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Acta Agron Sin ›› 2011, Vol. 37 ›› Issue (08): 1389-1397.doi: 10.3724/SP.J.1006.2011.01389

• CROP GENETICS & BREEDING · GERMPLASM RESOURCES · MOLECULAR GENETICS • Previous Articles     Next Articles

TaCRF2 Gene Isolation from Triticum aestivum and Primary Function Validation in Tobacco (Nicotiana tabacum L.)

GUO Li-Xiang1,2,GAO Shi-Qing2,**,TANG Yi-Miao2,WANG Yong-Bo2,LIU Mei-Ying3,ZHANG Zhao3,XU Bei1,2,LIAN Wei-Wei1,2,ZHAO Chang-Ping2,*   

  1. 1 College of Life Sciences, Capital Normal University, Beijing 100048, China; 2 Beijing Hybrid Wheat Engineering and Technology Research Center, Beijing Academy of Agriculture and Forestry Sciences, Beijing 100097, China; 3 Department of Chemistry, Graduate School of University of Science and Technology of China, Hefei 230026, China
  • Received:2010-12-11 Revised:2011-03-06 Online:2011-08-12 Published:2011-05-11
  • Contact: 赵昌平, E-mail: zhaochangping401@vip.sohu.com, Tel: 13601096398

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Abstract: Wheat, like other crops, suffers from drought, salt, low-temperature and other abiotic stresses, seriously resulting in yield decline. It is an effective way for crop breeding to improve resistance via molecular biology techniques. Zinc finger protein, an important transcription factor commonly found in plant, can regulate the expression of multiple stress-inducible genes and enhance comprehensive resistances effectively. In this study, a RING-H2 zinc ?nger protein gene, designated as TaCRF2, was isolated from Triticum aestivum by RT-PCR. Its cDNA was 847 bp and encoded a putative protein of 280 amino acids with a predicted molecular mass of 30.97 kD and an isoelectric point (PI) of 7.03. A typical RING-H2 finger domain was found at the C-terminal region of TaCRF2 protein, and two transmembrane domains were found at the N-terminal region. Alignment of amino acid sequence showed that TaCRF2 was 82% identical to Oryza sativa putative RING zinc finger protein ABF95226. Subcellular localization analysis showed that the TaCRF2 was expressed in both nuclear and cytoplasm membrane. Real-time PCR showed that the transcript of TaCRF2 was strongly induced by drought, salinity and cold to some extent. The TaCRF2 gene was transformed into tobacco cultivar Wisconsin 38 (W38) by Agrobacterium mediated under the control of the CaMV 35S promoter. Under drought, salt and low-temperature stresses, transgenic tobacco lines carrying TaCRF2 gene developed the strong primary root, more lateral root and smaller yellow leaves, and performed higher tolerance to these stresses than the wide-type tobaccos, The primary function verification showed that overexpression of TaCRF2 gene in tobaccos enhanced their tolerance to drought and salinity stresses. This study will help further research of wheat RING-H2 type zinc finger protein reactions in the role of stress resistance, and bring wheat resistance breeding excellent candidate genes.

Key words: Tobacco, Triticum aestivum, RING-H2 zinc-finger protein, TaCRF2 gene, Drought stress, Salinity stress

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