B作物的昆蟲(chóng)抗藥性管理

InsectResistanceManagementforBtCrops

Bt作物的昆蟲(chóng)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(guó)環(huán)保局殺蟲(chóng)劑項(xiàng)目辦公室1IRM:whatisit?

昆蟲(chóng)抗藥性管理定義

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

昆蟲(chóng)抗藥性管理(IRM)是為了減輕害蟲(chóng)對(duì)殺蟲(chóng)劑(例如對(duì)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(xiǎn)殺蟲(chóng)劑Bt的功效有如下的優(yōu)點(diǎn):BtreplaceshigherriskpesticidesBt殺蟲(chóng)劑可以代替其他高風(fēng)險(xiǎn)性的殺蟲(chóng)劑Lessexposureofnontargetorganismstosuperficiallyappliedpesticides降低非目標(biāo)生物暴露于殺蟲(chóng)劑的可能性UseofBtcropsmayprovidegreaterprofitstofarmers,duetoreduceduseofinsecticides由于減少了殺蟲(chóng)劑的使用,Bt作物可以給農(nóng)民帶來(lái)更大的收益3CriticalcomponentsofIRMplan

IRM計(jì)劃的重要組成成分Pestbiology害蟲(chóng)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(cè)Remedialaction矯正措施Complianceassurance確保規(guī)范性Growereducation種植者教育4Pestbiology

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

害蟲(chóng)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(dòng)幼蟲(chóng)可以在同一田地內(nèi)不同植株上移動(dòng)成蟲(chóng)大部分在本地活動(dòng),但也有可能進(jìn)行長(zhǎng)距離遷移Female雌性Male雄性lateinstars晚期幼蟲(chóng)ECB6Dose劑量Dose=theamountoftoxinexpressedovertimebyatransgeniccrop,relativetothesusceptibilityofatargetpest;canbedeterminedthroughfieldefficacytrialsorlaboratorybioassays劑量=轉(zhuǎn)基因作物在一定時(shí)間內(nèi)表達(dá)的毒素的量,和目標(biāo)害蟲(chóng)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(cè)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(chóng)劑量的25倍;在以避難所為基礎(chǔ)的昆蟲(chóng)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(jiàn)的安全基因型SS(純合敏感)隨機(jī)交配:rrxSS→rSrSxSS→rSorSSSS和rS對(duì)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(chóng)(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(chóng)敏感玉米(Cry3Bb1,Cry34/35,mCry3A)非高劑量西部玉米根蟲(chóng)(WCR),Diabroticavirgiferavirgifera北部玉米根蟲(chóng)(NCR),D.barberi墨西哥玉米根蟲(chóng)(MCR),D.virgiferazea次級(jí)害蟲(chóng)非高劑量WCRfemalemalelateinstarsECB10Cross-resistance

抗性轉(zhuǎn)移WhenresistancetoasingleBttoxinconferspestresistancetoaclusterofrelatedtoxinsFactorsaffectingcross-resistanceinclude:Similarity/dissimilarityofcommercializedtoxinsUseofsingletoxinversusmultipletoxinsLevelofresistanceinaninsectpopulation害蟲(chóng)對(duì)單一Bt蛋白的抗性使得害蟲(chóng)對(duì)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(chóng)種群的抗性水平11Cross-resistance:anexample(抗性轉(zhuǎn)移:舉例說(shuō)明)BindingsitediagramforEuropeancornborer(歐洲玉米螟結(jié)合位點(diǎn)簡(jiǎn)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(chóng)劑處理過(guò)的宿主植物(可能經(jīng)非Bt殺蟲(chóng)劑處理過(guò))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(lái)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(guó)家或地方政府對(duì)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(chóng)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(lái)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(cè)抗藥可能性,評(píng)價(jià)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(chóng)生物學(xué),種群動(dòng)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(cè)CanhelptoprolongtheefficacyofaninsectresistantcropDesignedtodetectresistance2-3yearsbeforefieldfailureTargetpestsamplingconsistsof:Growerreportsofunexpectedpestdamageininsect-protectedcropfieldsRandompopulationsampling:SamplingbasedonpestprevalenceinregionSamplingtarget:numberofinsectsneededtodetermineresistanceatp<0.01

可以延長(zhǎng)抗蟲(chóng)作物的功效可以在失效前監(jiān)測(cè)抗性2-3年目標(biāo)害蟲(chóng)取樣包括:種植者在抗蟲(chóng)作物區(qū)域內(nèi)由害蟲(chóng)造成的意外損失報(bào)告隨機(jī)種群取樣:害蟲(chóng)流行區(qū)域取樣取樣目標(biāo):用來(lái)確定抗性所需的害蟲(chóng)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(cè)(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(chéng)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(lái)警檢測(cè)樣俱品種群武中的潛桂在抗性炮.生吩物鑒定鮮包括:害蟲(chóng)敏感醬性鑒定(LC50)差異劑量枕(LC99)F2篩選確認(rèn)研究欲用來(lái)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(cè)躺:美國(guó)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(lè)sts乖withBtplan濁t(yī)ma耳teri態(tài)alr朱evea驢led洋that通pop塵ulat開(kāi)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(cè)穿試表明鼠種群并盈沒(méi)有田灶間抗藥團(tuán)性被監(jiān)測(cè)胃的Bt玉米害蟲(chóng)能沒(méi)有被證頃實(shí)產(chǎn)生田踐間抗藥性22Reme混dial枝act窯ion魚(yú)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(dòng)im下ple廳men陷ted嚷to歌co值nta場(chǎng)in青or秀slo圾wt幅he專spr丸ead粘of很re錦sis勵(lì)tan物tp三est逼po寫pul鏈ati遍ons如果抗性園監(jiān)測(cè)確認(rèn)難了抗藥性承,矯正患措施可以例減緩害蟲(chóng)夾種群抗藥填性的傳播23Rem桂edi股al捎act拳ion夢(mèng)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(mèng)in敞af歌fec怪ted鑒ar沒(méi)easInc凡rea譽(yù)sed薦re常sis托tan租ce刺mon迅ito語(yǔ)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(cè)sta炸nce美國(guó)環(huán)保憲局措施包孕括:強(qiáng)制要石求對(duì)有稿懷疑的爐抗藥性鎮(zhèn)表現(xiàn)上纏報(bào)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(cè)在被影響臟區(qū)域終止Bt玉米銷翼售對(duì)抗藥性坑原因進(jìn)行奶調(diào)查24Com傭pli旗anc還ea付ssu來(lái)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(chóng)抗詞藥性管理球規(guī)則執(zhí)行烈耕種操作時(shí).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(píng)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(tīng)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(guó)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(duì)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(duì)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r(shí)esp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(duì)IRM策略的必涌要性,域怎樣完成捷要求,墊和種植者蜻對(duì)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(guó)環(huán)胞保局對(duì)棋鱗翅目席昆蟲(chóng)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(dòng)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(kāi)展閥規(guī)范性滔調(diào)查28Summ綱ary由ofE海PA’s時(shí)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(duì)鱗翅目位昆蟲(chóng)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(cè)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(duì)鱗翅目辮昆蟲(chóng)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(bào)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(cè)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(guó)環(huán)保狐局IRM項(xiàng)目的貧數(shù)據(jù)來(lái)抵源Pub勵(lì)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(shuō)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(bào)觸告專家(學(xué)泰術(shù)界,政們府,工業(yè)陷界)會(huì)議(顧哲問(wèn)團(tuán),研違討會(huì),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(guó)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(duì)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(wèn)to篇succ售ess席atE特PA美國(guó)環(huán)店保局(EPA丸)取得成功掏的關(guān)鍵Cond燦itio殺nal住(tim戲e-li喜mite遇d)r慘egis緩trat賓ions疏ena參ble鏈EPA紙toa煉dapt迎IRM滲req員uire洞ment宅sov榨ert蔬imeEPA到ne波got組iat率es蜘reg僑ula搏tor堡yc圖han失ges柄wi所th厘reg膠ist崗ran巧ts斜and輝gr璃owe羽ro哄rga話niz塞ati漸ons有時(shí)間限況制的注冊(cè)止可以使EPA適應(yīng)IRM隨著時(shí)息間變化階而產(chǎn)生綱的新要蠢求EPA經(jīng)常與閣注冊(cè)者粘,種植依者組織綢就規(guī)則句調(diào)整進(jìn)肺行協(xié)商33IRM因ch歪all餅eng巴es陪IRM面臨的臟挑戰(zhàn)Plan浮sar熄esp附ecif殘ict款oth賤e:Pest援/cro妥pCoun輛try/補(bǔ)regi嗓onBts違elec喘tion夾int喘ensi染tyw躺ill耗bea欣ffec逗ted勇by:Adop衰tion哈/dis邪trib悼utio姓nof濟(jì)aBtcro秋pCro敘ss-洪res握ist具anc伸ep滿ote嘩nti降al爹wit價(jià)hs夫imi禍lar斜pr銷