CN102146343B - Storage method capable of keeping activity of nematode-trapping fungi for long time - Google Patents
Storage method capable of keeping activity of nematode-trapping fungi for long time Download PDFInfo
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- CN102146343B CN102146343B CN2010106170008A CN201010617000A CN102146343B CN 102146343 B CN102146343 B CN 102146343B CN 2010106170008 A CN2010106170008 A CN 2010106170008A CN 201010617000 A CN201010617000 A CN 201010617000A CN 102146343 B CN102146343 B CN 102146343B
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Abstract
The invention discloses a storage method capable of keeping the activity of nematode-trapping fungi for a long time, which mainly comprises: performing high-pressure sterilization of water-saturated natural soil by using a centrifugal tube; and transferring a strain to be stored onto the surface of soil in the centrifugal tube for cultivation and storage. In the invention, the water-saturated soil, of which the water condition is close to that of the strain in a culture medium and in which the soil is closer to the natural living environment of the nematode-trapping fungi, is used to store the strain, so the storage time is long. Therefore, the problem of lowering the food hunting ability of the fungi due to multiple times of tube transfer of the conventional storage method is solved, the fungus-containing soil can be repeatedly used in a storage period, the drawback that the mites eat fungi and pollute soil, the drawback that the storage time is short and other drawbacks of the conventional method are overcome, and a convenience is brought to the study of the strain.
Description
Technical field
The present invention relates to the active store method of a kind of long-term maintenance nematode-trapping fungi.Be applicable to the Plant nematode biocontrol microorganisms, researchs such as animal parasitic nematode biocontrol microorganisms and microbiology.
Background technology
Occurring in nature exist a part can prey on, parasitic or grow the fungi on nematode surely, this type fungi is a Nematophagous fungi.Wherein pass through the trapping organs such as the viscosity mycelia of vegetative hyphae specialization, the viscosity branch, the viscosity net, the viscosity ball, shrunk ring and non-shrunk ring are caught nematode, and this type Nematophagous fungi is a nematode-trapping fungi.Nematode-trapping fungi is the one type of biocontrol microorganisms that the control of plant nematode is had potential ability.But the researchist when isolated bacterial strain is further studied, relates to the preservation problem of bacterial classification in the process of research.Therefore, a good store method has great importance to its research.
Preservation method commonly used is the test tube slant substratum.Its method is with waiting that preserving bacterial classification inserts inclined-plane in PDA or the Corn Meal Agar test tube substratum, cultivated 2-3 days down at 25 ℃, placing then under 4 ℃ of conditions and preserve.The deficiency that this store method exists is: the shelf time is short, is merely half a year, and is easy to receive the mite class to pollute, and it is big to take up room, and is prone to fragmentation, not portable, and the tube frequency is high, and cost is high relatively, is prone to reduce the prey ability of fungi, influences progress.
Summary of the invention
The objective of the invention is to overcome the deficiency of conventional store method; Provide a kind of long-term maintenance nematode-trapping fungi active store method; The natural soils that utilize saturation moisture content prolonged preservation nematode-trapping fungi in Eppendorf tube especially in centrifuge tube, nematode-trapping keeps its activity near under the natural condition; Prolong the shelf time, thereby be used for further research.
For achieving the above object, the technical scheme that the present invention takes is:
The first step: the preparation of substratum
The natural soils of the saturation moisture content of 1/3rd pipe volume of in centrifuge tube, packing into wrap up centrifuge tube with aluminium foil, under 121 ℃ of conditions to centrifuge tube and inner soil autoclaving 1 hour at once.
Second step: wait to deposit culture of strains and preservation
Under aseptic condition, the bacterial classification that needs are preserved changes the soil surface in the centrifuge tube over to, covers tight lid, cultivates 2 days down at 25 ℃, cultured bacterial classification is preserved under 4 ℃ of conditions together with centrifuge tube to get final product then.
In order to make the preservation bacterial classification be convenient to preserve and carry, described centrifuge tube is the Eppendorf tube (English name eppendorf pipe is called for short the EP pipe) of 1.5ml.
Advantage of the present invention is:
1, through the soil that utilizes saturation moisture content bacterial classification is preserved, its condition is near the moisture condition of bacterial classification at substratum, and soil wherein is more near the natural living environment of nematode-trapping fungi, so the shelf time is long.
2, avoided existing store method to reduce the prey ability of fungi because of tube repeatedly; The soil that during preservation carries disease germs in the pipe can repeatedly use; Overcome conventional store method and be prone to got the food pollution by the mite class, deficiencies such as shelf time weak point are provided convenience for studying this bacterial classification.
3, utilize EP to guarantee to deposit portable, light, volume is little, is difficult for brokenly, is difficult for polluting, and it is wide to preserve the matrix source, is prone to obtain, and cost is low, long preservative period.
In order to verify the vigor of store method bacterial classification of the present invention; Under equal conditions utilize store method of the present invention and existing store method to compare test; The result finds to utilize store method of the present invention that active preservation of nematode-trapping fungi kept 1 year at least; And utilize existing store method that active preservation of nematode-trapping fungi only kept half a year, this explanation the inventive method is preserved and is much better than existing store method.
The practical implementation method
Specify the step of store method of the present invention below in conjunction with specific embodiment.
The first step: the soil of preparation saturation moisture content
One deck qualitative filter paper along funnel internal surface shop lets filter paper suction water, and soil is put into funnel inside (not ultra-filtration trimming edge), adds water gradually, till there is a water at filter paper knuckle place and does not drip.
Second step: utilize saturation moisture content soil processing substratum
The soil for preparing is changed in the 1.5ml eppendorf pipe (being called for short the EP pipe) with spoon; The amount of soil accounts for 1/3rd of EP pipe volume, and EP is uncovered with the aluminium foil parcel, carefully puts into high-pressure sterilizing pot;, under 121 ℃ of temperature, 20 pounds of pressure conditions, sterilized 1 hour.
The 3rd step: wait to deposit culture of strains and preservation
Under aseptic condition, the bacterial classification that needs are preserved changes soil surface in the EP pipe over to, covers tight lid, cultivates 2 days down at 25 ℃, puts it into refrigerator then and under 4 ℃ of conditions, preserves and get final product.
Claims (2)
1. one kind long-term keeps the active store method of nematode-trapping fungi, it is characterized in that step is following:
The first step: the preparation of substratum
The natural soils of the saturation moisture content of 1/3rd pipe volume of in centrifuge tube, packing into, with aluminium foil parcel EP pipe, under 121 ℃ of conditions to centrifuge tube and inner soil autoclaving 1 hour;
Second step: wait to deposit culture of strains and preservation
Under aseptic condition, the bacterial classification that needs are preserved changes the soil surface in the centrifuge tube over to, covers tight lid, cultivates 2 days down at 25 ℃, cultured bacterial classification is preserved under 4 ℃ of conditions together with centrifuge tube to get final product then.
2. the active store method of long-term maintenance nematode-trapping fungi as claimed in claim 1 is characterized in that step is following:
The first step: the soil of preparation saturation moisture content
One deck qualitative filter paper along funnel internal surface shop lets filter paper suction water, and soil is put into funnel inside and do not wanted ultra-filtration trimming edge, adds water gradually, till there is a water at filter paper knuckle place and does not drip;
Second step: utilize saturation moisture content soil processing substratum
The soil for preparing is changed in the 1.5ml Eppendorf tube with spoon, and the amount of soil accounts for 1/3rd of centrifuge tube volume, carefully puts into high-pressure sterilizing pot with aluminium foil parcel with centrifuge tube is uncovered, sterilizes 1 hour down at 121 ℃ of temperature, 20 pounds of pressure conditions;
The 3rd step: wait to deposit culture of strains and preservation
Under aseptic condition, the bacterial classification that needs are preserved changes soil surface in the EP pipe over to, covers tight lid, cultivates 2 days down at 25 ℃, puts it into refrigerator then and under 4 ℃ of conditions, preserves and get final product.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2010106170008A CN102146343B (en) | 2010-12-31 | 2010-12-31 | Storage method capable of keeping activity of nematode-trapping fungi for long time |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2010106170008A CN102146343B (en) | 2010-12-31 | 2010-12-31 | Storage method capable of keeping activity of nematode-trapping fungi for long time |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN102146343A CN102146343A (en) | 2011-08-10 |
| CN102146343B true CN102146343B (en) | 2012-07-25 |
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| Application Number | Title | Priority Date | Filing Date |
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| CN2010106170008A Expired - Fee Related CN102146343B (en) | 2010-12-31 | 2010-12-31 | Storage method capable of keeping activity of nematode-trapping fungi for long time |
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Families Citing this family (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106962324A (en) * | 2017-04-28 | 2017-07-21 | 广东省生物资源应用研究所 | A kind of method of long-term storage entomopathogenic nematode |
| CN113481108B (en) * | 2021-07-29 | 2023-09-12 | 大理大学 | Nutritional matrix for stimulating growth of nematode-trapping fungi on trunk, and preparation method and application method thereof |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1295125A (en) * | 1999-11-03 | 2001-05-16 | 中国科学院沈阳应用生态研究所 | Filamentous fungus passing, culturing, coating and preserving method |
| CN101283955A (en) * | 2007-04-13 | 2008-10-15 | 味之素株式会社 | Aqueous preservative solution with high amino acid content |
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2010
- 2010-12-31 CN CN2010106170008A patent/CN102146343B/en not_active Expired - Fee Related
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1295125A (en) * | 1999-11-03 | 2001-05-16 | 中国科学院沈阳应用生态研究所 | Filamentous fungus passing, culturing, coating and preserving method |
| CN101283955A (en) * | 2007-04-13 | 2008-10-15 | 味之素株式会社 | Aqueous preservative solution with high amino acid content |
Non-Patent Citations (1)
| Title |
|---|
| 根井外喜男.土壤保存法.《微生物保存法》.1982,正文第5页第12-21行，第6页第9-13行. * |
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| CN102146343A (en) | 2011-08-10 |
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Granted publication date: 20120725 Termination date: 20121231 |