CN108901589A - A kind of separation method of wild delicious lactarius strain - Google Patents
A kind of separation method of wild delicious lactarius strain Download PDFInfo
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- CN108901589A CN108901589A CN201810351695.6A CN201810351695A CN108901589A CN 108901589 A CN108901589 A CN 108901589A CN 201810351695 A CN201810351695 A CN 201810351695A CN 108901589 A CN108901589 A CN 108901589A
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- strain
- delicious lactarius
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- separation method
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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
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G18/00—Cultivation of mushrooms
- A01G18/20—Culture media, e.g. compost
Abstract
The present invention relates to a kind of separation methods of wild delicious lactarius strain.The separation method of the wild delicious lactarius strain, which is characterized in that including：It acquires wild delicious lactarius fructification and separates lamella, cultivated in the first culture medium, the strain of acquisition is purified, then go to ordinary culture medium and carry out adaptability culture, obtain wild delicious lactarius strain.The advantages that wild delicious lactarius strain that the present invention obtains is active strong, and the speed of growth is fast, it is unique in the wild delicious lactarius strain method in China for separating.
Description
Technical field
The invention belongs to the acquisition of delicious lactarius strain and application fields, more particularly to the wild delicious lactarius strain using Zhejiang
It obtains and applies.
Background technique
Delicious lactarius (Lactarius deliciosus) belongs to Russulaceae, Lactarius, also known as delicious delicious lactarius, pine tree mushroom, pine
Bacterium, fir tree mushroom, cogongrass mushroom are a kind of well received delicious edible mushrooms.China is that wild mushroom bio-diversity is very rich
One of country, China are estimated to be 100,000 kinds of edible and medical fungis, only 1000 kinds or so be realized, and only 100 be utilized
Kind or so, it can be seen that most of wild edible and medical fungis are undiscovered, recognize and utilize.The better edible and medical fungi of domesticating and cultivating is
One of the important process of edible and medical fungi worker.
The whole nation such as China Zhejiang, Hunan, Hubei, Anhui, Guangxi, Guizhou, Yunnan, Guangdong most area has delicious lactarius
Distribution, and welcome in Central China, south China and East China some areas by common people.But delicious lactarius strain is difficult to be separated at present
Function, it is not strong separating successfully its speed of growth or activity, therefore much tamed by the edible mushroom of research object of delicious lactarius
It has no idea to carry out with cultivation activity.
In face of this situation, finding one kind can be with quick separating delicious lactarius strain, and delicious lactarius mycelia can be made commonly to train
Fast-growth on base is supported, is the prerequisite for carrying out delicious lactarius research.
Summary of the invention
The object of the present invention is to provide a kind of separation methods of wild delicious lactarius strain for originating from Zhejiang.
In order to achieve the above object, the present invention provides a kind of separation methods of wild delicious lactarius strain, which is characterized in that
Including：It acquires wild delicious lactarius (Lactarius deliciosus) fructification and separates lamella, cultivated in the first culture medium,
The strain of acquisition is purified, then goes to ordinary culture medium and carries out adaptability culture, obtains wild delicious lactarius strain；Wherein,
First culture medium contain 18-22g/L Fagaceae water extract, 18-22g/L agar, 18-22g/L glucose,
0.05-0.15g/L calcium chloride, 0.05-0.15g/L magnesium sulfate, 0.5-1.0g/L potassium dihydrogen phosphate, 0.005-0.015g/L chlorination
Sodium, 0.1-0.3g/L dihydrogen phosphate, 0.05-0.15g/L iron chloride, 0.05-0.15g/L zinc sulfate, 0.03-0.08g/L sulphur
Sour copper, 0.005-0.015g/L niacin, 0.05-0.15mg/L vitamin B1And 0.5-1.5g/L brewer's wort, the shell
The preparation method of bucket section low cost vegetable plant water extract includes：The withered blade of Fagaceae is boiled with water, Fagaceae dries up
The weight ratio of blade and water is 1: 80-120, filtering, and gained filtrate is Fagaceae water extract.
Preferably, the Fagaceae is Chinese chestnut (Castanea mollissima).
Preferably, the tanning time is 5-15min.
Preferably, the preparation method of first culture medium includes：Agar is added simultaneously in Fagaceae water extract
Heating and melting, is added drug after agar melts completely, and the drug includes glucose, calcium chloride, magnesium sulfate, biphosphate
Potassium, sodium chloride, dihydrogen phosphate, iron chloride, zinc sulfate, copper sulphate, niacin, vitamin B1And brewer's wort, it is complete to drug
After full thawing, constant volume sterilizes, packing, condensation to room temperature.
Wild delicious lactarius of the invention is grown on Zhejiang.
Preferably, the wild delicious lactarius is acquired from Fagaceae tree is lower.
Preferably, the method for acquiring wild delicious lactarius fructification and separating lamella includes：It acquires wild fresh
Delicious lactarius fructification removes surface irregularities and dust, chooses the lamella being not exposed in air and is separated.
Preferably, the condition cultivated in the first culture medium is to cultivate 10-15d under 20-30 DEG C of half-light.
Preferably, the step of purifying agaric includes：The strain of acquisition is inoculated in the second culture medium and is trained
Support the growth to inhibit bacterium, the second culture medium 15-25g/L containing glucose, yeast extract 1-3g/L, citric acid 0.3-
0.8g/L, agar 15-25g/L, condition of culture are that room temperature is protected from light culture.
Preferably, the ordinary culture medium is PDA culture medium, and condition of culture is half-light culture 5-10d at 20-30 DEG C.
Preferably, the separation method of the wild delicious lactarius strain further includes：To the delicious lactarius pure culture of acquisition and point
From delicious lactarius fructification lamella carry out DNA sequence dna sequencing, the sequence of acquisition is compared, sequence 100% unanimously then obtains
Delicious lactarius pure culture be pure culture.
The present invention efficiently separates a kind of wild delicious lactarius strain for being grown on Zhejiang using the culture medium of formulated, warp
This extraordinary culture medium and this method carry out lamella separation, are transferred to ordinary culture medium progress adaptability training again after growing mycelia
It supports, the strain after adaptation carries out microstructure and molecular sequence identification to be confirmed whether the strain of the delicious lactarius, isolated strain
It can be used for the subsequent domesticating and cultivating etc. of delicious lactarius.
It joined the withered blade of Fagaceae of tanning in special culture medium in the present invention, while joined in drug
Zinc sulfate, copper sulphate, niacin ingredient, these ingredients are verified through test of many times, can make the wild delicious lactarius bacterium for originating from Zhejiang
Silk fast-germination after inoculation.
Lamella (the size about 2 × 2mm of the present invention being not exposed in air2), rather than meat bacteria organization or ejection
Spore, avoid pollution, spore content is high, is easy to sprout.
Compared with prior art, the beneficial effects of the invention are as follows：
The present invention can quick separating delicious lactarius pure culture, separated delicious lactarius pure culture can fast-growth, can be used for
Research and domesticating and cultivating.
Detailed description of the invention
Fig. 1 separates successfully wild delicious lactarius strain.
Specific embodiment
Present invention will be further explained below with reference to specific examples.It should be understood that these embodiments are merely to illustrate the present invention
Rather than it limits the scope of the invention.In addition, it should also be understood that, after reading the content taught by the present invention, those skilled in the art
Member can make various changes or modifications the present invention, and such equivalent forms equally fall within the application the appended claims and limited
Range.
Embodiment 1
A kind of separation method of wild delicious lactarius strain, including：
(1) first culture medium is prepared：First culture medium contains 20g/L Fagaceae Chinese chestnut water extract, 20g/L
Agar, 20g/L glucose, 0.1g/L calcium chloride, 0.1g/L magnesium sulfate, 0.8g/L potassium dihydrogen phosphate, 0.01g/L sodium chloride,
0.2g/L dihydrogen phosphate, 0.1g/L iron chloride, 0.1g/L zinc sulfate, 0.05g/L copper sulphate, 0.01g/L niacin, 0.1mg/
L vitamin B1And 1g/L brewer's wort, preparation method are：The withered blade of Fagaceae Chinese chestnut is added into water, acorn-cup
The withered blade of section plant Chinese chestnut and the weight ratio of water are 1: 100, are filtered after boiling 10min, and gained filtrate is Fagaceae
Water extract, agar is added in filtrate in proportion and heats makes its thawing, sequentially adds drug, drug after agar melts completely
Including glucose, calcium chloride, magnesium sulfate, potassium dihydrogen phosphate, sodium chloride, dihydrogen phosphate, iron chloride, zinc sulfate, copper sulphate, Buddhist nun
Gram acid, vitamin B1, brewer's wort (Qingdao HB4176-3 Hai Bosheng Technology Co., Ltd.), after drug melts completely, be settled to
1000ml；121 DEG C of sterilizing 15min, it is rear to dispense test tube or plate, it condenses spare to room temperature.
(2) strain separating and culture：Wild fresh delicious lactarius fructification is acquired under the Fagaceae tree Chinese chestnut of Zhejiang,
Surface irregularities and dust are removed, lamella (the size about 2 × 2mm being not exposed in air is chosen2) separated, it is placed in and has gone out
In the first culture medium test tube or plate of bacterium, 10-15d is cultivated under 25 DEG C of half-lights, obtains delicious lactarius strain.
(3) strain of acquisition is purified：The strain of acquisition is inoculated in the second culture medium and is cultivated to inhibit
The growth of bacterium, the second culture medium 20g/L containing glucose, yeast extract 2g/L, citric acid 0.5g/L, agar 20g/L, training
The condition of supporting is that room temperature is protected from light culture, obtains the pure culture of delicious lactarius.
(4) ordinary culture medium tube adapts to：Picking purifying delicious lactarius mycelia, tube to common PDA culture medium test tube or
Adaptability culture is carried out on plate, half-light culture 5-10d at 25 DEG C obtains delicious lactarius pure culture.
(5) strain idenfication：Delicious lactarius pure culture and isolated delicious lactarius fructification lamella to acquisition carry out DNA sequence dna survey
The sequence of acquisition is compared sequence, and the strain that sequence 100% unanimously then obtains is pure culture.The present invention can be isolated and be adopted
Collect the delicious lactarius strain from Zhejiang, other then cannot.
Claims (10)
1. a kind of separation method of wild delicious lactarius strain, which is characterized in that including：It acquires wild delicious lactarius fructification and separates
Lamella is cultivated in the first culture medium, and the strain of acquisition is purified, then is gone to ordinary culture medium and carried out adaptability culture,
Obtain wild delicious lactarius strain；Wherein, first culture medium contains 18-22g/L Fagaceae water extract, 18-22g/L
Agar, 18-22g/L glucose, 0.05-0.15g/L calcium chloride, 0.05-0.15g/L magnesium sulfate, 0.5-1.0g/L biphosphate
Potassium, 0.005-0.015g/L sodium chloride, 0.1-0.3g/L dihydrogen phosphate, 0.05-0.15g/L iron chloride, 0.05-0.15g/L
Zinc sulfate, 0.03-0.08g/L copper sulphate, 0.005-0.015g/L niacin, 0.05-0.15mg/L vitamin B1And 0.5-
The preparation method of 1.5g/L brewer's wort, the Fagaceae water extract includes：By the withered blade water of Fagaceae
Tanning, the withered blade of Fagaceae and the weight ratio of water are 1: 80-120, filtering, and gained filtrate is that Fagaceae water mentions
Object.
2. the separation method of wild delicious lactarius strain as described in claim 1, which is characterized in that the Fagaceae is
Chinese chestnut.
3. the separation method of wild delicious lactarius strain as described in claim 1, which is characterized in that the tanning time is 5-
15min。
4. the separation method of wild delicious lactarius strain as described in claim 1, which is characterized in that first culture medium
Preparation method includes：Agar and heating and melting are added in Fagaceae water extract, drug is added after agar melts completely,
The drug include glucose, calcium chloride, magnesium sulfate, potassium dihydrogen phosphate, sodium chloride, dihydrogen phosphate, iron chloride, zinc sulfate,
Copper sulphate, niacin, vitamin B1And brewer's wort, after drug melts completely, constant volume sterilizes, packing, condensation to room temperature.
5. the separation method of wild delicious lactarius strain as described in claim 1, which is characterized in that the wild delicious lactarius from
Fagaceae tree is lower to be acquired.
6. the separation method of wild delicious lactarius strain as described in claim 1, which is characterized in that the wild pine cream of the acquisition
The massee fruiting bodies and method for separating lamella includes：Wild fresh delicious lactarius fructification is acquired, surface irregularities and dust, choosing are removed
The lamella being not exposed in air is taken to be separated.
7. the separation method of wild delicious lactarius strain as described in claim 1, which is characterized in that described in the first culture medium
The condition of middle culture is to cultivate 10-15d under 20-30 DEG C of half-light.
8. the separation method of wild delicious lactarius strain as described in claim 1, which is characterized in that the step of the purifying agaric
Suddenly include：The strain of acquisition is inoculated in the growth cultivated in the second culture medium to inhibit bacterium, second culture
Base 15-25g/L containing glucose, yeast extract 1-3g/L, citric acid 0.3-0.8g/L, agar 15-25g/L, condition of culture are room temperature
It is protected from light culture.
9. the separation method of wild delicious lactarius strain as described in claim 1, which is characterized in that the ordinary culture medium is
PDA culture medium, condition of culture are half-light culture 5-10d at 20-30 DEG C.
10. the separation method of wild delicious lactarius strain as described in claim 1, which is characterized in that the wild delicious lactarius
The separation method of strain further includes：Delicious lactarius pure culture and isolated delicious lactarius fructification lamella to acquisition carry out DNA sequence dna
Sequencing, the sequence of acquisition is compared, and the delicious lactarius pure culture that sequence 100% unanimously then obtains is pure culture.
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Cited By (1)
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| CN114317279A (en) * | 2021-12-20 | 2022-04-12 | 贵州省生物研究所 | High-quality culture medium formula for separating pleurotus nebrodensis associated with fungi |
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| WO2023116262A1 (en) * | 2021-12-20 | 2023-06-29 | 贵州省生物研究所 | Formula of high-quality culture medium for isolating accompanying calcarisporium arbuscula fungus of lactarius lignyotus |
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