CN107360859A - A kind of artificial cultivation method of cepe - Google Patents
A kind of artificial cultivation method of cepe Download PDFInfo
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- CN107360859A CN107360859A CN201710738731.XA CN201710738731A CN107360859A CN 107360859 A CN107360859 A CN 107360859A CN 201710738731 A CN201710738731 A CN 201710738731A CN 107360859 A CN107360859 A CN 107360859A
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- cepe
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- lophatherum gracile
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/14—Fungi; Culture media therefor
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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
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- C—CHEMISTRY; METALLURGY
- C05—FERTILISERS; MANUFACTURE THEREOF
- C05B—PHOSPHATIC FERTILISERS
- C05B1/00—Superphosphates, i.e. fertilisers produced by reacting rock or bone phosphates with sulfuric or phosphoric acid in such amounts and concentrations as to yield solid products directly
- C05B1/02—Superphosphates
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Abstract
The present invention discloses a kind of artificial cultivation method of cepe, comprises the following steps:A. parent species are separately cultured;B. strain expands numerous;C. lophatherum gracile cultivating aseptic seedling;D. mycorrhiza fungi seeding cultivating and cultivation fruiting:The bacterium ball for expanding numerous acquisition is broken into bacterium slurry, dilutes 10 50 times, cultivation again is injected into after lophatherum gracile aseptic seedling until being trained Va Mycorrhiza Seedling, then Va Mycorrhiza Seedling is implanted into earthing mushroom bed, cultivate fruiting.Beneficial effects of the present invention are:The artificial cultivation of cepe is realized by the mycorhiza structure of external structure lophatherum gracile and cepe, cepe can be improved and expand the anti-adversity ability of breeding, while improve the content of each nutritional ingredient of cepe itself;Conservation of nature ecological environment and wild resource, while cultivation technique is simple to operate, cost is low, it is easy to spread, the industrialization cultivation of cepe can be realized.
Description
Technical field
The invention belongs to edible fungus technical field of cultivation, and in particular to a kind of artificial cultivation method of cepe.
Background technology
Cepe (Boletus edulis Ball.ex Fr) belongs to Basidiomycotina (Basidiom yco
Tina), one in Hymenomycetes (Hym enomy cetes), Agaricales (Aga ricases), Boletaceae (Boletaceae)
Kind, it is a kind of higher fungi of the food medicine dual-purpose economic value of distribution in world wide.Each province of China is distributed, southwest
Yield is higher.Fleshy hypertrophy and delicate, aromatic flavour, delicious flavour, the nutritious, unique flavor of cepe.Except tool
There is certain nutritive value, and there is higher medical value, zoopery and clinical study results show cepe
Containing a variety of alkaloids can control lumbocrural pain, numb in every limb, disk bone do not relax, the disease such as tic of limbs and women's abnormal leucorrhea;Its
In polysaccharide and basic protein can be antitumor, viral, adjust the immunologic function of human body.It is beautiful as a kind of precious wild mushrooms
Taste bolete be China it is important go out mushroom class, but because it is a kind of with higher plant symbiosis mycorrhizal fungi, it is difficult to
Grown on synthetic medium, the cepe of commerce and trade at present, can not all from the wild resource gathered in forest
Meets the needs of growing.
The content of the invention
To solve the above-mentioned problems in the prior art, it is an object of the invention to provide a kind of people of cepe
Work cultural method, to realize the Planting of cepe by the mycorhiza structure of external structure lophatherum gracile and cepe
Training, while improve the content of each nutritional ingredient of cepe.
For achieving the above object, the technical solution adopted by the present invention is as follows:
A kind of artificial cultivation method of cepe, comprises the following steps:
A. parent species are separately cultured:Fresh, sturdy, medium ripe wild fructification is gathered, is cleaned, surface sterilization, Yu Wu
Cutting tissue block is cultivated in mother culture media under the conditions of bacterium;
B. strain expands numerous:Step a is cultivated into the access of gained parent species containing shaken cultivation in the container for expanding breeding culture medium, oscillation frequency
Rate is 10-30rpm/min;
C. lophatherum gracile cultivating aseptic seedling:Ripe lophatherum gracile bamboo rice is selected, sterilization of shelling is placed on inducing culture after rinsing
Culture, lophatherum gracile regeneration plant i.e. lophatherum gracile aseptic seedling is obtained after callus is induced through subculture and transdifferentiation culture;
D. mycorrhiza fungi seeding cultivating and cultivation fruiting:The bacterium ball for expanding numerous acquisition is broken into bacterium slurry, 10-50 times is diluted, is injected into
Va Mycorrhiza Seedling up to being trained Va Mycorrhiza Seedling, is then implanted into earthing mushroom bed by cultivation again after lophatherum gracile aseptic seedling, cultivates fruiting.
One embodiment wherein, methods described step a tissue block is stem, cap or cap and stem intersection.
One embodiment wherein, methods described step a mother culture media is potato 160-180g, glucose 15-
17g, Yeast diffusion juice 1.6-1.8g, vitamin B126-32ug, water 900-1000ml.
One embodiment wherein, methods described step b expansion breeding culture medium is dark brown earth 4-4.5kg, wheat 4-
4.5kg, birch wood chip 1.5-2.5kg, turf 0.6-1kg, potassium sulfate 0.1-0.2kg, calcium superphosphate 0.1-0.2kg, phoenix-tail fern
Extract solution 3-7ml.
One embodiment wherein, the extracting method of the phoenix-tail fern extract solution are to crush phoenix-tail fern.Cross 40-50 mesh
Sieve, weighs powder 40-50g, adds refluxing extraction 3-5 times at 60-70 DEG C of the 70-90% ethanol of 2-4 times of volume, 1.5-2h/ times,
Merging filtrate, centrifugation, takes supernatant, and it is alcohol extract to volatilize after solvent, alcohol extract is dissolved with absolute ethyl alcohol, after distilled water diluting
PH is adjusted to neutrality, 4 DEG C save backup after sterilizing.
One embodiment wherein, the soil used in methods described step b earthing mushroom bed is scpiolitc powder, ash
Sandy soil, dark brown earth are with 1:2-3:2.5-4 ratio mix.
Tissue stem, cap or cap and stem intersection of the present invention first with the stronger cepe of growing power
Cultivate parent species, injected after expanding the numerous ball into bacterium by way of injection in lophatherum gracile aseptic seedling root, make its quickly contaminate lophatherum gracile without
Vaccine root, shorten the cultivation period of Va Mycorrhiza Seedling;On the one hand phoenix-tail fern extract solution is added in breeding culture medium is expanded in addition can strengthen
Expand the anti-adversity ability of breeding, on the other hand can supply protein, hormone, shallow lake that cepe needs to absorb in plant roots
The nutrition such as powder, alkaloid.
Compared with prior art, beneficial effects of the present invention are:Pass through external structure lophatherum gracile and the bacterium of cepe
Root architecture realizes the artificial cultivation of cepe, can improve cepe and expand the anti-adversity ability of breeding, while improve delicious food
The content of each nutritional ingredient of bolete itself;Conservation of nature ecological environment and wild resource, at the same cultivation technique it is simple to operate, into
This is low, easy to spread, can realize the industrialization cultivation of cepe.
Embodiment
To make those skilled in the art be better understood when technical scheme, with reference to embodiment pair
Technical solution of the present invention is described in detail.
Experimental method used in following embodiments is conventional method unless otherwise specified.
Material used, reagent etc., unless otherwise specified, are commercially obtained in following embodiments.
Embodiment 1
A. parent species are separately cultured:Fresh, sturdy, the medium wild fructification of ripe cepe is gathered, is cleaned, surface
Sterilization, stem tissue block is cut under aseptic condition and is cultivated in mother culture media, the mother culture media be potato 160g,
Glucose 15g, Yeast diffusion juice 1.6g, vitamin B126ug, water 900ml;
B. strain expands numerous:Step a is cultivated into the access of gained parent species containing shaken cultivation in the container for expanding breeding culture medium, oscillation frequency
Rate is 10rpm/min, and expansion breeding culture medium is dark brown earth 4kg, wheat 4kg, birch wood chip 1.5kg, turf 0.6kg, potassium sulfate
0.1kg, calcium superphosphate 0.1kg, phoenix-tail fern extract solution 3ml;
C. lophatherum gracile cultivating aseptic seedling:Ripe lophatherum gracile bamboo rice is selected, sterilization of shelling is placed on inducing culture after rinsing
Culture, lophatherum gracile regeneration plant i.e. lophatherum gracile aseptic seedling is obtained after callus is induced through subculture and transdifferentiation culture;
D. mycorrhiza fungi seeding cultivating and cultivation fruiting:The bacterium ball for expanding numerous acquisition is broken into bacterium slurry, 10 times is diluted, is injected into henon bamboo
Va Mycorrhiza Seedling up to being trained Va Mycorrhiza Seedling, is then implanted into earthing mushroom bed by cultivation again after leaf aseptic seedling, fruiting is cultivated, wherein covering
The soil used in native mushroom bed is scpiolitc powder, grey sandy soil, dark brown earth with 1:2:2.5 ratio mixes.
Embodiment 2
A. parent species are separately cultured:Fresh, sturdy, medium ripe wild fructification is gathered, is cleaned, surface sterilization, Yu Wu
Cut cap tissue block under the conditions of bacterium to cultivate in mother culture media, the mother culture media is potato 170g, glucose
16g, Yeast diffusion juice 1.7g, vitamin B129ug, water 950ml;
B. strain expands numerous:Step a is cultivated into the access of gained parent species containing shaken cultivation in the container for expanding breeding culture medium, oscillation frequency
Rate is 20rpm/min, and expansion breeding culture medium is dark brown earth 4.25kg, wheat 4.25kg, birch wood chip 2kg, turf 0.8kg, sulfuric acid
Potassium 0.15kg, calcium superphosphate 0.15kg, phoenix-tail fern extract solution 5ml;
C. lophatherum gracile cultivating aseptic seedling:Ripe lophatherum gracile bamboo rice is selected, sterilization of shelling is placed on inducing culture after rinsing
Culture, lophatherum gracile regeneration plant i.e. lophatherum gracile aseptic seedling is obtained after callus is induced through subculture and transdifferentiation culture;
D. mycorrhiza fungi seeding cultivating and cultivation fruiting:The bacterium ball for expanding numerous acquisition is broken into bacterium slurry, 30 times is diluted, is injected into henon bamboo
Va Mycorrhiza Seedling up to being trained Va Mycorrhiza Seedling, is then implanted into earthing mushroom bed by cultivation again after leaf aseptic seedling, fruiting is cultivated, wherein covering
The soil used in native mushroom bed is scpiolitc powder, grey sandy soil, dark brown earth with 1:2.5:3.25 ratio mixes.
Embodiment 3
A. parent species are separately cultured:Fresh, sturdy, medium ripe wild fructification is gathered, is cleaned, surface sterilization, Yu Wu
Stem, cap or cap are cut under the conditions of bacterium and stem intersection tissue block is cultivated in mother culture media, the Mother culture
Base is potato 180g, glucose 17g, Yeast diffusion juice 1.8g, vitamin B132ug, water 1000ml;
B. strain expands numerous:Step a is cultivated into the access of gained parent species containing shaken cultivation in the container for expanding breeding culture medium, oscillation frequency
Rate is 30rpm/min, and expansion breeding culture medium is dark brown earth 4.5kg, wheat 4.5kg, birch wood chip 2.5kg, turf 1kg, potassium sulfate
0.2kg, calcium superphosphate 0.2kg, phoenix-tail fern extract solution 7ml;
C. lophatherum gracile cultivating aseptic seedling:Ripe lophatherum gracile bamboo rice is selected, sterilization of shelling is placed on inducing culture after rinsing
Culture, lophatherum gracile regeneration plant i.e. lophatherum gracile aseptic seedling is obtained after callus is induced through subculture and transdifferentiation culture;
D. mycorrhiza fungi seeding cultivating and cultivation fruiting:The bacterium ball for expanding numerous acquisition is broken into bacterium slurry, 50 times is diluted, is injected into henon bamboo
Va Mycorrhiza Seedling up to being trained Va Mycorrhiza Seedling, is then implanted into earthing mushroom bed by cultivation again after leaf aseptic seedling, fruiting is cultivated, wherein covering
The soil used in native mushroom bed is scpiolitc powder, grey sandy soil, dark brown earth with 1:3:4 ratio mixes.
Comparative example 1
In addition to phoenix-tail fern extract solution is not added in step b expansion breeding culture medium, other cultivation conditions are the same as embodiment 2.
Phoenix-tail fern extract solution can be prepared by the following method described in embodiment 1-3:Phoenix-tail fern is crushed, crosses 40-50 mesh
Sieve, weighs powder 40-50g, adds refluxing extraction 3-5 times at 60-70 DEG C of the 70-90% ethanol of 2-4 times of volume, 1.5-2h/ times,
Merging filtrate, centrifugation, takes supernatant, and it is alcohol extract to volatilize after solvent, alcohol extract is dissolved with absolute ethyl alcohol, after distilled water diluting
PH is adjusted to neutrality, 4 DEG C save backup after sterilizing.
Using conventional salt tolerant, high temperature resistant method detection embodiment 1-3 and the cepe of comparative example 1 expand breeding 50,
100th, 150, the vigor under 200mmol/LNaCl;And the vigor at a temperature of 30,35,40 DEG C.It the results are shown in Table 1, table 2.
The embodiment 1-3 of table 1 and the cepe of comparative example 1 expand breeding salt tolerant test result
As known from Table 1 compared with expanding in breeding culture medium and not adding the comparative example 1 of phoenix-tail fern extract solution, embodiment 1-3 is delicious
The vigor that bolete expands breeding significantly improves, it is seen that phoenix-tail fern extract solution can improve the salt tolerant energy that cepe expands breeding
Power.
The embodiment 1-3 of table 2 and the cepe of comparative example 1 expand breeding high temperature resistant test result
As known from Table 2 compared with expanding in breeding culture medium and not adding the comparative example 1 of phoenix-tail fern extract solution, embodiment 1-3 is delicious
The vigor that bolete expands breeding significantly improves, it is seen that phoenix-tail fern extract solution can improve the high temperature resistant energy that cepe expands breeding
Power.
The nutritional ingredient of cepe obtained by embodiment 1-3 fruitings is detected, and is sought with the cepe of field acquisition
Form and be divided into reference, the results are shown in Table 3.
The embodiment 1-3 fruitings of table 3 gained and the cepe nutrient component meter of field acquisition
As known from Table 3, the nutritional ingredient protein of cepe obtained by 1-3 of embodiment of the present invention fruitings, carbon hydrate
Thing, heat, ash content, Ca, P, Fe, riboflavin are above the content of the corresponding nutritional ingredient of cepe of field acquisition.
Obviously, above-described embodiment is only the section Example of the present invention, rather than whole embodiments.Based on opening for the present invention
Show, those of ordinary skill in the art's all other embodiment resulting on the premise of creative work is not made, all
Belong to the scope that present aspect is protected.
Claims (6)
1. a kind of artificial cultivation method of cepe, it is characterised in that comprise the following steps:
A. parent species are separately cultured:Fresh, sturdy, medium ripe wild fructification is gathered, is cleaned, surface sterilization, in sterile bar
Tissue block is cut under part to cultivate in mother culture media;
B. strain expands numerous:Step a is cultivated into the access of gained parent species is containing shaken cultivation in the container for expanding breeding culture medium, frequency of oscillation
10-30rpm/min;
C. lophatherum gracile cultivating aseptic seedling:Ripe lophatherum gracile bamboo rice is selected, sterilization of shelling is placed on inducing culture after rinsing and cultivated,
After callus is induced lophatherum gracile regeneration plant i.e. lophatherum gracile aseptic seedling is obtained through subculture and transdifferentiation culture;
D. mycorrhiza fungi seeding cultivating and cultivation fruiting:The bacterium ball for expanding numerous acquisition is broken into bacterium slurry, 10-50 times is diluted, is injected into henon bamboo
Va Mycorrhiza Seedling up to being trained Va Mycorrhiza Seedling, is then implanted into earthing mushroom bed by cultivation again after leaf aseptic seedling, cultivates fruiting.
2. artificial cultivation method according to claim 1, it is characterised in that methods described step a tissue block is stem, bacterium
Lid or cap and stem intersection.
3. artificial cultivation method according to claim 1, it is characterised in that methods described step a mother culture media is horse
Bell potato 160-180g, glucose 15-17g, Yeast diffusion juice 1.6-1.8g, vitamin B126-32ug, water 900-1000ml.
4. artificial cultivation method according to claim 1, it is characterised in that methods described step b expansion breeding culture medium is dark
Brown earth 4-4.5kg, wheat 4-4.5kg, birch wood chip 1.5-2.5kg, turf 0.6-1kg, potassium sulfate 0.1-0.2kg, peroxophosphoric acid
Calcium 0.1-0.2kg, phoenix-tail fern extract solution 3-7ml.
5. artificial cultivation method according to claim 4, it is characterised in that the extracting method of the phoenix-tail fern extract solution is will
Phoenix-tail fern crushes.40-50 mesh sieves are crossed, weigh powder 40-50g, adds at 60-70 DEG C of the 70-90% ethanol of 2-4 times of volume and flows back
Extraction 3-5 time, 1.5-2h/ times, merging filtrate, centrifuge, take supernatant, it is alcohol extract to volatilize after solvent, by alcohol extract with anhydrous
Ethanol dissolves, and pH is adjusted after distilled water diluting to neutrality, 4 DEG C save backup after sterilizing.
6. according to any one of the claim 1-5 artificial cultivation methods, it is characterised in that methods described step b earthing mushroom
The soil used in bed is scpiolitc powder, grey sandy soil, dark brown earth are with 1:2-3:2.5-4 ratio mix.
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Citations (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN101204131A (en) * | 2007-12-10 | 2008-06-25 | 丽水市林业科学研究所 | Artificial cultivation method for Suillus Luteus |
CN101669430A (en) * | 2009-09-24 | 2010-03-17 | 云南省热带作物科学研究所 | Semi-artificial culture method of phlebopus portentosus |
CN104892095A (en) * | 2015-05-08 | 2015-09-09 | 云南省热带作物科学研究所 | Mould-inhibition and yield-increasing panacea dedicated to soil covering process for Phlebopus portentosus |
CN105418265A (en) * | 2015-12-24 | 2016-03-23 | 桂林大野领御生物科技有限公司 | Culture medium for substitute cultivation of ganoderma applanatum |
-
2017
- 2017-08-23 CN CN201710738731.XA patent/CN107360859A/en active Pending
Patent Citations (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN101204131A (en) * | 2007-12-10 | 2008-06-25 | 丽水市林业科学研究所 | Artificial cultivation method for Suillus Luteus |
CN101669430A (en) * | 2009-09-24 | 2010-03-17 | 云南省热带作物科学研究所 | Semi-artificial culture method of phlebopus portentosus |
CN104892095A (en) * | 2015-05-08 | 2015-09-09 | 云南省热带作物科学研究所 | Mould-inhibition and yield-increasing panacea dedicated to soil covering process for Phlebopus portentosus |
CN105418265A (en) * | 2015-12-24 | 2016-03-23 | 桂林大野领御生物科技有限公司 | Culture medium for substitute cultivation of ganoderma applanatum |
Non-Patent Citations (1)
Title |
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潘崇环等: "《珍稀食用菌栽培与名贵野生菌的开发利用》", 31 January 2006, 中国农业出版社 * |
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