• ISSN 1008-505X
  • CN 11-3996/S

水稻不同器官寡肽运输基因家族启动子活性分析

Analysis of promoter activity of rice oligopeptide transport gene family in different organs

  • 摘要:
    目的 分析寡肽运输基因在水稻不同器官中的表达特异性,以加深对水稻营养物质的吸收、运输与分配调节的了解。
    方法 以粳稻日本晴基因组为模板,分别克隆水稻寡肽运输基因家族PT亚家族9个基因的启动子序列,并将其插入到pCAMBIA1301载体的葡糖醛酸酶(GUS)蛋白编码区前,形成启动子诱导GUS蛋白表达载体,通过农杆菌,制备9个水稻寡肽运输基因启动子分别诱导GUS蛋白表达的转基因水稻,在自然生长季进行田间试验,于抽穗开花期,分别剪取水稻根、茎、叶、小穗等器官样品,浸泡在GUS染色液中进行组织化学染色,体视镜观察、拍照。分析水稻基因表达数据库RiceXPro中9个水稻寡肽运输基因在不同器官的表达谱,采用PlantCare数据库对9个水稻寡肽运输基因的启动子序列的顺式作用元件进行分析。
    结果 9个水稻寡肽运输基因家族启动子均可以诱导GUS蛋白在水稻节的部位强烈表达,其中7个可以诱导GUS蛋白在水稻内稃和外稃表达,分别是OsOPT1OsOPT2OsOPT4OsOPT5OsOPT7OsOPT8OsOPT9。RiceXPro水稻基因表达数据库中的表达谱显示OsOPT5OsOPT6的表达具有很强的特异性,这两个基因都是只在花药强烈表达,在其它器官中未检测到表达。另外,在水稻寡肽运输基因家族启动子中发现多个赤霉素和脱落酸响应元件。
    结论 水稻寡肽运输基因家族基因的启动子活性具有强烈的器官特异性,9个水稻寡肽运输基因的启动子均在水稻茎节的部位强烈诱导GUS蛋白表达,说明这些基因参与了水稻不同器官间营养物质的交换和分配,并且在赤霉素和脱落酸信号响应中发挥作用。

     

    Abstract:
    Objectives We analyzed the spatiotemporal expression of rice oligopeptide transport genes in different organs of rice, in order to understand the mechanism of the absorption, transport and distribution of oligopeptides and other nutrients in rice.
    Methods Employing the genome of japonica rice variety Nipponbare as the template, the promoter sequences of the nine genes belonging to the PT subfamily of the rice oligopeptide transport gene family were separately cloned and inserted in front of the glucuronidase (GUS) protein coding region in the pCAMBIA1301 vector, thereby forming the promoter-induced GUS protein expression vectors. Through Agrobacterium, transgenic rice plants with GUS protein expression respectively induced by the nine rice oligopeptide transport gene promoters were prepared. Field experiment was carried out during the natural growing season. At the heading and flowering stage, organ samples such as roots, stems, leaves, and spikelets of rice were cut and immersed in GUS staining solution for histochemical staining. Stereomicroscopic observation and photography were conducted. The expression profiles of the nine rice oligopeptide transport genes in different organs in the rice gene expression database RiceXPro were analyzed, and the cis-acting elements of the promoter sequences of the nine rice oligopeptide transport genes were analyzed using the PlantCare database.
    Results The nine promoters of rice oligopeptide transport gene family could induce GUS protein to express strongly in rice nodules, and seven of them could induce GUS protein to express strongly in rice inner and outer husks, which were OsOPT1, OsOPT2, OsOPT4, OsOPT5, OsOPT7, OsOPT8 and OsOPT9. The expression profile of RiceXPro rice gene expression database showed that the expression of OsOPT5 and OsOPT6 had strong specificity, which were strongly expressed only in the late stage of anther development and were not detected in other organs. In addition, multiple GA and abscisic acid response elements were found in the promoters of rice oligopeptide transport gene family.
    Conclusions The promoter activity of the rice oligopeptide transporter gene family has strong organ-specificity. The expression of GUS protein is strongly induced in the node region of rice, where 9 rice oligopeptide transporter gene promoters are located, indicating that these genes participate in the exchange and allocation of nutrients among different organs of rice and play a role in the response to auxin and abscisic acid signals.

     

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