CN1295125A - Filamentous fungus passing, culturing, coating and preserving method - Google Patents
Filamentous fungus passing, culturing, coating and preserving method Download PDFInfo
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- CN1295125A CN1295125A CN 99113397 CN99113397A CN1295125A CN 1295125 A CN1295125 A CN 1295125A CN 99113397 CN99113397 CN 99113397 CN 99113397 A CN99113397 A CN 99113397A CN 1295125 A CN1295125 A CN 1295125A
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Abstract
The method includes the operating steps of preparation, packing and disinfection of medium for strain culture; small slant inoculation; slant culture first in constant temperature condition and then in room temperature for 3-10 days; wrapping cultured slant strain first with steam disinfected paper and then with steam disinfected polypropylene film; and preserving at room temperature or at 0-10 deg.c in a refrigerator. The present invention has low cost and needs no special equipment.
Description
The present invention relates to the microbial preservation method, specifically a kind of filamentous fungus passing, culturing, coating and preserving method.
In the prior art, the filamentous fungus under the pure culture state is divided into the formation spore and does not form two types in spore.The filamentous fungus of formation spore, its method for preserving also relatively extensively reach more than seven kinds, and do not form the filamentous fungus (basidiomycetes and sac fungi etc.) of spore, and its method for preserving just relatively limits to.Common reasonable method for preserving has: liquid nitrogen store method, the cultivation preservation method that goes down to posterity, white oil cover preservation method, wherein the liquid nitrogen preservation method often is subjected to equipment and the restriction of keeping funds, to such an extent as to general research and production unit do not have certain economic strength, unablely carry out this preservation work; The cultivation preservation method that goes down to posterity, though simple and easy to do, be fit to all kinds of bacterial classifications and all preservations of unit of culturing micro-organisms condition are arranged, it is insufficient to be, and retention period is short, need establish the special messenger and be responsible for inoculation regularly, and then consumption of raw material shortens with the preservation cycle and increases; White oil covers preservation method, is the covert cultivation store method that goes down to posterity, can the proper extension shelf time, do not need special device, expense is few, but the sterilization of mineral oil trouble, pollution in the operation is also unavoidable, moreover not all filamentous fungus all is fit to this method and carries out preservation, as mycorrhizal fungi during with this method preservation, the recovery situation is not fine just, surviving rate is extremely low, and therefore, this method can not be used widely.
In order to overcome above-mentioned deficiency, it is low to the purpose of this invention is to provide a kind of cost, does not need to set up specific installation, starting material wide material sources, good, the widely used filamentous fungus passing, culturing, coating and preserving method of simple and easy to do again, resuscitation effect.
To achieve these goals, technical scheme of the present invention is: the branch loading amount of cultivating strain substrate is abundant and the inclined-plane is little, puts, wraps up preservation then, specifically operation according to the following steps:
Handle early stage: cultivate the matrix preparation of bacterial classification according to a conventional method, quantitatively packing, steam sterilizing; Described quantitative minute loading amount is 20 * 2cm test tube, 15～18ml, 18 * 1.8cm test tube, 12～15ml, test tube 15 * 1.5cm test tube 5～10ml;
System inclined-plane inoculation: make little inclined-plane, then inoculation according to a conventional method; The gradient scope on described little inclined-plane: 20 * 20cm test tube is 2～6cm, and 18 * 1.8cm test tube is 2～4cm, and 15 * 1.5cm test tube is 2～3cm.
Temperature control: the slant strains that connects is carried out constant temperature culture according to a conventional method, cultured slant strains is displaced to room temperature placed 3～10 days;
Packing: behind paper, polypropylene film steam sterilizing, wrap cultured slant strains with paper bag earlier, tighten with polypropylene film Bao Yan again;
Preservation: it is to preserve in the refrigerator of 0＞T≤10 ℃ that the bacterial classification that bandages is placed room temperature or temperature T.
The present invention has following advantage:
1. cost is low.The present invention need not set up any new equipment, and the preserved material of cultured bacterial classification such as coated papers, can purchase under situation with good conditionsi, otherwise can collect from daily life neutralizes the waste reclaimation of other work system; In addition, inoculation time is long at interval, with the present invention 2 years inoculation once with inoculate the half a year of routine once than, its economic benefit is original 3 times.
2. simple and easy to do, high efficiency.The present invention is simple to operate; Do not need specific culture; Not only save human and material resources, also reduced passage number, to reach the purpose that does not change the bacterial classification proterties as far as possible, the bacterial classification shelf time is by extending to more than 1.5 years or 2 years original half a year.
3. resuscitation effect is good, applied range.Through the bacterial classification of preservation of the present invention, its recovery of switching back and growing state with recover with the bacterial classification of conventional method for preserving preservation in the past and growing state consistent, the present invention can be applicable to the preservation of all microbial strainss, particularly filamentous fungus.
Below in conjunction with embodiment the present invention is described in further detail.
It is abundant and the inclined-plane is little that the present invention cultivates the branch loading amount of strain substrate, puts, wraps up preservation then, specifically operation according to the following steps:
1. handle early stage: cultivate the matrix preparation of bacterial classification according to a conventional method, quantitatively packing, steam sterilizing; Described quantitative minute loading amount is 20 * 2cm test tube, 15～18ml, 18 * 1.8cm test tube, 12～15ml, test tube 15 * 1.5cm test tube 5～10ml;
2. system inclined-plane inoculation: make little inclined-plane, then inoculation according to a conventional method; The gradient scope on described little inclined-plane: 20 * 2cm test tube is 2～6cm, and 18 * 1.8cm test tube is 2～4cm, and 15 * 1.5cm test tube is 2～3cm
3. temperature control: the slant strains that connects is carried out constant temperature culture according to a conventional method, cultured slant strains is displaced to room temperature placed 3～10 days;
4. pack: behind paper, polypropylene film steam sterilizing, wrap cultured slant strains with paper bag earlier, tighten with polypropylene film Bao Yan again;
5. preservation: it is to preserve in the refrigerator of 0＞T≤10 ℃ that the bacterial classification that bandages is placed room temperature or temperature T.
Load onto in quantitative the branch, if the amount dress is many, opportunities for contamination is just many, if lacking of amount dress, it is fast that matrix is done, so quantitatively packing is a very important problem, the situation of about amount packing is used in inoculations in these per 3,4 months different from the past, with 18 * 1.8cm test tube is example, and the branch loading amount is adjusted to 15ml from 10ml;
Make about the inclined-plane, grasping with little inclined-plane is principle, and the too big mycelial surface-area in inclined-plane is just big, and that does is fast, especially on top; But the inclined-plane can not be too little, and too little meeting restriction mycelial growth is so inclined-plane gradient scope is adjusted to 2～4cm from 5～6cm (is example with 18 * 1.8 test tubes);
In temperature control, a kind of is room temperature preservation, another kind is to preserve in the refrigerator, the latter belongs to cryopreservation, and the present invention is limited in 0＞T≤10 ℃ to the temperature T of cryopreservation, is because approach room temperature greater than 10 ℃, do not belong to cryopreservation, during smaller or equal to 0 ℃, mycelium is easily endured cold, and resuscitation effect is bad.
Embodiment 1
The present invention operates according to the following steps:
Handle early stage: cultivate the matrix preparation of bacterial classification according to a conventional method, 20 * 2cm test tube is adopted in the fixation packing, and the branch loading amount is 17ml, steam sterilizing;
The inoculation of system inclined-plane: make little inclined-plane, the gradient on described little inclined-plane is 4cm, then inoculation according to a conventional method;
Intensity control: with the slant strains that connects routinely 25 ℃ of methods carry out constant temperature culture, cultured slant strains is displaced to room temperature placed 7 days;
Packing: behind kraft paper, polypropylene film steam sterilizing, wrap up tight cultured slant strains with kraft paper earlier, tighten with polypropylene film Bao Yan again;
Preservation: the bacterial classification that bandages is placed room temperature preservation.
Empirical tests: from the bacterial classification of 1997.12～1999.10 months preservation, its recovery of switching back and growing state with recover with the bacterial classification of conventional method for preserving preservation in the past and growing state consistent, its preservation period reaches 1 year 0 10 months.
Embodiment 2
The present invention operates according to the following steps:
Handle early stage: cultivate the matrix preparation of bacterial classification according to a conventional method, 18 * 1.8cm test tube is adopted in quantitatively packing, and the branch loading amount is 14ml, steam sterilizing;
The inoculation of system inclined-plane: make little inclined-plane, the gradient on described little inclined-plane is 3cm, then inoculation according to a conventional method;
Temperature control: with the slant strains that connects routinely 26 ℃ of methods carry out constant temperature culture, cultured slant strains is displaced to room temperature placed 10 days;
Packing: behind common blank sheet of paper, polypropylene film steam sterilizing, wrap cultured slant strains with the common white paper bag earlier, tighten with polypropylene film Bao Yan again;
Preservation: it is to preserve in 5 ℃ the refrigerator that the bacterial classification that bandages is placed temperature.
Empirical tests: from the bacterial classification of 1997.5～1999.10 months preservation, its recovery of switching back and growing state with recover with the bacterial classification of conventional method for preserving preservation in the past and growing state consistent, its preservation period reaches 2 years 05 months.
Embodiment 3
The present invention operates according to the following steps:
Handle early stage: cultivate the matrix preparation of bacterial classification according to a conventional method, 20 * 2cm test tube is adopted in quantitatively packing, and packing is changed to 15ml, steam sterilizing;
The inoculation of system inclined-plane: make little inclined-plane, the gradient on described little inclined-plane is 3cm, then inoculation according to a conventional method;
Temperature control: with the slant strains that connects routinely 28 ℃ of methods carry out constant temperature culture, cultured slant strains is displaced to room temperature placed 10 days;
Packing: behind kraft paper, polypropylene film steam sterilizing, wrap up tight cultured slant strains with kraft paper earlier, tighten with polypropylene film Bao Yan again;
Preservation: the bacterial classification that bandages is placed room temperature preservation.
Empirical tests: from the bacterial classification of 1997.7～1999.2 months preservation, its recovery of switching back and growing state with recover with the bacterial classification of conventional method for preserving preservation in the past and growing state consistent, its preservation period these 1 year 06 months.
Embodiment 4
The present invention operates according to the following steps:
Handle early stage: cultivate the matrix preparation of bacterial classification according to a conventional method, 18 * 1.8cm test tube is adopted in quantitatively packing, and the branch loading amount is 12ml, steam sterilizing;
The inoculation of system inclined-plane: make little inclined-plane, the gradient on described little inclined-plane is 4cm, then inoculation according to a conventional method;
Temperature control: with the slant strains that connects routinely 27 ℃ of methods carry out constant temperature culture, cultured slant strains is displaced to room temperature placed 4 days;
Packing: behind template, polypropylene film steam sterilizing, wrap up tight cultured slant strains with template earlier, tighten with polypropylene film Bao Yan again;
Preservation: it is to preserve in 2 ℃ the refrigerator that the bacterial classification that bandages is placed temperature.
Empirical tests: from the bacterial classification of 1997.5～1999.10 months preservation, its recovery of switching back and growing state with recover with the bacterial classification of conventional method for preserving preservation in the past and growing state consistent, its preservation period reaches 2 years 05 months.
The present invention also is applicable to 15 * 1.5cm test tube.
Claims (1)
1. filamentous fungus passing, culturing, coating and preserving method is characterized in that: the branch loading amount of cultivating strain substrate is abundant and the inclined-plane is little, puts, wraps up preservation then, specifically operation according to the following steps:
Handle early stage: cultivate the matrix preparation of bacterial classification according to a conventional method, quantitatively packing, steam sterilizing; Described quantitative minute loading amount is 20 * 2cm test tube, 15～18ml, 18 * 1.8cm test tube, 12～15ml, test tube 15 * 1.5cm test tube 5～10ml.
System inclined-plane inoculation: make little inclined-plane, then inoculation according to a conventional method; The gradient scope on described little inclined-plane: 20 * 2cm test tube is 2～6cm, and 18 * 1.8cm test tube is 2～4cm, and 15 * 1.5cm test tube is 2～3cm.
Temperature control: the slant strains that connects is carried out constant temperature culture according to a conventional method, cultured slant strains is displaced to room temperature placed 3～10 days;
Packing: behind paper, polypropylene film steam sterilizing, wrap cultured slant strains with paper bag earlier, tighten with polypropylene film Bao Yan again;
Preservation: it is to preserve in the refrigerator of 0＞T≤10 ℃ that the bacterial classification that bandages is placed room temperature or temperature T.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 99113397 CN1295125A (en) | 1999-11-03 | 1999-11-03 | Filamentous fungus passing, culturing, coating and preserving method |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 99113397 CN1295125A (en) | 1999-11-03 | 1999-11-03 | Filamentous fungus passing, culturing, coating and preserving method |
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| CN1295125A true CN1295125A (en) | 2001-05-16 |
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| CN 99113397 Pending CN1295125A (en) | 1999-11-03 | 1999-11-03 | Filamentous fungus passing, culturing, coating and preserving method |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1317381C (en) * | 2004-12-23 | 2007-05-23 | 李世杰 | Polysaccharose film of brevibacterium and its use in bacteria preservisation |
| CN102146343A (en) * | 2010-12-31 | 2011-08-10 | 吴海燕 | Storage method capable of keeping activity of nematode-trapping fungi for long time |
-
1999
- 1999-11-03 CN CN 99113397 patent/CN1295125A/en active Pending
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1317381C (en) * | 2004-12-23 | 2007-05-23 | 李世杰 | Polysaccharose film of brevibacterium and its use in bacteria preservisation |
| CN102146343A (en) * | 2010-12-31 | 2011-08-10 | 吴海燕 | Storage method capable of keeping activity of nematode-trapping fungi for long time |
| CN102146343B (en) * | 2010-12-31 | 2012-07-25 | 吴海燕 | Storage method capable of keeping activity of nematode-trapping fungi for long time |
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