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Ectopic Expression of Rice PYL3 Enhances Cold and Drought Tolerance in Arabidopsis thaliana

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Abstract

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.

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Acknowledgements

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.

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Authors and Affiliations

  1. ICAR-National Research Centre on Plant Biotechnology, Indian Agricultural Research Institute, New Delhi, 110012, India

    Sangram K. Lenka, Senthilkumar K. Muthusamy & Kailash C. Bansal

  2. ICAR-Central Tuber Crops Research Institute, Thiruvananthapuram, 695017, India

    Senthilkumar K. Muthusamy

  3. Division of Plant Physiology, ICAR-Indian Agricultural Research Institute, New Delhi, 110012, India

    Viswanathan Chinnusamy

  4. TERI-Deakin NanoBiotechnology Centre, The Energy and Resources Institute, New Delhi, 110003, India

    Sangram K. Lenka & Kailash C. Bansal

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  1. Sangram K. Lenka
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  2. Senthilkumar K. Muthusamy
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  3. Viswanathan Chinnusamy
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  4. Kailash C. Bansal
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Contributions

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.

Corresponding author

Correspondence to Kailash C. Bansal.

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Supplemental Fig.

 1 Representative data showing screening of T2 OsPYL3 transgenic lines by RT-PCR. a.) Total RNA isolated from T2 plants b.) Expression of Tubulin (AtTub6) as a reference gene c.) Expression of randomly chosen OsPYL3 plants (T2) showing transgene expression. T2 plants showing OsPYL3 qualitative expression level equivalent to lane 6-8 are taken forward for further physiological analysis. (JPEG 81 kb)

Supplemental Fig.

 2 Quantitative estimation of root length and shoot biomass of WT and CaMV35S:OsPYL3 transgenic plants grown in 20 µM ABA compared to control. a.) Root length (measured in cm) of OsPYL3 overexpressing seedling was significantly (p value < 0.001) reduced in the ABA-containing medium compared to WT. b.) Shoot biomass (measured in g) of OsPYL3 transgenics was significantly lower (p value < 0.001) in ABA-containing medium compared to WT. (TIFF 24 kb)

Supplemental Fig.

 3 Phenotypic evaluation of OsPYL3 overexpressing lines (T2) grown in pots under cold and drought stresses. a). Cold stress at 4° C was imposed on both WT and OsPYL3 plants for 14 days after transplanting, b). Drought stress was imposed starting from the 14th day after transplanting till next 15 days by withholding water. (TIFF 619 kb)

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Lenka, S.K., Muthusamy, S.K., Chinnusamy, V. et al. Ectopic Expression of Rice PYL3 Enhances Cold and Drought Tolerance in Arabidopsis thaliana. Mol Biotechnol 60, 350–361 (2018). https://doi.org/10.1007/s12033-018-0076-5

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