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Isolation and Characterization of the Cold-Induced Phyllostachys edulis AP2/ERF Family Transcription Factor, peDREB1

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Abstract

The dehydration-responsive element binding (DREB) transcription factor binds to the dehydration-responsive element (DRE), a cis-acting element, and regulates the expression of multiple target genes involved in plant tolerance to drought, salinity, and cold stress. Phyllostachys edulis is one of the most widely cultivated bamboos species, which is adapted for cultivation in warm and wet condition, and is sensitive to cold. However, a DREB homolog gene has not yet been identified in P. edulis. In this paper, we report the characterization of the peDREB1, cloned from cold-induced P. edulis by rapid amplification of cDNA ends (RACE). The characterization of the gene product as a DREB transcription factor is supported the nuclear localization of a transiently expressed green fluorescent protein (GFP) fusion in onion epidermal cells. Furthermore, peDREB1 can activate reporter gene expression, and we show that the protein specifically binds to the conserved DRE element in a yeast one-hybrid assay. Expression analysis shows that peDREB1 transcription levels rapidly accumulate following exposure to cold stress, peaking at 3 h, while only small changes in mRNA expression are observed during abscisic acid, osmotic stress, or salt treatment. Thus, we have identified and characterized the peDREB1 transcription factor. Similar to other APETALA2/ethylene-responsive element-binding proteins (AP2/EREBP) regulators, we report that the peDREB1 homolog is activated in response to environmental factors in tropical plants.

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Abbreviations

ABA:

Abscisic acid

DRE:

Dehydration-responsive element

DREB:

Dehydration-responsive element-binding protein

RACE:

Rapid amplification of cDNA ends

NLS:

Nuclear localization signal

RT–PCR:

Semi-quantitative reverse transcription PCR

GFP:

Green fluorescent protein

AP2/EREBP:

APETALA2/ethylene-responsive element binding protein

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Acknowledgments

We thank Professor Shen Shihua (Institute of Plant Botany, Chinese Academy of Science, Beijing, P. R. China) for providing the wild-type and mutant (mDRE) strains of yeast and Professor Ma Youzhi (Chinese Academy of Agricultural Science, Beijing, P. R. China) for providing the vector 163GFP. This work was supported by grants from the Special Fund Project for the Scientific Research of the Forestry Public Welfare Industry (no. 200704001) and the Zhejiang Municipal Science and Technology Project (no. 2009 C12097).

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Correspondence to Lu-Bin Li or Cong-Fen He.

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Lei Liu and Xiao-Lu Cao contributed equally to this work.

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Liu, L., Cao, XL., Bai, R. et al. Isolation and Characterization of the Cold-Induced Phyllostachys edulis AP2/ERF Family Transcription Factor, peDREB1 . Plant Mol Biol Rep 30, 679–689 (2012). https://doi.org/10.1007/s11105-011-0380-8

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