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Lobularia maritima thioredoxin-h2 gene mitigates salt and osmotic stress damage in tobacco by modeling plant antioxidant system

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Abstract

Thioredoxins (Trxs) are multi-functional redox proteins characterized by a conserved redox-active CGPC site. In plants, Trxs are small antioxidant proteins encoded by a multigene family and regulate several processes during development and growth. Consequently, Thrxs are attractive candidates for transgenic plant improvement to overcome the abnormal plant growth and development observed during abiotic stress conditions. Here, we isolated the gene encoding the h-type Trx protein, LmTrxh2, from the halophyte plant Lobularia maritima and developed transgenic tobacco plants that overexpress LmTrxh2. Based on an in silico sequential, phylogenetic, and three-dimensional model analysis, LmTrxh2 protein shares high sequence identity and common structural characteristics with plant thioredoxins. RT-qPCR analysis revealed differential temporal and spatial regulation of LmTrxh2 in L. maritima exposed to salt (150 mM NaCl), osmotic (10% PEG 8000), and oxidative (10 µM H2O2) stresses. Subcellular localization using confocal microscopy revealed that LmTrxh2 localized to the plasma membrane. Transgenic LmTrxh2 tobacco lines exhibited improved salt and osmotic stress tolerance compared to non-transgenic (NT) plants, and this tolerance was associated with mitigation of oxidative damage. Finally, four stress-related genes were upregulated in LmTrxh2 transgenic tobacco plants compared to those in NT plants under control and stress conditions. These findings suggest that LmTrxh2 is an attractive candidate gene for the genetic engineering of crops with enhanced salt and osmotic stress tolerance.

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Acknowledgements

This work was funded by grants provided by the Tunisian Ministry of Higher Education and Scientific Research (Program contract 2019–2022).

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Authors and Affiliations

  1. Biotechnology and Plant Improvement Laboratory, Centre of Biotechnology of Sfax, University of Sfax, B.P 1177, 3078, Sfax, Tunisia

    Rania Ben Saad, Walid Ben Romdhane, Mohamed Taieb Bouteraa, Olfa Jrad & Anis Ben Hsouna

  2. Plant Production Department, College of Food and Agricultural Sciences, King Saud University, P.O. Box 2460, 11451, Riyadh, Saudi Arabia

    Walid Ben Romdhane

  3. Departments of Life Sciences, Faculty of Sciences of Gafsa, Zarroug, 2112, Gafsa, Tunisia

    Anis Ben Hsouna

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  1. Rania Ben Saad
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  5. Anis Ben Hsouna
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Contributions

RBS conceived the study conception and design. Experiments, data collection, and data analysis were performed by OJ, MTB, ABH, RBS and WBR. The manuscript was written by RBS and WBR. All authors read and approved the manuscript.

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Correspondence to Rania Ben Saad.

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Communicated by Neelam Sangwan.

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10725_2022_805_MOESM1_ESM.tif

Amino acid sequences alignment of LmTrxh2 and its homologs belonging to plant thioredoxins. The location of the thioredoxin superfamily conserved domain is surrounded by a green dashed line, and the position of thioredoxin motif is surrounded by a solid red box. The two catalytic conserved cysteine residues were labeled with blew Arrows. GenBank accession numbers of different plant thioredoxins proteins used in this multiple alignment are arranged as follows: A. thaliana (NP_198811), C. sativa (XP_019089899), A. lyrata (XP_020872997), C. rubella (XP_006284753), E. salsugineum (XP_024014606), B. oleracea (XP_013589887), R. sativus (XP_018446242), B. rapa (XP_009125122), T. hassleriana (XP_010531799), A. duranensis (XP_015962819), G. soja (KHN13657), T. aestivum (ACH61777), O. sativa (XP_015646978), and O. sativa (XP_015631704). Supplementary Material 1. 22660 kb

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Ben Saad, R., Ben Romdhane, W., Bouteraa, M.T. et al. Lobularia maritima thioredoxin-h2 gene mitigates salt and osmotic stress damage in tobacco by modeling plant antioxidant system. Plant Growth Regul 97, 101–115 (2022). https://doi.org/10.1007/s10725-022-00805-0

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