抗吡虫啉棉蚜对新烟碱类药剂的交互抗性及机理的研究

山东农业大学硕士学位论文抗吡虫啉棉蚜对新烟碱类药剂的交互抗性及机理的研究姓名：史晓斌申请学位级别：硕士专业：农药学指导教师：王开运2011-06-1011.60%~70%4541.7F0~F9F12~F36F36~F4538.281414.414.4940.32.5.784.523.681.201.061.033.-S-64.01%49.31%36.20%-S-32.29%32.47%26.45%-S-18.00%5.71%18.28%TPPPBO2.321.981.311.67DEM1.074.LC2043.7%23.4%2AbstractInordertoconfirmthecross-resistancetoneonicotinoidinsecticides,resistancemechanismandbiologicalcharacteristicsofimidacloprid-resistantcottonaphid,andtoguidetherationaluseofneonicotinoidinsecticides,thedippingmethod,microspotmethod,biochemicalmethodandsystematicobservationmethodwereadoptedwiththestrainsoftheimidacloprid-resistantpopulation,thexiajinpopulationandthesusceptiblepopulation.Thetrendsofresistancedevelopmentwereresearched,atthesametimethecross-resistancetoneonicotinoidinsecticides,resistancemechanismandbiologicalcharacteristicsofimidacloprid-resistantcottonaphidwereinvestigated.Theresultswereasfollows:1.TheresistancelevelofA.gossypiitoimidaclopridreachedtomediumresistancewith41.7-foldafterselectionof45generationwiththepressuretokill60%to70%individualsofthepopulation.TheresistanceofA.gossypiitoimidaclopridincreasedslowlyatthefirst9generations,andthenincreasedrapidlyfromthe12thtothe36thgenatation,atlasttheresistanceincreasedslowlyagainfromthe36thtothe45thgenatation.Thelevelofresistancewillcontinuetodevelopmentiftheselectioniskepton.Whentheresistanceratiosreachedto38.2-fold,theresistancestabilityofA.gossypiitoimidaclopridwasdetermined.Theresistancereducedrapidlyinthefirst8generation,andtheresistancegraduallykeptsteadyto14.4-foldafter14generation.Theimidacloprid-resistantcottonaphidof14.4-foldwasselectedoveragain,theresistanceofA.gossypiiincreasedrapidly,andtheresistanceincreasedto40.3-foldafter9genetarions.2.BasedonLC50values,thetoxicityratiosofnitenpyram,acetamipridandthiaclopridwere5.78-,4.52-and3.68-fold,respectively,suggestingcross-resistancebetweenimidaclopridandtheseneonicotinoids.However,therewasnocross-resistanceexistedbetweenimidaclopridandthreeneonicotinoidinsecticidesincludingclothianidin,thiamethoxamanddinotefuran.3.ThebiochemicalresultsshowedthatdinotefurancouldsignificantlyinhibittheCarE,GSTandAChEoftheimidacloprid-resistantpopulation,withtheinhibitionof64.01%,49.31%and36.30%respectively.DinotefuranalsoshowedobviousinhibitionontheCarE,GSTandAChEoftheXiajinmulti-resistantpopulation,withtheinhibitionof32.39%,32.47%and26.45%.NitenpyramshowedlittleeffectontheCarE,GSTandAChEofthe3imidacloprid-resistantpopulation,withtheinhibitionof18.00%,5.71%and18.28%.TPPandPBOhadobviouseffectonthesynergytoimidaclopridandnitenpyram,withthesynergismratioof2.32-and2.98-foldofimadacloprid,whilelittleeffectonthesynergytodinotefuran,withthesynergismratioof1.31-and1.67-fold.DEMshowednoobvioussynergismtothethreeinsecticideswiththesynergismratioof1.07-fold.4.Inthestudyofsublethaleffects,imidaclopridatLC20dosescouldsuppressweightgainandhoneydewexcretion,withtheinhibitionof43.7%and23.4%,resprcitively,butshowednosignificanteffectsonlongevityandfecundityoftheimidacloprid-resistantcottonaphid.However,otherneonicotinoidinsecticidesshowedsignificantadverseeffectsonbiologicalcharacteristics(bodyweight,honeydewexcretion,longevityandfecundity)intheorderofdinotefuranthiamethoxamandclothianidinnitenpyramthiaclopridandacetamiprid.Keywords:Aphisgossypii;cross-resistance;neonicotinoids;sublethalconcentrations;biochemicalmechanismhHourminMinutesSecondLD50MedianlethaldoseLC50MedianlethalconcentrationmLMilliliterCentigradeµg10-6g,MicrogrammgMilligrammegGrammeµL10-6L,MicrolitreLLiliterr·min-1Rotate·minute-1AChEAcetylcholinesteraseGSTGlutathione-S-transferase–S–CarECarboxylesterasesPrProteinODOptialdensityPBOPiperonylbutoxideTPPTriphenylphosphateDEMDiethylmaleateµmol·L-110-6mole·L-1UVUltra-violetUActiveUnitEDTAEthylenediaminetetraaceticacidKdrKnock-downresistance41.Aphisgossypii(Glover)200020041999199119921.11.1.12050701600019971964105916051482319641965150.8588.8625.0715.7819601988500~7001994196314823197714008001987405(19901996)FurkC(1990)801985LD50309411836519.7401.2198719914146.3419981997199575127.918341.253122.452000889.1480198010000~1200080001983600019843,00019851000~20001987808090199019901997767.86553.5LD501987344.767.81997199561996199819945.362.251.8319952/52000Sunetal,19941.1.2Han,19981.1.2.1IRAC-online1.1.2.2P450-S-(GST)1P450P450P450P4501965RayP450P4507P450(PBO)(1993)P450(1999)(2000)PBOP450(1998)(2003)2(Villatteetal.1999)Wang2002α-NAα-NA3.803.02(2003)(19981995)Weichel(2003)Sun2005α-NAα-NBCarENewcomb(1997)E3LcαE7Newcomb(1997)LcαE7G137D8()W251L/S(100)(Heidarietal2004)mRNAZhu(1999)(Anisopteromaluscalandrae)mRNA303-S--S-(GSTs)(GSH)GSHGSTsmRNAGSTs(2005)GST(1989)Prapanthadara(1995)DDTGST(1996)GSTGSTP450Konus(2008)GSTCYP6B61.1.2.3kdrAChE2003Mutero(1994)SaltilloBygdeaPierrefeu45F115SI199VI199YG303AF368YWalsh(2001)45V180(150220)LG262(227303)AG262(227303)VF327(290368)YG365(328406)A2Weill(2003)G119S799Zhu(1996)S291G(2002)A585TMutero(1994)Walsh(2001)Kdr19982003RT-PCR1986DDT19951.1.31.1.3.1(1988)181616.54324.622.3101.1.3.21.1.3.360801-21.21.2.11.2.1.1200049.62000Ap