轉(zhuǎn)基因食品安全：挑戰(zhàn)與機(jī)遇

轉(zhuǎn)基因食品安全：挑戰(zhàn)與機(jī)遇SafetyIssuesofGeneticallyModifiedFoods:ChallengesandOpportunities農(nóng)業(yè)部轉(zhuǎn)基因生物產(chǎn)品成分監(jiān)督檢驗(yàn)測(cè)試中心（上海）（農(nóng)業(yè)部認(rèn)可和國(guó)家計(jì)量認(rèn)證）SupervisionandTestingCenterforGeneticallyModifiedOrganisms,MinistryofAgriculture(AccreditedbyMOAandtheNMA)上海交通大學(xué)轉(zhuǎn)基因生物分子特征驗(yàn)證測(cè)試中心（CNAS認(rèn)可）CenterfortheMolecularCharacterizationofGeneticallyModifiedOrganisms,ShanghaiJiaoTongUniversity（AccreditedbyCNAS）2.轉(zhuǎn)基因食品檢測(cè)現(xiàn)狀和面臨的挑戰(zhàn)ChallengesofGMFoodDetection轉(zhuǎn)基因食品及其安全GMFoodsandTheirSafety3.我們目前的研究工作StudiesinOurLaboratory報(bào)告內(nèi)容/OutlineIntroductionofGMODetection轉(zhuǎn)基因食品及其安全GMFoodsandSafety轉(zhuǎn)基因作物應(yīng)用迅猛發(fā)展StatusofGMCropPlantingWorldwide國(guó)際農(nóng)業(yè)生物技術(shù)應(yīng)用服務(wù)組織(ISAAA)中國(guó)已成為世界最主要的轉(zhuǎn)基因植物研發(fā)國(guó)Chinahasbecomeoneoftheworld'sleadersinR&DofGMplantsa.Chinalaunchedspecialbreedingprojectongeneticallymodifiedorganismsin2008a.2008年，我國(guó)開始實(shí)施“轉(zhuǎn)基因生物新品種培育”重大專項(xiàng)c.AlargenumberofdomesticallydevelopedGMcropsareexpectedtobereleasedforthetestinthefieldandenvironmentinthenext10yearsc.預(yù)計(jì)在未來(lái)10年內(nèi)我國(guó)將會(huì)有大量轉(zhuǎn)基因作物進(jìn)入田間實(shí)驗(yàn)和環(huán)境釋放等階段b.ChinaissuedforthefirsttimethecertificatestotwoGMriceandoneGMmaizelinesin2009.Allcertificateswereapprovedagainin2014b.2009年首次頒布了轉(zhuǎn)基因抗蟲水稻和轉(zhuǎn)植酸酶基因玉米的安全證書。2014年它們獲得續(xù)申請(qǐng)中國(guó)轉(zhuǎn)基因作物的種植現(xiàn)狀GMCropPlantinginChina轉(zhuǎn)基因作物種類（GMCrops）種植面積（公頃）Area(hectare)性狀（Traits）棉花（Cotton）3900000抗蟲（Insectresistant）木瓜（Papaya）8500抗病毒（Virusresistant）楊樹（Poplar）543抗蟲（Insectresistant）CliveJames,2014國(guó)際農(nóng)業(yè)生物技術(shù)應(yīng)用服務(wù)組織(ISAAA)8我國(guó)安全證書發(fā)放情況

CertificatesIssuedinChina截止目前，共批準(zhǔn)發(fā)放7種轉(zhuǎn)基因作物安全證書a.1997年耐儲(chǔ)存番茄（證書失效）b.1997年抗蟲棉花（證書有效，商業(yè)化種植）c.1999年改變花色矮牽牛（證書失效）d.1999年抗病辣椒（證書失效）e.2006年抗病番木瓜（證書有效，商業(yè)化種植）f.2009年抗蟲水稻和轉(zhuǎn)植酸酶玉米（證書有效，未商業(yè)化種植）Sofar,7certificateshavebeenissueda.1997,longshelflifetomato（expired）b.1997，Btcotton（valid,commercialized）c.1999,pigmentedpetunia（expired）d.1999,diseaseresistantpepper（expired）e.2006,virusresistantpapaya（valid,commercialized）f.2009,Btriceandphytaseoverexpressingmaize(valid,notcommercializedyet）中國(guó)進(jìn)口轉(zhuǎn)基因作物的種類及事件TypesandEventsofGMCropImportedinChinaUSDA-FASGAINReportNumber:14032.China-PeoplesRepublicofAgriculturalBiotechnologyAnnual美國(guó)農(nóng)業(yè)部外國(guó)農(nóng)業(yè)局2015年1月5日發(fā)布的《中國(guó)農(nóng)業(yè)生物技術(shù)年報(bào)》5種作物41個(gè)轉(zhuǎn)化事件5crops41events中國(guó)進(jìn)口轉(zhuǎn)基因作物的性狀TraitsofGMCropImportedinChinaUSDA-FASGAINReportNumber:14032.China-PeoplesRepublicofAgriculturalBiotechnologyAnnual美國(guó)農(nóng)業(yè)部外國(guó)農(nóng)業(yè)局2015年1月5日發(fā)布的《中國(guó)農(nóng)業(yè)生物技術(shù)年報(bào)》中國(guó)進(jìn)口轉(zhuǎn)基因作物的研發(fā)者DevelopersofGMCropsImportinChinaUSDA-FASGAINReportNumber:14032.China-PeoplesRepublicofAgriculturalBiotechnologyAnnual美國(guó)農(nóng)業(yè)部外國(guó)農(nóng)業(yè)局2015年1月5日發(fā)布的《中國(guó)農(nóng)業(yè)生物技術(shù)年報(bào)》中國(guó)轉(zhuǎn)基因玉米的進(jìn)口數(shù)量AmountsofImportedGMmaizeinChina2011201220132014World全球1.755.213.261.99美國(guó)USA1.695.112.971.01烏克蘭Ukraine000.110.57泰國(guó)Thailand00.0200.29百萬(wàn)噸/MillionTonUSDA-FASGAINReportNumber:14032.China-PeoplesRepublicofAgriculturalBiotechnologyAnnual美國(guó)農(nóng)業(yè)部外國(guó)農(nóng)業(yè)局2015年1月5日發(fā)布的《中國(guó)農(nóng)業(yè)生物技術(shù)年報(bào)》中國(guó)轉(zhuǎn)基因大豆的進(jìn)口數(shù)量AmountsofImportedGMSoybeaninChina2011201220132014World全球52.6358.3863.4062.87巴西Brazil20.6223.8931.8131.83美國(guó)USA22.3525.9722.2622.25阿根廷Argentina7.845.906.125.82百萬(wàn)噸/MillionTonUSDA-FASGAINReportNumber:14032.China-PeoplesRepublicofAgriculturalBiotechnologyAnnual美國(guó)農(nóng)業(yè)部外國(guó)農(nóng)業(yè)局2015年1月5日發(fā)布的《中國(guó)農(nóng)業(yè)生物技術(shù)年報(bào)》中國(guó)轉(zhuǎn)基因棉花的進(jìn)口數(shù)量AmountsofImportedGMCottonsinChina2011201220132014World全球3.375.144.152.18印度India1.011.441.190.74美國(guó)USA0.981.461.150.50澳大利亞Australia0.530.820.800.48噸/TonUSDA-FASGAINReportNumber:14032.China-PeoplesRepublicofAgriculturalBiotechnologyAnnual美國(guó)農(nóng)業(yè)部外國(guó)農(nóng)業(yè)局2015年1月5日發(fā)布的《中國(guó)農(nóng)業(yè)生物技術(shù)年報(bào)》中國(guó)轉(zhuǎn)基因棉花的出口數(shù)量AmountsofGMCropExportedfromChina2011201220132014World全球2569817558673313121孟加拉國(guó)Bangladesh3678504104391越南Vietnam460079025062383朝鮮NorthKorea5631320520821958噸/TonUSDA-FASGAINReportNumber:14032.China-PeoplesRepublicofAgriculturalBiotechnologyAnnual美國(guó)農(nóng)業(yè)部外國(guó)農(nóng)業(yè)局2015年1月5日發(fā)布的《中國(guó)農(nóng)業(yè)生物技術(shù)年報(bào)》轉(zhuǎn)基因生物GMO

增產(chǎn)Increasedagriculturalproductivity減少殺菌劑用量Reducedpesticideuse脫貧溫飽Reducedpovertyandhunger

保護(hù)生態(tài)Protectedbiologicaldiversity減少CO2排放ReducedtheemissionsofCO2社會(huì)和環(huán)境效應(yīng)SocialandEnvironmentalBenefits轉(zhuǎn)基因生物GMO政治Political宗教Religious技術(shù)Technical食品安全Foodsafety環(huán)境安全Environ-mentsafetyEconomic,socialandculturalcontext顧慮和擔(dān)憂ConcernsandWorries爭(zhēng)論的焦點(diǎn)和實(shí)質(zhì)FocusandEssenceoftheDebate科學(xué)認(rèn)知Scientificcognition技術(shù)認(rèn)同Technicalapproval其他Others安全否Safeornot?風(fēng)險(xiǎn)分析

RiskAnalysis風(fēng)險(xiǎn)評(píng)估RiskAssessment風(fēng)險(xiǎn)交流RiskManagement風(fēng)險(xiǎn)管理RiskCommunication風(fēng)險(xiǎn)評(píng)估

RiskAssessment分子特征食用安全環(huán)境安全Labeling轉(zhuǎn)基因食品的安全性

GMFoodSafetya.FDAWHO,andFAOclaimthatthenutritionandsafetyofGMfoodsaresubstantialequivalencetotraditionalfoodsaslongastheyareofficiallyapproveda.美國(guó)食品與藥品管理局（FDA），世界衛(wèi)生組織和世界糧農(nóng)組織（WHO/FAO）認(rèn)為：經(jīng)過(guò)審批的轉(zhuǎn)基因食品，營(yíng)養(yǎng)和安全上跟傳統(tǒng)食品“實(shí)質(zhì)等同”b.Sofar,thereisnoevenonefoodsafetyaccidentcaseresultfromconsumingGMfoodsb.到目前為止，沒(méi)有一例食品安全事故是由轉(zhuǎn)基因食品造成的c.LabelingGMisonlyamodeofregulation,whichhasnothingtodowithfoodsafetyitselfc.轉(zhuǎn)基因標(biāo)記是一種管理模式，與其食品安全本身無(wú)關(guān)CurrentStatusofGMODetectionCurrentChallengesofGMODetection2.轉(zhuǎn)基因食品檢測(cè)現(xiàn)狀和面臨的挑戰(zhàn)ChallengesofGMFood

Detection基于蛋白質(zhì)的檢測(cè)Proteindetection酶聯(lián)免疫吸附法ELISA免疫試紙條Lateralflowstrip轉(zhuǎn)基因食品的檢測(cè)策略StrategyofGMODetection核苷酸提取DNAExtraction定性聚合酶鏈?zhǔn)椒磻?yīng)QualitativePCR不含轉(zhuǎn)基因成分NoGMcontent陰性Negative陽(yáng)性Positive含轉(zhuǎn)基因成分ContainGMcontent轉(zhuǎn)基因成分含量GMOcontents（%）

QuantitativePCR基于核苷酸的檢測(cè)Nucleicaciddetection定量PCR基于蛋白質(zhì)的檢測(cè)方法的缺陷DisadvantagesofProtein-basedGMDetectionMethodsa.Onlyidentifythepresenceofthetransgenicproteinbutdoesnotdifferentiatebetweendifferenteventscontainingthesametransgenicconstruct.a.只能鑒定轉(zhuǎn)基因蛋白的存在,但不能區(qū)分含有相同蛋白的不同轉(zhuǎn)基因事件b.Someproteinsareinstableandmostofthemarenearlyimpossibletobereliablydetectedinprocessedfoods.b.一些蛋白質(zhì)在加工過(guò)程中易降解，因而不可能被檢測(cè)到基于DNA檢測(cè)方法的缺陷DisadvantagesofDNA-basedGMDetectionMethodsa.設(shè)計(jì)引物時(shí)需要預(yù)先獲得轉(zhuǎn)化事件的DNA序列

b.相對(duì)定量，需要標(biāo)準(zhǔn)曲線

c.需要有證標(biāo)準(zhǔn)物質(zhì)進(jìn)行定量a.PriorinformationontheeventisneededtodesignspecificPCRprimers.b.Relativequantitationandstandardcurveisneeded.

c.Certificatedreferencematerialsareneeded.基于DNA的檢測(cè)方法更可靠，應(yīng)用更廣泛DNA-baseddetectionismorereliableandwidelyused轉(zhuǎn)基因食品檢測(cè)面臨的挑戰(zhàn)ChallengesofGMFoodDetectionb.Un-authorizedGMO:broadscreeningisessentiala.StackedGMO:singledetectionisnotenoughc.NewTechnologyGeneratedGMO:newdetectionmethodsneededb.非法轉(zhuǎn)基因事件廣泛的篩查必不可少a.復(fù)合轉(zhuǎn)基因事件用檢測(cè)單一事件方法無(wú)效c.新技術(shù)產(chǎn)生的轉(zhuǎn)基因事件傳統(tǒng)的檢測(cè)技術(shù)無(wú)法檢測(cè)復(fù)合轉(zhuǎn)基因事件越來(lái)越多StackedGMCropEventsAreIncreasingCliveJames,2014國(guó)際農(nóng)業(yè)生物技術(shù)應(yīng)用服務(wù)組織(ISAAA)復(fù)合轉(zhuǎn)基因作物的由來(lái)WaystoObtainStackedGMCrops雜交共轉(zhuǎn)化同時(shí)轉(zhuǎn)化多個(gè)性狀再轉(zhuǎn)化USDA-FASGAINReportNumber:14032.China-PeoplesRepublicofAgriculturalBiotechnologyAnnual美國(guó)農(nóng)業(yè)部我國(guó)農(nóng)業(yè)局2015年1月5日發(fā)布的《中國(guó)農(nóng)業(yè)生物技術(shù)年報(bào)》a.ProductofMonsantoandDowAgroSciencesb.Contains6insectresistanceand2herbicidetolerancegenesc.ImportapprovedbyKoreaandTaiwan.a.孟山都和陶氏產(chǎn)品b.含6

個(gè)抗蟲基因和2個(gè)抗除草劑基因c.獲得韓國(guó)和臺(tái)灣的進(jìn)口許可一個(gè)著名的復(fù)合轉(zhuǎn)基因玉米ASampleofStackedGMMaizeUn-authorizedGMCrops

GMplantsthatarenotauthorizedforuseasfoodorfeedinatleastonecountryHolst-Jensenetal.,BiotechnolAdv30(2012)1318–1335非法轉(zhuǎn)基因事件沒(méi)有獲得一個(gè)國(guó)家批準(zhǔn)用于食品和飼料的轉(zhuǎn)基因事件來(lái)源a.Asynchronousauthorizations:riceLL601(USAandJapan)a.不對(duì)稱的批準(zhǔn)：LL601轉(zhuǎn)基因水稻c.Failureinthequalitycontrolattheseedproductionlevel:Bt10maizec.種子制備過(guò)程的質(zhì)量控制失?。築t10玉米b.Escapefromfield-trialreleasesb.田間試驗(yàn)不正當(dāng)流失非法轉(zhuǎn)基因事件Un-authorizedGMCropsBroedersetal(2012)JBiomedBiotechdoi:10.1155/2012/402418NewTechnologyDerivedGMCrops:GMplantsgeneratedfromothernewbiotechnologyratherthanconventionalDNAinsertionConventionalDNAinsertioncassette:consistsofaregulatorypromoterregion,acodingsequence(trait),andaterminator新技術(shù)產(chǎn)生的轉(zhuǎn)基因事件NewTechnologyDerivedGMCropsOligonucleotidedirectedmutagenesisRNA-dependentDNAmethylationzincfingernucleaseLusseretal.NatureBiotechnol,2012,30(3):231-239.b.這些技術(shù)的產(chǎn)品用傳統(tǒng)的轉(zhuǎn)基因檢測(cè)方法無(wú)法檢測(cè)TheproductsofthesearenotdetectableoridentifiablewithstandardGMOdetectionmethodsa.新的技術(shù)包括鋅指酶、TALENT、CRISPR等基因編輯技術(shù)。EmergingnewtechnologiesincludegeneeditingonessuchasZFN,TALENT,andCRISPRdetectionmethodsHowtoDealwithTheseChallenges如何面對(duì)這些挑戰(zhàn)a.Changetheone-by-oneapproachpresentlyusedinGMOdetectionandidentification,itwillbecomepracticallyimpossibleandtoocostly.a.改變一對(duì)一的檢測(cè)模式，研發(fā)多重復(fù)合轉(zhuǎn)基因事件的檢測(cè)技術(shù)b.Thereisanurgentneedforthedevelopmentofaccurate,reliableandcosteffectivedetectionmethodsforbothscreeningandquantificationofalltheseGMOs.b.迫切需要研發(fā)準(zhǔn)確、可靠和經(jīng)濟(jì)有效的定性篩選檢測(cè)和定量檢測(cè)技術(shù)方法Future

ProspectsStudiesfromOurLaboratory3.我們目前的研究工作StudiesinOurLaboratoryResearchInterests研究方向a.MolecularcharacterizationofGMOsBasedontheprinciplesofsystembiologya.轉(zhuǎn)基因生物分子特征的識(shí)別基于系統(tǒng)生物學(xué)原理的識(shí)別b.DevelopmentofhighthroughputqualitativeandquantitativemethodsforGMOdetectionAccurate,reliableandeconomicscreeningmethodAbsolutequantificationb.研發(fā)轉(zhuǎn)基因生物定性和定量的高通量檢測(cè)方法準(zhǔn)確、可靠和經(jīng)濟(jì)有效的高通量篩選檢測(cè)絕對(duì)定量方法基因組水平分子特征識(shí)別Genome-wideMolecularCharacterizationYangetal.SciRep.2013.No.2839TT51-1T1c-19ModelforNGSDataAnalysis代謝組水平分子特征識(shí)別Metabolome-wideMolecularCharacterizationHuet.al.ScientificReports2014.RiceseedmetabolomeLinet.al.JournalofIntegrativePlantBiology2014Soybeanseedmetabolome代謝組水平分子特征識(shí)別Metabolome-wideMolecularCharacterizationRaoet.al.Metabolomics.2014MaizeseedmetabolomeRaoetal.Unpublished微滴PCR毛細(xì)管電泳技術(shù)MPIC

（MicrodropletPCRImplementedCapillarygelelectrophoresis）a.同時(shí)檢測(cè)14個(gè)轉(zhuǎn)基因事件的24個(gè)靶標(biāo)基因Simultaneouslydetect24GMtargetsfrom14GMeventswitharelativeLODof0.1%（w/w）Guoetal.AnalChem,2011,83:1579-1586四重復(fù)合簡(jiǎn)并PCR技術(shù)PCRApproacha.定靶于8個(gè)性狀基因的9個(gè)DNA序列b.同時(shí)檢測(cè)到超過(guò)90個(gè)的合法轉(zhuǎn)基因作物a.FourPCRsystemstargetingonnineDNAsequencesfromeighttraitgenesb.Simultaneouslydetectmorethan90approvedGMOevents.c.LODis80copyGuoetal.,FoodChem.2011.132:1566–1573Shaoetal.AnalChem,2014,86(2):1269-1276芯片篩選檢測(cè)技術(shù)MACRO：multiplexamplificationonachipwithreadoutonanoligomicroarraya.芯片上含有91個(gè)靶標(biāo)轉(zhuǎn)基因作物成分的DNA序列，可以涵蓋目前97.1%商業(yè)化的轉(zhuǎn)基因事件a.Contains91targets(18universalelements,20exogenousgenes,45events,and8endogenousreferencegenes)thatcovers97.1%ofcommercializedGMcrops.SampleDilutionPartitionPCRReadout數(shù)字PCRDigitalPCRforAbsoluteGMOQuantification數(shù)字PCR平臺(tái)DigitalPCRPlatform有證標(biāo)準(zhǔn)物質(zhì)的定值轉(zhuǎn)基因作物拷貝數(shù)分析CopyNumberAnalysisofExogenousGeneinGMOsJiangetal.UnpublishedData轉(zhuǎn)基因產(chǎn)品的絕對(duì)定量QuantificationofGMProductsJiangetal.UnpublishedDataRMmethodtestedvalue(%)bias(%)ddPCRcdPCR3D-dPCRddPCRcdP