CN107167583A - Method for detecting drug resistance of chilo suppressalis - Google Patents
Method for detecting drug resistance of chilo suppressalis Download PDFInfo
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- CN107167583A CN107167583A CN201710614975.7A CN201710614975A CN107167583A CN 107167583 A CN107167583 A CN 107167583A CN 201710614975 A CN201710614975 A CN 201710614975A CN 107167583 A CN107167583 A CN 107167583A
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- made feeds
- rice borer
- striped rice
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- 238000000034 method Methods 0.000 title claims abstract description 49
- 241000426497 Chilo suppressalis Species 0.000 title claims abstract description 46
- 206010059866 Drug resistance Diseases 0.000 title abstract 3
- 239000003814 drug Substances 0.000 claims abstract description 74
- AMFGTOFWMRQMEM-UHFFFAOYSA-N triazophos Chemical group N1=C(OP(=S)(OCC)OCC)N=CN1C1=CC=CC=C1 AMFGTOFWMRQMEM-UHFFFAOYSA-N 0.000 claims abstract description 17
- PSOVNZZNOMJUBI-UHFFFAOYSA-N chlorantraniliprole Chemical compound CNC(=O)C1=CC(Cl)=CC(C)=C1NC(=O)C1=CC(Br)=NN1C1=NC=CC=C1Cl PSOVNZZNOMJUBI-UHFFFAOYSA-N 0.000 claims abstract description 16
- 231100000419 toxicity Toxicity 0.000 claims abstract description 11
- 230000001988 toxicity Effects 0.000 claims abstract description 11
- 229940079593 drug Drugs 0.000 claims abstract description 8
- 238000001514 detection method Methods 0.000 claims description 16
- CGQCWMIAEPEHNQ-UHFFFAOYSA-N Vanillylmandelic acid Chemical compound COC1=CC(C(O)C(O)=O)=CC=C1O CGQCWMIAEPEHNQ-UHFFFAOYSA-N 0.000 claims description 15
- ARUVKPQLZAKDPS-UHFFFAOYSA-L copper(II) sulfate Chemical compound [Cu+2].[O-][S+2]([O-])([O-])[O-] ARUVKPQLZAKDPS-UHFFFAOYSA-L 0.000 claims description 15
- 229910000366 copper(II) sulfate Inorganic materials 0.000 claims description 15
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 14
- ZMXDDKWLCZADIW-UHFFFAOYSA-N N,N-Dimethylformamide Chemical compound CN(C)C=O ZMXDDKWLCZADIW-UHFFFAOYSA-N 0.000 claims description 12
- 238000003756 stirring Methods 0.000 claims description 12
- 239000000126 substance Substances 0.000 claims description 12
- 238000002360 preparation method Methods 0.000 claims description 11
- 231100000820 toxicity test Toxicity 0.000 claims description 11
- 239000011521 glass Substances 0.000 claims description 9
- 229920001817 Agar Polymers 0.000 claims description 8
- 239000008272 agar Substances 0.000 claims description 8
- HVYWMOMLDIMFJA-DPAQBDIFSA-N cholesterol Chemical compound C1C=C2C[C@@H](O)CC[C@]2(C)[C@@H]2[C@@H]1[C@@H]1CC[C@H]([C@H](C)CCCC(C)C)[C@@]1(C)CC2 HVYWMOMLDIMFJA-DPAQBDIFSA-N 0.000 claims description 8
- 238000002386 leaching Methods 0.000 claims description 7
- 238000012545 processing Methods 0.000 claims description 7
- 239000000203 mixture Substances 0.000 claims description 6
- 241000209140 Triticum Species 0.000 claims description 5
- 235000021307 Triticum Nutrition 0.000 claims description 5
- 235000019743 Choline chloride Nutrition 0.000 claims description 4
- 235000019484 Rapeseed oil Nutrition 0.000 claims description 4
- 240000004808 Saccharomyces cerevisiae Species 0.000 claims description 4
- 229930006000 Sucrose Natural products 0.000 claims description 4
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 claims description 4
- XAGFODPZIPBFFR-UHFFFAOYSA-N aluminium Chemical compound [Al] XAGFODPZIPBFFR-UHFFFAOYSA-N 0.000 claims description 4
- 229910052782 aluminium Inorganic materials 0.000 claims description 4
- 239000004411 aluminium Substances 0.000 claims description 4
- 229920002678 cellulose Polymers 0.000 claims description 4
- 235000010980 cellulose Nutrition 0.000 claims description 4
- WIIZWVCIJKGZOK-RKDXNWHRSA-N chloramphenicol Chemical compound ClC(Cl)C(=O)N[C@H](CO)[C@H](O)C1=CC=C([N+]([O-])=O)C=C1 WIIZWVCIJKGZOK-RKDXNWHRSA-N 0.000 claims description 4
- 229960005091 chloramphenicol Drugs 0.000 claims description 4
- 235000012000 cholesterol Nutrition 0.000 claims description 4
- 239000012895 dilution Substances 0.000 claims description 4
- 238000010790 dilution Methods 0.000 claims description 4
- 239000012153 distilled water Substances 0.000 claims description 4
- 238000003760 magnetic stirring Methods 0.000 claims description 4
- 238000002844 melting Methods 0.000 claims description 4
- 230000008018 melting Effects 0.000 claims description 4
- WSFSSNUMVMOOMR-NJFSPNSNSA-N methanone Chemical compound O=[14CH2] WSFSSNUMVMOOMR-NJFSPNSNSA-N 0.000 claims description 4
- CHHHXKFHOYLYRE-STWYSWDKSA-M potassium sorbate Chemical class [K+].C\C=C\C=C\C([O-])=O CHHHXKFHOYLYRE-STWYSWDKSA-M 0.000 claims description 4
- 235000010241 potassium sorbate Nutrition 0.000 claims description 4
- 239000000843 powder Substances 0.000 claims description 4
- JWYIRZRIDBKTPH-UHFFFAOYSA-N propyl 2-hydroxybenzoate;sodium Chemical class [Na].CCCOC(=O)C1=CC=CC=C1O JWYIRZRIDBKTPH-UHFFFAOYSA-N 0.000 claims description 4
- 150000003839 salts Chemical class 0.000 claims description 4
- 239000000243 solution Substances 0.000 claims description 4
- 229910001220 stainless steel Inorganic materials 0.000 claims description 4
- 230000001954 sterilising effect Effects 0.000 claims description 4
- 239000005720 sucrose Substances 0.000 claims description 4
- 239000011782 vitamin Substances 0.000 claims description 4
- 229940088594 vitamin Drugs 0.000 claims description 4
- 229930003231 vitamin Natural products 0.000 claims description 4
- 235000013343 vitamin Nutrition 0.000 claims description 4
- 150000003722 vitamin derivatives Chemical class 0.000 claims description 4
- 241000426499 Chilo Species 0.000 claims description 3
- 239000003344 environmental pollutant Substances 0.000 claims description 3
- 231100000719 pollutant Toxicity 0.000 claims description 3
- 102000011632 Caseins Human genes 0.000 claims 1
- 108010076119 Caseins Proteins 0.000 claims 1
- BECPQYXYKAMYBN-UHFFFAOYSA-N casein, tech. Chemical compound NCCCCC(C(O)=O)N=C(O)C(CC(O)=O)N=C(O)C(CCC(O)=N)N=C(O)C(CC(C)C)N=C(O)C(CCC(O)=O)N=C(O)C(CC(O)=O)N=C(O)C(CCC(O)=O)N=C(O)C(C(C)O)N=C(O)C(CCC(O)=N)N=C(O)C(CCC(O)=N)N=C(O)C(CCC(O)=N)N=C(O)C(CCC(O)=O)N=C(O)C(CCC(O)=O)N=C(O)C(COP(O)(O)=O)N=C(O)C(CCC(O)=N)N=C(O)C(N)CC1=CC=CC=C1 BECPQYXYKAMYBN-UHFFFAOYSA-N 0.000 claims 1
- 229940021722 caseins Drugs 0.000 claims 1
- 235000021240 caseins Nutrition 0.000 claims 1
- 238000003745 diagnosis Methods 0.000 claims 1
- 230000036039 immunity Effects 0.000 claims 1
- 241000238631 Hexapoda Species 0.000 abstract description 7
- 238000007598 dipping method Methods 0.000 abstract description 5
- 230000000694 effects Effects 0.000 abstract description 4
- 230000007547 defect Effects 0.000 abstract description 3
- 239000002699 waste material Substances 0.000 abstract description 2
- IBSREHMXUMOFBB-JFUDTMANSA-N 5u8924t11h Chemical compound O1[C@@H](C)[C@H](O)[C@@H](OC)C[C@@H]1O[C@@H]1[C@@H](OC)C[C@H](O[C@@H]2C(=C/C[C@@H]3C[C@@H](C[C@@]4(O3)C=C[C@H](C)[C@@H](C(C)C)O4)OC(=O)[C@@H]3C=C(C)[C@@H](O)[C@H]4OC\C([C@@]34O)=C/C=C/[C@@H]2C)/C)O[C@H]1C.C1=C[C@H](C)[C@@H]([C@@H](C)CC)O[C@]11O[C@H](C\C=C(C)\[C@@H](O[C@@H]2O[C@@H](C)[C@H](O[C@@H]3O[C@@H](C)[C@H](O)[C@@H](OC)C3)[C@@H](OC)C2)[C@@H](C)\C=C\C=C/2[C@]3([C@H](C(=O)O4)C=C(C)[C@@H](O)[C@H]3OC\2)O)C[C@H]4C1 IBSREHMXUMOFBB-JFUDTMANSA-N 0.000 abstract 2
- 239000005660 Abamectin Substances 0.000 abstract 2
- 239000005886 Chlorantraniliprole Substances 0.000 abstract 2
- 229950008167 abamectin Drugs 0.000 abstract 2
- MBNMHBAJUNHZRE-UHFFFAOYSA-M thiosultap monosodium Chemical compound [Na+].OS(=O)(=O)SCC(N(C)C)CSS([O-])(=O)=O MBNMHBAJUNHZRE-UHFFFAOYSA-M 0.000 abstract 2
- 238000004166 bioassay Methods 0.000 abstract 1
- 239000000575 pesticide Substances 0.000 abstract 1
- 241000209094 Oryza Species 0.000 description 9
- 235000007164 Oryza sativa Nutrition 0.000 description 9
- 235000009566 rice Nutrition 0.000 description 9
- 238000012360 testing method Methods 0.000 description 9
- 230000009471 action Effects 0.000 description 7
- 239000002917 insecticide Substances 0.000 description 7
- 230000001018 virulence Effects 0.000 description 7
- 239000003795 chemical substances by application Substances 0.000 description 6
- 239000002574 poison Substances 0.000 description 6
- 231100000614 poison Toxicity 0.000 description 6
- 230000008859 change Effects 0.000 description 5
- 238000012544 monitoring process Methods 0.000 description 5
- 210000002784 stomach Anatomy 0.000 description 5
- 241000607479 Yersinia pestis Species 0.000 description 4
- 150000001875 compounds Chemical class 0.000 description 3
- 238000002474 experimental method Methods 0.000 description 3
- 230000002787 reinforcement Effects 0.000 description 3
- 238000011160 research Methods 0.000 description 3
- 230000009286 beneficial effect Effects 0.000 description 2
- 230000008901 benefit Effects 0.000 description 2
- 239000003153 chemical reaction reagent Substances 0.000 description 2
- 235000013305 food Nutrition 0.000 description 2
- 230000006872 improvement Effects 0.000 description 2
- 238000004519 manufacturing process Methods 0.000 description 2
- 239000000463 material Substances 0.000 description 2
- 238000011017 operating method Methods 0.000 description 2
- 210000004894 snout Anatomy 0.000 description 2
- 230000004083 survival effect Effects 0.000 description 2
- 230000000699 topical effect Effects 0.000 description 2
- 208000006877 Insect Bites and Stings Diseases 0.000 description 1
- 241000255893 Pyralidae Species 0.000 description 1
- 241000746966 Zizania Species 0.000 description 1
- 235000002636 Zizania aquatica Nutrition 0.000 description 1
- 238000003556 assay Methods 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 230000003247 decreasing effect Effects 0.000 description 1
- 238000009826 distribution Methods 0.000 description 1
- 210000005069 ears Anatomy 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 238000009472 formulation Methods 0.000 description 1
- 230000012447 hatching Effects 0.000 description 1
- 238000001802 infusion Methods 0.000 description 1
- 238000007689 inspection Methods 0.000 description 1
- 238000001990 intravenous administration Methods 0.000 description 1
- 238000011835 investigation Methods 0.000 description 1
- 230000007774 longterm Effects 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 230000008520 organization Effects 0.000 description 1
- 230000002265 prevention Effects 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 230000010076 replication Effects 0.000 description 1
- 230000012865 response to insecticide Effects 0.000 description 1
- 238000012216 screening Methods 0.000 description 1
- 238000002791 soaking Methods 0.000 description 1
- 208000024891 symptom Diseases 0.000 description 1
- 238000011285 therapeutic regimen Methods 0.000 description 1
- 238000005303 weighing Methods 0.000 description 1
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/5005—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells
- G01N33/5008—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics
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- Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Life Sciences & Earth Sciences (AREA)
- Biomedical Technology (AREA)
- Immunology (AREA)
- Hematology (AREA)
- Chemical & Material Sciences (AREA)
- Urology & Nephrology (AREA)
- Molecular Biology (AREA)
- Tropical Medicine & Parasitology (AREA)
- Medicinal Chemistry (AREA)
- Microbiology (AREA)
- Biotechnology (AREA)
- Toxicology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Food Science & Technology (AREA)
- Cell Biology (AREA)
- Physics & Mathematics (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Pathology (AREA)
- Agricultural Chemicals And Associated Chemicals (AREA)
Abstract
The invention provides a method for detecting drug resistance of chilo suppressalis, which is characterized in that a drug-carrying artificial feed sheet is prepared under the diagnostic dose of an artificial feed dipping method, and the drug resistance level of chilo suppressalis field population to drugs is rapidly detected. The pesticide is triazophos, abamectin, chlorantraniliprole and monosultap; the diagnostic dose of the medicament is that the abamectin is 1.502mg/L, the monosultap is 38.8334mg/L, the triazophos is 20.9033mg/L, and the chlorantraniliprole is 20.2148 mg/L. The method is simple and convenient to use, can fully reflect the toxicity and activity of the medicament, has small insect quantity, overcomes the defects of large insect quantity, complex operation, time and labor waste and the like of the traditional bioassay method, and can be used for quickly detecting the resistance level of the Chilo suppressalis in field population.
Description
Technical field
The present invention relates to Chemical Control of Harmful Insects field, specifically, it is related to a kind of detection drug-fast method of striped rice borer.
Background technology
Striped rice borer Chilo suppressalis (Walker), belong to lepidoptera pyralidae, are one of important pests of paddy rice,
The paddy rice that can cause harm forms the symptoms such as withered sheath, the withered heart, dead booting, dead ears and insect bite strain, and heavy losses are caused to Rice Production.In recent years
Come, due to the change of weather, cropping system and planting type, Striped Rice Borer Population is in swift and violent ascendant trend, especially 1998 with
Afterwards, it has also become the primary insect of the Yangtze river basin and south of the River rice region, the stable yields and high yield of paddy rice are seriously hindered.For a long time, change
It is to control the major measure that causes harm of striped rice borer to learn chemical control, but be due to medicament it is long-term it is unreasonable use, cause striped rice borer
Various medicaments are all generated with different degrees of resistance, a large amount of medicaments are decreased obviously to the prevention effect of striped rice borer.In order to timely
Efficiently control striped rice borer to cause harm, it is necessary to carry out monitoring for resistance to field Striped Rice Borer Population, specify Detection of insecticide resistance, formulation section
Therapeutic regimen Instructing manufacture.
Pest resistance to insecticide monitoring is the element task that pest resistance to insecticide is administered, in order to the resistance water of clear and definite insect
The flat, space-time dynamic of distribution of resistance situation, resistance change and effect of detection resistance management measure etc..It is anti-whether insect produces
The height of the property of medicine and resistance level, is mainly the LD by comparing field population and rdativery sensitive population50(or LC50) value
Judge.And whether liquefaction resistance result is accurate, then the liquefaction resistance method and sensitive Toxicity baseline with use are closely related.
Therefore, sensitive strain is cultivated, it is to carry out evil to set up accurate and easy, quick liquefaction resistance method and sensitive Toxicity baseline
The premise of worm monitoring for resistance.At present, in the monitoring for resistance of striped rice borer, different experiments room uses drop method, rice seedling in succession
Infusion process and man-made feeds medicine embrane method.Drop method is the method that FAO recommends.The worm phase of the topical application method of early stage is 6 instar larvaes,
But it is due to that 6 instar larvae feeding times are long, wastes time and energy, interindividual variation is big, and larger difficulty is brought to selection standard test worm, after
To be changed to 4 instar larvaes and establish corresponding sensitive Toxicity baseline, and Ministry of Agriculture professional standard NY/T 2058- are formulated with this
2011.But the method needs accurate drop instrument (such as micro intravenous drip instrument), and is only applicable to contact medicament, can only react and kill
The Contact toxicity of worm agent, it is impossible to embody the stomach poison function of insecticide, be unsuitable for detect stomach poison function based on medicament virulence and
Resistance.Dipping seedling method can detect stomach toxicity and action of contace poison simultaneously, fully the virulence of reflection insecticide.But, the method needs
Be prepared in advance the growth rice seedlings of 3 weeks or so, 5-6 days checkout facility results after medicine, it appears time-consuming work consuming, comparatively laborious, and
It is unfavorable on the survival of test worm so as to influence the accuracy of result of the test and rice seedling is to start to turn yellow to wither after handling 3-4 days.People
Work feed medicine embrane method can also detect stomach toxicity and action of contace poison simultaneously, can fully reflect the virulence of insecticide.But the master of the method
It is that medicament is test worm in feed surface skewness, and from newly hatched larvae to want defect, in experimental investigation result because of worm
Body is too small and is difficult to the accurate survival condition for judging test worm.Therefore, it is necessary to set up a set of easy, quick, accurate and applicability
Wide striped rice borer liquefaction resistance new method.
Diagnostic dose refers to the dosage for causing 99% sensitive population individual death.Using diagnostic dose Monitoring Insect Pest field kind
Group's resistance to the action of a drug is the method that FAO (Food and Agriculture Organization of the United Nation) is recommended.This method thinks, with after diagnostic dose processing field population, only to resist
Property individual could survive.Traditional drop method, dipping seedling method and man-made feeds medicine embrane method carries out toxicity test, cumbersome,
It is intended to set some concentration and repetition, to ensure the confidence level of result.Man-made feeds leaching medicine method is one kind of this research department invention
Detect the drug-fast new method of striped rice borer, with easy to operate, virulence that is quick, accurate and can fully reflecting various medicaments and
The advantage of activity, but do not improved still as conventional method for concentration and replication problem.But utilize the people of the present invention
The diagnostic dose of work feed leaching medicine method carries out striped rice borer field population liquefaction resistance, not only make use of man-made feeds to soak medicine method
Advantage, and under diagnostic dose, in the case where worm amount is less still can the quick detection striped rice borer resistance to the action of a drug, solve traditional poison
Power determination experiment complex steps, it is time-consuming take a lot of work and with worm amount it is big the drawbacks of, improve efficiency.
The content of the invention
It is an object of the invention to provide one kind is easy to operate, it can fully reflect medicament virulence, it is adaptable to quick detection two
Change the method for snout moth's larva field population resistance level.
In order to realize the object of the invention, the drug-fast method of striped rice borer is detected the invention provides one kind, in man-made feeds
The man-made feeds with medicine are made under the diagnostic dose for soaking medicine method, resistance level of the striped rice borer field population to medicament is detected, it is described
Medicament is Hostathion, Cupric sulfate, AVM and Rynaxypyr, the diagnostic dose of each medicament：AVM is
1.502mg/L, Cupric sulfate is 38.8334mg/L, and Hostathion is 20.9033mg/L, and Rynaxypyr is 20.2148mg/L.
The diagnostic dose of wherein each medicament is to indoor feeding by the medicament of various concentrations using man-made feeds leaching medicine method
Striped rice borer sensitive population carry out toxicity test, calculate LC99It is used as the diagnostic dose of medicament.
Further, the diagnostic dose of medicament is determined as follows：
(1) sensitive population in striped rice borer room：Raised always under conditions of not in contact with any medicament with man-made feeds indoors
Support to more than 40 generations, and the Striped Rice Borer Population that sensitiveness has screened 3 generations is screened using single pieces of an egg；
(2) dimethylformamide is used to dissolve and dilute Hostathion, AVM and Rynaxypyr for 5-7 difference
Concentration be used as toxicity test concentration；Use distilled water to dissolve and dilute Cupric sulfate to survey as virulence for 5-7 different concentration
Determine concentration；
(3) prepared by man-made feeds：
1. the preparation of A phases：135 grams of wheat germ powders, 120 grams of sucrose, 45 grams of celluloses, 155 grams of junket are added in 1000ml water
Albumen and 20 grams of dusty yeasts, are placed in automatic steam sterilizer 124 DEG C and sterilize 30 minutes after stirring；180 grams of agar are put
Enter in 2000 milliliters of water, boiled in micro-wave oven untill melting, molten agar solution will be boiled and added into the feed after above-mentioned sterilizing simultaneously
Stir；
2. the preparation of B phases：15 grams of Sodium Propyl Hydroxybenzoates, 15 Keshan potassium sorbates, 2.5 grams of chloramphenicol, 4 grams of Websters
Salt, 30 grams of reinforcement vitamin mixtures, 4 grams of Choline Chlorides, 7 milliliter of 40% formaldehyde, 10 milliliters of rape seed oils and 5 grams of cholesterol are put into
In beaker containing 1000 milliliters of aqua sterilisas, stirring and dissolving 20 minutes on magnetic stirring apparatus；
3. B phases are added when 60 DEG C are down at a temperature of A phases and are stirred, is then sub-packed in sterile aluminium box, is placed in 4
Saved backup in DEG C refrigerator；
(4) prepared by band medicine man-made feeds：Step (3) prepare man-made feeds be cut into length 1.5cm × width 1.5cm ×
Thickness 1.2mm thin slice, thin slice is chosen and pulled out after impregnating 10s in different decoctions prepared by step 2, is placed in horizontal by 45
30min is dried on the stainless steel wire wire side of degree, is sucked after unnecessary decoction, the man-made feeds thin slice with medicine is chosen into diameter 3cm
In × high 9cm flat bottom glass pipe；
(5) toxicity test：The sensitive Chilo spp larvae of the age in days of 4 age 2 is chosen into the glass with medicine man-made feeds into step (4)
In glass pipe, each concentration handles 10, carries out 4 secondary pollutants and repeats；
(6) result and diagnostic dose：Hostathion, three kinds of medicaments of Cupric sulfate and AVM 48h after chemicals treatment are examined
The dead larvae number of each concentration processing is looked into, Rynaxypyr 96h after chemicals treatment checks the dead larvae of each concentration processing
Number；Death standard：Polypide is touched with sharp tweezers, it is impossible to which normal scrambler regards as dead individuals；It is less than with compareing the death rate
10% is effectively measure, and being corrected with the control death rate, and the result to step (5) makees toxicity regression line, and calculates LC99
Go out the diagnostic dose of each medicament.
The beneficial effects of the present invention are：
1. living materials (such as wild rice stem, paddy rice or wheat) are needed in the man-made feeds that the present invention is used, with other formulas
Chilo suppressalis artificial feed it is different, the material needed for this formula can commercially buy acquisition at any time, it is ensured that testing can be with
Shi Kaizhan.
2. after man-made feeds thin slice is immersed in decoction, medicament is just sticked on man-made feeds thin slice, because larva exists
Creep and take food in man-made feeds after leaching medicine, therefore, the method can be while the stomach toxicity and contact toxicity of test agents, Ke Yichong
Divide the virulence of reflection insecticide.
3. man-made feeds can be prepared 2-4 days after completion, chemicals treatment within 2 hours can obtain result of the test, tool
The characteristics of having rapidity.
4. equipment needed for the assay method is simple, without precision instrument；Test operation is easy, and basic unit technical staff is also easy to
Application is grasped, with good practicality.
5. the application man-made feeds leaching medicine method diagnostic dose detection drug-fast method of striped rice borer, overcomes traditional sod cultivation
It is big with worm amount, it is cumbersome, time-consuming the shortcomings of take a lot of work, and can be in the case where worm amount is less, quick detection striped rice borer field
Population resistance level, specifies the resistance level of striped rice borer, instructs field medication.
Embodiment
Below by way of specific embodiment, the invention will be further described, and the corresponding experiment of combination further illustrates the present invention
Beneficial effect.But present disclosure is not limited thereto, it is impossible to therefore and it is interpreted as limitation of the scope of the invention.
The acquisition of the diagnostic dose of embodiment 1
1. research object
The present invention research object be in laboratory not in contact with conditions of any medicament with man-made feeds raise more than 40
Generation and the sensitive Striped Rice Borer Population that 3 generations have been screened using single pieces of an egg screening sensitiveness.
2. method
2.1 reagent agent：Hostathion, AVM and Rynaxypyr dimethylformamide dissolve and are diluted to 5-
7 various concentrations are standby as toxicity test concentration；Cupric sulfate distilled water dissolves and is diluted to 5-7 various concentrations as poison
It is standby that power determines concentration.
The preparation of 2.2 man-made feeds：
1. the preparation of A phases：135 grams of wheat germ powders, 120 grams of sucrose, 45 grams of celluloses, 155 grams of junket are added in 1000ml water
Albumen and 20 grams of dusty yeasts, are placed in automatic steam sterilizer 124 DEG C and sterilize 30 minutes after stirring；180 grams of agar are put
Enter in 2000 milliliters of water, boiled in micro-wave oven untill melting, molten agar solution will be boiled and added into the feed after above-mentioned sterilizing simultaneously
Stir；
2. the preparation of B phases：15 grams of Sodium Propyl Hydroxybenzoates, 15 Keshan potassium sorbates, 2.5 grams of chloramphenicol, 4 grams of Websters
Salt, 30 grams of reinforcement vitamin mixtures, 4 grams of Choline Chlorides, 7 milliliter of 40% formaldehyde, 10 milliliters of rape seed oils and 5 grams of cholesterol are put into
In beaker containing 1000 milliliters of aqua sterilisas, stirring and dissolving 20 minutes on magnetic stirring apparatus；
3. B phases are added when 60 DEG C are down at a temperature of A phases and are stirred, is then sub-packed in sterile aluminium box, is placed in 4
Saved backup in DEG C refrigerator；
It is prepared by 2.3 band medicine man-made feeds：The man-made feeds prepared in 2.2 are cut into length 1.5cm × width 1.5cm × thickness
Spend 1.2mm thin slice, thin slice choose to 2.1 prepare various concentrations decoctions in dipping 10s after pull out, be placed in horizontal by
30min is dried on 45 degree of stainless steel wire wire side, is sucked after unnecessary decoction, the man-made feeds thin slice with medicine is chosen into diameter
In the high 9cm of 3cm flat bottom glass pipe.
2.4 toxicity test：By the sensitive Chilo spp larvae of the age in days of 4 age 2 choose in 2.3 containing the glass tube with medicine man-made feeds
In, each concentration handles 10, at least carries out the repetition of 4 secondary pollutants.
2.5 inspection results and death standard：Hostathion, 3 kinds of medicaments of Cupric sulfate and AVM 48h after chemicals treatment
The dead larvae number of each concentration processing is checked, Rynaxypyr 96h after chemicals treatment checks that the larva of each concentration processing is dead
Die number.Death standard：Polypide is touched with sharp tweezers, it is impossible to which normal scrambler regards as dead individuals.It is small to compare the death rate
It is effectively measure, and being corrected with the control death rate in 10%.Data use DPS software processings, and 4 kinds of insecticides are to striped rice borer
Toxicity test the results are shown in Table 1.Calculate LC99, that is, draw the diagnostic dose of each medicament.The diagnostic dose can be used for quick detection
Resistance level of the striped rice borer field population to Hostathion, AVM, Rynaxypyr and Cupric sulfate.
Toxicity test result of the 1 four kinds of medicaments of table to striped rice borer
The diagnostic dose of medicament：AVM is 1.502mg/L, and Cupric sulfate is 38.8334mg/L, and Hostathion is
20.9033mg/L, Rynaxypyr is 20.2148mg/L.
Liquefaction resistance of the diagnostic dose of embodiment 2 to striped rice borer field population
1 detection object
This detection object is striped rice borer field to be measured population.Striped rice borer pieces of an egg are gathered from field, by the striped rice borer children of hatching
Worm, which is placed on man-made feeds, raises to be measured to 4 ages.
2 methods
Reagent agent Hostathion, AVM, Rynaxypyr and Cupric sulfate, two are carried out using diagnostic dose of the present invention
Change snout moth's larva field population Resistance detecting.
The configuration and dilution of 2.1 active compound mother liquors
Hostathion, AVM and Rynaxypyr active compound are dissolved with dimethylformamide and dilution obtains diagnosticum
Amount is standby, and it is standby that Cupric sulfate active compound obtains diagnostic dose with distilled water dissolving and dilution.
2.2 the preparation of man-made feeds：
1. the preparation of A phases：135 grams of wheat germ powders, 120 grams of sucrose, 45 grams of celluloses, 155 grams of junket are added in 1000ml water
Albumen and 20 grams of dusty yeasts, are placed in automatic steam sterilizer 124 DEG C and sterilize 30 minutes after stirring；180 grams of agar are put
Enter in 2000 milliliters of water, boiled in micro-wave oven untill melting, molten agar solution will be boiled and added into the feed after above-mentioned sterilizing simultaneously
Stir；
2. the preparation of B phases：15 grams of Sodium Propyl Hydroxybenzoates, 15 Keshan potassium sorbates, 2.5 grams of chloramphenicol, 4 grams of Websters
Salt, 30 grams of reinforcement vitamin mixtures, 4 grams of Choline Chlorides, 7 milliliter of 40% formaldehyde, 10 milliliters of rape seed oils and 5 grams of cholesterol are put into
In beaker containing 1000 milliliters of aqua sterilisas, stirring and dissolving 20 minutes on magnetic stirring apparatus；
3. B phases are added when 60 DEG C are down at a temperature of A phases and are stirred, is then sub-packed in sterile aluminium box, is placed in 4
Saved backup in DEG C refrigerator；
It is prepared by the band medicine man-made feeds of 2.3 medicaments containing diagnostic dose：The man-made feeds prepared in 2.2 are cut into length
In 1.5cm × width 1.5cm × thickness 1.2mm thin slice, the diagnostic dose decoction for the different agents for thin slice being chosen 2.1 preparations
Pulled out after dipping 10s, be placed on the stainless steel wire wire side horizontal by 45 degree and dry 30min, suck after unnecessary decoction, by band
The man-made feeds thin slice of medicine is chosen standby in diameter 3cm × high 9cm flat bottom glass pipe.
2.4 resistance diagnostics：The day old larva of 4 age 2 of field Striped Rice Borer Population is chosen to containing the band medicine people prepared in 2.3
In the glass tube of work feed, each chemicals treatment 40.Hostathion, 3 kinds of medicaments of Cupric sulfate and AVM are in chemicals treatment
48h checks dead larvae number afterwards, and Rynaxypyr 96h after chemicals treatment checks dead larvae number.Death standard：With sharp
Tweezers touch polypide, it is impossible to which normal scrambler regards as dead individuals.The population that the death rate is less than 99% is resistant population.
The man-made feeds of embodiment 3, which soak medicine diagnostic dose and the striped rice borer field population resistance to the action of a drug is determined with traditional drop method, to be compared
Compared with
Traditional drop method is, it is necessary to the drop instrument of precision, complex operation step, the multiple operating procedure per secondary check weighing, and use
Worm amount is big.Using the diagnostic dose of the present invention, band medicine man-made feeds are prepared, it is few with worm amount, and easy rapidly detection field kind
Group's resistance to the action of a drug.
Tested Striped Rice Borer Population is：Nanchang County of Jiangxi Province population, Dayu County population and Poyang County population.Topical application method
Obtain LC of 3 tested populations to 3 kinds of medicaments (Hostathion, AVM and Cupric sulfate)50, it is shown in Table 2.Simultaneously using the present invention's
Diagnostic dose 3 tested populations are carried out more than 3 kinds of medicaments resistance diagnostics, operating method such as embodiment 2, each population is in each medicine
The death rate under agent diagnostic dose is shown in Table 2.
The resistance to the action of a drug measurement result of 23 kinds of medicaments of table
(resistant multiple=field population LD50/ sensitive population LD50, sensitive population drop method LD50See Cao Mingzhang, 2004)
The death rate that man-made feeds leaching medicine diagnostic dose is determined it can be seen from data in table 2 is surveyed with traditional drop method
Surely the resistance level obtained has obvious correlation, that is, represents the diagnostic dose of the present invention and can detect striped rice borer field population
Resistance level, and the method overcome big with worm amount needed for traditional sod cultivation, the defect such as cumbersome, time-consuming is more suitable
For the detection of extensive field resistance.
Although above the present invention is described in detail with a general description of the specific embodiments, at this
On the basis of invention, some improvement can be made to it, this is obvious to those skilled in the art.Therefore, not
Deviate these improvement done on the basis of present invention spirit, belong to the scope of protection of present invention.
Claims (3)
1. one kind detection drug-fast method of striped rice borer, it is characterised in that：Tested in the case where man-made feeds soak the diagnostic dose of medicine method
The death rate of the day old larva of 4 age of striped rice borer 2, judges whether striped rice borer field population develops immunity to drugs to medicament.
2. a kind of detection drug-fast method of striped rice borer according to claim 1, it is characterised in that：The man-made feeds leaching
The diagnostic dose of medicine method is obtained by the following method：
(1) sensitive population in striped rice borer room：Indoors always under conditions of not in contact with any medicament, with man-made feeds raise to
More than 40 generations, and screen the Striped Rice Borer Population that sensitiveness has screened 3 generations using single pieces of an egg；
(2) use that dimethylformamide dissolves and dilution Hostathion, AVM and Rynaxypyr are 5-7 individual different dense
Degree is used as toxicity test concentration；Distilled water is used to dissolve and dilute Cupric sulfate dense as toxicity test for 5-7 different concentration
Degree；
(3) prepared by man-made feeds：
1. the preparation of A phases：135 grams of wheat germ powders, 120 grams of sucrose, 45 grams of celluloses, 155 grams of caseins are added in 1000ml water
With 20 grams of dusty yeasts, 124 DEG C are placed in automatic steam sterilizer after stirring and is sterilized 30 minutes；180 grams of agar are put into
In 2000 milliliters of water, boiled in micro-wave oven untill melting, molten agar solution will be boiled and add into the feed after above-mentioned sterilizing and stir
Mix uniform；
2. the preparation of B phases：15 grams of Sodium Propyl Hydroxybenzoates, 15 Keshan potassium sorbates, 2.5 grams of chloramphenicol, 4 grams of Webster salt,
Strengthen vitamin mixture, 4 grams of Choline Chlorides, 7 milliliter of 40% formaldehyde, 10 milliliters of rape seed oils and 5 grams of cholesterol for 30 grams to be put into and contain
In the beaker of 1000 milliliters of aqua sterilisas, stirring and dissolving 20 minutes on magnetic stirring apparatus；
3. B phases are added when 60 DEG C are down at a temperature of A phases and are stirred, is then sub-packed in sterile aluminium box, is placed in 4 DEG C of ice
Saved backup in case；
(4) prepared by band medicine man-made feeds：Man-made feeds prepared by step (3) are cut into length 1.5cm × width 1.5cm × thickness
1.2mm thin slice, thin slice is chosen and pulled out after impregnating 10s in different decoctions prepared by step 2, is placed in horizontal by 45 degree
30min is dried on stainless steel wire wire side, is sucked after unnecessary decoction, the man-made feeds thin slice with medicine is chosen into diameter 3cm × height
In 9cm flat bottom glass pipe；
(5) toxicity test：The sensitive Chilo spp larvae of the age in days of 4 age 2 is chosen into the glass tube with medicine man-made feeds into step (4)
In, each concentration handles 10, carries out 4 secondary pollutants and repeats；
(6) result and diagnostic dose：Hostathion, three kinds of medicaments of Cupric sulfate and AVM 48h after chemicals treatment check each
The dead larvae number of concentration processing, Rynaxypyr 96h after chemicals treatment checks the dead larvae number of each concentration processing；Extremely
Die standard：Polypide is touched with sharp tweezers, it is impossible to which normal scrambler regards as dead individuals；It is less than 10% to compare the death rate
Effectively determine, and be corrected with the control death rate, the result to step (5) makees toxicity regression line, and calculates LC99Draw each
The diagnostic dose of medicament.
3. a kind of detection drug-fast method of striped rice borer according to claim 1, it is characterised in that：The diagnosis of AVM
Dosage is 1.502mg/L, and the diagnostic dose of Cupric sulfate is 38.8334mg/L, and the diagnostic dose of Hostathion is 20.9033mg/L,
The diagnostic dose of Rynaxypyr is 20.2148mg/L.
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