CN102357423B - Potassium chloride flotation method of plant and soil nematodes - Google Patents
Potassium chloride flotation method of plant and soil nematodes Download PDFInfo
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- CN102357423B CN102357423B CN 201110267841 CN201110267841A CN102357423B CN 102357423 B CN102357423 B CN 102357423B CN 201110267841 CN201110267841 CN 201110267841 CN 201110267841 A CN201110267841 A CN 201110267841A CN 102357423 B CN102357423 B CN 102357423B
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Abstract
A potassium chloride flotation method of plant or soil nematodes relates to a flotation method of the plant or soil nematodes, and solves a problem that the existing cane sugar centrifugation floatation of the plant or soil nematodes is complicated in procedures, long in time using and, large in working amount, and consumes water and energy. Potassium chloride solution is added in a plant or soil sample, mixed, and stood for obtaining liquid supernatant and sediment, the sediment instead of the plant or soil sample to be repeatedly carried out the operation, and the obtained liquid supernatant is merged and then injected in a mesh screen backwashed by water. The potassium chloride flotation method uses high concentration potassium chloride solution to select the nematodes in flotation mode, complicated centrifugation procedures and heavy working amount are avoided, water using and centrifugation power using for water-washing the plant or soil sample can be greatly saved, and separation time is shortened by 1.5-6 hours. Compared with the traditional cane sugar separation method, the working amount reduces by 4/5, working time reduces by 70%-90%, working cost lowers by 2/3, and accuracy of a separation result is improved while the separation efficiency is guaranteed.
Description
Technical field
The present invention relates to the method for floating of a Plants or soil nematodes, be specifically related to potassium chloride flotation method.
Background technology
Nematode is the animal monoid of quantity maximum on the earth, is divided into Plant nematode, animal nematode etc. according to its somatomedin.Be widely studied at biology, geography and environmental science.It is the basic link of nematode research that nematode in the ecosystem is separated, and also is most important link.Obtaining the capacity nematode is the basic guarantee of studying.At present, the separation method of soil nematodes mainly adopts sucrose Centrifugal Flotation method, modified Baermann funnel method and tray method.
Modified Baermann funnel method is used for separating of worm alive only with the tray method, and separative efficiency is lower than sucrose Centrifugal Flotation method.Sucrose Centrifugal Flotation method is to stir evenly rear centrifugally with containing the plant of nematode or soil sample water, will precipitate with sucrose solution and stir evenly, centrifugal, precipitation again, the flushing of sieving; This method can be separated the long preservation sample that contains dead nematode, is not subjected to sampling time and operation to be subjected to the restriction of room temperature height, is that nematode separates method the most commonly used.But sucrose Centrifugal Flotation method complex steps, the time spent is longer, and workload is large, the water consumption power consumption.As the basis of nematode research, the method for obtaining the capacity nematode convenient, efficient and time-saving energy-saving remains to be probed into.
Summary of the invention
The objective of the invention is in order to solve the sucrose Centrifugal Flotation method complex steps of existing plant or soil nematodes, the time spent is longer, and workload is large, and the problem of water consumption power consumption the invention provides the potassium chloride flotation method of a Plants or soil nematodes.
The potassium chloride flotation method of plant of the present invention or soil nematodes is realized by following steps: one, take by weighing plant or pedotheque, then add in the Klorvess Liquid that concentration is 5mol/L and stir 5～20min, get suspension, wherein, the mass ratio of Klorvess Liquid and plant or pedotheque is 5～10 ﹕ 1; Two, the suspension that step 1 is obtained staticly settles 30～60min, gets supernatant and sediment; Three, the sediment that step 2 obtained replaces plant or the pedotheque in the step 1, by the method repetitive operation of step 1 and step 2 2～5 times, then discards sediment, obtains 3～6 portions of supernatants; Four, 3～6 portions of supernatants that step 3 obtained merge, and then inject 80 orders and the 500 purpose mesh screens that are nested together, and water backwash 500 order mesh screens get nematode again, finish the potassium chloride flotation method of plant or soil nematodes.
As long as avoid supernatant to spill when in the step 3 of the present invention the supernatant that merges being injected mesh screen.
The invention has the beneficial effects as follows: the present invention has directly adopted high concentration chlorination potassium solution (5mol/L) flotation nematode, avoided loaded down with trivial details centrifugal program and hard work amount, greatly saved water and the centrifugal electricity consumption of the washing of plant or pedotheque, and disengaging time has been shortened to 1.5 hours～6 hours; Solved in the sucrose Centrifugal Flotation method nematode damage problem centrifugal and twice precipitation produces twice; The sample that volume is different from water content carries out the saline solution of same ratio to be processed, and has reduced disturbing factor, guarantees the systematic error of nematode sample collection, has also improved the accuracy of separating resulting when guaranteeing separative efficiency.
Potassium chloride flotation method of the present invention is compared with traditional sucrose separation method, and workload has reduced 4/5, and the working time has reduced 70%～90%, and job costs have reduced by 2/3.Show the separative efficiency no significant difference of this method and sucrose Centrifugal Flotation method through the statistical check to test result.The present invention uses in research work, and the actual detection by carrying out in the research to state natural sciences fund " Caenorhabditis elegans HSPs is to the response mechanism of heavy metal stress " has obtained preferably technique effect.
The specific embodiment
Technical solution of the present invention is not limited to the following cited specific embodiment, also comprises any combination between each specific embodiment.
The specific embodiment one: present embodiment is that the potassium chloride flotation method of plant or soil nematodes is realized by following steps: one, take by weighing plant or pedotheque, then add in the Klorvess Liquid that concentration is 5mol/L and stir 5～20min, get suspension, wherein, the mass ratio of Klorvess Liquid and plant or pedotheque is 5～10 ﹕ 1; Two, the suspension that step 1 is obtained staticly settles 30～60min, gets supernatant and sediment; Three, the sediment that step 2 obtained replaces plant or the pedotheque in the step 1, by the method repetitive operation of step 1 and step 2 2～5 times, then discards sediment, obtains 3～6 portions of supernatants; Four, 3～6 portions of supernatants that step 3 obtained merge, and then inject 80 orders and the 500 purpose mesh screens that are nested together, and water backwash 500 order mesh screens get nematode again, finish the potassium chloride flotation method of plant or soil nematodes.
The beneficial effect of present embodiment is: the present invention has directly adopted high concentration chlorination potassium solution (5mol/L) flotation nematode, avoided loaded down with trivial details centrifugal program and hard work amount, greatly saved water and the centrifugal electricity consumption of the washing of plant or pedotheque, and disengaging time was shortened to 2 hours by 2 days; Solved in the sucrose Centrifugal Flotation method nematode damage problem centrifugal and twice precipitation produces twice; The sample that volume is different from water content carries out the saline solution of same ratio to be processed, and has reduced disturbing factor, guarantees the systematic error of nematode sample collection, has also improved the accuracy of separating resulting when guaranteeing separative efficiency.
In order to guarantee the working concentration of potassium chloride, in the time of the plant in the step 1 or pedotheque moisture content large (〉=80%), the mass ratio of Klorvess Liquid and plant or pedotheque is higher just passable, is 8～10 ﹕ 1 in the present embodiment; Moisture content is less than 80% the time in plant or the pedotheque, the mass ratio of Klorvess Liquid and plant or pedotheque lower just can, be 5～7 ﹕ 1.
As long as avoid supernatant to spill when in the present embodiment step 3 supernatant that merges being injected mesh screen.
In order to verify the effect of present embodiment, carry out following test: one, take by weighing 50g pedotheque (picking up from Changling Aneurolepidium Chinense Meadow soil), then add in the Klorvess Liquid that concentration is 5mol/L and stir 20min, get suspension, wherein, the mass ratio of Klorvess Liquid and pedotheque is 10 ﹕ 1; Two, the suspension that step 1 is obtained staticly settles 60min, gets supernatant and sediment; Three, the sediment that step 2 obtained replaces the 50g pedotheque in the step 1, by the method repetitive operation of step 1 and step 24 times, then discards sediment, obtains 5 portions of supernatants; Four, 5 portions of supernatants that step 3 obtained merge, and then inject 80 orders and the 500 purpose mesh screens that are nested together, and water backwash 500 order mesh screens get 3287 nematodes again, finish the potassium chloride flotation method of earth nematode.Altogether used 100min just to finish the potassium chloride flotation of soil nematodes among the embodiment.
Carry out simultaneously following contrast test: adopt traditional sucrose separation method, to the 50g pedotheque (with take among the embodiment with vicinal soil, pick up from Changling Aneurolepidium Chinense Meadow soil) carry out flotation, concrete sucrose separation method is: the 50g pedotheque is added about 500mL water fully stir evenly, behind the static 5min, respectively successively by 60,400 eye mesh screens, said process repeats 3 times, with clear water rush in the 400 purpose screen clothes material to beaker behind 20 ℃ of quiescent setting 12h, abandoning supernatant adds water stirring sediment and is placed in the centrifuge with 2000rmin
-1Centrifugal 5 minutes, abandon supernatant, adding concentration is 800gL
-1Sucrose solution stirs evenly, again with 2000rmin
-1Centrifugal 5min pours supernatant in 500 eye mesh screens water flushing, obtains 3355 nematodes.More than 20 hours have been spent altogether (2 days).
As seen, the potassium chloride flotation method of test is compared with traditional sucrose separation method, the separative efficiency no significant difference, and workload has reduced 4/5, and the working time was reduced to 100min by more than 20 hours, had reduced by about 90% working time, and job costs have reduced by 2/3.
The specific embodiment two: what present embodiment and the specific embodiment one were different is to add in the step 1 in the Klorvess Liquid that concentration is 5mol/L to stir 8～15min.Other step and parameter are identical with the specific embodiment one.
The specific embodiment three: what present embodiment and the specific embodiment one were different is to add in the step 1 in the Klorvess Liquid that concentration is 5mol/L to stir 10min.Other step and parameter are identical with the specific embodiment one.
The specific embodiment four: present embodiment and the specific embodiment one, two or three different be that the mass ratio of Klorvess Liquid and plant or pedotheque is 8 ﹕ 1 in the step 1.Other step and parameter and the specific embodiment one, two or three identical.
The specific embodiment five: what present embodiment was different from one of specific embodiment one to four is that the suspension that in the step 2 step 1 is obtained staticly settles 50min.Other step and parameter are identical with one of specific embodiment one to four.
Claims (5)
1. the potassium chloride flotation method of a Plants or soil nematodes, the potassium chloride flotation method that it is characterized in that plant or soil nematodes is realized by following steps: one, take by weighing plant or pedotheque, then add in the Klorvess Liquid that concentration is 5mol/L and stir 5～20min, get suspension, wherein, the mass ratio of Klorvess Liquid and plant or pedotheque is 5～10 ﹕ 1; Two, the suspension that step 1 is obtained staticly settles 30～60min, gets supernatant and sediment; Three, the sediment that step 2 obtained replaces plant or the pedotheque in the step 1, by the method repetitive operation of step 1 and step 2 2～5 times, then discards sediment, obtains 3～6 portions of supernatants; Four, 3～6 portions of supernatants that step 3 obtained merge, and then inject 80 orders and the 500 purpose mesh screens that are nested together, and water backwash 500 order mesh screens get nematode again, finish the potassium chloride flotation method of plant or soil nematodes.
2. the potassium chloride flotation method of a Plants according to claim 1 or soil nematodes is characterized in that adding in the step 1 in the Klorvess Liquid that concentration is 5mol/L and stirs 8～15min.
3. the potassium chloride flotation method of a Plants according to claim 1 or soil nematodes is characterized in that adding in the step 1 in the Klorvess Liquid that concentration is 5mol/L and stirs 10min.
4. according to claim 1, the potassium chloride flotation method of 2 or 3 described Plants or soil nematodes, it is characterized in that the mass ratio of Klorvess Liquid and plant or pedotheque is 8 ﹕ 1 in the step 1.
5. according to claim 1, the potassium chloride flotation method of 2 or 3 described Plants or soil nematodes, it is characterized in that the suspension that in the step 2 step 1 is obtained staticly settles 50min.
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| CN102607928A (en) * | 2012-03-20 | 2012-07-25 | 中国科学院东北地理与农业生态研究所 | Method for utilizing NaCl solution to separate nematode |
| CN107367439A (en) * | 2017-09-18 | 2017-11-21 | 任锦帅 | The assay method of detritus in a kind of bed mud |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101322500A (en) * | 2008-07-29 | 2008-12-17 | 董长江 | Environmental protection type soil nematode inhibitor |
| CN101398348A (en) * | 2008-11-03 | 2009-04-01 | 厦门大学 | Sampling apparatus for plant and nematode and detection method thereof |
| CN101849534A (en) * | 2010-06-22 | 2010-10-06 | 中国科学院东北地理与农业生态研究所 | Separation and detection device of entomopathogenic nematode, separation method of entomopathogenic nematode in soil and measuring method of population density |
| CN102084849A (en) * | 2010-11-19 | 2011-06-08 | 南京林业大学 | Method for preparing bacteria-free pine wood nematodes |
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Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101322500A (en) * | 2008-07-29 | 2008-12-17 | 董长江 | Environmental protection type soil nematode inhibitor |
| CN101398348A (en) * | 2008-11-03 | 2009-04-01 | 厦门大学 | Sampling apparatus for plant and nematode and detection method thereof |
| CN101849534A (en) * | 2010-06-22 | 2010-10-06 | 中国科学院东北地理与农业生态研究所 | Separation and detection device of entomopathogenic nematode, separation method of entomopathogenic nematode in soil and measuring method of population density |
| CN102084849A (en) * | 2010-11-19 | 2011-06-08 | 南京林业大学 | Method for preparing bacteria-free pine wood nematodes |
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