B作物的昆蟲抗藥性管理

InsectResistanceManagementforBtCrops

Bt作物的昆蟲抗藥性管理TessaMilofsky,M.S.andSharleneMatten,Ph.D.泰莎.米洛夫斯基碩士沙利恩.馬頓博士U.S.EnvironmentalProtectionAgencyOfficeofPesticidePrograms美國環(huán)保局殺蟲劑項(xiàng)目辦公室1IRM:whatisit?

昆蟲抗藥性管理定義

Insectresistancemanagement(IRM)isastrategytomitigatethedevelopmentofinsectsresistanttoapesticide(e.g.,resistancetoBacillusthuringiensisorBt)

昆蟲抗藥性管理(IRM)是為了減輕害蟲對殺蟲劑(例如對BT)產(chǎn)生抗性而采取的一種策略.2IRM:whyisitimportant?

IRM的重要性MaintainingtheefficacyofBt,alow-riskpesticide,isbeneficialforthefollowingreasons:維持低風(fēng)險殺蟲劑Bt的功效有如下的優(yōu)點(diǎn):BtreplaceshigherriskpesticidesBt殺蟲劑可以代替其他高風(fēng)險性的殺蟲劑Lessexposureofnontargetorganismstosuperficiallyappliedpesticides降低非目標(biāo)生物暴露于殺蟲劑的可能性UseofBtcropsmayprovidegreaterprofitstofarmers,duetoreduceduseofinsecticides由于減少了殺蟲劑的使用,Bt作物可以給農(nóng)民帶來更大的收益3CriticalcomponentsofIRMplan

IRM計(jì)劃的重要組成成分Pestbiology害蟲生物學(xué)Dose劑量Cross-resistance抗性轉(zhuǎn)移Refuge避難所Modeling建立模型Resistancemonitoring抗性監(jiān)測Remedialaction矯正措施Complianceassurance確保規(guī)范性Growereducation種植者教育4Pestbiology

害蟲生物學(xué)Importantfactorsinclude:NumberofinsectgenerationsperyearHostrangeInsectmovementanddispersalpatternsMatingandovipositionalbehaviorPopulationdynamics要點(diǎn)包括:每年害蟲繁殖的代數(shù)宿主的范圍害蟲移動和分布的模式交配和產(chǎn)卵行為種群動態(tài)5Pestbiology:anexample

害蟲生物學(xué)舉例Europeancornborer(ECB),Ostrinianubilalis1-4generationsperyearPolyphagous(preferscorn)Movement:Larvae-canmovefromplant-to-plantwithinafieldAdults-mostlylocalmovement,butsomelongdistancemigratorymovementcanoccur歐洲玉米螟(ECB)每年繁殖1-4代雜食性(偏好玉米)移動幼蟲可以在同一田地內(nèi)不同植株上移動成蟲大部分在本地活動,但也有可能進(jìn)行長距離遷移Female雌性Male雄性lateinstars晚期幼蟲ECB6Dose劑量Dose=theamountoftoxinexpressedovertimebyatransgeniccrop,relativetothesusceptibilityofatargetpest;canbedeterminedthroughfieldefficacytrialsorlaboratorybioassays劑量=轉(zhuǎn)基因作物在一定時間內(nèi)表達(dá)的毒素的量,和目標(biāo)害蟲的敏感性相關(guān);通常由田間功效實(shí)驗(yàn)或?qū)嶒?yàn)室生物測試所決定.Highdose=atleast25xtheamountoftoxinneededtokillallsusceptibletargetinsects;preferredoverlowdoseeventsforrefuge-basedIRMstrategies高劑量=至少為殺死所有敏感目標(biāo)害蟲劑量的25倍;在以避難所為基礎(chǔ)的昆蟲抗性管理策略中，使用高劑量遺傳事件體要比低劑量遺傳事件體好7Doseassumptions

劑量假設(shè)Threegenotypesforresistance/susceptibility(SS,rS,rr)Resistance(rr)isrecessiveandrareRefugesuppliesSS(homozygoussusceptible)genotypeRandommating:rrxSS→rSrSxSS→rSorSSSSandrSsusceptibletohighdose三種抗性/敏感性基因型(SS,rS,rr)抗性基因(rr)是隱性的,少見的安全基因型SS(純合敏感)隨機(jī)交配:rrxSS→rSrSxSS→rSorSSSS和rS對高劑量敏感LC500100低高8Doselevels:someexamples

關(guān)于劑量水平的一些例子Lepidopteran-activecorn(Cry1Ab&Cry1F):HighdoseEuropeancornborer(ECB),OstrinianubilalisSouthwesterncornborer(SWCB)DiatraeacrambidoidesNon-highdoseCornearworm(CEW),Helicoverpazea鱗翅目敏感玉米(Cry1Ab&Cry1F):高劑量歐洲玉米螟(ECB),Ostrinianubilalis西南玉米螟(SWCB)Diatraeacrambidoides非高劑量玉米棉鈴蟲(CEW),HelicoverpazeaWCRfemalemalelateinstarsECB9Doselevels:someexamples

關(guān)于劑量水平的一些例子Rootworm-activecorn(Cry3Bb1,Cry34/35,mCry3A)Non-highdoseWesterncornrootworm(WCR),DiabroticavirgiferavirgiferaNortherncornrootworm(NCR),D.barberiMexicancornrootworm(MCR),D.virgiferazeaSecondarypestsNon-highdoseforsomepests根蟲敏感玉米(Cry3Bb1,Cry34/35,mCry3A)非高劑量西部玉米根蟲(WCR),Diabroticavirgiferavirgifera北部玉米根蟲(NCR),D.barberi墨西哥玉米根蟲(MCR),D.virgiferazea次級害蟲非高劑量WCRfemalemalelateinstarsECB10Cross-resistance

抗性轉(zhuǎn)移WhenresistancetoasingleBttoxinconferspestresistancetoaclusterofrelatedtoxinsFactorsaffectingcross-resistanceinclude:Similarity/dissimilarityofcommercializedtoxinsUseofsingletoxinversusmultipletoxinsLevelofresistanceinaninsectpopulation害蟲對單一Bt蛋白的抗性使得害蟲對其他一系列相關(guān)毒素都產(chǎn)生抗性影響抗性轉(zhuǎn)移的因素包括:商品用毒素的異同點(diǎn)單一毒素和復(fù)合毒素的使用昆蟲種群的抗性水平11Cross-resistance:anexample(抗性轉(zhuǎn)移:舉例說明)BindingsitediagramforEuropeancornborer(歐洲玉米螟結(jié)合位點(diǎn)簡圖)Cry1AbCry1FCry1Ac220KDaCadherin154KDaAPN145/167KDaAPNOtherEuropeancornborer(basedonHua,G.,Masson,L.,Jurat-Fuentes,J.L.,Schwab,G.,Adang,M.J.2001.BindinganalysesofBacillusthuringiensisCryd-endotoxinsusingbrushbordermembranevesiclesofOstrinianubilalis.Appl.Env.Microbiol.67:872-879)(其他)12Refuge避難所AnyhostplantthatdoesnotproduceBttoxinandhasnotbeentreatedwithconventionalBtformulations(maybetreatedwithnon-Btinsecticide)任何不產(chǎn)生Bt毒素并且也未經(jīng)常規(guī)Bt殺蟲劑處理過的宿主植物(可能經(jīng)非Bt殺蟲劑處理過)Enablesmatingbetweenresistantandsusceptibleadults,resultinginproductionofsusceptibleoffspring抗性和敏感性成熟植株雜交可以產(chǎn)生敏感性的后代13Refugeoptions 避難所的選擇Structuredrefuge=specifiedsize,locationinrelationtoBtfield,andcrop/plantcompositionCommunity-basedrefuge=neighboringgrowersestablishsharedstructuredrefugeUnstructuredornaturalrefuge=useofalternativehostplants/cropsinplaceofdefinedrefuge結(jié)構(gòu)避難所=特定大小,并且位置和Bt植物種植區(qū)有一定關(guān)系的區(qū)域,此區(qū)域既有農(nóng)作物,又有植被以社區(qū)為基礎(chǔ)的避難所=相鄰的種植者建立的一種共享的結(jié)構(gòu)避難所非結(jié)構(gòu)或稱天然避難所=用一種寄主植物和作物互相交替種植的模式來代替人為限定的避難所模式14Refugeoptions 避難所的選擇Seedbagmix=BtseedbagscontainBtseedandnon-Btseed(e.g.,80%Bt,20%non-Bt);seedmaybebaggedseparatelyormixedtogetherRegionallimit=national/regionalgovernmentenforcedlimitonBtseedsales(e.g.,80%indefinedregion)混裝種袋=在這種含Bt種子的袋子中既含有Bt種子,也含有非Bt種子(例如:80%Bt種子,20%非Bt種子),種子可以先按小袋分裝好再混和,也可以直接混在一起.區(qū)域限制=國家或地方政府對Bt種子的買賣進(jìn)行嚴(yán)格限制(例如:在一定區(qū)域內(nèi)，只允許80%的Bt種子的買賣)15SeedmixBlockIn-fieldstrips田地內(nèi)的條帶模式Internalblock(s)內(nèi)部種植模式Border(block)周邊種植模式Structuredrefugeoptions

結(jié)構(gòu)避難所的選擇Externalblock外部種植模式種子混和16Otherrefugeoptions

其他避難所選擇Communityrefuge社區(qū)避難所模式80%Bt

seedcorn玉米種子20%non-Bt

seedcorn玉米種子Naturalrefuge天然避難所BtcottonBt棉花w/TBWSoybean大豆Tobacco煙草Tobaccobudworm,煙草蚜蟲Heliothisvirescens

17Btcornrefugespecifications:anexample

Bt玉米避難所規(guī)范示例20%refugerequiredincornbelt;50%refugerequiredincorn/cottonregionStructuredrefugeisrequired.Refugesmaybeplantedinoneofthefollowingdesigns:In-fieldstrips(4-6rows)Blocks(internalorexternal)Refugerequirementsarethesameforlepidopteranandcoleopteranactiveproducts在Bt玉米田中20%為避難所,在玉米/棉花田中則為50%結(jié)構(gòu)性避難所是必須的.避難所可以按如下的設(shè)計(jì)來耕作:田地內(nèi)條帶(4-6行)內(nèi)部或外部田塊18Modeling模擬Usefultoolforpredictingthelikelihoodofresistance,forevaluatingthedurabilityofIRMstrategies,andforhelpingtofocusdataneeds.Modelcomponentsinclude:Biologicalinformation(pestbiology,populationdynamics)Genetics(R-alleledominance/frequency,crossresistance)Operationalaspects(dose,single/stackedtoxin(s),refuge)建立模型是預(yù)測抗藥可能性,評價IRM策略持久性和幫助調(diào)整數(shù)據(jù)需求的一種有效方法.模型包括以下組成成分:生物信息(害蟲生物學(xué),種群動態(tài))遺傳學(xué)(R-等位基因顯性/頻率,抗性轉(zhuǎn)移)運(yùn)行情況(劑量,單一的/疊加的毒劑,避難所)19Resistancemonitoring

抗性監(jiān)測CanhelptoprolongtheefficacyofaninsectresistantcropDesignedtodetectresistance2-3yearsbeforefieldfailureTargetpestsamplingconsistsof:Growerreportsofunexpectedpestdamageininsect-protectedcropfieldsRandompopulationsampling:SamplingbasedonpestprevalenceinregionSamplingtarget:numberofinsectsneededtodetermineresistanceatp<0.01

可以延長抗蟲作物的功效可以在失效前監(jiān)測抗性2-3年目標(biāo)害蟲取樣包括:種植者在抗蟲作物區(qū)域內(nèi)由害蟲造成的意外損失報告隨機(jī)種群取樣:害蟲流行區(qū)域取樣取樣目標(biāo):用來確定抗性所需的害蟲數(shù)量(p<0.01)20Resi德stan眠cem億onit湖orin儀g(c沿ont.繪)抗性監(jiān)捷測(續(xù)結(jié))Bio禿ass聚ays職ar霞eu慎sed鮮to租de六tec致tp擇ote致nti夜al描res蛋ist則anc磚ei伐ns味amp絕led惡po惹pul刊ati虹ons匹.A輝ssa左ys枝inc合lud撇e:Pes撇ts蔑usc就ept淺ibi趴lit宿y(咽LC50)Dis晌cri只min道ati材ng它Dos煩e(貪LC99)F2scre摸enCon蘋fir符mat銀ion命st志udi晉es鞏are顛us獲ed親to頂ver趨ify綱re燭sis秀tan智ce.石Qu扎est絹ion討si榨ncl房誠ude毛:Ist稅here碑fie所ldf挎ailu境re?Is軍the渠in姜sec莫tr鞋esi皺sta瓣nce潑tr泛ait旁ex孔pre暑sse堅(jiān)d?Isi甜nsec艘tre膜sist賣ance倡tra嗚iti也nher止ited躍?用生物宿鑒定來警檢測樣俱品種群武中的潛桂在抗性炮.生吩物鑒定鮮包括:害蟲敏感醬性鑒定(LC50)差異劑量枕(LC99)F2篩選確認(rèn)研究欲用來核實(shí)客抗性.包款括:區(qū)域失效決了么?抗性特征艘表達(dá)了么孫?抗性性可禍遺傳么?21Resi盯stan鉛cem熊onit危orin蠻g:p傲osit站ive絕hits捧forBtcor裝ni感nt嬌he坑US?抗性監(jiān)測躺:美國Bt玉米正采克樣數(shù)In箏200妨1,廊mor等tal坐ity潔fo殊ro恭ne靜Eur辰ope敢an旺cor纖nb癥ore閣r(輔ECB貍)p法opu基lat塵ion羞(K裂a(bǔ)nd纏iyo姻hi,趨MN禁)w御as言<9絲式9%搭in哄Cry戰(zhàn)1Ab伴di勸scr老imi綠nat勞ing制do增se或ass升ayFoll巾ow-u告pte樂sts乖withBtplan濁t(yī)ma耳teri態(tài)alr朱evea驢led洋that通pop塵ulat開ion牙was煩not婆fiel得dre名sist鄙antNofiel萄dre辟sist尿anceconf柳irme腔dfo勇ran咬ymo哥nito址redBtcor蓬np救est2001年在Cry1只Ab不同劑量吃分析中,粥歐洲玉卡米螟種群呀的死亡率括小于99息%Bt作物的允后續(xù)測穿試表明鼠種群并盈沒有田灶間抗藥團(tuán)性被監(jiān)測胃的Bt玉米害蟲能沒有被證頃實(shí)產(chǎn)生田踐間抗藥性22Reme混dial枝act窯ion魚plan矯正措嚷施If徹res以ist耕anc館ei明sc蝴onf鍋irm艙ed酸t(yī)hr獻(xiàn)oug嬸hr雨esi逐sta注nce堂mo咬nit疑ori例ng,貨a正re餐med揭ial釘ac砌tio綢np駐lan懼is動im下ple廳men陷ted嚷to歌co值nta場in青or秀slo圾wt幅he專spr丸ead粘of很re錦sis勵tan物tp三est逼po寫pul鏈ati遍ons如果抗性園監(jiān)測確認(rèn)難了抗藥性承,矯正患措施可以例減緩害蟲夾種群抗藥填性的傳播23Rem桂edi股al捎act拳ion夢pl源an:抵an構(gòu)ex儀amp律le矯正措施鑄舉例EPA’鋒spl殖ani可nclu忌des:Mand柿ator民yre趁port穩(wěn)ing扛ofs躍uspe躍cted痛res紡ista優(yōu)nceUse疤of雖al郊ter堡nat油ec略ont隆rol妄me多asu嗽res夢in敞af歌fec怪ted鑒ar沒easInc凡rea譽(yù)sed薦re常sis托tan租ce刺mon迅ito語rin壘gSus棚pen仿sio秒no張fBtcro拔ps坑ale批si色na灣ffe源cte雞da敢rea蘋sInv顫est呆iga綿tio漆no粱fc喇a(chǎn)us錄eo惹fr次esi測sta炸nce美國環(huán)保憲局措施包孕括:強(qiáng)制要石求對有稿懷疑的爐抗藥性鎮(zhèn)表現(xiàn)上纏報在有影煌響區(qū)域膠使用其縱他管理已措施增強(qiáng)抗性白監(jiān)測在被影響臟區(qū)域終止Bt玉米銷翼售對抗藥性坑原因進(jìn)行奶調(diào)查24Com傭pli旗anc還ea付ssu來ran悅ce確保規(guī)塵范性Com肢pli揪anc考em深eas更ure鄉(xiāng)豐si界mpl萍eme孔nte在dt睡oe框nsu倘re礎(chǔ)tha潮tg吐row課ers龜co命mpl泰yw疾ith挺ap索pro脫ved否IR勻Mp朗rog吳ram規(guī)范操作師可以確保糟種植者按云照害蟲抗詞藥性管理球規(guī)則執(zhí)行烈耕種操作時.25Com西pli柏anc籠ea催ssu要ran品ce:芝an箏ex透amp柿le確保規(guī)玻范性舉令例EPA猾re司qui煎res富re蓄gis落tra茄nts透to晶im薦ple番men區(qū)tt評he莊fol乏low歡ing制me象asu充res劫:Cont持ract尖saf模firm缸ing梳grow憲era搶gree京ment壇to鐘comp輕lyw包ith扭appr詢oved游IRM店str粒ateg桶y3rdpart款ygr鍵ower勸sur軍veys皇of胡IRM拜impl遙emen賤tati央onOn-f尺arm振fiel吐dvi聽sits挑to懂veri反fyI沉RMi堪mple鈴ment鬼atio蕉nTips投and懷com踩plai誕nts耕onn拴on-c矮ompl歐iersSusp始ensi粒ono謙fse桌eda框cces騎sfo絕rno賽n-co穗mpli拴ers美國環(huán)保贊局規(guī)定登胃記者必須盾實(shí)施以下射方法:合同規(guī)糕定種植描者必須擁遵守IRM策略第三方對IRM執(zhí)行情況煮進(jìn)行調(diào)查田間觀慕察以確匆定IRM的執(zhí)行情俘況揭發(fā)和殖投訴不消遵守規(guī)乳則的種竿植者對不遵守察規(guī)則的種蔬植者終止遼種子供應(yīng)26Grow吊ere共duca拾tion種植者教槳育Educ掙atio河nab逮out擔(dān)the霜need菠for劃IRM法,ho吩wto廊imp辱leme帽ntr瘦equi箏reme膽nts,領(lǐng)and道gro碑wer飼resp糞onsi情bili停tyi翁nco鞭mply嘩ing竊with尊IRM盤req業(yè)uire脂ment她s對IRM策略的必涌要性,域怎樣完成捷要求,墊和種植者蜻對完成IRM要求所泊擔(dān)負(fù)的手責(zé)任進(jìn)埋行教育27Summ拿ary撐ofE衣PA’s撫IRM麗req盟uire怒ment希sfo躍rle顆pido烈pter府a(chǎn)n-a褲ctiv功eBtcorn美國環(huán)胞保局對棋鱗翅目席昆蟲敏餐感的Bt玉米IRM要求概淡述20%refu奇gei爆nco奔rnb鴨elt;佛50%愁ref舍uge島inc體otto彼nbe碌ltStru抱ctur敲edr喂efug狗ere動quir線ed.撓Refu攀ges斥plan鏡ted怕ino飼neo功fth暫efo搏llow律ing物desi柿gns:in-f顆ield粗str找ips昆(4-6校row震s)bloc董ks(世inte賠rnal客or是exte醒rnal耗)Ind沿ust叉ry-紀(jì)wid氧ec漫oll噴abo酬rat鵝ion白o(hù)n哄co捷mpl違ian堵ce舊sur衣vey茶s在玉米種嶺植帶中2徐0%為避各難所;道在棉花種陜植帶中為部50%結(jié)構(gòu)性征避難所舞是必需流的.避端難所種罵植需為冰如下設(shè)惹計(jì)中的訂一種:田間條良帶(4慌-6拘行)田塊(內(nèi)鵝部或外部秤)廣泛合聯(lián)作開展閥規(guī)范性滔調(diào)查28Summ綱ary由ofE海PA’s時IRM羞req揪uire褲ment加sfo撫rle棕pido塌pter帝an-a撿ctiv筒eBtcorn漂(co及nt.)對鱗翅目位昆蟲敏感鎖的Bt玉米IRM要求概述畝(續(xù))Ind偶ust導(dǎo)ry-杏wid譜eu脹nif舊orm磨ity僑of卷:Gro睬wer輝ag喝ree靜men懲tsGrow傘ert叨rain辱ingImpl頭emen男tati禾ono例fti燥psa歉ndc便ompl朝aint忠s以下行痰為保持陷高度一掙致性:種植者誼協(xié)議種植者培途訓(xùn)監(jiān)測和球投訴的浙實(shí)施29Sum平mar熄yo貌fE緩PA’候sI族RM熔req擱uir攜eme起ntsfor挖lepi壁dopt榮eran睡-act料iveBtcorn悉(co辭nt.)對鱗翅目辮昆蟲敏感寒的Bt玉米IRM要求概凳述(續(xù)庭)Annu展alr能epor寸ting例req盛uire志ment搞s:－Com梨pli艷anc鳴ea涂ssu陳ran崇ce叫(e.g既.,surv森eys,店gro悅wer忽cont桿ract施s)－Chan傲ges匯tog衣rowe揪red氣ucat恢ion－Res扇ist公anc稠em午oni辨tor淘ing朗da憑ta－Sal鼠es傻dat累a年度報告姥需要:規(guī)范性確替認(rèn)材料(食例如調(diào)查壩,種植者功合同)種植者教章育的變化抗性監(jiān)測盤數(shù)據(jù)銷售數(shù)據(jù)30Data鄭sou仰rces憲for恰EPA仇’sI步RMp破rogr秤am美國環(huán)保狐局IRM項(xiàng)目的貧數(shù)據(jù)來抵源Pub勵lis西hed坦sc殖ien先tif疼ic它lit節(jié)era費(fèi)tur直eIndu刃stry洞sub狹miss滲ionsExpe傍rts恰(aca覽demi撕a,g徐over餅nmen刻t,i配ndus退try)Mee頓tin者gs此(ad彼vis辣ory痛pa繪nel塘s,柔wor疊ksh伏ops吹,E閥SA,說NC遞205介,N停C46礦)Fiel標(biāo)dvi菠sits已出版的旨科學(xué)文獻(xiàn)工業(yè)企靜業(yè)的報觸告專家(學(xué)泰術(shù)界,政們府,工業(yè)陷界)會議(顧哲問團(tuán),研違討會,ESA,貧NC20馬5,NC誤46)田間觀膨察31Keys徐to沫succ剝ess頑atE拒PA美國環(huán)畜保局(EPA辰)取得成滑功的關(guān)姑鍵EPA訓(xùn)isr奴espo子nsiv壞eto贊new助sci旅enti招fic土info洲r(nóng)mat忙ionEPA增se狐eks某sc積ien涂tif撞ic雖inp胖ut地fro翻ms州tat扣ea愉nd謙fed禾era港la愈gen逝cie纖s,破aca氧dem轎ic逐exp絕ert擺s,糊and取pu劍bli盲ci爆nte資res冤tg獻(xiàn)rou亭psEPA對最新的恐科技信息蓄反應(yīng)積極EPA從各州后和聯(lián)邦粥的政府戲部門,饞學(xué)術(shù)專周家和公聚眾團(tuán)體現(xiàn)中尋求豎科技力庸量32Keys問to篇succ售ess席atE特PA美國環(hu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