CN103875605A - Method for culturing Plectus bacterial-feeding nematodes - Google Patents
Method for culturing Plectus bacterial-feeding nematodes Download PDFInfo
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- CN103875605A CN103875605A CN201410089794.3A CN201410089794A CN103875605A CN 103875605 A CN103875605 A CN 103875605A CN 201410089794 A CN201410089794 A CN 201410089794A CN 103875605 A CN103875605 A CN 103875605A
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Abstract
The invention relates to a method for culturing Plectus bacterial-feeding nematodes. According to the method, the Plectus bacterial-feeding nematodes can be controlled indoors and sustainably cultured. The method includes the steps of (1), NaCl, KCl, yeast powder, peptone, tryptone and agar are weighed, taken and placed in a beaker, the constant volume is carried out on the mixture through distilled water, then the mixture is sterilized, and culture glue is obtained after cooling; (2), after being mashed, the culture glue is added to sterile pure water and stirred evenly, KH2PO4, K2HPO4, CaCl2, MgSO4 and cholesterol are added, then the pH value is adjusted through phosphoric acid, and a culture solution is obtained; (3), escherichia coli is added to the culture solution, so that bacterial liquid is obtained through culturing; (4), the bacterial liquid is taken and poured into a culture dish, the Plectus bacterial-feeding nematodes are added to the culture dish, and culturing is finished. The method achieves controllable culture of the bacterial-feeding Plectus nematodes and is applied to the microbial field.
Description
Technical field
The present invention relates to a kind of cultural method of eating bacterium class Plectus nematode.
Background technology
Nematode is described as the animal monoid of most abundant on the earth, is the indicator organism that carries out natural, ecological health risk assessment and environmental pollution evaluation.Realize this ecological monitoring that utilizes nematode to pollute to define, need to, by indoor mode and field accusation test, disclose the stress response mechanism of nematode under contaminated environment; Indoor, nematode being carried out to controlled cultivation and realize, is basis and the guarantee of utility nematode as indicator organism.It is the soil nematodes that important ecological functions are enriched, had to a class quantity that coiling belongs to (Plectus) food bacterium nematode, and life medium need to have attachment to like and move about again, high to the moisture requirement of living environment.
Plectus nematode cannot long term survival on conventional nematode culture medium (NGM medium), and can not grow in liquid medium within.Cannot controlledly cultivating of food bacterium class Plectus nematode at present, impact utilizes food bacterium class Plectus nematode to carry out the carrying out of constructed experiment.
Summary of the invention
The object of the invention is the problem in order to solve the indoor controlled sustainable cultivation of food bacterium class Plectus nematode, and the food bacterium class Plectus cultural method of nematode is provided.
The cultural method that the present invention eats bacterium class Plectus nematode carries out according to the following steps: one, take 1.5～2.5g NaCl, 1.0～2.0g KCl, 0.4～0.6g dusty yeast, 0.8～1.2g peptone, 0.8～1.2g tryptone and 0.8～1.2g agar, put into beaker, then be settled to 100mL with distilled water, put into again 120 DEG C of sterilizing 20min of high-pressure steam sterilizing pan, obtain cultivating glue after cooling; Two, cultivation glue step 1 being obtained adds in 100ml sterile water after smashing to pieces, stirs, and adds 20～50mgKH
2pO
4, 20～50mg K
2hPO
4, 10mg CaCl
2, 10mg MgSO
4with 10mg cholesterol, then the phosphorus acid for adjusting pH value that is 98% by mass concentration is 6.0, obtains culture fluid; Three, in the culture fluid obtaining in step 2, adding 0.1～0.2mL concentration is 1 × 10
9individual/mL～2 × 10
9the Escherichia coli of individual/mL are cultivated 4～6h in 37 DEG C of incubators, obtain bacterium liquid; Four, get the bacterium liquid 20ml that step 3 obtains and pour in culture dish, adding coiling to belong to food bacterium nematode, put into 20 DEG C of constant incubators and cultivate, complete.
The present invention has realized the indoor continual and steady cultivation to food bacterium class Plectus nematode.
Beneficial effect of the present invention:
Coiling belongs to (Plectus) food bacterium nematode does not grow on conventional NGM medium, in the culture fluid of fully liquid, does not also grow.The invention solves this class and like a difficult problem for the indoor lasting cultivation of nematode of moving about, this medium has solid-liquid mixed characteristic, simulate coiling and belonged to the habitat of (Plectus) food bacterium nematode in capillary soil water layer, met to greatest extent coiling and belonged to the requirement of (Plectus) food bacterium nematode to water threshold value.Utilize this solid-liquid mixed culture medium culture of bacteria, also improved the cover degree of bacterium in medium, ensured that this class nematode that moves about fully obtains food.The inventive method has realized the controlled sustainable cultivation of food bacterium class Plectus nematode, is conducive to eat bacterium class Plectus nematode and carries out the carrying out of constructed experiment.
Brief description of the drawings
Fig. 1 is the existence picture of food bacterium class Plectus nematode in test 1 medium;
Fig. 2 is the existence picture of food bacterium class Plectus nematode in test 1 medium.
Embodiment
Embodiment one: the cultural method of present embodiment food bacterium class Plectus nematode carries out according to the following steps: one, take 1.5～2.5g NaCl, 1.0～2.0g KCl, 0.4～0.6g dusty yeast, 0.8～1.2g peptone, 0.8～1.2g tryptone and 0.8～1.2g agar, put into beaker, then be settled to 100mL with distilled water, put into again 120 DEG C of sterilizing 20 min of high-pressure steam sterilizing pan, obtain cultivating glue after cooling; Two, cultivation glue step 1 being obtained adds in 100ml sterile water after smashing to pieces, stirs, and adds 20～50mgKH
2pO
4, 20～50mg K
2hPO
4, 10mg CaCl
2, 10mg MgSO
4with 10mg cholesterol, then the phosphorus acid for adjusting pH value that is 98% by mass concentration is 6.0, obtains culture fluid; Three, in the culture fluid obtaining in step 2, adding 0.1～0.2mL concentration is 1 × 10
9individual/mL～2 × 10
9the Escherichia coli of individual/mL are cultivated 4～6h in 37 DEG C of incubators, obtain bacterium liquid; Four, get the bacterium liquid 20ml that step 3 obtains and pour in culture dish, adding coiling to belong to food bacterium nematode, put into 20 DEG C of constant incubators and cultivate, complete.
Present embodiment has realized the indoor continual and steady cultivation to food bacterium class Plectus nematode.
The beneficial effect of present embodiment:
Coiling belongs to (Plectus) food bacterium nematode does not grow on conventional NGM medium, in the culture fluid of fully liquid, does not also grow.Present embodiment has solved the move about difficult problem of nematode indoor lasting cultivation of this class happiness, this medium has solid-liquid mixed characteristic, simulate coiling and belonged to the habitat of (Plectus) food bacterium nematode in capillary soil water layer, met to greatest extent coiling and belonged to the requirement of (Plectus) food bacterium nematode to water threshold value.Utilize this solid-liquid mixed culture medium culture of bacteria, also improved the cover degree of bacterium in medium, ensured that this class nematode that moves about fully obtains food.
Embodiment two: present embodiment is identical with embodiment two: the cultural method of food bacterium class Plectus nematode carries out according to the following steps: one, take 1.5～2.0g NaCl, 1.5～2.0g KCl, 0.4～0.6g dusty yeast, 0.8～1.2g peptone, 0.8～1.2g tryptone and 0.8～1.2g agar, put into beaker, then be settled to 100mL with distilled water, put into again 120 DEG C of sterilizing 20 min of high-pressure steam sterilizing pan, obtain cultivating glue after cooling; Two, cultivation glue step 1 being obtained adds in 100ml sterile water after smashing to pieces, stirs, and adds 30～40mgKH
2pO
4, 30～40mg K
2hPO
4, 10mg CaCl
2, 10mg MgSO
4with 10mg cholesterol, then the phosphorus acid for adjusting pH value that is 98% by mass concentration is 6.0, obtains culture fluid; Three, in the culture fluid obtaining in step 2, adding 0.1～0.2mL concentration is 1 × 10
9individual/mL～2 × 10
9the Escherichia coli of individual/mL are cultivated 4～6 hours in 37 DEG C of incubators; Four, get the bacterium liquid 20ml that step 3 obtains and pour in culture dish, adding coiling to belong to food bacterium nematode, put into 20 DEG C of constant incubators and cultivate, complete.
Other is identical with embodiment one.
Embodiment three: present embodiment is different from embodiment one or two: the cultural method of food bacterium class Plectus nematode carries out according to the following steps: one, take 1.5～2.5g NaCl, 1.0～2.0g KCl, 0.4～0.6g dusty yeast, 0.8～1.2g peptone, 0.8～1.2g tryptone and 0.8～1.2g agar, put into beaker, then be settled to 100mL with distilled water, put into again 120 DEG C of sterilizing 20 min of high-pressure steam sterilizing pan, obtain cultivating glue after cooling; Two, after cultivation glue step 1 being obtained is smashed to pieces, add in 100ml sterile water, after stirring, add 20～50mgKH
2pO
4, 20～50mg K
2hPO
4, 10mg CaCl
2, 10mg MgSO
4with 10mg cholesterol, then the phosphorus acid for adjusting pH value that is 98% by mass concentration is 6.0, obtains culture fluid; Three, in the culture fluid obtaining in step 2, adding 0.1～0.2mL concentration is 2 × 10
9the Escherichia coli of individual/mL are cultivated 5 hours in 37 DEG C of incubators; Four, get the bacterium liquid that 20ml step 3 obtains and pour in 9cm culture dish, adding coiling to belong to food bacterium nematode, put into 20 DEG C of constant incubators and cultivate, complete.
Other is identical with embodiment one or two.
Embodiment four: present embodiment is different from one of embodiment one to three: the cultural method of food bacterium class Plectus nematode carries out according to the following steps: one, take 2.5g NaCl, 2.0g KCl, 0.6g dusty yeast, 0.8g peptone, 0.8g tryptone and 0.8g agar, put into beaker, be settled to 100mL with distilled water, put into 120 DEG C of sterilizing 20 min of high-pressure steam sterilizing pan, obtain cultivating glue after cooling; Two, cultivation glue step 1 being obtained, after smashing to pieces, adds in 100ml sterile water with glass bar, after stirring, adds 20mgKH
2pO
4, 50mg K
2hPO
4, 10mg CaCl
2, 10mgMgSO
4with 10mg cholesterol, then the phosphorus acid for adjusting pH value that is 98% by mass concentration is 6.0, obtains culture fluid; Three, in the culture fluid obtaining in step 2, adding 0.1～0.2mL concentration is 1 × 10
9individual/mL～2 × 10
9the Escherichia coli of individual/mL are cultivated 4～6h in 37 DEG C of incubators, obtain bacterium liquid; Four, get the bacterium liquid that 20ml step 3 obtains and pour in 9cm culture dish, adding coiling to belong to food bacterium nematode, put into 20 DEG C of constant incubators and cultivate, complete.Other is identical with one of embodiment one to three.
Embodiment five: present embodiment is different from one of embodiment one to four: the cultural method of food bacterium class Plectus nematode carries out according to the following steps: one, take 2.5g NaCl, 2.0g KCl, 0.6g dusty yeast, 0.8g peptone, 0.8g tryptone and 0.8g agar, put into 250mL beaker, be settled to 100mL with distilled water, put into 120 DEG C of sterilizing 20 min of high-pressure steam sterilizing pan, obtain cultivating glue after cooling; Two, cultivation glue step 1 being obtained, after smashing to pieces, adds in 100ml sterile water with glass bar, after stirring, adds 20mgKH
2pO
4, 50mg K
2hPO
4, 10mg CaCl
2, 10mgMgSO
4with 10mg cholesterol, then the phosphorus acid for adjusting pH value that is 98% by mass concentration is 6.0, obtains culture fluid; Three, in the culture fluid obtaining in step 2, adding 0.1～0.2mL concentration is 12 × 10
9the Escherichia coli of individual/mL are cultivated 6h in 37 DEG C of incubators, obtain bacterium liquid; Four, get the bacterium liquid that 20ml step 3 obtains and pour in 9cm culture dish, adding coiling to belong to food bacterium nematode, put into 20 DEG C of constant incubators and cultivate, complete.Other is identical with one of embodiment one to four.
Embodiment six: present embodiment is different from one of embodiment one to five: the cultural method of food bacterium class Plectus nematode carries out according to the following steps: one, take 1.5g NaCl, 1.0g KCl, 0.4g dusty yeast, 0.8g peptone, 0.8g tryptone and 1.0g agar, put into 250mL beaker, be settled to 100mL with distilled water, put into 120 DEG C of sterilizing 20 min of high-pressure steam sterilizing pan, obtain cultivating glue after cooling; Two, cultivation glue step 1 being obtained, after smashing to pieces, adds in 100ml sterile water with glass bar, after stirring, adds 20mgKH
2pO
4, 20mg K
2hPO
4, 10mg CaCl
2, 10mgMgSO
4with 10mg cholesterol, then the phosphorus acid for adjusting pH value that is 98% by mass concentration is 6.0, obtains culture fluid; Three, in the culture fluid obtaining in step 2, adding 0.1～0.2mL concentration is 1 × 10
9the Escherichia coli of individual/mL/mL are cultivated 6 hours in 37 DEG C of incubators, obtain bacterium liquid; Four, get the bacterium liquid that 20ml step 3 obtains and pour in 9cm culture dish, adding coiling to belong to food bacterium nematode, put into 20 DEG C of constant incubators and cultivate, complete.Other is identical with one of embodiment one to five.
Embodiment seven: present embodiment is different from one of embodiment one to six: the cultural method of food bacterium class Plectus nematode carries out according to the following steps: one, take 2.0g NaCl, 1.5g KCl, 0.5g dusty yeast, 1.0g peptone, 1.0g tryptone and 1.0g agar, put into 250mL beaker, be settled to 100mL with distilled water, put into 120 DEG C of sterilizing 20 min of high-pressure steam sterilizing pan, obtain cultivating glue after cooling; Two, cultivation glue step 1 being obtained, after smashing to pieces, adds in 100ml sterile water with glass bar, after stirring, adds 25mgKH
2pO
4, 25mg K
2hPO
4, 10mg CaCl
2, 10mgMgSO
4with 10mg cholesterol, then the phosphorus acid for adjusting pH value that is 98% by mass concentration is 6.0, obtains culture fluid; Three, in the culture fluid obtaining in step 2, adding 0.1～0.2mL concentration is 1 × 10
9individual/mL～2 × 10
9the Escherichia coli of individual/mL are cultivated 4 hours in 37 DEG C of incubators, obtain bacterium liquid; Four, get the bacterium liquid that 20ml step 3 obtains and pour in 9cm culture dish, adding coiling to belong to food bacterium nematode, put into 20 DEG C of constant incubators and cultivate, complete.Other is identical with one of embodiment one to six.
Embodiment eight: present embodiment is different from one of embodiment one to seven: the cultural method of food bacterium class Plectus nematode carries out according to the following steps: one, take 2.5g NaCl, 2.0g KCl, 0.6g dusty yeast, 1.2g peptone, 1.2g tryptone and 1.2g agar, put into 250mL beaker, be settled to 100mL with distilled water, put into 120 DEG C of sterilizing 20 min of high-pressure steam sterilizing pan, obtain cultivating glue after cooling; Two, cultivation glue step 1 being obtained, after smashing to pieces, adds in 100ml sterile water with glass bar, after stirring, adds 50mgKH
2pO
4, 50mg K
2hPO
4, 10mg CaCl
2, 10mgMgSO
4with 10mg cholesterol, then the phosphorus acid for adjusting pH value that is 98% by mass concentration is 6.0, obtains culture fluid; Three, in the culture fluid obtaining in step 2, adding 0.1～0.2mL concentration is 2 × 10
9the Escherichia coli of individual/mL are cultivated 6 hours in 37 DEG C of incubators, obtain bacterium liquid; Four, get the bacterium liquid that 20ml step 3 obtains and pour in 9cm culture dish, adding coiling to belong to food bacterium nematode, put into 20 DEG C of constant incubators and cultivate, complete.Other is identical with one of embodiment one to seven.
By following verification experimental verification beneficial effect of the present invention:
The cultural method of test 1, this test food bacterium class Plectus nematode is as follows: one, take 2.5g NaCl, 2.0g KCl, 0.6g dusty yeast, 1.2g peptone, 1.2g tryptone and 1.2g agar, put into 250mL beaker, be settled to 100mL with distilled water, put into 120 DEG C of sterilizing 20 min of high-pressure steam sterilizing pan, obtain cultivating glue after cooling; Two, cultivation glue step 1 being obtained, after smashing to pieces, adds in 100ml sterile water with glass bar, after stirring, adds 50mgKH
2pO
4, 50mg K
2hPO
4, 10mg CaCl
2, 10mg MgSO
4with 10mg cholesterol, then the phosphorus acid for adjusting pH value that is 98% by mass concentration is 6.0, obtains culture fluid; Three, in the culture fluid obtaining in step 2, adding 0.1～0.2mL concentration is 2 × 10
9the Escherichia coli of individual/mL are cultivated 6 hours in 37 DEG C of incubators, obtain bacterium liquid; Four, get the bacterium liquid that 20ml step 3 obtains and pour in 9cm culture dish, adding coiling to belong to food bacterium nematode, put into 20 DEG C of constant incubators and cultivate 10d, complete.
Fig. 1 and Fig. 2 are the existence figure of food bacterium class Plectus nematode in this test medium, from Fig. 1 and Fig. 2, cultivate that within 10 days, to eat afterwards the existence of bacterium class Plectus nematode in this test medium good, prove that this test method has realized the indoor controlled sustainable cultivation of food bacterium class Plectus nematode.
Claims (8)
1. the cultural method of food bacterium class Plectus nematode, it is characterized in that the cultural method of eating bacterium class Plectus nematode carries out according to the following steps: one, take 1.5～2.5g NaCl, 1.0～2.0g KCl, 0.4～0.6g dusty yeast, 0.8～1.2g peptone, 0.8～1.2g tryptone and 0.8～1.2g agar, put into beaker, then be settled to 100mL with distilled water, put into again 120 DEG C of sterilizing 20 min of high-pressure steam sterilizing pan, obtain cultivating glue after cooling; Two, cultivation glue step 1 being obtained adds in 100ml sterile water after smashing to pieces, stirs, and adds 20～50mgKH
2pO
4, 20～50mg K
2hPO
4, 10mg CaCl
2, 10mg MgSO
4with 10mg cholesterol, then the phosphorus acid for adjusting pH value that is 98% by mass concentration is 6.0, obtains culture fluid; Three, in the culture fluid obtaining in step 2, adding 0.1～0.2mL concentration is 1 × 10
9individual/mL～2 × 10
9the Escherichia coli of individual/mL are cultivated 4～6h in 37 DEG C of incubators, obtain bacterium liquid; Four, get the bacterium liquid 20ml that step 3 obtains and pour in culture dish, adding coiling to belong to food bacterium nematode, put into 20 DEG C of constant incubators and cultivate, complete.
2. the cultural method of food bacterium class Plectus nematode according to claim 1, its characteristic is that the cultural method of eating bacterium class Plectus nematode carries out according to the following steps: one, take 1.5～2.0g NaCl, 1.5～2.0g KCl, 0.4～0.6g dusty yeast, 0.8～1.2g peptone, 0.8～1.2g tryptone and 0.8～1.2g agar, put into beaker, then be settled to 100mL with distilled water, put into again 120 DEG C of sterilizing 20 min of high-pressure steam sterilizing pan, obtain cultivating glue after cooling; Two, cultivation glue step 1 being obtained adds in 100ml sterile water after smashing to pieces, stirs, and adds 30～40mgKH
2pO
4, 30～40mg K
2hPO
4, 10mg CaCl
2, 10mg MgSO
4with 10mg cholesterol, then the phosphorus acid for adjusting pH value that is 98% by mass concentration is 6.0, obtains culture fluid; Three, in the culture fluid obtaining in step 2, adding 0.1～0.2mL concentration is 1 × 10
9individual/mL～2 × 10
9the Escherichia coli of individual/mL are cultivated 4～6 hours in 37 DEG C of incubators; Four, get the bacterium liquid 20ml that step 3 obtains and pour in culture dish, adding coiling to belong to food bacterium nematode, put into 20 DEG C of constant incubators and cultivate, complete.
3. the cultural method of food bacterium class Plectus nematode according to claim 1, its characteristic is that the cultural method of eating bacterium class Plectus nematode carries out according to the following steps: one, take 1.5～2.5g NaCl, 1.0～2.0g KCl, 0.4～0.6g dusty yeast, 0.8～1.2g peptone, 0.8～1.2g tryptone and 0.8～1.2g agar, put into beaker, then be settled to 100mL with distilled water, put into again 120 DEG C of sterilizing 20 min of high-pressure steam sterilizing pan, obtain cultivating glue after cooling; Two, after cultivation glue step 1 being obtained is smashed to pieces, add in 100ml sterile water, after stirring, add 20～50mgKH
2pO
4, 20～50mg K
2hPO
4, 10mg CaCl
2, 10mg MgSO
4with 10mg cholesterol, then the phosphorus acid for adjusting pH value that is 98% by mass concentration is 6.0, obtains culture fluid; Three, in the culture fluid obtaining in step 2, adding 0.1～0.2mL concentration is 2 × 10
9the Escherichia coli of individual/mL are cultivated 5 hours in 37 DEG C of incubators; Four, get the bacterium liquid that 20ml step 3 obtains and pour in 9cm culture dish, adding coiling to belong to food bacterium nematode, put into 20 DEG C of constant incubators and cultivate, complete.
4. the cultural method of food bacterium class Plectus nematode according to claim 1, its characteristic is that the cultural method of eating bacterium class Plectus nematode carries out according to the following steps: one, take 2.5g NaCl, 2.0g KCl, 0.6g dusty yeast, 0.8g peptone, 0.8g tryptone and 0.8g agar, put into 250mL beaker, be settled to 100mL with distilled water, put into 120 DEG C of sterilizing 20 min of high-pressure steam sterilizing pan, obtain cultivating glue after cooling; Two, cultivation glue step 1 being obtained, after smashing to pieces, adds in 100ml sterile water with glass bar, after stirring, adds 20mgKH
2pO
4, 50mg K
2hPO
4, 10mg CaCl
2, 10mg MgSO
4with 10mg cholesterol, then the phosphorus acid for adjusting pH value that is 98% by mass concentration is 6.0, obtains culture fluid; Three, in the culture fluid obtaining in step 2, adding 0.1～0.2mL concentration is 1 × 10
9individual/mL～2 × 10
9the Escherichia coli of individual/mL are cultivated 4～6h in 37 DEG C of incubators, obtain bacterium liquid; Four, get the bacterium liquid that 20ml step 3 obtains and pour in 9cm culture dish, adding coiling to belong to food bacterium nematode, put into 20 DEG C of constant incubators and cultivate, complete.
5. the cultural method of food bacterium class Plectus nematode according to claim 1, its characteristic is that the cultural method of eating bacterium class Plectus nematode carries out according to the following steps: one, take 2.5g NaCl, 2.0g KCl, 0.6g dusty yeast, 0.8g peptone, 0.8g tryptone and 0.8g agar, put into 250mL beaker, be settled to 100mL with distilled water, put into 120 DEG C of sterilizing 20 min of high-pressure steam sterilizing pan, obtain cultivating glue after cooling; Two, cultivation glue step 1 being obtained, after smashing to pieces, adds in 100ml sterile water with glass bar, after stirring, adds 20mgKH
2pO
4, 50mg K
2hPO
4, 10mg CaCl
2, 10mg MgSO
4with 10mg cholesterol, then the phosphorus acid for adjusting pH value that is 98% by mass concentration is 6.0, obtains culture fluid; Three, in the culture fluid obtaining in step 2, adding 0.1～0.2mL concentration is 12 × 10
9the Escherichia coli of individual/mL are cultivated 6h in 37 DEG C of incubators, obtain bacterium liquid; Four, get the bacterium liquid that 20ml step 3 obtains and pour in 9cm culture dish, adding coiling to belong to food bacterium nematode, put into 20 DEG C of constant incubators and cultivate, complete.
6. the cultural method of food bacterium class Plectus nematode according to claim 1, its characteristic is that the cultural method of eating bacterium class Plectus nematode carries out according to the following steps: one, take 1.5g NaCl, 1.0g KCl, 0.4g dusty yeast, 0.8g peptone, 0.8g tryptone and 1.0g agar, put into 250mL beaker, be settled to 100mL with distilled water, put into 120 DEG C of sterilizing 20 min of high-pressure steam sterilizing pan, obtain cultivating glue after cooling; Two, cultivation glue step 1 being obtained, after smashing to pieces, adds in 100ml sterile water with glass bar, after stirring, adds 20mgKH
2pO
4, 20mg K
2hPO
4, 10mg CaCl
2, 10mg MgSO
4with 10mg cholesterol, then the phosphorus acid for adjusting pH value that is 98% by mass concentration is 6.0, obtains culture fluid; Three, in the culture fluid obtaining in step 2, adding 0.1～0.2mL concentration is 1 × 10
9the Escherichia coli of individual/mL/mL are cultivated 6 hours in 37 DEG C of incubators, obtain bacterium liquid; Four, get the bacterium liquid that 20ml step 3 obtains and pour in 9cm culture dish, adding coiling to belong to food bacterium nematode, put into 20 DEG C of constant incubators and cultivate, complete.
7. the cultural method of food bacterium class Plectus nematode according to claim 1, its characteristic is that the cultural method of eating bacterium class Plectus nematode carries out according to the following steps: one, take 2.0g NaCl, 1.5g KCl, 0.5g dusty yeast, 1.0g peptone, 1.0g tryptone and 1.0g agar, put into 250mL beaker, be settled to 100mL with distilled water, put into 120 DEG C of sterilizing 20 min of high-pressure steam sterilizing pan, obtain cultivating glue after cooling; Two, cultivation glue step 1 being obtained, after smashing to pieces, adds in 100ml sterile water with glass bar, after stirring, adds 25mgKH
2pO
4, 25mg K
2hPO
4, 10mg CaCl
2, 10mg MgSO
4with 10mg cholesterol, then the phosphorus acid for adjusting pH value that is 98% by mass concentration is 6.0, obtains culture fluid; Three, in the culture fluid obtaining in step 2, adding 0.1～0.2mL concentration is 1 × 10
9individual/mL～2 × 10
9the Escherichia coli of individual/mL are cultivated 4 hours in 37 DEG C of incubators, obtain bacterium liquid; Four, get the bacterium liquid that 20ml step 3 obtains and pour in 9cm culture dish, adding coiling to belong to food bacterium nematode, put into 20 DEG C of constant incubators and cultivate, complete.
8. the cultural method of food bacterium class Plectus nematode according to claim 1, its characteristic is that the cultural method of eating bacterium class Plectus nematode carries out according to the following steps: one, take 2.5g NaCl, 2.0g KCl, 0.6g dusty yeast, 1.2g peptone, 1.2g tryptone and 1.2g agar, put into 250mL beaker, be settled to 100mL with distilled water, put into 120 DEG C of sterilizing 20 min of high-pressure steam sterilizing pan, obtain cultivating glue after cooling; Two, cultivation glue step 1 being obtained, after smashing to pieces, adds in 100ml sterile water with glass bar, after stirring, adds 50mgKH
2pO
4, 50mg K
2hPO
4, 10mg CaCl
2, 10mg MgSO
4with 10mg cholesterol, then the phosphorus acid for adjusting pH value that is 98% by mass concentration is 6.0, obtains culture fluid; Three, in the culture fluid obtaining in step 2, adding 0.1～0.2mL concentration is 2 × 10
9the Escherichia coli of individual/mL are cultivated 6 hours in 37 DEG C of incubators, obtain bacterium liquid; Four, get the bacterium liquid that 20ml step 3 obtains and pour in 9cm culture dish, adding coiling to belong to food bacterium nematode, put into 20 DEG C of constant incubators and cultivate, complete.
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN105891177A (en) * | 2016-04-13 | 2016-08-24 | 中国科学院东北地理与农业生态研究所 | Method for detecting trace BPA (bisphenol A) through nematode dyeing with Nile red |
| CN113317281A (en) * | 2021-06-11 | 2021-08-31 | 杭州师范大学 | Method for enrichment culture of marine edible bacterium nematodes |
| CN113396868A (en) * | 2021-06-29 | 2021-09-17 | 中国科学院南京土壤研究所 | In-situ enrichment method for nematodes in soil and application thereof |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2683799B2 (en) * | 1992-11-06 | 1997-12-03 | 王子製紙株式会社 | Insect parasitic nematode growth medium |
| RU2168893C1 (en) * | 2000-03-21 | 2001-06-20 | Всероссийский институт защиты растений | Method of entomopathogenic nematode biomass preparing |
-
2014
- 2014-03-12 CN CN201410089794.3A patent/CN103875605B/en not_active Expired - Fee Related
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2683799B2 (en) * | 1992-11-06 | 1997-12-03 | 王子製紙株式会社 | Insect parasitic nematode growth medium |
| RU2168893C1 (en) * | 2000-03-21 | 2001-06-20 | Всероссийский институт защиты растений | Method of entomopathogenic nematode biomass preparing |
Non-Patent Citations (6)
| Title |
|---|
| 冯志新: "《植物线虫学》", 30 May 2001, article "食细菌线虫的培养", pages: 197-198 * |
| 刘志典等: "土壤线虫培养方法研究", 《广东农业科学》, no. 5, 10 March 2013 (2013-03-10), pages 60 - 62 * |
| 李有全等: "秀丽隐杆线虫的培养与保存研究", 《中国兽医科学》, vol. 41, no. 10, 31 October 2011 (2011-10-31), pages 1001 - 1004 * |
| 李清源等: "秀丽隐杆线虫实验室培养方法的研究", 《中国畜牧兽医学会家畜寄生虫学分会第六次代表大会暨第十一次学术研讨会论文集》, 18 November 2011 (2011-11-18), pages 331 * |
| 李辉信等: "土壤食细菌线虫的分离和富集培养方法", 《南京农业大学学报》, vol. 25, no. 2, 30 April 2002 (2002-04-30), pages 71 - 74 * |
| 毛小芳等: "土壤食细菌线虫的原位富集培养方法", 《生态学报》, vol. 27, no. 2, 28 February 2007 (2007-02-28), pages 650 - 654 * |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105891177A (en) * | 2016-04-13 | 2016-08-24 | 中国科学院东北地理与农业生态研究所 | Method for detecting trace BPA (bisphenol A) through nematode dyeing with Nile red |
| CN105891177B (en) * | 2016-04-13 | 2019-02-19 | 中国科学院东北地理与农业生态研究所 | Nematode dyeing detection trace BPA method is carried out using Nile red |
| CN113317281A (en) * | 2021-06-11 | 2021-08-31 | 杭州师范大学 | Method for enrichment culture of marine edible bacterium nematodes |
| CN113396868A (en) * | 2021-06-29 | 2021-09-17 | 中国科学院南京土壤研究所 | In-situ enrichment method for nematodes in soil and application thereof |
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