CN105349436A - Preparation method of wild tricholoma matsutake domesticated mother strains - Google Patents
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Abstract
The invention discloses a preparation method of wild tricholoma matsutake domesticated mother strains. The method comprises the following steps that tricholoma matsutake which grows in the wild naturally is collected to prepare a spore solution; the spore solution is added to a liquid culture medium to conduct bacterium shaking, after liquid multi-spore selfing is conducted, the obtained tricholoma matsutake mixed hypha liquid is inoculated on a culture material of a cultivation bottle in a clean room, sealing is conducted through a vent cap, the cultivation bottle is put in a culture room, dark culture is conducted for 25-30 d, and ultrasonic water bath culture is conducted on 2-6 d; when hyphae grow all over the cultivation bottle, infection treatment is conducted on dominant associated tree species of the tricholoma matsutake, 900 ml-1000 ml of water is used for watering, mulch is used for covering, artificial domestication is conducted, and the tricholoma matsutake mother strains which are high in stability and capable of being used for breeding and production can be obtained. According to the preparation method of the wild tricholoma matsutake domesticated mother strains, by means of reasonable improvement of the culture medium and the culture method, artificial domestication of the tricholoma matsutake mother strains is achieved, the current situation that tricholoma matsutake production completely relies on nature is changed, and stable yield, high yield and high quality of tricholoma matsutake products are further achieved.
Description
Technical field
The present invention relates to matsutake and cultivate field, be specifically related to a kind of wild matsutake domestication female kind preparation method.
Background technology
Matsutake TricholomamatsutakeSing. is one of world-renowned wild edible fungus.Principal market, the matsutake Yi Zhan world---more than 60% of the Japanese market sales volume of Yunnan outlet in recent years, year outlet matsutake is earned foreign exchange and is reached more than 4,000 ten thousand dollars, and within 2004, reach 5,361 ten thousand dollars, export volume is 1280.85 tons, wherein fresh matsutake 973.5 tons, 307.35 tons, matsutake goods.According to investigations, Deqin County and Xianggelila country peasant economy income more than 60% derive from matsutake.The continuing of industry chain of Tricholoma spp, develop in a healthy way shaking off poverty and setting out on the road to prosperity for hill farmer, the development of the rural economy of border ethnic minorities area, the stable of society provide Important Economic guarantee.
The matsutake domestication research in century more than one shows, directly adopting Pure cultured spawn to realize matsutake artificial culture is a kind of target being difficult to realize, and for this reason, is necessary very much to carry out artificial domesticating cultivation technological development.
Summary of the invention
For solving the problem, the invention provides a kind of wild matsutake domestication female kind preparation method.
For achieving the above object, the technical scheme that the present invention takes is:
Wild matsutake domestication is female plants preparation method, comprises the steps:
The matsutake of S1, collection nature field grown, get full grown sporophore cap part, cut away stem, cotton ball soaked in alcohol with 75% cleans cap surface, lamella catches on sterilized cleek down and hangs in the triangular flask of sterilizing, spend the night with aseptic tampon sealing, collect spore print, make spore liquid with sterilized water;
S2, join in liquid nutrient medium and carry out shaking bacterium, carry out the selfing of liquid many spores, shaking bacterium condition is: rotating speed 140r/min, and culture temperature is 25 DEG C, obtains matsutake mixing mycelia liquid;
S3, the matsutake of gained mixing mycelia liquid is inoculated on the culture material of culture bottle in clean room, seal with ventilating cover, and light culture 25 ~ 30d in the culturing room being placed in 14 ~ 15 DEG C, when 2 ~ 6d, carry out ultrasound bath cultivation, hyperacoustic frequency and power are set as 68kHz and 600W respectively, and ultrasonic treatment time is 10min ~ 90min, and ultrasonic interval is than being 20s/5s ~ 30s/5s;
S4, cover with after culture bottle until mycelia, infect the advantage associated species of process matsutake, water 900ml ~ 1000ml, builds coverture, domestication, can obtain stability strong, and the matsutake mother that can be used for propagation production plants.
Wherein, the liquid nutrient medium in described step S2 often rises and contains:
Glucose 20 ~ 24g, yeast extract paste 0.3 ~ 0.5g, peptone 0.5 ~ 0.7g, Nostoc lyophilized powder 3 ~ 5g, amino acid chelated magnesium 0.05 ~ 0.07g, potassium primary phosphate 0.1 ~ 0.2g, water soluble starch 40 ~ 60g, all the other are sterilized water.
Wherein, the culture material in described step S3 is prepared by following steps:
(1) take pine tree wood chip 36 ~ 48 parts, Rhizoma amorphophalli powder 8 ~ 10g, garlicin 0.5 ~ 0.6 part, weathered coal 8 ~ 10 parts be placed in steam-explosion jar, first passing into nitrogen to steam-explosion jar internal pressure is 0.7 ~ 1.3MPa, explosion treatment 8 ~ 25min; Then passing into rapidly steam to steam-explosion jar internal pressure is 1.3 ~ 1.7MPa, and Steam explosion treatment 0.5 ~ 2.5min, obtains mixture;
(2) by the mixture Temperature fall of gained to normal temperature, add silicon nitride super-fine powder 3 ~ 7 parts, chitin fiber 4 ~ 6 parts, alginate fiber 3 ~ 5 parts of mixing and stirring, and be 64 ~ 68% of gross weight by adding water adjustment moisture content, obtain culture base-material for subsequent use.
Wherein, described Nostoc lyophilized powder is prepared by following steps:
Get 50 ~ 55 portions of Nostoc, cleaning, wears into meal with shredder, then uses 90 ~ 95 DEG C of hot-water soak 50 ~ 85min, pull out after draining, add 10 ~ 12 parts of normal temperature pure water, and even, then according to the ratio microbe inoculation starter of inoculum size 1 ~ 1.5%, keep temperature 38 DEG C, ferment 52 ~ 58 hours, oven drying at low temperature after fermentation ends, grinds to form fine powder.
Wherein, the composition of described microbial starter culture is: according to bacterial strain number ratio lactobacillus: Bacillus licheniformis: Corynebacterium glutamicum is: the proportions of 1: 1.2: 2 becomes 1.5 × 10
7the starter of bacterial strain/ml.
The present invention has following beneficial effect:
By the rational modification of substratum and cultural method, achieve the domestication that matsutake mother plants, change matsutake and produce and rely on natural present situation completely, but also achieve stable and high yields and the high-quality of matsutake product.Meanwhile, the ecotope that the present invention also effectively protects matsutake germ plasm resource and relies on, the Sustainable development of producing for achieving matsutake provides an effective approach.
Embodiment
In order to make objects and advantages of the present invention clearly understand, below in conjunction with embodiment, the present invention is further elaborated.Should be appreciated that specific embodiment described herein only in order to explain the present invention, be not intended to limit the present invention.
In below implementing, the liquid nutrient medium in described step S2 often rises and contains:
Glucose 20 ~ 24g, yeast extract paste 0.3 ~ 0.5g, peptone 0.5 ~ 0.7g, Nostoc lyophilized powder 3 ~ 5g, amino acid chelated magnesium 0.05 ~ 0.07g, potassium primary phosphate 0.1 ~ 0.2g, water soluble starch 40 ~ 60g, all the other are sterilized water.
Culture material in described step S3 is prepared by following steps:
(1) take pine tree wood chip 36 ~ 48 parts, Rhizoma amorphophalli powder 8 ~ 10g, garlicin 0.5 ~ 0.6 part, weathered coal 8 ~ 10 parts be placed in steam-explosion jar, first passing into nitrogen to steam-explosion jar internal pressure is 0.7 ~ 1.3MPa, explosion treatment 8 ~ 25min; Then passing into rapidly steam to steam-explosion jar internal pressure is 1.3 ~ 1.7MPa, and Steam explosion treatment 0.5 ~ 2.5min, obtains mixture;
(2) by the mixture Temperature fall of gained to normal temperature, add silicon nitride super-fine powder 3 ~ 7 parts, chitin fiber 4 ~ 6 parts, alginate fiber 3 ~ 5 parts of mixing and stirring, and be 64 ~ 68% of gross weight by adding water adjustment moisture content, obtain culture base-material for subsequent use.
Described Nostoc lyophilized powder is prepared by following steps:
Get 50 ~ 55 portions of Nostoc, cleaning, wears into meal with shredder, then uses 90 ~ 95 DEG C of hot-water soak 50 ~ 85min, pull out after draining, add 10 ~ 12 parts of normal temperature pure water, and even, then according to the ratio microbe inoculation starter of inoculum size 1 ~ 1.5%, keep temperature 38 DEG C, ferment 52 ~ 58 hours, oven drying at low temperature after fermentation ends, grinds to form fine powder.
Wherein, the composition of described microbial starter culture is: according to bacterial strain number ratio lactobacillus: Bacillus licheniformis: Corynebacterium glutamicum is: the proportions of 1: 1.2: 2 becomes 1.5 × 10
7the starter of bacterial strain/ml.
Embodiment 1
The matsutake of S1, collection nature field grown, get full grown sporophore cap part, cut away stem, cotton ball soaked in alcohol with 75% cleans cap surface, lamella catches on sterilized cleek down and hangs in the triangular flask of sterilizing, spend the night with aseptic tampon sealing, collect spore print, make spore liquid with sterilized water;
S2, join in liquid nutrient medium and carry out shaking bacterium, carry out the selfing of liquid many spores, shaking bacterium condition is: rotating speed 140r/min, and culture temperature is 25 DEG C, obtains matsutake mixing mycelia liquid;
S3, the matsutake of gained mixing mycelia liquid is inoculated on the culture material of culture bottle in clean room, seal with ventilating cover, and light culture 25d in the culturing room being placed in 14 ~ 15 DEG C, when 2d, carry out ultrasound bath cultivation, hyperacoustic frequency and power are set as 68kHz and 600W respectively, and ultrasonic treatment time is that the ultrasonic interval of 10min is than being 20s/5s;
S4, cover with after culture bottle until mycelia, infect the advantage associated species of process matsutake, water 900ml, builds coverture, domestication, can obtain stability strong, and the matsutake mother that can be used for propagation production plants.
Embodiment 2
The matsutake of S1, collection nature field grown, get full grown sporophore cap part, cut away stem, cotton ball soaked in alcohol with 75% cleans cap surface, lamella catches on sterilized cleek down and hangs in the triangular flask of sterilizing, spend the night with aseptic tampon sealing, collect spore print, make spore liquid with sterilized water;
S2, join in liquid nutrient medium and carry out shaking bacterium, carry out the selfing of liquid many spores, shaking bacterium condition is: rotating speed 140r/min, and culture temperature is 25 DEG C, obtains matsutake mixing mycelia liquid;
S3, the matsutake of gained mixing mycelia liquid is inoculated on the culture material of culture bottle in clean room, seal with ventilating cover, and light culture 30d in the culturing room being placed in 15 DEG C, when 6d, carry out ultrasound bath cultivation, hyperacoustic frequency and power are set as 68kHz and 600W respectively, and ultrasonic treatment time is 90min, and ultrasonic interval is than being 30s/5s;
S4, cover with after culture bottle until mycelia, infect the advantage associated species of process matsutake, water 1000ml, builds coverture, domestication, can obtain stability strong, and the matsutake mother that can be used for propagation production plants.
Embodiment 3
The matsutake of S1, collection nature field grown, get full grown sporophore cap part, cut away stem, cotton ball soaked in alcohol with 75% cleans cap surface, lamella catches on sterilized cleek down and hangs in the triangular flask of sterilizing, spend the night with aseptic tampon sealing, collect spore print, make spore liquid with sterilized water;
S2, join in liquid nutrient medium and carry out shaking bacterium, carry out the selfing of liquid many spores, shaking bacterium condition is: rotating speed 140r/min, and culture temperature is 25 DEG C, obtains matsutake mixing mycelia liquid;
S3, the matsutake of gained mixing mycelia liquid is inoculated on the culture material of culture bottle in clean room, seal with ventilating cover, and light culture 27.5d in the culturing room being placed in 14.5 DEG C, when 4d, carry out ultrasound bath cultivation, hyperacoustic frequency and power are set as 68kHz and 600W respectively, and ultrasonic treatment time is 50min, and ultrasonic interval is than being 25s/5s;
S4, cover with after culture bottle until mycelia, infect the advantage associated species of process matsutake, water 950ml, builds coverture, domestication, can obtain stability strong, and the matsutake mother that can be used for propagation production plants.
The above is only the preferred embodiment of the present invention; it should be pointed out that for those skilled in the art, under the premise without departing from the principles of the invention; can also make some improvements and modifications, these improvements and modifications also should be considered as protection scope of the present invention.
Claims (5)
1. wild matsutake domestication is female plants preparation method, it is characterized in that, comprises the steps:
The matsutake of S1, collection nature field grown, get full grown sporophore cap part, cut away stem, cotton ball soaked in alcohol with 75% cleans cap surface, lamella catches on sterilized cleek down and hangs in the triangular flask of sterilizing, spend the night with aseptic tampon sealing, collect spore print, make spore liquid with sterilized water;
S2, join in liquid nutrient medium and carry out shaking bacterium, carry out the selfing of liquid many spores, shaking bacterium condition is: rotating speed 140r/min, and culture temperature is 25 DEG C, obtains matsutake mixing mycelia liquid;
S3, the matsutake of gained mixing mycelia liquid is inoculated on the culture material of culture bottle in clean room, seal with ventilating cover, and light culture 25 ~ 30d in the culturing room being placed in 14 ~ 15 DEG C, when 2 ~ 6d, carry out ultrasound bath cultivation, hyperacoustic frequency and power are set as 68kHz and 600W respectively, and ultrasonic treatment time is 10min ~ 90min, and ultrasonic interval is than being 20s/5s ~ 30s/5s;
S4, cover with after culture bottle until mycelia, infect the advantage associated species of process matsutake, water 900ml ~ 1000ml, builds coverture, domestication, can obtain stability strong, and the matsutake mother that can be used for propagation production plants.
2. wild matsutake domestication according to claim 1 is female plants preparation method, and it is characterized in that, the liquid nutrient medium in described step S2 often rises and contains:
Glucose 20 ~ 24g, yeast extract paste 0.3 ~ 0.5g, peptone 0.5 ~ 0.7g, Nostoc lyophilized powder 3 ~ 5g, amino acid chelated magnesium 0.05 ~ 0.07g, potassium primary phosphate 0.1 ~ 0.2g, water soluble starch 40 ~ 60g, all the other are sterilized water.
3. wild matsutake domestication according to claim 1 is female plants preparation method, and it is characterized in that, the culture material in described step S3 is prepared by following steps:
(1) take pine tree wood chip 36 ~ 48 parts, Rhizoma amorphophalli powder 8 ~ 10g, garlicin 0.5 ~ 0.6 part, weathered coal 8 ~ 10 parts be placed in steam-explosion jar, first passing into nitrogen to steam-explosion jar internal pressure is 0.7 ~ 1.3MPa, explosion treatment 8 ~ 25min; Then passing into rapidly steam to steam-explosion jar internal pressure is 1.3 ~ 1.7MPa, and Steam explosion treatment 0.5 ~ 2.5min, obtains mixture;
(2) by the mixture Temperature fall of gained to normal temperature, add silicon nitride super-fine powder 3 ~ 7 parts, chitin fiber 4 ~ 6 parts, alginate fiber 3 ~ 5 parts of mixing and stirring, and be 64 ~ 68% of gross weight by adding water adjustment moisture content, obtain culture base-material for subsequent use.
4. wild matsutake domestication according to claim 1 is female plants preparation method, and it is characterized in that, described Nostoc lyophilized powder is prepared by following steps:
Get 50 ~ 55 portions of Nostoc, cleaning, wears into meal with shredder, then uses 90 ~ 95 DEG C of hot-water soak 50 ~ 85min, pull out after draining, add 10 ~ 12 parts of normal temperature pure water, and even, then according to the ratio microbe inoculation starter of inoculum size 1 ~ 1.5%, keep temperature 38 DEG C, ferment 52 ~ 58 hours, oven drying at low temperature after fermentation ends, grinds to form fine powder.
5. wild matsutake domestication according to claim 4 is female plants preparation method, it is characterized in that, the composition of described microbial starter culture is: according to bacterial strain number ratio lactobacillus: Bacillus licheniformis: Corynebacterium glutamicum is: the proportions of 1: 1.2: 2 becomes 1.5 × 10
7the starter of bacterial strain/ml.
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105900620A (en) * | 2016-03-28 | 2016-08-31 | 河南科技学院 | Cucumber planting method |
| CN106212042A (en) * | 2016-07-04 | 2016-12-14 | 李业武 | A kind of pseudo-wild cultivating method of Tricholoma matsutake (lto et lmai) Singer |
| CN108293614A (en) * | 2016-09-27 | 2018-07-20 | 陈晓红 | The method of matsutake edible fungus industrial production |
| CN109735465A (en) * | 2019-01-25 | 2019-05-10 | 江苏大学 | A kind of microbial preparation for improving plant growth on tidal flat and preparation method thereof |
| CN112831456A (en) * | 2019-11-25 | 2021-05-25 | 湖南金芙农业科技有限公司 | Tricholoma matsutake sexual spore and separation method thereof |
Citations (1)
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| CN103650912A (en) * | 2013-11-26 | 2014-03-26 | 华中农业大学 | Multi-spore selfing breeding method of golden needle mushroom |
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| CN103650912A (en) * | 2013-11-26 | 2014-03-26 | 华中农业大学 | Multi-spore selfing breeding method of golden needle mushroom |
Non-Patent Citations (4)
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| SUNG SU KIM ET AL.: "Process development for mycelial growth and polysaccharide production in Tricholoma matsutake liquid culture", 《JOURNAL OF BIOSCIENCE AND BIOENGINEERING》 * |
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105900620A (en) * | 2016-03-28 | 2016-08-31 | 河南科技学院 | Cucumber planting method |
| CN106212042A (en) * | 2016-07-04 | 2016-12-14 | 李业武 | A kind of pseudo-wild cultivating method of Tricholoma matsutake (lto et lmai) Singer |
| CN108293614A (en) * | 2016-09-27 | 2018-07-20 | 陈晓红 | The method of matsutake edible fungus industrial production |
| CN109735465A (en) * | 2019-01-25 | 2019-05-10 | 江苏大学 | A kind of microbial preparation for improving plant growth on tidal flat and preparation method thereof |
| CN112831456A (en) * | 2019-11-25 | 2021-05-25 | 湖南金芙农业科技有限公司 | Tricholoma matsutake sexual spore and separation method thereof |
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