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Tobacco plants expressing the defensin NaD1 enhance drought tolerance characteristics in transgenic lines

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Abstract

Antimicrobial peptides (AMPs) play a pivotal role in plant immune responses to diverse stresses, and hence, have become the novel molecules for studying plant responses to environmental harsh conditions. For the first time, the effect of the antimicrobial peptide defensin NaD1 on drought stress was investigated in tobacco by generating the NaD1 transgenic lines overexpressing the coding sequence of NaD1. The NaD1 expression was confirmed by RT-PCR, and the NaD1 peptide presence was verified using ELISA and western blot analysis in the tobacco transgenic lines. In silico bioinformatic analysis revealed that the most abundant components in Cis-regulatory elements in NaD1 homologs in Nicotiana attenuate (NaDEF genes) are MYB, MYC and ABRE elements suggesting that the NaD1 promoter is involved in the abiotic stress regulation. The chlorophyll a, b, and total chlorophyll contents, and correspondingly, the index of chlorophyll stability (ICS) significantly (P ≤ 0.01) increased in all 3 NaD1 transgenic lines under drought stress. Moreover, activities of Catalase (CAT), Peroxidase (POD), Ascorbate peroxidase (APX) and Superoxide dismutase (SOD) were significantly enhanced in response to drought stress in the transgenic lines. Among the three transgenic lines, line 1 showed more tolerance to the drought stress. The data together suggest that the expression of NaD1 increases the antioxidant activity of the enzymes presumably leading to elimination of ROS levels and maintenance of the chlorophyll content and stability, resulting in enhanced drought tolerance in the transgenic tobacco lines. Therefore, the defensin NaD1 is an essential factor in regulation of plant responses to drought stress.

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  • 27 July 2023

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Acknowledgements

This research was carried out with partially financial assistance of Iran National Science Foundation (INSF) under number of 98022316. This study was financially supported by grant NO: biodc-13085-24100.1 of the Biotechnology Development Council of the Islamic Republic of Iran.

Funding

This research was partly funded by University of Guilan, this research was carried out with partially financial assistance of Iran National Science Foundation (INSF) under number of 98022316. This study was financially supported by grant NO: biodc-13085-24100.1 of the Biotechnology Development Council of the Islamic Republic of Iran.

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Authors and Affiliations

  1. Department of Agricultural Biotechnology, Faculty of Agricultural Sciences, University of Guilan, Rasht, Iran

    Sara Royan & Reza Shirzadian-Khorramabad

  2. Department of Plant Protection, Faculty of Agricultural Sciences, University of Guilan, Rasht, Iran

    Arash Zibaee

  3. Department of Biology, Faculty of Science, Golestan University, Gorgan, Iran

    Mohammad B. Bagherieh-Najjar

  4. Production Engineering and Plant Genetic Department, Faculty of Agriculture and Natural Resources, Lorestan University, Khorramabad, Iran

    Farhad Nazarian Firouzabadi

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  1. Sara Royan
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SR did the research experiments, data analysis and wrote the first draft of manuscript. RSH conducted and supervised the whole research project and revised the manuscript. AZ, MBN and FNF cooperate as advisors in different parts of the work, respectively. All authors have read and approved the manuscript.

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Correspondence to Reza Shirzadian-Khorramabad.

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Communicated by Neelam sangwan.

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Royan, S., Shirzadian-Khorramabad, R., Zibaee, A. et al. Tobacco plants expressing the defensin NaD1 enhance drought tolerance characteristics in transgenic lines. Plant Growth Regul 101, 503–518 (2023). https://doi.org/10.1007/s10725-023-01037-6

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