基因枪转化小麦幼胚的再生培养与转基因植株的获得

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1、西北植物学报2003Κ23;9ΓΠ1565—1570ActaBot.Boreal.-Occident.Sin.文章编号Π100024025;2003Γ0921565206Ξ基因枪转化小麦幼胚的再生培养与转基因植株的获得尹钧Κ任江萍Κ宋丽Κ李磊Κ师学珍Κ郭向云;河南农业大学国家小麦工程技术研究中心Κ郑州450002Γ摘要Π以小麦幼胚为受体Κ用基因枪法对Trx2S反义基因;目的基因Γ和Bar基因;标记基因Γ进行了共转化Κ以轰击后的小麦幼胚为实验材料Κ对幼胚培养的基本培养基、分化和生根培养基进行了筛选优化Λ结果表明Π4种基本培养基中ΚL3培养基的成愈率最高Κ且增殖速度快

2、ΜMS培养基次之Λ以L3为基本培养基Κ分化培养基中添加NAA1mg·L-1和ZT2mg·L-1配比对愈伤组织诱导分化的效果最好Κ分化率达到50%以上Λ1ƒ2MS培养基中添加IAA0.8mg·L-1的生根效果好Κ且移栽成活率高Λ以优化的培养方案对来自7个小麦品种的幼胚进行转化与再生培养Κ多数品种的出愈率都达到90%以上Κ分化率在40%以上Κ并在5个品种上获得再生植株Κ经检测证实在4个品种上获得转基因再生植株Λ关键词Π小麦幼胚Μ基因枪转化Μ再生植株培养Μ转基因再生植株中图分类号ΠQ943.1文献标识码ΠARegenerationculturefrombombarded

3、immatureembryosofwheatsandtransgenicplantYINJunΚRENJiang2pingΚSONGLiΚLILeiΚSHIXue2zhenΚGUOXiang2yun;NationalEngineeringResearchforWheatΚHenanAgriculturalUniversityΚZhengzhou450002ΚChinaΓAbstractΠAntisenseTrx2SgeneandBargentwereco2transformedtoimmatureembryosofwheatviami2croprojectile2

4、bombardment.ThemediaforinducingcallusΚplantregenerationfrombombardedimmatureembryoswerestudiedforhighlyefficientplantregeneration.ThebasicmediainexperimentswereincludingMSΚL3ΚN6B5ΚMSandB5.L3isthemostidealmediumforinducingcallusanddifferentiationinthe4usedmediawithainducingcallusfreque

5、ncymorethan90%.ThesecondisMSmediumΚitsfrequency80%～90%.TheoptimumhormoneconcentrationaddedtoL3mediumforcallusdifferentiationwasNAA;naph2thaleneaceticacidΓ1mg·L-1ZT;zeatinΓ2mg·L-1withacallusdifferentiationfrequencymorethan50%.IAA;indole232aceticacidΓ0.8mg·L-1addedtotherootingmedium;1ƒ2

6、MSΓwasoptimumfortherootingofbudsandforefficienttransplantation.Theimmatureembryosfrom7genotypeswerebombardedandculturedontheoptimummedia.Thefrequencyofcallusinductionanddifferentiationreachedtomorethan90%and40%inmostgenotypesrespectively.Theregeneratedplantsfromtheimmatureembryosof5ge

7、notypesandtransgenicplantsfrom4genotypeswereobtainedbyPCRandPCR2Southerntest.KeywordsΠimmatureembryosofwheat;TriticumaestivumL.ΓΜmicroprojectile2bombardmentΜregenerat2edplantcultureΜtransgenicplants收稿日期Π2002212225Μ修改稿收到日期Π2003204214基金项目Π国家转基因植物研究与产业化专项;J20002B2019Γ和河南省杰出人才创新基金;1210007

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