CN104611239A - Culture collection method for fungi mycelium - Google Patents
Culture collection method for fungi mycelium Download PDFInfo
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- CN104611239A CN104611239A CN201510073941.2A CN201510073941A CN104611239A CN 104611239 A CN104611239 A CN 104611239A CN 201510073941 A CN201510073941 A CN 201510073941A CN 104611239 A CN104611239 A CN 104611239A
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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 culture collection method for fungi mycelium. The culture collection method comprises the following steps: tiling glass paper on a culture medium of fungi growth, inoculating a fungal strain block on the glass paper in a sterile manner, and collecting fungi mycelium when the mycelia are grown in the culture medium. An applicant accidently finds that during the culture of fungi mycelium, the glass paper is taken as an isolating film, so that the fungal strain block is absorbed to the culture medium to nutritively and normally grow due to the gas permeability of the glass paper, the transparency of the glass paper is convenient for observing the appearances of the mycelia, and the mycelia and the culture medium are physically isolated to conveniently collect the mycelia without impurities. According to lots of experiments, the mycelia obtained by the culture collection method are suitable for extraction of total DNA, RNA or proteins and the like. The culture collection method is simple to operate, short in period and low in contamination rate; the obtained mycelia are pure and impurity-free; the glass paper is strong in easy decomposability, low in cost, free of environmental pollution and worthy of being popularized and used on a large scale.
Description
Technical field
The invention belongs to microorganism field, relate to a kind of cultivation collection method of radicula byssoidea.
Background technology
Researchist, in the extraction carrying out the STb gene of radicula byssoidea, RNA and protein or the research such as fungi fermentation formula, parameter exploration, needs to carry out cultivation propagation to mycelium.At present, the normal liquid fermentation and culture that adopts of cultivation propagation is carried out to mycelium and then mycelium is separated from fermented liquid, or direct test tube slant or the solid medium such as dull and stereotyped carry out the surperficial mycelial method of scraping after bacteria.
At present, mycelium is separated by mycelium many employings supercentrifugal process of liquid culture and filtration method from fermented liquid.But all there is certain problem in these two kinds of separation methods.For supercentrifugal process, on the one hand, due to the mycelium comparatively thickness of some fungi strain liquid culture, become suspension macrobead state, therefore, even if adopt very high centrifugal speed, also be difficult to mycelium to be deposited to completely compactly bottom centrifuge tube, inferior separating effect; And on the other hand, the moisture of mycelium pellet granule interior is also difficult to by centrifugal segregation, and there is the problems such as whizzer cost is high, complex operation is consuming time.For filtration method, comprise at present and adopt filter paper to carry out the method for vacuum filtration and common filtered through gauze method, for filter paper vacuum filtration method, on the one hand, same existence is due to the high and thickness of the concentration of some fungi strain fermented liquid, be difficult to the problem obtaining drier mycelia, on the other hand, for such fermented liquid, filter paper micropore is very easily fermented liquid and blocks, therefore, follow-uply be difficult to filtration and go down, constantly must change filter paper, and due to filter paper wet after easily scratched, be therefore adsorbed on that mycelium on filter paper is also more difficult completely to be taken off; For filtered through gauze method, because gauze is easily stained with mycelia, therefore, easily cause the mycelia crossed contamination of different sample room when sample size is many, not reproducible utilization, thus cause unnecessary waste, and when adopting filtered through gauze, the gauze number of plies at least mycelia easily spills, and the number of plies easily remains mycelia between every layer of gauze and in gauze hole at most, easily causes the waste of sample and the error of data.
And for the mycelial method in scraping surface after the solid culture such as test tube slant or flat board mycelium, hyphae length is too little is on the one hand difficult to satisfied experiment demand, on the other hand, solid medium is brought into unavoidably in scraping mycelium process, not only troublesome poeration is loaded down with trivial details, and affects carrying out and the accuracy of experimental result of follow-up molecular biosciences experiment.
Summary of the invention
For above deficiency, the radicula byssoidea that the invention provides a kind of simple and fast is cultivated and collection method, not only makes mycelial collection convenient and simple, and the pure nothing of collected mycelium is mixed, and is suitable for molecular biology research.
The present invention is achieved the above object by following scheme:
A cultivation collection method for radicula byssoidea, is laid in glassine paper on the substratum of fungal growth, by fungi strain block aseptic inoculation on glassine paper, covers with after substratum collect radicula byssoidea until mycelium.
Preferably, cover with after substratum until mycelium, glassine paper is separated with substratum, collect the mycelium on glassine paper.
Preferred further, after being separated with substratum by glassine paper, the mycelium on scraping glassine paper is collected and is obtained mycelium.
Preferably, described fungi strain block inoculation size is between (0.5 ± 0.2) cm* (0.5 ± 0.2) cm.
Preferably, described substratum is culture medium flat plate.
Preferably, described mycelium is used for the extraction of STb gene, RNA or protein.
Glassine paper (Cellophane), also known as cellulose film, is the high and glossiness RCF regenerated cellulose film of a kind of transparency, and to the fresh-keeping of commodity with to preserve activity very favourable, therefore glassine paper is often used as commodity packaging at present.
Applicant surprisingly finds, when hypha,hyphae is cultivated, when particularly utilizing fungi strain block to inoculate, glassine paper is used as barrier film, its permeability makes fungi kind block can absorb the nutrition of substratum and normal growth through it, and the transparency of glassine paper is not only convenient to observe mycelium outward appearance; And mycelium and substratum physical isolation can be opened, be convenient to mycelial collection and do not bring impurity into.Through experimental results demonstrate, this cultivation collection method collects the mycelium obtained, and is suitable for follow-up for STb gene, RNA or the biological study of proteins extraction equimolecular.This cultivation collection method is simple to operate, and the cycle is short, and pollution rate is low, the pure inclusion-free of mycelium of acquisition, and glassine paper has very strong easy decomposability, and cost is low and can not cause environmental pollution, is worth large-scale promotion to utilize.
Accompanying drawing explanation
Fig. 1 is the electrophorogram of the ITS-PCR product of embodiment 4.
Embodiment
Below in conjunction with specific embodiment, the present invention is further described.
Embodiment 1:
A kind of cultivation collection method of radicula byssoidea, glassine paper is laid on the culture medium flat plate of fungal growth, by fungi strain block aseptic inoculation on the glassine paper on culture medium flat plate, covers with after culture medium flat plate until mycelium, glassine paper is taken out, collects the mycelium on glassine paper.
Embodiment 2:
A kind of cultivation collection method of radicula byssoidea, by glassine paper sterilizing, prepare the culture medium flat plate being applicable to hypha,hyphae bulk-growth, after culture medium solidifying, glassine paper after sterilizing is laid on culture medium flat plate, aseptic access fungi strain block, build culture medium flat plate lid, postvaccinal flat board is placed in the environment of applicable fungal growth, period observes mycelial growth situation, if pollution condition, abandon, until after radicula byssoidea covers with culture medium flat plate, glassine paper is taken out from culture medium flat plate, be placed in new vessel, mycelium on scraping glassine paper, obtain the radicula byssoidea collected.
Embodiment 3:
A kind of cultivation collection method of radicula byssoidea:
1. colourless transparent glass paper is cut into the circle more smaller than culture dish diameter, wrapping sterilizing is for subsequent use;
2. carried out being down flat plate by the preparation radicula byssoidea medium liquid that also sterilizing is good in super clean bench, tile last layer glassine paper after substratum plane is slightly solidified, and obtains the culture medium flat plate with glassine paper;
3. on the glassine paper of culture medium flat plate, aseptic access size is the fungi kind block of (0.5 ± 0.2) cm* (0.5 ± 0.2) cm, builds flat plate cover, and uses ParafilmTM.
4. postvaccinal flat board is placed in the environment being suitable for fungal growth to grow, until mycelium covers with flat board;
5. provoke glassine paper gently with tweezers to be transferred in new empty culture dish ware, mycelium and substratum are easily separated;
6., by the empty culture dish that the mycelium scraping on glassine paper is extremely new, 35-60 DEG C of oven dry saves backup.
Often open 10 yuan of calculating with the glassine paper of 100*120cm, often open glassine paper and be cut into 120-140 and open the diaphragm being applicable to culture medium flat plate, therefore the method cost is low.
Embodiment 4:
1. the glassine paper of 100*120cm is cut into the diameter circle more smaller than culture dish, one deck glassine paper one deck filter paper is placed in culture dish, and it is for subsequent use to wrap sterilizing;
2. carried out being down flat plate by the preparation radicula byssoidea substratum that also sterilizing is good in super clean bench, tile the sterilized glassine paper of last layer after substratum plane is slightly solidified, and obtains the culture medium flat plate with glassine paper;
3. on the glassine paper of culture medium flat plate, aseptic access size is the fungi kind block of (0.5 ± 0.2) cm* (0.5 ± 0.2) cm, builds flat plate cover, and with ParafilmTM;
4. postvaccinal flat board is placed in the environment bacteria being suitable for this hypha,hyphae bulk-growth, until mycelium covers with flat board;
5. provoke glassine paper gently with the tweezers of sterilizing to be transferred in new empty culture dish, mycelium and substratum can be made easily to separate;
6., by the empty culture dish that the mycelium scraping on glassine paper is extremely new, 35-60 DEG C of oven dry saves backup, and namely completes cultivation and the collection of radicula byssoidea.
Above experimental procedure is used to carry out Mycelium culture collection to 10 fungies, and total DNA extraction is carried out to each mycelium, ITS-PCR is carried out to each mycelium STb gene, the list of 10 fungies is as table 1, the primer of ITS-PCR is:, ITS1:TCC GTA GGT GAA CCT GCG G and ITS4:TCC TCC GCT TAT TGA TAT GC, the product of ITS-PCR is carried out electrophoresis, and electrophoretogram as shown in Figure 1.
From Fig. 1 mycelial ITS-PCR product gel electrophorogram, it is effective that application the method cultivates the mycelium collected, and is suitable for the meticulous experimental studies such as molecular biology.Adopt this cultivation collection method to cultivate and collect mycelium, simple and quick, pollution rate is low, and mycelia yield is high, and gained mycelium is pure.
Table 1:
The above; be only preferably specific embodiment of the present invention; but protection scope of the present invention is not limited thereto; anyly be familiar with those skilled in the art in the technical scope that the present invention discloses; be equal to according to technical scheme of the present invention and design thereof and replace or change, all should be encompassed in protection scope of the present invention.
Claims (6)
1. a cultivation collection method for radicula byssoidea, is characterized in that, is laid in by glassine paper on the substratum of fungal growth, by fungi strain block aseptic inoculation on glassine paper, covers with after substratum collect radicula byssoidea until mycelium.
2. the cultivation collection method of a kind of radicula byssoidea as claimed in claim 1, is characterized in that, covers with after substratum, be separated by glassine paper with substratum until mycelium, collects the mycelium on glassine paper.
3. the cultivation collection method of a kind of radicula byssoidea as claimed in claim 1, is characterized in that, after being separated with substratum by glassine paper, the mycelium on scraping glassine paper is collected and obtained mycelium.
4. the cultivation collection method of a kind of radicula byssoidea as claimed in claim 1, is characterized in that, described fungi strain block inoculation size is between for (0.5 ± 0.2) cm* (0.5 ± 0.2) cm.
5. the cultivation collection method of a kind of radicula byssoidea as claimed in claim 1, is characterized in that, described substratum is culture medium flat plate.
6. the cultivation collection method of a kind of radicula byssoidea as claimed in claim 1, is characterized in that, described mycelium is used for the extraction of STb gene, RNA or protein.
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Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105766205A (en) * | 2016-04-01 | 2016-07-20 | 湖北未来家园高科技农业有限公司 | Harvesting method of Cordyceps taishanensis |
| CN106612982A (en) * | 2016-12-21 | 2017-05-10 | 广东省微生物研究所 | Efficient pleurotus tuber-regium multi-spore isolation method and multi-spore isolation device used in same |
| CN108715813A (en) * | 2018-05-25 | 2018-10-30 | 福建农林大学 | A method of collecting pure mycelia |
| CN109797160A (en) * | 2018-09-26 | 2019-05-24 | 天津科技大学 | The construction method of heterogenous expression exoglucanase Cel6A, Cel7A in a kind of Pichia pastoris |
| CN109971784A (en) * | 2018-09-26 | 2019-07-05 | 天津科技大学 | Heterogenous expression endoglucanase EG II in a kind of Pichia pastoris, the construction method of EG IV, EG V |
| CN110616152A (en) * | 2019-09-20 | 2019-12-27 | 广东省微生物研究所（广东省微生物分析检测中心） | Preparation method of improved fungus protoplast |
| CN112375689A (en) * | 2020-10-28 | 2021-02-19 | 中国科学院天津工业生物技术研究所 | Hypha culture method and method for preparing and regenerating protoplast by using hypha culture method |
| CN113549556A (en) * | 2021-06-10 | 2021-10-26 | 甘肃农业大学 | Method for collecting mycelium of flat plate liquid culture fungus |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101434997A (en) * | 2008-12-23 | 2009-05-20 | 华中农业大学 | Early stage rapid molecular detection method for rape sclerotinia rot |
| CN101434630A (en) * | 2008-11-17 | 2009-05-20 | 山东省农业科学院土壤肥料研究所 | Method for extracting mushroom genome |
| CN103571760A (en) * | 2013-11-13 | 2014-02-12 | 华南农业大学 | Separation and purification method of beauveria bassiana |
-
2015
- 2015-02-11 CN CN201510073941.2A patent/CN104611239A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101434630A (en) * | 2008-11-17 | 2009-05-20 | 山东省农业科学院土壤肥料研究所 | Method for extracting mushroom genome |
| CN101434997A (en) * | 2008-12-23 | 2009-05-20 | 华中农业大学 | Early stage rapid molecular detection method for rape sclerotinia rot |
| CN103571760A (en) * | 2013-11-13 | 2014-02-12 | 华南农业大学 | Separation and purification method of beauveria bassiana |
Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105766205A (en) * | 2016-04-01 | 2016-07-20 | 湖北未来家园高科技农业有限公司 | Harvesting method of Cordyceps taishanensis |
| CN106612982A (en) * | 2016-12-21 | 2017-05-10 | 广东省微生物研究所 | Efficient pleurotus tuber-regium multi-spore isolation method and multi-spore isolation device used in same |
| CN108715813A (en) * | 2018-05-25 | 2018-10-30 | 福建农林大学 | A method of collecting pure mycelia |
| CN109797160A (en) * | 2018-09-26 | 2019-05-24 | 天津科技大学 | The construction method of heterogenous expression exoglucanase Cel6A, Cel7A in a kind of Pichia pastoris |
| CN109971784A (en) * | 2018-09-26 | 2019-07-05 | 天津科技大学 | Heterogenous expression endoglucanase EG II in a kind of Pichia pastoris, the construction method of EG IV, EG V |
| CN110616152A (en) * | 2019-09-20 | 2019-12-27 | 广东省微生物研究所（广东省微生物分析检测中心） | Preparation method of improved fungus protoplast |
| CN112375689A (en) * | 2020-10-28 | 2021-02-19 | 中国科学院天津工业生物技术研究所 | Hypha culture method and method for preparing and regenerating protoplast by using hypha culture method |
| CN113549556A (en) * | 2021-06-10 | 2021-10-26 | 甘肃农业大学 | Method for collecting mycelium of flat plate liquid culture fungus |
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