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Heterologous expression of PDH47 confers drought tolerance in indica rice

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Abstract

In response to challenge of producing rice plants tolerant to drought stress, Pea DNA Helicase 47 (PDH47) from Pisum sativum under the control of constitutive 35S CaMV promotor was introduced into indica rice cultivar ASD16 mediated through Agrobacterium tumefaciens using immature embryos as explant. PCR positive transgenic rice lines showed varied levels of PDH47 transcripts as evident by semi-quantitative reverse transcription PCR (RT-PCR). Copy number of the transgene was determined through quantitative PCR (qPCR) analysis. T2 transgenic lines showed upregulation of PDH47 transcripts both in leaf and root tissues after 30 days of drought stress treatment in pots as evident by quantitative reverse transcription PCR (qRT-PCR). The upregulation of PDH47 transcripts during drought stress correlated with increased accumulation of osmolytes like proline, increased relative water content and decreased accumulation of hydrogen peroxide (H2O2). T2 transgenic lines also showed enhanced tolerance to drought compared to wild type rice after withdrawal of water for 30 days. Further, the heterologous expression of PDH47 regulated several endogenous stress-responsive genes in transgenic rice during drought stress. To the best of our knowledge, we are reporting for the first time the involvement of PDH47 transgene imparting drought tolerance in transgenic rice, demonstrating its potential role in crop improvement.

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Acknowledgements

This work was financially supported by Department of Biotechnology, Government of India. We gratefully acknowledge the Distributed Information Centre, Department of Agricultural Biotechnology, DBT-AAU Centre, Assam Agricultural University, Jorhat for providing necessary facilities to undertake this research work.

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Authors and Affiliations

  1. Department of Agricultural Biotechnology, Assam Agricultural University, Jorhat, Assam, 785013, India

    Dhanawantari L. Singha, Dimple Boro & Salvinder Singh

  2. Plant Molecular Biology Group, International Centre for Genetic Engineering and Biotechnology, Aruna Asaf Ali Marg, New Delhi, 110067, India

    Narendra Tuteja

  3. Director of Research (Agri), Assam Agricultural University, Jorhat, Assam, 785013, India

    Girindra Nath Hazarika

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  1. Dhanawantari L. Singha
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  5. Salvinder Singh
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DLS designed and performed the experiments and analyzed the data; DB collected the data; SS supervised the research; DLS wrote the manuscript with critical inputs from SS, GNH and NT.

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Correspondence to Salvinder Singh.

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11240_2017_1248_MOESM2_ESM.tif

Fig. S1 Agrobacterium tumefaciens-mediated genetic transformation of indica rice cv. ASD16 using immature embryos. A Co-cultivation of immature embryos with Agrobacterium strain AGL1 harbouring p1301-PDH47 for 7 days; B, C and D Immature embryos in first, second and third resting medium, respectively containing 250mg/L cefotaxime and 150mg/L timentin after 14 days of culture; E, F and G Immature embryos in first, second and third selection medium, respectively containing 30mg/L hygromycin B at 14 days interval; H Induction of green embryoids on CC regeneration medium after 7 days of culture; I Rooting of the in vitro regenerated shoots after 15 days of culture; J Putative transgenic lines maintained in the transgenic green house. (TIF 16983 KB)

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Singha, D.L., Tuteja, N., Boro, D. et al. Heterologous expression of PDH47 confers drought tolerance in indica rice. Plant Cell Tiss Organ Cult 130, 577–589 (2017). https://doi.org/10.1007/s11240-017-1248-x

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