CN101220344B - Filter paper sheet diaphragmatic foramen suspension spore ejection method - Google Patents
Filter paper sheet diaphragmatic foramen suspension spore ejection method Download PDFInfo
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- CN101220344B CN101220344B CN 200810070482 CN200810070482A CN101220344B CN 101220344 B CN101220344 B CN 101220344B CN 200810070482 CN200810070482 CN 200810070482 CN 200810070482 A CN200810070482 A CN 200810070482A CN 101220344 B CN101220344 B CN 101220344B
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Abstract
The invention discloses a filter paper separated hole suspended spore ejection method, which firstly prepares a sterile potato culture medium slab with the thickness of 0.5cm and processes a sterile filter paper which is provided with a hole with the diameter of Phi 0.5 to 2 cm; then a culture dish cover is removed from a super clean workbench, and the sterile filter paper with the hole is arranged on a lower dish which is provided with the culture medium slab; a large-scale fungal sporophore block which is collected at the field is arranged on the filter paper hole of the sterile filter paper with the hole for still placement for 20 to 60min, then the filter paper and the sporophore are removed, the culture dish cover is covered, and a constant temperature culture is carried out after transferring the fungal sporophore block into a culture box at 22 - 26 DEG C; finally, the mycelia are selected after spore germination and transferred into the slope of a test tube for continual culture till the tube is full; the configuration identification is further carried out, so as to obtain a wild large-scale fungal pure culture. The invention has the advantages of reducing the opportunity of hybrid bacteria pollution, improving the success rate of the obtainment of large-scale fungal pure culture and having simplified operation.
Description
Technical field
The present invention relates to the fungi strain separation and purification, thereby especially relate to a kind of filter paper sheet diaphragmatic foramen suspension spore ejection method that can separate acquisition macro fungi pure growth efficiently and easily.
Technical background
Separate the mother that the macro fungi pure growth is directly connected to bacterial classification and plant acquisitions, female kind quality, and aspect such as breeding material.The spore method of launching is applicable to that the scope of separating macro fungi is wider, and can obtain the monospore pure growth.But the spore method of launching is easy to pollution microbes such as mould or bacterium etc., and the assorted relatively large fungus breeding of bacterium of this class is fast etc. former thereby often cause the failure of pure culture in addition.The conventional spore method of launching has three kinds: whole mushroom inserts kind of a method, the outstanding method of hook and attaching method.Whole mushroom inserts kind of method to be needed special device collection spore and does the dilution coating, operates very loaded down with trivial details.The outstanding method of hook is the cap hook to be hanged make spore fall into the triangular flask substratum, easily pollutes, and is difficult for switching.The attaching method is with lamella or cap is attached to the inclined-plane or culture dish covers, and treats after spore falls substratum to be moved in test tube or the culture dish and cultivates, complex operation, easily microbiological contamination.
Summary of the invention
The purpose of this invention is to provide a kind of chance, raising that can reduce living contaminants and obtain the success ratio of macro fungi pure growth, the filter paper sheet diaphragmatic foramen suspension spore ejection method that simplifies the operation.
The object of the present invention is achieved like this:
1, prepare potato culture (PDA), make the flat board of thick 0.5cm after the sterilization, be processed with the circular filter paper hole that diameter is Φ 0.5～2cm simultaneously on filter paper, be prepared into the punching filter paper, it is standby to sterilize;
2, in Bechtop, take off the culture dish lid, aseptic punching filter paper is placed on the following ware that has culture medium flat plate, macro fungi sporophore piece with field acquisition places on the filter paper hole of aseptic punching filter paper again, leave standstill 20～60min, remove filter paper and sporophore, cover the culture dish lid, move to constant temperature culture in 22～26 ℃ of incubators;
3, treating that the picking mycelium is transferred to after the spore germination continues to be cultured to full packages in the test tube slant, further makes identification of morphology, obtains the macro fungi pure growth.
The punching filter paper handles by the high pressure moist heat sterilization or ultraviolet disinfection is handled.
Filter paper sheet diaphragmatic foramen suspension spore ejection method of the present invention is to improve to form on outstanding method of hook and attaching method basis.Therefore the present invention has following advantage:
1, significantly reduced the chance of living contaminants.One, this method are placed directly in the sporophore piece on the alternating floor aseptic filter paper with holes and allow its spore launch to culture medium flat plate from hole.With outstanding method of conventional hook and attaching method all sporophore pieces all are exposed to the overhead ratio of culture medium flat plate, have significantly reduced assorted bacterium and may cause the area that pollutes; Its two, this method is after the sporophore piece launches 20～60min, spore is not sprouted and is just removed sporophore.Remove sporophore than having significantly reduced the time that assorted bacterium may pollute after launching 24h with the outstanding method of hook; Its three, it is to carry out on the Bechtop of aseptic technique that this method solid block spore launches, and promptly removes the sporophore piece of the assorted bacterium of band subsequently.This compares with the attaching method with the hook method of hanging, and causes assorted bacterium to shake the chance fallen substratum on because of culture dish device (triangular flask or culture dish) moves concussion with assorted mushroom solid block than the major general in the spore ejection process.
2, make the spore method of launching obtain simplifying.This method is to avoid influencing because of the damage that sterilize halfway pollution or sterilizing agent permeate spore the effect of its sprouting, sporophore piece to be separated is not sterilized directly use.The spore ejection process is finished on the Bechtop of aseptic technique, remove filter paper and sporophore piece subsequently, rather than the aseptic technique of sporophore piece is suspended from triangular flask or the culture dish as outstanding method of hook or attaching method, after the immigration incubator launches about 24h by it, on Bechtop, remove the sporophore piece again.
Description of drawings
Fig. 1 carried out spore and launches isolating design sketch (with the mushroom is material, launches the back and cultivates 10d) for the difference time of launching;
Fig. 2 is the design sketch (40mina thickness be 1cm, 40minb thickness be 0.5cm) of different culture medium flat plate thickness to launching influential effect.
Wherein: Fig. 1 launches spore germination situation under the time for difference, and the time of launching is long more, and the sprouting amount is big more, but increase in time, and that dyes assorted bacterium may also increase (launch 60min and mould, bacterial contamination occur); 40mina/b is respectively thick about 1cm among Fig. 2, the 0.5cm culture medium flat plate launches 40min result: catapulting distance is near more, and the spore degree of scatter is low more.
Embodiment
Be described in further detail the present invention below in conjunction with embodiment, but should understand the scope that scope of the present invention is not limited only to these embodiment.
Embodiment 1:
1, prepares potato culture (PDA), make the flat board of thick 0.5cm after the sterilization, on filter paper, be processed with the circular filter paper hole that diameter is Φ 0.5cm simultaneously, be prepared into the punching filter paper, handle standby by ultraviolet disinfection;
2, in Bechtop, take off the culture dish lid, aseptic punching filter paper is placed on the following ware that has culture medium flat plate, macro fungi oak money bacterium [Collybia dryophila (Bull.Ex Fr.) Que`l] sporophore piece with field acquisition places on the filter paper hole of aseptic punching filter paper again, after leaving standstill 60min, remove filter paper and sporophore, cover the culture dish lid, move to constant temperature culture in 22 ℃ of incubators;
3, treating that the picking mycelium is transferred to after the spore germination continues to be cultured to full packages in the test tube slant, further makes identification of morphology, obtains oak money bacterium pure growth.
Embodiment 2:
1, prepares potato culture (PDA), make the flat board of thick 0.5cm after the sterilization, on filter paper, be processed with the circular filter paper hole that diameter is Φ 2cm simultaneously, be prepared into the punching filter paper, handle standby by the high pressure moist heat sterilization;
2, in Bechtop, take off the culture dish lid, aseptic punching filter paper is placed on the following ware that has culture medium flat plate, macro fungi oyster cap fungus [Pleurotus ostreatus (Jacq.Ex Fr.) Que`l] sporophore piece with field acquisition places on the filter paper hole of aseptic punching filter paper again, after leaving standstill 20min, remove filter paper and sporophore, cover the culture dish lid, move to constant temperature culture in 26 ℃ of incubators;
3, treating that the picking mycelium is transferred to after the spore germination continues to be cultured to full packages in the test tube slant, further makes identification of morphology, obtains the oyster cap fungus pure growth.
Embodiment 3:
1, prepares potato culture (PDA), make the flat board of thick 0.5cm after the sterilization, on filter paper, be processed with the circular filter paper hole that diameter is Φ 1cm simultaneously, be prepared into the punching filter paper, handle standby by ultraviolet disinfection;
2, in Bechtop, take off the culture dish lid, aseptic punching filter paper is placed on the following ware that has culture medium flat plate, the sticking rust of macro fungi ear [Crepidotus mollis (Fr.) Staud] sporophore piece with field acquisition places on the filter paper hole of aseptic punching filter paper again, after leaving standstill 40min, remove filter paper and sporophore, cover the culture dish lid, move to constant temperature culture in 24 ℃ of incubators;
3, treating that the picking mycelium is transferred to after the spore germination continues to be cultured to full packages in the test tube slant, further makes identification of morphology, obtains sticking rust ear pure growth.
Claims (2)
1. filter paper sheet diaphragmatic foramen suspension spore ejection method is characterized in that:
(1), prepare the PDA substratum, make the flat board of thick 0.5cm after the sterilization, on filter paper, be processed with the circular filter paper hole that diameter is Φ 0.5～2cm simultaneously, be prepared into the punching filter paper, it is standby to sterilize;
(2), in Bechtop, take off the culture dish lid, aseptic punching filter paper is placed on the following ware that has culture medium flat plate, macro fungi sporophore piece with field acquisition places on the filter paper hole of aseptic punching filter paper again, leave standstill 20～60min, remove filter paper and sporophore, cover the culture dish lid, move to constant temperature culture in 22～26 ℃ of incubators;
(3), treat that the picking mycelium is transferred to after the spore germination and continue to be cultured to full packages in the test tube slant, further make identification of morphology, obtain the macro fungi pure growth.
2. filter paper sheet diaphragmatic foramen suspension spore ejection method as claimed in claim 1 is characterized in that: the punching filter paper handles by the high pressure moist heat sterilization or ultraviolet disinfection is handled.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 200810070482 CN101220344B (en) | 2008-01-15 | 2008-01-15 | Filter paper sheet diaphragmatic foramen suspension spore ejection method |
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| Application Number | Priority Date | Filing Date | Title |
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| CN 200810070482 CN101220344B (en) | 2008-01-15 | 2008-01-15 | Filter paper sheet diaphragmatic foramen suspension spore ejection method |
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| CN101220344A CN101220344A (en) | 2008-07-16 |
| CN101220344B true CN101220344B (en) | 2011-05-04 |
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Families Citing this family (5)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN104830697B (en) * | 2015-04-29 | 2017-12-05 | 潍坊科技学院 | A kind of preparation method of thread spore fungal cultures dry preserved specimen |
| CN104745522B (en) * | 2015-04-29 | 2018-05-25 | 潍坊科技学院 | A kind of method for inducing silk spore fungi spore |
| CN109197398B (en) * | 2018-11-09 | 2021-07-09 | 贵州大学 | Method for collecting clean basidiospores of agaricus fungus |
| CN110564628B (en) * | 2019-09-29 | 2020-12-29 | 遵义市林业科学研究所 | Total nutrient substrate microorganism culture method |
| CN111849788A (en) * | 2020-08-04 | 2020-10-30 | 青海省农林科学院 | Preservation method of barley stripe disease germs and co-culture filter paper sheet |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP1294850B1 (en) * | 2000-05-02 | 2004-07-21 | Young-Ryun Chung | A microbial pesticide active against plant fungal pathogens and process for preparation thereof |
| CN1786149A (en) * | 2004-12-08 | 2006-06-14 | 中国科学院沈阳应用生态研究所 | Field separation and activity preservation method of agaric spore |
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Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP1294850B1 (en) * | 2000-05-02 | 2004-07-21 | Young-Ryun Chung | A microbial pesticide active against plant fungal pathogens and process for preparation thereof |
| CN1786149A (en) * | 2004-12-08 | 2006-06-14 | 中国科学院沈阳应用生态研究所 | Field separation and activity preservation method of agaric spore |
Non-Patent Citations (1)
| Title |
|---|
| 胡建伟等.巴楚蘑菇孢子弹射与收集方法的研究.中国食用菌22 5.2002,22(5),15-16. * |
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