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1、南方農(nóng)業(yè)學報JOURNALOFSOUTHERNAGRICULTURE2013,44(7):1081—1086ISSN2095—1191;CODENNNXAABhttp://www.nfnyxb.eliDOI:10.3969/j:issn.2095—1191.2013.7.1081農(nóng)桿菌介導的葡萄柚轉(zhuǎn)化再生體系建立研究韓美麗1,陸榮生1,林睿2,馬躍峰1,楊玉霞1,覃建林1(1廣西農(nóng)業(yè)科學院植保所/廣西作物病蟲害生物學重點實驗室,南寧530007;2廣西醫(yī)科大學,南寧530001)摘要:【目的】建立鄧肯葡萄柚上胚軸遺傳轉(zhuǎn)化再生體系,為葡萄柚基因轉(zhuǎn)化研究奠定基礎(chǔ)。【方法】以鄧肯
2、葡萄柚上胚軸為外植體,研究上胚軸不定芽和生根誘導、遺傳轉(zhuǎn)化的影響因子。【結(jié)果】在添=cJIITDZo.50~1.00mg/L的培養(yǎng)基上,上胚軸不定芽誘導率為72.7%~81.6%;無菌枝條的最適生根條件為1/2MT添加ABT6號1.0meCL,生根率可達81.3%。不同抗菌素中,以替門汀殺菌效果較好,且對外植體出芽無明顯抑制作用。轉(zhuǎn)化芽選擇初期,5~7d的暗培養(yǎng)有利于卡那霉素抗性芽的誘導發(fā)生。PcR檢測結(jié)果顯示,7株抗性植株中有6株呈陽性?!窘Y(jié)論】建立了鄧肯葡萄柚上胚軸植株再生繁殖體系,為其基因轉(zhuǎn)化奠定了基礎(chǔ)。關(guān)鍵詞:葡萄柚;不定芽;再生;轉(zhuǎn)化中圖分類號:$666.3文獻標
3、志碼:A文章編號:2095—1191(2013)07—1081—06EstablishmentregenerationsystemforCitrus.paradisiMacf.CVDuncanbyAgrobacterium-mediatedtransformationHANMei—lil,LURong—sheng1,LINRui2,MAYue-fen91,YANGYu-xia',QINJian-linl(1PlantProtectionResearchInstitute,GXAAS/GuangxiKeyLaboratoryofBiologyforCropDiseasesan
4、dInsectPests,Nanning530007,China;2GuangxiMedicalUniversity,Nanning530021,China)Abstract:【Objective】TransformationregenerationsystemofepicotylsderivedfromCitrusparadisiMacf.CVDuncanseedlingwereestablishedinordertoprovideusefulreferenceforitsgenetransformation.【Method】Epicotylsofseedlingwere
5、usedasexplantstoinvestigatetheinfluencefactorsoninductionanddifierentiationofadventitiousbuds.rootingofplantlets,andgenetictransformation.【ResuhlTheinductionrateofadventitiousbudsderivedfromepicotyleswererangedfrom72.7%to81.6%inmediawith0.50—1.00mg/LTDZ.Comparingthreetypesofgrowthregulator
6、s.thehigherrootingrate(81.3%)wereobservedin1/2MTmediumwith1.0mg/LofABT6rootingpowder.Amongdifferentantibiotics,ti—mentinshowedbettersterilizingeffectsthancefotaximesodiumandceftriaxonesodiumonagrobacterium—mediatedtrans—formationplantletsculture.Timentinhadnoinhibitiononbudsinductionofexpl
7、ants.Duringinitialtransformationstageofbuds,5—7daysofdarknessculturewasbeneficialtoinduceKananti-buds.Among7transgenicplantlets,6plantletsshowedpositiveinPCRdetection.【Conclusion]RegenerationsystemofCitrusparadisiMacf.evDuncanecopitylswase8一tablishedandprovide