CN101240241B - Method for preserving stropharia rugoso-annulata strain - Google Patents
Method for preserving stropharia rugoso-annulata strain Download PDFInfo
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- CN101240241B CN101240241B CN2008100176944A CN200810017694A CN101240241B CN 101240241 B CN101240241 B CN 101240241B CN 2008100176944 A CN2008100176944 A CN 2008100176944A CN 200810017694 A CN200810017694 A CN 200810017694A CN 101240241 B CN101240241 B CN 101240241B
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Abstract
The invention discloses a preserving method of large ball-cover mushroom strain comprising a) configuring directing domestication plating medium, b) preparing non-freezing, non-negative pressure mycelium, c) freezing and drying the mycelium in vacuum: putting the ampoule pipe into refrigerating drier to freeze to -40 DEG C at the speed of 2 DEG C per minute, pre-freezing in constant temperature for 1.5 hours and then opening the vacuum pump, after the vacuum degree achieving 22.5 Pa within 15 minutes, the temperature of the isolation plate rising 5 DEG C and the vacuum degree reducing 5Pa per30 minutes until the temperature of the isolation plate rising to 24 DEG C and the vacuum degree reducing to 0.8 to.
Description
Technical field
The present invention relates to a kind of edible fungus species method for preserving, is the culture collection process of Stropharia rugoso-annulata specifically.
Background technology
Bacterial classification is as a kind of important Biological resources, in order to keep its good character constant, should in time carry out preservation to it, the edible fungus species method for preserving mainly contains liquid nitrogen cryogenics preserving process, the regular transfer methods in inclined-plane, whiteruss preserving process, sand tube preservation method and natural matrix preserving process etc. at present, wherein the liquid nitrogen cryogenics preserving process is comparatively advanced at present a kind of method, but expense is higher, and external preservation mechanism adopts this method more; In China, produce at present or research in institute's edible fungus species still mainly adopt test tube or bottle preserving process, main drawback has: the shelf time is shorter, bacterial classification stability is bad, easily morphs or degenerates, the cost height carries shortcomings such as inconvenience.
Existing edible mushroom mycelium live dry strain and production method thereof, freeze-drying obtains edible mushrooms dried mycelium bacterial classification product alive as the material use Vacuum Freezing ﹠ Drying Technology with mycelium, but practice shows that mycelium is unsuitable for freeze-drying.
Summary of the invention
The object of the invention provides a kind of method for preserving stropharia rugoso-annulata strain,
A), configuration guiding domestication plate culture medium: its compositing formula: wheat extracting solution 20g, yeast extract 1g, sucrose 0.75g, lactose 1g, potassium primary phosphate 2g, sal epsom 1g, sodium-chlor 0.3g, ironic citrate 0.2g, agar 1.2g and distilled water 92.55g, pH6.8, make aseptic plate, when treating culture medium solidifying, lay one deck in media surface and lay one deck porous paper in media surface.
B), anti-ly freeze, the mycelium preparation of anti-negative pressure: when treating that then substratum is cooled to 20～25 ℃; with the Stropharia rugoso-annulata bacterial classification inoculation to carrier; in 25 ℃ of sterile culture casees, cultivated 144 hours; under aseptic condition, carrier is taken off in the lump together with bacterial classification then; and be trimmed to corresponding size according to ampoul tube (bottle) bottom size and be positioned over bottom the ampoul tube (bottle); add protective material 0.1～0.2mL, its prescription is as follows: skimmed milk 18g, glucose 1g, N.F,USP MANNITOL 2g, Mierocrystalline cellulose 1g and distilled water 100ml.
C), mycelium vacuum lyophilization: ampoul tube is put into freeze drier, cooling rate is 2 ℃ of per minutes, is cooled to-40 ℃, constant temperature pre-freeze 1.5 hours, open vacuum pump then, after vacuum tightness reaches 22.5 handkerchiefs in 15 minutes, raise 5 ℃ with per 30 minutes baffle temperature, vacuum tightness reduces by 5 handkerchiefs, treat that baffle temperature is increased to 24 ℃, vacuum tightness drops to 0.8～1.3 handkerchief, and when sample temperature was increased to 24 ℃, vacuum lyophilization was finished.
The present invention, prepares freeze pipe then and carries out preservation to improve performances such as its low temperature frost resistance, anti-negative pressure and rehydration surviving rate by to Stropharia rugoso-annulata mycelium channeling conduct domestication.
That the present invention has is applied widely, preservation term long, the surviving rate advantages of higher.
Embodiment
The following examples can make those skilled in the art more fully understand the present invention, but do not limit the present invention in any way.
Embodiment 1:
At first plate culture medium is tamed in the configuration guiding; its compositing formula: wheat extracting solution 20g; yeast extract 1g; sucrose 0.75g; lactose 1g; potassium primary phosphate 2g; sal epsom 1g; sodium-chlor 0.3g; ironic citrate 0.2g; agar 1.2g; distilled water 92.55g; pH6.8; make aseptic plate; when treating that substratum slightly solidifies; lay one deck in media surface and lay one deck porous paper in media surface; when treating that then substratum is cooled to 20～25 ℃; with the Stropharia rugoso-annulata bacterial classification inoculation to carrier; in 25 ℃ of sterile culture casees, cultivated 144 hours; under aseptic condition, carrier is taken off in the lump together with bacterial classification then; and be trimmed to corresponding size according to ampoul tube (bottle) bottom size and be positioned over bottom the ampoul tube (bottle); add protective material 0.1～0.2mL; its prescription is as follows: skimmed milk 18g; glucose 1g; N.F,USP MANNITOL 2g; Mierocrystalline cellulose 1g; distilled water 100ml; ampoul tube is put into freeze drier; cooling rate is 2 ℃ of per minutes; be cooled to-40 ℃; constant temperature pre-freeze 1.5 hours; open vacuum pump then; after vacuum tightness reaches 22.5 handkerchiefs in 15 minutes; raise 5 ℃ with per 30 minutes baffle temperature; vacuum tightness reduces by 5 handkerchiefs; treat that baffle temperature is increased to 24 ℃; vacuum tightness drops to 0.8～1.3 handkerchief; when sample temperature was increased to 24 ℃, vacuum lyophilization was finished.The rehydration experimental result shows that the bacterial classification survival rate is more than 75%.
Claims (1)
1. method for preserving stropharia rugoso-annulata strain is characterized in that: it comprises the following steps,
A), configuration guiding domestication plate culture medium: the substratum compositing formula is wheat extracting solution 20g, yeast extract 1g, sucrose 0.75g, lactose 1g, potassium primary phosphate 2g, sal epsom 1g, sodium-chlor 0.3g, ironic citrate 0.2g, agar 1.2g and distilled water 92.55g, pH6.8, make aseptic plate, when treating culture medium solidifying, lay one deck porous paper in media surface;
B), anti-ly freeze, the mycelium preparation of anti-negative pressure: when treating that substratum is cooled to 20～25 ℃, with the Stropharia rugoso-annulata bacterial classification inoculation to carrier, in 25 ℃ of sterile culture casees, cultivated 144 hours, under aseptic condition, carrier is taken off in the lump together with bacterial classification then, and be trimmed to corresponding size according to ampoul tube bottom size and be positioned over the ampoul tube bottom, add protective material 0.1～0.2mL, the protective material prescription is skimmed milk 18g, glucose 1g, N.F,USP MANNITOL 2g, Mierocrystalline cellulose 1g and distilled water 100ml;
C), mycelium vacuum lyophilization: ampoul tube is put into freeze drier, cooling rate is 2 ℃ of per minutes, is cooled to-40 ℃, constant temperature pre-freeze 1.5 hours, open vacuum pump then, after vacuum tightness reaches 22.5 handkerchiefs in 15 minutes, raise 5 ℃ with per 30 minutes baffle temperature, vacuum tightness reduces by 5 handkerchiefs, treat that baffle temperature is increased to 24 ℃, vacuum tightness drops to 0.8～1.3 handkerchief, and when sample temperature was increased to 24 ℃, vacuum lyophilization was finished.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2008100176944A CN101240241B (en) | 2008-03-04 | 2008-03-04 | Method for preserving stropharia rugoso-annulata strain |
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| Application Number | Priority Date | Filing Date | Title |
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| CN2008100176944A CN101240241B (en) | 2008-03-04 | 2008-03-04 | Method for preserving stropharia rugoso-annulata strain |
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| Publication Number | Publication Date |
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| CN101240241A CN101240241A (en) | 2008-08-13 |
| CN101240241B true CN101240241B (en) | 2010-06-09 |
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| CN2008100176944A Expired - Fee Related CN101240241B (en) | 2008-03-04 | 2008-03-04 | Method for preserving stropharia rugoso-annulata strain |
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Families Citing this family (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102498926A (en) * | 2011-10-08 | 2012-06-20 | 金乡县益佳康生物科技有限责任公司 | Production method of liquid lyophilized strain of agaric |
| CN102586114A (en) * | 2012-02-21 | 2012-07-18 | 常熟市方塔涂料化工有限公司 | Preservation method for mushroom strain |
| CN111742785B (en) * | 2020-07-07 | 2022-05-06 | 辽东学院 | Long-term preservation method of cordyceps militaris strain with high cordycepin yield |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1164572A (en) * | 1997-04-24 | 1997-11-12 | 万兰英 | Edible mushroom mycelium live dry strain and its producing method |
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Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1164572A (en) * | 1997-04-24 | 1997-11-12 | 万兰英 | Edible mushroom mycelium live dry strain and its producing method |
Non-Patent Citations (2)
| Title |
|---|
| 闫培生等.大球盖菇菌丝生长的营养需求及环境条件.食用菌学报8 1.2001,8(1),第5页1. 2. 2，1. 3. 1，1. 3. 2，1. 3. 3，1. 3. 4段，第6页1.3. 7段，第7页2.4段. |
| 闫培生等.大球盖菇菌丝生长的营养需求及环境条件.食用菌学报8 1.2001,8(1),第5页1. 2. 2,1. 3. 1,1. 3. 2,1. 3. 3,1. 3. 4段,第6页1.3. 7段,第7页2.4段. * |
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