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Maize transcription factor Zmdof1 involves in the regulation of Zm401 gene

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Abstract

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.

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Acknowledgments

This work was supported by the National Natural Science Foundation of China (Grant No. 30971555 and 30671124), National Basic Research Program of China (Grant No.2012CB215300), National Transgenic Major Program of China (Grant Nos. 2008ZX003-002 and 2009ZX08009-093B).

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Correspondence to Jingjuan Yu.

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10725_2011_9651_MOESM1_ESM.ppt

Schematic diagram of Zm401 promoter. a sketch map illustrates the whole Zm401 promoter. b Nucleotide sequence of -670 to -510 of Zm401 promoter in detail. The DOFCORE was underlined. CAAT box: GGACCAATACT; DOFCORE: AAAG; LAT52 Q element: TGGTTA; NTP303 Q element: AAATGA; TATA box: TATTAAT (PPT 204 kb)

10725_2011_9651_MOESM2_ESM.ppt

Deduced amino acid sequences alignment among the cloned Zmdof1 gene and the published Zmdof1 (EMBL: CAA46875.1), Zmdof2 (EMBL:CAA56287.1), and Zmdof3 (EMBL:CAA56288.1) genes (Yanagisawa and Izui 1993). The four cysteine residues are indicated by arrows (PPT 195 kb)

10725_2011_9651_MOESM3_ESM.ppt

Two possible speculations that Zmdof1 suppresses Zm401. a Zmdof1 competes the Dof binding site with a Dof domain protein. b Zmdof1 works together with a (some) suppressing mediator to inhibit the expression of Zm401 (PPT 148 kb)

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Chen, X., Wang, D., Liu, C. et al. Maize transcription factor Zmdof1 involves in the regulation of Zm401 gene. Plant Growth Regul 66, 271–284 (2012). https://doi.org/10.1007/s10725-011-9651-5

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