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Cloning and Functional Analysis of the Promoter of a Stress-inducible Gene (ZmRXO1) in Maize

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Abstract

The ZmRXO1 gene is a nucleotide-binding site leucine-rich repeat (NBS–LRR) type of R gene in maize (Zea mays). To understand the regulatory mechanism of ZmRXO1 gene expression, we isolated and characterized the ZmRXO1 promoter (PZmRXO1)—the 5′ flanking region of ZmRXO1. A series of PZmRXO1 deletion derivatives, R1–R4, from the translation start code (−1,576, −934, −829, and −582) were fused to the GUS reporter gene, and each deletion construct was analyzed by Agrobacterium-mediated transformation into tobacco. Sequence analysis showed that several cis-acting elements (MBS, Box-I, TGA-element and CCAAT-box) were located within the promoter. Deletion analysis of the promoter suggested that the 1,576-bp fragment upstream of ZmRXO1 gene showed a high level of GUS expression in tobacco. The promoter sequence (−582 to −1) was sufficient to improve transcription of GUS gene under hormones (MeJA, GA, ABA), drought and low temperature. Moreover, there might be repressor elements in the region (−1,576 to −934 bp) to repress ZmRXO1 gene expression under treatment with salicylic acid.

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Acknowledgments

This research was funded by the Twelfth Five Year Plan Project of Science and Technology Support, P.R. China (2012BAD19B04, 2014BAD14B02), the Ministry of Agriculture Key Project of GM Cultivation of New Varieties, P. R. China (2013ZX08004004), the Project of International Collaboration Plan in Jilin Province (20100723) and the Research and Development of Industrial Technology Special at Jilin Provincial Development and Reform Commission (2013C001).

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Correspondence to Hongyu Pan.

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Ye Tao and Fengting Wang contributed equally to this work and are co-first authors.

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Tao, Y., Wang, F., Jia, D. et al. Cloning and Functional Analysis of the Promoter of a Stress-inducible Gene (ZmRXO1) in Maize. Plant Mol Biol Rep 33, 200–208 (2015). https://doi.org/10.1007/s11105-014-0741-1

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