CN103529194A - Method for detecting drug resistance of green peach aphid - Google Patents
Method for detecting drug resistance of green peach aphid Download PDFInfo
- Publication number
- CN103529194A CN103529194A CN201310477050.4A CN201310477050A CN103529194A CN 103529194 A CN103529194 A CN 103529194A CN 201310477050 A CN201310477050 A CN 201310477050A CN 103529194 A CN103529194 A CN 103529194A
- Authority
- CN
- China
- Prior art keywords
- medicament
- peach aphid
- glass tube
- population
- diagnostic dose
- 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
Images
Landscapes
- Investigating Or Analysing Biological Materials (AREA)
Abstract
The invention provides a method for detecting the drug resistance of green peach aphid. Diagnosis glass tubes are produced under the diagnosis dosage of medicaments, and the resistance level of field green peach aphid population to the medicaments is rapidly detected, wherein the medicaments refer to pirimor, bifenthrin, imidacloprid or acetamiprid, the diagnosis dosage of pirimor is 10086.30ng/cm2, bifenthrin is 2159.00ng/cm2, imidacloprid is 47.86ng/cm2 and acetamiprid is 196.58ng/cm2. The method is simple and convenient in use, intuitive and clear in results, suitable for rapidly detecting the resistance level of field green peach aphid, overcomes the defects of large sample consumption amount, complex operation and time consumption of the traditional detection method, and can be used for rapidly detecting the resistance level of field green peach aphid population.
Description
Technical field
The present invention relates to a kind of determination techniques method that the agricultural pests resistance to the action of a drug is monitored, specifically, is to detect black peach aphid to the drug-fast method of four kinds of insecticides (Aphox, Biphenthrin, Imidacloprid and Acetamiprid) with diagnostic dose of the present invention.
Background technology
Black peach aphid is one of the widest of paramount importance insect that distributes in the world, can in 50Duo Ge section 400 various plants, take food, not only can directly sting suction plant nutrient, cause the serious dehydration of plant and malnutrition, all right transmitted virus, plant is caused to larger consequential injury, agricultural production is caused to huge economic loss.For a long time, the control of black peach aphid mainly be take to chemical prevention as main, a large amount of, the lasting and unreasonable use due to chemical insecticide, causes the black peach aphid of many countries and regions to produce the resistance to the action of a drug in various degree to common insecticide.
The drug-fast formation and development of black peach aphid has caused very large difficulty to chemical prevention, and it is very important how detecting fast and accurately black peach aphid Detection of insecticide resistance.Method for monitoring for resistance mainly comprises traditional biologicall test, biochemistry detection method, immunological method and molecular biology method now.Generally, biologicall test is the most directly, approaches the method for Fields detection most.
Difference due to various pharmacy effect mechanism, in short time, the toxic reaction of insect is also different, therefore utilize the glass tube medicine film diagnostic dose of this feature development, resistance level that can fast detecting field population, this application is also the important means of field fast detecting resistance.
Toxicity test is the conventional method of indoor understanding pest resistance to insecticide, but traditional immersion method and drop method carry out toxicity test, and complex operation, set some concentration and repetition, to guarantee the confidence level of result.Glass tube medicine embrane method is the method for emerging a kind of fast detecting pest resistance to insecticide, because its action effect is obvious, has shortened the time of check result, but still the same with classic method with in replication problem for concentration, does not improve.Utilize diagnostic dose of the present invention, be aided with glass tube medicine embrane method, not only utilized the fast detecting feature of glass tube medicine embrane method, and can understand fast pest resistance under diagnostic dose, the traditional toxicity test experiment red tape solving, improve efficiency, and can be used for the drug-fast fast detecting of Field Pests.
Summary of the invention
The object of this invention is to provide a kind of drug-fast method of detection black peach aphid, make diagnosis glass tube under the diagnostic dose of medicament, detect the resistance level of black peach aphid field population to medicament, described medicament is Aphox, Biphenthrin, Imidacloprid and Acetamiprid; The diagnostic dose of medicament: Aphox is 10086.30ng/cm
2, Biphenthrin is 2159.00ng/cm
2, Imidacloprid is 47.86ng/cm
2, Acetamiprid is 196.58ng/cm
2.
Mixed population is that various places gather black peach aphid population, after mixing, raise many generations, therefore the black peach aphid that contains each resistance level in this population is individual, on raw survey, enzyme work and molecular level, different resistances have all been showed, using this population as experimental population of the present invention, more approach field population, experimental result more reliably, more accurate.
Wherein, the diagnostic dose of medicament is that the medicament by variable concentrations adopts glass tube medicine embrane method to carry out Study on Fungicide Toxicity to indoor feeding mixed population, calculates LC
99diagnostic dose as medicament.
Above-mentioned indoor feeding mixed population is for gathering black peach aphid field, various places population, and after mixing, indoor feeding is many for obtaining.
Further, the diagnostic dose of medicament is determined as follows:
(1) indoor feeding of black peach aphid field population: gather black peach aphid field, various places population, mix and raise many generations, obtain indoor feeding mixed population;
(2) four kinds of insecticides of acetone diluted are that different concentration gradient is as toxicity test concentration;
(3) making of glass tube medicine film: with each gradient pharmaceutical production toxicity test glass tube medicine film used of acetone diluted in step (2), every pipe adds 220 each concentration liquids of μ L, rolling glass tube to liquid is evenly adsorbed on glass tube inwall, until acetone volatilizees completely;
(4) toxicity test: carry out the toxicity test of black peach aphid with the glass tube of preparation in step (3), at least carry out three secondary pollutants and learn repetition;
(5) result and diagnostic dose: the result to step (4) is done toxicity regression curve, and calculates LC
99draw the diagnostic dose of each medicament.
Wherein, in step (2), insecticide used is more than 95% former medicine, and with acetone solution, is diluted to 5～7 concentration gradients.
Wherein, in step (4), simple independent repetition done in each toxicity test experiment three times.
Wherein, POLO software result of calculation and diagnostic dose for step (5).
Beneficial effect of the present invention is:
The inventive method is easy to use, and visual result understands, is applicable to field fast detecting black peach aphid resistance level, has overcome that traditional raw survey method is large with sample size, complex operation, the shortcoming such as time-consuming, can be used for fast detecting black peach aphid field population resistance level.
Accompanying drawing explanation
Fig. 1 is experiment glass tube used.
The aptery one-tenth aphid that Fig. 2 selects for experiment.
Fig. 3 is for producing the black peach aphid of toxicity symptom.
Embodiment
Following examples are used for illustrating the present invention, but are not used for limiting the scope of the invention.
The acquisition of embodiment 1 diagnostic dose
1. research object
Research object of the present invention is the laboratory rearing various places black peach aphid mixed population in many generations.
2. method
Reagent agent Aphox, Biphenthrin, Imidacloprid and Acetamiprid, use glass tube medicine embrane method, carries out toxicity test.
The configuration of 2.1 former medicine mother liquors and dilution
Former medicine is dissolved in acetone, is loaded in flask for medicinal preparations after constant volume, sealed membrane sealing, is stored in 4 ℃ of refrigerators.
Before toxicity test, with acetone, former medicine mother liquor is diluted to each concentration, standby.
The making of 2.2 glass tube medicine films
Testing glass tube used (as shown in Figure 1) must, through strict washing, guarantee without any medicament residue.From the liquid having diluted, draw 220 μ L and join diameter 1.7cm, glass tube (the internal surface area 40cm of high 7.5cm
2) in, rolling glass tube to liquid is evenly adsorbed on glass tube inwall, until acetone volatilizees completely, acetone treatment, the use of spending the night under the glass tube medicine film room temperature of making are used separately in contrast.
2.3 toxicity tests are for the selection of examination aphid
Select the black peach aphid population of indoor feeding, the healthy consistent aptery one-tenth aphid (as shown in Figure 2) of picking is tested, and the medicine film pipe that each prepares is placed 20 aphids, (20～25 ℃ of temperature under indoor normal raising condition, relative humidity 50%～75%, illumination 16L:8D).
The judgement of 2.4 check results and death standard
Check result after 3h, wherein only have one leg moving or completely motionless be designated as dead (as shown in Figure 3), take and contrast mortality ratio and be less than 10% for effective mensuration, and proofread and correct by contrast mortality ratio.6～7 concentration of each medicament, each concentration repeats for 3 times, and data processing adopts POLO analysis software, calculates LC
99, i.e. diagnostic dose, this diagnostic dose can be used for the resistance level of fast detecting black peach aphid field population to Aphox, Biphenthrin, Imidacloprid and Acetamiprid.
The diagnostic dose of medicament is Aphox 10086.3ng/cm
2, Biphenthrin 2159.0ng/cm
2, Imidacloprid 47.9ng/cm
2, Acetamiprid 196.6ng/cm
2.
Embodiment 2 diagnostic doses detect the black peach aphid resistance to the action of a drug
1. research object
This case study object is field population to be measured.
2. method
Reagent agent Aphox, Biphenthrin, Imidacloprid and Acetamiprid, utilize diagnostic dose of the present invention to make glass tube medicine film, and the detection of black peach aphid resistance level is carried out in field.
The configuration of 2.1 former medicine mother liquors and dilution
Former medicine is dissolved in acetone, the diagnostic dose former medicine mother liquor dilution example 1 being obtained with acetone, standby.
The making of 2.2 glass tube medicine films
Testing glass tube used (as shown in Figure 1) must, through strict washing, guarantee without any medicament residue.From the liquid having diluted, draw 220 μ L and join diameter 1.7cm, glass tube (the internal surface area 40cm of high 7.5cm
2) in, rolling glass tube to liquid is evenly adsorbed on glass tube inwall, until acetone volatilizees completely, and, contrast is separately by acetone treatment, the use of spending the night under the glass tube medicine film room temperature of making.
2.3 field resistance diagnosis
Field is selected aptery one-tenth aphid of the same size and is placed in 2.2 making glass tubes, and 20 of every pipes, observe mortality ratio after 3h, and mortality ratio is resistant population lower than 99% population.
Utilize diagnostic dose of the present invention to carry out field resistance diagnosis, not only greatly reduced the loaded down with trivial details step of traditional toxicity test, and also convenient and reliable for the fast detecting in field, solved the large problem of worm amount for traditional toxicity test method, also solved field operational issue, simple to operation.For detecting in real time resistance in field, understand resistance level, and then instruct field medication.
Embodiment 3 traditional glass pipe medicine embrane methods carry out drug-fast mensuration to field black peach aphid.
1. research object
This case study object is field population to be measured.
2. method
Reagent agent Aphox, Biphenthrin, Imidacloprid and Acetamiprid, use glass tube medicine embrane method, carries out toxicity test.
The configuration of 2.1 former medicine mother liquors and dilution
Former medicine is dissolved in acetone, is loaded in flask for medicinal preparations after constant volume, sealed membrane sealing, is stored in 4 ℃ of refrigerators.
Before toxicity test, with acetone, former medicine mother liquor is diluted to each concentration, standby.
The making of 2.2 glass tube medicine films
Testing glass tube used (as shown in Figure 1) must, through strict washing, guarantee without any medicament residue.From the liquid having diluted, draw 220 μ L and join diameter 1.7cm, glass tube (the internal surface area 40cm of high 7.5cm
2) in, rolling glass tube to liquid is evenly adsorbed on glass tube inwall, until acetone volatilizees completely, acetone treatment, the use of spending the night under the glass tube medicine film room temperature of making are used separately in contrast.
2.3 toxicity tests are for the selection of examination aphid
Select the black peach aphid population of indoor feeding, the healthy consistent aptery one-tenth aphid (as shown in Figure 2) of picking is tested, and the medicine film pipe that each prepares is placed 20 aphids, (20～25 ℃ of temperature under indoor normal raising condition, relative humidity 50%～75%, illumination 16L:8D).
The judgement of 2.4 check results and death standard
Check result after 3h, wherein only have one leg moving or completely motionless be designated as dead (as shown in Figure 3), take and contrast mortality ratio and be less than 10% for effective mensuration, and proofread and correct by contrast mortality ratio.6～7 concentration of each medicament, each concentration repeats for 3 times, and data processing adopts POLO analysis software.
The raw survey method of traditional glass pipe medicine film, complex operation step, the multiple operation steps of every secondary check weighing, and large by worm amount, be not suitable for field and detect in real time.Adopt diagnostic dose of the present invention, make glass tube, can take field to and detect in real time, understand fast field population resistance.
Adopt traditional glass pipe medicine embrane method to gather black peach aphid population to field and carry out toxicity test, tested population is: Dalian, Langfang in Hebei Province, Minxing, Shanghai.Classic method is measured and is obtained the LC of three tested populations to four kinds of medicaments
50, see following form.Adopt glass tube diagnostic dose of the present invention three tested populations to be carried out to the resistance diagnosis of above four kinds of medicaments, method of operating is as embodiment 2 simultaneously, and the mortality ratio of each population to diagnostic medicament, the results are shown in Table 1.
By list data, can be found out, the resistance level that the mortality ratio that glass tube medicine film diagnostic dose calculates and traditional toxicity test method obtain has obvious correlativity, represent also can detect field population resistance level with the diagnostic dose of this patent, but the method has overcome, and traditional raw survey method sample size is large, complex operation, the shortcoming such as time-consuming, is more suitable for Fields detection.
The resistance diagnosis of four kinds of medicament glass tube diagnostic doses of table 1
Although above the present invention is described in detail with a general description of the specific embodiments, on basis of the present invention, can make some modifications or improvements it, this will be apparent to those skilled in the art.Therefore, these modifications or improvements, all belong to the scope of protection of present invention without departing from theon the basis of the spirit of the present invention.
Claims (7)
1. detect the drug-fast method of black peach aphid, make diagnosis glass tube under the diagnostic dose of medicament, detect the resistance level of black peach aphid field population at individual to medicament, it is characterized in that, described medicament is Aphox, Biphenthrin, Imidacloprid or Acetamiprid; The diagnostic dose of medicament: Aphox is 10086.30ng/cm
2, Biphenthrin is 2159.00ng/cm
2, Imidacloprid is 47.86ng/cm
2, Acetamiprid is 196.58ng/cm
2.
2. method according to claim 1, is characterized in that, the diagnostic dose of medicament is that the medicament by variable concentrations adopts glass tube medicine embrane method to carry out Study on Fungicide Toxicity to indoor feeding mixed population, calculates LC
99diagnostic dose as medicament.
3. method according to claim 2, is characterized in that, described indoor feeding mixed population is for gathering black peach aphid field, various places population, and after mixing, indoor feeding is many for obtaining.
4. according to the method described in claim 1-3 any one, it is characterized in that, the diagnostic dose of medicament is determined as follows:
(1) indoor feeding of black peach aphid field population: gather black peach aphid field, various places population, mix and raise many generations, obtain indoor feeding mixed population;
(2) four kinds of insecticides of acetone diluted to variable concentrations gradient as toxicity test concentration;
(3) making of glass tube medicine film: with each gradient pharmaceutical production toxicity test glass tube medicine film used of acetone diluted in step (2), every pipe adds 220 each concentration liquids of μ L, rolling glass tube, until acetone volatilizees completely, is evenly distributed on glass tube inwall liquid;
(4) toxicity test: carry out the toxicity test of black peach aphid with the glass tube of preparation in step (3), at least carry out three secondary pollutants and learn repetition;
(5) result and diagnostic dose: the result to step (4) is done toxicity regression curve, and calculates LC
99draw the diagnostic dose of each medicament.
5. method according to claim 4, is characterized in that, in step (2), insecticide used is more than 95% former medicine, and with acetone solution, is diluted to 5～7 concentration gradients.
6. method according to claim 4, is characterized in that, in step (4), simple independent repetition done in each toxicity test experiment three times.
7. method according to claim 4, is characterized in that, POLO software result of calculation and diagnostic dose for step (5).
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310477050.4A CN103529194A (en) | 2013-10-12 | 2013-10-12 | Method for detecting drug resistance of green peach aphid |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310477050.4A CN103529194A (en) | 2013-10-12 | 2013-10-12 | Method for detecting drug resistance of green peach aphid |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN103529194A true CN103529194A (en) | 2014-01-22 |
Family
ID=49931373
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201310477050.4A Pending CN103529194A (en) | 2013-10-12 | 2013-10-12 | Method for detecting drug resistance of green peach aphid |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN103529194A (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103776992A (en) * | 2014-01-26 | 2014-05-07 | 青岛农业大学 | Monitoring method for imidacloprid drug resistance of apple aphis citricola |
| CN109287394A (en) * | 2018-11-27 | 2019-02-01 | 云南省农业科学院花卉研究所 | Between plant Dalmatian chrysanthemum prevention and control rose insect pest method |
-
2013
- 2013-10-12 CN CN201310477050.4A patent/CN103529194A/en active Pending
Non-Patent Citations (4)
| Title |
|---|
| GRAHAM D.MOORES ET AL: "Characterisation of Insensitive Acetylcholinesterase in Insecticide-Resistant Cotton Aphids, Aphis gossypii Glover (Homoptera: Aphididae)", 《PESTICIDE BIOCHEMISTRY AND PHYSIOLOGY》 * |
| 宫亚军等: "北京地区不同桃蚜种群的抗药性研究", 《中国农业科学》 * |
| 高希武等: "北京地区桃蚜抗抗药性研究初报", 《农药》 * |
| 鲁艳辉等，: "禾谷缢管蚜和麦长管蚜玻璃管药膜法敏感毒力基线的建立", 《昆虫学报》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103776992A (en) * | 2014-01-26 | 2014-05-07 | 青岛农业大学 | Monitoring method for imidacloprid drug resistance of apple aphis citricola |
| CN109287394A (en) * | 2018-11-27 | 2019-02-01 | 云南省农业科学院花卉研究所 | Between plant Dalmatian chrysanthemum prevention and control rose insect pest method |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN202166640U (en) | Kit for quickly detecting virulence of pesticide to Bemisia tabaci and emulsion glass tube | |
| CN104155441B (en) | A kind of pesticide residue detection method based on micro-fluidic chip | |
| CN103675208A (en) | Method for detecting drug resistance of adelphocoris suturalis | |
| CN103529194A (en) | Method for detecting drug resistance of green peach aphid | |
| CN103698498A (en) | Method for measuring pesticide resistance of macrosiphum avenae | |
| CN104407125B (en) | A kind of method and test kit being suitable for the responsive cotten aphid detection and identification of land for growing field crops cotton planting | |
| CN103675254B (en) | Method for detecting drug resistance of rhopalosiphum aphids | |
| CN104770340B (en) | A kind of method that utilization easy device determines citrus fruit fly stomach toxicity virulence | |
| CN103630633A (en) | Method for measuring contents of six amino acids in ixeris sonchifolia injection by using pre-column derivatization method | |
| CN106872618B (en) | Pyrethroid pesticide remained extracting process in liquid sample | |
| CN202126438U (en) | Automatic analyzer for platelets | |
| CN103675253A (en) | Method for detecting insecticide resistance of apolygus lucorum | |
| CN102050876A (en) | Positive-serum and negative-serum standard substance of avian influenza virus H5N1 subtype Re-5 strain and preparation method thereof | |
| CN107167583B (en) | Method for detecting drug resistance of chilo suppressalis | |
| CN103115985B (en) | Method for synchronously detecting six flavonoid constituents in sowthistle-leaf ixeris seedling injection in HPLC-DAD (High Performance Liquid Chromatography-Diode Array detector) method | |
| CN206573589U (en) | Crop fungal spore liquefaction resistance device | |
| CN101975855A (en) | Reagent and method for detecting efficacy on inactivated vaccine against duck infectious serositis | |
| CN103543234B (en) | High-performance liquid chromatography analysis method for simultaneously detecting contents of chlorpyrifos and beta-cypermethrin | |
| CN109459538A (en) | The quickly kit and its method of screening prevention and treatment brown paddy plant hopper insecticide | |
| CN105137054B (en) | A kind of method of wheat aphid indoor bioassay | |
| CN111362898A (en) | Method for optimizing extraction of perfoliote knotweed herb quercetin by Plackett-Burnman combined response surface method | |
| CN107015014A (en) | Crop fungal spore liquefaction resistance device and detection method | |
| CN103385867B (en) | Application of sesquiterpenoids in preparing mite killing medicine | |
| CN103315008B (en) | Preparation method of Suaeda salsa used as agricultural pesticide and application | |
| CN104673877B (en) | One mycoplasma species Chinese medicine minimum inhibitory concentration new detecting method |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C12 | Rejection of a patent application after its publication | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20140122 |