CN106613350A - Schizophyllum commune and domestication and cultivation method and application thereof - Google Patents
Schizophyllum commune and domestication and cultivation method and application thereof Download PDFInfo
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- CN106613350A CN106613350A CN201611193054.XA CN201611193054A CN106613350A CN 106613350 A CN106613350 A CN 106613350A CN 201611193054 A CN201611193054 A CN 201611193054A CN 106613350 A CN106613350 A CN 106613350A
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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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- C05—FERTILISERS; MANUFACTURE THEREOF
- C05D—INORGANIC FERTILISERS NOT COVERED BY SUBCLASSES C05B, C05C; FERTILISERS PRODUCING CARBON DIOXIDE
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- C05—FERTILISERS; MANUFACTURE THEREOF
- C05G—MIXTURES OF FERTILISERS COVERED INDIVIDUALLY BY DIFFERENT SUBCLASSES OF CLASS C05; MIXTURES OF ONE OR MORE FERTILISERS WITH MATERIALS NOT HAVING A SPECIFIC FERTILISING ACTIVITY, e.g. PESTICIDES, SOIL-CONDITIONERS, WETTING AGENTS; FERTILISERS CHARACTERISED BY THEIR FORM
- C05G3/00—Mixtures of one or more fertilisers with additives not having a specially fertilising activity
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- 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
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- 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
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Abstract
The invention discloses a schizophyllum commune and a domestication and cultivation method and application thereof. The preservation number of the schizophyllum commune HMGIM-S140099 is CCTCC NO: M2016236. The schizophyllum commune HMGIM-S140099 is obtained through separation in Guangdong province, and the strain has very good medical and edible values and has a wide application prospect.
Description
Technical field：
The invention belongs to microorganism field, and in particular to one plant of schizophyllum commune and its domesticating cultivation method and application.
Background technology：
Schizophyllum commune (Schizophyllum commune Fr.) is commonly called as：Eight load bavins, spend in vain, smallpox bacterium, white ginseng bacterium, tree flower.
It is under the jurisdiction of mycota (Mycota), Eumycota (Eumycota), Basidiomycetes (Basidiomycetes), Agaricales
(Agaricales), schizophyllum commune section (Schizophyllaceae), Schizophyllum (Schizophyllum).
Schizophyllum sporophores head is little, in fan-shaped or palmate cracking, usual imbricate is storied, cluster is given birth to or all living creatures, is similar to little
Chrysanthemum.A length of 6～the 50mm of cap, wide 8～30mm, 1～3mm of thickness, cap superficial white, canescence, meat brown are close to yellowish-brown
Fine hair or coarse wool are draped over one's shoulders, with multiple rip-panels, tough meat to soft leather matter, edge is involute.Lamella white, canescence, light yellowish pink or band
Puce, Length discrepancy, pleat edge middle part lobe is into zanjon line.Bacterial context thickness about 1mm, white, keratin is flexible, tasteless, strong after doing to receive
Contracting, restores to the original state after water suction.Fructification stockless has short handle.Spore print is white or light yellowish pink, 5～7 × 2～3.5 μm of basidiospore,
Oval or sausage shape, smooth, colourless, non-starchy.
Distribution：Schizophyllum commune is distributed widely in all over the world, is distributed in major part provinces and cities of China, and microorganism resource is very rich
It is rich.
Habitat：It is scattered to all living creatures, it is often storied in rotten wood or bean curd stick
Drug effect：During schizophyllum sporophores children, matter is tender delicious, mild-natured with special strong fragrance, sweet, can nourish strong
Strengthen, strengthen the body resistance to consolidate the constitution, sedation, treatment neurasthenia, lassitude, dizzy tinnitus, abnormal sweating, are a kind of high-grade edible and medical fungis,
Its metabolite has preferable medical value.Schizophyllum sporophores or mycelium contain abundant immunity polysaccharide, protein,
Biologically active L-type amino acid, various essential amino acids and trace element etc., these materials are more to cardiovascular and liver, kidney etc. respectively
Kind of organ disease has a preventive and therapeutic effect, and the effects such as with anticancer, anti-aging, anti-inflammatory, with improving immunity of organisms, antibacterial
Various physiologically actives such as anti-inflammatory, radioresistance, with good edible and medical value.At present Schizophyllum commune Fr polysaccharides have been developed that into anti-
Antineoplastic agents, during the treatment for treating cervix cancer, cancer of the stomach and hepatitis B etc. is used for reference to chemicotherapy.
The content of the invention：
First purpose of the present invention be to provide can food and medicament dual-purpose, pick up from the schizophyllum commune of In Guangdong Province
(Schizophyllum commune) HMGIM-S140099, the bacterial strain is preserved in Chinese Typical Representative training on April 29th, 2016
Foster thing collection (abbreviation CCTCC, Wuhan City, Hubei Province Hongshan District Bayi Road, Wuhan University), preserving number is：CCTCC NO：M
2016236。
Second object of the present invention is to provide a kind of schizophyllum commune (Schizophyllum commune) HMGIM-
The domesticating cultivation method of S140099, it is characterised in that comprise the following steps：
Aseptically schizophyllum commune (Schizophyllum commune) HMGIM-S140099 parent species are inoculated into dress
In having the bag of pedigree seed culture medium, the shading culture under 25 DEG C of constant temperature, until mycelia eats full material and obtains final product original seed, original seed is in aseptic condition
Under be inoculated into the bag equipped with planting material culture medium, under 25 DEG C of constant temperature, relative air humidity be 60-70% lucifuge cultures, treat
Mycelia is covered with after planting material culture medium, carries out management of producing mushroom；
Described management of producing mushroom is to control temperature between 23 ± 1 DEG C, and between relative air humidity 60-80%, illumination is
100-200lux, control when carbon dioxide content has just gone out former base, in the sporophore growth phase, adjusts temperature 20- in more than 2000ppm
Between 23 DEG C, between relative air humidity 70-85%, illumination about 200-300lux, keep carbon dioxide content 1200ppm with
Under.
Described pedigree seed culture medium, in parts by weight, including Chinese sorghum 79%, millet 20%, CaCO31%, by its content
Each component is well mixed, is then added water, make water content be 55-60%, thus obtain pedigree seed culture medium.
Described planting material culture medium, in parts by weight, including cotton seed hulls 50%, wood chip 38%, the He of wheat bran 10%
CaCO32%, after by its content each component is well mixed, add water, make water content be 60%.
Third object of the present invention is to provide schizophyllum commune (Schizophyllum commune) HMGIM-S140099 and exists
Prepare the application in Schizophyllum commune Fr polysaccharides.
The present invention is from Guangdong Province isolated new strains schizophyllum commune (Schizophyllum commune) HMGIM-
There is S140099, the bacterial strain extraordinary medicine-food two-purpose to be worth, and have broad application prospects.
Schizophyllum commune (Schizophyllum commune) HMGIM-S140099, the bacterial strain is protected on April 29th, 2016
It is hidden in China typical culture collection center (abbreviation CCTCC, Wuhan City, Hubei Province Hongshan District Bayi Road, Wuhan University), preservation
Number it is：CCTCC NO：M 2016236.
Description of the drawings：
Fig. 1 is schizophyllum commune (Schizophyllum commune) HMGIM-S140099 and schizophyllum commune of the prior art
(Schizophyllum commune) Sc-1 and schizophyllum commune (Schizophyllum commune) Sc-2 antagonism figure between any two,
Wherein S140099, Sc-1 and Sc-2 represent respectively schizophyllum commune (Schizophyllum commune) HMGIM-S140099, split pleat
Bacterium (Schizophyllum commune) Sc-1 and schizophyllum commune (Schizophyllum commune) Sc-2.
Fig. 2 is the growth in the wild schizophyllum commune in White Cloud Mountain (Schizophyllum commune) HMGIM-S140099
Figure；
Fig. 3 is the growth figure of schizophyllum commune (Schizophyllum commune) HMGIM-S140099 after domesticating and cultivating.
Specific embodiment：
Following examples are that the present invention is further illustrated, rather than limitation of the present invention.
Embodiment 1：
First, schizophyllum commune (Schizophyllum commune) HMGIM-S140099 is obtained and identified：
Schizophyllum commune (Schizophyllum commune) HMGIM-S140099 of the present invention is isolated from Guangzhou, Guangdong
White Cloud Mountain broad-leaf forest falls on wood.
The morphological feature and ITS sequence of schizophyllum commune (Schizophyllum commune) HMGIM-S140099：
Schizophyllum sporophores all living creatures on withered tree, keratin, cap is fan-shaped or palmate, and a length of 7～15mm, wide 8～20mm are thick
2～3mm, surface canescence is had rug hair, and edge is involute.The light yellowish pink of lamella, Length discrepancy, pleat edge middle part lobe is into zanjon line.
Bacterial context white, about tough keratin, 1mm.Stem is as short as nothing.The light yellowish pink of spore print, spore is colourless under an optical microscope, transparent, spore
It is sub oval, 2.57～3.92 × 5.3～6.43 μm.
Schizophyllum commune (Schizophyllum is extracted using raw work Ezup pillar fungal genomic DNA extraction agent box
Commune) HMGIM-S140099 mycelium DNA, by primer of ITS1, ITS4 ITS-PCR is carried out, and after product sequencing alkali is obtained
Radix is the sequence of 636bp, and as shown in SEQ ID NO.1, Jing GenBank sequence Bs LAST compare analysis to its nucleotide sequence, with
The sequence similarity of the schizophyllum commune bacterial strain such as JX848644.1 for having delivered thereon and JF439509.1 up to 99%, it is determined that this point
From bacterial strain be Schizophyllum commun, be named as：Schizophyllum commune (Schizophyllum commune) HMGIM-
S140099。
2nd, antagonistic experiment
Plating medium (diameter 9cm) is made with common PDA culture medium, p H natures are connect respectively by triangle inocalation method
Enter schizophyllum commune (Schizophyllum commune) HMGIM-S140099, schizophyllum commune (Schizophyllum commune) Sc-
1 and schizophyllum commune (Schizophyllum commune) Sc-2 bacterial classification block (about 5mm square bacterium blocks), does respectively 3 repetitions, uses
After sealed membrane sealing, the lucifuge culture in 25 DEG C of constant incubators is mutually fully contacted to the mycelia of 3 kinds, sees whether exist
Antagonism line, and Taking Pictures recording.
As shown in Figure 1, schizophyllum commune (Schizophyllum commune) HMGIM-S140099 and of the prior art split
Gill fungus (Schizophyllum commune) Sc-1 and schizophyllum commune (Schizophyllum commune) Sc-2 are equal between any two
There is obvious antagonism line, cause mycelium that there is immiscible property because hereditary capacity is different, mycelium Antagonistic reaction be it is this not
Mutually melt one of macroscopic index of state, as a result show, schizophyllum commune (Schizophyllum commune) HMGIM-
S140099 and schizophyllum commune of the prior art (Schizophyllum commune) Sc-1 and schizophyllum commune (Schizophyllum
Commune) there is obvious hereditary difference between Sc-2, can determine whether schizophyllum commune (Schizophyllum commune)
HMGIM-S140099 and schizophyllum commune of the prior art (Schizophyllum commune) Sc-1 and schizophyllum commune
(Schizophyllum commune) Sc-2 is different strains.
3rd, domesticating and cultivating
1. culture medium
1) tissue is separated and mother culture media
Formula：Comprehensive PDA is (in parts by weight, including potato 20%+ glucose 2%+ agar 2%+ potassium dihydrogen phosphates
0.3%+ magnesium sulfate 0.15%+ vitamin B1s are micro, and solvent is water, i.e., its way is first potato to be cleaned into peeling, then is weighed
200g potatos are cut into small pieces, and add water well-done (boiling 20～30 minutes, can be poked by glass bar), with eight layers of gauze mistake
Filter, obtains filtrate, and 20g glucose, 20g agar are added in filtrate, and 3g potassium dihydrogen phosphates, 1.5g magnesium sulfate, vitamin B1 is micro, so
Afterwards 1L is supplied with water, sterilized.
2) pedigree seed culture medium
Formula：In parts by weight, including Chinese sorghum 79%, millet 20%, CaCO31%, each component is mixed by its content
Uniformly, then add water, make water content be 55-60%.
3) planting material culture medium
Tested by different formulations composition and comparision contents, finally determine optimum formula：In parts by weight, including cottonseed
Shell 50%, wood chip 38%, wheat bran 10%, CaCO32%, after mentioned component is well mixed, add water, make water content 60%.
2. cultivation technique parameter
1) cultural hypha
Separated by tissue under aseptic condition, obtain schizophyllum commune (Schizophyllum commune) HMGIM- after purification
S140099 parent species；Aseptically schizophyllum commune (Schizophyllum commune) HMGIM-S140099 parent species are inoculated with
To in the bag equipped with pedigree seed culture medium, the shading culture under 25 DEG C of constant temperature, until mycelia eats full material and obtains final product original seed.Original seed is aseptic
Under the conditions of be inoculated into the bag equipped with planting material culture medium, under 25 DEG C of constant temperature, relative air humidity be 60-70% lucifuges training
Support, after mycelia covers with planting material culture medium, carry out management of producing mushroom.The overall process is about needed 20-25 days.
2) Cultivation condition
Between 23 ± 1 DEG C, between relative air humidity 60-80%, illumination is about 100-200lux, dioxy to control temperature
Change when carbon content has just gone out former base and control in more than 2000ppm.Between sporophore growth phase, adjustment temperature 20-23 DEG C, air phase
To between humidity 70-85%, illumination about 200-300lux keeps carbon dioxide content in below 1200ppm.
3. fruiting situation
1) growth cycle：
Original seed is seeded into cultivating bag the long purseful of mycelia after 13-15 days, former base is gone out after 5-7 days, after 5-7 days fructification into
It is ripe to harvest, it is inoculated into the whole growth cycle of harvesting about 23-28 days from cultivating bag.
2) yield：Each bacterium bag is damp per between 45-60 gram of fruiting of tide, going out three, and overall biological conversion ratio is about in 70-85%
Between.
Fructification proterties：
4th, Thick many candies assay
1. the extraction of schizophyllum commune (Schizophyllum commune) HMGIM-S140099 fructification Thick many candies
Take schizophyllum commune after the cultivation of schizophyllum commune (Schizophyllum commune) HMGIM-S140099 strain domestications
Entity, after crushing, according to 1：30 solid-liquid ratio, extracts 2 hours at 100 DEG C, filters, with the anhydrous second of 4 times of volumes after concentration
Alcohol precipitates overnight, dialysis, obtains schizophyllum sporophores Thick many candies solution.
2. determination of polysaccharide
According in People's Republic of China's agricultural industry criteria (NY/T 1676-2008) edible mushroom Thick many candies containing measurement
Determine to schizophyllum commune (Schizophyllum commune) HMGIM-S140099 fructification Thick many candies content (Thick many candies/fructification
Dry weight × 100%) be measured.It is 5.17% to measure its polyoses content, it is documented that, Schizophyllum commune Fr polysaccharides (SPG) have anti-
Tumour, immunological regulation, antibacterial, antiviral etc. are acted on.Therefore, schizophyllum commune (Schizophyllum commune) HMGIM-
S140099 has preferably exploitation potential quality.
Sequence table
<110>Guangdong Microbes Inst（Microbiological analysis inspection center of Guangdong Province）
<120>One plant of schizophyllum commune and its domesticating cultivation method and application
<160> 1
<210> 1
<211> 636
<212> DNA
<213>Schizophyllum commune (Schizophyllum commune) HMGIM-S140099
<400> 1
TCCGTAGGTG AACCTGCGGA AGGATCATTA ACGAATCAAA CAAGTTCATC TTGTTCTGAT 60
CCTGTGCACC TCATGTAGTC CCAAAGCCTT CACGGGCGGC GGTTGACTAC GTCTACCTCA 120
CACCTTAAAG TATGTTAACG AATGTAATCA TGGTCTTGAC AGACCCTAAA AAGTTAATAC 180
AACTTTCGAC AACGGATCTC TTGGCTCTCG CATCGATGAA GAACGCAGCG AAATGCGATA 240
AGTAATGTGA ATTGCAGAAT TCAGTGAATC ATCGAATCTT TGAACGCACC TTGCGCCCTT 300
TGGTATTCCG AGGGGCATGC CTGTTTGAGT GTCATTAAAT ACCATCAACC CTCTTTTGAC 360
TTCGGTCTCG AGAGTGGCTT GGAAGTGGAG GTCTGCTGGA GCCTAACGGA GCCAGCTCCT 420
CTTAAATGTA TTAGCGGATT TCCCTTGCGG GATCGCGTCT CCGATGTGAT AATTTCTACG 480
TCGTTGACCA TCTCGGGGCT GACCTAGTCA GTTTCAATAG GAGTCTGCTT CTAACCGTCT 540
CTTGACCGAG ACTAGCGACT TGTGCGCTAA CTTTTGACTT GACCTCAAAT CAGGTAGGAC 600
TACCCGCTGA ACTTAAGCAT ATCAATAAGC GGAGGA 636
Claims (5)
1. schizophyllum commune (Schizophyllum commune) HMGIM-S140099, preserving number is：CCTCC NO：M 2016236.
2. a kind of domesticating cultivation method of schizophyllum commune (Schizophyllum commune) HMGIM-S140099, its feature exists
In comprising the following steps：
Aseptically by schizophyllum commune (Schizophyllum commune) the HMGIM-S140099 parent species in claim 1
In being inoculated into the bag equipped with pedigree seed culture medium, the shading culture under 25 DEG C of constant temperature, until mycelia eats full material and obtains final product original seed, original seed exists
During the bag equipped with planting material culture medium is inoculated under aseptic condition, under 25 DEG C of constant temperature, relative air humidity is 60-70% lucifuges
Culture, after mycelia covers with planting material culture medium, carries out management of producing mushroom；
Described management of producing mushroom is to control temperature between 23 ± 1 DEG C, and between relative air humidity 60-80%, illumination is 100-
200lux, control when carbon dioxide content has just gone out former base, in the sporophore growth phase, adjusts temperature 20-23 DEG C in more than 2000ppm
Between, between relative air humidity 70-85%, illumination about 200-300lux keeps carbon dioxide content in below 1200ppm.
3. preparation method according to claim 2, it is characterised in that described pedigree seed culture medium, in parts by weight, bag
Include Chinese sorghum 79%, millet 20%, CaCO31%, each component is well mixed by its content, then add water, make the water content be
55-60%, thus obtains pedigree seed culture medium.
4. preparation method according to claim 2, it is characterised in that described planting material culture medium, in parts by weight,
Including cotton seed hulls 50%, wood chip 38%, wheat bran 10% and CaCO32%, after by its content each component is well mixed, then add
Water, makes water content be 60%.
5. schizophyllum commune (Schizophyllum commune) HMGIM-S140099 described in claim 1 is to prepare schizophyllum commune more
Application in sugar.
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Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107736190A (en) * | 2017-11-13 | 2018-02-27 | 云南菌视界生物科技有限公司 | A kind of waste material after fruiting using schizophyllum commune cultivates the method that high warm lung shape is picked up the ears |
| CN107828664A (en) * | 2017-10-25 | 2018-03-23 | 湖南省农业生物技术研究中心 | Schizophyllum commune XT 1 and its cultural method and application |
| CN109156266A (en) * | 2018-08-03 | 2019-01-08 | 西北师范大学 | A kind of method schizophyllum commune cultivation culture medium and cultivate schizophyllum commune using it |
| CN109247189A (en) * | 2018-09-29 | 2019-01-22 | 云南菌视界生物科技有限公司 | A kind of white ginseng bacterial strain and its white ginseng low cost cultural method |
| CN113179853A (en) * | 2021-04-28 | 2021-07-30 | 马布平 | White ginseng strain, cultivation field and white ginseng |
| CN114214209A (en) * | 2021-12-27 | 2022-03-22 | 宁波希诺亚海洋生物科技有限公司 | Schizophyllum commune variety capable of producing lactase and application thereof |
| CN114391421A (en) * | 2022-01-06 | 2022-04-26 | 毕节市农投菌业科技有限责任公司 | Cultivation method for continuously fruiting white ginseng fungi |
| CN116590156A (en) * | 2023-06-20 | 2023-08-15 | 南京高新工大生物技术研究院有限公司 | Schizophyllum commune strain with high protein yield and breeding method thereof |
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Cited By (12)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107828664A (en) * | 2017-10-25 | 2018-03-23 | 湖南省农业生物技术研究中心 | Schizophyllum commune XT 1 and its cultural method and application |
| CN107828664B (en) * | 2017-10-25 | 2020-10-23 | 湖南省农业生物技术研究中心 | Schizophyllum commune XT-1 and cultivation method and application thereof |
| CN107736190A (en) * | 2017-11-13 | 2018-02-27 | 云南菌视界生物科技有限公司 | A kind of waste material after fruiting using schizophyllum commune cultivates the method that high warm lung shape is picked up the ears |
| CN109156266A (en) * | 2018-08-03 | 2019-01-08 | 西北师范大学 | A kind of method schizophyllum commune cultivation culture medium and cultivate schizophyllum commune using it |
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| CN116590156B (en) * | 2023-06-20 | 2024-01-30 | 南京高新工大生物技术研究院有限公司 | Schizophyllum commune strain with high protein yield and breeding method thereof |
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