CN103698498A - Method for measuring pesticide resistance of macrosiphum avenae - Google Patents
Method for measuring pesticide resistance of macrosiphum avenae Download PDFInfo
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- CN103698498A CN103698498A CN201310682351.0A CN201310682351A CN103698498A CN 103698498 A CN103698498 A CN 103698498A CN 201310682351 A CN201310682351 A CN 201310682351A CN 103698498 A CN103698498 A CN 103698498A
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Abstract
The invention provides a method for measuring the pesticide resistance of macrosiphum avenae. The method comprises the following steps of manufacturing a diagnosis glass pipe under the diagnosis dosage of a pesticide, rapidly detecting the resistance level to the pesticide from macrosiphum avenae field population, wherein the pesticide is imidacloprid, pirimor, omethoate or beta-cypermethrin, and the diagnosis dosage of the pesticide comprises 4289ng/cm<2> of the imidacloprid, 103ng/cm<2> of the pirimor, 224ng/cm<2> of the omethoate, and the 4695ng/cm<2> of the beta-cypermethrin. According to the method provided by the invention, the use is simple and convenient, the result is intuitive, the method is suitable for rapidly detecting macrosiphum avenae resistance level in the field, the disadvantages that the number of samples in the traditional detecting method is large, the operation is tedious, the time is wasted and the like are overcome, and the method is used for rapidly detecting the macrosiphum avenae field resistance level.
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, relate to a kind of drug-fast method of detection grain aphid.
Background technology
Grain aphid is one of Important Agricultural insect of the global wheat of causing harm, each wheat producing countries all over the world, and in China, north and south Mai Qu all has generation.Grain aphid not only can directly be stung suction plant nutrient, causes the serious dehydration of plant and malnutrition, and all right transmitted virus, causes larger consequential injury to plant, and agricultural production is caused to huge economic loss.For a long time, the control of grain aphid mainly be take to chemical prevention as main, a large amount of, the lasting and unreasonable use due to chemical insecticide, causes the grain 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 grain aphid has caused very large difficulty to chemical prevention, and it is very important how detecting fast and accurately grain 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 easy to usely, visual result understands, is applicable to the method for field fast detecting grain aphid resistance level.
In order to realize object of the present invention, the invention provides a kind of drug-fast method of detection grain aphid, under the diagnostic dose of medicament, make diagnosis glass tube, detect the resistance level of grain aphid field population to medicament, described medicament is Imidacloprid, Aphox, flolimat or effective cypermethrin; The diagnostic dose of medicament: Imidacloprid is 4289ng/cm
2, Aphox is 103ng/cm
2, flolimat is 224ng/cm
2, effective cypermethrin is 4695ng/cm
2.
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 grain aphid field, various places population, and after mixing, indoor feeding is many for obtaining.Mixed population is that various places gather grain aphid population, after mixing, raise many generations, therefore the grain 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.
Further, the diagnostic dose of medicament is determined as follows:
(1) indoor feeding of grain aphid field population: gather grain 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 grain 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, visual result understands, be applicable to field fast detecting grain aphid resistance level, 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 grain aphid field population resistance level.
Accompanying drawing explanation
Fig. 1 is the present invention's diagnosis glass tube used;
Fig. 2 is that the present invention tests the aptery one-tenth aphid of selecting.
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 grain aphid mixed population in many generations.
2. method
Reagent agent Imidacloprid, Aphox, flolimat and effective cypermethrin, 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 grain 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 deadly, 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, and 4 kinds of insecticides are as shown in table 1 to the toxicity test result of grain aphid, calculate LC
99, i.e. diagnostic dose, this diagnostic dose can be used for the resistance level of fast detecting grain aphid field population to Imidacloprid, Aphox, flolimat and effective cypermethrin.
The toxicity test result of four kinds of insecticides of table 1 to grain aphid
N represents the borer population that toxicity test is used.
The diagnostic dose of medicament: Imidacloprid is 4289ng/cm
2, Aphox is 103ng/cm
2, flolimat is 224ng/cm
2, effective cypermethrin is 4695ng/cm
2.
Embodiment 2 diagnostic doses detect the grain aphid resistance to the action of a drug
1. research object
This case study object is field grain aphid population to be measured.
2. method
Reagent agent Imidacloprid, Aphox, flolimat and effective cypermethrin, utilize diagnostic dose of the present invention to make glass tube medicine film, and the detection of grain 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, 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 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.
Advantage:
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 chemical control diagnosis glass tubes carry out drug-fast mensuration to field grain aphid
1. research object
This case study object is field grain aphid population to be measured.
2. method
Reagent agent Imidacloprid, Aphox, flolimat and effective cypermethrin, utilize diagnostic dose of the present invention to make glass tube medicine film, and field grain aphid is carried out to drug-fast mensuration.
2.1 operation steps
1), by field sampling standard, extract the wheat plant with aphid, and take back indoor;
2) concussion wheat seeding comes off on blank sheet of paper aphid, and choose active grain aphid and put into diagnosis glass tube, every pipe 10-20 head (as far as possible choosing aptery one-tenth aphid), gauze sealing, each medicament 3-5 diagnostic reagent pipe repeats;
3) by being housed, (avoiding direct sunlight) in the diagnostic reagent pipe holding chamber of aphid check mortality ratio after 3 hours.
2.2 test site
Laizhou City, Shandong Province, Wenshang County, Boxing County
2.3 discrimination standard
Mortality ratio reaches following standard (table 2) and illustrates that this medicament can effectively control field population.Lower than following standard, do not mean that there is no prevention effect, can strengthen dispenser dosage (being generally the upper limit of RD).
The dead criterion of table 2
2.4 results and analysis
As can be seen from Table 3, Laizhou City grain aphid mortality ratio in flolimat, Aphox, Imidacloprid and 4 kinds of medicament diagnostic reagent pipes of effective cypermethrin is respectively 94%, 96%, 93%, 100%, all, higher than its discrimination standard, show that testing medicaments for four kinds all can effectively control the grain aphid population of this county; Wenshang County grain aphid mortality ratio in Imidacloprid diagnostic reagent pipe is 86.1% higher than discrimination standard, illustrate that Imidacloprid can effectively control this county grain aphid population, and grain aphid mortality ratio in flolimat, Aphox, effective cypermethrin diagnostic reagent pipe is respectively 86.0%, 93.3%, 86.2%, all lower than discrimination standard, illustrate that this county grain aphid has produced certain resistance to the action of a drug to above three kinds of medicaments, when actual medication, dispenser dosage can be strengthened, but the maximum recommended dosage on label should not be surpassed; The mortality ratio of Boxing County grain aphid in flolimat, Aphox, Imidacloprid diagnosis glass tube is respectively 92.1%, 100%, 100%, all higher than discrimination standard, illustrate that three kinds of medicaments can effectively control this county grain aphid population, and the mortality ratio of this county grain aphid in effective cypermethrin diagnosis glass tube is 55.7%, significantly all, lower than discrimination standard, suggestion this county can be postponed the use of effective cypermethrin.
Table 3 is the mortality ratio in four kinds of medicament diagnosis glass tubes for examination worm
Embodiment 4 traditional glass pipe medicine embrane methods are measured the resistance to the action of a drug of field grain aphid population
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 grain aphid population to field and carry out toxicity test, tested population is: Distributions in Liaocheng of Shandong Province, Yuncheng, Shanxi, the Bei Jingshang village.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 six 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 4.
The resistance diagnosis of four kinds of medicament glass tube diagnostic doses of table 4
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.
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. one kind is detected the drug-fast method of grain aphid, under the diagnostic dose of medicament, make diagnosis glass tube, detect the resistance level of grain aphid field population at individual to medicament, it is characterized in that described medicament Imidacloprid, Aphox, flolimat or effective cypermethrin; The diagnostic dose of medicament: Imidacloprid is 4289ng/cm
2, Aphox is 103ng/cm
2, flolimat is 224ng/cm
2, effective cypermethrin is 4695ng/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 grain 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 grain aphid field population: gather grain 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: the toxicity test of carrying out grain aphid with the glass tube of preparation in step (3);
(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 6～7 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).
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106093368A (en) * | 2016-06-20 | 2016-11-09 | 中国农业科学院植物保护研究所 | Quickly measure melon aphid to the diagnostic kit of Acetamiprid sensitivity and using method |
| CN110514782A (en) * | 2019-05-20 | 2019-11-29 | 中国农业科学院蔬菜花卉研究所 | A kind of bioassay method of fast and convenient aleyrodid class stomach insecticide |
-
2013
- 2013-12-12 CN CN201310682351.0A patent/CN103698498A/en active Pending
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| GRAHAM D. MOORES ET AL: "Characterisation of Insensitive Acetylcholinesterase in Insecticide-Resistant Cotton Aphids, Aphis gossypii Glover (Homoptera: Aphididae)", 《PESTICIDE BIOCHEMISTRY AND PHYSIOLOGY》 * |
| 师桂英等: "春小麦种质对麦长管蚜的抗蚜性鉴定", 《兰州大学学报（自然科学版）》 * |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106093368A (en) * | 2016-06-20 | 2016-11-09 | 中国农业科学院植物保护研究所 | Quickly measure melon aphid to the diagnostic kit of Acetamiprid sensitivity and using method |
| CN110514782A (en) * | 2019-05-20 | 2019-11-29 | 中国农业科学院蔬菜花卉研究所 | A kind of bioassay method of fast and convenient aleyrodid class stomach insecticide |
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Application publication date: 20140402 |