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Functional Characterization of a Chrysanthemum dichrum Stress-Related Promoter

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Abstract

A 1,474-bp stress-inducible CdDREBa promoter was identified from Chrysanthemum dichrum, revealing several candidate stress-related cis-acting elements (MYC-box, MYB site, GT-1, and W-box) within it. In Arabidopsis leaf tissues transformed with a CdDREBa promoter-β-glucuronidase (GUS) gene fusion, serially 5′-deleted CdDREBa promoters were differentially activated by cold and salinity. Histochemical and quantitative assays of GUS expression allowed us to localize a critical part of the promoter located between upstream 430 and 351 nt. This 80-bp fragment enhanced GUS expression under salinity stress when fused to −90/+8 CaMV 35S minimal promoter. Further promoter internal-deletion assays indicated that a low temperature-responsive element was located between positions −430 and −390, and a salinity inducible one between −385 and −351. Our results showed that there was a novel stress-related critical region except for the known cis-acting element (MYC-box, GT-1) in CdDREBa promoter.

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Abbreviations

GUS:

β-Glucuronidase

DREB/CBF:

Dehydration-responsive element binding/C-repeat binding factor

TAIL-PCR:

Thermal asymmetric interlaced PCR

M-MLV:

Maloney marine leukemia virus

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Acknowledgments

This study was supported by the Program for Hi-Tech Research, Jiangsu, China (Grant No. BE2008302, BE2009317), and the National Natural Science Foundation of China (Grant No. 30872064, 31071820, 31071825), and the Fundamental Research Funds for the Central Universities (KYJ200907, KYZ201112). The Program for New Century Excellent Talents in University of Chinese Ministry of Education (Grant No. NCET-10-0492).

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  1. College of Horticulture, Nanjing Agricultural University, Nanjing, Jiangsu, 210095, People’s Republic of China

    Yu Chen, Sumei Chen, Fadi Chen, Pirui Li, Lin Chen, Zhiyong Guan & Qingshan Chang

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  1. Yu Chen
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Correspondence to Fadi Chen.

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Chen, Y., Chen, S., Chen, F. et al. Functional Characterization of a Chrysanthemum dichrum Stress-Related Promoter. Mol Biotechnol 52, 161–169 (2012). https://doi.org/10.1007/s12033-011-9483-6

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