Functional characterization of wheat myo -inositol oxygenase promoter under different abiotic stress conditions in Arabi
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ORIGINAL RESEARCH PAPER
Functional characterization of wheat myo-inositol oxygenase promoter under different abiotic stress conditions in Arabidopsis thaliana Anshu Alok . Jaspreet Kaur . Siddharth Tiwari
Received: 9 December 2019 / Accepted: 12 July 2020 Ó Springer Nature B.V. 2020
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 sideeffects 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 insilico 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
A. Alok S. Tiwari (&) 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 e-mail: [email protected]; [email protected]; [email protected]
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. Keywords Abiotic stress Arabidopsis thaliana Inducible promoter myo-inositol MIOX Wheat Abbreviations PCR Polymerase chain reaction TaMIOX Wheat myo-inositol oxygenase gene UidA b-glucuronidase gene NptII Neomycin phosphotransferase gene MS Murashige and Skoog medium
A. Alok J. Kaur University Institute of Engineering and Technology, Panjab University, Chandigarh 160014, India
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Biotechnol Lett
Introduction Wheat (Triticum aestivum L.) is a major food and feed crop for humans and animals, respectively, across the world. In India, 99.7 million tonnes of wheat was harvest from an area of over 29.58 million hectare
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