CN105385605A - Poria cocos fast-growing strains and preparation method thereof - Google Patents
Poria cocos fast-growing strains and preparation method thereof Download PDFInfo
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- CN105385605A CN105385605A CN201510677998.3A CN201510677998A CN105385605A CN 105385605 A CN105385605 A CN 105385605A CN 201510677998 A CN201510677998 A CN 201510677998A CN 105385605 A CN105385605 A CN 105385605A
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- poria cocos
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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
Abstract
The invention relates to poria cocos fast-growing strains in the field of fungi and a preparation method thereof, and belongs to strain production techniques in poria cocos industry development. The poria cocos fast-growing strains are characterized in that ground-stage seeds are transferred by a poria cocos test tube, fluid suspension culture is performed, and third-stage strains are inoculated and cultured on raw wood sticks; the third-stage strain production period is only 4-8 days. According to the poria cocos fast-growing strains and the technique thereof, an original production method of the poria cocos strains is changed, the defects of the original production method are overcome, and the poria cocos industry development is facilitated.
Description
Technical field
The present invention relates to Poria cocos bacterial classification production technical field, be specifically related to a kind of preparation method of Poria cocos fast-growing bacterial classification.
Background technology
Poria cocos (formal name used at school: Poriacocos (Schw.) Wolf.) has another name called: Fu Tu, Fu spirit, and Fu Ling, Fu Ling, volt Tu, pine fat, deep red morning lies prostrate tire, Yun Ling, Fu rabbit, loose potato, pine potato, Song Ling.Medicinal part is the sclerotium of On Polyporaceae Poria cocos.There is promoting diuresis to eliminate damp pathogen, invigorating the spleen, the effect of calming the nerves.Can also be made into powdery be used for Ms hobby cleawhite pack.Distributed areas: Jilin, Anhui, Zhejiang, Fujian, Henan, Hubei, Guangxi, Sichuan, Guizhou, Yunnan.Ancients claim Poria cocos to be " when four god medicine ", because its effect widely, regardless of the four seasons, by it and various compatibility of drugs, no matter cold, temperature, wind, wet all diseases, can play its unique effects.Poria cocos taste is sweet, light, property is put down, and is used as medicine and has the function of promoting diuresis to eliminate damp pathogen, beneficial spleen and stomach, tranquillizing by calming the heart.Modern medicine study: Poria cocos energy enhancing body immunologic function, Pachymose has significantly antitumor and protect the liver dirty effect.
For good medical treatment and the health value of Poria cocos, since ancient times, our ancestors have carried out many Application and Development to it, as pharmaceutical applications, its many and other drug compatibility, numerous medicine, food research institute develop it, the product being used as medicine in the market has guizhi fuling pill, Poria cocos radix polygonati officinalis sheet, Poria cocos yam slice, the former powder of Pachymose, oral liquor of poria polysaccharide, Pachymose compound preparation etc., has cake, tuckahoe cream, ant tuckahoe wine etc. as protective foods.Its market outlook are very wide.
The major ingredient of Poria cocos is β-pachyman (also known as " Pachymose "), accounts for 93% of dry weight.Pachymose is immunostimulant, has and strengthens immunity, adjuvant chemotherapy, anti-curing oncoma, the effect such as antitumor, be widely used as PTS,
Poria cocos is the warm fungi of happiness, and suitable China, South East Asia, America, Oceania etc. originated in has the mountain area of pine tree local.China is Poria cocos main product state, accounts for 70% of world's Poria cocos ultimate production.In recent years, China's year Poria cocos product demand amount is more than 20,000 tons, and annual export volume is about 7000 tons, and market demand presents cumulative year after year trend.But, both at home and abroad about the research of Poria cocos new mushroom-seed culturing breeding technique is still inadequate; And bacterial classification is through successive propagation for many years, occurs natural variation, deterioration of strains, bacterial strain vitality declines, and Siberian cocklebur body is little, and biological efficiency declines, and there is plantation success ratio, Poria cocos output, Poria cocos quality are extremely low, and resource scarcity and the problem such as production cost is high, economic benefit is low.Therefore, the Poria cocos new mushroom-seed culturing that seed selection quality is more excellent, improves Yield and quality and the industrial benefit of Poria cocos plant husbandry, people current in the urgent need to address be technical problem.
Summary of the invention
In order to solve the problem, embodiments provide a kind of Poria cocos fast-growing bacterial classification and preparation method thereof on the one hand, bacterium is cultivated, solid-state inoculation cultivation to adopt liquid to send out, anti-hybrid ability is strong, wide adaptability, tuckahoe production rate is high, effectively shortens the growth cycle of Poria cocos, decrease the pollution of miscellaneous bacteria and the harm of termite, there is the advantages such as high-quality, high yield, high resistance;
In another aspect the preferred embodiments of the invention, the major traits feature of Poria cocos fast-growing bacterial classification comprises: 1) bacterial classification is with short production cycle, and mycelium is sturdy, white, and clamp connexion body is obvious; In planting process, inoculation is convenient, and mycelia passes to be drawn soon; 2) tie Siberian cocklebur early, output is high, and Siberian cocklebur body is evenly solid, meat white, and Siberian cocklebur skin is thin, in Zhu's look; 3) there is stabilization characteristics of genetics, anti-hybrid ability is strong, the advantages such as quality is good;
The object of the present invention is to provide a kind of preparation method of Poria cocos fast-growing bacterial classification, comprise the steps:
1) female cultivation of planting: select its Siberian cocklebur body evenly solid, look red, Siberian cocklebur skin is thin, give off a strong fragrance, the measured wild Poria cocos of matter, tissue culture is carried out under 24-28 DEG C of condition, temperature 24 DEG C-26 DEG C, during atmospheric moisture 70%-80%, Poria cocos epidermis is taken out with aseptic cutter, get a small pieces entity, isolate pyrenomycetes filament is cultivated, and obtains female kind; , wherein, the moisture content in medium of described tissue culture is 55-60%, pH value 5-6, and composition comprises the carbon source of suitable Poria cocos growth, and described carbon source is selected from sucrose, glucose, nitrogenous source are selected from peptone, amino acid;
2) mycelial purification, rejuvenation: mother planted and be inoculated in test-tube culture medium, be placed in thermostat container, cultivates 7-10 days, covers with substratum to mycelia for 26-28 DEG C, selects the most the superior of its upgrowth situation and carries out rejuvenation cultivation,
3) described mycelial cultivation, purification, rejuvenation culture medium consist of potato 200g, agar 20g, glucose 15g, brown sugar 15g, potassium primary phosphate 1.5g, magnesium sulfate 0.75g, VB11 g, peptone 5g, water 1000mi(pine sawdust juice).Its preparation method: comprise the steps: that (1) is by peeling potatoes, cleans clean, thinly slices, put into pot and boil, boil thoroughly well cooked but not mushy degree, screen filtration, and obtained potato juice, regulates its pH value 5.5-6.5; (2) in obtained potato juice, add other composition in substratum composition, after heating for dissolving, after being distributed into test tube, high-temperature sterilization 30 minutes under 0.15 kilogram of pressure, takes out, and cooling, is made into slant medium.Aseptically access Poria cocos mother to plant; The mother that must purify plants,
4) selection of Poria cocos fast-growing bacterial classification: according to the seed selection ultimate principle of biological gene heredity, induce variation and new variety, select Poria cocos fast-growing bacterial classification pointedly; Wherein, the substratum of described mycelial cultivation, purification, rejuvenation consists of, potato 200g, agar 20g, glucose 15g, brown sugar 15g, potassium primary phosphate 1.5g, magnesium sulfate 0.75g, VB11 g, peptone 5g, water 1000mi(pine sawdust juice).
5) in the preferred technical solution of the present invention, described mycelial cultivation, purification, the consisting of of rejuvenation culture medium, potato 200g, agar 20g, glucose 15g, brown sugar 15g, potassium primary phosphate 1.5g, magnesium sulfate 0.75g, VB11 g,, peptone 5g, water 1000mi(pine sawdust juice).Its preparation method: comprise the steps: that (1) is by peeling potatoes, cleans clean, thinly slices, put into pot and boil, boil thoroughly well cooked but not mushy degree, screen filtration, and obtained potato juice, regulates its pH value 5.5-6.5; (2) in obtained potato juice, add other composition in substratum composition, after heating for dissolving, after being distributed into test tube, high-temperature sterilization 30 minutes under 0.15 kilogram of pressure, takes out, and cooling, is made into slant medium.Aseptically access Poria cocos mother to plant;
6) the invention reside in the preparation of the I and II liquid providing a kind of Poria cocos fast-growing bacterial classification, it is characterized in that, described method comprises the steps:
(1) one-level is cultivated: access described mother in liquid medium within and plant, cultivate 7 days, as primary seed solution;
(2) secondary is cultivated: described first-generation seed liquor pressed in the inoculum size access liquid nutrient medium of culture volume 10%-15%, cultivate 7 days, as secondary seed solution;
7) the invention reside in the preparation of the three grades of cultivars providing a kind of Poria cocos fast-growing bacterial classification, it is characterized in that, described method comprises the steps:
Select loose section wood that is fresh, dry, inclusion-free pollution, its standard: long 15-18mm, diameter 6-8mm, after the sun is exposed to the sun drying, requires stringent sterilization, sterilization, in sterilisable chamber aseptic inoculation secondary liquid kind; Inoculum size is 30-40%, and cultivation temperature is 28-30 DEG C, and the cultivation time is 4-8 days, after mycelia is covered with, obtains Poria cocos fast-growing bacterial classification;
8) a kind of Poria cocos fast-growing bacterial classification of the present invention is characterized in that:
(1) bacterial classification is with short production cycle, and mycelium is sturdy, white, and clamp connexion body is obvious; In planting process, inoculation is convenient, and mycelia passes to be drawn soon;
(2) tie Siberian cocklebur early, output is high, and Siberian cocklebur body is evenly solid, meat white, and Siberian cocklebur skin is thin, in Zhu's look;
(3) have stabilization characteristics of genetics, anti-miscellaneous bacteria ability is strong, quality is good, adaptable, the advantage such as all can to plant the four seasons;
(4) content of the effective constituents such as the Pachymose in Poria cocos is significantly improved.
In the embodiment of the present invention, made be improved to liquid culture by traditional solid, the three grades of wood chip kind section of being improved to wood are planted, and strengthen the stable of bacterial classification genetic process, control the degeneration in the stage of breeding.Can cultivate in the four seasons, have adjusted the labor force of busy farming season significantly, have adjusted the high temperature time of weather; Effectively enhance the quality of production of Poria cocos, improve the output of Poria cocos plantation, Poria cocos industry is reached for the purpose of high yield and high quality.
Embodiment
For making the object, technical solutions and advantages of the present invention clearly, below embodiment of the present invention is described further in detail, but this and do not mean that any limitation of the invention.
1) the one-level mother making of planting: select Siberian cocklebur body evenly solid, look red, Siberian cocklebur skin is thin, give off a strong fragrance, the measured wild Poria cocos of matter, tissue culture is carried out under 24-28 DEG C of condition, temperature 24 DEG C-26 DEG C, during atmospheric moisture 70%-80%, Poria cocos epidermis is taken out with aseptic cutter, get a small pieces entity, separation sclerotium and mycelia are cultivated, and obtain female kind;
2) making of mother culture media: potato 200g, agar 20g, glucose 15g, brown sugar 15g, potassium primary phosphate 1.5g, magnesium sulfate 0.75g, VB11 g, peptone 5g, water 1000mi(pine sawdust juice).Its preparation method: comprise the steps:
(1) by peeling potatoes, clean clean, thinly slice, put into pot and boil, boil thoroughly well cooked but not mushy degree, screen filtration, obtained potato juice, regulates its pH value 5.5-6.5;
(2) in obtained potato juice, add other composition in substratum composition, after heating for dissolving, after being distributed into test tube, high-temperature sterilization 30 minutes under 0.15 kilogram of pressure, takes out, and cooling, is made into slant medium.
The making of liquid spawn
1) one-level is cultivated: in one-level nutrient solution, access Poria cocos mother plant, 96-100 hour is cultivated under 28-30 DEG C of condition, obtain primary seed solution, the formula of described first cell culture medium with weight is: potato 20-25%, glucose 2-2.5%, potassium primary phosphate 0.2-0.3%, magnesium sulfate 0.15-0.2%, surplus are water, VB11-2/L;
2) secondary is cultivated: be linked in secondary medium by described primary seed solution by the inoculum size of culture volume 10-15%, 96-100 hour is cultivated under 28-30 DEG C of condition, obtain described Poria cocos seed liquor, the formula of described secondary medium with weight is: glucose 2-2.5%, brown sugar 2.5-3%, potassium primary phosphate 0.2-0.3%, magnesium sulfate 0.15-0.2%, corn steep liquor 2.5-3%, yeast extract paste 2-2.5%, surplus are water, VB11-2/L.
The making of cultivar: select fresh, dry loose section wood, its standard: long 15-18mm, diameter 6-8mm, after sterilizing, sterilization, after the sun is exposed to the sun drying, requires strict dry, clean, inclusion-free, in sterilisable chamber aseptic inoculation secondary liquid kind; Inoculum size is 30-40%, and cultivation temperature is 28-30 DEG C, and the cultivation time is 4-8 days, and mycelia covers with charge level, pack; Obtain Poria cocos fast-growing bacterial classification;
The cultivation of Poria cocos fast-growing bacterial classification and the common bacterial classification of Poria cocos contrasts, and difference is: fast-growing bacterial classification has sends out the advantages such as bacterium is fast, the time is short, strong stress resistance, surviving rate are high; It contrasts situation as shown in Table 1:
| Contrast title | Adaptive temperature DEG C | The Mother culture time (my god) | Liquid strain incubation time (my god) | Culture spawn incubation time (my god) | The preservation time (natural temperature) (my god) | Seed yield rate % |
| Fast-growing bacterial classification | 20-35 | 6-10 | 4-6 | 4-8 | 10-60 | 85-100 |
| Common bacterial classification | 25-30 | 6-10 | 25-35 | 25-35 | 10-20 | 75-85 |
The foregoing is only preferred embodiment of the present invention, not in order to limit the present invention, within the spirit and principles in the present invention all, any amendment done, equivalent replacement, improvement etc., all should be included within protection scope of the present invention.
Claims (8)
1. Poria cocos fast-growing bacterial classification and preparation method thereof, is characterized in that, the wild seed selection of test tube kind in Poria cocos bacterial classification production technique, first order seed suspension culture, and secondary seed liquid is cultivated, and three grades of seed raw material wooden sticks are sent out bacterium and cultivated.
2. the preparation method of Poria cocos fast-growing bacterial classification according to claim 1, comprises the steps:
One-level is cultivated: in first cell culture medium, access Poria cocos mother plant, 96-100 hour is cultivated under 28-30 DEG C of condition, obtain primary seed solution, the formula of described first cell culture medium is counted with the content of weight percent: potato 20-25%, glucose 2-2.5%, potassium primary phosphate 0.2-0.3%, magnesium sulfate 0.15-0.2%, surplus are water, VB11-2/L;
Secondary is cultivated: be linked in secondary medium by described primary seed solution by the inoculum size of culture volume 10-15%, 96-100 hour is cultivated under 28-30 DEG C of condition, obtain described Poria cocos seed liquor, the formula of described secondary medium is counted with the content of weight percent: glucose 2-2.5%, brown sugar 2.5-3%, potassium primary phosphate 0.2-0.3%, magnesium sulfate 0.15-0.2%, corn steep liquor 2.5-3%, yeast extract paste 2-2.5%, surplus are water, VB11-2/L.
3. the preparation method of Poria cocos fast-growing bacterial classification according to claim 2, is characterized in that, the bacterium condition of sending out that step 302 secondary liquid kind is linked into three grades of solid kinds is: aseptic inoculation secondary liquid kind; Inoculum size is 30-40%, and cultivation temperature is 28-30 DEG C, and the cultivation time is 4-6 days, and mycelia covers with charge level.
4. the preparation method of Poria cocos fast-growing bacterial classification according to claim 2, is characterized in that:
Select fresh, dry loose section wood, its standard: long 15-18mm, diameter 6-8mm, after the sun is exposed to the sun drying, requires stringent sterilization, sterilization, in sterilisable chamber aseptic inoculation secondary liquid kind; Be 30-40% by step 303 inoculum size, cultivation temperature is 28-30 DEG C, and the cultivation time is 4-6 days, and mycelia covers with charge level, obtains Poria cocos fast-growing bacterial classification.
5. the preparation method of Poria cocos fast-growing bacterial classification according to claim 2, comprises the steps:
The bacterial strain of Poria cocos fast-growing bacterial classification is selected and is cultivated: according to ultimate principle and the territorial environment requirement of biological heredity, variation, seed selection new variety, select the bacterial strain of Poria cocos fast-growing bacterial classification pointedly, therefrom select the improved seeds of Poria cocos fast-growing bacterial classification; Select Siberian cocklebur body evenly solid, look red, Siberian cocklebur skin is thin, give off a strong fragrance, the measured wild Poria cocos of matter, tissue culture is carried out under 24-28 DEG C of condition, temperature 24 DEG C-26 DEG C, during atmospheric moisture 70%-80%, Poria cocos epidermis is taken out with aseptic cutter, get a small pieces entity, separation sclerotium and mycelia are cultivated, and obtain female kind.
6. selection according to claim 3, is characterized in that;
Described mycelial cultivation, purification, rejuvenation culture medium consist of potato 200g, agar 20g, glucose 15g, brown sugar 15g, potassium primary phosphate 1.5g, magnesium sulfate 0.75g, VB11 g, peptone 5g, water 1000mi(pine sawdust juice), its preparation method: comprise the steps: that (1) is by peeling potatoes, cleans clean, thinly slice, put into pot to boil, boil thoroughly well cooked but not mushy degree, screen filtration, obtained potato juice, regulates its pH value 5.5-6.5; (2) in obtained potato juice, other composition in substratum composition is added, after heating for dissolving, after being distributed into test tube, high-temperature sterilization 30 minutes under 0.15 kilogram of pressure, takes out, cooling, be made into slant medium, aseptically access Poria cocos mother and plant; The mother that must purify plants.
7. Poria cocos fast-growing spawn culture method according to claim 4, is characterized in that;
Gained Poria cocos fast-growing bacterial classification is purified and is planted, be made into liquid spawn, comprise the steps: that in nutrient solution, access Poria cocos mother plants, 96-100 hour is cultivated under 28-30 DEG C of condition, obtain primary seed solution, the formula of described first cell culture medium with weight is: potato 200g, agar 20g, glucose 20g, potassium primary phosphate 1.5g, magnesium sulfate 0.75g, vitaminB10 .75g, peptone 5g, water 1000mi, obtain liquid strain.
8. Poria cocos fast-growing spawn culture method gained Poria cocos fast-growing strain liquid kind according to claim 5, is characterized in that, described method, comprises the steps:
(1) one-level is cultivated: in liquid medium, access described mother plant, cultivate 7 days, as primary seed solution;
(2) secondary is cultivated: described first-generation seed liquor pressed in the inoculum size access liquid nutrient medium of culture volume 10%-15%, cultivate 7 days, as secondary seed solution;
Poria cocos fast-growing spawn culture method gained Poria cocos fast-growing bacterial classification according to claim 6 three grades of kinds, is characterized in that described method, comprise the steps:
Select loose section wood that is fresh, dry, inclusion-free pollution, its standard: long 15-18mm, diameter 6-8mm, after the sun is exposed to the sun drying, requires stringent sterilization, sterilization, in sterilisable chamber aseptic inoculation secondary liquid kind; Inoculum size is 30-40%, and cultivation temperature is 28-30 DEG C, and the cultivation time is 4-8 days, after mycelia is covered with, obtains Poria cocos fast-growing bacterial classification.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106398963A (en) * | 2016-10-25 | 2017-02-15 | 柳杨丽 | Poria cocos solid fermented wine and preparation method thereof |
| CN106962022A (en) * | 2017-05-10 | 2017-07-21 | 黎平绿华农林发展有限公司 | A kind of loose waste material adds the Poria cocos breeding method of corn |
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2015
- 2015-10-20 CN CN201510677998.3A patent/CN105385605A/en active Pending
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106398963A (en) * | 2016-10-25 | 2017-02-15 | 柳杨丽 | Poria cocos solid fermented wine and preparation method thereof |
| CN106962022A (en) * | 2017-05-10 | 2017-07-21 | 黎平绿华农林发展有限公司 | A kind of loose waste material adds the Poria cocos breeding method of corn |
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