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Expression analysis of proteinase inhibitor-II under OsRGLP2 promoter in response to wounding and signaling molecules in transgenic Nicotiana benthamiana

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Abstract

Proteinase inhibitor-II (PI-II) genes are important defense related genes that play critical regulatory roles in plant growth and development. In the present study, the expression of tomato PI-II gene was investigated under the control of a wound-inducible OsRGLP2 (Oryza sativa root germin like protein 2) promoter in transgenic tobacco plants after wounding, ABA and MeJA applications. Transcript level of target gene in transgenic plants was confirmed by quantitative real time PCR (qPCR). In response to ABA treatment at different concentrations, PI-II gene was strongly induced under OsRGLP2 promoter at higher concentration (100 μM), while considerable level of target gene expression was observed with MeJA application at 50 μM concentration. Upon wounding, relatively high PI-II gene expression was observed after 36-h treatment. Correspondingly, high GUS activity was detected at 36 h with histochemical assay and microscopic analysis in the vascular regions of leaves, stem and roots in wounded transgenic plants. This inducibility of PI-II gene by wounding, ABA and MeJA indirectly indicates its role in plant defense mechanism against biotic and abiotic stresses. Moreover, it was also suggested that ABA and MeJA dependent signaling pathways are involved in stimulation of PI-II gene. To the best of our knowledge, this is the first report describing the induction of PI-II gene under the regulation of OsRGLP2 promoter under stress conditions. The results of present research are useful for potential role of PI-II gene to improve stress tolerance in transgenic crops. Thus, efficacy of this gene can potentially be exploited to test the responses of different plants to various environmental stresses.

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Acknowledgements

This work was financially supported by the Higher Education Commission, Islamabad, Pakistan. We are grateful to the Department of Plant and Environmental Sciences, University of Copenhagen for providing research facilities. The authors are thankful to the Kirsten Jørgensen for her guidance to set up the qPCR.

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Authors and Affiliations

  1. Department of Plant Sciences, Quaid-i-Azam University, Islamabad, 45320, Pakistan

    Shazia Rehman, Ejaz Aziz & Tariq Mahmood

  2. Department of Plant and Environmental Sciences, Faculty of Sciences, University of Copenhagen, 1871 Frederiksberg C, Copenhagen, Denmark

    Bodil Jørgensen & Søren K. Rasmussen

  3. Department of Biotechnology, University of Azad Jammu and Kashmir, Muzaffarabad, Pakistan

    Wasim Akhtar

Authors
  1. Shazia Rehman
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  2. Bodil Jørgensen
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  3. Søren K. Rasmussen
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  4. Ejaz Aziz
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  6. Tariq Mahmood
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Correspondence to Tariq Mahmood.

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Rehman, S., Jørgensen, B., Rasmussen, S.K. et al. Expression analysis of proteinase inhibitor-II under OsRGLP2 promoter in response to wounding and signaling molecules in transgenic Nicotiana benthamiana. 3 Biotech 8, 51 (2018). https://doi.org/10.1007/s13205-017-1070-5

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