Abstract
Publicly available microarray data and RNA gel blot analysis identified a rice WRKY transcription factor, OsWRKY71, that is highly upregulated in response to cold stress. Experiments with OsWRKY71 promoter:GFP transgenic rice confirmed its cold-inducible expression. Transient expression of OsWRKY71-GFP in maize mesophyll protoplasts indicated that it is localized predominantly in the nucleus and to a lesser extent in the cytosol. Transcriptional activation assays revealed that OsWRKY71 suppresses luciferase reporter activity in maize protoplasts, suggesting that it functions as a transcriptional repressor in rice. To characterize the function of OsWRKY71 in rice, we generated transgenic rice plants carrying CaMV35S promoter:OsWRKY71. Upon cold (4 °C) treatment, two selected OsWRKY71 transgenic lines, OX12 and OX21, recovered much better with respect to survival rate, photosynthetic ability, fresh weight, and dry weight than the control lines. RT-PCR analysis of known cold-responsive genes found that expression of OsTGFR and WSI76 was increased in OsWRKY71 transgenic lines in response to cold stress. Our results suggest that OsWRKY71 has a positive function in cold tolerance by regulating downstream target genes.
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Acknowledgments
This work was supported by grants from the Mid-Career Researcher Program (NRF-2013R1A2A2A01068887 to J.-S. Jeon) and Basic Science Research Program (NRF-2014R1A1A1003903 to S.-K. Lee) of the National Research Foundation.
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C.-Y. Kim, K. T. X. Vo, and C. D. Nguyen have contributed equally to this work.
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Kim, CY., Vo, K.T.X., Nguyen, C.D. et al. Functional analysis of a cold-responsive rice WRKY gene, OsWRKY71 . Plant Biotechnol Rep 10, 13–23 (2016). https://doi.org/10.1007/s11816-015-0383-2
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DOI: https://doi.org/10.1007/s11816-015-0383-2