CN109937792A - A kind of artificial cultivation method of wild column ring destroying angel - Google Patents
A kind of artificial cultivation method of wild column ring destroying angel Download PDFInfo
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- CN109937792A CN109937792A CN201910219361.8A CN201910219361A CN109937792A CN 109937792 A CN109937792 A CN 109937792A CN 201910219361 A CN201910219361 A CN 201910219361A CN 109937792 A CN109937792 A CN 109937792A
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Abstract
The invention discloses a kind of artificial cultivation methods of wild column ring destroying angel.The artificial cultivation method is the following steps are included: carry out tissue separation for wild fresh column ring destroying angel, and then culture obtains inclined-plane parent species in inclined-plane PDA culture medium；Inclined-plane parent species continue culture rejuvenation in inclined-plane comprehensive PDA culture medium and obtain production parent species；Production is inoculated into culture in pedigree seed culture medium with parent species and obtains production original seed；Production is inoculated into culture in production kind culture medium with original seed and obtains production kind；Production kind is inoculated into production cultivation base, is cultivated to the fructification for growing maturation.Artificial cultivation method of the invention can make the biologicak efficiency of wild column ring destroying angel reach 35~60%, i.e., every 100g production compost can obtain the fresh column ring destroying angel of 35~60g, which can make raw material or make research material use.
Description
Technical field
The invention belongs to fungi Cultivating techniques fields, are related to a kind of artificial cultivation method of ring destroying angel, specifically
It is related to a kind of artificial cultivation method of wild column ring destroying angel.
Background technique
Ring umbrella category (Cyclocybe) is under the jurisdiction of mycota Fungi, Basidiomycota Basidiomycota, agaric in classification
Guiding principle Agaricomycetes, Agaricales Agaricales, Strophariaceae Strophariaceae, only 10 kinds of Known Species so far,
At least 5 kinds of China.The Species distributing of ring umbrella is wider, and part is all kinds of to have edible medicinal value, such as black poplar ring umbrella Cyclocybe
Aegerita (V.Brig.) Vizzini (poplar ricetulus rodents Agrocybe aegerita (V.Brig.) Singer), column ring umbrella
Cyclocybe cylindracea (DC.) Vizzini&Angelini (Agroeybe cylindracea mycelia Agrocybe cylindracea
(DC.) Maire), dark color ring umbrella Cyclocybe erebia (Fr.) Vizzini&Matheny (wet viscous ricetulus rodents Agrocybe
Erebia (Fr.) K ü hner ex Singer) He Liusheng ring umbrella Cyclocybe salicaceicola (Zhu L.Yang,
M.Zang&X.X.Liu) Vizzini (Liu Sheng ricetulus rodents (willow mushroom) Agrocybe salicaceicola Zhu L.Yang,
M.Zang&X.X.Liu[as'salicacola']).Column ring umbrella of the invention is in local civil mostly as liver protection or antitumor
And other effects eaten, wild resource is extremely limited, and so far again not yet successful artificial cultivation method.
Summary of the invention
The purpose of the present invention is to provide a kind of artificial cultivation methods of wild column ring destroying angel.
The present invention is by carrying out tissue separation for wild fresh column ring destroying angel, then in inclined-plane PDA culture medium
Upper culture obtains inclined-plane parent species；Inclined-plane parent species continue culture rejuvenation in inclined-plane comprehensive PDA culture medium and obtain production parent species；It will
Production is inoculated into culture in pedigree seed culture medium with parent species and obtains production original seed；Production is inoculated into production kind culture medium with original seed
Middle culture obtains production kind；Production kind is inoculated into production cultivation base, is cultivated to the fructification for growing maturation.
Technical solution of the present invention is as follows:
A kind of artificial cultivation method of wild column ring destroying angel, comprising the following steps:
(1) wild fresh column ring destroying angel is subjected to tissue separation, is then inoculated with by the inoculum concentration of 3~6%m/m
Into inclined-plane PDA culture medium, it is protected from light 10~18d of culture in 23~25 DEG C, covers with inclined-plane to mycelia, picking growing way is vigorous, sturdy
Uniform inclined-plane parent species or freezing are spare；The inclined-plane PDA culture medium includes the raw material of following parts by weight meter: potato
75~85 parts, 3~6 parts of sucrose, 15~25 parts of agar, 0.2~0.3 part of potassium dihydrogen phosphate, 0.1~0.2 part of magnesium sulfate, vitamin
B10.0004~0.0006 part, appropriate amount of water；
(2) the inclined-plane parent species that step (1) obtains are seeded to inclined-plane comprehensive PDA culture medium by the inoculum concentration of 3~6%m/m
In in 23~25 DEG C be protected from light 6~15d of culture, until mycelia covers with inclined-plane, choose that growing way is vigorous, sturdy uniform strain is as life
It produces and uses parent species, the inclined-plane comprehensive PDA culture medium includes the raw material of following parts by weight meter: 75~85 parts of potato, sucrose 6~
10 parts, 10~15 parts of agar, 0.2~1 part of potassium dihydrogen phosphate, 0.2~1 part of magnesium sulfate and vitamin B10.0004~0.0006
Part, appropriate amount of water；
(3) production that step (2) obtains is seeded in pedigree seed culture medium with parent species by the inoculum concentration of 3~10%m/m, in
23~25 DEG C are protected from light 12~40d of culture, until mycelia covers with pedigree seed culture medium, choose that growing way is vigorous, sturdy uniform strain is made
For production original seed, the pedigree seed culture medium is made of original seed base-material and water 1:1.0~1.3 in mass ratio, the original seed
Base-material includes the component of following weight percent meter: wheat 30~50%, sawdust 20~40%, wheat bran or rice bran 20~30%,
Sucrose 1~5% and calcium carbonate 1~3%；
(4) production that step (3) obtains is seeded in production kind culture medium with original seed by the inoculum concentration of 3~10%m/m,
In 23~25 DEG C, relative humidity 50~75% is protected from light 12~45d of culture, until mycelia covers with production kind culture medium, is cultivated
The production kind of column ring destroying angel, the production kind culture medium is by cultivation base stock and water 1:1.0~1.3 group in mass ratio
At the cultivation base stock includes the component of following weight percent meter: wheat 20~30%, sawdust 40~50%, wheat bran or
Rice bran 20~30%, sucrose 1~5% and calcium carbonate 1~3%；
(5) production kind that step (4) obtains is seeded in production cultivation base by the inoculum concentration of 5~15%m/m, Yu Wen
23~28 DEG C of degree in the cultivation room of relative humidity 50~70%, is protected from light 23~45d of culture, until mycelia covers with production cultivation
Base moves into the fructification cultivated in cultivation house to maturation is grown, the production cultivation base after then carrying out low temperature stimulation again
It is made of cultivation base stock and water 1:1.0~1.3 in mass ratio, the cultivation base stock includes the group of following weight percent meter
Point: wheat 20~30%, sawdust 40~50%, wheat bran or rice bran 20~30%, sucrose 1~5% and calcium carbonate 1~3%.
The preparation steps of the inclined-plane comprehensive PDA culture medium of the inclined-plane PDA culture medium and step (2) of the step (1) are equal are as follows:
Potato is cut into 1cm size box to be put into pot, the water of 3.5~4.5 times of its weight is added, boils 15~20min to Ma Ling
Potato it is soft without it is rotten when, with 6~8 layers of filtered through gauze, take filtered juice in pot, water added to mend to raw water amount, be added agar fusing, then plus
Enter sucrose, potassium dihydrogen phosphate, magnesium sulfate and vitamin B1It stirs evenly, is loaded on test tube while hot, after sterilizing, is cooled to 45~50 DEG C
It is spare to be put into inclined-plane.
The preparation steps of the pedigree seed culture medium of the step (3) are as follows: mixed after cultivation base stock each component is weighed respectively with water
It closes, stirs evenly, conventional sterilant of bottling is cooling spare；The production original seed general room temperature storage period, is no more than 15d, low
Gentle to put no more than 20d, expired strain cannot use.
It is preferred that the inoculum concentration of fresh column ring destroying angel wild in the step (1) is 5%m/m, when being protected from light culture
Between be 10~15d.
It is preferred that the inoculum concentration of inclined-plane parent species is 5%m/m in the step (2), being protected from light incubation time is 10~15d.
It is preferred that the inoculum concentration of production parent species is 6%m/m in the step (3), being protected from light incubation time is 15~20d.
It is preferred that the inoculum concentration of production original seed is 8%m/m in the step (4), being protected from light incubation time is 35~40d.
The preparation steps of production kind culture medium in the step (4) are as follows: after cultivation base stock each component is weighed respectively with water
Mixing, stirs evenly, conventional sterilant of bottling, and cooling is spare.
It is preferred that the inoculum concentration of production kind is 10%m/m in the step (5).
It is preferred that the production in the step (5) is made of with cultivation base cultivation base stock and water 1:1.2 in mass ratio.
The production preparation steps of cultivation base in the step (5) are as follows: after cultivation base stock each component is weighed respectively with water
Mixing, stirs evenly, and is packed into high density polyethylene (HDPE) bag or Polypropylene Bag, conventional 1.5kg/cm2Pressure sterilizes for 60 minutes, cooling
It is spare.
It is preferred that the cultivation base stock in the step (4), (5) includes the component of following weight percent meter: wheat 20%,
Sawdust 48%, wheat bran or rice bran 30%, sucrose 1% and calcium carbonate 1%.
The fructification cultivated in cultivation house to maturation is grown is moved into again after carrying out low temperature stimulation in the step (5), specifically
Are as follows: the production cultivation base for covering with mycelia is stimulated into 3~6d of culture at -5~5 DEG C, then moves into 15~20 DEG C of temperature, relatively
Humidity 65~85%, 300~600Lux of intensity of illumination aoxidize 15~20d of cultivation under the conditions of 2000~2300ppm of concentration of carbon, then
Indoor illumination intensity is moved into naturally, 15~20 DEG C of temperature, relative humidity 65~80%, the cultivation through 15~20d can grow son
Entity former base, is further cultured for 10~15d, and to fructification cap parachute-opening, collarium is not fallen off, and when spore does not spray, that is, obtains maturation
Column ring destroying angel.
Compared with prior art, the invention has the following beneficial effects:
Artificial cultivation method of the invention can make the biologicak efficiency of wild column ring destroying angel reach 35~60%, i.e.,
Every 100g production compost can obtain the fresh column ring destroying angel of 35~60g, the column ring destroying angel can make raw material or
Make research material use.
Specific embodiment
The following examples are further illustrations of the invention, rather than limiting the invention.
Embodiment 1
(1) it weighs respectively and has removed the peel fresh potato 200g, sucrose 8g, agar 40g, potassium dihydrogen phosphate 0.53g, sulfuric acid
Magnesium 0.27g and vitamin B10.001g；The square that potato is cut into 1cm size is put into pot, 870g water is added, boils
15min to potato it is soft without it is rotten when, with 6 layers of filtered through gauze, take filtered juice in pot, water added to mend to raw water amount, be added agar
Fusing, then plus sucrose, potassium dihydrogen phosphate, magnesium sulfate and vitamin B1It stirs evenly, after sterilizing while hot loaded on test tube, is cooled to 45
DEG C it is put into inclined-plane, obtains inclined-plane PDA culture medium.
(2) it weighs respectively and has removed the peel fresh potato 200g, sucrose 16g, agar 26.7g, potassium dihydrogen phosphate 0.53g,
Magnesium sulfate 0.53g and vitamin B1 0.001g, the square that potato is cut into 1cm size is put into pot, and 853g water is added, boils
Boil 15min to potato it is soft without it is rotten when, with 6 layers of filtered through gauze, take filtered juice in pot, add water to mend to raw water amount, add agar
Fusing, then plus sucrose, potassium dihydrogen phosphate, magnesium sulfate and vitamin B1It stirs evenly, after sterilizing while hot loaded on test tube, is cooled to 45
DEG C it is put into inclined-plane, obtains inclined-plane comprehensive PDA culture medium.
(3) wheat 300g, sawdust 400g, wheat bran 220g are weighed respectively, and sucrose 50g, calcium carbonate 30g are mixed to get 1000g
Original seed base-material is added 1200g water, stirs evenly, and conventional sterilant of bottling, cooling is spare, obtains pedigree seed culture medium.
(4) wheat 3000g, sawdust 4000g, wheat bran 2200g are weighed respectively, and sucrose 500g, calcium carbonate 300g are mixed to get
10000g cultivation base stock is added 11000g water, stirs evenly, and conventional sterilant of bottling, cooling is spare, obtains production kind culture medium.
(5) wheat 30000g, sawdust 40000g, wheat bran 22000g, sucrose 5000g, calcium carbonate 3000g are weighed respectively, are mixed
Conjunction obtains 100kg cultivation base stock, and 110kg water is added, stirs evenly, and is packed into high density polyethylene (HDPE) bag, conventional 1.5kg/cm2Pressure
Power, 60min sterilizing, cooling is spare, obtains production cultivation base.
(6) wild fresh column ring destroying angel is subjected to tissue separation, about 15g is taken to be inoculated in the inclined-plane 300g PDA training
It supports in base, culture 10d is protected from light in 23 DEG C, cover with inclined-plane to mycelia, picking growing way is vigorous, sturdy uniform inclined-plane parent species or freezing
Preservation is spare.
(7) the inclined-plane parent species of 50g column ring umbrella are seeded in the inclined-plane 1000g comprehensive PDA culture medium, are protected from light training in 23 DEG C
12d is supported, until mycelia covers with inclined-plane, chooses the production parent species that growing way is vigorous, sturdy uniform strain is as column ring umbrella.
(8) production of 132g column ring umbrella is seeded in 2200g pedigree seed culture medium with parent species, is protected from light culture in 23 DEG C
15d, until mycelia covers with pedigree seed culture medium, selection growing way is vigorous, sturdy uniform strain is former as the production of column ring umbrella
Kind.
(9) production of 1920g column ring umbrella is seeded in 24kg production kind culture medium with original seed, it is relatively wet in 23 DEG C
Degree 50% is protected from light culture 12d, until mycelia covers with production kind culture medium, obtains the production kind of column ring umbrella.
(10) production kind of 22kg column ring umbrella is inoculated in 220kg production cultivation base, 23 DEG C of Yu Wendu, relative humidity
In 50% cultivation room, it is protected from light culture 35d, until mycelia covers with production cultivation base；The production cultivation base of mycelia will be covered with
3d are cultivated in -5~5 DEG C of stimulations, then move into 15~20 DEG C of temperature, relative humidity 65~85%, intensity of illumination 300~
600Lux, oxidation concentration of carbon cultivate 15d under the conditions of 2000~2300ppm, then move into indoor illumination intensity naturally, temperature 15~
20 DEG C, relative humidity 65~80%, the cultivation through 15d can grow fruit body primordium, be further cultured for 12d, open to fructification cap
Umbrella, collarium are not fallen off, and when spore does not spray, can harvest to obtain column ring destroying angel, at this time with sharp cutter by fructification
It is cut off along stem base portion, biological transformation ratio can reach 35%, and the fructification under adopting will be handled in time, or dry under sunlight
Or drying and dehydrating and packing.
Embodiment 2
(1) it weighs respectively and has removed the peel fresh potato 1275g, sucrose 90g, agar 375g, potassium dihydrogen phosphate 4.5g, sulphur
Sour magnesium 3g and vitamin B10.009g；The square that potato is cut into 1cm size is put into pot, 7864g water is added, boils
20min to potato it is soft without it is rotten when, with 8 layers of filtered through gauze, take filtered juice in pot, water added to mend to raw water amount, be added agar
Fusing, then plus sucrose, potassium dihydrogen phosphate, magnesium sulfate and vitamin B1It stirs evenly, after sterilizing while hot loaded on test tube, is cooled to 50
DEG C it is put into inclined-plane, obtains inclined-plane PDA culture medium.
(2) it weighs respectively and has removed the peel fresh potato 1275g, sucrose 150g, agar 225g, potassium dihydrogen phosphate 15g, sulphur
Sour magnesium 15g and vitamin B1 0.009g, the square that potato is cut into 1cm size is put into pot, and 7560g water is added, boils
20min to potato it is soft without it is rotten when, with 8 layers of filtered through gauze, take filtered juice in pot, water added to mend to raw water amount, add agar molten
Change, then plus sucrose, potassium dihydrogen phosphate, magnesium sulfate and vitamin B1It stirs evenly, after sterilizing while hot loaded on test tube, is cooled to 50 DEG C
It is put into inclined-plane, obtains inclined-plane comprehensive PDA culture medium.
(3) wheat 375g, sawdust 210g, wheat bran 150g are weighed respectively, and sucrose 7.5g, calcium carbonate 7.5g are mixed to get
750g original seed base-material is added 825g water, stirs evenly, and conventional sterilant of bottling, cooling is spare, obtains pedigree seed culture medium.
(4) wheat 2100g, sawdust 3750g, wheat bran 1500g are weighed respectively, and sucrose 75g, calcium carbonate 75g are mixed to get
7500g cultivation base stock is added 9000g water, stirs evenly, and conventional sterilant of bottling, cooling is spare, obtains production kind culture medium.
(5) wheat 8400g, sawdust 15000g, wheat bran 6000g are weighed respectively, and sucrose 300g, calcium carbonate 300g are mixed
To 30kg cultivation base stock, 39kg water is added, stirs evenly, is packed into high density polyethylene (HDPE) bag, conventional 1.5kg/cm2Pressure, 60min
Sterilizing, cooling is spare, obtains production cultivation base.
(6) wild fresh column ring destroying angel is subjected to tissue separation, about 15g is taken to be inoculated in the inclined-plane 300g PDA training
It supports in base, culture 15d is protected from light in 25 DEG C, cover with inclined-plane to mycelia, picking growing way is vigorous, sturdy uniform inclined-plane parent species or freezing
Preservation is spare.
(7) the inclined-plane parent species of 24g column ring umbrella are seeded in the inclined-plane 600g comprehensive PDA culture medium, are protected from light training in 23 DEG C
15d is supported, until mycelia covers with inclined-plane, chooses the production parent species that growing way is vigorous, sturdy uniform strain is as column ring umbrella.
(8) production of 150g column ring umbrella is seeded in 1500g pedigree seed culture medium with parent species, is protected from light culture in 25 DEG C
40d, until mycelia covers with pedigree seed culture medium, selection growing way is vigorous, sturdy uniform strain is former as the production of column ring umbrella
Kind.
(9) production of 1500g column ring umbrella is seeded in 15000g production kind culture medium with original seed, in 25 DEG C, relatively
Humidity 60% is protected from light culture 45d, until mycelia covers with production kind culture medium, obtains the production kind of column ring umbrella.
(10) production kind of 10800g column ring umbrella is inoculated in 72000g production cultivation base, 23 DEG C of Yu Wendu, relatively
In the cultivation room of humidity 60%, it is protected from light culture 45d, until mycelia covers with production cultivation base；The production for covering with mycelia is planted
Pei Ji cultivates 6d in -5~5 DEG C of stimulations, then moves into 15~20 DEG C of temperature, relative humidity 65~85%, and intensity of illumination 300~
600Lux, oxidation concentration of carbon cultivate 20d under the conditions of 2000~2300ppm, then move into indoor illumination intensity naturally, temperature 15~
20 DEG C, relative humidity 65~80%, the cultivation through 20d can grow fruit body primordium, be further cultured for 10d, open to fructification cap
Umbrella, collarium are not fallen off, and when spore does not spray, can harvest to obtain column ring destroying angel, at this time with scissors by fructification along bacterium
It is cut at handle base portion, biological transformation ratio can reach 48%, and the fructification under adopting will be handled in time, or dries or do under sunlight
Dry dehydration and packing.
Embodiment 3
(1) it weighs respectively and has removed the peel fresh potato 1600g, sucrose 100g, agar 400g, potassium dihydrogen phosphate 4g, sulphur
Sour magnesium 4g and vitamin B10.01g；The square that potato is cut into 1cm size is put into pot, 8432g water is added, boils
15min to potato it is soft without it is rotten when, with 6 layers of filtered through gauze, take filtered juice in pot, water added to mend to raw water amount, be added agar
Fusing, then plus sucrose, potassium dihydrogen phosphate, magnesium sulfate and vitamin B1It stirs evenly, after sterilizing while hot loaded on test tube, is cooled to 45
DEG C it is put into inclined-plane, obtains inclined-plane PDA culture medium.
(2) it weighs respectively and has removed the peel fresh potato 1600g, sucrose 160g, agar 240g, potassium dihydrogen phosphate 10g, sulphur
Sour magnesium 10g and vitamin B1 0.01g, the square that potato is cut into 1cm size is put into pot, and 8080g water is added, boils
15min to potato it is soft without it is rotten when, with 6 layers of filtered through gauze, take filtered juice in pot, water added to mend to raw water amount, add agar molten
Change, then plus sucrose, potassium dihydrogen phosphate, magnesium sulfate and vitamin B1It stirs evenly, after sterilizing while hot loaded on test tube, is cooled to 45 DEG C
It is put into inclined-plane, obtains inclined-plane comprehensive PDA culture medium.
(3) wheat 480g, sawdust 200g, wheat bran 300g are weighed respectively, and sucrose 10g, calcium carbonate 10g are mixed to get 1000g
Original seed base-material is added 1300g water, stirs evenly, and conventional sterilant of bottling, cooling is spare, obtains pedigree seed culture medium.
(4) wheat wheat 2000g, sawdust 4800g, rice bran 3000g, sucrose 100g, calcium carbonate 100g mixing are weighed respectively
10000g cultivation base stock is obtained, 13000g water is added, stirs evenly, conventional sterilant of bottling, cooling is spare, obtains production kind culture
Base.
(5) wheat 8000g, sawdust 19200g, rice bran 12000g are weighed respectively, and sucrose 400g, calcium carbonate 400g are mixed
To 40000g cultivation base stock, 52000kg water is added, stirs evenly, is packed into high density polyethylene (HDPE) bag, conventional 1.5kg/cm2Pressure,
60min sterilizing, cooling is spare, obtains production cultivation base.
(6) wild fresh column ring destroying angel is subjected to tissue separation, 18g is taken to be inoculated in the inclined-plane 300g PDA culture
In base, it is protected from light culture 14d in 23 DEG C, covers with inclined-plane to mycelia, picking growing way is vigorous, sturdy uniform inclined-plane parent species or freezing are protected
It hides spare.
(7) the inclined-plane parent species of 96g column ring umbrella are seeded in the inclined-plane 1600g comprehensive PDA culture medium, are protected from light training in 25 DEG C
15d is supported, until mycelia covers with inclined-plane, chooses the production parent species that growing way is vigorous, sturdy uniform strain is as column ring umbrella.
(8) production of 100g column ring umbrella is seeded in 2000g pedigree seed culture medium with parent species, is protected from light culture in 25 DEG C
12d, until mycelia covers with pedigree seed culture medium, selection growing way is vigorous, sturdy uniform strain is former as the production of column ring umbrella
Kind.
(9) production of 1800g column ring umbrella is seeded in 18000g production kind culture medium with original seed, in 23 DEG C, relatively
Humidity 60% is protected from light culture 23d, until mycelia covers with production kind culture medium, obtains the production kind of column ring umbrella.
(10) production kind of 9000g column ring umbrella is inoculated in 90000g production cultivation base, it is 25 DEG C of Yu Wendu, relatively wet
In the cultivation room of degree 60%, it is protected from light culture 23d, until mycelia covers with production cultivation base；The production cultivation of mycelia will be covered with
Base cultivates 5d in -5~5 DEG C of stimulations, then moves into 15~20 DEG C of temperature, relative humidity 65~85%, and intensity of illumination 300~
600Lux, oxidation concentration of carbon cultivate 18d under the conditions of 2000~2300ppm, then move into indoor illumination intensity naturally, temperature 15~
20 DEG C, relative humidity 65~80%, the cultivation through 18d can grow fruit body primordium, be further cultured for 15d, open to fructification cap
Umbrella, collarium are not fallen off, and when spore does not spray, can harvest to obtain column ring destroying angel, at this time with sharp cutter by fructification
It is cut off along stem base portion, biological transformation ratio can reach 60%, and the fructification under adopting will be handled in time, or dry under sunlight
Or drying and dehydrating and packing.
Embodiment 4
(1) it weighs respectively and has removed the peel fresh potato 200g, sucrose 8g, agar 40g, potassium dihydrogen phosphate 0.53g, sulfuric acid
Magnesium 0.27g and vitamin B10.001g；The square that potato is cut into 1cm size is put into pot, 870g water is added, boils
15min to potato it is soft without it is rotten when, with 6 layers of filtered through gauze, take filtered juice in pot, water added to mend to raw water amount, be added agar
Fusing, then plus sucrose, potassium dihydrogen phosphate, magnesium sulfate and vitamin B1It stirs evenly, after sterilizing while hot loaded on test tube, is cooled to 45
DEG C it is put into inclined-plane, obtains inclined-plane PDA culture medium.
(2) it weighs respectively and has removed the peel fresh potato 200g, sucrose 16g, agar 26.7g, potassium dihydrogen phosphate 0.53g,
Magnesium sulfate 0.53g and vitamin B1 0.001g, the square that potato is cut into 1cm size is put into pot, and 853g water is added, boils
Boil 15min to potato it is soft without it is rotten when, with 6 layers of filtered through gauze, take filtered juice in pot, add water to mend to raw water amount, add agar
Fusing, then plus sucrose, potassium dihydrogen phosphate, magnesium sulfate and vitamin B1It stirs evenly, after sterilizing while hot loaded on test tube, is cooled to 45
DEG C it is put into inclined-plane, obtains inclined-plane comprehensive PDA culture medium.
(3) wheat 900g, sawdust 600g, rice bran 400g are weighed respectively, and sucrose 60g, calcium carbonate 40g are mixed to get 2000g
Original seed base-material is added 2400g water, stirs evenly, and conventional sterilant of bottling, cooling is spare, obtains pedigree seed culture medium.
(4) wheat 6000g, sawdust 9000g, rice bran 4000g are weighed respectively, and sucrose 600g, calcium carbonate 400g are mixed to get
30000g cultivation base stock is added 36000g water, stirs evenly, and conventional sterilant of bottling, cooling is spare, obtains production kind culture medium.
(5) wheat 24000g, sawdust 36000g, rice bran 16000g, sucrose 2400g, calcium carbonate 1600g mixing are weighed respectively
80000g cultivation base stock is obtained, 104000g water is added, stirs evenly, is packed into high density polyethylene (HDPE) bag, conventional 1.5kg/cm2Pressure
Power, 60min sterilizing, cooling is spare, obtains production cultivation base.
(6) wild fresh column ring destroying angel is subjected to tissue separation, 36g is taken to be inoculated in the inclined-plane 1200g PDA culture
In base, it is protected from light culture 18d in 25 DEG C, covers with inclined-plane to mycelia, picking growing way is vigorous, sturdy uniform inclined-plane parent species or freezing are protected
It hides spare.
(7) the inclined-plane parent species of 240g column ring umbrella are seeded in the inclined-plane 8000g comprehensive PDA culture medium, are protected from light in 23 DEG C
6d is cultivated, until mycelia covers with inclined-plane, chooses the production parent species that growing way is vigorous, sturdy uniform strain is as column ring umbrella.
(8) production of 132g column ring umbrella is seeded in 4400g pedigree seed culture medium with parent species, is protected from light culture in 25 DEG C
18d, until mycelia covers with pedigree seed culture medium, selection growing way is vigorous, sturdy uniform strain is former as the production of column ring umbrella
Kind.
(9) production of 1200g column ring umbrella is seeded in 40000g production kind culture medium with original seed, in 25 DEG C, relatively
Humidity 75% is protected from light culture 25d, until mycelia covers with production kind culture medium, obtains the production kind of column ring umbrella.
(10) production kind of 9000g column ring umbrella is inoculated in 180000g production cultivation base, 23 DEG C of Yu Wendu, relatively
In the cultivation room of humidity 70%, it is protected from light culture 25d, until mycelia covers with production cultivation base；The production for covering with mycelia is planted
Pei Ji cultivates 4d in -5~5 DEG C of stimulations, then moves into 15~20 DEG C of temperature, relative humidity 65~85%, and intensity of illumination 300~
600Lux, oxidation concentration of carbon cultivate 15d under the conditions of 2000~2300ppm, then move into indoor illumination intensity naturally, temperature 15~
20 DEG C, relative humidity 65~80%, the cultivation through 20d can grow fruit body primordium, be further cultured for 15d, open to fructification cap
Umbrella, collarium are not fallen off, and when spore does not spray, can harvest to obtain column ring destroying angel, at this time with sharp cutter by fructification
It is cut off along stem base portion, biological transformation ratio can reach 53%, and the fructification under adopting will be handled in time, or dry under sunlight
Or drying and dehydrating and packing.
The above is only the preferred embodiment of the present invention, it is noted that above-mentioned preferred embodiment is not construed as pair
Limitation of the invention, protection scope of the present invention should be defined by the scope defined by the claims..For the art
For those of ordinary skill, without departing from the spirit and scope of the present invention, several improvements and modifications can also be made, these change
It also should be regarded as protection scope of the present invention into retouching.
Claims (10)
1. a kind of artificial cultivation method of wild column ring destroying angel, which comprises the following steps:
Wild fresh column ring destroying angel is subjected to tissue separation, then culture obtains inclined-plane in inclined-plane PDA culture medium
Parent species；Inclined-plane parent species continue culture rejuvenation in inclined-plane comprehensive PDA culture medium and obtain production parent species；Production is inoculated with parent species
Into pedigree seed culture medium, culture obtains production original seed；Production is inoculated into cultivate in production kind culture medium with original seed and is produced
Kind；Production kind is inoculated into production cultivation base, is cultivated to the fructification for growing maturation.
2. the artificial cultivation method of wild column ring destroying angel according to claim 1, which is characterized in that including following
Step:
(1) wild fresh column ring destroying angel is subjected to tissue separation, is then inoculated into tiltedly by the inoculum concentration of 3~6%m/m
In the PDA culture medium of face, it is protected from light 10~18d of culture in 23~25 DEG C, covers with inclined-plane to mycelia, picking growing way is vigorous, sturdy uniform
Inclined-plane parent species or freezing it is spare；The inclined-plane PDA culture medium includes the raw material of following parts by weight meter: potato 75~
85 parts, 3~6 parts of sucrose, 15~25 parts of agar, 0.2~0.3 part of potassium dihydrogen phosphate, 0.1~0.2 part of magnesium sulfate, vitamin B1
0.0004~0.0006 part, appropriate amount of water；
(2) by inclined-plane parent species that step (1) obtains by the inoculum concentration of 3~6%m/m be seeded in inclined-plane comprehensive PDA culture medium in
23~25 DEG C are protected from light 6~15d of culture, until mycelia covers with inclined-plane, choose that growing way is vigorous, sturdy uniform strain is used as production
Parent species, the inclined-plane comprehensive PDA culture medium include the raw material of following parts by weight meter: 75~85 parts of potato, sucrose 6~10
Part, 10~15 parts of agar, 0.2~1 part of potassium dihydrogen phosphate, 0.2~1 part of magnesium sulfate and vitamin B10.0004~0.0006 part,
Appropriate amount of water；
(3) production that step (2) obtains is seeded in pedigree seed culture medium with parent species by the inoculum concentration of 3~10%m/m, in 23~
25 DEG C are protected from light 12~40d of culture, until mycelia covers with pedigree seed culture medium, choose that growing way is vigorous, sturdy uniform strain is as life
It produces and uses original seed, the pedigree seed culture medium is made of original seed base-material and water 1:1.0~1.3 in mass ratio, the original seed base-material
Component including following weight percent meter: wheat 30~50%, sawdust 20~40%, wheat bran or rice bran 20~30%, sucrose 1
~5% and calcium carbonate 1~3%；
(4) production that step (3) obtains is seeded in production kind culture medium with original seed by the inoculum concentration of 3~10%m/m, in 23
~25 DEG C, relative humidity 50~75% is protected from light 12~45d of culture, until mycelia covers with production kind culture medium, obtains cultivation column
The production kind of ring destroying angel, the production kind culture medium are made of cultivation base stock and water 1:1.0~1.3 in mass ratio,
The cultivation base stock includes the component of following weight percent meter: wheat 20~30%, sawdust 40~50%, wheat bran or rice bran
20~30%, sucrose 1~5% and calcium carbonate 1~3%；
(5) production kind that step (4) obtains is seeded in production cultivation base by the inoculum concentration of 5~15%m/m, Yu Wendu 23
~25 DEG C, in the cultivation room of relative humidity 50~70%, it is protected from light 23~45d of culture, until mycelia covers with production cultivation base, so
The fructification cultivated in cultivation house to maturation is grown is moved into again after carrying out low temperature stimulation afterwards, and the production cultivation base is by cultivating
Base-material and water 1:1.0 in mass ratio~1.3 composition, the cultivation base stock includes the component of following weight percent meter: wheat
20~30%, sawdust 40~50%, wheat bran or rice bran 20~30%, sucrose 1~5% and calcium carbonate 1~3%.
3. the artificial cultivation method of wild column ring destroying angel according to claim 2, which is characterized in that the step
(1) preparation steps of the inclined-plane comprehensive PDA culture medium of inclined-plane PDA culture medium and step (2) are equal are as follows: potato is cut into 1cm
Size box is put into pot, and the water of 3.5~4.5 times of its weight is added, boil 15~20min to potato it is soft without it is rotten when, with 6
~8 layers of filtered through gauze, take filtered juice in pot, and water is added to mend to raw water amount, and agar fusing is added, adds sucrose, biphosphate
Potassium, magnesium sulfate and vitamin B1It stirs evenly, is loaded on test tube while hot, after sterilizing, being cooled to 45~50 DEG C, to be put into inclined-plane spare.
4. the artificial cultivation method of wild column ring destroying angel according to claim 2, which is characterized in that the step
(1) inoculum concentration of wild fresh column ring destroying angel is 5%m/m in, and being protected from light incubation time is 10~15d.
5. the artificial cultivation method of wild column ring destroying angel according to claim 2, which is characterized in that the step
(2) inoculum concentration of inclined-plane parent species is 5%m/m in, and being protected from light incubation time is 10~15d.
6. the artificial cultivation method of wild column ring destroying angel according to claim 2, which is characterized in that the step
(3) inoculum concentration of production parent species is 6%m/m in, and being protected from light incubation time is 15~20d.
7. the artificial cultivation method of wild column ring destroying angel according to claim 2, which is characterized in that the step
(4) inoculum concentration of production original seed is 8%m/m in, and being protected from light incubation time is 35~40d.
8. the artificial cultivation method of wild column ring destroying angel according to claim 2, which is characterized in that the step
(5) inoculum concentration of production kind is 10%m/m in.
9. the artificial cultivation method of wild column ring destroying angel according to claim 2, which is characterized in that the step
(4), the cultivation base stock in (5) includes the component of following weight percent meter: wheat 20%, sawdust 48%, wheat bran or rice bran
30%, sucrose 1% and calcium carbonate 1%.
10. the artificial cultivation method of wild column ring destroying angel according to claim 2, which is characterized in that the step
Suddenly the fructification cultivated in cultivation house to maturation is grown is moved into again after carrying out low temperature stimulation in (5), specifically: mycelia will be covered with
Production cultivation base stimulates 3~6d of culture at -5~5 DEG C, then moves into 15~20 DEG C of temperature, relative humidity 65~85%, light
According to 300~600Lux of intensity, 15~20d of cultivation under the conditions of 2000~2300ppm of concentration of carbon is aoxidized, then moves into indoor illumination intensity
Naturally, 15~20 DEG C of temperature, relative humidity 65~80%, the cultivation through 15~20d can grow fruit body primordium, be further cultured for 10
~15d, to fructification cap parachute-opening, collarium is not fallen off, and when spore does not spray, that is, obtains mature column ring destroying angel.
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Address after: 510070 No.56 courtyard, No.100 Xianlie Middle Road, Yuexiu District, Guangzhou City, Guangdong Province Patentee after: Institute of Microbiology, Guangdong Academy of Sciences Address before: 510070 No.56 courtyard, No.100 Xianlie Middle Road, Yuexiu District, Guangzhou City, Guangdong Province Patentee before: GUANGDONG INSTITUTE OF MICROBIOLOGY (GUANGDONG DETECTION CENTER OF MICROBIOLOGY) |