CN110100657A - A kind of production method of Dictyophora rubrovalvata test tube stock - Google Patents
A kind of production method of Dictyophora rubrovalvata test tube stock Download PDFInfo
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- CN110100657A CN110100657A CN201910477825.5A CN201910477825A CN110100657A CN 110100657 A CN110100657 A CN 110100657A CN 201910477825 A CN201910477825 A CN 201910477825A CN 110100657 A CN110100657 A CN 110100657A
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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
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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
Abstract
The present invention provides a kind of production methods of Dictyophora rubrovalvata test tube stock, belong to technical field of edible fungi cultivation.The production method of Dictyophora rubrovalvata test tube stock the following steps are included: the water content of adjustment sawdust medium to 60~70%, sterilizing, bevel culture medium；The sawdust medium includes sawdust, wheat bran, maize flour, soy noodle, gypsum, sugar, potassium dihydrogen phosphate, magnesium sulfate and water；Sawdust includes the sawdust that the timber of broadleaf deciduous tree species is processed；Dictyophora rubrovalvata fructification tissue block be close to culture medium by be allowed tissue block, carry out Mother culture according to 3 equal points of inoculations, 3 pieces of tissue blocks in an aseptic environment.Be obviously shortened Dictyophora rubrovalvata test tube stock incubation time using the production method, general 25~30d can tube use, than traditional parent species production method, shorten more than 20 days time, and then greatly reduce tube infection rate.
Description
Technical field
The invention belongs to technical field of edible fungi cultivation, and in particular to a kind of production method of Dictyophora rubrovalvata test tube stock.
Background technique
Currently, domestic edible mushroom mother culture media generallys use PDA culture medium, PDA culture medium has production cost low, former
The advantages that material is easy to get, and preparation is easy, but there are mycelia to be only grown in media surface using PDA culture medium as mother culture media,
Not the problem of not entering culture medium growth inside.And when preparing Dictyophora rubrovalvata parent species using PDA culture medium, compared to most of other
The mycelia of edible mushroom, Dictyophora rubrovalvata strain is extremely slim and frahile, and the speed of growth is extremely slow, about 0.5~1mm/d, and it is oblique to cover with 10cm
Face needs 50~60 days, and grows 20d or so aerial hyphae and start purpling red, aging phenomenon occurs, is unfavorable for subsequent tube
Or strain expands numerous therefore current parent species production method and limits the industrialization cultivation of Dictyophora rubrovalvata from the root.
Summary of the invention
In view of this, there is culture the purpose of the present invention is to provide a kind of production method of Dictyophora rubrovalvata test tube stock
The feature that time is short and mycelia vigor is high.
A kind of production method of Dictyophora rubrovalvata test tube stock provided by the invention, comprising the following steps:
1) water content of sawdust medium is adjusted to 60~70%, sterilizing, bevel culture medium；
The sawdust medium includes the component of following mass percentage: 60~80% sawdusts, 5~20% wheat bran, 5~
20% maize flour, 1~10% soy noodle, 0.5~1.5% gypsum, 0.5~2% sugar, 0.1~0.5% potassium dihydrogen phosphate and 0.1
~0.5% magnesium sulfate；The sawdust includes the sawdust that the timber of broadleaf deciduous tree species is processed；
2) in an aseptic environment, by the Dictyophora rubrovalvata fructification of surface sterilization from stem vertical profile, in stem split surface
Middle part tissue segmentation is inoculated on a slant medium at tissue block, by every 3 pieces of tissue blocks according to the distance of 3 equal parts, allows group
It knits block and is close to culture medium, carry out Mother culture.
Preferably, the sawdust medium includes the component of following mass percentage: 65~75% sawdusts, 10~15%
Wheat bran, 8~15% maize flours, 2~5% soy noodles, 0.8~1.2% gypsum, 0.8~1.5% sugar, 0.1~0.3% di(2-ethylhexyl)phosphate
Hydrogen potassium and 0.1~0.3% magnesium sulfate.
Preferably, the sawdust medium includes the component of following mass percentage: 70% sawdust, 14.8% wheat bran,
10% maize flour, 3% soy noodle, 1% gypsum, 1% sugar, 0.1% potassium dihydrogen phosphate and 0.1% magnesium sulfate.
Preferably, the granularity of the sawdust is 5~50 mesh.
Preferably, the sterilizing methods of sawdust medium high pressure steam sterilization 30min at 0.11~0.13Mpa.
Preferably, the broadleaf deciduous tree species include one of Salicaceae, Fagaceae, Moraceae and Juglandaceae tree species or more
Kind.
Preferably, mature, the healthy fructification of the Dictyophora rubrovalvata fructification selection 7~8.
Preferably, the disinfection of Dictyophora rubrovalvata fructification is liquor potassic permanganate or sodium dichloro cyanurate solution, institute with solution
The concentration for stating liquor potassic permanganate or sodium dichloro cyanurate solution liquid is 0.05~0.1%.
Preferably, the temperature of the Mother culture is 20~25 DEG C.
Preferably, the time of the Mother culture is 25~30d.
The production method of a kind of Dictyophora rubrovalvata test tube stock provided by the invention, using sawdust medium as Dictyophora rubrovalvata test tube
Parent species culture medium, wherein broadleaf deciduous tree species sawdust is sustainable to Dictyophora rubrovalvata strain mycelia utility with higher
Nutrition is provided for mycelia growth, wheat bran and soy noodle provide nitrogen source in a short time for cooperation, and maize flour and sugar provide carbon source and stone
Cream, potassium dihydrogen phosphate and magnesium sulfate provide inorganic salts, so that mycelia is rapidly adapted to the nutrition of new environment, ensure that mycelia is vigorous, fast
The growth of speed.Parent species are prepared using Dictyophora rubrovalvata test tube stock culture medium provided by the invention, Dictyophora rubrovalvata strain covers with test tube,
The speed of growth has a clear superiority, while mycelia is pure white sturdy, energetic, and under identical condition of culture, using conventional mother
Kind culture medium PDA culture medium prepares Dictyophora rubrovalvata test tube stock, and mycelia is slim and frahile, and the slow underfill test tube of mycelial growth rate,
And mycelia has aging, wilting phenomenon to occur, and this shows Dictyophora rubrovalvata test tube stock culture medium provided by the invention than conventional PDA
Culture medium has unrivaled advantage in the preparation of Dictyophora rubrovalvata parent species.Simultaneously in tissue separation inoculation, connect using 3 points
Kind method ensure that tissue block quickly grows mycelia instead of inoculation method some in conventional production methods, while be trained using sawdust
It is abundant with nutriment to support base, the characteristics of mycelia absorption rate is good, long-acting offer nutriment, using 3 inoculation methods
Big to the nutritional requirement and problem with nutrition supply deficiency caused by incubation time length of tissue block is avoided, therefore, this
The production method that invention provides overcomes the technology barrier of Dictyophora rubrovalvata parent species preparation for a long time, to cultivate red support bamboo on a large scale
Sweet-smelling grass provides guarantee.
Detailed description of the invention
Fig. 1 is the Dictyophora rubrovalvata test tube stock aspect graph that production method culture 24d provided by the invention is obtained；
Fig. 2 is the Dictyophora rubrovalvata test tube stock aspect graph obtained using PDA culture medium culture 24d.
Specific embodiment
A kind of production method of Dictyophora rubrovalvata test tube stock provided by the invention, comprising the following steps:
1) water content of sawdust medium is adjusted to 60~70%, sterilizing, bevel culture medium；
The sawdust medium includes the component of following mass percentage: 60~80% sawdusts, 5~20% wheat bran, 5~
20% maize flour, 1~10% soy noodle, 0.5~1.5% gypsum, 0.5~2% sugar, 0.1~0.5% potassium dihydrogen phosphate and 0.1
~0.5% magnesium sulfate；The sawdust includes the sawdust that the timber of broadleaf deciduous tree species is processed；
2) in an aseptic environment, by the Dictyophora rubrovalvata fructification of surface sterilization from stem vertical profile, in stem split surface
Middle part tissue segmentation is inoculated on a slant medium at tissue block, by every 3 pieces of tissue blocks according to the distance of 3 equal parts, allows group
It knits block and is close to culture medium, carry out Mother culture.
The present invention adjusts the water content of sawdust medium to 60~70%, sterilizing, bevel culture medium.In the present invention
In, the sawdust medium includes the component of following mass percentage: 60~80% sawdusts, 5~20% wheat bran, 5~20%
Maize flour, 1~10% soy noodle, 0.5~1.5% gypsum, 0.5~2% sugar, 0.1~0.5% potassium dihydrogen phosphate and 0.1~
0.5% magnesium sulfate；Preferably include the component of following mass percentage: 65~75% sawdusts, 10~15% wheat bran, 8~15%
Maize flour, 2~5% soy noodles, 0.8~1.2% gypsum, 0.8~1.5% sugar, 0.1~0.3% potassium dihydrogen phosphate and 0.1~
0.3% magnesium sulfate, more preferably include following mass percentage component: 70% sawdust, 14.8% wheat bran, 10% maize flour,
3% soy noodle, 1% gypsum, 1% sugar, 0.1% potassium dihydrogen phosphate and 0.1% magnesium sulfate.The sawdust includes broadleaf deciduous tree species
The sawdust processed of timber.The broadleaf deciduous tree species include Betulaceae, Salicaceae, Fagaceae, Moraceae and Juglandaceae tree
One of kind is a variety of.Dictyophora rubrovalvata strain can quickly be absorbed and utilized what the temperate zone broadleaf deciduous tree species timber provided
Nutriment, and keep mycelium growth vigor sturdy quickly.And discovery Dictyophora rubrovalvata strain is studied to nutriment in tree species type
It is selective using having, such as conifer trees can not then be utilized.In order to illustrate Dictyophora rubrovalvata strain to broadleaf deciduous tree species wood
The utilization power of bits, the present invention is with the mulberry of Chinese chestnut tree, Moraceae in Betulaceae birch and Qi wood, poplar in Salicaceae, Fagaceae
It is illustrated for tree and the walnut tree of Juglandaceae, but this can not be interpreted as limiting the scope of the invention.The wood
The granularity of bits is preferably 5~50 mesh, more preferably 30 mesh.The present invention does not have special limit to the preparation method of the sawdust medium
System, using the preparation method of this field conventional medium.The sterilizing methods of the sawdust medium preferably 0.11~
High pressure steam sterilization 30min under 0.13Mpa.The pressure of sterilizing is preferably 0.12Mpa.The method of bevel culture medium is by sawdust
Culture medium is paved into the smooth inclined-plane of inclined-plane 10cm in test tube.
After obtaining slant medium, the present invention in an aseptic environment, by the Dictyophora rubrovalvata fructification of surface sterilization from stem
Locate vertical profile, stem split surface middle part tissue segmentation at tissue block, every 3 pieces of tissue blocks are inoculated according to the distance of 3 equal parts
On one slant medium, tissue block is allowed to be close to culture medium, carries out Mother culture.
In the present invention, mature, the healthy fructification of the Dictyophora rubrovalvata fructification preferably 7~8.7~8 mature sons are in fact
Body is according to the micro-judgments of the veteran dictyophora phalloidea poultry feeders in this field.
In the present invention, the disinfection of Dictyophora rubrovalvata fructification is preferably liquor potassic permanganate or sodium dichloro cyanurate with solution
The concentration of solution, the liquor potassic permanganate or sodium dichloro cyanurate solution is 0.05~0.1%；The disinfection of the solution is soaked
Steeping the time is preferably 1~2min, more preferably 1.5min.
In the present invention, the specification of the tissue block is 5~7mm × 7~10mm.Tissue block is allowed to be close to the method for culture medium
Tissue block is gently pressed with transfer needle, makes the tissue block firm contact to culture medium, to guarantee tissue block on culture medium
It sprouts.
In the present invention, the temperature of the Mother culture is preferably 20~25 DEG C, and more preferably 23 DEG C.The Mother culture
Time be preferably 25~30d, more preferably 27~28d.By Mother culture process, tissue block is rapidly on sawdust medium
Sprouting grows mycelia, and due to containing sawdust in sawdust medium, nutrition abundant can be provided for mycelia, is conducive to mycelia and gos deep into
Nutriment is fully absorbed inside culture medium, is shortened the Mother culture time, is reduced pollution probability, while culture is obtained into parent species
Tube Shi Suozhuan test tube does not infect or pollutes substantially very low (5% or less about), shortens than the conventional method preparation dictyophora phalloidea parent species time
1 half times (needing 60 days or so), while having than the dictyophora phalloidea parent species tube infection rate (reaching 30-50%) of conventional method preparation
It significantly reduces.
It is detailed to a kind of production method progress of Dictyophora rubrovalvata test tube stock provided by the invention below with reference to embodiment
Illustrate, but they cannot be interpreted as limiting the scope of the present invention.
Embodiment 1
1, the preparation and sterilizing of sawdust medium
Be formulated as follows: poplar sawdust 70%, 14.8% wheat bran, 10% maize flour, 3% soy noodle, 1% gypsum, it is 1% sugared,
0.1% potassium dihydrogen phosphate, 0.1% magnesium sulfate.
Said components are mixed according to the proportion, the water content of culture medium is adjusted to 65%, is in vitro made into the oblique of 10cm
Face.
The culture medium of bevel is used into high pressure steam sterilization, keeps 30 under the conditions of 0.13Mpa after excluding cold air
Minute, sterilizing finishes, and it is spare to take out cooling.
2,3 inocalation methods
Mature, the healthy dictyophora phalloidea fructification of selection 7, after handling 1.5min with disinfecting solution of potassium permanganate, in ultra-clean work
Internal stem part at two valves, then is divided into that 7mm × 10mm's is small by the vaccinating lancet of the interior sterilizing of platform by fructification vertical profile in length and breadth
Square cannot cut through tissue of the fructification in addition to stem, gently provoke stem tissue block, allow each tissue block to loosen, separate, use tweezer
Tissue block is inoculated on culture medium by son or transfer needle, and every test tube classifies in three categories point, connects 3 pieces of tissue blocks, compacting, allows tissue block
It is close to culture medium.
3, Mother culture
The slant medium for being inoculated with tissue block is cultivated in 25 DEG C of incubator, observes hypha growth condition daily,
The test tube polluted is taken out.The parent species that mycelia covers with inclined-plane are subjected to tube culture, count germination rate and the sense of tube
Dye rate.
Embodiment 2
1, the preparation and sterilizing of sawdust medium
Be formulated it is as follows: walnut sawdust 63.4%, 20% wheat bran, 5% maize flour, 10% soy noodle, 0.5% gypsum,
0.5% sugar, 0.1% potassium dihydrogen phosphate, 0.5% magnesium sulfate.
Said components are mixed according to the proportion, the water content of culture medium is adjusted to 60%, is in vitro made into the oblique of 10cm
Face.
The culture medium of bevel is used into high pressure steam sterilization, keeps 30 under the conditions of 0.11Mpa after excluding cold air
Minute, sterilizing finishes, and it is spare to take out cooling.
2,3 inocalation methods
Mature, the healthy dictyophora phalloidea fructification of selection 8, after handling 1min with sodium dichloro cyanurate solution disinfection, ultra-clean
Fructification vertical profile is divided into length and breadth at two valves, then by internal stem part with the vaccinating lancet of sterilizing on workbench (or inoculating hood)
The small cube of 5mm × 7mm cannot cut through tissue of the fructification in addition to stem, gently provoke stem tissue block, make each tissue block loose
It is dynamic, separate, tissue block is inoculated on culture medium with tweezers or transfer needle, every test tube classifies in three categories point, connects 3 pieces of tissue blocks, pressure
It is real, allow tissue block to be close to culture medium.
3, Mother culture
The slant medium for being inoculated with tissue block is cultivated in 22-25 DEG C of incubator, mycelia is observed daily and grows feelings
Condition takes out the test tube polluted.
Embodiment 3
1, the preparation and sterilizing of sawdust medium
Be formulated it is as follows: Chinese chestnut wood sawdust 80%, 5% wheat bran, 11.4% maize flour, 1% soy noodle, 1.5% gypsum,
0.5% sugar, 0.5% potassium dihydrogen phosphate, 0.1% magnesium sulfate.
Said components are mixed according to the proportion, the water content of culture medium is adjusted to 70%, is in vitro made into the oblique of 10cm
Face.
The culture medium of bevel is used into high pressure steam sterilization, keeps 30 under the conditions of 0.12Mpa after excluding cold air
Minute, sterilizing finishes, and it is spare to take out cooling.
2,3 inocalation methods
The dictyophora phalloidea fructification that 7-8 is mature, healthy is selected, after handling 1min with disinfecting solution of potassium permanganate, in ultra-clean work
Fructification vertical profile is divided into 7mm at two valves, then by internal stem part in length and breadth with the vaccinating lancet of sterilizing on platform (or inoculating hood)
The small cube of × 7mm cannot cut through tissue of the fructification in addition to stem, gently provoke stem tissue block, each tissue block is allowed to loosen,
It separating, tissue block is inoculated on culture medium with tweezers or transfer needle, every test tube classifies in three categories point, 3 pieces of tissue blocks are connect, compacting,
Tissue block is allowed to be close to culture medium.
3, Mother culture
The slant medium for being inoculated with tissue block is cultivated in 23 DEG C of incubator, observes hypha growth condition daily,
The test tube polluted is taken out.
The Dictyophora rubrovalvata Mother culture 25-30d of Examples 1 to 3 preparation, mycelia cover with inclined-plane, and the parent species mycelia of preparation is clean
It is white, it is sturdy, it grows fine, vigor is relatively strong (see Fig. 1).When the parent species of culture are carried out tube, germination rate is higher, reaches 100%,
Infection rate is lower, reaches 1% or less
Comparative example 1
Prepare PDA culture medium, by PDA culture medium at 0.12Mpa high pressure steam sterilization 30 minutes, sterilizing finishes, take out.
Culture medium pendulum inclined-plane after sterilizing is obtained into PDA slant medium after cooling.
Dictyophora rubrovalvata strain is seeded on PDA slant medium according to one point method according to the method for embodiment 5, every kind of training
Support base weight reinoculation 3 in parallel.It is cultivated in 25 DEG C of incubator after inoculation, observes hypha growth condition daily, by pollution
Inclined-plane or the inclined-plane that do not sprout are rejected.Statistics success ratio of inoculation simultaneously photographs to record hypha growth condition.
Mother culture needs 60 days or so to cover with inclined-plane.But mycelia is slim and frahile, and wilting phenomenon is presented, vigor is lower (see figure
2), when the subsequent culture to tube, germination rate is low, and infection rate is higher.
Comparative example 2
PDA culture medium is prepared according to the method for comparative example 1, inclined-plane is put after sterilizing, obtains PDA slant medium after cooling.
It is seeded in PDA slant medium according to 3 inocalation methods recorded in embodiment 1, in 25 DEG C of culture after inoculation
It is cultivated in case, observes hypha growth condition daily, the inclined-plane of pollution or the inclined-plane that do not sprout are rejected.
As a result, it has been found that there is wilting phenomenon in mycelia, and mycelia color is slightly turned to be yellow when cultivating 15d or so.Analyzing reason may
It is that PDA culture medium nutritional ingredient is limited, mycelia utilizes poor problem to absorption of nutrient ingredients in PDA culture medium in addition, out
Existing incubation nutrition supply is insufficient, causes parent species vigor lower, and when the subsequent culture to tube, germination rate is low.
Comparative example 3
With poplar sawdust in Dictyophora rubrovalvata Cultivar culture medium in pine sawdust alternative embodiment 1, other operations are according to reality
Apply the method operation of the record of example 1.
The matrix such as pine, China fir preparation Cultivar culture medium cannot grow Dictyophora rubrovalvata mycelia, this illustrates pine, China fir etc.
Matrix is not suitable for doing the matrix of Dictyophora rubrovalvata culture medium.
The above is only a preferred embodiment of the present invention, it is noted that for the ordinary skill people of the art
For member, various improvements and modifications may be made without departing from the principle of the present invention, these improvements and modifications are also answered
It is considered as protection scope of the present invention.
Claims (10)
1. a kind of production method of Dictyophora rubrovalvata test tube stock, which comprises the following steps:
1) water content of sawdust medium is adjusted to 60~70%, sterilizing, bevel culture medium；
The sawdust medium includes the component of following mass percentage: 60~80% sawdusts, 5~20% wheat bran, 5~20%
Maize flour, 1~10% soy noodle, 0.5~1.5% gypsum, 0.5~2% sugar, 0.1~0.5% potassium dihydrogen phosphate, 0.1~
0.5% magnesium sulfate；The sawdust includes the sawdust that the timber of broadleaf deciduous tree species is processed；
2) in an aseptic environment, by the Dictyophora rubrovalvata fructification of surface sterilization from stem vertical profile, at the middle part of stem split surface
Tissue segmentation is inoculated on a slant medium at tissue block, by every 3 pieces of tissue blocks according to the distance of 3 equal parts, allows tissue block
It is close to culture medium, carries out Mother culture.
2. production method according to claim 1, which is characterized in that the sawdust medium includes following mass percentage
Component: 65~75% sawdusts, 10~15% wheat bran, 8~15% maize flours, 2~5% soy noodles, 0.8~1.2% gypsum,
0.8~1.5% sugar, 0.1~0.3% potassium dihydrogen phosphate and 0.1~0.3% magnesium sulfate.
3. production method according to claim 2, which is characterized in that the sawdust medium includes following mass percentage
Component: 70% sawdust, 14.8% wheat bran, 10% maize flour, 3% soy noodle, 1% gypsum, 1% sugar, 0.1% potassium dihydrogen phosphate
With 0.1% magnesium sulfate.
4. production method described in any one according to claim 1~3, which is characterized in that the granularity of the sawdust is 5~50
Mesh.
5. production method described in any one according to claim 1~3, which is characterized in that the sterilizing side of the sawdust medium
Method high pressure steam sterilization 30min at 0.11~0.13Mpa.
6. production method according to claim 1, which is characterized in that the broadleaf deciduous tree species include Betulaceae, Salicaceae,
One of Fagaceae, Moraceae and Juglandaceae tree species are a variety of.
7. production method according to claim 1, which is characterized in that the Dictyophora rubrovalvata fructification selection 7~8 is mature, strong
The fructification of health.
8. according to claim 1 or 7 production methods, which is characterized in that the disinfection of Dictyophora rubrovalvata fructification is Gao Meng with solution
The concentration of sour potassium solution or sodium dichloro cyanurate solution, the liquor potassic permanganate or sodium dichloro cyanurate solution is 0.05
~0.1%.
9. production method according to claim 1, which is characterized in that the temperature of the Mother culture is 20~25 DEG C.
10. according to claim 1 or 9 production methods, which is characterized in that the time of the Mother culture is 25~30d.
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| CN115152531A (en) * | 2022-08-11 | 2022-10-11 | 贵州美味鲜竹荪产业有限公司 | Production method of Dictyophora rubrovalvata glass bottle mother seeds |
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Application publication date: 20190809 |