Ectopic Expression of Rice PYL3 Enhances Cold and Drought Tolerance in Arabidopsis thaliana
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Abscisic acid (ABA) plays an important role in plant development and adaptation to abiotic stresses. The pyrabactin resistance-like (PYL) gene family has been characterized as intracellular ABA receptors in Arabidopsis. We describe here the functional characterization of PYL3 ABA receptor from a drought-tolerant rice landrace Nagina 22 (N22). The induced expression level of the PYL3 transcript was observed in the N22 under different stress treatments, including cold, drought, high temperature, salt and ABA. In contrast, the expression of PYL3 was down-regulated in drought-susceptible rice cv. IR64 in response to above stresses. C-terminal GFP translational fusion of OsPYL3 was localized to both cytosol and nucleus explaining in part functional conservation of PYL protein as ABA receptor. Arabidopsis transgenic lines overexpressing OsPYL3 were hypersensitive to ABA suggesting ABA signaling pathway-dependent molecular response of the OsPYL3. Further, constitutive overexpression of OsPYL3 in Arabidopsis led to improved cold and drought stress tolerance. Thus, OsPYL3 identified in this study could be a good candidate for genetic improvement of cold and drought stress tolerance of rice and other crop plants.
KeywordsABA receptor Rice Cold Drought PYL Abiotic stress
This work was supported by the Indian Council of Agricultural Research (ICAR)-sponsored Network Project on Transgenics in Crops (NPTC). SKL acknowledges the University Grants Commission (UGC) and the Council of Scientific and Industrial Research (CSIR) for a CSIR-UGC JRF and SRF fellowship. SKM acknowledges the Department of Science and Technology (DST) for a DST-INSPIRE fellowship. The plants were grown in a space provided by the National Phytotron Facility, IARI. VC was supported by NASF (ICAR) project (Grant No. NFBSFARA/Phen 2015). Assistance provided by Mr. Amit K. Singh and Mr. Jeet B. Singh for growing rice and Arabidopsis plants is acknowledged.
SKL did all the experiments, experimental design and drafted the manuscript. SKM performed ABA and cold phenotyping of the transgenic Arabidopsis on plates. VC and KCB participated in experimental design and manuscript writing. All authors read and approved the final manuscript.
- 4.Bansal, K. C., Lenka, S. K., & Tuteja, N. (2011). Abscisic acid in abiotic stress tolerance: an ‘omics’ approach. In N. Tuteja, S. S. Gill, & R. Tuteja (Eds.), Omics and plant abiotic stress tolerance (pp. 143–150). Sharjah: Bentham Science.Google Scholar
- 5.Bello, B., Zhang, X., Liu, C., Yang, Z., Wang, Q., Zhao, G., et al. (2014). Cloning of Gossypium hirsutum sucrose non-fermenting 1-related protein kinase 2 gene (GhSnRK2) and its overexpression in transgenic Arabidopsis escalates drought and low temperature tolerance. PLoS ONE, 9(11), e112269. https://doi.org/10.1371/journal.pone.0112269.CrossRefGoogle Scholar
- 15.Gonzalez-Guzman, M., Pizzio, G. A., Antoni, R., Vera-Sirera, F., Merilo, E., Bassel, G. W., et al. (2012). Arabidopsis PYR/PYL/RCAR receptors play a major role in quantitative regulation of stomatal aperture and transcriptional response to abscisic acid. The Plant Cell, 24(6), 2483–2496.CrossRefGoogle Scholar
- 22.Kim, H., Hwang, H., Hong, J. W., Lee, Y. N., Ahn, I. P., Yoon, I. S., et al. (2012). A rice orthologue of the ABA receptor, OsPYL/RCAR5, is a positive regulator of the ABA signal transduction pathway in seed germination and early seedling growth. Journal of Experimental Botany, 63(2), 1013–1024. https://doi.org/10.1093/jxb/err338.CrossRefGoogle Scholar
- 25.Koornneef, M., Jorna, M. L., Brinkhorst-van der Swan, D. L., & Karssen, C. M. (1982). The isolation of abscisic acid (ABA) deficient mutants by selection of induced revertants in non-germinating gibberellin sensitive lines of Arabidopsis thaliana (L.) heynh. Theoretical and Applied Genetics, 61(4), 385–393. https://doi.org/10.1007/BF00272861.Google Scholar
- 26.Lenka, S. K. (2010). Characterization of Abiotic stress responsive cis-elements and transcription factor genes from rice (Oryza sativa L.). Dissertation, Indian Agricultural Research Institute, New Delhi.Google Scholar
- 27.Lenka, S. K., Katiyar, A., Chinnusamy, V., & Bansal, K. C. (2010). Comparative analysis of drought-responsive transcriptome in Indica rice genotypes with contrasting drought tolerance. Plant Biotechnology Journal, 9(3), 315–327. https://doi.org/10.1111/j.1467-7652.2010.00560.x.CrossRefGoogle Scholar
- 28.Lenka, S. K., Lohia, B., Kumar, A., Chinnusamy, V., & Bansal, K. C. (2009). Genome-wide targeted prediction of ABA responsive genes in rice based on over-represented cis-motif in co-expressed genes. Plant Molecular Biology, 69(3), 261–271. https://doi.org/10.1007/s11103-008-9423-4.CrossRefGoogle Scholar
- 29.Lenka, S. K., Nims, N. E., Vongpaseuth, K., Boshar, R. A., Roberts, S. C., & Walker, E. L. (2015). Jasmonate-responsive expression of paclitaxel biosynthesis genes in Taxus cuspidata cultured cells is negatively regulated by the bHLH transcription factors TcJAMYC1, TcJAMYC2, and TcJAMYC4. Frontiers in Plant Science, 6, 115. https://doi.org/10.3389/fpls.2015.00115.CrossRefGoogle Scholar
- 31.Marin, E., Nussaume, L., Quesada, A., Gonneau, M., Sotta, B., Hugueney, P., et al. (1996). Molecular identification of zeaxanthin epoxidase of Nicotiana plumbaginifolia, a gene involved in abscisic acid biosynthesis and corresponding to the ABA locus of Arabidopsis thaliana. EMBO Journal, 15(10), 2331–2342.Google Scholar
- 33.Nambara, E., & Marion-Poll, A. (2005). Abscisic acid biosynthesis and catabolism. Annual Review of Plant Biology, 56, 165–185. https://doi.org/10.1146/annurev.arplant.56.032604.144046.CrossRefGoogle Scholar
- 37.Pizzio, G. A., Rodriguez, L., Antoni, R., Gonzalez-Guzman, M., Yunta, C., Merilo, E., et al. (2013). The PYL4 A194T mutant uncovers a key role of PYR1-LIKE4/PROTEIN PHOSPHATASE 2CA interaction for abscisic acid signaling and plant drought resistance. Plant Physiology, 163(1), 441–455. https://doi.org/10.1104/pp.113.224162.CrossRefGoogle Scholar
- 48.Yu, J., Yang, L., Liu, X., Tang, R., Wang, Y., Ge, H., et al. (2016). Overexpression of poplar Pyrabactin resistance-like abscisic acid receptors promotes abscisic acid sensitivity and drought resistance in transgenic Arabidopsis. PLoS ONE, 11(12), e0168040. https://doi.org/10.1371/journal.pone.0168040.CrossRefGoogle Scholar