CN103688908A - Enrichment culture method for aphelenchoidessp. - Google Patents
Enrichment culture method for aphelenchoidessp. Download PDFInfo
- Publication number
- CN103688908A CN103688908A CN201310696738.1A CN201310696738A CN103688908A CN 103688908 A CN103688908 A CN 103688908A CN 201310696738 A CN201310696738 A CN 201310696738A CN 103688908 A CN103688908 A CN 103688908A
- Authority
- CN
- China
- Prior art keywords
- nematode
- sword
- fungi
- culture dish
- food
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Landscapes
- Agricultural Chemicals And Associated Chemicals (AREA)
Abstract
The invention belongs to the biological technical field and particularly relates to an enrichment culture method for aphelenchoidessp.. The method comprises (1) aphelenchoidessp. separation; (2) aphelenchoidessp. identification; (3) aphelenchoidessp. surface sterilization; (4) aphelenchoidessp. enrichment culture. The method is simple and easy to implement, the survival rate of aphelenchoidessp. is high, the individual quality of aphelenchoidessp. is good, the hereditary change is small, management is convenient, lots of aphelenchoidessp. can be obtained effectively, and technical support is provided for studies on key roles of aphelenchoidessp. in the whole soil food chain and effects of the interaction between the aphelenchoidessp. and fungi in the plant growth.
Description
Technical field
The invention belongs to biological technical field, be specifically related to the enrichment culture method that the sliding sword of a kind of soil belongs to food fungi nematode.
Background technology
In soil, the nematode of perch has comparative advantage with free-living nematode, and soil free-living nematode to take food bacterium nematode and food fungi nematode be main (both are referred to as food Micronema), account for 80% of free-living nematode.A lot of research shows, eats Micronema by predation edaphon, accelerates microbial reproduction, and by the nutrient of metabolic secretion and releasing microbe institute fixing, and then affect soil ecological process and plant growth.But this effect meeting because of the kind of different nematodes, microorganism and plant and the C/N nutrition condition of soil matrix different.At present, about the interaction of soil food fungi nematode and fungi and less to the report of plant growth, the rare report especially of the correlative study in contaminated soil especially.Such research can be furtherd investigate feeding habit and the critical role of trophic behaviour in soil food chain and the circulation of whole soil nutrient thereof of soil food fungi nematode, and the soil of pure culture food fungi nematode is the basis of research.Many research show sliding sword belong to food fungi nematode (
aphelenchoidessp.) soil pollution is had to stronger patience, be extensively present in all kinds of contaminated soils, its further investigation is significant.But, rarely have at present the enrichment culture method that belongs to food fungi nematode about the especially sliding sword of soil food fungi nematode.
Summary of the invention
The object of the present invention is to provide the sliding sword of a kind of enrichment culture belong to food fungi nematode (
aphelenchoidessp.) method, the method is simple and easy to use, and the reproduction rate that sliding sword belongs to food fungi nematode is high, and the sliding sword that can be a large amount of pure cultures of efficient preparation belongs to food fungi nematode provides technical guarantee.
The sliding sword of a kind of enrichment culture provided by the invention belongs to the method for food fungi nematode, and concrete steps are as follows:
1, slide sword belong to food fungi nematode (
aphelenchoidessp.) separation: from the fresh soil of field acquisition.Adopt improved tray method separated nematode from soil.
2, slide sword belong to food fungi nematode (
aphelenchoidessp.) evaluation: liquid in beaker is proceeded to culture dish, then choose this genus nematode with capillary according to the feature of sliding sword genus food fungi nematode under anatomical lens, then it is carried out to identification of living body under light microscope.
3, slide sword belong to food fungi nematode (
aphelenchoidessp.) surface sterilization: nematode is proceeded in the centrifuge tube of sterilizing, with streptomycin sulphate and cycloheximide mixed liquor, nematode is carried out to surface sterilization;
4, slide sword belong to food fungi nematode (
aphelenchoidessp.) enrichment culture: by fungi Botrytis cinerea (
botrytis cinereapers.) be inoculated on potato dextrose agar, adopt sealed membrane (Parafilm) by culture dish sealing, to prevent the evaporation of medium moisture and to pollute.Then culture dish is inverted and is put in constant incubator, 20-25 ℃ of dark culturing after one week, sliding sword and belongs to nematode access and cover with in the culture dish of uniformity Botrytis cinerea when Botrytis cinerea amount reproduction.Adopt sealed membrane (Parafilm) by culture dish sealing, culture dish faces up and puts into sizeable sterilizing box, 20-25 ℃ of dark culturing, whether routine observation culture dish is subject to the pollution of other fungies or acarid, after several weeks, grey spore disappears substantially, nematode amount reproduction.Repeat above step and can obtain a large amount of sliding sword genus food fungi nematodes.Wherein, the formula of potato dextrose agar is: in every 1000 ml potato dextrose agars, and potato 190-220 g, glucose 18-20 g, agar 15-20 g, all the other are water, pH value nature.
Improved tray method equipment needed thereby described in above-mentioned steps (1) is simple, operation is row easily, specific as follows: 10 object stainless steel screen traies to be put into supporting tray, place two-layer face tissue on screen tray, then soil sample uniform spreading on paper, in tray, add water until submergence soil.Be placed in again under 25 ℃ of temperature, dark condition separated.After 48 h, with 400 mesh sieves, collect nematode in beaker.After standing over night, with vavuum pump, slowly draw supernatant liquid, avoid disturbance lower floor liquid as far as possible.Now, lower floor's liquid has been assembled a large amount of nematodes.
In above-mentioned steps (2), the main identification mark of sliding sword genus food fungi nematode is as follows: 1. heat kill after death, female worm polypide is until abdomen is curved, the whole polypide of the common afterbody ventrad of male worm hook is " J " shape (if live body is fubaritic, can be killed rear definite, for identification of living body provides reference), live body nematode is serpentine more; 2. the head contracting of conventionally slightly hanging, headstock osteomalacia, lancet is thinner, and base portion expands, and length is generally 10-12 μ m, is generally no more than 20 μ m; 3. esophagus presoma is cylindrical, middle oesophageal bulb prosperity, oval or square circular, and middle esophagus ball clack is flourishing, and rear esophageal gland leaf is flourishing, the back side of covering intestines; 4. porus excretorius is arranged in level place after oesophageal bulb; 5. female worm tail is conical, and tail end form is various: blunt circle or carefully round, finger-like or bifurcated or have an abdomen prominent, have or anury point prominent, the prominent form of tail point is various; 6. male worm tail be conical, to outside of belly hook; Copulatory spicules are rose thorn shape, and Ji Ding and base beak are conventionally flourishing, without copulatory bursa.
In above-mentioned steps (3), in described mixed liquor, the quality-volumetric concentration of streptomycin sulphate is 0.80-1.20 g.L
-1; The mass body volume concentrations of cycloheximide is 0.015-0.02 g.L
-1; The time of nematode being carried out to surface sterilization is 15-20 min.
In the present invention, the medium through sterilizing in step (4) be by medium in high-pressure sterilizing pot, at 121 ℃ of temperature, sterilizing 30 min obtain.The store method of the sliding sword genus food fungi nematode obtaining after its enrichment culture is as follows: adopt sealed membrane (Parafilm) by culture dish sealing, then culture dish is kept in 4 ℃ of refrigerators standby.Every 5-6 month, sliding sword is belonged to food fungi nematode and carry out subculture cultivation, overlong time is the scarcity of the evaporation of medium moisture and food resource, and nematode can be dead, and the time is too short is financial resources and labour's waste.Its specific operation process is, culture dish is taken out from 4 ℃ of refrigerators, choose the medium of the wired worm of a little block length, put on the potato dextrose agar that covers with uniformity Botrytis cinerea, to after culture dish sealing, put into sizeable sterilizing box, 20-25 ℃ of dark culturing, and whether routine observation culture dish be subject to the pollution of other fungies or acarid, after three weeks by culture dish as for preserving in 4 ℃ of refrigerators.
Beneficial effect of the present invention is: its method is simple, it is high that sliding sword belongs to food fungi nematode reproductive survival rate, nematode Individual Quality is good, inheritance changes little, convenient management, the sliding sword that can effectively obtain a large amount of pure cultures belongs to food fungi nematode, for eating trophic behaviour and the whole soil food chain of fungi nematode in research soil, provides technical support.In addition, sliding sword belongs to food fungi nematode and takes advantage in contaminated soil, can utilize its trophic behaviour to hypha,hyphae, the mineralising of accelerating stubbornness nutrient in mycelia or spore in contaminated soil discharges, thereby in agricultural production, can promote the growth of plant in contaminated soil, aspect the reparation of contaminated soil, can improve phytoremediation efficiency.
Embodiment
Below in conjunction with embodiment, the present invention is described in further detail.
Embodiment 1
1, slide the separation that sword belongs to food fungi nematode: from Institute of Botany (Zhong Shan botanical garden, Nanjing) nursery lot, gather fresh soil.Adopt improved tray method separated nematode from soil.Concrete grammar is described below.
10 object stainless steel screen traies are put into supporting tray, and on screen tray, place two-layer face tissue, then soil sample uniform spreading is sprinkling upon on paper, in tray, add water until submergence soil.Whole device is put in incubator, separated under 20～25 ℃ of temperature, dark condition.Due to the hydrotaxis of nematode, add the effect of himself gravity, nematode departs from successively soil and enters in water and (be applicable to the nematode that enlivens in separated soil sample) through face tissue hole.After 48-60 h, screen tray is taken out, pour the liquid in tray into 400 mesh sieve dishes, constantly water is collected nematode in beaker after rinsing screen tray.After beaker standing over night, with vavuum pump, slowly draw supernatant liquid, and avoid disturbance lower floor liquid as far as possible.Now, lower floor's liquid has been assembled a large amount of nematodes.
2, slide the evaluation that sword belongs to food fungi nematode
Rarely have at present food fungi nematode, especially sliding sword belongs to the report of food fungi nematode pure culture, and one of difficult point is also the evaluation of nematode, particularly in enrichment culture process, and need to be by nematode identification of living body; Authentication method is specific as follows: the culture dish that liquid in beaker is proceeded to diameter 5 cm.After standing 5 min, the morphological feature that belongs to nematode according to sliding sword is chosen this genus nematode with capillary under anatomical lens, then by its identification of living body under Olympus Optical microscope.
The main identification mark of identifying foundation when sliding sword belongs to food fungi nematode is as follows: 1. after death, female worm polypide is until abdomen is curved for heat kill, and the whole polypide of the common afterbody ventrad of male worm hook is " J " shape, and live body nematode is serpentine more; (if live body is fubaritic, can be killed to determine afterwards, for identification of living body provide with reference to), the 2. head contracting of conventionally slightly hanging, headstock osteomalacia, lancet is thinner, base portion expands, length is generally 10-12 μ m, is generally no more than 20 μ m; 3. esophagus presoma is cylindrical, middle oesophageal bulb prosperity, oval or square circular, and middle esophagus ball clack is flourishing, and rear esophageal gland leaf is flourishing, the back side of covering intestines; 4. porus excretorius is arranged in level place after oesophageal bulb; 5. female worm tail is conical, and tail end form is various: blunt circle or carefully round, finger-like or bifurcated or have an abdomen prominent, have or anury point prominent, the prominent form of tail point is various; 6. male worm tail be conical, to outside of belly hook; Copulatory spicules are rose thorn shape, and Ji Ding and base beak are conventionally flourishing, without copulatory bursa.
3, slide the surface sterilization that sword belongs to food fungi nematode: nematode being proceeded in 50 ml centrifuge tubes of sterilizing, is 1.0 g L by quality-volumetric concentration
-1streptomycin sulphate and quality-volumetric concentration be 0.02 g L
-1cycloheximide mixed liquor nematode is carried out to surface sterilization 20 min, 5000 r min
-1centrifugal 5 min, remove supernatant, more repeatedly clean 6 times with sterile water.During each cleaning, fully shake 5000 r min
-1centrifugal 5 min, abandoning supernatant.
4, slide the enrichment culture that sword belongs to food fungi nematode: food fungi nematode is mainly made a living to take food fungi in soil, thereby in pure culture process, needs to provide suitable food source to it.Fungal species is incorrect, and sliding sword genus food fungi nematode is reluctant to take food, nematode mortality; Type of culture medium is incorrect with formula, and fungus breeding is not good enough causes sliding sword genus food fungi nematode not breed by normal growth; The condition such as cultivation temperature, illumination is incorrect, fungi color jaundice, poor growth, easily old and feeble, and nematode is difficult for taking food; The opportunity that nematode is inoculated on medium is incorrect, affects quantity and the quality of nematode food source; The kind of inventor to fungi, the type of medium and formula and condition of culture etc. have carried out a series of groping.The method of determining is as follows.
Adopt conventional method to prepare potato dextrose agar, its culture medium prescription is: in every 1000 ml potato dextrose agars, and potato 220 g, glucose 18 g, agar 18 g, all the other are water, pH value nature.After medium prepares, 121 ℃ of sterilizing 30 min in high-pressure sterilizing pot.After medium is cooling, be poured into the culture dish that diameter is 9 cm, thickness is 0.7-1.0 cm.By fungi Botrytis cinerea (
botrytis cinereapers.) be inoculated on potato dextrose agar, adopt sealed membrane (Parafilm) by culture dish sealing, to prevent the evaporation of medium moisture and to pollute, then culture dish is inverted and is put in constant incubator, 20-25 ℃ of dark culturing, after one week, fungi amount reproduction.Now, sliding sword is belonged to nematode access and cover with in the culture dish of uniformity Botrytis cinerea, inoculation method is in media surface, to include 200 left and right through the food fungi nematode of surface sterilization with suction pipe inoculation 50-100 μ l sterile water in sterile water.Culture dish is adopted to sealed membrane (Parafilm) sealing, to prevent the evaporation of medium moisture and to pollute.Above process is all carried out on aseptic operating platform.Finally, culture dish is faced up and puts into the sterilizing box of suitable size, be then placed in incubator, 20-25 ℃ of dark culturing, after three weeks, Botrytis cinerea disappears substantially, nematode amount reproduction.The sliding sword of each culture dish belongs to food fungi nematode population and is about 12000, and reproduction rate is about 60 times, and individuality is about 0.75 mm, and body length/maximum body breadth is about 35.6, and it is 11.2 μ m that lancet is about.
Choose the medium of the wired worm of a little block length, put into the potato dextrose agar that covers with uniformity Botrytis cinerea, cunning sword is belonged to food fungi nematode and expand numerously, the sliding sword that so repeats to obtain a large amount of pure cultures belongs to eats fungi nematode.
In embodiment, when sliding sword belongs to the enrichment culture of food fungi nematode, the potato dextrose agar formula also adopting comprises: in every 1000 ml potato dextrose agars, potato 190 g, glucose 19 g, agar 19 g, all the other are water, pH value nature.By above-mentioned formula, also obtain sliding sword and belong to food fungi nematode; Under this formula, each culture dish nematode population is about 11000, and reproduction rate is about 55 times, and individuality is about 0.68 mm, and body length/maximum body breadth is about 36.2, and it is 11.0 μ m that lancet is about.
Also adopt in addition following formula: in every 1000 ml potato dextrose agars, potato 200 g, glucose 20 g, agar 20 g, all the other are water, pH value nature.By above-mentioned formula, also obtain sliding sword and belong to food fungi nematode, under this formula, each culture dish nematode population is about 12500, and reproduction rate is about 62.5 times, and individuality is about 0.70 mm, body length/maximum body breadth approximately 35.2, and lancet is about 10.8 μ m.
The sliding sword that has all obtained a large amount of pure cultures under above-mentioned formula condition belongs to food fungi nematode.
5, slide the succeeding preservation that sword belongs to food fungi nematode: the reproduction speed of nematode is very fast, within every 2 weeks, produce a generation, in this process, consume a large amount of hypha,hyphaes and spore, the nutrient of medium and moisture also have certain time limit in addition, thereby the holding time of nematode is also a difficult point.Store method involved in the present invention is as follows.Adopt sealed membrane (Parafilm) by culture dish sealing, then culture dish being faced up, it is standby to be kept in 4 ℃ of refrigerators.Every 5-6 month, sliding sword is belonged to food fungi nematode and carry out subculture cultivation, specific operation process is, culture dish is taken out from 4 ℃ of refrigerators, choose the medium of the wired worm of a little block length, put on the potato dextrose agar that covers with uniformity Botrytis cinerea, to after culture dish sealing, face up and put into sizeable sterilizing box, 25 ℃ of dark culturing, and whether routine observation culture dish be subject to the pollution of other fungies or acarid, after three weeks by culture dish as for preserving in 4 ℃ of refrigerators.
6, slide the application that sword belongs to food fungi nematode: culture dish is taken out from 4 ℃ of refrigerators, with the ox horn spoon of sterilization, medium in culture dish is scraped, put into screen tray, carry out successively the separation of nematode and the sliding sword genus food fungi nematode that surface sterilization step obtains a large amount of pure cultures.
Cunning sword genus food fungi nematode and arbuscular mycorrhizal fungi are inoculated into As polluted soil simultaneously, and the sliding sword of research belongs to interaction and the impact on tobacco growing of eating fungi nematode and arbuscular mycorrhizal fungi.
Result shows, after two inoculations, sliding sword belongs to food fungi nematode population has increased by 1110%, and the mycelia amount of arbuscular mycorrhizal fungi has improved 25%, and Mycorrhizal Infection Incidence also has the raising of some; With do not inoculate sliding sword and belong to food fungi nematode and contrast and compare with the processing of arbuscular mycorrhizal fungi, the interaction that sliding sword belongs to food fungi nematode and arbuscular mycorrhizal fungi significantly improves the biomass (under ground portion dry weight has improved 40%, and acrial part dry weight has improved 66%) of tobacco in As polluted soil and the arsenic cumulant (acrial part arsenic cumulant has improved 16%) in body.Illustrate that sliding sword belongs to food fungi nematode and has superiority in As contaminated soil, promote the growth of plant in contaminated soil, while being applied to aspect the reparation of contaminated soil, can improve phytoremediation efficiency.
Claims (6)
1. the sliding sword of enrichment culture belongs to a method for food fungi nematode, it is characterized in that, concrete steps are as follows:
(1) slide the separation that sword belongs to food fungi nematode: from the fresh soil of field acquisition, adopt improved tray method separated nematode from soil;
(2) slide the evaluation that sword belongs to food fungi nematode: according to sliding sword, belong to the feature of food fungi nematode, under anatomical lens, with capillary, choose this genus nematode, then it is carried out to identification of living body under light microscope;
(3) slide the surface sterilization that sword belongs to food fungi nematode: nematode is proceeded in the centrifuge tube of sterilizing, with streptomycin sulphate and cycloheximide mixed liquor, nematode is carried out to surface sterilization;
(4) slide the enrichment culture that sword belongs to food fungi nematode: by fungi Botrytis cinerea (
botrytis cinereapers.) be inoculated on the potato dextrose agar of sterilizing, adopt sealed membrane that culture dish is sealed, then culture dish is inverted and is put in constant incubator, 20-25 ℃ of dark culturing; After one week, when fungi amount reproduction, sliding sword and belong to nematode access and cover with in the culture dish of uniformity Botrytis cinerea; Culture dish, with after sealed membrane sealing, is placed in sterilizing box, 20-25 ℃ of dark culturing, and routine observation, until grey spore disappears substantially, nematode amount reproduction; Repeat above step and obtain a large amount of sliding sword genus food fungi nematodes; Wherein, the formula of potato dextrose agar is: in every 1000 ml potato dextrose agars, and potato 190-220 g, glucose 18-20 g, agar 15-20 g, all the other are water, pH value nature.
2. method according to claim 1, it is characterized in that: described in step (1), improved tray method is specific as follows: 10 object stainless steel screen traies are put into supporting tray, on screen tray, place two-layer face tissue, then soil sample uniform spreading on paper, in tray, add water until submergence soil; Be placed in again under 20-25 ℃ of temperature, dark condition separated; After 48-60 h, with 400 mesh sieves, collect nematode in beaker; After standing over night, with vavuum pump, slowly draw supernatant liquid, and avoid disturbance lower floor liquid as far as possible, now, lower floor's liquid is collected nematode.
3. method according to claim 1, is characterized in that: in step (3), in described mixed liquor, the quality-volumetric concentration of streptomycin sulphate is 0.80-1.20 g.L
-1; Quality-the volumetric concentration of cycloheximide is 0.015-0.02 g.L
-1; The time of nematode being carried out to surface sterilization is 15-20 min.
4. method according to claim 1, it is characterized in that: in step (2), identify that sliding sword belongs to food fungi nematode, the identification mark of its foundation is as follows: 1. heat kill after death, female worm polypide is until abdomen is curved, the whole polypide of the common afterbody ventrad of male worm hook is " J " shape, and live body nematode is serpentine more; 2. the head contracting of conventionally slightly hanging, headstock osteomalacia, lancet is thinner, and base portion expands, and length is generally 10-12 μ m, is generally no more than 20 μ m; 3. esophagus presoma is cylindrical, middle oesophageal bulb prosperity, oval or square circular, and middle esophagus ball clack is flourishing, and rear esophageal gland leaf is flourishing, the back side of covering intestines; 4. porus excretorius is arranged in level place after oesophageal bulb; 5. female worm tail is conical, and tail end form is various: blunt circle or carefully round, finger-like or bifurcated or have an abdomen prominent, have or anury point prominent, the prominent form of tail point is various; 6. male worm tail be conical, to outside of belly hook; Copulatory spicules are rose thorn shape, and Ji Ding and base beak are conventionally flourishing, without copulatory bursa.
5. method according to claim 1, is characterized in that: the potato dextrose agar through sterilizing described in step (4) be by medium in high-pressure sterilizing pot, at 121 ℃ of temperature, sterilizing 30 min obtain.
6. method according to claim 1, is characterized in that: the store method of the sliding sword genus food fungi nematode obtaining after step (4) enrichment culture is as follows: be placed in culture dish, face up to be placed in 4 ℃ of refrigerators and preserve after sealing; Every 5-6 month, will slide sword and belong to the cultivation of food fungi nematode subculture; Specific operation process is, culture dish is taken out from 4 ℃ of refrigerators, choose the medium of the wired worm of a little block length, put on the potato dextrose agar that covers with uniformity Botrytis cinerea, to after culture dish sealing, put into sterilizing box, 20-25 ℃ of dark culturing, and routine observation, after three weeks by culture dish as for preserving in 4 ℃ of refrigerators.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310696738.1A CN103688908A (en) | 2013-12-18 | 2013-12-18 | Enrichment culture method for aphelenchoidessp. |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310696738.1A CN103688908A (en) | 2013-12-18 | 2013-12-18 | Enrichment culture method for aphelenchoidessp. |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN103688908A true CN103688908A (en) | 2014-04-02 |
Family
ID=50350701
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201310696738.1A Pending CN103688908A (en) | 2013-12-18 | 2013-12-18 | Enrichment culture method for aphelenchoidessp. |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN103688908A (en) |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105766813A (en) * | 2016-04-13 | 2016-07-20 | 江苏省中国科学院植物研究所 | Method for gathering and cultivating aphelenchoidessp |
| CN105821112A (en) * | 2016-04-27 | 2016-08-03 | 江苏省中国科学院植物研究所 | Method for studying feeding preference of soil fungal-feeding nematodes |
| CN109799326A (en) * | 2019-03-03 | 2019-05-24 | 中国科学院亚热带农业生态研究所 | A method of estimating different Feeding Habits Soil nematode biomass |
| 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 (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101473808A (en) * | 2009-01-22 | 2009-07-08 | 中国热带农业科学院热带生物技术研究所 | Artificial culture, breeding and preservation method of plant root-knot nematode |
| CN101831388A (en) * | 2010-05-18 | 2010-09-15 | 华南农业大学 | Nematophagous fungus and preparation method and application thereof |
| CN102972355A (en) * | 2012-12-05 | 2013-03-20 | 华南农业大学 | Method for breeding, culturing and storing plant nematodes by using sweet potato calluses |
-
2013
- 2013-12-18 CN CN201310696738.1A patent/CN103688908A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101473808A (en) * | 2009-01-22 | 2009-07-08 | 中国热带农业科学院热带生物技术研究所 | Artificial culture, breeding and preservation method of plant root-knot nematode |
| CN101831388A (en) * | 2010-05-18 | 2010-09-15 | 华南农业大学 | Nematophagous fungus and preparation method and application thereof |
| CN102972355A (en) * | 2012-12-05 | 2013-03-20 | 华南农业大学 | Method for breeding, culturing and storing plant nematodes by using sweet potato calluses |
Non-Patent Citations (3)
| Title |
|---|
| 段玉玺: "《植物线虫学》", 31 July 2011, 中国农业出版社 * |
| 毛小芳等: "土壤线虫三种分离方法效率比较", 《生态学杂志》 * |
| 毛小芳等: "土壤食细菌线虫的原位富集培养方法", 《生态学报》 * |
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105766813A (en) * | 2016-04-13 | 2016-07-20 | 江苏省中国科学院植物研究所 | Method for gathering and cultivating aphelenchoidessp |
| CN105766813B (en) * | 2016-04-13 | 2018-10-09 | 江苏省中国科学院植物研究所 | A kind of method that enrichment culture slides sword category Fungal-feeding nematode |
| CN105821112A (en) * | 2016-04-27 | 2016-08-03 | 江苏省中国科学院植物研究所 | Method for studying feeding preference of soil fungal-feeding nematodes |
| CN109799326A (en) * | 2019-03-03 | 2019-05-24 | 中国科学院亚热带农业生态研究所 | A method of estimating different Feeding Habits Soil nematode biomass |
| CN109799326B (en) * | 2019-03-03 | 2021-12-14 | 中国科学院亚热带农业生态研究所 | Method for estimating biomass of nematodes in different feeding soil |
| 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 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN106834087B (en) | A kind of the AM fungi numerous and functional experiment device of expansion and application method of timing cycle | |
| CN101411321B (en) | Method for artificially feeding Bemisia tabaci Gennadius | |
| Gibor | Some ecological relationships between phyto-and zooplankton | |
| Adamec | Functional characteristics of traps of aquatic carnivorous Utricularia species | |
| CN103688908A (en) | Enrichment culture method for aphelenchoidessp. | |
| CN110200016B (en) | Streptomyces strain PBSH9 capable of preventing and treating potato scab and promoting potato growth and application thereof | |
| CN106212281B (en) | A kind of method for tissue culture for improving banana survival rate | |
| CN104026078B (en) | A kind of reticulitermes flavipe indoor intensive raising method | |
| Couch | The formation and operation of the traps in the nematode-catching fungus, Dactylella bembicodes Drechsler | |
| CN105918168A (en) | Exopalaemon carinicauda saline-alkaline water artificial culturing method | |
| CN106359206B (en) | Seed conservation culture method for daphnia magna | |
| CN107996523A (en) | The method of indoor rapid amplifying breeding brassicaceous vegetable lepidoptera pest parasitic wasp | |
| CN109258304A (en) | A kind of production method and growth condition of hickory chick liquid culture bacteria | |
| CN106635821A (en) | Mycorrhiza fungi for promoting nutrient absorption of blueberry and preparation method and application of mycorrhiza fungi | |
| CN106222121A (en) | A kind of bacillus megaterium bacterial strain, biocontrol agent and preparation method and application | |
| CN102732430A (en) | Aspergillus niger strain and application thereof | |
| CN112753658A (en) | Method for indoor breeding trichogramma dendrolimi by utilizing bombyx camphorata eggs | |
| CN107810926A (en) | The artificial large-scale breeding method of Wan Shi Eimeria aythyaes | |
| CN108913625B (en) | Salt-tolerant streptomycete, microbial inoculum thereof and application of microbial inoculum thereof in promoting plant growth | |
| CN103756913A (en) | Isaria fumosorosea | |
| CN1325630C (en) | Culture of nematoda without bacteria | |
| CN105532583A (en) | Living body breeding method for entomopathogenic nematodes | |
| CN106069159B (en) | The inoculation method of blood aspidistra | |
| CN109496706A (en) | A kind of sterile brachyplast of Mao Xu Cao quickly breed before explant preprocess method | |
| CN105766813B (en) | A kind of method that enrichment culture slides sword category Fungal-feeding nematode |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C02 | Deemed withdrawal of patent application after publication (patent law 2001) | ||
| WD01 | Invention patent application deemed withdrawn after publication |
Application publication date: 20140402 |