~ 2.0mm.
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| CN112481138A (en) * | 2020-12-07 | 2021-03-12 | 贵州大学 | Preparation method for high-yield polysaccharide based on sweet potato wastewater submerged fermentation |
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