Applied Plant and Soil Biology, Faculty of Biology, University of Vigo, Vigo, Spain.
Plant Physiol Biochem. 2011 Apr;49(4):413-9. doi: 10.1016/j.plaphy.2011.02.005. Epub 2011 Feb 19.
In the view of the economic importance of grapevine and the increasing threaten represented by vascular diseases, transgenic grapevine with enhanced tolerance could represent an attractive opportunity. Hitherto, constitutive promoters have been used generally to study the effects of transgene expression in grapevine. Given the fact that constitutive gene expression may be harmful to the host plant, affecting plant growth and development, the use of tissue -specific promoters restricting gene expression to tissues of interest and at given developmental stages could be more appropriate. For this purpose, we decided to study in grapevine the activity of the Eucalyptus gunnii CCR promoter that was previously reported to be vascular-preferential. We transformed grapevine with the "Sonication assisted Agrobacterium-mediated transformation" (SAAT) method and a construct where both GUS and GFP (green fluorescent protein) marker genes were under control of the EgCCR promoter. High GUS and GFP activities were found to be associated with the newly formed vascular tissues in stems, leaves and petioles of transformed grapevine, suggesting a preferential activity of the EgCCR promoter in the vascular tissues of grapevine. These results suggest the tissue-specificity of this promoter from eucalyptus is conserved in grapevine and that it could be used to drive expression of defense genes in order to enhance resistance against vascular pathogens.
鉴于葡萄在经济上的重要性,以及血管疾病带来的威胁日益增加,具有增强耐受性的转基因葡萄可能是一个有吸引力的机会。迄今为止,组成型启动子通常被用于研究转基因在葡萄中的表达效应。考虑到组成型基因表达可能对宿主植物有害,影响植物的生长和发育,使用组织特异性启动子将基因表达限制在感兴趣的组织和特定的发育阶段可能更为合适。为此,我们决定研究先前报道在血管组织中优先表达的桉树 CCR 启动子在葡萄中的活性。我们使用“超声辅助农杆菌介导转化”(SAAT)方法和一个构建体转化葡萄,该构建体中的 GUS 和 GFP(绿色荧光蛋白)标记基因均受 EgCCR 启动子的控制。在转化葡萄的茎、叶和叶柄中新形成的维管束组织中发现了高的 GUS 和 GFP 活性,这表明 EgCCR 启动子在葡萄的维管束组织中具有优先活性。这些结果表明,来自桉树的这种启动子在葡萄中的组织特异性是保守的,它可以用于驱动防御基因的表达,以增强对血管病原体的抗性。