Maize transcription factor Zmdof1 involves in the regulation of Zm401 gene
Maize transcription factor Zmdof1 involves in the regulation of Zm401 gene
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玉米转录因子Zmdof1参与Zm401基因的调控
DOI：
10.1007/s10725-011-9651-5
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发表时间：
2012-04-01
影响因子：
4.2
通讯作者：
Yu, Jingjuan
中科院分区：
文献类型：
--
作者：
Chen, Xiyang;Wang, Dongxue;Yu, Jingjuan
Zmdof 1是玉米Dof转录因子家族基因的一员，参与PEPC基因的调控。Zm 401基因是从玉米中克隆的一个含有短开放阅读框（ORF）的基因，其启动子含有多个Zmdof 1识别位点（DOFCORE，AAAG）。Zm 401在玉米花药发育过程中具有重要作用，其中最长的ORF编码的蛋白Zm 401 p10定位于细胞核，是玉米花药发育所必需的。本研究克隆了Zmdof 1基因，表达谱分析表明Zmdof 1基因不仅参与玉米营养器官的发育，而且参与玉米花粉的成熟。Zmdof 1：：GFP融合蛋白在洋葱表皮细胞中的瞬时表达显示出核定位。Zm 401启动子的5′端缺失分析表明，-670到-510区域对启动子活性很重要。酵母单杂交系统中的反式激活试验证实Zmdof 1与Zm 401启动子中的AAAG元件具有强相互作用。PZm 401：：Gus构建体与UBI：：Zmdof 1的共转化导致GUS表达降低约40%。当Zmdof 1被转化时，由其天然启动子控制的Zm 401的异位表达导致的花粉生活力降低被恢复。定量RT-PCR分析表明，在PZm 401：：Zm 401/UBI：：Zmdof 1转基因烟草中，Zm 401的表达量显著降低，而Zmdof 1的表达量显著增加。结果表明，Zmdof 1在体外与Zm 401启动子相互作用，并在转基因烟草花粉中下调Zm 401。提出了花粉中Zmdof 1对Zm 401的可能调控机制。
Zmdof1 is a member of the maize Dof transcription factor family genes and participates in the regulation and control of the PEPC gene. The Zm401 gene, which contains short open reading frames (ORFs), has been cloned from maize, and its promoter contains several Zmdof1 recognition sites (DOFCORE, AAAG). Zm401 has an important role in anther development, and the protein encoded by the longest ORF, Zm401p10, localizes in the nucleus and is essential for maize anther development. In this study, we cloned Zmdof1, and expression pattern assay suggested that Zmdof1 has a role not only in nutrition organ development but also in maize pollen maturation. Transient expression of a Zmdof1::GFP fusion protein in onion epidermal cells showed a nuclear localization. 5′ deletion analysis of the Zm401 promoter showed that the region of −670 to −510 is important for promoter activity. Trans-activation assays in the yeast one-hybrid system confirmed that Zmdof1 had a strong interaction with AAAG elements in the Zm401 promoter. Co-transformation of a PZm401::Gus construct with UBI::Zmdof1 resulted in an approximately 40% decrease in GUS expression. Decrease of pollen viability resulting from ectopic expression of Zm401 controlled by its native promoter was recovered when Zmdof1 was transformed. Quantitative RT-PCR analysis showed that in PZm401::Zm401/UBI::Zmdof1 transgenic tobacco, the expression of Zm401 decreased significantly, coupled with an increase of Zmdof1 expression. The results indicated that Zmdof1 interacts with the Zm401 promoter in vitro and downregulates Zm401 in transgenic tobacco pollen. A probable regulatory mechanism of Zmdof1 to Zm401 in pollen was proposed.