CN101701179B - Device and method for purifying mycelial fungi polluted by bacteria - Google Patents
Device and method for purifying mycelial fungi polluted by bacteria Download PDFInfo
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- CN101701179B CN101701179B CN 200910235086 CN200910235086A CN101701179B CN 101701179 B CN101701179 B CN 101701179B CN 200910235086 CN200910235086 CN 200910235086 CN 200910235086 A CN200910235086 A CN 200910235086A CN 101701179 B CN101701179 B CN 101701179B
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Abstract
The invention discloses a device and a method for purifying mycelial fungi polluted by bacteria. The device is formed by packing a fungus purification layer and a biologic purifying column with a seal film and a purification cup. The method comprises the steps of making the biologic purifying column after adding fungus nutrient solution in a porous material, making a purified fungus collecting layer based on a fungus solid nutrient medium, growing the fungi polluted by bacteria on the biologic purifying column, detaining the fungus on the biologic purifying column by utilizing the selectivity using of mycelial fungi and bacteria to the nutriment of the nutrient medium and the difference of the growing characteristics in materials, and then outgrowing the biologic purifying column on fungi and growing in the purified fungus collecting layer to conveniently and efficiently purify the fungi polluted by bacteria.
Description
Technical field:
The present invention relates to purification devices and the method for a kind of fungi, particularly a kind of purifying is by the apparatus and method of the fungi of bacterial contamination.
Background technology:
At occurring in nature, mikrobe often is to mix to give birth to, and comprises the mixed life of fungi and bacterium, but in scientific research and production, needs the pure growth of single mikrobe.For by the radicula byssoidea of bacterial contamination, the method for existing removal bacterium has two types, and the first kind can add microbiotic etc. and chemically suppress bacterium in substratum.Second type, can use certain device to remove the bacterium on the fungi, method has 3, and 1, earlier fungi is connected on the agar plate, add the last layer agar medium then, mycelium can penetrate this layer agar medium in surface growth.Can separate bacterium and fungi like this.2, fungi is seeded on the agar-agar flat board, the deckglass that adds a slice sterilization then in the above compresses a little downwards, and making between deckglass and the substratum does not have the space, and the mycelium of fungi can pass agar medium and growth around deckglass.3, utilize glass ring to make fungi and bacterium separately; The glass ring that promptly uses with general spore germination experiment; Make somebody a mere figurehead central authorities with three thin short glass sticks at petridish; The mycelia of fungi just passes substratum and comes outside growing into below the glass ring, bacterium then is limited in the ring (Fang Zhongda: " planting the disease organon ").
But aforesaid method for severe contamination and with the removal of the compact bacterium of fungi, effect is bad.
Summary of the invention:
The present invention for solve that prior art exists to severe contamination and with the problem of the poor removal effect of the compact bacterium of fungi; The invention provides a kind of purifying by the apparatus and method of the filamentous fungus of bacterial contamination, purifying that can be effective, easy is by the filamentous fungus of bacterium severe contamination.
Technical scheme of the present invention is: a kind of purifying is by the device of the filamentous fungus of bacterial contamination; This device is formed by sealing film, purifying cup, purifying fungi collecting layer and biologically pure post; Seal film and cover the suitable for reading of purifying cup; Suspension is provided with the biologically pure post in the purifying cup, and the bottom of purifying cup is provided with purifying fungi collecting layer; The side ancient piece of jade, round, flat and with a hole in its centre of described biologically pure post and purifying cup, purifying fungi collecting layer, biology seal the gap of leaving 0.5～2cm between the film.The purifying cup can adopt containers such as beaker, glass cylinder or high-temperature resistance plastice cup.
The described film that seals is made up of the high-temperature resistance plastice film that is provided with the inoculation mouth in the middle of dustproof film and one deck, and inoculation mouthful diameter is less than purifying cup diameter suitable for reading, such as the aperture that is diameter 1～3cm.Described dustproof film can seal film or 6～8 layers gauze or 6～8 layers non-woven fabrics for tissue culture.
Described biologically pure post is made up of base material and fungus culture medium, and base material is a porous material, and base material and fungus culture medium are according to the mixed of 1: 1～2 (W/W).Described fungus culture medium is the mixture of plant tissue leach liquor, water, sugar.Sterilization as early as possible after processing, 112～115 ℃, 30min is for use in sterilization, or refrigeration (1～5 ℃) in short-term, and is subsequent use.Described base material is a kind of in dry branch, perlite, high temperature resistant sponge or the wood chip.If as base material, then dry branch is immersed fungus culture medium and gets final product after 10～24 hours with dry branch, the vertically vertical purifying cup bottom surface of branch.If when adopting perlite, high temperature resistant sponge or wood chip to prepare the biologically pure post, then with the ratio of 1: 1～2 (W/W) of fungus culture medium base material and fungus culture medium are mixed in base material.
Described purifying fungi collecting layer is that plant tissue leach liquor, water, agar and sugar are formed.The plant tissue leach liquor can be that plant lixiviate or squeeze the juice obtains.The plant tissue leach liquor is added entry, agar and sugar to be processed.Vegetable material can be selected piece root, stem tuber, fruit, cauline leaf of plant etc. for use.Used sugar is glucose or sucrose.Sterilization as early as possible after process purifying fungi collecting layer, 112～115 ℃, 30min is for use in sterilization, or refrigeration (1～5 ℃) in short-term, and is subsequent use.
After the assembling of described purification devices is accomplished, must be immediately with 112～115 ℃, sterilization 30min, purification devices is placed vertically in Autoclave during sterilization, the sterilization cooling that finishes is subsequent use.
Use described purifying by the method for the device purifying filamentous fungus of the filamentous fungus of bacterial contamination, step is:
The first step with the aseptic technique mode, is opened dustproof film, and contaminated filamentous fungus is gone into the top of biologically pure post from an inoculation mouthful inoculation, covers dustproof film again, and the purification devices of having inoculated fungi is cultivated down at 20～25 ℃;
Second step; Treat fungi when the biologically pure column length goes out and grow in purifying fungi collecting layer, open with the aseptic technique mode and seal film, the biologically pure post is taken away or kept; Fungi on the purifying fungi collecting layer is inoculated into inclined-plane or flat board, accomplish the purifying of filamentous fungus.
Principle: the present invention utilizes filamentous fungus and bacterium to the selective use of substratum nutriment with in the difference of storeroom growth characteristics; With filamentous fungus and with bacterium together be inoculated on the special biologically pure post; Growth through for some time; Bacterium is stayed the biologically pure post, and fungi grows the biologically pure post and grow in purifying fungi collecting layer, thereby reaches easily and effectively purifying by the purpose of the fungi of bacterial contamination.
Beneficial effect:
(1) can be reached perfectly sterile by the fungi of bacterial contamination after with method and apparatus purifying of the present invention, be particularly suitable for by the purifying of the fungi of bacterium severe contamination.
(2) the purification devices making method is simple, and manufacturing materials is cheap and easy to get.
(3) purifying to fungi sample in enormous quantities more can embody efficiently.
Description of drawings:
Fig. 1 is a purification devices synoptic diagram of the present invention,
Wherein 1 seal film, 2 inoculation mouths, 3 inoculation mouthful exterior seal rings, 4 inoculation mouthful inner seal rings, 5 biologically pure post messenger wires, 6 biologically pure posts, 7 purifying cups, 8 purifying fungi collecting layers.
Embodiment:
Below combine accompanying drawing that the present invention is further described through concrete examples of implementation.
1 makes purification devices
(1) produces purifying fungi collecting layer: select for use yam to produce the plant tissue leach liquor.With peeling potatoes, be cut into the little fourth of about 1.0cm * 1.0cm, in yam and tap water ratio with 1: 5～10 (W/W); Boiling, boiling 30min is with the fried liquid of filtered through gauze; Add water to the original water yield; Again by the fried liquid of yam: sucrose: agar dissolves with the mixed of 100: 2: 1.5 (W/W), the solid fungi culture medium, purifying fungi collecting layer just.Sterilization as early as possible after preparation is accomplished, 115 ℃ of sterilization 30min can be for use, or refrigeration (1～5 ℃) in short-term, and are subsequent use.。
(2) produce fungus culture medium: compare with the solid fungi culture medium, just need not add agar, just in the fried liquid of yam: sucrose promptly obtains fungus culture medium with the ratio preparation of 100: 2 (W/W), and the sterilization back is subsequent use.
(3) make the biologically pure post: cut-off is the tree section of the long 2～4cm of 1.5～3cm directly, if live tree section or humidity, then oven dry in advance is submerged into 10～24h in the fungus culture medium, for the control microorganism growth, can refrigerate immersion liquid.
(4) glass cylinder of cut-off footpath 5cm, high 7cm is as purifying cup 7; Put into the purifying fungi collecting layer 8 of thick about 0.5～2cm; Be around on the biologically pure post 6 as biologically pure post messenger wire 5 with commercially available 20～No. 22 zinc-plated thin wires; Biologically pure post 6 is hung in the purifying cup 7; Vertically erect and hang in the purifying cup 7, biologically pure post 6 is apart from the side ancient piece of jade, round, flat and with a hole in its centre of purifying cup 7, the surface and the purifying rim of a cup 0.5～2cm of purifying fungi collecting layer 8, with the film clad of resistant to elevated temperatures plastics film such as commercially available freshness protection package at the purifying rim of a cup; Mouth is tightened as inoculation mouthful inner seal ring 4 with rubber ring; Above purification column, cut or burn aperture and form inoculation mouthfuls 2, cover again with 6～8 layers gauze or 6～8 layers non-woven fabrics or tissue culture and seal film as dustproof film, with rubber ring as inoculate mouth exterior seal ring 3 will prevent dust film tighten purifying cup 7 mouthful on.The purification devices that assembles is sterilized as early as possible, 112～115 ℃ the sterilization 30min, purification devices is placed vertically in Autoclave during sterilization, the sterilization finish the cooling subsequent use.
2 purification process
Get the above-mentioned purification devices of processing, open dustproof film, fungal cultures to be purified is inoculated on the biologically pure post from the inoculation mouth, cover dustproof film once more, the purifying cup is put in 20～25 ℃ cultivates down.Treat that fungi stretches out from biologically pure post bottom, and after growing on the purifying fungi collecting layer, open and seal film 1, the biologically pure post is taken away or kept, the fungi on the purifying fungi collecting layer is inoculated into inclined-plane or flat board, accomplish the purifying of filamentous fungus.Adopt the fungi of the acquisition purifying that aforesaid method can be easily and effectively.
Similar with embodiment 1, what they were different is the making method of biologically pure post 6.Select for use the perlite boiling tap water to dry after 10 minutes or dry; The fungus culture medium that adds 1: 1 (W/W) is filled in the thick high-temperature resistance plastice pipe of 1.5～2.5cm, and the pipe end one end encases with the resistant to elevated temperatures net of moulding; Prevent that perlite from flowing out, and adopts this method can obtain the fungi of purifying.
Similar with embodiment 1, what they were different is the making method of solid fungi culture medium and fungus culture medium.Get soybean sprout and grind, extrude bean sprouts juice, add bean sprouts juice, sugar and tap water in 1: 5: 100 ratio and process fungus culture medium, select common edible sucrose for use with cloth.In fungus culture medium, add 2% (W/W) agar and process the solid fungi culture medium.Adopt this method can obtain the fungi of purifying.
Similar with embodiment 1, what they were different is the making method of solid fungi culture medium and fungus culture medium.Get apple or pears are squeezed the juice, after the filtered through gauze, add Sucus Mali pumilae or pear juice and tap water in 1: 10 ratio and process fungus culture medium, not sugaring.In fungus culture medium, add 2% (W/W) agar and process the solid fungi culture medium.Adopt this method can obtain the fungi of purifying.
Similar with embodiment 1, what they were different is that the film 8 that seals of fungi purification devices is selected 8 layers of gauze for use.Adopt this method can obtain the fungi of purifying.
Claims (4)
1. a purifying is by the device of the filamentous fungus of bacterial contamination; It is characterized in that; This device is formed by sealing film (1), purifying cup (7), purifying fungi collecting layer (8) and biologically pure post (6); Seal film (1) and cover the suitable for reading of purifying cup (7), suspension is provided with biologically pure post (6) in the purifying cup (7), and the bottom of purifying cup (7) is provided with purifying fungi collecting layer (8);
The described film (1) that seals is made up of the high-temperature resistance plastice film that is provided with inoculation mouthful (2) in the middle of dustproof film and one deck, and inoculation mouthful diameter (2) is less than purifying cup (7) diameter suitable for reading;
Described purifying fungi collecting layer (8) is that plant tissue leach liquor, water, agar and sugar are formed;
Described biologically pure post (6) is made up of base material and fungus culture medium; Described base material is any one in dry branch, perlite, high temperature resistant sponge or the wood chip: with dry branch during as base material; Dry branch is immersed fungus culture medium got final product in 10～24 hours, the vertically vertical purifying cup bottom surface of branch; , with the ratio of 1: 1～2 (W/W) of fungus culture medium base material and fungus culture medium are mixed during with perlite, high temperature resistant sponge or wood chip in base material as base material.
2. a kind of purifying as claimed in claim 1 is characterized in that by the device of the filamentous fungus of bacterial contamination described dustproof film is that tissue culture is sealed film or 6～8 layers gauze or 6～8 layers non-woven fabrics.
3. a kind of purifying as claimed in claim 1 is characterized in that by the device of the filamentous fungus of bacterial contamination the sidewall of described biologically pure post (6) and purifying cup (7), purifying fungi collecting layer (8), biology seal the gap of leaving 0.5～2cm between the film (1).
4. use purifying as claimed in claim 1 by the method for the device purifying filamentous fungus of the filamentous fungus of bacterial contamination, it is characterized in that step is:
The first step with the aseptic technique mode, is opened dustproof film, and contaminated filamentous fungus is gone into the top of biologically pure post (6) from inoculation mouthful (a 2) inoculation, covers dustproof film again, and the purification devices of having inoculated fungi is cultivated down at 20～25 ℃;
Second step; Treat that fungi grows and in purifying fungi collecting layer (8) when growing from biologically pure post (6); Open with the aseptic technique mode and to seal film (1); Biologically pure post (6) is taken away or kept, the fungi on the purifying fungi collecting layer (8) is inoculated into inclined-plane or flat board, accomplish the purifying of filamentous fungus.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 200910235086 CN101701179B (en) | 2009-11-13 | 2009-11-13 | Device and method for purifying mycelial fungi polluted by bacteria |
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| Application Number | Priority Date | Filing Date | Title |
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| CN 200910235086 CN101701179B (en) | 2009-11-13 | 2009-11-13 | Device and method for purifying mycelial fungi polluted by bacteria |
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| Publication Number | Publication Date |
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| CN101701179A CN101701179A (en) | 2010-05-05 |
| CN101701179B true CN101701179B (en) | 2012-01-04 |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN110699260A (en) * | 2019-09-16 | 2020-01-17 | 浙江省农业科学院 | Method for removing bacterial pollution in fungi |
| CN113428974B (en) * | 2021-06-18 | 2023-01-10 | 黑龙江佰瑞德环境生物科技有限公司 | Membrane bioreactor and method for slowing down membrane pollution of membrane bioreactor |
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Non-Patent Citations (2)
| Title |
|---|
| 徐士菊等.一种简便的微生物载片培养法.《生物学教学》.1980,(第01期),35、37. * |
| 陈碧娥等.从文蛤分离丝状真菌.《华侨大学学报(自然科学版)》.2006,第27卷(第01期),86-88. * |
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Address after: 212028 Zhenjiang, Dantu Metro Industrial Park Rui East Road, No. 9 Patentee after: Jiangsu University of Science and Technology Address before: 212003 Zhenjiang,, Jiangsu, Jiangsu University of Science and Technology, No. Patentee before: Jiangsu University of Science and Technology |
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