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Improved drought and salt stress tolerance in transgenic tobacco overexpressing a novel A20/AN1 zinc-finger “AlSAP” gene isolated from the halophyte grass Aeluropus littoralis

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Abstract

We describe here the isolation of a novel gene, designated AlSAP, from A. littoralis in a first step to exploit the potential of this halophyte grass as a genetic resource to improve salt and drought tolerance in plants and, particularly, in cereals. The Aeluropus genome contains a single AlSAP gene which has an intron at its 5’UTR. Sequence homology analysis showed that the AlSAP protein is characterized by the presence of two conserved zinc-finger domains A20 and AN1. AlSAP is induced not only by various abiotic stresses such as salt, osmotic, heat and cold but, also by abscisic acid (ABA) and salicylic acid (SA). Tobacco plants expressing the AlSAP gene under the control of the duplicated CaMV35S promoter exhibited an enhanced tolerance to abiotic stresses such as salinity (350 mM NaCl), drought (soil Relative Water Content (RWC) = 25%), heat (55°C for 2.5 h) and freezing (−20°C for 3 h). Moreover, under high salt and drought conditions, the transgenic plants were able to complete their life cycle and to produce viable seeds while the wild-type plants died at the vegetative stage. Measurements of the leaf RWC and of the root and leaf endogenous Na+ and K+ levels in AlSAP transgenic lines compared to wild-type tobacco, showed an evident lower water loss rate and a higher Na+ accumulation in senescent-basal leaves, respectively. Finally, we found that the steady state levels of transcripts of eight stress-related genes were higher in AlSAP transgenic lines than in wild-type tobacco. Taken together, these results show that AlSAP is a potentially useful candidate gene for engineering drought and salt tolerance in cultivated plants.

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Acknowledgments

Special thanks are extended to C. Périn and M. Conte (Plant Development and Genetic Improvement (DAP) unit of CIRAD) as well as to Kh. Belhaj (CBS), A. Price (University of Aberdeen, UK) for their critical review of the manuscript. The authors are also grateful to S. Abid, a teacher of English, for the English revision. Part of this work was conducted under the REFUGE platform funded by Agropolis Fondation, Montpellier France. This study was supported by a grant from Ministry of Higher Education Scientific Research and Technology of Tunisia (contrat programme 2006–2010) and by the European project CEDROME (INCO-CT-2005-015468).

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Authors and Affiliations

  1. Centre of Biotechnology of Sfax (CBS), BP”1117”, 3018, Sfax, Tunisia

    Rania Ben Saad, Nabil Zouari, Walid Ben Ramdhan, Jalel Azaza & Afif Hassairi

  2. CIRAD, UMR DAP, Avenue Agropolis, 34398, Montpellier, Cedex5, France

    Donaldo Meynard & Emmanuel Guiderdoni

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  1. Rania Ben Saad
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  2. Nabil Zouari
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  3. Walid Ben Ramdhan
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  4. Jalel Azaza
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  7. Afif Hassairi
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Correspondence to Afif Hassairi.

Additional information

R. Ben Saad and A. Hassairi should be considered as first authors, as they contributed equally to this work.

The AlSAP sequence was deposited to the Genbank with the accession number DQ0885218 (UniProtKB: A1YAQ3).

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11103_2009_9560_MOESM1_ESM.tif

Supplementary Fig. 1 The nucleotide (707 bp) and the deduced amino acid sequence (159 aa) of AlSAP gene and its genomic structure. (a)The position of 5’UTR (118 bp) and 3’UTR (112 bp) are marked by line. The position of start (atg) and the stop (tga) codons are indicated by *. The A20 and AN1 domains of the peptide are indicated by dark and light grey shading respectively. The predicted serine (S), threonine (T) and tyrosine (Y) phosphorylation sites by NetPhos 2.0 Server (http://www.cbs.dtu.dk/services/NetPhos/) with a score > 0.8 are indicated by a circle. The predicted of the threonine kinase specific protein phosphoylation site (score 0.9) by NetPhosK 1.0 Server is indicated by a square. (b) The nucleotide sequence of AlSAP gene showing the presence of an intron (italic letters, 1,523 bp) in the 5’UTR (light grey shading), the ORF and the 3’UTR (dark grey shading) (TIFF 800 kb)

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Ben Saad, R., Zouari, N., Ben Ramdhan, W. et al. Improved drought and salt stress tolerance in transgenic tobacco overexpressing a novel A20/AN1 zinc-finger “AlSAP” gene isolated from the halophyte grass Aeluropus littoralis . Plant Mol Biol 72, 171–190 (2010). https://doi.org/10.1007/s11103-009-9560-4

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