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Functional characterization of wheat myo-inositol oxygenase promoter under different abiotic stress conditions in Arabidopsis thaliana

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Abstract

The production of wheat is severely affected by abiotic stresses such as cold, drought, salinity, and high temperature. Although constitutive promoters are frequently used to regulate the expression of alien genes, these may lead to undesirable side-effects in transgenic plants. Therefore, identification and characterization of an inducible promoter that can express transgene only when exposed to stresses are of great importance in the genetic engineering of crop plants. Previous studies have indicated the abiotic stress-responsive behavior of myo-inositol oxygenase (MIOX) gene in different plants. Here, we isolated the MIOX gene promoter from wheat (TaMIOX). The in-silico analysis revealed the presence of various abiotic stress-responsive cis-elements in the promoter region. The TaMIOX promoter was fused with the UidA reporter gene and transformed into Arabidopsis thaliana. The T3 single-copy homozygous lines were analyzed for GUS activity using histochemical and fluorometric assays. Transcript expression of TaMIOX::UidA was significantly up-regulated by heat (five fold), cold (seven fold), and drought (five fold) stresses as compared to transgenic plants grown without stress-induced conditions. The CaMV35S::UidA plants showed very high GUS activity even in normal conditions. In contrast, the TaMIOX::UidA plants showed prominent GUS activity only in stress treatments (cold, heat, and drought), which suggests the inducible behavior of the TaMIOX promoter. The substrate myo-inositol feeding assay of TaMIOX::UidA plants showed lesser GUS activity as compared to plants treated in abiotic stress conditions. Results support that the TaMIOX promoter could be used as a potential candidate for conditional expression of the transgene in abiotic stress conditions.

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Abbreviations

PCR :

Polymerase chain reaction

TaMIOX :

Wheat myo-inositol oxygenase gene

UidA :

β-glucuronidase gene

NptII :

Neomycin phosphotransferase gene

MS:

Murashige and Skoog medium

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Acknowledgements

The authors are grateful to the National Agri-Food Biotechnology Institute (NABI), Department of Biotechnology (DBT), Council of Scientific and Industrial Research (CSIR), and Panjab University, Chandigarh for fellowship, research support and funds. Authors would like to acknowledge DBT-eLibrary Consortium (Del-CON) for providing access to online journals. Authors are thankful to Dr Margaret Avery, University of the Witwatersrand for useful suggestions to impove manuscript.

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Authors and Affiliations

  1. National Agri-Food Biotechnology Institute (NABI), Department of Biotechnology, Ministry of Science and Technology (Government of India), Sector-81, Knowledge City, S.A.S. Nagar, Mohali, 140306, India

    Anshu Alok & Siddharth Tiwari

  2. University Institute of Engineering and Technology, Panjab University, Chandigarh, 160014, India

    Anshu Alok & Jaspreet Kaur

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  1. Anshu Alok
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  2. Jaspreet Kaur
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Correspondence to Siddharth Tiwari.

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Alok, A., Kaur, J. & Tiwari, S. Functional characterization of wheat myo-inositol oxygenase promoter under different abiotic stress conditions in Arabidopsis thaliana. Biotechnol Lett 42, 2035–2047 (2020). https://doi.org/10.1007/s10529-020-02967-1

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