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园艺学报 ›› 2014, Vol. 41 ›› Issue (1): 1-8.

• 果树 •    下一篇

异源韧皮部特异启动子在转基因枳中的表达

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*   

  1. 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
  • 出版日期:2014-01-25 发布日期:2014-01-25
  • 基金资助:

    国家自然科学基金项目(31272150);长江学者和创新团队发展计划项目(PCSIRT);国家高科技研究发展计划(863 计划)
    项目(2011AA100205);国家现代农业产业技术体系建设专项资金项目(CARS-27);农业部公益性行业(农业)科研专项(2010 03067-02-4);
    重庆市科委重点实验室专项经费项目(CSTC);重庆市自然科学基金项目(CSTC2011JJA80029);中央高校基本科研业务费专项
    (XDJK2011C003)

Expression Analysis of Three Phloem-specific Promoters in Transgenic Poncirus trifoliata

XU Lan-zhen,PENG Ai-hong,HE Yong-rui,YAO Li-xiao,LEI Tian-gang,LIU Xiao-feng,JIANG Guo-jin,ZOU Xiu-ping*,and CHEN Shan-chun*   

  1. Citrus Research Institute,Southwest University;Citrus Research Institute,Chinese Academy of Agricultural Sciences;National Citrus Engineering Research Center;National Center for Citrus Varieties Improvement,Chongqing 400712,China
  • Online:2014-01-25 Published:2014-01-25

摘要: 为了筛选在柑橘韧皮部中特异表达的启动子,构建GRP、Rolc、RSSl 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;RSSl 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。

关键词: 枳, 韧皮部特异性启动子, 遗传转化, GUS

Abstract: 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(RSSl)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 pattern in this study while
the RSS1 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

Key words: Poncirus trifoliata, phloem-specificity promoter, genetic transformation, GUS

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