首页|异源韧皮部特异启动子在转基因枳中的表达

异源韧皮部特异启动子在转基因枳中的表达

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为了筛选在柑橘韧皮部中特异表达的启动子,构建GRP、Rolc、RSS1异源韧皮部特异启动子控制GUS报告基因的植物表达载体,转化枳[Poncirus trifoliata (L.) Raf.]上胚轴.GUS组织化学染色分析表明,3个启动子均显示了维管束组织特异性.其中,GRP启动子活性强,且具有严格的韧皮部组织特异性;Rolc启动子活性最强,在根和茎中呈组成型表达;RSS1启动子活性最弱.Real-time RT-PCR和GUS酶活性分析进一步证明了GRP启动子具有较强的韧皮部组织特异性,该启动子有望应用于柑橘抗黄龙病基因工程育种.
Expression Analysis of Three Phloem-specific Promoters in Transgenic Poncirus trifoliata
For screening phloem-specific promoters in citrus,the plant expression vectors containing GUS reporter cassette driven by Glycine-rich protein gene promoter(GRP)from Vigna unguiculata (Linn.)Walp (bean),Agrobacterium Rhizogenes RolC gene promoter (Rolc) from Agrobacterium rhizogenes and Sucrose synthase 1 gene (RSS1) promoter from Oryza sativa (rice) were constructed,and introduced into the Poncirus trifoliata (L.) Raf.seedling epicotyls by Agrobacterium-mediated transformation.Histochemical β-glucuronidase (GUS)analysis revealed vascular-specific expression of the GUS protein in citrus.The GRP promoter displayed strong and strict phloem-specific expression pattem in this study while the RSS 1 promoter could drive low levels of GUS gene expression in citrus.Strong constitutive expression of GUS driven by the Rolc promoter was detected in root and stem.These results were further confirmed by reverse transcription real-time polymerase chain reaction and quantitative GUS enzyme activity assays.The strong vascular-specific promoter i.e.GRP promoter provides a good candidate for targeting expression of antibacterial constructs to battle Citrus huanglongbing disease (HLB or citrus greening disease),associated with a phloem-limited Gram-negative bacterium

Poncirus trifoliataphloem-specificity promotergenetic transformationGUS

许兰珍、彭爱红、何永睿、姚利晓、雷天刚、刘小丰、姜国金、邹修平、陈善春

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西南大学柑橘研究所/中国农业科学院柑橘研究所,国家柑橘工程技术研究中心,国家柑橘品种改良中心,重庆400712

枳 韧皮部特异性启动子 遗传转化 GUS

国家自然科学基金长江学者和创新团队发展计划项目国家高科技研究发展计划(863计划)项目国家现代农业产业技术体系建设专项资金项目农业部公益性行业(农业)科研专项重庆市科委重点实验室专项经费项目重庆市自然科学基金中央高校基本科研业务费专项

31272150PCSIRT2011AA100205CARS-272010 03067-02-4CSTCCSTC2011JJA80029XDJK2011C003

2014

园艺学报
中国园艺学会 中国农业科学院蔬菜花卉研究所

园艺学报

CSTPCDCSCD北大核心
影响因子：1.127
ISSN：0513-353X
年,卷(期)：2014.41(1)
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