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园艺学报 ›› 2008, Vol. 35 ›› Issue (4): 487-492.

• 果树 • 上一篇    下一篇

梨韧皮部特异表达启动子AtSUC2驱动下的GUS基因的转化和表达

孙清荣1*;孙洪雁1;赵 衍2;海门得·茹斯2;戴维斯·罗伯特2   

  1. 1 山东省果树研究所,山东泰安271000;2 美国农业部百思维尔农业研究中心,植物分子病理实验室,马里兰20705,美国)
  • 收稿日期:2007-12-27 修回日期:1900-01-01 出版日期:2008-04-25 发布日期:2008-04-25
  • 通讯作者: 孙清荣

Transgene Expression in Pear (Pyrus communis L.) Driven by Phloem-specific Promoter AtSUC2

SUN Qing-rong1* , SUN Hong-yan1 , ZHAO Yan2, HAMMOND Rosemarie2, and DAVIS Robert E.2   

  1. (1Shandong Institute of Pomology, Tai'an, Shandong 271000, China; 2Molecular Plant Pathology Laboratory, USDA-Agriculture Research Service, Beltsville, MD 20705, U.S.A.)
  • Received:2007-12-27 Revised:1900-01-01 Online:2008-04-25 Published:2008-04-25
  • Contact: SUN Qing-rong

摘要:

以 ‘Old Home’ 梨无菌苗叶片为外植体, 利用根癌农杆菌介导将专一在韧皮部表达启动子AtSUC2 驱动下的GUS 基因转入 ‘Old Home’中。筛选出了‘Old Home’ 梨叶片高效遗传转化的最佳条件:农杆菌和外植体在液体培养基上共培比在固体培养基上共培转化率提高5倍。 PCR分析和Southern 杂交鉴定表明GUS 基因已整合到转基因植株的基因组中,组织化学检测和Western 杂交鉴定证明了GUS 基因在转基因植株韧皮部中的表达。

关键词: 梨, 启动子, 韧皮部特异表达, 基因转化

Abstract:

Leaves of 'Old Home' Pear (Pyrus communis L.) in vitro were selected as explants. GUS (β-glucuronidase) reporter gene (uidA) under the control of the Arabidopsis sucrose-H+ symporter gene (AtSUC2) promoter which is phloem-specific was introduced into 'Old Home' by Agrobacterium tumefaciens. Optimal conditions were established for efficient transformation from leaf explants of a pear cultivar, 'Old Home'. High transformation efficiency was achieved by an improved induction stage following initial Agrobacterium infection. In the induction stage, Agrobacterium cells and parent leaf explants were co-cultivated on a liquid induction medium, which yielded a five-fold increase of transformation frequency over conventional co-cultivation on a solid medium. The integration was checked by PCR and Southern hybridization. The expression of uidA gene was analyzed by GUS histochemical detection and Western blot analysis.

Key words: pear, promoter, phloem-specific expression, transgene

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