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A TFIIIA-type zinc finger protein confers multiple abiotic stress tolerances in transgenic rice (Oryza sativa L.)

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Abstract

The TFIIIA-type zinc finger transcription factors are involved in plant development and abiotic stress responses. Most TFIIIA-type zinc finger proteins are transcription repressors due to existence of an EAR-motif in their amino acid sequences. In this work, we found that ZFP182, a TFIIIA-type zinc finger protein, forms a homodimer in the nucleus and exhibits trans-activation activity in yeast cells. The deletion analysis indicated that a Leu-rich region at C-terminus is required for the trans-activation. Overexpression of ZFP182 significantly enhanced multiple abiotic stress tolerances, including salt, cold and drought tolerances in transgenic rice. Overexpression of ZFP182 promotes accumulation of compatible osmolytes, such as free proline and soluble sugars, in transgenic rice. ZFP182 activates the expression of OsP5CS encoding pyrroline-5-carboxylate synthetase and OsLEA3 under stress conditions, while OsDREB1A and OsDREB1B were regulated by ZFP182 under both normal and stress conditions. Interestingly, site-directed mutagenesis assay showed that DRE-like elements in ZFP182 promoter are involved in dehydration-induced expression of ZFP182. The yeast two-hybrid assay revealed that ZFP182 interacted with several ribosomal proteins including ZIURP1, an ubiquitin fused to ribosomal protein L40. The in vivo and in vitro interactions of ZFP182 and ZIURP1 were further confirmed by bimolecular fluorescence complementation and His pull-down assays. Our studies provide new clues in the understanding of the mechanisms for TFIIIA-type zinc finger transcription factor mediated stress tolerance and a candidate gene for improving stress tolerance in crops.

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Acknowledgments

This work was supported by the Program for New Century Excellent Talents in University (No.NCET-08-0795), National Natural Science Foundation of China (Nos. 30971556, 31071069), State Key Laboratory of Rice Biology (No. 110101), the 111 project and the Fundamental Research Funds for the Central Universities (No.KYZ201137).

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Authors and Affiliations

  1. State Key Laboratory of Crop Genetics and Germplasm Enhancement, College of Agriculture, Nanjing Agricultural University, Nanjing, 210095, China

    Ji Huang, Shujing Sun, Dongqing Xu, Hongxia Lan, Hui Sun, Zhoufei Wang, Yongmei Bao, Jianfei Wang, Haijuan Tang & Hongsheng Zhang

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  1. Ji Huang
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  2. Shujing Sun
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  3. Dongqing Xu
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  4. Hongxia Lan
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  10. Hongsheng Zhang
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Correspondence to Hongsheng Zhang.

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Ji Huang and Shujing Sun contributed equally to this work.

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11103_2012_9955_MOESM1_ESM.tif

Fig. S1 ZFP182 gene expression. (a) Tissue-specific gene expression of ZFP182. The expression data are based on Rice Oligo Array Database (http://ricearray.org). (b) The mRNA expression of ZFP182 in response to salt (200 mM NaCl), cold (4oC) or 20 % PEG6000 treatment. The Actin gene was used as the internal RT-PCR control. (c) Gene expression of ZFP182 in response to drought stress in rice at three developmental stages. The gene expression data was derived from GEO database under accession number GSE26280 (Wang et al. 2011) (TIFF 178 kb)

Fig. S2 Real-time PCR analysis of ZFP182 expression in transgenic rice (TIFF 24 kb)

11103_2012_9955_MOESM3_ESM.tif

Fig. S3 Expression analysis of ZFP182, OsDREB1A and OsDREB1B in rice seedlings subjected to 20 % PEG6000 by semi-quantitative RT-PCR (TIFF 107 kb)

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Huang, J., Sun, S., Xu, D. et al. A TFIIIA-type zinc finger protein confers multiple abiotic stress tolerances in transgenic rice (Oryza sativa L.). Plant Mol Biol 80, 337–350 (2012). https://doi.org/10.1007/s11103-012-9955-5

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