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Isolation, optimization, and functional analysis of the cDNA encoding transcription factor OsDREB1B in Oryza Sativa L.

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Abstract

A previous study had indicated that the transcription factors DREB/CBF (DRE-binding protein/C-repeat binding factor) play important roles in the expression of many stress inducible genes under cold-temperature, dehydration and high-salt conditions. In this study, we have isolated a cDNA clone that encoded a DRE-binding protein from rice cDNA library using the yeast one-hybrid system with DRE cis-acting element in the promoter region of rd29A gene as bait. Sequence analysis of the deduced amino acid sequence showed this protein was a putative AP2/EREBP transcription factor with a conserved AP2/EREBP domain and a potential nuclear localization signal (NLS). Expression pattern studies of this DRE-binding protein revealed that this gene was not only strongly induced by cold-temperature as reported by previous study but also induced by high-temperature as well. For the purpose of analyzing this gene conveniently, we attempted to apply the codon optimization method to reconstruct the gene of transcription factor in plants. A new sequence having decreasing GC contents, secondary structures, optimized codons and identical amino acid sequence with native gene was synthesized, which named OsDREB1BI, and then this optimized gene was transformed into Arabidopsis thaliana cv. Columbia by floral dip method. Results indicated that the OsDREB1BI gene was over-expressed in transgenic plants under cold and high-temperature, meanwhile, those transgenic plants also revealed freezing and heat tolerance. These might lay a strong foundation for exploiting the freezing and heat tolerance of rice and other species.

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Acknowledgements

We thank Dr. Genlou Sun and Dr. Ken Kasha for critical reading of this manuscript. This work was supported by a grant from Jiangsu Natural foundation (BK2003042) and a part from IAEA (12228/RO), the Youth Fund of Shanghai Academy of Agricultural Sciences (2003–13), the National Special Program for Research and Industrialization of Transgenic plants (J00-A-007), the Shanghai Key Basic Research Project (03DJ14016, 03DZ1931) and natural science foundation of China (30370987).

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Authors and Affiliations

  1. College of Bioscience and Biotechnology, Yangzhou University, 88 Daxue Road, Yangzhou, 225009, The Peoples’s Republic of China

    Qiu-lin Qin & Jian-min Chen

  2. Agro-Biotechnology Research Center of Shanghai Academy of Agricultural Sciences, 2901 Beidi Road, Shanghai, 201106, The People’s Republic of China

    Qiu-lin Qin, Jin-ge Liu, Ri-he Peng, Ai-sheng Xiong & Quan-hong Yao

  3. Department of Horticulture, Nanjing Agricultural University, No. 1 Weigang, Nanjing, 210095, The People’s republic of China

    Jin-ge Liu & Zhen Zhang

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  1. Qiu-lin Qin
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  2. Jin-ge Liu
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  3. Zhen Zhang
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  4. Ri-he Peng
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  5. Ai-sheng Xiong
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  6. Quan-hong Yao
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  7. Jian-min Chen
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Correspondence to Quan-hong Yao or Jian-min Chen.

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The authors wish it to be known that, in their opinion, the first two authors should be regarded as joint First Authors

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Qin, Ql., Liu, Jg., Zhang, Z. et al. Isolation, optimization, and functional analysis of the cDNA encoding transcription factor OsDREB1B in Oryza Sativa L.. Mol Breeding 19, 329–340 (2007). https://doi.org/10.1007/s11032-006-9065-7

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  • DOI: https://doi.org/10.1007/s11032-006-9065-7

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