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Identification of the Regulatory Region Responsible for Vascular Tissue–Specific Expression in the Rice Hd3a Promoter

机译:水稻Hd3a启动子中负责血管组织特异性表达的调控区的鉴定

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摘要

Flowering time is determined by florigens. These genes include, Heading date 3a (Hd3a) and Rice FT 1 (RFT1) in rice, which are specifically expressed in the vascular tissues of leaves at the floral transition stage. To study the cis-regulatory elements present in the promoter region of Hd3a, we generated transgenic plants carrying the 1.75-kb promoter fragment of Hd3a that was fused to the β-glucuronidase (GUS) reporter gene. Plants expressing this construct conferred a vascular cell-specific expression pattern for the reporter gene. However, GUS was expressed in leaves at all developmental stages, including the early seedling stage when Hd3a was not detected. Furthermore, the reporter was expressed in roots at all stages. This suggests that the 1.75-kb region lackings cis-elements that regulate leaf-specific expression at the appropriate developmental stages. Deletion analyses of the promoter region indicated that regulatory elements determining vascular cell-specific expression are present in the 200-bp region between -245 bp and -45 bp from the transcription initiation site. By transforming the Hd3a–GUS construct to rice cultivar ‘Taichung 65’ which is defective in Ehd1, we observed that Ehd1 is the major regulatory element that controls Hd3a promoter activity.
机译:开花时间由花序决定。这些基因包括水稻中的抽穗期3a(Hd3a)和水稻FT 1(RFT1),它们在花期过渡期在叶片的维管组织中特异性表达。为了研究Hd3a启动子区域中的顺式调控元件,我们生成了带有Hd3a的1.75-kb启动子片段的转基因植物,该片段与β-葡萄糖醛酸酶(GUS)报告基因融合。表达该构建体的植物为报告基因赋予了血管细胞特异性表达模式。但是,GUS在所有发育阶段都在叶片中表达,包括未检测到Hd3a的幼苗早期。此外,记者在各个阶段都扎根。这表明1.75kb区域缺少在适当的发育阶段调节叶片特异性表达的顺式元件。启动子区域的缺失分析表明,决定血管细胞特异性表达的调控元件存在于距转录起始位点-245 bp和-45 bp之间的200 bp区域中。通过将Hd3a–GUS构建体转化为Ehd1有缺陷的水稻品种“ Taichung 65”,我们观察到Ehd1是控制Hd3a启动子活性的主要调控元件。

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